Abstract Book

14 a 17 julho 2015
CI CTA 2015
10th Iberian and 7th Iberoamerican Congress
on Environmental Contamination and Toxicology
Abstract Book
CICTA 2015
10th Iberian and 7th Iberoamerican Congress
on Environmental Contamination and Toxicology.
Abstract Book
Autor: n.d.
Coordenação cientifica: Sandra Mariza Monteiro e Ana Maria Coimbra
Editor: UTAD - Universidade de Trás-os-Montes e Alto Douro
Ano: 2015
Suporte: Eletrónico
Formato: PDF / PDF/A
CI CTA 2015
10th Iberian and 7th Iberoamerican Congress
on Environmental Contamination and Toxicology
ISBN: 978-989-704-210-2
Secretariado: 3Victórias - Viagens e Turismo - Vila Real
Design: Magda Barata, Ana Mil-Homens
julho 2015
Coordenação cientifica: Sandra Mariza Monteiro e Ana Maria Coimbra
INDEX
COMMITTEES
25
PROGRAMME 31
Opening conference
Environmental sustainability. Insights to the future. Viriato Soromenho Marques
‘What hinders a more effective science-policy interface? How science needs
to change in the age of global environment and climate crisis’
41
Environmental Pharmaceuticals in Latin America, are a relevant issue today?43
WEDNESDAY 15.07.2015
Keynote Session: Vitor Vasconcelos
Cyanobacteria: toxins and other secondary metabolites and their
biotechnological applications.
45
Keynote Session: Julian Blasco
Sediment toxicity tests: tools for environmental quality.
47
Keynote Session: Manu Soto
New highlights for soil health assessment using earthworms: in vitro
and molecular tools. 49
Keynote Session: Afonso Celso Dias Bainy
Use of transcriptomic tools to identify new molecular and biochemical
biomarkers in aquatic non-model organisms
51
THURSDAY 16.07.2015 Keynote Session: Dick Vethaak
Micro and nanoplastics as emerging contaminants in the aquatic
environment: what we do and don’t know. 53
5
FRIDAY 17.07.2015 Keynote Session: Carlos Barata
ORAL COMMUNICATIONS 63
Emerging toxic compounds. 55
Aquatic Toxicology Keynote Session: Vance L., Trudeau
Keynote Session: Roman Ashauer
Histopathological baseline levels and confounding factors in sole
for marine health status assessment
Effect models at the core of ecotoxicology: insights and implications. 59
How human wellbeing is afecting the aquatic environment
61
65
65
In vitro inhibition by organophosphorus and carbamate compounds of
gilthead seabream brain acetylcholinesterase and hepatic carboxylesterase 67
Usefulness of fish head-kidney leucocytes to assess the
immunotoxicological effects of contaminants 69
Effects of carcinogenic versus non-carcinogenic polycyclic aromatic
hydrocarbons (PAHs) and their mixtures: an integrated biomarker survey
71
Interaction effects between metals and PAHs in the aquatic environment:
an in vivo study on genotoxicity and DNA repair with the zebrafish model 73
Effects of personal care products (PCP) on the MXR mechanism of
zebrafish embryos
75
Influence of a seawater pulse on the structure of freshwater
invertebrate communities
77
Effects of multigeneration exposures of D. magna to environmentally
relevant concentrations of nickel
79
Antioxidant responses and bioaccumulation in clam Scrobicularia
plana exposed to arsenate
81
Influence of temperature on the toxicity of cadmium and
benzo(a)anthracene to planktonic microalgae
83
Do cationic and hydrophobic modifications influence the
ecotoxicity of hydroxyethyl cellulose polymers?
85
A “stock culture independent” duckweed microbiotest with Spirodela
polyrhiza: a practical and low cost alternative to Lemna bioassays87
7
Endocrine Disruption
89
Risk Assessment 115
Effects of the endocrine disruptor vinclozolin in the benthic organism
Wastewater reuse: a study of chloroform formation
115
Chironomus riparius (Diptera).89
Carbon capture and storage (ccs) strategy: a risk assessment overview
Developmental effects of a xenoestrogen and an aromatase-inhibitor
focused on marine bacteria
in zebrafish (Danio rerio) embryogenesis
91
Differences in the xenoestrogenic sensitivity of five teleost species
93
Gonadal development and biological recovery of zebrafish exposed to
Testing potential CO2 leakages on three marine microalgae: metal
speciation and biological effects
95
Evaluation of zebrafish gonad development after endocrine
disruptors’ exposures
Maturity and stability parameters in the quality assessment of composts,
sludges and other representative organic wastes intended to agricultural use 119
exposed to 17α-ethinylestradiol
17ß-estradiol (e2) and 17α-ethinylestradiol (ee2)
nano-biosurfactants.123
97
99
Ecotoxicological and biochemical effects of an herbicide and a metal
on zooplankton and phytoplankton estuarine and marine species
99
The influence of different microalgae diets on cell and tissue level
biomarkers in mussel digestive gland
121
Rational design for safer nanomaterials: a case study with
Determination of estuarine vulnerability to contamination for use in
ecological risk assessment
Biomarkers 117
101
Accumulation kinetics of copper and silver and assessment of the effects
exerted after dietary exposure in oysters Crassostrea gigas103
125
Phosphorus fractionation in sediments from a small-sized dam in a rural
mountainous catchment: case study in NE Portugal
127
Environment risk assessment and bioaccumulation of metals in the metal
contaminated basin
129
Global Changes 131
An ecological relevant approach to oil sands
131
Bioremediation 133
Sub-lethal effects of exposure to atrazine in gill cells of sea lamprey
downstream migrants
105
Identification of molecular biomarkers of exposure to sanitary
sewage in oyster Crassostrea brasiliana
Synergistic and beneficial effects of 2,4-epibrassinolide and progesterone for the
107
Biomarkers, histopathology and condition indices in sole (Solea
senegalensis) exposed to contaminated sediment
environmental remediation of progesterone by Solanum nigrum L. Plants
Effect of aluminum on ros content and antioxidant system in rye
109
(Secale cereale l.) leaves and roots
Is there endocrine disruption in male mugilids from the Tagus Estuary? 111
Bioremediation of different types of oil in estuarine and coastal
Background concentrations of polycyclic aromatic hydrocarbons metabolites
environments – the role of autochthonous microorganisms
in Portuguese firemen
133
135
137
113
9
Emerging Toxic Compounds 139
Caracterización de la fotodegradación de fármacos en sistemas acuáticos
mediante cromatografía líquida-espectrometría de masas
139
Environmental Chemistry Monitoring 161
Microplastics in coastal sediments from South Portugal
161
Polychlorinated biphenyls in molluscs, within basque estuaries (northern Spain):
Impact of regulated and emerging pollutants and microplastics in marine
relationships with hydrological characteristics and human pressures
ecosystems (IMPACTA project)
Multibiomarker assessment of cadmium-based quantum dots effects in
141
Concerning pharmaceuticals, what are the portuguese wastewater
treatment plants hotspots of contamination?
the marine mussel Mytilus galloprovincialis
165
Nanotoxicology 167
143
Removal of veterinary antibiotics in constructed wetlands microcosms
– response of microbial community
163
145
Oxidative stress and enzymatic alterations induced by multi-walled carbon
nantoubes (MWCNTs) funcionalized with polyethylene glycol (PEG) in
Microbial Ecotoxicology 147
167
Effects of multi-stressors on juveniles of the common goby (Pomatoschistus
Relative sensitivities of life history variables of freshwater zooplankton
(rotifers and cladocerans) to stress: a review
mouse liver tissue
147
microps): microplastics, nanoparticles and temperature
169
Effect of mixed toxic diets (Microcystis aeruginosa and Scenedesmus acutus) on the
Interaction of graphene nanomaterials with liver derived cell lines:
competition of two cladocerans (Daphnia cf. mendotae and Simocephal)149
internalization, cytotoxicity and potentiation of chemical effects
Solid lipid nanoparticles affect leaf litter microbial colonizers and enzy-
Mechanisms of action and toxicity of silver nanoparticles in model
matic activities in freshwater
151
aquatic and terrestrial organisms
173
153
Soil Pollution, Conservation and Restoration 175
171
Effect of caffeine on the life table demography of the rotifer Plationus
patulus and the cladoceran Moina macrocopa Phytoavailability of lead in soils from an abandoned trap-shooting range 175
Ecosystem Level Effects 155
A new tool for water quality assessment combining distinct levels of
Environmental Chemistry and Toxicology organization155
Assessment of trace elements pollution and its environmental risk in
Inter-annual variability of soft bottom macrobenthic community of the NW
freshwater sediments of the Alqueva Reservoir (Guadiana Basin)
177
Gulf of Mexico in relationship with the DWH oil spill
The coastal lagoons of Tabasco state, Mexico: endangered ecosystems
179
157
Free-living nematodes as bio-indicators of the DWH oil spill in the mexican
exclusive economic zone, NW Gulf of Mexico
177
Is metal contamination in estuarine environments endangering european
159
eel (Anguilla anguilla) in their first stage in continental waters?
181
Eutrophication, water quality and nutrient loading: blooms of green algae. 183
11
Composites alto desempeño y cero voc´s para componentes automotrices y
Effect of cadium sublethal exposure on cellular redox balance and
aeroespaciales, a partir de resina poliéster insaturado dispersable en agua.185
oxygen consumption in the axolotl Ambystoma mexicanum.
Aguas residuales como fuente de aporte de piretroides en sedimentos
The study of biomarkers of toxic effects of oil in the background species
superficiales y organismos de Bahía Todos Santos, Baja California, México
187
of animals from oil contaminated territories
217
219
Can biochemical tools in benthic macroinvertebrates predict ecological
Cell and Molecular Toxicology 189
water quality of rivers?
Cytotoxicity of titanium dioxide nanoparticles in fish cell lines
189
Gender-dependent variability in lysosomal responses and stress on
221
DNA damage and marine pollutants: mechanisms of toxicity, applications and
stress of mussel Mytilus galloprovincialis223
handicaps of surveying genotoxicity in vivo in non-model organisms
A study on the Iberian hare as bioindicator of environmental pollution
191
Salinity effect on transcription levels in oysters Crassostrea brasiliana exposed
to phenanthrene
193
Exposure to phenanthrene and depuration: changes on gene transcription, enzymatic activity and lipoperoxidation in gill of scallops Nodipecten nodosus195
Developmental impairment following chronic exposure to ketamine in
early zebrafish (Danio rerio) embryos
197
POSTER SESSIONS 199
Biomarkers
207
In vivo and in vitro effects of anticholinesterase agents on acetylcholinesterase activities of different rabbit tissues
207
Bitter orange tree leaves as biomarkers of contamination by emerging industrial
pollutants in Seville city (Spain)
209
Oxidative stress biomarkers in liver and kidney of yelow-legged gulls 211
The use of Solea senegalensis Kaup, 1858 as an alternative model to
vertebrate animal testing: effects of a UV-filter to marine fish embryos
227
Lactuca sativa l. as a suitable crop for aluminum toxicity studies
229
Sediment health assessment of three estuaries of the Bay of Biscay using
cell and tissue level biomarkers in the polychaeta Nereis diversicolor
231
Temperature compensation in oxidative stress and biotransformation
enzyme activities in Mytilus galloprovincialis exposed to cypermethrin. 233
Biomonitoring of chemical pollution in rivers using biomarkers in
invertebrates from 2005 to 2014: a review
235
Hepatic and gill biomarker responses in Solea spp. from the NW
mediterranean fishing grounds
237
Influence of temperature on the toxicity of microplastics and hg to
the Mytilus galloprovincialis239
Transcriptinal levels of the encoding sulfotransferase in the oyster Crassostrea
Organochlorine pesticides in fat tissue of wild board fram Galicia (NW
Spain): influence of age and gender
225
213
brasiliana exposed in situ at Itacorubi mangrove in Santa Catarina, Brasil
241
Investigation of b-esterases in the freshwater fish matrinxã (brycon
amazonicus) exposed to cypermethrin based-insecticide
215
13
Global Changes
243
effects between different polycyclic aromatic hydrocarbons (PAHs)
O impacto das alterações climáticas na viticultura europeia segundo
os novos cenários do IPCC
An application of fish gill histopathology to address the interaction
243
271
Assessing the seasonal variability of biotic integrity indices in a mediterranean
Biological responses on C. maenas caused by sediment properties and pH interactions
stream exposed to industrial sewage discharge: a multi-taxa approach
273
during CO2 leakage events from sub-seabed injection and storage
245
Effects of crude extract of cyanobacteria (Radiocystis fernandoi) on the kidney
Reclamation and quality of technosols in Bulgaria
247
of traíra (Hoplias malabaricus): oxidative damage and histophatology
275
Toxicological effects of agricultural products kraft 36ec® and score 250ec®
and its active ingredients (abamectin and difenoconazole) in Danio rerio249
Toxic effects of fluoride (Fˉ) on the survival and behavior of the freshwater
snail Physella acuta (Draparnaud, 1805)
251
Effects of temperature and emerging contaminants of high concern on
juveniles of the common goby (Pomatoschistus microps)276
Mercury bioaccumulation in piscivorous fishes of an Amazon reservoir, Brazil 281
Evaluation of a endocrine disruptor (Vinclozolin) in embryos and adults
Use of the brain european sea bass (Dicentrarchus labrax) cell line
of Physa acuta (Gastropoda pulmonata).253
DLB-1 for toxicological studies Respuesta de la actividad mitocondrial en esporas de cyathea costaricensis
Enantiospecific chronic effects of ibuprofen in the freshwater crustacean,
para la evaluación de la calidad del agua del Río Bobos, México
A. desmarestii.285
255
Evaluation of the toxic effects of phthalates on natural populations
Effects of mercury bioaccumulation in gills of wild fish (Liza aurata)
of Chironomus riparius (Diptera): implications for ecotoxicity studies. 257
assessed by 1h nmr metabolomics and oxidative stress endpoints
Evaluation of ecotoxicity of two solvents: glycerol derived solvent
Fish response to the elevated potassium level: the model of the mining
vs ionic liquid
259
261
289
es in a eutrophic lagoon system of the central Gulf of California, México
291
Evidencia de efectos subletales en daphnia magna expuesta a sucesivos
Zinc, copper and cadmium accumulation in liver, kidney and muscle
tissues of lithobates catesbeianus tadpoles
287
Annual cycle of the microzooplanktonic aloricate and loricate ciliates abundanc-
Seasonal variation of micronuclei frequency in common sole of the
basque coast (SE Bay of Biscay)
area contamination.
283
263
shock tóxicos de licor negro
293
Soil Pollution, Conservation And Restoration
295
Transcriptional level of genes involved in the neurotransmitter system of Dicentrarchrus labrax in response to chronic exposure to psychopharmaceuticals 265
PAHs in soils affected by forest fires in Serra do Caramulo (Central Portugal) 295
Atrazine increases mitochondrial glutathione peroxidase activity
of gills of sea lamprey juveniles during salt acclimation
267
Microplastics in fish species of three Portuguese estuaries
269
How does metal(loid) pollution affect soil enzymatic activities at metal(loid)
enriched mine tailings? The role of pioneer plants rhizospheres
297
15
Chemical characteristics and toxicity of soils from an abandoned Pb/Zn mine 299
sediments from a reservoir used for public water supply São Paulo, Brazil329
Effects of multiple environmental stressors (thermal stress and cadmium
Metal and PAH assessment of the coastline and estuaries in the
exposure) at different times and levels of biological complexity in E. fetida301
north of Portugal
331
Copper fractionation in high resolution sampled vineyard soils managed
The implementation of an abiotic typology for brazilian reservoirs
333
under heroic viticulture in Ribeira Sacra (NW Spain)
Global amounts of polychlorinated biphenyls (PCBs) in the Porto
303
Microbial loop structure and occurrence of heavy metal resistant bacteria
coastline and Douro River estuary
in sediments of Araça Bay (Brazil): influence of contaminants
Total amounts of polycyclic aromatic hydrocarbons (PAHs) in the
307
Oporto district
Movilización de cobre por salpicadura de la lluvia en suelos de
cultivo de vid
309
compounds in the Aveiro Lagoon
Cu-based fungicides, by DLS, AF4-MALS and SP-ICP-MS
Global amounts of pesticides and their toxic impacts in
Study of the competitive release of heavy metals (Cu, Zn, Ni, Cd and Pb)
the Mondego River estuary
in a copper mine soil treated with ground mussel shell
Evolução do perfil físico-químico dos sedimentos do estuário
313
do Rio Lima
Mercury distribution among soil aggregate size fractions in spodic
horizons of temperate forest podzols from Galicia (NW Spain)
315
sampled temperate forest podzols from Galicia (NW Spain)
317
Rio Minho
por la actividad antrópica (Ushuaia, Tierra del Fuego, Argentina)
Diseño de un índice regional de calidad del agua para embalses
basado en la lógica difusa
Evaluation of riparian vegetation and invasive species in Galician
river ecosystems
321
abandoned Murçós W-Sn mine area (NE Portugal)
323
Monitoring the impacts on the ecosystem integrity of Portelo stream
Exploring the Potential of the Zeolite in Contaminated Soil
325
(Douro basin, NE Portugal) after a large spill of mining wastes
Microbiological contamination of Sargassum sp from north coast of São
Sediment quality assessment related to benthic macrofauna in
Paulo state (Brazil) by resistant bacteria
São Francisco River (Minas Gerais, Brazil)
327
343
345
347
349
Natural radiation and geochemical data of the Vila Pouca de
Aguiar massif, northern Portugal
Contamination of waters, stream sediments, soils and plants from the
341
Caracterização físico-química dos sedimentos do estuário do
Contenido de mercurio en suelos y sedimentos de cuencas hídricas afectadas
319
339
Evolução do perfil físico-químico dos sedimentos do estuário
do Rio Cávado
Distribution and accumulation of total hg in high vertical resolution
337
Spatial and annual distribution of estrogenic endocrine
Particle size characterization of throughfall from vine leaves sprayed with
311
335
351
353
355
Distributions of nutrients and metals in a sediment core and in superficial
17
Estudios preliminares de evaluación de toxicidad del fenitrotión en
cies representative of freshwater phytoplankton and zooplankton
dorada (Sparus aurata)357
Assessment of the pharmaceuticals removal by different wastewater
Diversidad y dinámica planctónica (fitoplancton y rotíferos) y la producción
treatments383
primaria de un lago urbano hipertrófico de la Ciudad de México.
Potential of Constructed Wetlands for the removal of antibiotic resistant
359
Eutrophication due to sewage discharges causes blooms of Peridinium
381
bacteria from livestock wastewater
385
Cell and Molecular Toxicology
387
387
quinquecorne (Dinophyceae) off the coasts of the
Port of Veracruz, Mexico: A case
361
Emerging Toxic Compounds 363
Genotoxic effects of rocket propellant’s component on rodents
Ecotoxicity of diltiazem
363
Interference of estrogenic and androgenic inputs in the brown trout
Ecotoxicology study of a mixture of drugs
365
lipid metabolism – experimental biochemical and molecular studies
Secale sylvestre: a wild rye potentially useful for aluminum tolerance
Chronic effects in crustacean Daphnia magna after exposure to a
solution of carbamazepine under single catalytic (TIO2) ozonation. 367
study391
Real time PCR analysis of NMDA receptor expression in zebrafish
Physiological alterations on the marine microalga Tetraselmis suecica
exposed to the uv filter benzophenone-3
389
369
embryos exposed to ketamine
393
Effects of psychopharmaceuticals exposure on RNA/DNA ratios in fish
Short term exposure to ketamine and its oxidative stress in
species371
zebrafish development
395
Microbial Ecotoxicology
397
Polystyrene microplastic localization and distribution in the gills
Evidences of salt stress on basidiomycete and zygomycete fungi
397
and digestive gland of mussel Mytilus galloprovincialis375
Impacto del cadmio en la actividad nitrificante del bacterioplancton
Impact of microcystin contaminated irrigation water on the physiology and
de los canales de Xochimilco (México): estudio en mesocosmos.
399
Nanotoxicology
401
Effects of antidepressant fluoxetine in European sea bass
juvenile´s behavior
mineral content of carrot (Daucus carota) – implications in crop quality
373
377
Effect of ibuprofen and carbamazepine on cyclooxygenase (COX-2)
gill gene expression, lipids and reproductive hormones in
Effects of silver nanoparticles on different soil exoenzymes activities 401
temperature conditioned juvenile Solea senegalensis379
Ecotoxicological comparison between industrial waste-recovered and
Influence of temperature on the toxicity of the antibiotic florfenicol to spe-
commercial iron oxide nanocatalysts used in ozonation process
403
19
Quantitative distribution and localization of nanoparticle-derived
Effect of 24-epibrassinolide on ros content, antioxidant enzymes and Ni
copper in whole cells of Saccharomyces cerevisiae405
accumulation in Solanum nigrum L. under Ni stress
Joint effects of nanoparticles and respective ionic counterparts
Studies on the biodegradability of two environmentally relevant
to Daphnia magna407
aliphatic organofluorines
Impact of an organic nanomaterial on soil invertebrates: sds/ddab
Phytofiltration of uranium-contaminated waters in laboratory
nanovesicles409
conditions435
431
433
Titanium dioxide nanoparticles exposure under heat shock conditions caused
a partial recovery of respiratory metabolism of Saccharomyces cerevisiae411
Distribution of Pollutants in the Biosphere
Toxicity of TiO2 nanoparticles on the gills of neotropical fish, Prochilodus
Metal(loid) allocation in the soil-plant system of a Pinus halepensis
lineatus: bioaccumulation, oxidative stress and oxidative damage
413
effects in the marine clam Ruditapes phlippinarum.415
Toxicity studies of insecticide-functionalized mesoporous silica
nanoparticles in the milkweed bug Oncopeltus fasciatus417
Effects of mesoporous silica nanoparticles on food intake and
419
Effects of ingested alumina nanoparticles (Al2O3) in the development
and reproduction of Blattella germanica421
Nanoparticles toxicity studies on microalgae R. subcapitata423
Bioremediation 425
PGMS removal from aqueous media using anaerobic bacterial communities
from WWTP sludge
425
Diversity and densities of bacteria, cyanobacteria and yeasts from waters and
sediments of estuarine areas impacted by hydrocarbon in southeast coast 427
Resistance and bioremoval of paracetamol by a sulphate-reducing
bacteria community
population growing on semiarid mine tailings
437
Levels and spatial distribution of organochlorinated compounds and
Itraq-based proteomics analysis of citrate gold nanoparticle exposure
reproduction of Blattella germanica
437
polycyclic aromatic hydrocarbons in fishes from a mediterranean coastal
lagoon439
Vertical distribution of PAHs in marine sediments (particulate fraction
and interstitial water) from eight iberian mediterranean areas
441
In-situ phytoextraction os nickel by a native population of Alyssum serpylofolium
on ultramafic sites (Portugal): prospects for agromining
443
Uranium uptake by native terrestrial and aquatic plants in an uranium
mine (central Portugal)
445
Environmental Chemistry And Toxicology
447
Repercusión de los plaguicidas utilizados en horticultura protegida
sobre el perfil tiroideo y la colinesterasa plasmática (Salto-Uruguay). 447
Estudio de los efectos del azufre y del caolín sobre Drosophila suzukii
449
Biliary PAH metabolites in european eel (Anguilla anguilla) from Mar
Menor Lagoon (SE Spain)
451
429
21
Extração assistida por micro-ondas de um bioerbicida a partir das
Gill histopathological evaluation in fish species captured in the
folhas da Canavalia ensiformis no combate da erva daninha
Douro river basin
477
Endocrine Disruption
479
Emilia sonchifolia na cultura de soja
453
Zearalenone as environmental contaminant: occurrence in broa
455
Chronic toxicity of the antiepileptic drug carbamazepine on
Free phthalate plasticizer DOPT (dioctylterephthalate) from
chemical recycling of waste pet.
457
crustacean Daphnia magna. 479
459
Monitoring and Modelling Development of novel coating free VOC´s (volatile organic
compounds) with high anti corrosion properties.
Kinetic studies and viability of using peat for removal of Al, Cu
and Pb of an effluent of a plastic recycling company
461
Risk Assessment 463
Mutagenic activity and indoor air quality in a occupational
environment of a images service of a public hospital in Brazil
463
Metais no Corpo Aquático do Riacho Mussuré e o córrego Mumbaba
e o reflexo de sua degradação na saúde dos ribeirinhos - João Pessoa
- Paraíba – Brasil
481
Modelización espacial de la contaminación por nitrato en los
acuíferos de la Cuenca Alta del río Ebro (España) mediante SIG
481
Modelización espacial de los riesgos asociados a usos del territorio
que afectan a la contaminación por nitrato en la Cuenca alta
del Ebro (España)
483
Monitorização e integração de fatores ambientais na modelação
de castas portuguesas”
485
Assessment of freshwater bivalves’ communities in Mira lagoons
487
465
Water treatment by nanofiltration and reverse osmosis: chemical
and ecotoxicological efficiency 467
Pesticide mancozeb®: derivation of risk limits for freshwater
ecosystems469
Histopathological analysis of livers from barbel (Luciobarbus bocagei),
nase (Pseudochondrostoma duriense) and trout (Salmo trutta)
from Douro basin
471
Adjusting molecular methods to the detection of toxigenic
phytoplankton genotypes in environmental and preserved samples 473
Cyanobacteria prevalence and cyanotoxin occurrence in an oligotrophic
freshwater reservoir (Beliche – Southern Portugal)
475
23
Scientific Committee
Afonso Celso Dias Bainy
Universidade Federal de Santa Catarina (Santa Catarina, Brasil)
Amadeu Soares
Universidade de Aveiro (Aveiro, Portugal)
Amparo Torreblanca Tamarit
Universitat de València (Valencia, Espanha)
Carlos Barata
Instituto de Diagnóstico Ambiental y Estudios del Agua (CSIC)(Barcelona, España)
Francisco Pereira Peixoto
Universidade de Trás-os-Montes e Alto Douro (Vila Real, Portugal)
Gisela Umbuzeiro
Universidade Estadual de Campinas (São Paulo, Brasil)
Ionan Marigomez
Universidad del País Vasco (Bilbao, España)
Julian Blasco
Inst. Ciencias Marinas Andalucia – Inst. Ciencias Marinas Andalucia (CSIC), (Cadiz, España)
Lúcia Guilhermino
Universidade do Porto (Porto, Portugal)
Luisa Castillo
Universidad Nacional (Heredia, Costa Rica)
Manuel Soto Lopez
Universidad del País Vasco (Bilbao, España)
Maria Helena Costa
Universidade Nova de Lisboa (Caparica, Portugal)
Maria João Bebiano
Universidade do Algarve (Faro, Portugal)
Nandini Sarma
National Autonomous University of Mexico (Los Reyes, State of Mexico, Mexico)
Pedro Carriquiriborde
Universidad Nacional de la Plata (La Plata, Argentina)
Vitor Vasconcelos
Ctr Interdisciplinar Invest Marinha & Ambiental (CIIMAR), Universidade do Porto (Porto, Portugal)
25
Organizing Committee
José Manuel de Melo Henriques de Almeida
Ana Catarina Gonçalves Luzio
Luis Miguel Joaquim Marques Antunes
Doctoral Researcher. University of Trás-os-Montes and Alto Douro
Ana Cristina Ramos Sampaio
Auxiliar Professor. Department of Biology and Environment. University of Trás-os-Montes and
Alto Douro
Ana Maria Monteiro Paiva Coimbra
Auxiliar Professor. Veterinary Science Department. University of Trás-os-Montes and Alto Douro
Associate Professor. Veterinary Science Department. University of Trás-os-Montes and Alto Douro
Maria Manuela do Outeiro Correia de Matos
Auxiliar Professor. Department of Genetics and Biotechnology. University of Trás-os-Montes and Alto Douro
Mário Gabriel Santiago Santos
Auxiliar Researcher. Centre for the Research and Technology of Agro-Environmental and
Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro
Auxiliar Professor. Department of Biology and Environment. University of Trás-os-Montes and Alto Douro
Anabela Ribeiro dos Reis De Castro Oliveira
Auxiliar Professor. Department of Biology and Environment. University of Trás-os-Montes and Alto Douro
Auxiliar Professor. Department of Geology. University of Trás-os-Montes and Alto Douro
Carla Maria Alves Quintelas do Amaral
Auxiliar Professor. Department of Biology and Environment. University of Trás-os-Montes and
Alto Douro
Edna Carla Janeiro Cabecinha Da Câmara Sampaio
Auxiliar Professor. Department of Biology and Environment. University of Trás-os-Montes and
Alto Douro
Fernando António Leal Pacheco
Marta Von Hafe Roboredo
Paulo Jorge de Campos Favas
Auxiliar Professor. Department of Geology. University of Trás-os-Montes and Alto Douro
Paulo José de Azevedo Pinto Rema
Auxiliar Professor. Zootechnical department. University of Trás-os-Montes and Alto Douro
Pedro Manuel de Melo Bandeira Tavares
Associate professor. Department of Chemistry. University of Trás-os-Montes and Alto Douro
Raquel Maria Garcia Dos Santos Chaves
Auxiliar Professor with Aggregation. Department of Geology. University of Trás-os-Montes and
Alto Douro
Associate Professor. Department of Genetics and Biotechnology. University ofTrás-os-Montes and Alto Douro
Francisco Manuel Pereira Peixoto
Full professor. Department of forest sciences and landscape architecture. University of Trás-osMontes and Alto Douro
Associate Professor. Department of Biology and Environment. University of Trás-os-Montes
and Alto Douro
Rui Manuel Vitor Cortes
Samantha Jane Hughes
Henrique Manuel da Fonseca Trindade
Associate Professor. Department of Agronomy. University of Trás-os-Montes and Alto Douro
Auxiliar Researcher. Centre for the Research and Technology of Agro-Environmental and
Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro
João Carlos Andrade Dos Santos
Sandra Mariza Veiga Monteiro
João Soares Carrola
Simone da Graça Pinto Varandas
Auxiliar Professor. Department of Biology and Environment. University of Trás-os-Montes and
Alto Douro
Auxiliar professor. Department of forest sciences and landscape architecture. University of
Trás-os-Montes and Alto Douro
José Júlio Gonçalves Barros Martins
Sofia Gabriel Garcia Santos
Auxiliar Professor with Aggregation. Physics department. University of Trás-os-Montes and Alto Douro
Auxiliar Professor. Zootechnical department. University of Trás-os-Montes and Alto Douro
Auxiliar Professor. Department of Biology and Environment. University of Trás-os-Montes and Alto Douro
Doctoral Researcher. University of Trás-os-Montes and Alto Douro
Solange Mendonça Leite
Auxiliar Professor with Aggregation. Physics department. University of Trás-os-Montes and Alto Douro
27
14 a 17 julho 2015
CI CTA 2015
10th Iberian and 7th Iberoamerican Congress
on Environmental Contamination and Toxicology
Scientific Topics
Aquatic Toxicology
Environmental Chemistry and Toxicology
Endocrine Disruption
Emerging Toxic Compounds (Pharmaceuticals, Agricultural contamination)
Biomarkers
Soil Pollution, Conservation and Restoration
Nanotoxicology
Aquaculture and Environment
Cell and Molecular Toxicology
Omics Analysis Methodologies
Microbial Ecotoxicology
Bioremediation
Environmental Chemistry Monitoring
Risk Assessment
Ecosystem Level Effects
Global Changes
Distribution Pollutants in Biosphere
Monitoring and modelling
Socio-economic-political aspects
Environment and Health
Programme
TUESDAY 14.07.2015
Aula Magna - 16:00 – 17:30
Opening Conference:
‘What hinders a more effective science-policy interface? How science needs to change in the age of global
environment and climate crisis’ Viriato Soromenho Marques
Environmental Pharmaceuticals in Latin America, are a relevant issue today? Pedro Carriquiriborde
WEDNESDAY 15.07.2015
Aula Magna -11.15 - 11.30
Aula Magna - 09.00 - 09.30
Keynote Session: Cyanobacteria: toxins and other secondary metabolites and their biotechnological applications | Vitor Vasconcelos
Keynote Session: Sediment toxicity tests: tools for environmental quality | Julian Blasco
09.30 - 10.45
Aula Magna
Session 1: Aquatic Toxicology I
Chairs: Vitor Vasconcelos; Rui Cortes
Histopathological baseline levels and confounding factors in
sole for marine health status assessment.
N Cuevas; Izaskun Zorita; PM Costa;
J Larreta; J Franco
Antioxidant responses and bioaccumulation in clam
Scrobicularia plana exposed to arsenate.
C Trombini; Miriam Hampel; G Rodríguez-Moro;
T García-Barrera; JL Gómez-Ariza; J Blasco
Influence of temperature on the toxicity of cadmium
and benzo(a)anthracene to planktonic microalgae.
Luis R Vieira; F Morgado; L Guilhermino
A “stock culture independent” duckweed microbiotest with
Spirodela polyrhiza: a practical and low cost alternative
to lemn24 A bioassays.
Persoone Guido; R Baudo; M Foudoulakis; G Arapis
Usefulness of fish head-kidney leucocytes to assess
the immunotoxicological effects of contaminants
Patricia Morcillo1; Héctor Cordero1; Jose Meseguer1;
María Ángeles Esteban1; Alberto Cuesta1
Auditório Geociências
Session 2: Endocrine Disruption
Chairs: Vance Trudeau; Ana Maria Coimbra
Effects of the endocrine disruptor vinclozolin in the benthic
organism Chironomus riparius (Diptera).
Mónica Aquilino; P Sánchez-Argüello;
G Morcillo; JL Martínez-Guitarte
Developmental effects of a xenoestrogen and an
aromatase-inhibitor in zebrafish (Danio rerio) embryogenesis.
Dércia Santos; M Matos; AM Coimbra
Differences in the xenoestrogenic sensitivity of five teleost
species exposed to 17α-ethinylestradiol.
A Basterretxea; G Lorenzo; C Bizarro; E Bilbao; Maren Ortiz-Zarragoitia
Gonadal development and biological recovery of zebrafish
exposed to 17ß-estradiol (E2) and 17α-ethinylestradiol (EE2).
Juliana Polloni Silva; GD de Alkimin; SC Nakahira;
CE de Souza Correa; R Fracácio1
11.00 - 12.45
Aula Magna
Session 3: Biomarkers I
Chairs: Julian Blasco; Simone Varandas
Accumulation kinetics of copper and silver and assessment of the
effects exerted after dietary exposure in oysters Crassostrea gigas.
Ane Rementeria; M Mikolaczyk; E Blanco-Rayón;
L Velasco; U Izagirre; B Zaldibar; J Schäfer
Identification of molecular biomarkers of exposure to sanitary sewage
in oyster Crassostrea brasiliana
TB Pessatti; KH Luchmann; FF Nunes; FL Zacchi;
Afonso Celso Dias Bainy; JJ Mattos
Is there endocrine disruption in male mugilids
from the Tagus estuary?
João Carrola; S Ribeiro; P Almeida; S Pedro; E Rocha; J Neto; MJ Pires; C Cruz;
F Ribeiro; C Levita; J Ferreira-Cardoso; A Fontainhas-Fernandes
Auditório Geociências
Session 4: Risk Assessment I
Chairs: Ionan Marigomez; Marta Reboredo
Maturity and stability parameters in the quality assessment of
composts, sludges and other representative organic wastes intended
to agricultural use.
Paula Alvarenga; C Mourinha; M Farto; P Palma;
J Sengo; MC Morais; AC Cunha-Queda
Rational design for safer nanomaterials: a case study with
nano-biosurfactants.
Nuno Martins; JL Pereira; CMG Duarte; LCA Dias;
FE Antunes; I Lopes
Phosphorus fractionation in sediments from a small-sized dam in a
rural mountainous catchment: case study in NE Portugal
M Von Hafe Roboredo; Anabela R Reis; JPM Pinto;
AI Oliveira; A Parker
Environment risk assessment and bioaccumulation of metals in the
metal contaminated basin.
Estefania Bonnail; AM Sarmiento; TA delValls; I Riba; JM Nieto
Evaluation of zebrafish gonad development after endocrine
disruptors’ exposures.
Ana Luzio; SM Monteiro; S Garcia-Santos; E Rocha;
A Fontaínhas-Fernandes; AM Coimbra
Lunch time
31
WEDNESDAY 15.07.2015
Aula Magna - 17:00 – 17:30
Aula Magna - 14.30 - 15.00
Keynote Session: New highlights for soil health assessment using earthworms: in vitro and molecular tools | Manu Soto
15:00 - 16:15
Aula Magna
Session 5: Ecosystem Effects and Soil Pollution
Chairs: Manu Soto; Edna Cabecinha
A new tool for water quality assessment combining
distinct levels of organization.
Rui Cortes; S Hughes; A Coimbra; J Carrola; SM Monteiro; A Pinto;
S Pereira; A Sampaio; J Jesus; V Pereira; C Santos; M Lopes; S Varandas
Inter-annual variability of soft bottom macrobenthic community
of the NW gulf of Mexico in relationship with the DWH oil spill.
D Salcedo; Luis A Soto; A Estradas; AV Botello
Free-living nematodes as bio-indicators of the DWH oil spill in the
mexican exclusive economic zone, NW gulf of Mexico.
LA Soto; Diana Salcedo; K Arvizu; AV Botello
Phytoavailability of lead in soils from an abandoned
trap-shooting range
Andrés Rodríguez-Seijo; D Arenas-Lago; M Lago-Vila;
ML Andrade; FA Vega
Auditório Geociências
Session 6: Aquatic Toxicology II
Chairs: Amparo Torreblanca; Sandra Mariza Veiga Monteigo
Interaction effects between metals and PAHs in the aquatic environment: an In vivo study on genotoxicity and DNA repair with the
zebrafish model.
Ana Silva; M Martins; MH Costa; PM Costa
Effects of personal care products (PCP) on the MXR mechanism
of zebrafish embryo.
Virgínia Cunha; K Burkhardt-Medicke; P Wellner; M Santos;
P Moradas-Ferreira; T Luckenbach; M Ferreira
Effects of multigeneration exposures of D. magna to
environmentally relevant concentrations of nickel.
Keynote Session: Use of transcriptomic tools to identify new molecular and biochemical biomarkers in aquatic
non-model organisms | Afonso Celso Dias Bainy
17.30 - 18:45
Aula Magna
Session 7: Biomarkers II
Chairs: Afonso Bainy; José Manuel Almeida
Auditório Geociências
Session 8: Risk Assessment II
Chairs: Anabela Reis; Henrique Trindade
Ecotoxicological and biochemical effects of an herbicide and a metal
on zooplankton and phytoplankton estuarine and marine species.
Wastewater reuse: a study of chloroform formation.
The influence of different microalgae diets on cell and tissue level
biomarkers in mussel digestive gland.
Carbon capture and storage (ccs) strategy: a risk assessment
overview focused on marine bacteria.
Anabela Rebelo; I Ferra; A Marques; I Gonçalves; R Oliveira; M Pereira
Filimonova Valentina; F Gonçalves; JC Marques; M De Troch; AMM Gonçalves
Esther Blanco-Rayón; M Ücker; L Garmendia; I Marigómez; U Izagirre
Sub-lethal effects of exposure to atrazine in gill cells of sea lamprey
downstream migrants.
Maria João Lança; M Machado; AF Ferreira; M Candeias; R Ferreira;
I Alves-Pereira; JS Carrola; BR Quintella; PR Almeida
Ana Rocío Borrero Santiago; I Riba López; A Del Valls Casillas
Testing potential CO2 leakages on three marine microalgae:
metal speciation and biological effects.
Esther Bautista-Chamizo; MR de Orte; IR López; A del Valls Casillas
Carlos Pinheiro; S Lopes; AMVM Soares; S Loureiro
Biomarkers, histopathology and condition indices in sole
(Solea senegalensis) exposed to contaminated sediment.
Tifanie Briaudeau; E Huerga; I Marigómez; L Garmendia;
JM Garmendia; I Zorita; U Izagirre; U Izagirre
Determination of estuarine vulnerability to contamination
for use in ecological risk assessment.
Daniel Cerqueira Ribeiro; L Guilhermino
Background concentrations of polycyclic aromatic hydrocarbons
metabolites in portuguese firemen.
M Oliveira; Klara Slezakova; A Fernandes; JA Vaz; C Delerue-Matos; MC Pereira; S Morais
33
THURSDAY 16.07.2015
Aula Magna - 9:00 – 09:30
Aula Magna - 11:00 – 11:30
Keynote Session: Pesticides in latin america: ecotoxicity | Luisa Castillo
Keynote Session: Micro - and nanoplastics as emerging contaminants in the aquatic environment: what we do – and don’t – know.
Dick Vethaak
09:30 – 10:45
Aula Magna
Session 9: Nanotoxicology
Chairs: Carlos Barata; Romeu Videira
Oxidative stress and enzymatic alterations
induced by multi-walled carbon nantoubes
(MWCNTs) funcionalized with polyethylene glycol (PEG) in mouse liver tissue.
Aud. Geociências
Session 10: Aquatic Toxicology III
Chairs: Lúcia Guilhermino; João Carrola
11:30 – 12:45
Aud. 1
Session 11: Microbial Ecotoxicology
Chairs: José Luis G Flores; Carla Amaral
Usefulness of fish head-kidney leucocytes Relative sensitivities of life history variables of
to assess the immunotoxicological effects of freshwater zooplankton (rotifers and cladocerans)
contaminants.
to stress: a review.
Patricia Morcillo; H Cordero; J Meseguer; MA Este- SSS Sarma; N Sarma
Silvia Pierre Irazusta; EC Oliveira; HJ Ceragioli; ban; A Cuesta
BF Santos De Souza; R Azevedo Jr; MA Cruz Hofling;
ZM Almeida Cruz
Effects of multi-stressors on juveniles of the In vitro inhibition by organophosphorus and
common goby (Pomatoschistus microps): mi- carbamate compounds of gilthead seabream
croplastics, nanoparticles and temperature. brain acetylcholinesterase and hepatic carboxP Ferreira; E Fonte; Lúcia Guilhermino ylesterase.
Interaction of graphene nanomaterials
with liver derived cell lines: internalization, cytotoxicity and potentiation of
chemical effects.
Effect of mixed toxic diets (Microcystis aeruginosa and Scenedesmus acutus) on the competition of two cladocerans (Daphnia cf. Mendotae
and Simocephal mixtus).
Francisco Soto-Mancera; G Albendín; J Arellano; MI Arufe
Alfredo Pérez Morales; CA Espinosa-Rodríguez;
L Rivera-De la Parra; SSS Sarma; S Nandini
Effects of carcinogenic versus non-carcinogenic
polycyclic aromatic hydrocarbons (PAHs)
and their mixtures: an integrated biomarker
survey.
Solid lipid nanoparticles affect leaf litter microbial
colonizers and enzymatic activities in freshwater.
T Lammel; P Boisseaux; Marta Martins; JM Santos; MS Diniz;
M-L Fernández-Cruz; José M Navas MH Costa; PM Costa
Ana Sampaio; RJ Mendes; PG Castro; S Doktorovova;
AM Silva
Mechanisms of action and toxicity of Influence of a seawater pulse on the struc- Effect of caffeine on the life table demography of
silver nanoparticles in model aquatic and ture of freshwater invertebrate communi- the rotifer Plationus patulus and the cladoceran
Moina macrocopa.
terrestrial organisms. ties.
Miren P Cajaraville; N Duroudier; N García-Velasco; Cátia Venâncio; C Shinn; M Abelho;
JM Lacave; A Fanjul; M Mikolaczyk; M Moreira-Santos; I Lopes; A Ribeiro; R Ribeiro
A Jimeno-Romero;
A Katsumiti; J Schäfer; E Bilbao; M Soto; A Orbea
Do cationic and hydrophobic modifications
influence the ecotoxicity of hydroxyethyl cellulose polymers?
Anabela M Simões; FE Antunes; I Lopes
Aula Magna
Aud. Geociências
Aud. 1
Session 12: Emerging Toxic Compounds Session 13: Environmental Chemistry and Session 14: Cell And Molecular Toxicology
Toxicology I
Chairs: Dick Vethaak; Luis Antunes
Chairs: Nandini Sarma ; Raquel Chaves
Chairs: Pedro Carriquiriborde; Samatha Hughes
Caracterización de la fotodegradación de Assessment of trace elements pollution and its Cytotoxicity of titanium dioxide nanoparticles
fármacos en sistemas acuáticos mediante cro- environmental risk in freshwater sediments of in fish cell lines.
matografía líquida-espectrometría de masas. the alqueva reservoir (Guadiana basin).
A Bermejo; M-L Fernández-Cruz; José M Navas
RM Baena-Nogueras; E González-Mazo; PA Lara-Martín Patrícia Palma; L Ledo; P Alvarenga
Impact of regulated and emerging pollutants The coastal lagoons of Tabasco state, Mexi- DNA damage and marine pollutants: mechaand microplastics in marine ecosystems co: endangered ecosystems.
nisms of toxicity, applications and handicaps
(impacta project). Alfonso V Botello; S Villanueva; G Ponce
of surveying genotoxicity In vivo in non-model
Víctor M León; S Muniategui; R Beiras; J Bellas; J Gago;
organisms.
JA Campillo; C Martínez-Gómez; L Viñas; V Besada;
J Fumega; JM Andrade; J Moreda; E Beceiro; R Soto; E
Alonso; MJ González; P Sánchez-Marín; L Vidal
Pedro M Costa; M Martins; MH Costa
Concerning pharmaceuticals, what are the Pentachlorophenol toxicity to a mixture Salinity effect on transcription levels in oysters
portuguese wastewater treatment plants of Microcystis aeruginosa and Chlorella Crassostrea brasiliana exposed to phenanthrene.
FL Zacchi; D de Lima; FF Nunes; JJ Mattos; KH Luchhotspots ofcontamination? vulgaris cultures.
AMPT Pereira; LJG Silva; CM Lino; Paulo de Morais; T Soichev; MCP Basto; V Ramos;
LM Meisel; A Pena VM Vasconcelos; MTSD Vasconcelos
mann; CHAM Gomes; MC Bícego; S Taniguchi;
ST Sasaki; Afonso Celso Dias Bainy
Removal of veterinary antibiotics in con- Eutrophication, water quality and nutrient Exposure to phenanthrene and depuration:
structed wetlands microcosms – response of loading: blooms of green algae.
changes on gene transcription, enzymatic
microbial community. Xana Álvarez; E Valero; J Picos
activity and lipoperoxidation in gill of scallops
Filipa Santos; CMR Almeida; I Ribeiro;
Nodipecten nodosus.
AC Ferreira; AP Mucha
RS Piazza; JJ Mattos; FF Nunes; D de Lima; FL Zacchi; R
Trevisan; AC Pessoa de Mello; MA Saldaña Serrano; IM
Martins dos Reis; CHA Miranda Gomes; M Bercht; CM
Rodrigues de Melo; Afonso Celso Dias Bainy; CE Piazza
Nandini Sarma; M C Reyes-Santillán; SSS Sarma
Aguas residuales como fuente de aporte
de piretroides en sedimentos superficiales
y organismos de Bahía Todos Santos, Baja
California, México.
Félix Augusto Hernández-Guzmán;
JV Macías-Zamora; N Ramírez-Álvarez
Developmental impairment following chronic
exposure to ketamine in early zebrafish (Danio
rerio) embryos.
Luís Félix; A Vidal; L Antunes; AM Coimbra
Lunch time
35
FRIDAY 17.07.2015
Aula Magna - 09.00 - 09.30
14 a 17 julho 2015
Keynote Session: Emerging toxic compounds.
Carlos Barata
09.30 - 10.45
Aula Magna
Auditório Geociências
Session 15: Environmental Chemistry Monitoring
Chairs: Imaculada Varó; Ana Sampaio
Microplastics in coastal sediments from south Portugal.
Jesus Gago; JPGL Frias; P Sobral; V Otero
Session 16: Environmental Chemistry And Toxicology II
Chairs: Luisa Castillo; Manuela Matos
Is metal contamination in estuarine environments endangering european eel (Anguilla anguilla) in their first stage in continental waters?
CI CTA 2015
10th Iberian and 7th Iberoamerican Congress
on Environmental Contamination and Toxicology
D Castro; Cláudia Mieiro; JP Coelho; S Guilherme;
A Marques; AC Duarte; E Pereira; M Pacheco
Polychlorinated biphenyls in molluscs, within basque estuaries
(Northern Spain): relationships with hydrological characteristics
and human pressures.
Oihana Solaun; JG Rodriguez; A Borja; J Larreta; V Valencia
Composites alto desempeño y cero voc´s para componentes automotrices y aeroespaciales, a partir de resina poliéster insaturado
dispersable en agua.
Raul Segura Cruz; MI Velazquez Sandoval; VF Gonzalez Saenz
Keynote Sessions
Multibiomarker assessment of cadmium-based quantum dots
effects in the marine mussel Mytilus galloprovincialis.
Thiago Lopes Rocha; TGomes; CCardoso; NC Mestre;
VS Sousa; MJ Bebianno
Keynote Session: Neuroendocrine disruption: more than hormones are upset by environmental contaminants |Vance L Trudeau
14.30 - 15.30
Aula Magna
Session 17: Bioremediation
Chairs: Maria João Bebianno; Solange Leite, João Santos
Synergistic and beneficial effects of 2,4-epibrassinolide and
progesterone for the environmental remediation of
progesterone by Solanum Nigrum L. Plants.
Ana Pinto; C Soares; F Fidalgo; J Teixeira
Effect of aluminum on ROS content and antioxidant system
in rye (Secale cereale L.) leaves and roots.
Session 17: Bioremediation
Chairs: Maria João Bebianno; Solange Leite, João Santos
Bioremediation of different types of oil in estuarine and coastal
environments – the role of autochthonous microorganisms.
Vanessa Gouveia; CMR Almeida; AP Mucha
An ecological relevant approach to oil sands.
Diogo N Cardoso; FJ Wrona; AMVM Soares; S Loureiro
Alexandra de Sousa; M Matos; H AbdElggawad;
H Asard; J Teixeira; F Fidalgo
Aula Magna 15:15 – 16:30
Closing Conference:
Effect models at the core of ecotoxicology: insights and implications. Roman Ashauer
How human wellbeing is affecting the aquatic environment. Maria João Bebianno
Opening conference (14.07 – 16:30)
Environmental sustainability. Insights to the future.
‘What hinders a more effective science-policy interface? How science
needs to change in the age of global environment and climate crisis’
Viriato Soromenho Marques
There is no easy transition from science to policy making. Complexity of issues,
plurality of actors, antagonistic agendas and the uncertainties in the social role and
status of science are among the most visible -- although with deep hidden features obstacles to an effective science-policy interface. The case of climate change works like
a kind of magnifying lens allowing us to exercise a more acute critical judgement upon
the constellation of problems we may identify in this crucial area of public policy framing.
Viriato Soromenho-Marques (1957) teaches Political Philosophy and European Ideas in the
Departments of Philosophy and European Studies of the University of Lisbon, where he is Full
Professor. Since 1978 he has been engaged in the civic environmental movement in Portugal
and Europe. He is member of the National Council on Environment and Sustainable Development
(since 1998). He was Vice-Chair of the European Environmental and Sustainable Development
Advisory Councils network (2001-2006). He was one of the authors of the Portuguese National
Strategy for Sustainable Development (2004). Former member of the Advisory Group on
Energy and Climate Change from the EC President (2007-2010). Scientific Coordinator of the
Environment Program from Calouste Gulbenkian Foundation (2007-2011). Member of the
Lisbon Academy of Sciences and also of the Marine Academy. More information regarding his
bibliography and activities may be find on: www.viriatosoromenho-marques.com
39
Environmental Pharmaceuticals in Latin America, are a relevant
issue today?
Pedro Carriquiriborde1
The scientific concern about the potential environmental impacts of
pharmaceuticals and personal care products on aquatic ecosystems has risen since
the beginning of the new century. However, in Latin America this issue still is lagged.
Most of the cities still dump raw, or luckily primary treated, wastewaters to rivers
and lakes. However, the occurrence profile and concentrations of pharmaceuticals
in wastewater effluents and receiving waters are not far from those reported for
other regions of the world. Studies with local fish species, for example, showed
lethal acute toxicity levels for NSAIDs, psychiatric/anticonvulsants and antibiotics,
frequently observed in wastewaters, comprised within moderately to slightly toxic.
In the case of sublethal responses, the effects depend on the pharmaceuticals and
the assessed endpoints. Generic biomarkers seems no to be informative, but some
specific responses are sensitive enough to detect effects at concentrations similar
to that observed in receiving waters. However, levels of organic matter, dissolved
oxygen and ammonia found in many places of Latin American close to discharges
still hide the environmental relevance of pharmaceuticals.
1Centro
de Investigaciones del Medio Ambiente (CIMA), Facultad de Ciencias Exactas, Universidad
Nacional de La Plata, Calle 47 y 115 s/n (1900) La Plata, Buenos Aires, Argentina. Tel/Fax +54 221 4229329
41
WEDNESDAY 15.07.2015 – 09:00
Keynote Session: Vitor Vasconcelos
Cyanobacteria: toxins and other secondary metabolites and their
biotechnological applications.
Vitor Vasconcelos 1,2
Cyanobacteria are amongst the oldest organisms living on our planet and were
responsible for the generation of an oxygen rich atmosphere and the first to harvest
energy from light via photosynthesis. Because of this they are on the origin of plants,
one of the most commonly used groups of organisms from which novel biomolecules
have been extracted and used as pharmaceutical products and/or compounds. Due
to the old origin of cyanobacteria, they have unique genomes, many of which are now
unravelled, that are responsible for the production of a high diversity of secondary
metabolites with a high range of biological activities. In the second half of last
century, cyanobacteria secondary metabolites were mostly studied due to their toxic
properties, in special to mammals, resulting in the discovery of metabolites such as
the peptides microcystins and nodularins or the alkaloids anatoxin-a, saxitoxins (also
produced by dinoflagelates) and cylindropermopsin (Moreira et al., 2014). More
recently marine cyanobacteria were also found to produce the neurotoxin palytoxin,
formerly known as being produced by dinoflagelates Most of the cyanobacteria
toxins are produced non-ribossomally by clusters of genes comprising polyketide
synthases (PKS) and nonribosomal peptide synthetases (NPRS) but in recent years,
cyanobacteria have also been found to produce other bioactive compounds, such
as cyanobactins, anabaenolysins, hassallidins portoamides and hierridin-B (Leão
et al., 2013). These compounds can help to develop more environmental friendly
43
products for involved industries, thanks to new processes to isolate, characterize and
WEDNESDAY 15.07.2015 | 11.00 - 11.30
mass-produce them. Cyanobacteria natural products have a variety of reported
Keynote Session: Julian Blasco
activities, being the anticancer the most studied (Costa et al., 2014). Nevertheless,
other less reported activities such as antiviral, antifungal, anti-parasitical as well as
application (i.e. anti-fouling, allelopathic and cosmetics) although less studied have
an enormous potential (Almeida and Vasconcelos, 2015).
Sediment toxicity tests: tools for environmental quality.
Julián Blasco1, Olivia Campana2, C.V. Araujo3, Araceli Rodríguez-Romero1, I. Moreno-Garrido1
Sediments represent the main sink for pollutants and depending on environmental
conditions they can be a source of contaminants to water column. On the other hand,
CIIMAR/CIMAR - Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua
dos Bragas 289, P 4050-123 Porto, Portugal; 2Faculty of Sciences, Porto University, Rua do Campo Alegre,
4169-007 Porto, Portugal;
1
sediments are a component of aquatic ecosystems providing feeding, rearing and spawning
areas for multitude of organisms. To establish the quality of sediments and the behavior
of contaminants is a challenge for assessing environmental quality and the health of
ecosystems. Sediment toxicity testing and bioassays have been defined in restrictive way and
they can be applied using different approaches (e.g. evaluate the effect of the contaminants
after spiking to artificial or natural sediments or evaluate the effect of the environmental
contamination collecting sediments from the field). In both cases, the objective is getting
a deeper insight into the biological effects of spiked contaminants or pollutant mixture,
respectively. In this presentation both approaches will be considered under the umbrella
of sediment toxicity testing. The use of uncommon species (phytobenthos), behavioral
response, the effect of geochemical properties to biological responses, the development
of sediment quality guidelines and the employment of sediment toxicity tests for assessing
the acidification process on metal mobilization have been examined. In summary, sediment
toxicity testing is an essential tool to establish the environmental quality of ecosystems and
to define strategies for their management in a sustainable way.
1Instituto de Ciencias Marinas de Andalucía (CSIC). Campus Río San Pedro. 11510 Puerto Real (Cädiz). Spain
2Environment Department. University of York. York, YO10 5DD, UK
3Centre for Functional Ecology (CFE), Department of Life Sciences, University of Coimbra, Calçada Martim
de Freitas, 3000-456 Coimbra, Portugal
45
WEDNESDAY 15.07.2015 | 14.30 - 15.00
Keynote Session: Manu Soto
New highlights for soil health assessment using earthworms: in vitro
and molecular tools.
Manu Soto1,2
Alteration of Biosphere by anthropogenic activities is significant and constantly
rising enhancing the contaminant loads in the ecosystems in general, and in soils
in particular. Therefore, the global concern on soil pollution has grown during the
last decades, and consequently the assessment of its effects on soil ecosystems has
become a priority issue. The potential risk of pollutants has been mainly studied in
aquatic environments, while their effects in soils are poorly investigated despite the
great complexity of soil matrix and the potential interactions of soil components
with pollutants that could affect their toxicity. Different evaluation strategies have
been proposed to assess soil health; among them the use of terrestrial sentinels is
undeniable. Earthworms, key species in soil ecosystems, are one of the most studied
sentinel taxa since their pollutant body burdens reflect environmentally bioavailable
pollutant levels and measurable responses (biomarkers) can be quantified in them.
Among earthworms, Eisenia fetida is a model species in terrestrial ecotoxicology
that has been broadly used in standardized OECD toxicity tests to assess adverse
effects regarding survival, reproduction rate and/or growth (OECD 207, OECD 222).
However, these approaches are limited when the amount of soils to be tested is large
and when a rapid assessment is required. Therefore, there is a need to develop fast
and accurate methodologies for rapid screening diagnosis of contaminated soils,
understandable for decision makers. In this framework, biomarkers measured at
47
different levels of biological complexity give indication of soil health in an integrative
WEDNESDAY 15.07.2015 | 17:00 – 17:30
and comprehensive manner. Moreover, the potential of in vitro techniques and “omics”
Keynote Session: Afonso Celso Dias Bainy
technologies is beginning to provide promising applications in this field. Hence,
a strong effort is being carried out (a) to perform extensive and high throughput
toxicity testing based on the assessment of biological effects, and (b) to develop an
efficient and cost-effective soil health assessment strategy that will provide straight
forward advice and support to stakeholders involved in environmental protection.
Use of transcriptomic tools to identify new molecular and biochemical
biomarkers in aquatic non-model organisms
Afonso C.D Bainy
The aquatic contamination arises from industrial, agricultural, mining,
1CBET Research Group, Dept. Zoology and Animal Cell Biology;
2Research Centre for Experimental Marine Biology and Biotechnology PIE-UPV/EHU, University of the
Basque Country UPV/EHU, Basque Country
sanitary sewage effluents, among others, which are continually discharged in the
aquatic environment, frequently without proper wastewater treatment. Effects
of contaminants to aquatic organisms can be assessed by using early warning
molecular biomarkers, which usually have low ecological relevance, but can help
to prevent more serious damage to the ecosystem. Different approaches have
been proposed for the identification of molecular biomarkers. Next-generation
sequencing (NGS) techniques have been used to identify hundreds to thousands of
differentially transcribed genes, through RNA sequencing. Understanding the global
transcriptional pattern of exposed organisms can help environmental agencies
to establish more biologically-relevant programs for assessment or monitoring.
Analyzing genes that are repressed or activated under different situations of
environmental contamination can help to identify more resistant and sensitive
populations. Since 2003, we have been working in identifying differentially transcribed
genes in Pacific oysters (Crassostrea gigas), mangrove oysters (Crassostrea brasiliana
and Crassostrea rhizophorae) and fish Brazilian guppy (Poecilia vivipara) exposed to
different complex mixtures, such as untreated sanitary sewage, aimed to identify new
biomarkers of contamination into the marine environment. Special attention has
been given to the identification and validation of genes related to Phase I and Phase
49
II Biotransformation of xenobiotics, such as Cytochrome P450 isoforms, Glutathione
THURSDAY 16.07.2015 | 11:00 – 11:30
S-transferases and Sulfotransferases. It is important to consider that changes in gene
Keynote Session: Dick Vethaak1
expression are not necessarily proportional to the actual biological response, in terms
of protein synthesis and/or enzyme activity. Whenever possible it is recommended to
incorporate functional studies by analyzing two-dimensional protein expression, or
by evaluating the activity of key enzymes of the biotransformation systems, in order
to verify whether certain pathways might or might not be compromised. In this way
we can certify whether the observed responses show adaptability or susceptibility to
contaminants. Supported by CNPq, INCT-TA, Petrobras S.A.
1Laboratory of Biomarkers of Aquatic Contamination and Immunochemistry (LABCAI) Department of
Biochemistry, Federal University of Santa Catarina (UFSC) Florianópolis, SC 88040900, Brazil - e-mail:
afonso.bainy@ufsc.br
Micro and nanoplastics as emerging contaminants in the aquatic environment: what we do and don’t know.
While scientific evidence illustrates the presence and potential dangers of
microplastics (synthetic polymers with a diameter smaller than 5 mm to the nanoscale
range) in the marine environment, much must still be elucidated on the mechanisms
and modes of toxic action and which kinds of species are at the greatest risk of being
adversely affected. What makes microplastics very different compared to other
aquatic pollutants is their degradation half-lives are in the order of hundreds of years
– many times longer than even the persistent organic pollutants (POPs). In addition,
plastic particles might be a vector for hazardous substances because they can sorb
persistent, bioaccumulating and or toxic chemicals (e.g. POPs, endocrine disrupters).
Thus, plastic particles may facilitate the entrance of these substances into organisms
and the food chain, potentially threatening human health both chemically and by
particle toxicity.
A wide variety of marine and freshwater organisms have now been found to
ingest microplastics, including invertebrates, fish, birds and mammals. Laboratory
studies demonstrate that ingested microplastics can affect the physiology of the host
organism and potentially compromise its fitness. The translocation, bioaccumulation,
and elimination of plastic particles by aquatic organisms will depend on the size of
the particle. Particles at the smaller end of the size spectrum (nanoscales) have been
shown to cross membranes under controlled laboratory conditions, causing tissue
damage.
51
This presentation gives a brief overview of the state of knowledge regarding
the exposure pathways, occurrence, fate and effects of nano- and microplastic
FRIDAY 17.07.2015 | 9:00 – 09:30
Keynote Session: Carlos Barata
pollution in the aquatic environment. Special attention is given to the following
topics: key sources and types of micro- and nanoplastics; impacts and effects
observed in laboratory and field studies; risk for aquatic organisms and human
Emerging toxic compounds.
consumers of marine foodstuffs. Examples from the literature will be extended
with our own research, including the occurrence of plastic particles in wastewater
effluents, environmental samples and invertebrates from the North Sea, Dutch rivers
and Amsterdam canal zone; and laboratory experiments with microalgae and sea
urchins. Lastly, major knowledge gaps are identified and challenges to improve a
future assessment are proposed.
1Institut for Environmental Studies (IVM), VU University Amsterdam
Whereas the chemical identification of pollutants has advanced notoriously in
recent years, allowing quantification of a wide array of potential pollutants in the
environment at sub-ppb levels, the biological characterization of their putative effects
on ecosystems and human health is clearly lagging behind both in terms of defining
new risks (“emerging pollutants”), identifying emerging effects of classical pollutants
(behaviour, neurological, immunological or metabolic disorders, for example) or
characterizing the mechanism of toxicity. This is a most serious impediment for both
the correct evaluation of environmental impacts and for the assessment of risks
associated to new substances. In this talk I will present recent findings from our lab
to uncover new potential modes of action of classic and emergent contaminants
and manufactured nanoparticles across species. I will focus in biological processes
for which there is evidence of pollution impact but that have not been yet studied
in depth. Such biological processes include non-conventional endocrine pathways
(those modulated by thyroid hormones, retinoic acid, ecdysteroids, terpenoids),
developmental neurotoxicity, neuroendocrine disorders in invertebrates, oxidative
stress and effects on the multixenobiotic resistant mechanism. Adverse effects
of
different micropollutants (bisphenol-A, organohalogenates, pesticides,
manufactured nanoparticles (NPs),pharmaceuticals and personal care products)
will be reported across different levels of organization (transcriptional, biochemical,
phenotypic and populational) using several toxicological models (Danio rerio,
Daphnia magna, zebra mussel. Nobel technological and conceptual developments
53
such as the use and application of non target analyses into analytical chemistry
FRIDAY 17.07.2015 | 11:00 – 11:30
and omics and the application of adverse outcome pathways to study emerging
effects and assess mechanism of action of chemicals in the lab and in the field will
Keynote Session: Vance L., Trudeau
be discussed.
Neuroendocrine Disruption: more than hormones are upset by
environmental contaminants
Acknowledgement
This work has been funded by the Spanish projects CGL2008-01898, CTM2011-30471-C02-01 and
CTM2014-51985-R.
1Institut of Environmental Assessement and Water Research (IDAEA), Spanish Council of Research (CSIC), Jordi
Girona 18, 08034 Barcelona
Industrial pollutants, agricultural pesticides and pharmaceuticals are now
ubiquitously present in the environment. Some but not all may pose a risk to aquatic
vertebrates. Contamination can be detected at all levels, from water to soil to air
to animal and human tissues. We have proposed that the term ‘neuroendocrine
disruption’ extends the concept of endocrine disruption to include the full breadth
of integrative animal physiology. Disruption to homeostatic mechanisms by
environmental pollutants can affect an animal’s ability to undergo reproduction or
develop normally, and may lead to transgenerational deficits. Complex effluents from
pulp and paper processing contain an array of neuroactive phytochemicals that may
lead to disrupted spawning, largely as a result of female-specific neuroendocrine
effects. Antidepressants such as fluoxetine are environmental contaminants that
disrupt reproduction, behaviour and metabolism in teleost fish, demonstrating the
complex and pervasive effects of neuroactive pollutants. Therefore, waterborne
pollutants have a range of effects on neuroendocrine systems that may not involve
classic toxicological responses.
Much like endocrine disruption is very different
from classical toxicology, neuroendocrine disruption is distinguishable from
neurotoxicology. Neurotoxicologists may study chemical insults and mechanisms
underlying subsequent cell death in the central or peripheral nervous system, which
can eventually lead to the failure of key regulatory systems, compromised health
and perhaps the death of exposed individuals. In contrast, the consequences of
55
disrupting the complex neurohormonal brain-pituitary-target organ communication
FRIDAY 17.07.2015 | 15:15 – 15:45
systems are within the domain of neuroendocrine disruption.
Keynote Session: Roman Ashauer
Supported by NSERC and Ottawa Research Chair program.
Effect models at the core of ecotoxicology: insights and implications.
Dept. Biology, University of Ottawa, ON, Canada, K1N 6N
At the core of ecotoxicology must be a conceptual idea of how chemicals affect
organisms. The talk will explore how explicitly accounting for toxicokinetic and
toxicodynamic processes enables a better understanding of organism recovery,
modes of action and the interplay of hidden assumptions and conclusions about
toxicity.
I will demonstrate that simple organism level endpoints, such as mortality,
already contain information about the mode of action and organism recovery. After
an overview of how to extract that information I will show how this opens new
possibilities for chemical safety testing and assessment. Essentially toxicodynamic
parameters exhibit patterns in chemical space. This important insight can be
leveraged to identify the mode of action of new chemicals, to predict toxicity of
untested chemicals with known modes of action and to facilitate in vitro to in vivo
toxicity extrapolation.
The last third of the talk will discuss the challenge of biotransformation
across species and chemicals and the relative importance of toxicokinetics and
toxicodynamics. Finally an application of toxicokinetics for reverse dosing will be
shown which enables the accurate predication of a sub-lethal endpoint in fish from
in vitro data. The implications for ecotoxicology are that toxicokinetics are in many
cases more important to solve today’s challenges than toxicodynamics and that
simple toxicodynamic models yield powerful results.
57
FRIDAY 17.07.2015 | 15:45 – 16:15
How human wellbeing is afecting the aquatic environment
Maria João Bebianno1
Oceans are connected to human health and wellbeing but the impact of
humans on ocean health is changing this paradigm. The continuous introduction
in the last two decades of compounds used to increase human wellbeing such as
pharmaceutical and personal care products (PPCPs) in the oceans is posing a serious
risk on the ocean health particularly in coastal areas. However, the knowledge of
the mechanisms and effects of the presence of these compounds in the ocean is
poorly understood. In the case of pharmaceutical compounds, UV filters and of their
mixture it is already recognised that these compounds are accumulated in aquatic
organisms, distributed among tissues and enter the cell causing in some cases
similar action of those in humans. Therefore, it becomes essential to define the best
strategies to detect and characterize the presence of these stressors in the aquatic
environment and select the most appropriate biological effects to clearly assess their
ecological risk along with measures to diminish their inputs. Examples will be given
on the impact and presence of pharmaceutical compounds and UV filters in marine
molluscs either exposed in the laboratory or present in coastal areas. Therefore, it is
crucial to assess how marine organisms exposure of these emerging contaminants
present in the oceans may return to humans and affect human health.
1CIMA, University of Algarve, Campus de Gambelas, 8000-139 Faro, Portugal
mbebian@ualg.pt
Environment Department, University of York, UK
59
14 a 17 julho 2015
CI CTA 2015
10th Iberian and 7th Iberoamerican Congress
on Environmental Contamination and Toxicology
Oral Communication
61
Aquatic Toxicology
Histopathological baseline levels and confounding factors in sole for
marine health status assessment
Nagore Cuevas1; Izaskun Zorita1; Pedro M. Costa2; Joana Larreta1; Javier Franco1
Fish histopathology has become an important tool in aquatic toxicology, having
been implemented in many biomonitoring programmes worldwide. However,
there are various gaps in the knowledge of many sentinel organisms and the
interference of confounding factors. This work aimed (i) to determine baseline levels
of hepatic and gonadal histopathological traits and hepatic metal bioaccumulation
levels in soles (Solea solea) collected monthly during one year along the Basque
continental shelf (SE Bay of Biscay), a presumably pristine area and (ii) to establish
how confounding factors may influence the histopathological assessment. Results
indicate that biometric parameters varied according to age, reproductive cycle and
gender. The maximum peak in length, weight and gonadosomatic index was shown
in females during the spawning period while a decreasing trend was observed
afterwards. This decrease was consistent with the highest Cd, Cu and Hg levels
recorded in the liver at the post-spawning period in both genders. Additionally, a Zn
peak was shown in pre-spawning females probably associated with vitellogenesis.
Fat vacuolation and spongiosis hepatis were the most prevalent hepatic lesions.
As expected, fat vacuolation showed the highest frequencies at the pre-spawning
period, while necrosis and melanomacrophage frequencies increased at the postspawning period probably as a result of reabsorption activities. Nevertheless, when
all the alterations were integrated in the liver histopathological index no significant
differences were detected among seasons or between genders. Liver endured more
63
severe alterations than gonad. However, gonad histopathological index in females
Aquatic Toxicology
was more variable during the year and presented higher values than in males,
maybe due to the morphometric changes associated with the reproductive cycle. In
In vitro inhibition by organophosphorus and carbamate compounds of
conclusion, these findings provide the knowledge to differentiate pollution-induced
gilthead seabream brain acetylcholinesterase and hepatic carboxyles-
effects from the natural physiological responses in soles from the Basque continental
terase
shelf, supporting the use of fish histopathology in marine health status assessment.
F. Soto-Mancera1; G. Albendín1; J. Arellano1; M.I. Arufe1
1AZTI;
2MARE-Universidade Nova de Lisboa
Organophosphorus (OPs) and carbamate (CBs) pesticides cause inhibition
of acetylcholinesterase (AChE) and carboxylesterase (CbE) by, respectively,
phosphorylating or carbamilating a serine hydroxy group at their active site. In the
case of AChE, this causes accumulation of the enzyme at synaptic cleft and results in
overstimulation of cholinergic pathways and subsequently neurotoxic effects. CbE
and, above all, AChE inhibition has been widely used as biomarker of OP and CB
pesticide exposure. However, it has been shown, in some studies, that CbE could
be more sensitive to these inhibitors than AChE. Due to that fact, the combined use
of these two enzymes together has been suggested as a good monitoring strategy.
Furthermore, OPs and CBs seem to bind stoichiometrically to carboxylesterases,
so it has been reported that these enzymes could have a protective role against
AChE inhibition by these pesticides. The in vitro studies include the determination
of pesticide concentration (or active metabolite) that inhibits the enzyme by 50%,
i.e., IC50. To provide basal information for environmental monitoring in coastal and
marine areas, biochemical studies were conducted to compare the in vitro sensitivity
of the gilthead seabream brain AChE and hepatic CbE to inhibition by chlorpyrifosoxon, the active metabolite of the phosphorothioate insecticide chlorpyrifos, the
active OP dichlorvos, and the carbamate physostigmine. Because of the possible
existence of different CbE isozymes, several substrates (4-nitrophenyl acetate,
65
4-nitrophenyl valerate, and 1-naphthyl acetate) were used to measure the CbE
Aquatic Toxicology
activity. Physostigmine inhibited brain AChE, but it hardly inhibited hepatic CbE. On
the other hand, chlorpyrifos-oxon caused a greater AChE and CbE inhibition than
Usefulness of fish head-kidney leucocytes to assess the immunotoxico-
dichlorvos. Finally, the CbE inhibition caused by dichlorvos was higher than the AChE
logical effects of contaminants
inhibition, whereas Chlorpyrifos-oxon caused similar IC50 values in both enzymes,
Patricia Morcillo1; Héctor Cordero1; Jose Meseguer1; María Ángeles Esteban1; Alberto Cuesta1
brain AChE and hepatic CbE. No differences were found in IC50 values by using
different CbE substrates.
Exposure to certain heavy metals has been shown to disturb, among others,
immune responses in a variety of fish species, produce mortality and alterations
1Toxicology
Laboratory. Faculty of Marine and Environmental Sciences. University of Cádiz, CASEM,
Polígono Río San Pedro, 11510, Puerto Real, Cádiz (Spain).
in nutrition and reproduction. However, few studies have evaluated the
immunotoxicological role of aquatic contaminants in vitro, and they could represent
good models for immunotoxicological studies. Thus, we have in vitro exposed
gilthead seabream (Sparus aurata) and European sea bass (Dicentrarchus labrax),
the most important farmed fish species in the Mediterranean area, head-kidney
leucocytes (HKLs) to cadmium (Cd), methylmercury (MeHg) and arsenic (As) and
evaluated the immunotoxicological impact. HKLs cell death (apoptosis and necrosis),
oxidative stress, phagocytosis, respiratory burst activities and gene expression were
determined. Our data showed that short exposure (from 30 min to 2 h) to Cd, MeHg
or As promoted apoptosis and oxidative stress. In addition, heavy metals differently
altered the HKLs innate immune activities as well as the immune-related gene
expression. Further studies are needed to ascertain how heavy metals affect the
immune system of marine fish and the mechanisms involved.
University of Murcia
1
67
Aquatic Toxicology
Effects of carcinogenic versus non-carcinogenic polycyclic aromatic hydrocarbons (PAHs) and their mixtures: an integrated biomarker survey
Marta Martins1; José M. Santos1; Mário S. Diniz2; Maria Helena Costa1; Pedro M. Costa1
Polycyclic aromatic hydrocarbons (PAHs) are organic pollutants that invariably
occur in the aquatic environment as mixtures tending to be trapped in aquatic
sediments due to their high hydrophobicity. Due to their acknowledged toxicity and
carcinogenic potential, PAHs are regarded as priority substances, being included
in the EU’s Priority Substances Directive, being also classified as non-carcinogenic,
potentially carcinogenic and effectively carcinogenic to humans by the International
Agency for Research on Cancer (IARC). Nonetheless, the differential toxicological
effects and mechanisms between the various classes of PAHs and their mixtures
are scarcely known, especially under ecologically-relevant scenarios. The present
work aimed at understanding the toxicological mechanisms of carcinogenic and
non-carcinogenic PAHs in a model marine fish (the seabass Dicentrarchus labrax),
through the integration of genotoxicity, biochemical and histopathological
biomarker analyses. For the purpose, 28-day laboratory assays were conducted with
sediments spiked with ecologically-relevant concentrations of two priority PAHs,
isolated and combined: the “carcinogenic” benzo[b]fluoranthene (B[b]F) and the
“non-carcinogenic” phenanthrene (Phe). The results showed that, whereas exposure
to isolated Phe yielded biochemical changes better related to oxidative stress, B[b]
F disrupted metabolic responses and caused higher clastogenic/aneugenic effects,
which, altogether represent the most severe DNA damage, comparatively to strand
breakage (determined through the comet assay) . On the other hand, mixtures
69
yielded effects and responses compatible with the AHR pathway, generating
Aquatic Toxicology
additive, if not synergistic, effects, in accordance with elevated DNA lesions. Both
PAHs induced hepatic histopathological changes in liver (the main detoxification
Interaction effects between metals and PAHs in the aquatic environ-
organ) indicating metabolic failure and inflammation, especially in animals exposed
ment: an in vivo study on genotoxicity and DNA repair with the zebrafish
to mixtures. Nonetheless, the low, ecologically-relevant, concentrations of PAHs
model
diluted dose and time-effect relationships. Overall, although seemingly predicting
Ana Silva1; Marta Martins1; Maria H. Costa1; Pedro M. Costa1
the risk of individual PAHs, environmental guidelines may not apply to mixtures by
underestimating adverse effects, which calls for a redefinition of standards when
determining the true risk of toxicants under realistic scenarios.
Higher molecular weight polycyclic aromatic hydrocarbons (PAHs) like benzo[a]
pyrene (B[a]P) are potent genotoxicants through the generation of adduct-forming
metabolites via CYP1A bioactivation. Also, bioactivation generates reactive oxygen
1MARE - Marine and Environmental Sciences Centre;
2UCIBIO - Departamento de Química, FCT-UNL
species (ROS), able to oxidise nucleobases. Conversely, Cd is a weak mutagen on
its own but presumably a potent co-mutagen able to hinder DNA repair enzymes
involved in either base excision repair (BER) for the removal of oxidised bases or
nucleotide excision repair (NER) to remove adducts. In order to understand the
interaction effects of these ubiquitous aquatic toxicants onto genotoxicity and
DNA repair, laboratory-brood zebrafish [GFP-tagged strain Tg(beta-actin:HRASEGFP)] were exposed under strict laboratorial conditions to environmentallyrelevant concentrations of either toxicant and their combination (100 mg/L Cd and
500 ng/L B[a]P) for 14 days, followed by a depuration period of 7 days to survey
the recovery from DNA damage after insult. The animals were sampled at days 7,
14 and 21. The Comet assay, standard and enzyme-modified with the enzyme
formamidopyrimidine DNA glycosylase (FPG) to detect oxidative damage through
the removal of 8-oxoguanine, one of the most common forms of oxidised bases,
was employed to detect DNA strand breakage. The erythrocytic abnormalities assay
(ENA) was also performed to assess clastogenic/aneugenic damage. Either assay
was performed in whole-blood and integrated with more traditional endpoints to
71
detect, for instance, lipid peroxidation and histopathological lesions, in the gills, for
Aquatic Toxicology
being the main organ of apical entry. The results show that the FPG-modified Comet
assay is effective in vivo and able to provide a distinction between oxidative and-
Effects of personal care products (PCP) on the MXR mechanism of
non oxidative DNA damage induced by the toxicants. Also, the findings yielded a
zebrafish embryos
complex pattern of alterations and responses to toxicological challenge that show
V. Cunha1,2; K. Burkhardt-Medicke3,6; P. Wellner3; M. Santos1,4; P. Moradas-Ferreira2,5; T. Luckenbach3; M. Ferreira1
that metal:PAH interactions in vivo may compromise the notion that exposure to
environmentally-relevant concentrations of the toxicants yield only null or transient
results.
The Multixenobiotic resistance (MXR) mechanism, mediated by ABC (ATP binding
cassette) transport proteins, is present in aquatic species and acts as a cellular
defence protecting the organisms against natural and man-made toxic compounds
1 MARE - Marine and Environmental Sciences Centre, Departamento de Ciências e Engenharia do Ambiente,
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa, 2829-516 Caparica, Portugal
in the water. However, some chemicals, so called chemosensitizers, inhibit these
transport proteins increasing the sensitivity of cells and organisms to toxic chemicals
that normally are effluxed from the cells. In zebrafish, Abcb4 has similar functional
properties as ABCB1 in mammals and is considered to be the first line of defence
against toxicants. The aim of this work is to determine chemosensitizer effects of
chemicals on fish ABC transporter activity, using zebrafish embryos and assess
the interaction of different Personal Care Products (PCPs)with Abcb4 activity.
ABC transporter activity was determined by means of accumulation assays using
a fluorescent substrate, rhodamine 123 (RH123), in 24 hours post fertilization
embryos exposed to different PCPs, such as propylparabene, isoeugenol or musk
xylene, among others. For assaying the transporter ATPase activity cell membrane
preparations with highly enriched transporter protein (Abcb4) were used. The
transporter ATPase activity is indirectly determined by measuring the amount of
inorganic phosphate released during the hydrolysis of ATP. Interaction of compounds
with Abcb4 was assayed in two ways in the ATPase assay: stimulation of basal activity
and inhibition of stimulated ATPase activity. Preliminary results from accumulation
assays with fish embryos revealed interactions between the tested compounds and
73
the MXR system. Abcb4 ATPase activity results showed an inhibition of activity by
Aquatic Toxicology
PCPs, such as isoeugenol, and stimulation by nerol and musk xylene. In conclusion
some PCPs can act as chemosensitizers of efflux proteins of zebrafish showing the
Influence of a seawater pulse on the structure of freshwater invertebrate
importance of monitoring the presence of these substances in the environment.
communities
Cátia Venâncio1; Cândida Shinn2; Manuela Abelho3; Matilde Moreira-Santos4; Isabel Lopes1; Ana
Ribeiro4; Rui Ribeiro4
Acknowledgments
V. Cunha was supported by a FCT fellowship (SFRH/BD/77931/2011) and the study was funded by PEst-C/
MAR/LA0015/2013.
1CIIMAR/CIMAR—Interdisciplinary Centre of Marine and Environmental Research, Laboratory of
Environmental Toxicology, University of Porto, Rua dos Bragas, 289, 4050, Porto, Portugal;
2ICBAS/UP—Institute of Biomedical Sciences Abel Salazar, University of Porto, Largo Professor Abel
Salazar, 2, 4099, Porto, Portugal;
3UFZ—Helmholtz-Centre for Environmental Research, Department of Bioanalytical Ecotoxicology,
Permoserstraße 15, 04318 Leipzig, Germany;
4FCUP—Dept of Biology, Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169, Porto, Portugal;
5IBMC—Institute for Molecular and Cell Biology, University of Porto, Rua do Campo Alegre, 823, 4150-180
Porto, Portugal;
6Technische Universitaet Dresden, Faculty of Environmental Sciences, Institute of Hydrobiology, 01062
Dresden, Germany
Many coastal freshwater ecosystems are already facing seawater intrusions and
recent projections predict a worst scenario of 80 cm rise by the end of 2100.
Salinity is an important abiotic factor influencing communities’ abundance,
structure and composition by, for example, influencing relationships among
populations that compose a community. Accordingly, the present work intended
to evaluate the long-term effects of a pulse of seawater intrusion in the structure
and composition of a freshwater invertebrate community simulated in outdoor
mesocosms.
Model outdoor ecosystems (mesocosms) were pulsed-contaminated (0, 2.02, 3.34,
5.51, 9.09, and 15.0 mS cm-1) with different loads of natural seawater, collected in the
Atlantic Ocean (40° 9´10´´N 8°52´22´´W) (52.7 mScm-1). Three distinct periods were
established: saltwater intrusion (SI, 33 days), start of recovery with periodic dilutions
(RD, 58 days) and recovery from end of dilution onward (RPD, 126 days). Physical,
chemical and biological parameters were monitored.
The most common macroinvertebrate taxonomic groups identified were Diptera,
Ephemeroptera and Mollusca. Diptera showed the highest abundances in the control
and at salinity 15, in the RPD phase and Mollusca appeared at salinities 9.1 and 15
mScm-1, only during the RPD phase. Ephemeroptera did not recover from seawater
intrusion at concentrations of 15 mScm-1.
Zooplankton most abundant groups were, in decreasing order, Rotifera,
75
Copepoda, and Branchipoda. during SI phase. The most representative groups
Aquatic Toxicology
during SI were Rotifera and Copepoda. Despite the decrease in organism abundance
during SI, a slight increased in organism abundance was observed at the end of this
Effects of multigeneration exposures of D. magna to environmentally
period, suggesting the beginning of recovery of the community.
relevant concentrations of nickel
During the RPD period, Rotifera showed the best recovery, followed by Copepoda
C Pinheiro1; S Lopes1; A.M.V.M Soares1; S Loureiro1
and Branchiopoda. Plus, in the RPD period, some fluctuations in both abundance
and species diversity were observed. These fluctuations were also observed in the
The cladoceran Daphnia magna Straus was exposed to two environmentally
control, suggesting that the communities exposed to different salt concentrations
relevant concentrations of nickel (25 and 50 µg/L) for 12 successive
were returning to an initial equilibrium state, being a possible indication that the
generations to evaluate development of tolerance. Neonates from each
zooplankton community can recover after exposure to a seawater pulse.
generation pre-exposed to nickel or to control conditions were again exposed
Overall, the results showed a decreased abundance and richness of communities
to a concentration range of nickel to assess effects on life-cycle parameters
after a pulse of seawater intrusion, suggesting that saline stress may either inhibit or
(survival and feeding) . Significant changes in 48h-LC 50 values for survival
unstabilise the changes that could naturally occur in lenthic ecosystems. However, the
were observed after 8 generations. D. magna pre-exposed to 25 µg/L of nickel
increments in abundance of the macroinvertebrate and zooplankton communities
were more sensitive than control D. magna after 8-9 generations, becoming
during the PD phase, suggest that invertebrates are capable of recovering from salt
more tolerant to nickel reaching generation 10. After 11-12 generations
stress.
a significant increase in sensitivity to nickel was observed again. D. magna
pre-exposed to 50 µg/L of nickel were more sensitive than control D. magna
after 9-10 generations, but in the last 2 generations a significant increase in
1CESAM/Department of Biology, University of Aveiro;
2Centre for Functional Ecology, Department of Life Sciences, University of Coimbra/Escuela de Ciencias
Agrícolas y Ambientales, Pontificia Universidad Católica del Ecuador;
3Centre for Functional Ecology, Department of Life Sciences, University of Coimbra/Instituto Politécnico
de Coimbra, Escola Superior Agrária, Bencanta, Coimbra;
4Centre for Functional Ecology, Department of Life Sciences, University of Coimbra
tolerance to nickel was found. Although not always significant, 24h-EC 50 values
for feeding activity of D. magna pre-exposed to 50 µg/L of nickel were about
1.3-2.7 higher than those observed for control D. magna in all generations,
suggesting tolerance to nickel. Significant increases in tolerance to nickel
for feeding activity of D. magna pre-exposed to 25 µg/L of nickel were only
registered after 2, 5-6 and 10-11 generations. As nickel tolerance apparently
varies with successive generations, it may be concluded that multigeneration
exposure studies are important for the evaluation of the long-term effects
77
of environmental toxicants. This study also demonstrates that organisms’
Aquatic Toxicology
capacity to develop tolerance is essential to ensure the accuracy of ecological
risk assessments.
Antioxidant responses and bioaccumulation in clam Scrobicularia plana
exposed to arsenate
1Department of Biology & CESAM
C. Trombini1; M. Hampel2; G. Rodríguez-Moro3; T. García-Barrera3; J.L. Gómez-Ariza3;
J. Blasco1
Arsenic is a well known toxicant released into the environment through different
industrial processes and agricultural usage which environmental concentrations
normally between 1 and 10 µg/L reaching milligram concentrations in polluted areas.
In the aquatic environment arsenic exists in different chemical forms being arsenate
(Asv) is the most common form under oxic conditions. In the present study marine clam
Scrobicularia plana was used as a model organism to investigate oxidative stress and
bioaccumulation related to Asv exposure. Clams were exposed to two concentrations
of Asv (10 and 100 µgL-1) for 14 days and samples were collected after a 1, 7 and
14 days of exposure. Antioxidant biomarker including glutathione S-transferase
(GST), glutathione reductase (GR), glutathione peroxidase (GPX), aceticholinesterase
(AchE) and superoxide dismutase (SOD) as well as lipid peroxidation (LPO) in gills
and digestive glands were analyzed. .SOD activity increased in both tissues after one
day of exposure to the highest concentration of Asv. A significant decrease in AchE
and GR activities was observed in gills after seven days at both concentrations. Also
GR activity decreased in digestive gland at the day 14. On the other hand, GST, GX
and LPO didn’t show any significant alteration over time. The results indicate that
exposure to environmental concentrations of Asv can induce oxidative stress. The
concentration of Asv was determined in clam digestive gland: S. Plana showed a
moderate bioaccumulation ability when exposed to both concentrations of Asv.
79
1Instituto de Ciencias Marinas de Andalucía (CSIC), Campus Rio San Pedro, 11510 Puerto Real, Cádiz, Spain;
2Centro Andaluz de Ciencias y Tecnologías Marinas (CACYTMAR), Universidad de Cádiz, Campus
Universitario de Puerto Real, 11510 Puerto Real, Cádiz, Spain;
3Departamento de Química y Ciencia de los Materiales “Profesor José Carlos Vílchez Martín”. Campus del
Carmen. Av. Fuerzas Armadas s7n. 21071 Huelva , Spain
Aquatic Toxicology
Influence of temperature on the toxicity of cadmium and benzo(a)anthracene to planktonic microalgae
L. R. Vieira1; F. Morgado2; L. Guilhermino1
In the present context of global warming, more knowledge on the combined
effects induced by temperature changes and mixtures of relevant environmental
contaminants is urgently needed. Therefore, the main objective of this study was to
investigate the effects of temperature increase on the toxicity of a relevant metal and
polycyclic aromatic hydrocarbon (cadmium and benzo(a)anthracene), alone and in
mixture, to the marine planktonic algae Tetraselmis chuii. Microalgae cultures were
exposed for 96h to different concentrations of both substances individually and in
binary mixtures, at 20 and 25°C. The effect criterion was the inhibition of culture
growth. The increase of temperature by 5°C significantly increased the toxicity of both
tested contaminants at concentrations between 0.313 and 1.25 mgL-1. Toxicological
interactions among the stressors were found, highlighting the need of more research
on the topic.
1University of Porto: ICBAS & CIIMAR: ICBAS-Department of Population studies, Laboratory of Ecotoxicology;
CIIMAR-Research Group of Ecotoxicology, Stress Ecology and Environmental Health, Rua dos Bragas, 289,
4050-123 Porto, Portugal.;
2University of Aveiro: Department of Biology & CESAM, 3810-193 Aveiro, Portugal.
81
Aquatic Toxicology
Do cationic and hydrophobic modifications influence the ecotoxicity of
hydroxyethyl cellulose polymers?
A.M. Simões1; F.E. Antunes2; I. Lopes1
New highly charged polymers have been developed within the industry of
personal care products, aiming at better performances for conditioning and
deposit beneficial ingredients onto hair and skin. Among these, SoftCatTM
have proved to be more efficient than conventional cationic polymers.
They are polymers based on quaternary ammonium salts of hydroxyethyl
cellulose and include several families of cationic products that differ in both
viscosity and charge. SoftCAT SK comprise four high viscosity polymers that
incorporate variations in charge level and hydrophobic modification while
SoftCAT TM SL constitute four high viscosity polymers that incorporate low
levels of hydrophobic modifications. It has been shown that these variations
in architecture correspond to different performances. In the present work is
hypothesised that such differences in architecture are also associated with
different ecotoxicological effects. If such association is detected the investment
on the less toxic variation could be recommend to the industry. To test above
hypothesis, the following standard toxicity assays were carried out for each
variation of SofCats TM: bioluminescence inhibition with the bacteria Vibrio fisheri
and the 72h-growth inhibition with the freshwater microalgae Raphidocelis
subcapitata. Preliminary results suggest that the variations of SoftCAT SL
showed the highest toxicity in relation to SoftCAT SK. However, further studies,
with more complex organisms, are still needed to fully understand the adverse
83
ecological effects of these type of polymers and allow suggesting which
Aquatic Toxicology
comprise the more environmental friendly solution simultaneously retaining
the desired functions.
A “stock culture independent” duckweed microbiotest with Spirodela
polyrhiza: a practical and low cost alternative to Lemna bioassays
1Department of Biology, CESAM, University of Aveiro, Aveiro; Portugal;
2Department of Chemistry, University of Coimbra, Coimbra, Portugal
Persoone Guido1; R.Baudo2; M.Foudoulakis3; G.Arapis4
Toxicity tests with duckweeds are performed for already many years in aquatic
toxicology, and standard tests procedures with Lemna species have been published
by national and international organizations. In Europe, duckweed tests are mandatory
for risk assessment of herbicides and plant growth regulators. Like virtually all
aquatic toxicity tests, bioassays with Lemna are dependent on stock culturing of the
test organisms. The assays require substantial materials, bench space and incubation
space. In order to bypass the former handicaps, a practical and low cost duckweed
microbiotest has been developed for performance in small multiwells, and which
makes use of the “dormant” vegetative stages (turions) of the duckweed Spirodela
polyrhiza which can be stored for long periods of time and germinated at the time
of performance of the assays. The 3 days Spirodela polyrhiza microbiotest is based
on the growth of the first frond of the germinated turions as the effect parameter.
The areas are measured by Image Analysis on 2 photos of the multiwells, taken
at the start (= t0h) and at the end (t=72h) of the toxicity test. The precision of the
Spirodela polyrhiza microbiotest has been determined in 3 extensive international
ringtests, the reports of which can be found on the website www.microbiotests.be.
A sensitivity comparison of the EC50’s of the 7 days Lemna minor assay and on the
3 days new microbiotest has been made on 22 chemicals, including herbicides and
metals. The high R2 value (0.98) calculated from the comparison of the data pairs
showed that the Spirodela polyrhiza test has the same sensitivity as he conventional
85
Lemna test. A proposal has been submitted to the ISO for accepting the Spirodela
Endocrine Disruption
polyrhiza duckweed microbiotest as a new standard toxicity test for wastewaters,
natural waters and chemicals.
Effects of the endocrine disruptor vinclozolin in the benthic organism
Chironomus riparius (Diptera).
1Ghent University/MicroBioTests Inc.;
2CNR – Istituto per lo Studio degli Ecosistemi, Verbania, Pallanza, Italy;
3Dow AgroSciences, Lavrion, Greece;
4Laboratory of Ecology and Environmental Sciences, Agricultural University of Athens, Athens, Greece
Mónica Aquilino Amez1
The Vinclozolin (Vz) is a widely used pesticide for agricultural purposes, its use and
presence in the environment has increased significantly in recent years, especially in aquatic
ecosystems. It has been described as an endocrine disruptor in vertebrates showing an
antiandrogen behavior. However, despite being an important part of the food chain,
studies with invertebrates are scarce so the mode of action of Vz and its interaction with the
endocrine systems are unknown. In this work we have analyzed the molecular effects caused
by Vinclozolin in fourth instar larvae and embryos of Chironomus riparius (Diptera), a benthic
organism of inland water. After exposing the larvae and embryos to this compound for 24
hours, changes in expression of three genes were analyzed by Real-Time PCR. Two of them,
the ecdysone receptor (EcR) and E74, are related to the ecdysone response pathway, which is
related with development in insects, while the third, Hsp70, has a role in cell stress response.
The results show a significant alteration in expression levels of EcR and Hsp70 genes, indicating
that different concentrations of Vz affect these cellular processes at different stages of the life
cycle of Chironomus riparius. This is the first evidence at molecular level of the mode of action of
Vinclozolin on the endocrine system in invertebrates, which highlights the need of additional
research to elucidate the effects of this compound in a key organism of aquatic ecosystems.
This work was supported by the Plan Nacional de Investigación Científica, Desarrollo e Innovación
Tecnológica (Spain), grant CTM2012-37547 from the Ciencias y Tecnologías Medioambientales program.
M.A. is the receiver of a predoctoral contract Ministry of Economy and Finance (BES-2013-064041).
1Universidad Nacional de Educación a Distancia.
87
Endocrine Disruption
Developmental effects of a xenoestrogen and an aromatase-inhibitor in
zebrafish (Danio rerio) embryogenesis
D. Santos1; M. Matos2; A.M. Coimbra3
The pollution of aquatic ecosystems with endocrine-disrupting chemicals (EDCs)
is increasingly drawing public attention. EDCs, which include many natural and
synthetic compounds, are capable of interfering with the endocrine system functions
through multiple mechanisms of action. In fact, the exposure to EDCs has been linked
to disruption of the sexual development and differentiation in fish. However, the
effects of EDCs on the full embryonic development have received limited attention.
In the present study, zebrafish (Danio rerio) embryos and larvae were used
to assess how EDCs may interfere with early life development. For this purpose,
zebrafish embryos were exposed to nominal concentrations of the xenoestrogen
genistein (Gen: 2, 20, 200 and 2000 ng/L) and of the aromatase inhibitor fadrozole
(Fad: 2, 10, 50 and 250 μg/L), between 2 and 144 h post-fertilization (hpf ). Several
endpoints, namely, somite development, heartbeat, malformations, mortality and
hatching rates were evaluated. Additionally, the expression levels of hormone
receptors (esr1, esr2a, esr2b and ar) and apoptotic pathways related genes (p53
and c-jun) were determined using quantitative real-time PCR (QPCR). Our results
showed that Gen and Fad exposures led to increased mortality and malformations
in zebrafish embryos/larvae. Somites development was not affected by both
compounds, while a significant effect was observed in the heartbeat rate, at 144 hpf,
in larvae exposed to Fad. QPCR revealed alterations in the expression levels of all the
studied genes, at different time points. Gen up-regulated the esr1 and c-jun genes,
89
while the expression of esr2a, esr2b and ar genes was down-regulated. Fad exposure
Endocrine Disruption
decreased esr1, p53 and c-jun expression levels.
Our results showed that xenoestrogens and/or aromatase inhibitors, such as Gen
and Fad, have the potential to induce negative impacts on zebrafish embryogenesis.
Differences in the xenoestrogenic sensitivity of five teleost species exposed
to 17α-ethinylestradiol
Andere Basterretxea1; Gorka Lorenzo1; Cristina Bizarro1; Eider Bilbao1;
Maren Ortiz-Zarragoitia1
This work was supported by the Portuguese Foundation for Science and Technology (FCT) through the
project grant: PTCD/CTV/102453/2008 (FCOMP-01-0124, FEDER-009527)
1Life Sciences and Environment School, University of Trás-os-Montes and Alto Douro (UTAD), Quinta de
Prados 5000-801 Vila Real, Portugal;
2Center of Agricultural Genomics and Biotechnology (CGBA), Department of Genetics and Biotechnology,
University of Trás-os-Montes e Alto Douro (UTAD), Quinta dos Prados, 5001-801 Vila Real, Portugal;
3Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University
of Trás-os-Montes and Alto Douro (UTAD), Quinta de Prados 5000-801 Vila Real, Portugal
Exposure to estrogenic chemicals, or xenoestrogens, discharged into the aquatic
environment has been shown to induce changes in hormonal homeostasis and
transcription levels of reproduction-related genes (i.e. vtg and cyp19a1b) in fish,
including the possibility of compromising the viability of affected populations
altering the normal gametogenesis and development. Xenoestrogenic responses
in fish vary according to the environment (freshwater vs marine), temperature and
species sensitivity. Thus, extrapolation of data between species inhabiting different
habitats and/or showing different behavioral habits within a given ecosystem could
result in erroneous effect estimation. The aim of the present work was to study the
sensitivity to 17α-ethinylestradiol (EE2) exposure in five fish species, including two
freshwater species (zebrafish and rainbow trout) and three marine species (European
sea bass, turbot and thicklip grey mullet). For this purpose well known biomarkers of
xenoestrogenicity in fish, such as transcription levels of vitellogenin (vtg), choriogenin
(chg) and brain aromatase cyp19a1b, were quantified by qRT-PCR. Juvenile fish were
exposed for 10 days to 5 ng/L, 25 ng/L and 50 ng/L of EE2 and samples of liver and
brain collected after 2 and 10 days of exposure. Results showed increased values
of hepatosomatic somatic index (HSI), suggesting changes on hepatic metabolic
activity. Accordingly, up-regulation of vtg was detected in studied species, being vtg
transcription more pronounced than cyp19a1b in all species except for thicklip grey
mullets. Comparing species sensitivity, the two studied freshwater species (zebrafish
91
and rainbow trout) were more sensitive to EE2 than the marine species. This work
Endocrine Disruption
demonstrates the species specific responses to xenoestrogens and highlights the
importance of characterizing the sensitivity to endocrine disrupting chemicals of
Gonadal development and biological recovery of zebrafish exposed to
selected species.
17ß-estradiol (e2) and 17α-ethinylestradiol (ee2)
Juliana Polloni Silva1; Gilberto Dias de Alkimin1; Samyra Chang Nakahira1; Cristina Elena de Souza
Correa1; Renata Fracácio1
Acknowledgements
Basque Government (S-PE13UN131-TOXADEN and Consolidated research groups IT-810-13); UPV/EHU
(UFI11/37) and Spanish Ministry of Economy and Competitiveness (AGL2012-33477-SEXOVUM).
Studies on toxic compounds not yet considered in the legislation, such as
endocrine disruptors (EDs), are of great scientific interest. We evaluated the toxicity of
1Plentzia Marine Station, University of the Basque Country UPV/EHU
17ß-estradiol (E2) and 17α-ethinylestradiol (EE2) hormones on adult female zebrafish
(Danio rerio) during chronic tests in which the water was contaminated in laboratory
using a concentration found on nature (30 ng.L-1, nominal concentration). Then,
we evaluated three types of materials (powdered activated carbon, PAC; aquatic
humic substances, AHS; and zeolites, ZEO) in an attempt to remove these hormones
from water, and we tested the efficiency of each substance through performing
biological recovery experiments with the same organisms. 38 fishes were exposed
to each experimental condition (control, E2 and EE2) during 21 days. At the 21th
day, randomly selected organisms (n=3) were euthanized for gonads removal. The
remaining fishes were exposed to recovery tests for more 7 days with test solutions
shaked for two hours with PAC (0,5g.L-1, n=12), ZEO (0,5g.L-1, n=12), and SHA (20 mg
L-1, n=11). At the 28th test day, the same number of organisms (n=3) were selected for
gonads extraction. Gonads were submitted to procedures of tissue fixation (Bouin
liquid over 18 hours), washing (running water over 12 hours), dehydration (with 70%
alcohol) and paraffin included. Serial sections (4μm) were obtained by microtome
and stained with hematoxylin-eosin (HE). Qualitative analyses were conducted in a
light microscopy and the histological anomalies classified according to OECD (2010).
The analyzes showed predominance of atresic follicles in all the controls, indicating
93
spawning and proper functioning of this tissue. The exposure to E2 did not result in
Endocrine Disruption
histological changes that could compromise the reproductive aspects of the species,
nevertheless, fishes exposed to EE2 showed predominance of initial maturation
Evaluation of zebrafish gonad development after endocrine disruptors’
stages indicating a delay on reproductive capacity, which remained irreversible after
exposures
biological recovery tests with all treatments and controls.
Ana Luzio1; Sandra M. Monteiro1; Sofia Garcia-Santos1; Eduardo Rocha2; António FontaínhasFernandes1; Ana M. Coimbra1
1São Paulo State University (UNESP - Campus of Sorocaba)
Zebrafish (Danio rerio) has long been used as a model in toxicological studies,
regarding the effects of different classes of environmental contaminants, such
as endocrine disruptor compounds (EDCs). Current knowledge on zebrafish sex
determination suggests that this trait has a polygenic genetic basis, although
environmental factors, such as EDC’s, may also be involved and perturb both sex
differentiation and development.
This study aimed to assess how sex steroids imbalance impacts sex differentiation
and gonad development in zebrafish. Fish where exposed to an estrogen
(17α-ethinylestradiol, 4 ng/L), to an inhibitor of estrogen synthesis (Fadrozole, 50 μg/L)
or to their binary mixture (17α-ethinylestradiol+ Fadrozole, 4 ng/L+50 μg/L), from 2
hour to 60 days post-fertilization (dpf ). Through a stereological approach, both relative
and absolute volumes of zebrafish gonad compartments were determined at 60 dpf
to identify alterations on gonad differentiation and development. The stereological
analysis suggested thatthe 17α-ethinylestradiol stimulus enhanced both zebrafish
body growth and gonad development. On the other hand,exposure to Fadrozole
affected the sexual development in zebrafish, inducing 90% masculinization, with
some degree of intersex being observed in males. The mixture seemed not to
affect the individual effects of each compound, with a masculinization index similar
to the one observed after Fadrozole exposed fish (90%), and females showing a
body weight gain as after 17α-ethinylestradiol exposure. Accordingly, the binary
95
mixture allowed the identification of sex dependent roles of steroid hormones, with
Biomarkers
estrogens being essential for ovary development and acting as growth promoters.
While, the potential increase in androgen levels in zebrafish induced masculinization
Ecotoxicological and biochemical effects of an herbicide and a metal on
that the addition of the estrogenic compound did not revert. Clearly, studying the
zooplankton and phytoplankton estuarine and marine species
effects of single EDCs may not be sufficient to understand what is really occurring in
Filimonova V.1; Gonçalves F.2; Marques J.C.3; De Troch M.4; Gonçalves A.M.M.5
the environment.
In Europe, mainly in the Mediterranean region, an intensive usage of pesticides
1Centre
for the Research and Technology of Agro-Environmental and Biological Sciences, CITAB,
Departamento de Biologia e Ambiente (DeBA), Escola de Ciências da Vida e Ambiente (ECVA), University
of Trás-os-Montes and Alto Douro, UTAD, Quinta de Prados, 500;
2Laboratory of Histology and Embryology, Institute of Biomedical Sciences Abel Salazar (ICBAS), University
of Porto (U. Porto), Portugal | Laboratory of Cellular, Molecular and Analytical Studies, Interdisciplinary
Centre of Marine and Environmental Resear
was recorded during the past 30 years. According to information from agricultural
cooperatives of the Mondego valley (Figueira da Foz, Portugal), Primextra® Gold TZ is
the most used herbicide in corn crops fields and one of the 20 best-selling herbicides
in Portugal. Copper is mainly used in pesticides’ formulation.This study aims to
determine the ecotoxicological and biochemical (namely fatty acids profiles) effects
of the herbicide Primextra® Gold TZ and the metal Copper. The study organisms are
three planktonic species: the marine diatom Thalassiosira weissflogii, the estuarine
copepod Acartia tonsa and the marine brine shrimp nauplii Artemia franciscana.
Fatty acids (FAs) are one of the most important molecules transferred across the
plant-animal interface in aquatic food webs, used as good biomarker of stress. In
our lab experiments, T. weissflogii showed to be the most sensitive followed by A.
tonsa to the herbicide (EC50=0.0078 mg/L and EC50=0.925 mg/L, respectively)
whereas the copepod was the most sensitive species to the metal followed by
T.weissflogii (EC50=0.234 mg/L and EC50= 0.383 mg/L respectively). A. franciscana
was the most tolerant organism to the herbicide and the metal (EC50=20.35 mg/L
and EC50=18.93 mg/L, respectively). Changes in the FA profiles of diatom and shrimp
nauplii were observed, with increasing saturation and decreasing unsaturation levels
and especially in highly unsaturated FAs, which are essential for zooplankton and
are mainly obtained from their food source. The same pattern was observed for A.
97
tonsa exposed to the herbicide. The results of this work suggest that discharges of
Biomarkers
Primextra or other pesticides mainly composed of copper damage the planktonic
populations and their biochemical composition, which may also cause changes in
The influence of different microalgae diets on cell and tissue level bio-
their nutritive value, with severe repercussions for higher trophic levels and thus to
markers in mussel digestive gland
the entire food web.
E. Blanco-Rayón1; M. Ücker1; L. Garmendia1; I. Marigómez1; U. Izagirre1
1IMAR-CMA
& MARE, Faculty of Science and Technology,University of Coimbra, 3004-517 Coimbra,
Portugal; Department of Biology & CESAM, University of Aveiro, 3810-193 Aveiro, Portugal; Ghent
University, Biology Department, Marine Biology Section, Krijgslaan;
2Department of Biology & CESAM, University of Aveiro, 3810-193 Aveiro, Portugal;
3IMAR-CMA & MARE, Faculty of Science and Technology,University of Coimbra, 3004-517 Coimbra,
Portugal;
4Ghent University, Biology Department, Marine Biology Section, Krijgslaan 281-S8, B-9000 Gent, Belgium;
5IMAR-CMA & MARE, Faculty of Science and Technology,University of Coimbra, 3004-517 Coimbra,
Portugal; Department of Biology & CESAM, University of Aveiro, 3810-193 Aveiro, Portugal
Digestive gland of mussels is a target tissue for the assessment of the marine
ecosystem health status in monitoring programs as well as to investigate the biological
effects of pollutants in the laboratory. Standard procedures have been developed
for the determination of biomarkers; however, guidelines dealing with procedures
to maintain mussels during experimentation are lacking, and consequently feeding
strategies are disparate amongst studies. Taking into account the high plasticity of
mussel digestive gland, food composition can be important confounding factors
in experimental studies of the biological responses of mussels to pollutants and
other environmental stressors. Thus, the present study is aimed at investigating the
influence of different diets on cell and tissue level biomarkers in mussel digestive
gland. For this purpose, mussels (Mytilus galloprovincialis) were maintained under
laboratory conditions and fed using 4 different microalgae diets ad libitum for 1
week: (a) Isochrysis galbana; (b) Tetraselmis chuii; (c) I.galbana and T.chuii microalgae
mixture; and (d) commercial food (Shellfish Diet® microalgae blend, Acuinuga).
Lysosomal structural changes, intracellular accumulation of neutral lipids, and cell
type composition and structural changes of digestive alveoli were determined in
digestive gland. The digestive cell lysosomes were larger in mussels fed microalgae
mixture than in the rest of the experimental groups. Significantly higher intracellular
neutral lipid accumulation was observed in mussels fed I. galbana than in mussels
fed commercial food. Moreover, thicker digestive gland epithelium was recorded in
99
mussels fed I. galbana than in mussels fed T. chuii. Connective-to-digestive-tissue
Biomarkers
ratio was higher in mussels fed commercial food than in the rest of the experimental
groups. In general, the present results demonstrate that biomarkers are influenced by
Accumulation kinetics of copper and silver and assessment of the effects
the type of diet employed during experimentation. Therefore, standard experimental
exerted after dietary exposure in oysters Crassostrea gigas.
procedures, including consensus food composition, are required in order to achieve
A. Rementeria1; M. Mikolaczyk2; E. Blanco-Rayón1; L. Velasco1; U. Izagirre1; B. Zaldibar1; J. Schäfer2
reliable and comparable investigations on the biological effects of pollutants and
their applications for the assessment of marine ecosystem health status.
Oysters have been widely used as sentinel organisms in environmental water
health status programs because of their sedentary way of life and ability to accumulate
Acknowledgements
This work has been funded by the Ministry of Economy and Competitiveness (MINECO 2013-2015), and
by the Government of the Basque Country through a pre-doc grant to Blanco-Rayón (PRE_2013_1_640)
1Cell Biology and Environmental Toxicology Research Group; Research Center for Experimental Marine
Biology and Biotechnology (PiE-UPV/EHU), University of Basque Country, Areatza, Plentzia – Bizkaia,
Basque Country, Spain
pollutants with little metabolic transformation. For example, the French MusselWatch Program (RNO/ROCCH) has collected oysters for over 30 years in order to
assess coastal water quality levels. In this context, metal accumulation measurements
together with cell and tissue level biomarkers provide reliable information about both
environmental and oyster’s health status. Classical ecotoxicological approaches are
often based on single compound exposure and they do not usually consider possible
interactions between pollutants that may occur in the real environment. Monitoring
programs carried out in the Bay of Biscay have indicated that in several estuaries high
concentrations of copper and silver are present in water, sediments and biota. It is
well known that silver and copper provoke deleterious effects in the environment
and even in human health. Besides, a possible synergistic effect between copper
and silver accumulation has been recently described in oysters. Presently in order to
determine this synergistic effect, oysters Crassostrea gigas have been exposed for 21
days to a range of environmentally relevant concentrations of isotopically labelled
63Cu and 107Ag alone and in mixture through food pathway. Animals were fed for
20h with previously contaminated microalgae Isochrysis galbana exposed for 24h
to: 500 ng 107Ag/L; 8600 ng 63Cu/L and 500 ng 107Ag/L+8600 ng 63Cu/L. Preliminary
results indicated a successful experimental set up since oysters in general presented
101
an increased Condition Index, relatively low mortality and increased neutral lipids
Biomarkers
in connective tissue. On the other hand, significant differences in lipofuscine levels
were observed through the experiment. Metal accumulation kinetics are discussed
Sub-lethal effects of exposure to atrazine in gill cells of sea lamprey
and compared to those obtained from similar experiments with direct exposure of
downstream migrants
oysters to Cu and Ag isotopes in seawater.
M. J. Lança1; M. Machado1; A.F. Ferreira2; M. Candeias3; R. Ferreira3; I. Alves-Pereira3; J.S. Carrola4;
B.R. Quintella5; P.R. Almeida6
Acknoledgements
Work funded by Basque Government (GIC07/26-IT-393-07) and research project CTM2012-40203-C02-01MAR (MINECO).
In Portugal, atrazine (ATZ) was one of the most widely used herbicides in
agriculture, and despite prohibited since 2007, it still continued to be detected
in the surface and ground waters. Recent research demonstrated that the
1Cell
Biology & Environmental Toxicology Research Group, Research Centre for Experimental Marine
Biology & Biotechnology (PIE) & Zoology & Animal Cell Biology Department (Faculty of Science &
Technology), University of the Basque Country;
2Université de Bordeaux, UMR 5805 EPOC, Allée Geoffroy St. Hilaire, 33615 Pessac cedex, France
conditions experienced by anadromous fishes, while in freshwater, may be
critical to their subsequent fitness and survival in the sea. Several studies
established a decline in wild populations, most probably caused by a reduction
in recruitment related with pollution. In the case of sea lamprey (Petromyzon
marinus) trophic migration to the ocean, expose fish to several stress factors,
including chemical stress. Thus, it is crucial to assess the effects of sub-lethal
exposure to ATZ and predict potential consequences to juveniles of sea lamprey.
Fish were exposed to 50 µg L-1 and 100 µg L-1 of ATZ for 30 days and subject
also to a gradual increase in salinity up to 35. We analyzed gill histopathological
biomarkers, characterized the lipid profile of the basolateral membrane
(BLM) of gill cells, and determined NKA activity, to assess effects of sub-lethal
exposure to ATZ. Upon exposure to ATZ, there was a fatty acid saturation of
the BLM of gill cells and this correlated significantly (r=0.966) with the NKA
activity in the presence of the highest concentration of herbicide. Fish exposed
to 50 µg L-1 ATZ and salinity 35 showed at lamella epithelium level moderate
distal hyperplasia, rare fusion and low level of lifting. At 100 µg L-1 and salinity
35 presented thicker filaments with irregular structure, mild distal hyperplasia,
103
and also architectural and structural alterations of the epithelium, mild rupture
Biomarkers
of pillar cells and mild increased in chloride cells number inside the clusters.
These concentrations of ATZ lead to low number of gills lesions which doesn´t
Identification of molecular biomarkers of exposure to sanitary sewage
compromise the gill physiology and sea lamprey juveniles´ survival.
in oyster Crassostrea brasiliana
Tomas Bohn Pessatti1; Karim Hahn Luchmann2; Fabrício Flores Nunes1; Flávia Lucena Zacchi1;
Afonso Celso Dias Bainy1; Jaco Joaquim Mattos1
1Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Departamento de Zootecnia,
Universidade de Évora, Largo dos Colegiais 2, 7004-516 Évora, Portugal.;
2MARE – Centro de Ciências do Mar e do Ambiente, Faculdade de Ciências da Universidade de Lisboa,
Campo Grande, 1749-016 Lisboa, Portugal.;
3Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Departamento de Química,
Universidade de Évora, Largo dos Colegiais 2, 7004-516 Évora, Portugal.;
4Departamento de Engenharia Biológica e Ambiental, ECVA - Escola de Ciências da Vida e Ambiente,
Universidade de Trás-os-Montes e Alto Douro, Quinta dos Prados, 5001-801 Vila Real, Portugal;
5MARE – Centro de Ciências do Mar e do Ambiente, Departamento de Biologia Animal, Faculdade de
Ciências da Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal;
6MARE – Centro de Ciências do Mar e do Ambiente, Faculdade de Ciências da Universidade de Lisboa,
Campo Grande, 1749-016 Lisboa, Portugal, Departamento de Biologia, Escola de Ciências e Tecnologia,
Universidade de Évora, Largo dos Colegiais 2, 7004-516
Untreated sanitary sewage discharged into the aquatic environment, together
with inadequate treatment plants is quite common in developing countries,
including some coastal regions of Brazil. Sewage is a complex mixture of toxic
compounds which may directly affect exposed organisms. Only chemical analyzes
do not reveal the actual risk of exposure to contaminants in biota. More sophisticated
approaches are required to better elucidate this information, such as the use of
molecular biomarkers. The present study evaluated the transcript levels of xenobiotic
biotransformation genes in gills of Crassostrea brasiliana kept for 24 hours in five sites
with different levels of contamination by sewage in Florianopolis, southern Brazil.
The transcription of five cytochrome P450 genes (CYP2AU1, CYP20A1-like CYP3A29like CYP356A1-like and CYP20A1-like), four glutathione S-transferase gene (GST-like,
GSTpi-like, GSTomega-like and GSTmicrosomal3-like) and a sulfotransferase (SULT-like)
were analyzed by quantitative reverse transcription PCR (qRT-PCR). The animals kept
in the contaminated site(Bucheler River) showed higher levels of transcription of
CYP2AU1 and SULT-like (approximately 20 and 50 times, respectively), when compared
to the reference group (Ratones Pequeno Island). The results are in agreement with
previous studies which reported the induction of CYP genes in Crassostrea gigas
kept in the same sites. Bucheler river has been previously characterized as heavily
contaminated by sanitary sewage, through biological and chemical analysis.
Therefore, our results show that some molecular biomarkers analyzed in the oyster C.
105
brasiliana after short-term in situ exposure respond to the presence of contaminants
Biomarkers
and evidence its potential use as sentinel organism to monitor coastal regions in
Brazil. Supported by INCT-TA and CNPq.
Biomarkers, histopathology and condition indices in sole (Solea senegalensis) exposed to contaminated sediment
1Universidade Federal de Santa Catarina;
2Universidade do Estado de Santa Catarina
Tifanie Briaudeau1; Edurne Huerga1; Ionan Marigómez1; Larraitz Garmendia1; Joxe Mikel Garmendia2; Izaskun Zorita2; Urtzi Izagirre1; Urtzi Izagirre1
Sediments are a major sink of contaminants entering estuaries and coastal
systems. They represent a potential ecological risk as they can act as a source
of persistent inorganic and organic contaminants to the water column. Flatfish,
amongst which the sole (Solea sp.) is common in the Bay of Biscay, are recognised as
sentinel species in pollution monitoring programmes. In the present investigation
and with the aim to determine the suitability of Solea senegalensis as sentinel
species, juveniles(22.381.86cm standard length) were exposed to contaminated
sediments for 28 d. Sediments were taken from a reference estuary (Plentzia, Basque
Coast; SED1) and from a highly polluted harbor (Pasaia, Basque Coast; SED3); a 1:1
v/v mixture of both (SED2) was used as well. In parallel, a 10-day acute survival test
with marine amphipods was carried out; this indicated toxicity in SED2 and SED3.
Upon chemical analysis of the source sediments, the highest concentration of
PAHs was recorded in SED1, whilst the concentration of metals (e.g. Cd, Cr, Cu)was
highest in SED3. For each experimental group, samples (N=12) were retrieved at 0, 3,
7 and 28d for the assessment of biological responses to pollutants. Hepatosomatic
and condition indices were calculated and biomarkers and histopathological
alterations were determined in liver, kidney, gills and gonad. Overall, differences
have been observed between experimental groups. Histological lesions such as fat
vacuolisation, melanomacrophage centres and kidney tubule alterations have been
detected in target organs with higher prevalence in SED3. Lysosomal structural
107
changes and lipid accumulation showed differences among experimental groups.
Biomarkers
This study confirmed the suitability of S. senegalensis as sentinel species through
biomarkers and histopathological approaches.
Is there endocrine disruption in male mugilids from the Tagus Estuary?
J. Carrola1; S. Ribeiro2; P. Almeida R.3; S. Pedro4; E. Rocha5; J. Neto6; MJ. Pires1; C. Cruz6; F. Ribeiro4;
C. Levita6; J. Ferreira-Cardoso1; A. Fontainhas-Fernandes1
Acknowledgment
Work funded by Spanish MINECO (CTM2012-40203-C02-01 and PhD fellowship to T.B.), University of the
Basque Country– UPV/EHU (UFI 11/37) and Basque Government through Consolidated Research Groups
fellowship (IT810-B).
1Cell Biology in Environmental Toxicology Research Group (CBET), Research Centre for Experimental
Marine Biology and Biotechnology (PiE-UPV/EHU), University of the Basque Country, E-48620 PlentziaBizkaia, Basque Country, Spain;
2AZTI, Herrera Kaia, Portualdea z/g, 20110, Pasaia, Basque Country Spain
Numerous pollutants have the capacity to interfere with the endocrine system of
fish (endocrine disrupting compounds - EDC), and can cause diverse adverse effects.
One histological endpoint, a common biomarker of estrogenic exposure/effect, is
the “intersex condition” - ovotestis: presence of oocytes in the testis parenchyma.
The aim of this work was to assess the prevalence and severity of ovotestis in 41
grey mullets (Mugilidae), belonging to three species: Liza aurata (18), Chelon labrosus
(22) and Liza ramada (1). The fish were caught in November of 2014 (Seixal bay Tagus estuary), with 38.9±2.7 cm and 487.7±91.6 g for L. aurata, 39.3±3.7 cm and
659.9±188.6 g for C. labrosus, 37.8 cm and 450 g for L. ramada.
The histology showed that 22% of the grey mullets caught were males (9/41), and
in all the slides analyzed by light microscopy no single oocyte was observed. This
result reinforces the data obtained in a previous survey performed in February 2014
at same location (showing 2 very small immature oocytes).
These data reflect different exposure to EDCs in mullets from others Portuguese
estuaries, namely Mondego, Douro and Ave, with approximately 9,5%, 18,4% and
27,5% of ovotestis respectively and Ovotestis Severity Index (OSI) around 0.5 in
Mondego mullets (stage 1), 5,4 in Douro mullets (stage 1), and 7,4 in Ave mullets
(stage 2) (Carrola, 2011).
These results suggest either a no or a low level biological exposure of potential
EDCs present in the Tagus estuary, i.e., not reaching a point of being able to induce
ovotestis. This can be related with the presence of many waste water treatment plants
109
(WWTP) in the vicinity of the estuary, which are removing the suspended particulate
Biomarkers
matter (SPM) in the wastepipes, thus lessening (at least) the input/exposure by
ingestion. More studies are warrant, with more sensitive biomarkers (like hepatic
Background concentrations of polycyclic aromatic hydrocarbons me-
expression of vitellogenin), to definitely answer the question at stake.
tabolites in Portuguese firemen
M. Oliveira1; K. Slezakova2; A. Fernandes3; J.A. Vaz3; C. Delerue-Matos1; M.C. Pereira2; S. Morais1
Acknowledgements
Ana Silva, Margarida Raposo, Pedro Oliveira, Luis Cerdeira and Ana Fraga.
1Centre
for the Research and Technology for Agro-Environmental and Biological Sciences (CITAB),
University of Trás-os-Montes and Alto Douro (UTAD), Quinta de Prados 5000-801 Vila Real, Portugal;
2University of Trás-os-Montes and Alto Douro (UTAD), Vila Real, Portugal;
3MARE - Marine and Environmental Sciences Centre, Faculty of Sciences, University of Lisbon, Portugal;
Department of Biology, School of Sciences and Technology, University of Évora, Portugal;
4Department of Biology, School of Sciences and Technology, University of Évora, Portugal; 5Interdisciplinary
Centre of Marine and Environmental Research (CIIMAR), ICBAS, University of Porto UPorto, Portugal;
6Lusophone University of Humanities and Technologies, Lisbon, Portugal
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants
produced by the incomplete combustion of organic materials. PAHs may pose risks
to human health as many of the individual compounds are cytotoxic and mutagenic
to both lower and higher organisms, being some of them regarded as carcinogenic.
Pyrene is by far the most characterized PAH in all sample matrices, and is classified
as PAH marker of exposure while benzo(a)pyrene is considered the biomarker of
carcinogenic exposure to PAHs. Among the 16 PAHs established by US EPA as priority
pollutants, naphthalene, acenaphthene, fluorene, and phenanthrene are also found
in almost all the matrices.
Workers from industrial settings where airborne PAH levels are high such as coke
works and the primary aluminium industry, show excess rates of cancers. Firemen
are also exposed to high concentrations of PAHs during firefighting; however their
biomonitoring is difficult and epidemiological studies are scarce. During the last
decade, the urinary 1-hydroxypyrene has been used as a biomarker of environmental
and occupational exposure to PAHs. Still no standard reference or occupational
guidelines are available for any urinary PAH metabolite.
Within the present work, sixty healthy and no smoking Portuguese firemen
from ten Portuguese corporations from the district of Bragança (North of Portugal)
were evaluated regarding their levels of the most important urinary hydroxylPAHs:
1-hydroxynaphthalene,
1-hydroxyacenaphthene,
2-hydroxyfluorene,
111
1-hydroxyphenanthrene, 1-hydroxypyrene and 3-hydroxybenzo(a)pyrene. Firemen
Risk Assessment
were asked to fill a structured questionnaire to characterize the group and to identify
the potential exposure routes to PAHs.Hydroxyl-PAH concentrations were normalized
Wastewater reuse: a study of chloroform formation
with the respective urinary creatinine levels.
Anabela Rebelo1; Isabel Ferra2; Albertina Marques2; Isolina Gonçalves2; Rui Oliveira2; Margarida
Pereira3
Acknowledgements
This work was supported by the Fundação para a Ciência e a Tecnologia through the projects Pest-C/
EQB/LA0006/2013 and Pest-C/EQB/UI0511/2013. M. Oliveira and K. Slezakova are also grateful for their
fellowships SFRH/BD/80113/2011 and SFRH/BPD/65722/2009, respectively.
1REQUIMTE-LAQV, Instituto Superior de Engenharia do Instituto Politécnico do Porto;
2LEPABE, Departamento de Engenharia Química, Faculdade de Engenharia, Universidade do Porto; 3Escola
Superior de Saúde, Instituto Politécnico de Bragança
Wastewater reuse has been considered an appropriate and alternative water
source, e.g., for green areas irrigation. For safety reasons, recycled waters should
be chlorinated, to maintain a residual protection against microbiological regrowth.
However this disinfection procedure can lead to secondary reactions and subsequent
formation of halogenated compounds, such as trihalomethanes, that include
chloroform (CHCl3), a substance that presents a significant risk to or via the aquatic
environment, and named as a priority substance according to the European Water
Framework Directive.
Although the chloroform formation has been widely studied, the majority of
experiments have been carried out on natural organic matter (NOM) from surface
waters and less attention has been paid to wastewaters. When municipal wastewaters
are stored in landscape ponds (e.g., in golf courses), NOM from two distinct water
sources is present. Since the aromatic content of NOM depends on the type of
source, the disinfection by-products (DBP) formation is expected to follow a different
pattern.
The chloroform concentration and its correlation with chlorine dose and reaction
time in synthetic wastewaters for reclamation purposes were studied in this work.
Experiments were carried out in batch mode with a simulated wastewater for green
areas irrigation. A two variant empirical model is proposed to simulate breakpoint
chlorination practices, when chlorine dose is equal or lower than chlorine demand,
and super chlorination techniques, when chlorine dose tends to surpass chlorine
113
demand. Results show that the proposed model fits quite well the experimental
Risk Assessment
data and it was successfully applied to real data from wastewater treatment plants
This empirical model present a useful tool to be applied in reclaimed wastewaters to
Carbon capture and storage (ccs) strategy: a risk assessment overview
predict the range of CHCl3 formation and to adjust disinfection practices (chlorine
focused on marine bacteria
dose and contact time).
Ana Rocío Borrero Santiago1; Inmaculada Riba López1; Ángel Del Valls Casillas1
References
Deborde M, von Gunten U. Reactions of chlorine with inorganic and organic compounds during water
treatment-Kinetics and mechanisms: A critical review. Water Res. 2008; 42:13-51.
Espigares E, Moreno E, Fernández-Crehuet M, Jiménez E, Espigares M. Sustainable and effective control of
trihalomethanes in the breakpoint chlorination of wastewater effluents. Environ Technol. 2013; 34:231-7.
Koukouraki E, Diamadopoulos E. Modelling the formation of THM (trihalomethanes) during chlorination
of treated municipal wastewater. Water Sci Technol. 2003; 3:277-84.
Li S, Yang X, Qiu R, Wang P. Contents and leaching of trihalomethane precursors in soils. Water, Air, Soil
Pollut. 2003;145:35-52.
Xue S, Wang K, Zhao Q-L, Wei L-L. Chlorine reactivity and transformation of effluent dissolved organic
fractions during chlorination. Desalination. 2009; 249:63-71.
Xue S, Zhao Q, Ma X, Li F, Wang J, Wei L. Comparison of dissolved organic matter fractions in a secondary
effluent and a natural water. Environ Monit Assess. 2011; 180:371-83.
1Agência Portuguesa do Ambiente - ARH Algarve;
2Universidade da Beira Interior;
3Agência Portuguesa do Ambiente
Scientific community has focused on several aspects related to the ocean
acidification in the water column due to an increase of CO2 in the atmosphere,
such as pH variations, seawater chemistry and negative effects on diverse marine
organisms. Ocean acidification term has to be extended because a pH decrease
may also occur in deep water as a consequence of a CO2 leak from a stable
geological formation (carbon, capture and storage, CCS) or natural CO2 escapes.
CCS is one of the best options to storage carbon dioxide to mitigate the negative
effects of the climate change. However, this strategy may have associated some
risks such as CO2 leakages due to an escape from the reservoir. It is necessary
to improve the knowledge about the chemistry, nutrients fluxes, pollutants
mobilization and effects in benthic organisms. In this context, marine bacteria,
and all the process which they are involved, have been underestimated. In order
to figure out the gaps and the lack of knowledge, this work summarizes different
studies related to the potential effects on the marine bacteria related to an
acidification caused by CO2 leak fromCSS. Moreover, this contribution proposes
a strategy for laboratory protocols using Pseudomona stanieri (CECT7076) as
a case of study and analyzes the response of the strain under different CO2
conditions. Results showed significant differences in the growth rate under six
diluted enriched medium and differences about the days in the exponential
growth phase. Best mediums were selected to exposed P. stanieri under several
115
acidification conditions. The results of this work will provide an essential tool to
Risk Assessment
understand and develop a management strategy to improve future works related
to possible effects produced by potential CO2 leaks.
Maturity and stability parameters in the quality assessment of composts, sludges and other representative organic wastes intended to ag-
1Departamento de Química-Física, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz.
UNESCO/UNITWIN Wicop. Campus de Excelencia Internacional del Mar (CEIMAR). Polígono Río San Pedro
s/n, Puerto Real 11510 Cádiz, Spain
ricultural use
Paula Alvarenga1; Clarisse Mourinha1; Márcia Farto1; Patrícia Palma1; Joana Sengo2; Marie-Christine Morais2; Ana Cristina Cunha-Queda2
The quality of sludges, composts, and other representative organic wastes, with
potential to be used as agricultural soil amendments, was assessed using chemical
and biological parameters, typically used to evaluate the maturity and stability of
composts. The results evidenced that some organic materials, like agro industrial
sludge, municipal slaughterhouse sludge, and paper mill wastes, were identified as
“stable” by the tests based in the evaluation of microbial activity (e.g. Dewar selfheating and respiration activity AT4), only because they were chemically stabilized
with lime. However, these materials can have in their composition immature
organic matter and, as a consequence, once applied to soil can further proceed a
decomposition process. Germination tests were able to identify their phytotoxicity,
but, on the other hand, were not sensitive enough to identify the immaturity and
instability of some sewage sludges, likely because its organic matter was still in an
immature stage, containing low amount of salts and phytotoxic organic compounds.
Some enzymatic activities proved to be reliable tests to distinguish the organic
materials that were in an active stage of microbial activity, with high correlation
coefficients to NH4+-N content and NH4+-N/NO3--N ratio. In fact, these chemical
parameters evidenced as important in the quality assessment of an organic material,
strongly correlated with the biological parameters. The same was not true for the
majority of the humification indices, which proved inadequate to compare the
117
quality of diverse organic materials. Concluding, in a similar scenario, where the
Risk Assessment
organic materials in evaluation are dissimilar, both in the raw material an in the
state of the organic matter decomposition, the risk should be properly assessed by
Testing potential CO2 leakages on three marine microalgae: metal spe-
the integrated use of more than one parameter. From the results, we suggest the
ciation and biological effects
integrated use of the chemical parameters NH4+-N content and NH4+-N/NO3--N ratio,
Esther Bautista-Chamizo1; Manoela Romanó de Orte2; Inmaculada Riba López1; Ángel del Valls Casillas1
the biological parameters based in the evaluation of microbial activity (e.g. Dewar
self-heating, respiration activity AT4), one or two enzymatic acitivities (e.g. acid-
The greenhouse gas control strategy named CCS (carbon capture and storage) has
phosphatase, alkaline-phosphatase, β-glucosidase, protease or β-glucosaminidase),
been proposed as a mitigating process to current climate change. Ocean acidification has
and a germination test, to evaluate the phytotoxicity of the material.
been linked with this climate change and involves severe changes on the physicochemical
characteristics of the ocean and is associated with and unpredictable ecological risk.
1DTCA - Instituto Politécnico de Beja, Rua Pedro Soares S/N, Apartado 6155, 7800-295 Beja, Portugal;
2LEAF and Instituto Superior de Agronomia, Universidade de Lisboa, Tapada da Ajuda, 1349-017 Lisboa,
Portugal
While CCS pretends to soften the consequences of greenhouse gases, the viability of this
technology is still under study due to potential CO2 leakages from sub-seabed storage sites.
Apart from the ecological risk, metal speciation has been largely associated with pH and
redox condition. In this sense, growth-inhibition tests were developed with three marine
microalgae (Pleurochrysis roscoffensis, Cyclotella sp. and Andifinium sp.) being exposed to
five selected pH values (8, 7, 6.5, 6 and 5.5). Laboratory-scale experiments were carried out
during 10 days with the aim of testing metal bioavailability and microalgae growth. A full
inhibition of these marine microalgae was experimented at pH 5.5. In contrast, Pleurochrysis
roscoffensis showed an enhanced growth at pH 6, featuring a smaller sensitivity under
acidification conditions respect with the other two species, which start growing at pH 6.5.
The results obtained from this study with different species of marine microalgae are useful
to understand and evaluate the potential risk of this technology on the marine environment.
Keywords
CCS, marine microalgae, metal speciation, risk assessment.
1Universidad de Cádiz; 2Universidade Federal de São Paulo
119
Risk Assessment
Rational design for safer nanomaterials: a case study with nano-biosurfactants.
Nuno Martins1; Joana L. Pereira1; Duarte C.M.G2; Lídia C. A. Dias1; Antunes F.E.3; Isabel Lopes4
The increased production and consume of nanomaterials (NM) is boosting their
release to the environment, which highlights the need to assess their risk to and
ensure the protection of recipient ecosystems. With international policies pushing
for innovation in the field of green technologies, nanotechnology configures the
opportunity to avoid inefficiencies of the classic chemical industry. A major example
is the use of rational design principles to reduce the environmental hazardous
potential of new NMs. Within this scenario, cooperation between industry and
research in early product development stages may be the key to avoid investment
in NM variants that will not comply with environmental regulations. As part of such
a cooperation project, ecotoxicological trials were conducted for seven variants of
a soft organic NM made of biosurfactants; the variants were obtained following the
functionalization of the headgroup with an increasing number of ethylene oxide (EO)
units. Standard assays with freshwater species belonging to different taxonomic and
functional groups were performed: growth rate assay with the algae Raphidocelis
subcapitata, mortality and reproduction assay with the rotifer Brachionus caliciflorus,
growth rate with the ostracoda Heterocistis incongruens, mortality and embryo
development assay with the fish Danio rerio and the amphibian Pelophylax perezi.
Even though there was a slight tendency for the reduction of toxicity with the increase
of EO units, most species showed an overlap in confidence limits that prevented a
clear ranking of the variants. The seven variations of NM biosurfactants were found
121
toxic to freshwater biota. Tolerance to the NMs changed drastically depending on
Risk Assessment
the species, with R. subcapitata being the least sensitive species and B. calyciflorus
the most sensitive. Since the EO units functionalization only mildly affects the NMs
Determination of estuarine vulnerability to contamination for use in
toxicity, this structural feature can hardly be used alone as a safety variable in their
ecological risk assessment
rational design towards meeting a safer biosurfactant variant.
D.C. Ribeiro1; L. Guilhermino1
1Departamento de Biologia, CESAM, Universidade de Aveiro;
2Department of Chemistry , University of Coimbra , Coimbra
, Portugal; 3Department of Chemistry ,
University of Coimbra , Coimbra , Portugal;
4Department of Biology, CESAM, University of Aveiro, Aveiro, Portugal
A high number of estuaries are under strong anthropogenic pressures, including
contamination by hazardous chemicals from both continental (e.g. industrial and
urban discharges) and offshore sources (e.g. oil spills). The ecological risk assessment
(ERA) of these chemicals is crucial to support protective measures, management
and remediation actions. A fundamental step in the Problem Formulation of the ERA
process is an effective characterization of the ecosystem, including contamination
sources and system vulnerability. In the present study, a general framework to
assess the system vulnerability applying both inter- and intra-system analysis, and
based on the multi-metric index approach was developed. This framework delivers
information on the system’s vulnerability, identifies especially vulnerable zones of
the system (spatial discretization) and the most critical period(s) for contamination
events (temporal discretization). This information is of great importance as it provides
orientation of “where” and “when” the assessment efforts should be concentrated.
The framework was tested using the Minho River estuary as a case study, considering
the best and worst case scenarios for both continental and offshore (reaching the
coast) contamination events, and evaluating the ecosystem potential responses.
1University of Porto: ICBAS & CIIMAR: ICBAS-Department of Population studies, Laboratory of Ecotoxicology;
CIIMAR-Research Group of Ecotoxicology, Stress Ecology and Environmental Health, Rua dos Bragas, 289,
4050-123 Porto, Portugal.
123
Risk Assessment
Phosphorus fractionation in sediments from a small-sized dam in a rural
mountainous catchment: case study in NE Portugal
Marta Von Hafe Roboredo1; Reis, A.R.2; Pinto, J.P.M.1; Oliveira, A.I.1; Parker, A.3
Small reservoirs having less capacity to retain sediments are widely distributed in
regulated basins and, collectively, impart an important anthropogenic signature to
sediment’s quantity and quality. This study was carried out in the mini-hydropower
dam of Terragido (Vila Real - NE Portugal). The land use of the drained area is mainly
forest and agriculture with scattered urban settlements. Before the construction of
the WWTP, the urban effluents were discharged in the river network and the water
in the reservoir was of poor quality. Although a major improvement has been
achieved, sediments rich in Fe and Al oxides and hydroxides retaining particulate P
can be subject to transformations, altering its liability to release P and consequently
affecting the trophic state of the water body.
The inorganic P sequential fractionation (Chang and Jackson proposal) was
performed in bottom sediments (< 63µm), with the aim of understanding P mobility
and susceptibility to water transfer. The fractions considered were 1M NH4Cl (soluble
and loosely bound P), 0.5M NH4F (Al-P), 0.1M NaOH (Fe-P), CDB (reductant soluble P),
0.25M H2SO4 (Ca-P).
The sum of contents from all fractions varied between 1518 and 2454 mg P kg-1;
P was largely retained by Fe and Al oxide/hydroxides. Most samples revealed the
predominance of the Fe-P fraction (34% - 48%), nevertheless P extracted by the Al-P
fraction was lower (21% - 48%), followed by Ca-P (13% - 24%) and CDB (5% - 13%).
Soluble and loosely bound P was < 0.1% in all the samples. Higher P contents occur
125
at the river Corgo (2454 mg P kg-1), near the entrance of the reservoir, which could be
Risk Assessment
ascribed to the influence of anthropogenic activity, namely agriculture (Ca-P fraction
is also high). In the downstream part of the reservoir P contents show an increase
Environment risk assessment and bioaccumulation of metals in the met-
(1794 and 2081 mg P kg-1).
al contaminated basin
Estefania Bonnail1; AM. Sarmiento2; TA delValls1; I Riba1; JM Nieto3
1Department of Biology and Environment, University of Trás-os-Montes e Alto Douro (UTAD), Quinta de
Prados, 5000-801 Vila Real, Portugal;
2Department of Geology, University of Trás-os-Montes e Alto Douro (UTAD), Quinta de Prados, 5000-801
Vila Real, Portugal | CEMUC - Centre for Mechanical Engineering, University of Coimbra, Coimbra, Portugal;
3Soil Research Group, School of Human and Environmental Sciences, University of Reading, Reading, UK
The Iberian Pyrite Belt (Southwester Iberian Peninsula) is a sulphate massive unit widely
exploited by mining activities since Ancient times. The local geochemistry of soil composition
plus sulphide mine deposit residues located close to streams and rivers promote acid mine
drainage pollution all along the Odiel River, with an affection of more than 40% of the basin.
Bank sediments acts as sink of pollutants raising the toxicity of the ecosystems, becoming
an extreme polymetallic environment with life limited to extremophile organisms. These are
insufficient to carry an environmental risk assessment; therefore the Asian clam Corbicula
fluminea, was used as biomonitor tool under laboratory exposure to sediments to test the
metal bioavailability evolution in three different segments of the basin.
These segments correspond to three different degrees of contamination. They have
been hydro-chemical and sediment characterized related to metal bioaccumulation in soft
tissue of the clams exposed. Results showed positive correlations between sediments and
bioaccumulation. Through metal concentration in tissue monitoring is possible to determine
the contamination degree of the basin. Toxicity responses also maintained correspondence
to the toxic units found in sediment characterization through theoretical indexes.
1UNESCO UNITWIN/WiCop. Department of Physical-Chemistry. Faculty of Marine Sciences, University of
Cádiz. Campus Río San Pedro, 11510 Puerto Real, Cádiz. Spain;
2Department of Geodynamics and Paleontology, Faculty of Experimental Sciences, University of Huelva.
Campus ‘El Carmen’, 21071 Huelva, Spain;
3Department of Geology, Faculty of Experimental Sciences, University of Huelva. Campus ‘El Carmen’,
21071 Huelva, Spain
127
Global Changes
An ecological relevant approach to oil sands
Diogo N. Cardoso1; F. J. Wrona2; A.M.V.M. Soares1; Susana Loureiro1
The Athabasca oil sands deposits in northern Alberta, Canada, have
several special and specific characteristics that make them a special case
study, having both a high potential for hydrocarbon energy extraction and
also a region to assess associated environmental impacts. A key challenge
is understanding the sources, fate, distribution and effects of hydrocarbon
related contaminants in the ambient environment. An ecologically relevant
approach is to assess how organisms deal with inputs from natural processes
(erosion) or from human activities related releases of geological materials
from these areas to aquatic systems. Ecotoxicological tests were conducted
using parental geological material collected from the river banks near the
oil exploitation area. In a preliminary approach, the representative soil
invertebrate Folsomia candida was used to assess the effects of the parental
material on the reproduction output (ISO 22030 guideline (ISO 1999)). In a
complementary approach, aquatic bioassays were conducted with Daphnia
magna assessing the toxicological potential of erosional inputs into the
freshwater environment.
A range of elutriate extractions were obtained
to mimic a worst-case scenario of erosional substrate incorporation into
the aquatic systems. Three cycles of elutriate extraction were performed,
collecting the supernatant for Daphnia bioassays, and the pellet re-used for
a new cycle of extraction until a 3rd cycle was concluded. Preliminary results
showed that a suite of methodologies were needed to derive an accurate
129
assessment of effects. The use of both soil and aquatic organisms provided
Bioremediation
complementary knowledge on how these representative organisms react
to exposure of oil sands materials and to the potential erosion-related exposure
Synergistic and beneficial effects of 2,4-epibrassinolide and pro-
processes.
gesterone for the environmental remediation of progesterone by
Solanum nigrum L. Plants
1Department of Biology & Centre for Environmental and Marine Studies (CESAM), University of Aveiro,
3810-193 Aveiro, Portugal;
2Department of Geography, University of Victoria, 3800 Finnerty Road, David Turpin Building, Victoria, BC
V8P 5C2, Canada
A. Pinto1; C. Soares1; F. Fidalgo1; J. Teixeira1
In many sites around the world, steroid hormones’ levels have been reported to be
increasing, being designated as emerging pollutants. Progesterone (PRG), a member
of mammalian sex hormones, has been detected in various concentrations in sludge
samples from wastewater treatment plants, having potential adverse effects on
humans and wildlife’s endocrine systems, causing great concerns among the general
public and the scientific community. For these reasons, strategies for reducing the
accumulation of this steroid in the environment must be considered. In this work,
the phytoremediation potential of PRG by Solanum nigrum L. plants was evaluated.
For this purpose, 3 different treatments were performed: control (1); plants grown
in the presence of 0.768 µM PRG (2); and a third treatment, where plants grown in
the presence of 0.768 µM PRG were pretreated with a foliar application of 10 µM
2,4-epibrassinolide (EBL), to prevent a potentially stressful situation (3). In both shoots
and roots, free proline content increased, suggesting that treatments (2) and (3)
were capable to protect plants by decreasing oxidative stress. In shoots, treatments
(2) and (3) lead to a significant increase of GR activity (1.3x and 2.6x, respectively).
Contrastingly, GSH accumulation decreased significantly by 1.1x and 1.3x in treatments
(2) and (3), respectively. In roots, treatment (2) led to a significant decrease in both
GR activity and GSH levels (1.1x and 1.2x for GR and GSH, respectively), and so did
treatment (3) (2x and 1.6x for GR and GSH, respectively). These changes were always
131
more pronounced in treatment (3) in both organs analyzed. Knowing that GSH is a
Bioremediation
key element for cell homeostasis, its decrease may be associated with an increased
glutathione-S-transferase activity that may be eliminating the excess of PRG. GSH
Effect of aluminum on ros content and antioxidant system in rye (Secale
may be also decreasing the amount of ROS produced, since there was a decrease in
cereale l.) leaves and roots
H2O2 accumulation in treatments (2) and (3). Taken together, these results strongly
Alexandra de Sousa1; Manuela Matos2; Hamada AbdElggawad3; Han Asard3; Jorge Teixeira1;
Fernanda Fidalgo1
indicate that progesterone did not induce stress to the plants. Moreover, all results
suggest that the use of EBL together with PRG synergistically decrease oxidative
damage, which may translate in a higher phytoremediation capacity.
Aluminum rhizotoxicity is a major problem worldwide. However, the cause of Al
toxicity remains unclear as well as the complex metabolic processes inherent to its
tolerance in several plant species, such as rye (Secale cereale), which is considered the
1BioISI - Biosystems & Integrative Sciences Institute, Departamento de Biologia, Faculdade de Ciências,
Universidade do Porto, Rua Campo Alegre s/n, 4169-007 Porto, Portugal
most Al-tolerant cereal among Triticeae [1, 2]. In this study, we analyzed the Al effects
on plant biomass, ROS production (O.-2 and H2O2) and antioxidant defense system
(SOD, CAT and proline) using two rye genotypes, previously classified as tolerant and
sensitive to this metal [3]. Seeds of both genotypes were germinated in the dark
at 25ºC and then transferred to a modified Hoagland´s solution and maintained
under a photoperiod of 16h of light at 25ºC. The nutrient solution was continuously
aerated and the pH was maintained at 4.0. Seedlings’ treatment with Al and samples
for analysis were collected as previously described [3]. In order to evaluate Al effects
in the seedlings, plant biomass production was determined as fresh weight. In
leaves and roots a higher increase in fresh weight throughout the Al treatment in
the tolerant genotype was observed. Al exposure resulted in the increase of O.-2 and
H2O2 contents in leaves and roots of the sensitive genotype when compared to the
tolerant one. O.-2 levels were higher in roots than in leaves in the sensitive genotype.
An opposite situation was observed for H2O2 content since this ROS was accumulated
primarily in leaves of both genotypes. Activity assays of SOD and CAT showed that
these enzymes increased in both genotypes throughout the Al treatment. Enzyme
activities were more evident in the tolerant genotype in both leaves and roots. It was
133
also observed that proline content was higher in leaves from both genotypes. These
Bioremediation
data point to a role of these enzymes as well as of the antioxidant proline in rye Al
tolerance under short time exposure to this metal. Evaluation of other components
Bioremediation of different types of oil in estuarine and coastal environ-
of the rye antioxidant system including antioxidant metabolites against Al-induced
ments – the role of autochthonous microorganisms
oxidative stress in both genotypes are in progress.
Vanessa Gouveia1; C. Marisa R. Almeida2; Ana P. Mucha2
[1] Ryan, P.R., Tyerman, S.D., Sasaki, T., Furuichi, T., Yamamoto, Y., Zhang, W.H., Delhaize, E. (2011). The
identification of aluminium-resistance genes provides opportunities for enhancing crop production on
acid soils J Exp Bot. 62: 9-20.
[2] Aniol, A., Gustafson, J. P. (1984). Chromosome location of genes controlling aluminum tolerance in
wheat, rye, and triticale. Can J Genet Cytol. 26: 701-705.
[3] A. de Sousa, M. Matos, J. Teixeira, F. Fidalgo. 2014. ´´Membrane oxidative damage in roots and leaves
of rye seedlings exposed to aluminum´´. VI Jornadas Nacionais de Genética e Biotecnologia e II Encontro
Nacional de Estudantes de Biotecnologia. Universidade de Trás-os-Montes e Alto Douro, Vila Real, Portugal
1Biosystems & Integrative Sciences Institute (BioISI), Departamento de Biologia, Faculdade de Ciências,
Universidade do Porto, Portugal;
2Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD), Vila
Real, Portugal;
3Laboratory for Molecular Plant Physiology and Biotechnology, Department of Biology, University of
Antwerp, B-2020 Antwerp, Belgium
Bioremediation can be a valuable tool to recover ecosystems affected by oil spills.
However, the severity of oil spills impact depends on the type of oil, the size of the
spill and the ecosystem affected factors that can also influence bioremediation
performance. The present work aim to study the potential role of autochthonous
microorganisms for bioremediation of sediments contaminated with different types
of oil. Sediments were collected in 3 locations along the northern Portuguese coast:
in a sandy coastal beach and in Minho and Douro estuaries. Laboratory microcosms
(50 ml flasks) experiments were carried out with sediments spiked with crude oil,
diesel oil or turbine oil. Microcosms were incubated in Bushnell Haas medium
supplemented with sodium chloride and nitrogen, under constant shake for 15 days.
At the beginning and at the end of the experiment, sediments were characterized
in terms of microbial community structure (by ARISA), hydrocarbon degrading
microorganisms’ abundance (HD by MPN) and Total Petroleum Hydrocarbons
concentrations (by FTIR). At the beginning of the experiment, significant differences
among HD were observed, with lower abundances in the sandy coastal beach and
higher abundances in the Douro estuary. Nevertheless, after 15 days of exposure to
the different types of oil, all microbial communities increased their HD and, in general,
no significant differences were observed among sediments. Results obtained so far
indicate that the studied autochthonous microbial communities were able to adapt
to the different types of oil by increasing their abundance in hydrocarbon degraders.
135
Analyses are still in course to assess changes in the microbial community structure
Emerging Toxic Compounds
and their capacity to degrade hydrocarbons.
Caracterización de la fotodegradación de fármacos en sistemas acuátiAcknowledgments
To the Project ECORISK (reference NORTE-07-0124-FEDER-000054), co-financed by the North Portugal
Regional Operational Programme (ON.2 – O Novo Norte), under the National Strategic Reference
Framework (NSRF), through the European Regional Development Fund (ERDF).
1CIMAR/CIIMAR – Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto;
Instituto de Ciências Biomédicas Abel Salazar Universidade do Porto;
2CIMAR/CIIMAR – Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto
cos mediante cromatografía líquida-espectrometría de masas
R.M. Baena-Nogueras1; E. González-Mazo1; P.A. Lara-Martín1
Hoy en día se usa una gran cantidad de productos farmacéuticos tanto en pacientes
humanos como en veterinaria, los cuales pueden alcanzar diversos sistemas acuáticos
tras su utilización y posterior vertido. Así, podemos detectar concentraciones (en
el orden de ppt a ppb) de estos micro-contaminantes no sólo en aguas residuales
sino también en aguas superficiales y, ocasionalmente, en agua potable. Uno de
los principales factores responsables de la eliminación de estos compuestos en el
medio ambiente es la acción de la luz solar natural, la cual puede contribuir a la
degradación de fármacos por mecanismos directos (absorción de la energía solar
directamente en la molécula) y/o indirectos (interacción de reactivos intermedios
generados como los radicales hidroxilos). Este trabajo se centra en caracterizar la
cinética de fotodegradación de un amplio grupo de fármacos (> 80, incluyendo 50
antibióticos) en aguas naturales en un rango de pH relevante medioambientalmente
(de 4 a 9). El seguimiento de la cinética de degradación se ha llevado a cabo mediante
cromatografía de líquidos de ultra-resolución acoplada a espectrometría de masas
en tándem (UPLC-MS-MS). Se han obtenido las constantes de fotodegradación (k)
tras ajustar los datos a una cinética de primer orden, encontrándose un amplio
rango (entre 0.0001 min-1, en el caso de gemfibrozil, y 0.5 min-1 para ketoprofen)
así como los tiempos de vida media (t1/2) de todas las sustancias investigadas
(oscilan entre 1 min y > 100 h según el compuesto). Gran parte de los fármacos,
especialmente antiinflamatorios y antibióticos, se degradaron considerablemente
137
(ej.: >50% en 3.5 horas) a excepción de los de tipo macrólido y penicilinas. Además,
Emerging Toxic Compounds
se observó un aumento generalizado en la constante de fotodegradación a valores
de pH inferiores a 7, si bien algunos compuestos presentaron mayor velocidad
Impact of regulated and emerging pollutants and microplastics in
de degradación a pH básico (ej.: penicilinas, lincosamidas). Posteriormente se
marine ecosystems (IMPACTA project)
seleccionó aquellos compuestos que presentaron las tasas de fotodegradación más
Víctor M. León1; Soledad Muniategui2; Ricardo Beiras3; Juan Bellas4; Jesús Gago4; Juan A. Campillo1;
Concepción Martínez-Gómez1; Lucía Viñas4; Victoria Besada4; José Fumega4; J.M. Andrade2; J.
Moreda2; E. Beceiro2; R. Soto2; E. Alonso2; M.J. González2; Paula Sánchez-Marín3; Leticia Vidal3
elevadas para identificar los fotoproductos resultantes mediante espectrometría de
masas de alta resolución (UPLC-q-ToF-MS), proponiéndose por primera vez la ruta
de fotodegradación para multitud de compuestos (36) pertenecientes al grupo de
antiinflamatorios (ej.: ketoprofeno, naproxeno, ibuprofeno etc.) y antibióticos de
diverso tipo (ej.: quinolonas, sulfonamidas, cefalosporinas, nitroimidazoles).
Marine ecosystems are nowadays subjected to a massive input of synthetic
chemicals from everywhere. Unfortunately only a small portion of them is being
monitored, and it is necessary to identify which pollutants can produce adverse
impacts in the marine environment. IMPACTA project (CTM2013-48194-C3) is
1Universidad de Cádiz. Departamento de Química Física, Facultad de Ciencias del Mar y Ambientales,
characterizing the distribution of regulated and emerging contaminants and
Universidad de Cádiz, Campus Río San Pedro s/n, Puerto Real (Cádiz), 11510, España.
microplastics in marine sediments, and evaluating the biological effects that
they can cause (sing sublethal embryotoxicity tests, endocrine disruption and
biomarkers). Sensitive and selective analytical methods are being developed and
validated for pharmaceuticals, perfluorinated compounds, organophosphorus
pesticides, triazines, personal care products, nonylphenols and alkylated PAHs
in sediments. Thus, relevant pollutants present in coastal and offshore areas will
be identified. Furthermore potential toxic effects of the contaminants present
in coastal sediments are being assessed through embryotoxicity bioassays and
also the biological effects on different marine species as a consequence of their
exposition to specific compounds.
Another relevant contribution of this project will be the assessment of
the impact of micro-plastics, first time in the Spanish coastal areas. Their
potential toxic effects and their role in the transference of pollutants in the
marine environment are being assessed. The concentration and composition of
139
microplastics in sediments and demersal fish stomachs are being characterized,
Emerging Toxic Compounds
and their potential biological effects on marine invertebrates are also being
investigated.
Concerning pharmaceuticals, what are the portuguese wastewater
treatment plants hotspots of contamination?
Acknowledgements
This work was supported by the Spanish Inter-Ministerial Science and Technology Commission through
the ‘IMPACTA’ (CICYT, CTM2013-48194-C3-1-R) project and the European Union through the European
Regional Development Fund (ERDF).
1Instituto Español de Oceanografía, Centro Oceanográfico de Murcia;
2Departamento de Química Analítica, Facultade de Ciencias, Universidade da Coruña;
3Departamento de Ecoloxía e Bioloxía Animal, Universidade de Vigo
4Instituto Español de Oceanografía, Centro Oceanográfico de Vigo
A.M.P.T. Pereira1; L.J.G. Silva1; C.M. Lino1; L.M. Meisel2; A. Pena1
The presence of pharmaceuticals in the environment is a growing problem that
must be tackled to meet the Water Framework Directive of the European Union. As its
use cannot be avoided, a sound risk assessment of their presence in the environment
is a key issue (Ghiani et al., 2014; Robles-Molina et al., 2014).
In line with the Directive 2013/39/EU a Portuguese nationwide monitoring based
exercise, for the selection of the most representative surface waters was performed. To
meet this purpose, since the major point sources of pharmaceuticals environmental
contamination are wastewater treatment plants (WWTPs), the occurrence and fate
of eleven of the most consumed pharmaceuticals, belonging to several therapeutic
classes, in 15 WWTPs effluents (WWEs) from five different regions during one year (4
sampling campaigns) was assessed.
The analytical methodology used was based on solid phase extraction through
Oasis MAX and liquid chromatography with tandem mass spectrometry and method
detection limits ranged from 0.4 to 60.0 ng L-¹.
The evaluation of the most representative WWTPs was performed using the
amount of pharmaceuticals released by each WWE and river flow.
All samples were contaminated with at least one and up to 8, from the 11 targeted
pharmaceuticals. The highest contaminations observed were 33 µg L-1, 1.649 kg day-1
and 64.2 ng L-1 concerning WWE concentration, the amount released per day by each
WWTP and the predicted surface water contamination, respectively.
141
Observing the predicted surface water contamination and eliminating the
Emerging Toxic Compounds
discharges in the Atlantic Ocean, this study highlights that the rivers Mondego,
Tagus, Ave, Trancão, Fervença and Xarrama should be selected as surface water
Removal of veterinary antibiotics in constructed wetlands microcosms –
monitoring stations.
response of microbial community
This study gives a good overview on pharmaceuticals contamination in WWTPs
Filipa Santos1; C. Marisa R. Almeida2; I. Ribeiro1; A. Catarina Ferreira3; Ana P. Mucha2
and its impact on surface waters in Portugal, supporting the selection of the 6 most
representative monitoring stations in Portugal.
The environmental fate of pharmaceuticals has been receiving increasing
attention, especially since the detection of their presence in wastewater treatment
Acknowledgments
The authors thank FCT the financial support (project and fellowship PTDC/AAC-AMB/120889/2010,
fellowship granted to L.J.G. Silva SFRH/BPD/62877/2009); Instituto da Água da Região do Norte (IAREN) of
Portugal, for the MS analyses; every entity that provided technical assistance in collecting the wastewater
samples.
plant (WWTP) effluents, surface and groundwater. These compounds can be released
into nature directly, by discharge or inadequate treatment of water, or indirectly,
when manure is used as agriculture fertiliser. Within the veterinary drugs, antibiotics
are the group of most concern due to the risk of spread of antibiotic resistance in
the environment. In addition to their use to treat diseases, antibiotics are extensively
References
Ghiani, M., Tavazzi, S., Mariani, G., Locoro, G., Loos, R., Parachini, B., Sena, F., Surkuusk, G., Gans, O., Wulf, E.
De, Feren, M., Ternes, T., Wick, A., Belli, K.M., Stroomberg, G., Rand, R., Thomas, J., Thomas, R., Walmsley, R.,
Whalley, C., Gawlik, B.M., 2014. Feasibility of a Monitoring Mechanism Supporting a Watch List under the
Water Framework Directive.
Robles-Molina, J., Lara-Ortega, F.J., Gilbert-López, B., García-Reyes, J.F., Molina-Díaz, A., 2014. Multi-residue
method for the determination of over 400 priority and emerging pollutants in water and wastewater by
solid-phase extraction and liquid chromatography-time-of-flight mass spectrometry. J. Chromatogr. A
1350, 30–43.
1LAQV, REQUIMTE, Group of Bromatology, Pharmacognosy and Analytical Sciences, Faculty of Pharmacy,
University of Coimbra, Polo III, Azinhaga de Stª Comba, 3000-548 Coimbra, Portugal;
2INFARMED, I.P. - National Authority of Medicines and Health Products, 1749-004 Lisboa, Portugal
used as enhancer of feed efficiency and as growth promoters. Constructed wetlands
(CWs) are artificial complexes that can be used as alternative or additive low-cost
wastewater treatments, for the removal of several contaminants. Considering that
microorganisms can have a key role in CWs and that the microbial communities can
be affected by a diversity of contaminants, the present study aimed to evaluate the
response of microorganisms from CWs microcosms to the presence of veterinary
antibiotics, both in terms of community structure and removal performance. Three
sets of microcosms planted with Phragmites australis (3 replicates each) were run
in parallel: a set was feed only with livestock wastewater and two sets were feed
with the same wastewater doped with antibiotics (enrofloxacin or ceftiofur at 100
µg/L). Wastewater was treated during 18 one-week cycles. After each one-week
cycle wastewater was removed and replaced by new one (doped or not). Water and
sediment samples were collected at the end of week 1, 2, 4, 8, 14 and 18. Antibiotics
143
removal was evaluated by HPLC while microbial community was characterized
Microbial Ecotoxicology
by ARISA and by 454-pyrosequencing. Results show that microbial communities
were dominated by the phylumProteobacteria (38 to 48%), Firmicutes (20 to 27%)
Relative sensitivities of life history variables of freshwater zooplankton
and Bacteroidetes (12 to 15%), independently of the presence of the antibiotics.
(rotifers and cladocerans) to stress: a review
The study also shows that the systems were able to remove more than 90% of the
S.S.S. Sarma1; S. Nandini2
added antibiotics, pointing to the applicability of CWs for the removal of veterinary
antibiotics from livestock wastewaters.
Freshwater zooplankton species including rotifers and cladocerans live under
constant stress both natural (temperature, salinity, food limitation, proliferation of toxic
Acknowledgements
To European Regional Development Fund through COMPETE - Operational Competitiveness Program and
national funds through FCT, under PEst-C/MAR/LA0015/2013.
1CIMAR/CIIMAR,
Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto;
Instituto de Ciências Biomédicas Abel Salazar Universidade do Porto;
2CIMAR/CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto;
3CIMAR/CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto;
Faculdade de Ciências, Universidade do Porto
cyanobacteria etc.) and man-made (pesticides, heavy metals, pharmaceuticals) factors.
When exposed to toxicant stress, both survivorship and reproduction-related variables of
zooplankton are affected. However, under sublethal levels of toxicants, the reproductive
variables appear to be more sensitive than survival-related variables. When subjected to
temperature stress, the lifespan of zooplankton enhances at lower levels or decreases if
the intensity is higher. Neonates are generally more sensitive than adults. For freshwater
zooplankton, salinity is one of the strongest natural stresses, which normally affects the
survival than reproduction. The effect of herbicides on zooplankton is different from that
of pesticides. Similarly, zooplankton species appear to tolerate higher concentrations of
certain heavy metals (e.g., Zn at mg/L levels) than others (e.g., Hg in µg/L levels). Here we
compared the response of life history variables (lifespan, generation time, gross and net
reproductive rates and rate of population increase) of zooplankton species to selected
natural and man-made stresses. Comments have been made for different methods used
in literature (partial life table, multi-generational approach, differences among species
and clones etc.) for quantifying the response of zooplankton to stresses.
1Universidad Nacional Autónoma de México;
2Laboratory of Aquatic Zoology, Universidad Nacional Autónoma de México
145
Microbial Ecotoxicology
Effect of mixed toxic diets (Microcystis aeruginosa and Scenedesmus acutus)
on the competition of two cladocerans (Daphnia cf. mendotae and Simocephal)
Alfredo Pérez Morales1; C.A. Espinosa-Rodríguez2 ; L. Rivera-De la Parra2; S.S.S. Sarma2; S. Nandini2
Microcystis aeruginosa is a common bloom forming cyanobacteria in eutrophic
lakes of Central Mexico. In nature, zooplankton groups including cladocerans
suffer from natural toxic substances produced by the cyanobacteria, especially M.
aeruginosa. In this regard, several works kave focused towards effects on somatic
growth, survival and fecundity of different cladoceran species; however, information
about competition among cladocerans under cyanobacterial blooms is less known.
Green algae such as Scenedesmus inhabit the same waterbodies where Microcystis
occurs as blooms. So, it is then possible that cladocerans feed on mixed diets
containing algae and cyanobacteria. In this work competitive outcome between two
cladocerans (Daphnia cf. mendotae and Simocephalus mixtus) were quantified using
mixed diets. Cladocerans were fed sonicated Microcystis aeruginosa alone (100%) or
mixed with Scenedesmus acutus (0, 25, 50 or 75%, based on cell density); the total
cell density (cyanobacteria, green algae or their mixed form) offered daily per test jar
was 0.5x106 cells/mL. In controls where each zooplankton species was grown alone,
the population growth of cladocerans increased with increasing proportion of green
algae (100 or 75%) in diet. In mixed cladoceran cultures D. mendotae was dominant
on S. mixtus in all mixed diets tested. Our results showed that S. mixtus was more
susceptible to the presence of M. aeruginosa regardless the proportion of S. acutus in
the diet. This study indicates that mixed diets of M. aeruginosa and S. acutus affect the
147
competitive outcome in cladocerans. Thus, there were species-specific differences in
Microbial Ecotoxicology
population growth when offered mixed diets of green algae and toxic cyanobacteria.
These probably explain the coexistence of different cladoceran species in Microcystis-
Solid lipid nanoparticles affect leaf litter microbial colonizers and enzy-
dominated waterbodies in Central Mexico.
matic activities in freshwater
Ana Sampaio1; R.J. Mendes2; P.G. Castro2; S. Doktorovova2; A.M. Silva1
1Instituto de Ciencias Marinas y Pesquerías;
2Universidad Nacional Autónoma de México, Campus Iztacala, Av. De Los Barrios s/n, Los Reyes Iztacala,
Tlalnepantla, Estado de México. C.P. 54090. México
Solid lipid nanoparticles (SLNs) are used as carriers for drug delivery, and are high
biocompatible and designed to endure in the host organism. Despite its current
industrial production is low, many of these substances are available on the market,
and much more are in the production pipeline. As a result, many of them will end in
aquatic systems, sometimes metabolically unchanged, raising the question whether
they can pose a risk to aquatic biota and the associated ecological processes. Microbial
decomposers of plant litter, play a key role in forested streams being responsible for
the energy flow between terrestrial and aquatic environments. Here, we investigated
the effects of SLNs on alder leaf litter decomposition by aquatic microbes. Alder leaves
were immersed in a stream of Northeast Portugal to allow microbial colonization
before being exposed in microcosms of two types of SLNs at two concentrations for
42 days. Results showed that rates of leaf decomposition decreased with exposure
to SLNs. Bacterial biomass was not inhibited by SLNs, contrary to cultivable fungi.
The type and concentration of SLNs influenced differently the leaf colonization
by yeast and filamentous fungi as well the sporulation rate of fungi. These effects
were accompanied by changes in the community enzymatic profile: the activities
of alkaline phosphatase, acidic phosphatase, Napthol-AS-BI-phosphohydrolase and
lipases increased in the SLNs microcosms. Overall, results indicate that the release
of SLNs to the environment may affect microbial communities with impacts on leaf
litter decomposition in streams, and that those effects depend on SLNs composition
149
and concentration, as well on the microbial target group. Thus prior to massive
Microbial Ecotoxicology
industrial production of these nanomaterials some measures should be taken to
avoid environmental impact affecting these microbial communities.
Effect of caffeine on the life table demography of the rotifer Plationus
patulus and the cladoceran Moina macrocopa
1Department of Biology and Environment, University of Trás-os Montes e Alto Douro (UTAD) | Centre for
Research and Technology of Agro-Environmental and Biological Sciences;
2Department of Biology and Environment, University of Trás-os Montes e Alto Douro (UTAD)
S. Nandini1; M. C. Reyes-Santillán2; S.S.S. Sarma2
Eutrophication is common in several water bodies in Mexico, mainly because
they receive partially or untreated waste water. Caffeine is an indicator of
anthropogenic wastes which tends to persist in water. Although it may have
a negative effect on aquatic organisms, little information is available on this
subject. Zooplankton species are sensitive indicators of water quality. In this
study we tested the effect of sub-lethal concentrations of caffeine to the rotifer
Plationus patulus and one of the most common cladocerans in tropical systems,
Moina macrocopa. To derive the median lethal concentration, clonal populations
of both species were used. The zooplankton species were maintained on a diet
of Chlorella vulgaris cultured on defined medium and moderately hard water.
Using stock solution of caffeine (1g L-1), different concentrations of caffeine (25,
50, 100, 150, 200 and 250 mg L-1) were prepared and distributed separately into
10 ml jars containing 10 females (neonates) for each zooplankton species. Each
treatment had four replicates; controls without caffeine were also set up. Mortality
was estimated after 24 h and the LC50 was derived using Probit method. Moina
macrocopa and Plationus patulus were resistant to caffeine; the LC50 for M.
macrocopa was 141 mg L-1 and for Plationus patulus no mortality was observed
within 24 hours at concentrations up to 500 mg L-1. However, chronic exposure
to caffeine resulted in a decrease in the demographic variables of both species.
Growth rates ranged from 0.3 to 0.35 d-1 for the rotifer and 0.2 to 0.42 d-1 for the
151
cladoceran. Although concentrations of caffeine in natural environments are low,
Ecosystem Level Effects
a constant long-term exposure may affect zooplankton populations.
A new tool for water quality assessment combining distinct levels of orUniversidad Nacional Autónoma de México;
2
Laboratory of Aquatic Zoology, Universidad Nacional Autónoma de México, Campus Iztacala, Av. de los
Barrios No.1, AP 314, Código Postal 54090, Los Reyes, Tlalnepantla, Estado de México, México
1
ganization
Rui Cortes1; Samantha Hughes1; Ana Coimbra1; João Carrola1; Sandra Mariza Monteiro1; Ana Pinto1;
Sandra Pereira1; Ana Sampaio1; Joaquim de Jesus1; Vitor Pereira1; Catia Santos1; Marisa Lopes1;
Simone Varandas1
Distinct leveles of organization were combined to improve water quality
assessment under the Water Framework Directive, namely the ones expressing
global ecological information based on bio-indicators or on more specific causeeffect relations derived from biomarkers. This new and integrative approach took
place in different streams of North Portugal, covering a wide range of conditions,
from relative pristine aquatic ecosystems to highly impacted ones, allowing the
the observations along a clear gradient of disturbance. Two communities were
used _fishes and invertebrates_but also it were included descriptors of river
functioning. In those communities we took in consideration a whole set of biological
parameters involving different biological levels of organization, from lower scales
related to biomarkers of exposure and effec, like histopathological and biochemical
modifications in specific organs (liver and fish gills) to higher scale analysis, based on
bio-indicators, such as metris and traits of invertebrates (including Chironomidade
exuviae) and fish guilds. At the highest level it were used parameters associated to leaf
litter breakdown, with a significant importance on energy transference in the system.
Such a high number of variables were treated by Forest Random Classification, taking
into account the different groups with distinct ecological state. Therefore, a total of
59 variables emerged as the ones that were more consistent in discriminating those
ecological states, allowing also to determinate for each of those variables the specific
153
cutting levels separating the different ecological class. A Multiple Ecological Level
Ecosystem Level Effects
Index (MELi) was then built consisting on a powerful tool to assess the ecological
condition in running waters since it integrates multiples sources of disturbance;
Inter-annual variability of soft bottom macrobenthic community of the
besides it expresses the deviation from the reference condition which is according
NW Gulf of Mexico in relationship with the DWH oil spill
to WFD. Moreover, the index is flexible according to the information available and
Diana Salcedo1; Luis A. Soto1; A. Estradas1; A. V. Botello1
consequently each set of data may be analysed separately.
Benthic organisms have long been recognized as excellent bio-indicators of
UTAD
1
adverse conditions in marine ecosystems. Variations in their community structure
can reveal changes in environmental conditions related to natural or anthropogenic
disturbances. On April 2010, a massive oil spill occurred in the northern Gulf of
Mexico (GoM), caused by the sinking of the Deepwater Horizon (DWH). As a response
to this, a 3-year research program was undertaken in 2010, to assess the footprint of
the DWH blowout and its possible disturbance upon the soft-bottom macrobenthic
community inhabiting the shelf and upper continental slope of the NW GoM within
Mexico´s Exclusive Economic Zone. Community properties (abundance, density,
and biomass) of the benthic component and its temporal and spatial variability
were analyzed applying the PERMANOVA routine; particular attention was given
to stressing toxicant parameters such as polycyclic aromatic (PAHs), aliphatic
hydrocarbons (AHs) and heavy metals in the studied area. Overall density values
tended to increase (163 – 183 - 451 ind/10cm2) though the proportion of taxa
changed, and small-size opportunistic organisms (polychaetes and nematodes)
became dominant throughout the study. Estimated annual abundance-biomass
curves (ABC) revealed progressive stress scenarios from moderate to severe
towards2012. The concentration of geochemical variables inherent to oil (vanadium,
nickel, cobalt, PAHs and AHs) showed an increasing trend over time. However,
the BIO-ENV routine exhibited low correlations between density and preselected
155
biogeochemical variables, out of the 23 determined in this study. Initially, edaphic
Ecosystem Level Effects
properties seemed to regulate the benthic community stability; afterwards, stressful
variables, indicative of a possible anthropogenic effect upon the community (nickel,
Free-living nematodes as bio-indicators of the DWH oil spill in the mexi-
vanadium, and PAHs), were highlighted by BIO-ENV. The interannual variability
can exclusive economic zone, NW Gulf of Mexico
observed in macroinfauna of the NW Gulf is the result of the synergy of several
Luis A. Soto1; Diana Salcedo1; Karina Arvizu1; Alfonso V. Botello1
environmental factors. Undoubtedly, compounds derived from fossil fuels have a
significant disturbance role. Their possible source, either local or regional remains
debatable.
The presence and abundance of certain nematode species in marine sediments
are good bio-indicators of environmental disturbance, and it can even reveal the
existence of specific toxicants. We studied this important infaunal component in
1 Instituto de Ciencias del Mar y Limnología, UNAM, México
D.F. 04510 MÉXICO.
a 3-year monitoring program in Mexico’s EEZ in the aftermath of a major oil spill
in the northern Gulf caused by the accidental sinking of the Deepwater Horizon
platform (DHW) in April of 2010.Some nematodes are known for their tolerance to
hydrocarbon compounds derived from fossil fuels. A closer look at the nematode
composition and density values throughout the DWH oil spill revealed considerable
changes. Continental shelf and upper slope sediments sampled in the summer of
2010 included 48 genera and a density of 100 ind/10 cm2 that represented 62 %
of the total macroinfauna. Eight months later in the winter of 2011, there was a
critical decrement in genera number (23) and density (60 ind/10 cm2) equivalent to
33 % of the total macroinfauna. However, in 2012 this community showed signs of
a recovery. Genus diversity was 58 with a density of 156 ind/10 cm2 or 35 % of the
total macroinfauna. Despite the unequal number of sampling sites covered during
the three observational periods, no significant differences (p> 0.05) were detected
in nematodes depth or longitudinal distributional patterns. Interestingly, early in
this study, both the macroinfauna and the nematode community displayed a similar
response to the influence of individual geochemical variables namely, aluminum, total
nitrogen and polycyclic aromatic hydrocarbons (PAHs) concentrations. Afterwards,
157
nematodes abundance and composition in 2011 were altered due to the presence
Environmental Chemistry Monitoring
of PAHs and magnesium in sediments. In contrast, in 2012, small concentrations of
Al and the prevalence of PAHs seem to promote the dominance of nematodes such
Microplastics in coastal sediments from South Portugal
as Sabatieria sp, known for its tolerance to high levels of hydrocarbons and heavy
Jesus Gago1; J.P.G.L.Frias2; P.Sobral2; V.Otero3
metals.
Coastal sediment samples were collected by divers of the Portuguese Task Group for the
1 Instituto de Ciencias del Mar y Limnología, UNAM, México
D.F. 04510 MÉXICO.
Extension of the Continental Shelf (EMEPC), during the 2013 M@rbis campaign, between
June and August, in Algarve coastal waters. Microplastics were retrieved from processed
sediment samples, in order to estimate amounts and polymer type identification.
A total of 31 microplastics (25 fibres and 6 fragments) were collected from the
coastal sediment samples. Fibres of 4 different colours (red, green, blue and black)
and fragments of two colours (blue and green) were found. The overall percentage of
microplastics per volume of sediment is 0.24%, and the number of microplastics per
weight of dry sediment is 0.01 microplastics g-1. The vast majority of the samples were
identified as a semisynthetic cellulose based polymer, commonly known as Rayon.
Rayon is the oldest commercial manmade fibre composed of regenerated cellulose.
Rayon fibres are engineered to possess a range of properties, from flame retardants to
super absorbent fibres, to meet the demands for a wide variety of end uses.
These results show for the first time, to the best of our knowledge, the presence
of microplastics in coastal sediments in the South of Portugal. This study provides
an overview of microplastic pollution in the Southern Portugal coast by reporting
microplastic concentration and polymer type, in the surveyed area.
1Instituto Español de Oceanografía;
2MARE- FCT-UNL;
3REQUIMTE-LAQV- FCT-UNL
159
Environmental Chemistry Monitoring
Polychlorinated biphenyls in molluscs, within basque estuaries (northern
Spain): relationships with hydrological characteristics and human pressures
Oihana Solaun1; J. German Rodriguez1; Angel Borja1; Joana Larreta1; Victoriano Valencia1
Interannual variability of polychlorinated biphenyls (PCBs) (IUPAC numbers
28, 52, 101, 118, 138, 153 and 180), measured in the soft tissues of Mytilus
galloprovincialis mussels and Crassostrea gigas oysters, collected from estuarine
waters within the Basque Country (Bay of Biscay), is investigated. Samples
were collected annually, in autumn, between 2002 and 2011. The results
have been analysed statistically, applying the Mann-Whitney-Wilcoxon test,
for comparison between the median concentrations in mussels and oysters.
Moreover, ReDundancy Analysis (RDA) was used to evaluate the relationship
between the median concentrations of the PCB congeners per site, during the
period considered, with some independent variables (including hydrological
characteristics and human pressure of the estuaries wherein they are
located). The results show significant (p<0.05) differences between median
concentrations in mussels and oyster, for some of the congeners (i.e., CBs 101,
138, 153 and 180) and for Sum7PCB. Sites located within the ports of Bilbao
and Pasaia showed the highest PCBs concentrations in molluscs; the lowest
were observed in the mouth of the Oka estuary, an area of low population and
industrial activity. Congener profiles of PCBs in the tissues of molluscs reveal
the predominance of hexachlorobiphenyls (CB153 and CB138). In addition,
redundancy analysis has shown that residence time, river flow and a “pressure
index” explain 57% of the variability in the PCB congener concentrations (the
161
higher the values of these variables, the higher the concentration). Finally,
Environmental Chemistry Monitoring
Sum7PCB median concentrations in molluscs were correlated to median
concentrations in sediments collected from nearby sampling sites (r2=0.51,
Multibiomarker assessment of cadmium-based quantum dots effects in
p<0.01).
the marine mussel Mytilus galloprovincialis
Thiago Lopes Rocha1; Tânia Gomes2; Cátia Cardoso1; Nélia C. Mestre1; Vânia Serrão Sousa1; Maria
João Bebianno1
1AZTI
In recent years, quantum dots (QDs) have generated interest from the life
sciences community due to potential applications in nanomedicine, biology and
electronics. However, these engineered nanomaterials can be released into the
marine environment, where their ecotoxicological effects in marine organisms
remain unclear. The mussel Mytilus sp. is an important model to assess the toxicity
and environmental risk of nanomaterials in aquatic ecosystems. In this work, a
multibiomarker approach was performed to assess the ecotoxicological impact
of CdTeQDs and their dissolved counterpart, using the marine mussel Mytilus
galloprovincialis. Mussels were exposed in vivo to CdTeQDs (size: 6 ± 1 nm; 10 µgCd.L-1)
and dissolved Cd (Cd nitrate; 10 µgCd.L-1) for 14 days and a battery of biomarkers
was analyzed: immunotoxicity (density, viability and differential cell count of
hemocytes), cytotoxicity (LMS), genotoxicity (DNA damage and nuclear anomalies),
metal exposure (MT), oxidative stress (CAT, SOD, GST and GPx), oxidative damage
(LPO), neurotoxicity (AChE) and condition index along with the determination of
Cd concentration in mussel tissues. Results showed that Cd accumulation is tissuespecific and Cd form dependent, with dissolved Cd being the most bioavaliable
form. Both Cd forms modified biomarker response in mussels and different patterns
were obtained for nano and dissolved Cd. Mussels exposed to QDs showed lower
Cd accumulation, oxidative stress and LPO than those exposed to dissolved Cd.
QDs were classified as immunocytotoxic and genotoxic, but not cytogenotoxic. In
163
addition, QDs also changed biochemical defence systems of exposed mussels and
Nanotoxicology
induced MTs. Both Cd forms induced several effects at biochemical and cellular levels
highlighting their potential risk for marine organisms. The overall results indicate
Oxidative stress and enzymatic alterations induced by multi-walled carbon
that in vivo toxicity of both Cd forms are mediated by different modes of action and
nantoubes (MWCNTs) funcionalized with polyethylene glycol (PEG) in mouse
suggest that the mussel M. galloprovincialis is a suitable model for environmental risk
liver tissue
assessment of Cd-based QDs.
Silvia Pierre Irazusta1,2; Elaine Conceição de Oliveira1,3; Helder Ceragioli4; Bruno Fernando Santos
de Souza5; Monique Culturato Mendonça7; Edilene Siqueira Soares7; Romildo Aevedo Jr6; Maria
Alice Cruz-Hofling7; Zilma Maria Almeida Cruz8.
Keywords
Nanomaterials, oxidative stress, antioxidant system, biomarkers, molluscs.
As the field of nanotechnology develops, there are a crescente number of
products on the market with a claim to contain elements of nanotechnology. The
1University of Algarve;
2Norwegian Institute for Water Research (NIVA)
potential for public and occupational exposure is likely to increase, and so there is an
urgent necessity to consider the possibility of any detrimental health consequences
with this increased exposure to nanoparticles (NP). Studies have shown that (NP)
exert oxidative stress and cause severe lipid peroxidation and oxidative stress, which
has been implicated as a possible mechanism for nanoparticles toxicity. The present
investigation aimed to evaluate the toxicologic and histologic issues relating to
sistemic administration of two multi-walled carbono nanotubes (MWCNT) in liver. The
analyses realized were immunohistochemistry and western blotting for the SOD, CAT
and GST, RAMAN and FTIR and quantitative hystologic analysis. In our experiments,
NT1 produced more intense changes in hepatic enzymes in response to stress,
than NTL. There was CAT activation at the lowest dose, followed by depletion at the
highest dose, while there was activation of SOD. GST followed the same trajectory
of the CAT, which is consistent, since they are costimulated. SOD was overexpressed
by immunohistochemistry, with the expression of more than one isozyme. The most
significant changes were observed in the histologycal aspect, with alteration of
vascular diameter of centrilubulars veins, which showed significant enlargement of
the diameter by exposure to both CNTs, probably by a local cytotoxic effect. It was
165
also observed vacuolisation of hepatocytes around these veins.
Nanotoxicology
Conclusions: Our data support that CNT have a mild toxicity since that they only
produced disturbance in the endotelial environment, at the highest dose, proplably
Effects of multi-stressors on juveniles of the common goby (Poma-
via production of ROS.
toschistus microps): microplastics, nanoparticles and temperature
Pedro Ferreira1; Elsa Fonte1; Lúcia Guilhermino1
Keywords
Oxidadtive stress; MWCNT; PEG; liver; nanotoxicity.Abstract
1Faculty of Technology of Sorocaba - SP - Brazil;
2Postgraduate program in Management and Technology in Production Systems - State Center of Technical
Education in the State of Sao Paulo - Brazil;
3Laboratory of Neuroimmunology - Institutte of Biology - State University of Campinas - SP - Brazil;
4Faculty of Eletric Engineering and Computation - State University of Campinas - SP - Brazil;
5Federal University of São Carlos - SP - Brazil
6University of Vila Velha - ES - Brazil;
7Laboratory of Cellular Ultrastructure: Vennon Action in Biological Systems - Institutte of Biology - State
University of Campinas - SP - Brazil;
8Postgraduate program in Ecosystem and Ecology - ES – Brazil.
The contamination of the marine environment by nanomaterials and microplastics
(MP) has been raising an increasing concern in relation to both environmental and
human health. More knowledge on the effects of these environmental contaminants is
needed to improve the basis of ecological and human risk assessments, especially those
induced by simultaneous exposure to different substances (mixtures) and in relation to
alterations resulting from global climate changes (e.g. temperature increase). Thus, the
goal of the present study was to investigate the effects of gold nanoparticles (AuNPs),
alone and in mixture with MP, on juveniles of the common goby (Pomatoschistus microps)
at two distinct temperatures (20º C and 25ºC). Early juveniles of P. microps were collected
in the Minho River estuary and acclimated to lab conditions. The bioassays were carried
out at the two temperatures, by exposing juveniles individually to AuNPs, alone and in
mixture with MP, for 96h. After the exposure period, the predatory performance and sub
individual biomarkers were determined. The results indicated that exposure to AuNPs (low
ppb range) induces toxic effects on fish, and that temperature rise and the presence of
MP may interact with AuNPs. Therefore, more studies on the toxicity induced by mixtures
of emerging contaminants of high concern under temperature variation scenarios are
needed to improve the basis for environmental and human health risk assessments.
1ICBAS
& CIIMAR, University of Porto; ICBAS, Department of Populations Study, Laboratory of
Ecotoxicology,Rua de Jorge Viterbo Ferreira, 228,4050-123 Porto, Portugal; CIIMAR, Research Group of
Ecotoxicology, Stress ecology and Environmental Health
167
Nanotoxicology
Interaction of graphene nanomaterials with liver derived cell lines: internalization, cytotoxicity and potentiation of chemical effects
Tobias Lammel1; Paul Boisseaux1; María-Luisa Fernández-Cruz1; José M. Navas1
Graphene and its derivatives are increasingly produced and used in a wide
variety of products. In the case of agronomical applications of graphene, it can be
highlighted its potential utilization as a carrier of other substances to the inner of
cells. A consequence of the wide use of graphene is the increasing risks for humans
and wildlife. It is therefore essential to understand the mechanisms underlying the
possible toxic effects of graphene to cells and the processes guiding its cellular
internalization. Since liver plays a key role in detoxification processes, most
xenobiotics entering an organism will probably be conducted to this organ in an
attempt to metabolize them. The objective of this work was to study the toxicity of
two graphene derivatives (graphene oxide, GO, and carboxyl graphene, CXYG) to
two liver derived cell lines, HepG2 (human) and PLHC-1 (fish, Poeciliopsis llucida), and
to determine if graphene could serve as a carrier for other substances (polyaromatic
hydrocarbons, PAHs, were used as a model) to the inner of cells. GO and CXYG
suspensions in culture medium were thoroughly characterized. Cytotoxicity was
observed through three different assays informing about damage of plasma
membrane, mitochondria and lysosomes. Ethoxyresorufin-O-deethylase (EROD)
activity served to evidence cellular internalization of PAHs . Reactive Oxygen Species
(ROS) were also measured. Electron microscopy permitted observing the interaction
of graphene with cell membranes and the internalization process. GO and CXYG
caused a dose and time dependent cytotoxicity and were internalized in both cell
169
lines. In addition, GO and CXYG accumulated in the cells and were (secondarily)
Nanotoxicology
enclosed in vesicles. They caused an increase in ROS related with damages in the
mitochondrial membrane. Graphene derivatives also allowed the entrance of other
Mechanisms of action and toxicity of silver nanoparticles in model
contaminants into the cell by facilitating their diffusion and/or transporting them
aquatic and terrestrial organisms
across the plasma membrane. Acknowledgements: INIA project AT2011-001
Miren P. Cajaraville1; Nerea Duroudier2; Nerea García-Velasco2; José María Lacave2; Alvaro Fanjul2;
Mathilde Mikolaczyk3; Alba Jimeno-Romero2; Alberto Katsumiti2; Jörg Schäfer3; Eider Bilbao2;
Manu Soto2; Amaia Orbea2
1Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Departamento de Medio
Ambiente, Carretera de la Coruña, Km 7.5, 28040 Madrid.
In the last years, engineered nanoparticles (NPs) have been incorporated in many
consumer products, thus leading to growing concerns related to their potential
health and environmental hazards. Silver NPs are the most widely used metal NPs
due to their unique optical and catalytical properties and potent antimicrobial
activity. Further, Ag NPs resulted the most toxic metal NPs tested in previous studies
comparing a large set of different NPs. The present work aims to gain deeper
knowledge on the mechanisms of action and toxicity of Ag NPs (PVP/PEI-stabilized, 5
nm) in both aquatic and terrestrial animals. For this integrated approach, three model
organisms were employed: 1) mussels Mytilus galloprovincialis, filter-feeding bivalves
which are important targets of nanomaterials entering the water column and suitable
sentinels to monitor NP impact into the marine environment, 2) zebrafish Danio rerio,
a consolidated experimental model in biomedicine, at embryo and juvenile-adult
developmental stages, and 3) earthworms Eisenia fetida, terrestrial model organisms
widely used as bioindicators of soil pollution and as test organisms for standard
toxicity tests. Toxic effects of Ag NPs have been studied generally at concentrations
orders of magnitude higher than those expected in the environment, whereas in
this work experiments included environmentally relevant Ag NP concentrations.
Mussels and zebrafish were exposed through diet (algae Isochrysis galbana and
the crustacean Artemia sp, respectively), whereas earthworms were exposed to
171
artificially contaminated soils. ICP-MS and autometallography demonstrated Ag
Soil Pollution, Conservation and Restoration
bioavailability, mainly in digestive organs for the three species, in agreement with
Ag NP dietary exposure. Ag accumulation in mussels was higher in the restingperiod
Phytoavailability of lead in soils from an abandoned trap-shooting range
than in the reproductive period. Ag NP exposure elicited responses at cell and
Andrés Rodríguez-Seijo1; Daniel Arenas-Lago1; Manoel Lago-Vila1; María Luisa Andrade1; Flora
Alonso Vega1
tissue levels, including lysosomal membrane destabilization and genotoxicity, even
at environmentally relevant concentrations. Results obtained contribute to the
environmental risk assessment of Ag NPs.
There are over 100,000 shooting ranges in the world, which annually
deposited in soil million kilograms of Pb. This causes that shooting activities
are a problem of increasing importance in many countries because they
*Work supported by Spanish MINECO (MAT2012-39372), Basque Government (consolidated research
group IT810-13; Saiotek S-PE13UN142) and UPV/EHU (UFI11/37, PhD fellowships).
are the second source of Pb pollution. Moreover many of these facilities are
1University of the Basque Country;
2CBET Research Group, Dept. Zoology and Animal Cell Biology; Faculty of Science and Technology and
of this work were to analyze the contents of Pb in soils from a trap-shooting
Research Centre for Experimental Marine Biology and Biotechnology PIE, University of the Basque Country
UPV/EHU, Basque Country, Spain;
3Univ. Bordeaux, UMR 5805 EPOC, Allée Geoffroy St Hilaire, 33615 Pessac Cedex, France
located in the proximity of residential or agricultural areas. The objectives
range that becomes abandoned since 1999 and to study the effectiveness of
different extractants (CaCl2, DTPA, NH4OAc, LMWOA and BDW) for determining
its bioavailability. As spontaneous vegetation (Agrostis capillaris L.) is present
in the sampled area, the ability of A. capillaris as phytoremediator was also
evaluated. Thirteen soils from an old trap-shooting range (Galicia, NW Spain)
were sampled and analyzed. Soil main properties and total contents of Pb
were determined together with the operationally defined as available ones (by
means of the releasing solutions mentioned above). The results showed that Pb
contents in the soils are > 100 mg kg-1, all of them exceed the generic reference
levels for the soils of the region. In addition, in three of the soils, the Pb contents
are higher than the USEPA threshold level (400 mg kg-1). It was found that, in
general the reagent with greatest extraction efficiency was CaCl2, followed
by DTPA, NH 4OAc, LMWOA and BDW. The Pb contents in A. capillaris ranged
between 9.82 and 1,107.42 mg kg-1 (root) and between 6.43 and 135.23 mg
kg-1 (shoot). According to the metal content in the plants, CaCl2 is the releasing
173
solution that better represents the bioavailable content. The Pb accumulation
Environmental Chemistry and Toxicology
in roots and the presence of secondary mineral phases of metallic Pb in the
adjacent soil, showed the phyostabilizator properties of A. capillaris.
Assessment of trace elements pollution and its environmental risk in
freshwater sediments of the Alqueva Reservoir (Guadiana Basin)
Acknowledgements
This study was supported by the Ministerio de Economía y Competetividad (Project CGL2013-45494-R).
1Departamento de Biología Vegetal y Ciencia del Suelo. Lagoas. Marcosende. 36310, Vigo. Spain.
email: andresrodriguezseijo@uvigo.es
P. Palma1,2; L. Ledo1; P. Alvarenga1,3
The Guadiana Basin (SW Iberian Peninsula) is affected by acid mine drainage
(AMD), a consequence of ancient mining activities in the Iberian Pyrite Belt
(IPB). Consequently, the sediments at the Alqueva reservoir (SE Portugal) in
the Guadiana Basin are potentially contaminated by trace elements, which
makes important: (i) to characterize the status of trace elements pollution
of the sediments; (ii) to evaluate the mobility and the bioavailability of As,
Cd, Cu, Cr, Pb and Zn; and (iii) to assess the environmental risk associated
with the total and bioavailable concentrations of trace elements, using
the sediment quality guidelines (SQGs) and the risk assessment code
(RAC). Metal enrichment factors (EF) and geoaccumulation indexes (Igeo),
determined taking into account the regional background levels, revealed
that, among the metals analyzed, Cd contributed to the highest pollution
levels followed, by Pb and As. Despite the trace elements contamination of
the Alqueva sediments, the sequential extraction showed that most of them
are found in the oxidizable and residual fractions, which indicates that they
are sparingly bioavailable, with exception of Cd (acid-labile fraction) and Pb
(reducible fraction). Based on the RAC, Cd was the only metal that presented
a high risk, while Pb, As and Zn showed a medium risk. Moreover, the SQGs
revealed the existence of certain areas of extremely high risk, particularly
related to high concentrations of total As and, in less extent, of Pb and Cd,
175
associated with AMD, wastewater discharges and runoff of plant protection
Environmental Chemistry and Toxicology
products from agricultural fields located near the reservoir.
The coastal lagoons of Tabasco state, Mexico: endangered ecosystems
1Departamento de Tecnologias e Ciências Aplicadas, Escola Superior Agrária, Instituto Politécnico de Beja;
2CIMA - Centro de Investigação Marinha e Ambiental, CIMA, FCT, Edifício 7, Piso 1, Universidade do Algarve,
Campus Universitário de Gambelas, 8005-139 Faro;
3UIQA – Unidade de Investigação Química Ambiental, Instituto Superior de Agronomia, Universidade
Técnica de Lisboa.
Alfonso V. Botello1; Susana Villanueva1; Guadalupe Ponce1
Mexico posess 125 coastal lagoons on its litorals in the Gulf of Mexico, mexican
Caribbean and the Pacific Coasts. They represent important ecosystems for
aquaculture, fisheries, navigation and commerce, as well as energy exploitation
(petroleum) and urban and industrial settlements. In the south of the Gulf of Mexico,
are located the coastal lagoons of Tabasco state, (Carmen-Machona, Mecoacan and
Yucateco lagoons) where the mexican oil company (PEMEX) has been operating
since 40 years ago, producing petroleum pollution and singular impacts on fisheries
and mangrove ecosystems originated by the refuses of petroleum extracction and
refining.
Thus, studies conducting science 20 years ago, show a high petroleum pollution,
andpresence of toxic metals mainly chromium (Cr) in sediments and organisms
(fishes, crustaceans and mollusks) being of environmental risk for the populations
settle down in their nearbys. Trends in pollution seems to be improving, mainly due
to the better practices in exploitation of oil and legal regulation for refuses. Chemical
analysis for pollutans show that most of them are declining in concentrations, mainly
PAHs and toxic metals. Recently, another danger for these lagoons its represented
by the coastal erosion and vulnerability for meteorological events as the hurricans,
endangering not only the physical structures if not the human settlements.
1Institute for Marine Sciences and Limnology, National Autonomous University of Mexico (UNAM). Circuito
Exterior S/N , Ciudad Universitaria, México, D.F. CP 04510, MEXICO. gatoponcho@gmail.com
177
Environmental Chemistry and Toxicology
Is metal contamination in estuarine environments endangering european eel (Anguilla anguilla) in their first stage in continental waters?
Diana Castro1; Cláudia Mieiro1; João Pedro Coelho1; Sofia Guilherme2; Ana Marques2; Armando C.
Duarte1; Eduarda Pereira1; Mário Pacheco2
Estuaries are considered the most productive, as well as the most endangered
ecosystems, where anthropogenic activities contribute with significant loads
of mixtures of chemicals, directly affecting their natural functions. Several
diadromous migratory fish rely on estuaries’ high productivity, which offer
favorable conditions for their ontogenic migrations between marine and
riverine environments. This is the case of the European eel (Anguilla anguilla
L.), which is widely distributed across almost all the European territory. In
the last decades, the European eel population has demonstrated a steep
decline all over Europe, which has been attributed in part to pollution. This
work aimed at evaluating the genotoxic potential of mercury in A. anguilla
using the erythrocytic nuclear abnormalities (ENAs) assay. To provide
indirect information on the erythrocyte catabolism and erythropoiesis rate,
the frequency of immature erythrocytes (IE) and the nucleo-cytoplasmatic
ratio (NCR) were also assessed. This work was based on a field experiment
where glass eels were caged during 6 days along a recognized environmental
mercury gradient in the Ria de Aveiro, Portugal. Results showed that, despite
the frequency of ENAs was low, it was possible to distinguish an increasing
genotoxic risk at the contaminated sites. IE and NCR also indicated some
toxic potential at the contaminated sites. Hence, the results demonstrated
that mercury has the ability to induce genetic damage in glass eel blood cells
179
through clastogenic and/or aneugenic actions, suggesting a harmful impact
Environmental Chemistry and Toxicology
on genome integrity of this early development stage, potentially affecting
eels’ condition and ultimately the population sustainability.
Eutrophication, water quality and nutrient loading: blooms of green algae.
X. Álvarez1; E. Valero1; J. Picos1
1CESAM e Departamento de Química, Universidade de Aveiro; 2CESAM e Departamento de Biologia,
Universidade de Aveiro
The eutrophication in A Baxe reservoir (Galicia, NW Spain) due to the excessive
anthropogenic nutrient loading has promoted the growth of green algae, especially
the harmful cyanobacterial blooms. These blooms threaten the supply of drinking
water for more than 110,000 people, because these occur more frequently with toxic
species, such as the Microcystis aeruginosa.
Surface water samples were collected from one station located next to the
dam, and another at the end of the reservoir. At these points, the physicochemical
measurements were assessed to examine the water quality and chlorophyll-a and
cyanobacteria levels were analyzed in order to determinate the eutrophication.
Diffuse pollution was analyzed from data of livestock farms (pigs, chickens and
cattle) located in the Basin of Umia River, estimating nitrogen and total phosphorus
generated.
Statistical comparisons of water quality demonstrate relationships between
green algae concentrations (Microcystis aeruginosa and chlorophyll-a) and
physicochemistry, especially with temperature and nutrients (total nitrogen
and phosphorus). The nitrogen and phosphorus concentrations influenced the
proliferation of cyanobacteria, therefore the blooms of green algae were associated
with nutrient sources loading as well as the year, in the latter case is due to more
favorable weather conditions in some years than others.
In the study area, there is a lot of agricultural and livestock influence, as a source
of diffuse pollution contributes large amounts of nutrients to aquatic ecosystems.
181
It is one of the biggest problems of eutrophication in the area, which with proper
Environmental Chemistry and Toxicology
management would reduce river pollution significantly. Consequently, this will
improve human health by reducing levels of toxins in drinking water, and will reduce
Composites alto desempeño y cero voc´s para componentes automotrices y
the visual impact that produce blooms each summer.
aeroespaciales, a partir de resina poliéster insaturado dispersable en agua.
Raul Segura Cruz1; Maria Isabel Velazquez Sandoval1; Victor Fermin Gonzalez Saenz1
1AF4 Research Group. Engineering Department of Natural Resources and Environment, Forestry
Engineering College, University of Vigo, Campus A Xunqueira s/n., 36005 Pontevedra, Spain. email:
xaalvarez@uvigo.es. Tel.: +34-986-801-903; Fax: +34-986-801-907.
El objetivo de este desarrollo, es crear materiales compuestos de alto valor
agregado, resultantes de utilizar una nueva resina poliéster dispersable en agua la
cual es única por su capacidad de ser utilizada en proceso de Resin Transfer Molding
(RTM).
Durante la fabricación de materiales compuestos con resinas base solvente, en
etapas de aplicación, vaciado, gelado, curado, etc., se produce emisión Componentes
Orgánicos Volátiles principalmente Monómero de Estireno, lo que afecta al medio
ambiente y en especial al personal expuesto a dichas emisiones por inhalación o
contacto con las mucosas del operador, con graves daños a la salud al ser considerado
como presumible causante de cáncer (National Toxicology Program, report on
carcinogens, 12th edition, 2011), al afectar los fetos en mujeres embarazadas, afectar
al sistema nervioso central y periférico, alteración del sistema endocrino e incluso
afectaciones a hígado y riñones, actualmente existen procesos que llegan a emitir
por evaporación hasta el 14% del monómero de estireno, siendo típico emisiones
del 4 al 6%.
Esta innovación tecnológica permite el desarrollo de materiales compuestos
fabricados a partir de resina poliéster insaturado dispersable en agua, formulada
especialmente para su aplicación en procesos de molde cerrado RTM, que permite
su aplicación para la fabricación de materiales de alto desempeño, con reducción
en el uso y emisiones de Monómero de Estireno a niveles menores al 1% y hasta 20
183
ppm (0.2%) con lo cual son considerados como seguros y reciben la denominación
Environmental Chemistry and Toxicology
de cero o libres de VOC´s, al estar por debajo del 0.5% o 5g/L.
Lo anterior sin afectar las propiedades mecánicas de los materiales finales,
reduciendo costos de materiales y producción.
Las aplicaciones industriales son: Piezas automotrices, para autobuses, de
Aguas residuales como fuente de aporte de piretroides en sedimentos
superficiales y organismos de Bahía Todos Santos, Baja California, México
F.A. Hernández-Guzmán1; J.V. Macías-Zamora2; N. Ramírez-Álvarez2
aeronáutica, embarcaciones; módulos prefabricados para construcción, laminas,
entre otras, sustituyendo gradualmente materiales que actualmente se fabrican con
Resinas Poliéster Insaturadas base solvente.
Los insecticidas piretroides son una familia de compuestos sintéticos utilizados
en la agricultura y en hogares para el control de plagas o vectores transmisores
de enfermedades como el paludismo y malaria. Aunque son de baja toxicidad en
1Grupo Ait - Rase Proyectos Productivos
mamíferos, estudios han sugerido afectaciones a nivel endocrino en organismos
acuáticos. Este trabajo describe el estudio regional realizado en 2013 en el sur de
la Cuenca de las Californias, desde Punta Salsipuedes hasta Bahía Todos Santos, en
el municipio de Ensenada, Baja California, México. Se determinaron por primera
vez diez insecticidas piretroides (bifentrina, fenpropatrina, l-cihalotrina, fipronil
y sus metabolitos, cis/trans-permetrina, ciflutrina esfenvalerato, cipermetrina,
deltametrina) en efluentes de plantas de tratamiento de aguas residuales, sedimentos
superficiales y mejillones (Mytilus californianus). Se colectaron muestras de los tres
principales efluentes de plantas de tratamiento de aguas residuales para caracterizar
los aportes provenientes por esta vía, ya que el aporte por arroyos es escaso debido a la
baja precipitación predominante en la región. Para determinar la distribución espacial
regional y la incorporación por exposición de estos contaminantes en organismos,
se colectaron 32 muestras de sedimento superficial en Bahía Todos Santos y Estero
Punta Banda, así como nueve muestras de mejillones (Mytilus californianus) de tres
localidades. La determinación analítica se realizó mediante cromatografía de gases
con detector selectivo de masas-masas en modo de Ionización Química Negativa
(GC-NCI-MS/QQQ). En sedimentos, el total de las muestras analizadas mostraron
185
concentraciones totales de piretroides (∑10Pyr) en el rango de 0.05-2.02 ng/g dw
Cell and Molecular Toxicology
(median=0.27 ng/g dw) dentro del estuario y de 0.08-6.63 ng/g dw (median=0.18
ng/g dw) en Bahía Todos Santos. Los compuestos frecuentemente identificados en
Cytotoxicity of titanium dioxide nanoparticles in fish cell lines
la zona de estudio fueron deltametrina, cipermetrina, esfenvalerato, permetrina y
A. Bermejo-Nogales1; M.L. Fernández-Cruz1; J.M. Navas1*
bifentrina, cuyas concentraciones son hasta cuatro veces menores a las reportadas
en otros estudios. Se presentan resultados en organismos e interpretaciones de
fuentes y tendencias espaciales.
1Instituto de Investigaciones Oceanológicas/Facultad de Ciencias Marinas - UABC;
2Instituto de Investigaciones Oceanológicas - UABC
Titanium dioxide nanoparticles (TiO2-NP) have a wide number of applications
due to their photocatalyst and ultraviolet blocking properties in cosmetic, solar
and paint industries. The continuous increase in the production of TiO2-NP enhance
the risk for this manufactured nanomaterial (MN) to enter water bodies through
treated effluents or in agricultural amendments. Exposure to TiO2-NP have no
toxic effects in diverse aquatic organisms although there are no conclusive data
on mechanisms of action. Fish cells maintained in vitro are a useful tool to obtain
information about cytotoxic mechanisms of action of a wide variety of substances,
including nanomaterials. The aim of the present study was to assess the toxicity of
three types of TiO2-NP, rutile hydrophobic (NM-103), rutile hydrophilic (NM-104) and
rutile-anatase (NM-105), obtained from the JRC repository using two fish cell lines as
in vitro models: the topminnow fish (Poeciliopsis lucida) hepatoma cell line (PLHC-1)
and the rainbow trout (Oncorhynchus mykiss) fibroblast-like gonadal cell line (RTG2). Dynamic light scattering was used to determine size distributions of dispersions.
Cytotoxicity was evaluated in a concentration range (2 - 256 μg mL-1) after 24 h of
exposure with alamarBlue, CFDA-AM and neutral red assays. TiO2-NP dispersions
showed similar degree of aggregation in both cell culture mediums with z-average
diameter in the range of 225 – 282 nm although NM-103 in PLHC-1 culture medium
show higher polydispersity index and z-average diameter. Disruption of metabolic
activity, plasma membrane integrity and lysosome function was not detected by
exposure to any TiO2-NP at the time and concentration range tested in PLHC-1 and
187
RTG-2 fish cell lines. These results indicate that the use of fish cell lines as in vitro
Cell and Molecular Toxicology
models can facilitate further testing in the framework of integrated testing strategies.
DNA damage and marine pollutants: mechanisms of toxicity, applications
Acknowledgements
This work was financially supported by FP7 MARINA project (263215).
1Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Departamento de Medio
Ambiente, Carretera de la Coruña, Km 7.5, 28040 Madrid, Spain.
E-mail contact*: jmnavas@inia.es
and handicaps of surveying genotoxicity in vivo in non-model organisms
Pedro M. Costa1; Marta Martins1; Maria H. Costa1
Genomic instability is one of the main drivers of loss-of-fitness of wildlife
populations in contaminated areas, due to its acknowledged link to basal metabolic
dysregulation, reproductive impairment and neoplasia-related disease. Polycyclic
aromatic hydrocarbons, dioxins and even some endocrine disruptors are examples
of known organic genotoxicants. These substances’ toxicological pathway may
involve activation through cytochrome P450 monooxygenases (during phase I of
detoxification), generating metabolites that can form bulky adducts with DNA. Metals
exert their genotoxic effects more indirectly, by generating ROS on their own (if
Fenton metals) or by hindering anti-oxidant defences. On the other hand, metals like
Ni, Cd and Hg are also known to impair DNA repair, becoming potent co-mutagens.
The determination of genotoxicity in marine organisms, e.g. through the Comet and
Micronucleus assays and variants, has long been considered as a potential biomarker
for Environmental Risk Assessment, although different endpoints may not necessarily
correlate. For the purpose, fish have been considered as suitable targets due to the
expeditiousness of collecting blood for analyses but bivalves have also been earning
their rightful place. Nonetheless, determining precise cause-effect relationships
between pollutants and DNA damage remains challenging. In large, part such
constraints are due to the complexity of natural environments, to the existence of
toxicant mixtures and multiple biotic factors that hinder the interpretation of results,
from species-specific sensitivity (including the ability to bioactivate mutagens and
189
repair DNA), to age and maturation status. Fish, for instance, are acknowledged to
Cell and Molecular Toxicology
possess more efficient phase I and II detoxification systems whereas DNA repair in
invertebrates is poorly known. Still, the current prospects for the intercalibration
Salinity effect on transcription levels in oysters Crassostrea brasiliana
and standardization of protocols, together with modifications of the Comet assay,
exposed to phenanthrene
for instance, to detect specific types of DNA damage, is leading towards a new level
Flávia Lucena Zacchi1; Daína de Lima1; Fabrício Flores Nunes1; Jaco Joaquim Mattos1; Karim Hahn
Luchmann2; Carlos Henrique Araújo de Miranda Gomes1; Márcia Caruso Bícego3; Satie Taniguchi3;
Sílvio Tarou Sasaki3; Afonso Celso Dias Bainy1
of mechanistic research and biomonitoring, even when non-model organisms are at
stake.
Euryhaline organisms, such as the Brazilian mangrove oyster Crassostrea brasiliana,
1MARE - Marine and Environmental Sciences Centre, Departamento de Ciências e Engenharia do Ambiente,
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa, 2829-516 Caparica, Portugal
possess physiological mechanisms of adaptation to tolerate wide fluctuations in
salinity. These ecosystems are under a constant pressure by the xenobiotics input,
including polycyclic aromatic hydrocarbons, as phenanthrene (PHE). In order to
understand the influence of salinity in the molecular responses of oysters exposed to
PHE, C. brasiliana were acclimatized to different salinities (35, 25 and 10) for 15 days
and then exposed to 100 µg.L-1 of PHE for 24h and 96h. Control groups were kept in
the same salinities without PHE addition. Oysters were sampled for chemical analysis
and gills were removed from gene transcription quantification by quantitative
reverse transcription PCR. It was analyzed the transcription levels of genes from:
oxidative stress pathways (3); xenobiotic biotransformation phase I (3) and phase
II (4); amino acids metabolism (4); fatty acid binding protein (1), osmoregulation
mechanism (1) and aryl hydrocarbon receptor nuclear translocator putative gene
(1). Seven house-keeping genes were tested and only two could be used to data
normalization (Ribosomal_40s_s3-like and Ribosomal_40s_s9-like). High transcript
levels were observed in SULT-like at 24h and 96h; CYP2AU1-like, CYP2-like1 (24h) and
CYP2-like2 (96h) in oysters exposed to PHE at salinity 10 compared to the control
group. These results suggest an increased requirement of PHE detoxification under
hyposmotic stress. ElevatedROS production under hyposmotic stress could explain
191
the higher transcript levels of Catalase-like, SOD-like, GSTmicrosomal-like (96h)and
Cell and Molecular Toxicology
GSTomega-like (24h) in salinity 10 related to salinities 25 and/or 35. The expression of
these genes was not affected by PHE exposure. The amino acid metabolism-related
Exposure to phenanthrene and depuration: changes on gene transcription,
genes (Glutamate_descarboxylase-like (24h), Glycine_transporter-like, Arginase-like
enzymatic activity and lipoperoxidation in gill of scallops Nodipecten nodosus
(96h) and Taurine_transporter-like) also differed between higher and lower salinities,
Romi Sharon Piazza1; Jaco Joaquim Mattos1; Fabrício Flores Nunes1; Daína de Lima1; Flávia
Lucena Zacchi1; Rafael Trevisan1; Álvaro Cavaler Pessoa de Mello1; Miguel Angel Saldaña Serrano1;
Isis Mayna Martins dos Reis1; Carlos Henrique Araújo de Miranda Gomes1; Marisa Bercht1; Claudio
Manoel Rodrigues de Melo1; Afonso Celso Dias Bainy1; C. E. Piazza1
confirming their important role for oyster salinity adaptation, adjusting intracellular
osmolarity.
1Universidade Federal de Santa Catarina;
2Universidade do Estado de Santa Catarina;
3Universidade de São Paulo
Phenanthrene (PHE) is one of the most abundant polycyclic aromatic
hydrocarbons (PAHs) in the marine environment; it is readily bioavailable
and thereby toxic to marine animals. Understanding the mechanism of PHE
biotransformation and cellular responses in bivalves is important to elucidate
the risks of PHE exposure to aquatic organisms. Transcriptional levels of six
biotransformation genes cytochrome P450 (3), glutathione S-transferase (1)
and sulfotransferase (2) were analyzed in gill of scallops N. nodosus exposed
for 24h and 96h to 50 µgL-1 and 200 µgL-1 PHE and depurated in clean water
for 96h. Likewise, the activity of catalase (CAT), glutathione peroxidase (GPx),
superoxide dismutase (SOD), glutathione reductase (GR), glucose 6-phosphate
dehydrogenase (G6PDH), glutathione S-transferase (GST) and the levels of
lipoperoxidation (MDA) were analyzed in this tissue. The organisms exposed
to 200 µgL-1 PHE for 24 and 96 h showed higher transcription levels of CYP2UI,
CYP3A11, GST_HOMOL-like, SULT1B1-like genes. Induction of these Phase I and II
biotransformation genes during the exposure period is possibly associated to
the 2.7-fold decreased levels of PHE in the tissues of scallops submitted to 96h
depuration. Interestingly, the scallops exposed to PHE (200 µgL-1) and further
depurated showed lower SOD and higher CAT activities, which reflected in
193
the lower levels of MDA, possibly related to the PHE depuration. These data
Cell and Molecular Toxicology
indicate that N. nodosus is able to depurate PHE and that possibly this process
involves the participation of Phase I and II biotransformation systems.
Developmental impairment following chronic exposure to ketamine in
early zebrafish (Danio rerio) embryos
1Universidade Federal de Santa Catarina
Luís Félix1; Ana Vidal1; Luís Antunes2; Ana Maria Coimbra3
Ketamine is widely used as an anaesthetic and analgesic by non-competitive
antagonism of the N-methyl-D-aspartate (NMDA) receptors and concerns have
been raised regarding its toxicological and adverse effects. Nowadays, ketamine is
also a club drug and has been classified as an emerging water contaminant. In the
aquatic environment, ketamine may affect aquatic organisms inducing persistent
environmental problems. Previously, we showed that a short (20 min) exposure
to ketamine has a teratogenic potential in early zebrafish embryos and thus, the
present study aimed to evaluate the effects of a longer (24h) exposure on zebrafish
development.
Zebrafish embryos were exposed during 24 hours, (from 2-26 hpf ) to 50, 70 or 90
ng/mL of ketamine based on the calculated lethal concentration (LC50). Six exposure
replicates were performed (100 embryos/group). Developmental disorders, lethality
and/or developmental anomalies were measured at 8, 24, 48, 72 and 144 hpf.
Concentration–responses of general morphology score were also evaluated.Cell
death, using acridine orange, and reactive oxygen species (ROS) generation using
dichlorofluorescin were also assessed.
Ketamine exposure induced several developmental abnormalities, such as:
pericardial oedema, spinal and vertebral anomalies in a concentration-dependent
manner. Also, it decreased zebrafish embryo survival rate. Furthermore, a higher
number of apoptotic cells, mainly accumulated in the brain, heart, and tail regions of
14 a 17 julho 2015
exposed animals were observed, whileno significant changes were observed in the
generation of ROS.
These results reinforce the teratogenic potential of ketamine, which could be an
important issue to aquatic species. Noteworthy, the concentrations of ketamine used
in the present study are 1000-fold greater than the range of concentrations detected
in the environment and thus ketamine real hazard to early-life stages of aquatic
animals will only be clarified after testing environmental relevant concentrations.
1Life Sciences and Environmental School (ECVA), University of Trás-os-Montes and Alto Douro /UTAD),
Quinta de Prados, 5000-801 Vila Real, Portugal;
2Institute for Research and Innovation in Health (I3S), Laboratory Animal Science (LAS), Institute of Molecular
and Cell Biology (IBMC), University of Porto, Rua do Campo Alegre, nº 823, 4150-180 Porto, Portugal;
3Centre for the Research and Technology of Agro-Environmental and Biological Sci-ences (CITAB), Life
Sciences and Environmental School (ECVA), University of Trás-os-Montes and Alto Douro (UTAD), Quinta
de Prados, 5000-801 Vila Real, Portugal
CI CTA 2015
10th Iberian and 7th Iberoamerican Congress
on Environmental Contamination and Toxicology
WEDNESDAY 15.07.2015
BIOMARKERS - AULA MAGNA P0
Ana-Lourdes Oropesa Jiménez In vivo and In vitro effects of anticholinesterase agents on acetylcholinesterase activities of different
rabbit tissues
Juan L. Santos Bitter orange tree leaves as biomarkers of contamination by emerging industrial pollutants in Seville city (Spain)
Mª del Prado Míguez Santiyán Oxidative stress biomarkers in liver and kidney of yelow-legged gulls
Mª del Prado Míguez Santiyán Organochlorine pesticides in fat tissue of wild boar from Galicia (NW Spain): influence of age and gender
Fernanda Dias de Moraes Investigation of b-esterases in the freshwater fish matrinxã (Brycon amazonicus) exposed to cyperme-
thrin based-insecticide
Sebastian Vadillo Gonzaklez Effect of cadium sublethal exposure on cellular redox balance and oxygen consumption in the axolotl Ambysto-
ma mexicanum
Tamara Shalakhmetova The study of biomarkers of toxic effects of oil in the background species of animals from oil con-
taminated territories
Carolina Machado Malheiro Rodrigues Can biochemical tools in benthic macroinvertebrates predict ecological water quality of rivers?
Esther Blanco-Rayón Gender-dependent variability in lysosomal responses and stress on stress of mussel Mytilus galloprovincialis
Mª del Prado Míguez Santiyán A study on the iberian hare as bioindicator of environmental pollution
Mário Araújo The use of Solea senegalensis Kaupt, 1858 as an alternative model to vertebrate animal testing: effects
of a uv-filter to marine fish embryos
Patrícia Silva Lactuca sativa L. as a suitable crop for aluminum toxicity studies
Urtzi Izagirre Sediment health assessment of three estuaries of the bay of Biscay using cell and tissue level bio-
markers in the polychaeta Nereis diversicolor
Diego Ortiz Cañete Temperature compensation in oxidative stress and biotransformation enzyme activities in Mytilus
galloprovincialis exposed to cypermethrin
Antonio Ortiz-Herreros Biomonitoring of chemical pollution in rivers using biomarkers in invertebrates from 2005
to 2014: a review
Amparo Torreblanca Hepatic and gill biomarker responses in Solea spp. From the NW mediterranean fishing grounds
Luis Russo Vieira Influence of temperature on the toxicity of microplastics and Hg to the Mytilus galloprovincialis
Arnaldo Cechinel Bittencourt Transcriptional levels of the gene encoding sulfotransferase in the oyster Crassostrea brasiliana exposed
In situ at Itacorubi mangrove in Santa Catarina, Brazil
Soledad Andrea Chamorro Rodríguez Evidencia de efectos subletales en Daphnia magna expuesta a sucesivos shock tóxicos de licor negro
AQUATIC TOXICOLOGY - AULA MAGNA P1
Ana Leticia Madeira Sanches Toxicological effects of agricultural products kraft 36ec® and score 250ec® and its active ingredients
(abamectin and difenoconazole) in Danio rerio
Julio A. Camargo Toxic effects of fluoride (Fˉ) on the survival and behavior of the freshwater snail Physella acuta
(Draparnaud, 1805)
Mónica Aquilino Amez Evaluation of a endocrine disruptor (vinclozolin) in embryos and adults of Physa acuta
(Gastropoda Pulmonata).
Eugenia López-López Respuesta de la actividad mitocondrial en esporas de Cyathea costaricensis para la evaluación de la
calidad del agua del río Bobos, México
Mónica Aquilino Amez Evaluation of the toxic effects of phthalates on natural populations of Chironomus riparius (Dip-
tera): implications for ecotoxicity studies
Cristina B. García Evaluation of ecotoxicity of two solvents: glycerol derived solvent vs ionic liquid
Soledad Andrea Chamorro Rodríguez Evidencia de efectos subletales en Daphnia magna expuesta a sucesivos shock tóxicos de licor negro
Izaskun Zorita Seasonal variation of micronuclei frequency in common sole of the basque coast (SE Bay of Biscay)
Cleoni dos Santos Carvalho Zinc, copper and cadmium accumulation in liver, kidney and muscle tissues of Lithobates catesbeianus tadpoles
Ana Carolina Faria Costa Transcriptional level of genes involved in the neurotransmitter system of Dicentrarchrus labrax in
response to chronic exposure to pharmaceuticals
Isabel Alves-Pereira Atrazine increases mitochondrial glutathione peroxidase activity of gills of sea lamprey
juveniles during salt acclimation
Nelson Castro Microplastics in fish species of three portuguese estuaries
Marta Martins An application of fish gill histopathology to address the interaction effects between different poly-
cyclic aromatic hydrocarbons (PAHs)
Nicole Colin Assessing the seasonal variability of biotic integrity indices in a mediterranean stream exposed to
industrial sewage discharge: a multi-taxa approach
Marisa Narciso Fernandes Effects of crude extract of cyanobacteria (Radiocystis fernandoi) on the kidney of traíra
(Hoplias malabaricus): oxidative damage and histophatology
Lúcia Guilhermino Effects of temperature and emerging contaminants of high concern on juveniles of the common
goby (Pomatoschistus microps)
Julian Blasco Effect of adsorbed TIO nano-particles on aquatic organisms
2
Sarah Sampaio Py-Daniel Mercury bioaccumulation in piscivorous fishes of an Amazon reservoir, Brazil
Patricia Morcillo Usefulness of fish head-kidney leucocytes to assess the immunotoxicological effects of contaminants
Miriam Hampel Enantiospecific chronic effects of ibuprofen in the freshwater crustacean, A. desmarestii
GLOBAL CHANGES - AULA MAGNA P0
João A. Santos O impacto das alterações climáticas na viticultura europeia segundo os novos cenários do IPCC
Julián Blasco Moreno Biological responses on C. maenas caused by sediment properties and pH interactions during CO2
leakage events from sub-seabed injection and storage
Mariana Hristova Reclamation and quality of technosols in Bulgaria
Fátima Brandão Effects of mercury bioaccumulation in gills of wild fish (Liza aurata) assessed by 1h NMR
metabolomics and oxidative stress endpoints
E.V. Borvinskaya Fish response to the elevated potassium level: the model of the mining area contamination.
Alfredo Pérez Morales Annual cycle of the microzooplanktonic aloricate and loricate ciliates abundances in a
eutrophic lagoon system of the central gulf of California, México
SOIL POLLUTION, CONSERVATION AND RESTORATION - ED. GEO P0
Joana Paula Machado Ribeiro PAHs in soils affected by forest fires in Serra do Caramulo (central Portugal)
Héctor M. Conesa How does metal(loid) pollution affect soil enzymatic activities at metal(loid) enriched mine tailings? The
AQUACULTURE AND ENVIRONMENT - ED. GEO P0
Ana Julia F. Cardoso de Oliveira Microbiological contamination of Sargassum sp from north coast of São Paulo state (Brazil) by
resistant bacteria
role of pioneer plants rhizospheres
Manoel Lago-Vila Chemical characteristics and toxicity of soils from an abandoned Pb/Zn mine
Manu Soto Effects of multiple environmental stressors (thermal stress and cadmium exposure) at different times
and levels of biological complexity in E. fetida
Paula Pérez-Rodríguez Copper fractionation in high resolution sampled vineyard soils managed under heroic viticulture in
Ribeira Sacra (NW Spain)
Paula Pérez-Rodríguez Total copper in the bulk and rhizosphere soil of wild plants grown in long-term contaminated vine-
yard soils from D.O. Ribeira Sacra (NW Spain)
Ana Julia F. Cardoso de Oliveira Microbial loop structure and occurrence of heavy metal resistant bacteria in sediments of Araça
bay (Brazil): influence of contaminants
Paula Pérez-Rodríguez Movilización de cobre por salpicadura de la lluvia en suelos de cultivo de vid
Paula Pérez-Rodríguez Particle size characterization of throughfall from vine leaves sprayed with Cu-based fungicides, by
DLS, AF4-MALS and SP-ICP-MS
Laura Cutillas-Barreiro Study of the competitive release of heavy metals (Cu, Zn, Ni, Cd and Pb) in a copper mine soil
treated with ground mussel shell
Laura Cutillas-Barreiro Mercury distribution among soil aggregate size fractions in spodic horizons of temperate forest
podzols from Galicia (NW Spain)
Laura Cutillas-Barreiro Distribution and accumulation of total Hg in high vertical resolution sampled temperate forest
podzols from Galicia (NW Spain)
Laura Cutillas-Barreiro Contenido de mercurio en suelos y sedimentos de cuencas hídricas afectadas por la actividad
antrópica (Ushuaia, Tierra del Fuego, Argentina)
Xana Álvarez Evaluation of riparian vegetation and invasive species in galician river ecosystems
Lisa Martins Contamination of waters, stream sediments, soils and plants from the abandoned Murçós W-SN
mine area (NE Portugal)
ENVIRONMENTAL CHEMISTRY MONITORING - ED. GEO P1
Viviane Moschini-Carlos Distributions of nutrients and metals in a sediment core and in superficial sediments from a
reservoir used for public water supply São Paulo, Brazil
Maria Antónia Salgado Metal and PAH assessment of the coastline and estuaries in the north of Portugal
Marcelo Pompêo The implementation of an abiotic typology for brazilian reservoirs
Maria João Tomé Rocha Global amounts of polychlorinated biphenyls (PCBs) in the Porto coastline and Douro river estuary
Maria João Tomé Rocha Total amounts of polycyclic aromatic hydrocarbons (PAHs) in the Oporto district
Maria João Tomé Rocha Spatial and annual distribution of estrogenic endocrine compounds in the Aveiro Lagoon
Maria João Tomé Rocha Global amounts of pesticides and their toxic impacts in the Mondego river estuary
Júlio C. O. Lopes Evolução do perfil físico-químico dos sedimentos do estuário do rio Lima
Júlio C. O. Lopes Evolução do perfil físico-químico dos sedimentos do estuário do rio Cávado
Júlio C. O. Lopes Caracterização físico-química dos sedimentos do estuário do rio Minho
Jacinto Elías Sedeño Díaz Diseño de un índice regional de calidad del agua para embalses basado en la lógica difusa
Lisa Martins Natural radiation and geochemical data of the Vila Pouca de Aguiar massif, northern Portugal
Amílcar Teixeira Monitoring the impacts on the ecosystem integrity of Portelo stream (Douro basin, NE Portugal)
after a large spill of mining wastes
Esther Bautista-Chamizo Sediment quality assessment related to benthic macrofauna in São Francisco river (Minas Gerais, Brazil)
J. Arellano Estudios preliminares de evaluación de toxicidad del fenitrotión en dorada (Sparus aurata)
Ana Katsarova Exploring the potential of the zeolite in contaminated soil
ECOSYSTEM LEVEL EFFECTS - ED. GEO P1
Jose Luis Gama Flores Diversidad y dinámica planctónica (fitoplancton y rotíferos) y la producción primaria de un lago
urbano hipertrófico de la Ciudad de México
Alfredo Pérez-Morales Eutrophication due to sewage discharges causes blooms of Peridinium Quinquecorne (Dinophy-
ceae) off the coasts of the Port of Veracruz, Mexico: a case study 2010-2014
NANOTOXICOLOGY - AULA MAGNA P1
Paula Alvarenga
FRIDAY 17.07.2015
Thalita Grando Rauen
Daniela Correia
E M ERGI NG TOXIC C OMPOUNDS - AULA MAGNA P0
Cristina B. García Ecotoxicity of diltiazem
Natalia Ros Ecotoxicology study of a mixture of drugs
Ana-Lourdes Oropesa Jiménez Chronic effects in crustacean Daphnia magna after exposure to a solution of carbamazepine under
single catalytic (TIO2) ozonation
Marta Seoane Physiological alterations on the marine microalga Tetraselmis suecica exposed to the uv filter
benzophenone-3
Sandrine Pontes Machado Effects of psychopharmaceuticals exposure on RNA/DNA ratios in fish species
Bruno Pinto Effects of antidepressant fluoxetine in european sea bass juvenile´s behavior
Larraitz Garmendia Polystyrene microplastic localization and distribution in the gills and digestive gland of mussel
Mytilus galloprovincialis
Alexandre Campos Impact of microcystin contaminated irrigation water on the physiology and mineral content of
carrot (Daucus carota) – implications in crop quality
Effects of silver nanoparticles on different soil exoenzymes activities
Ecotoxicological comparison between industrial waste-recovered and commercial iron oxide
nanocatalysts used in ozonation process
Quantitative distribution and localization of nanoparticle-derived copper in whole cells of Saccharomyces cerevisiae
Silvia Lopes
Joint effects of nanoparticles and respective ionic counterparts to Daphnia magna
Ana Gavina
Impact of an organic nanomaterial on soil invertebrates: SDS/DDAB nanovesicles
Isabel Alves-Pereira
Marisa Narciso Fernandes
Miriam Hampel
Maria Amparo Ochoa Zapater
Titanium dioxide nanoparticles exposure under heat shock conditions caused a partial recovery of
respiratory metabolism of Saccharomyces cerevisiae
Toxicity of TIO2 nanoparticles on the gills of neotropical fish, Prochilodus lineatus: bioaccumulation, oxidative stress and oxidative damage
Itraq-based proteomics analysis of citrate gold nanoparticle exposure effects in the marine clam
Ruditapes phlippinarum
Toxicity studies of insecticide-functionalized mesoporous silica nanoparticles in the milkweed bug
Oncopeltus fasciatus
Irene Biagioni
Effects of mesoporous silica nanoparticles on food intake and reproduction of Blattella germanica
Antonio Ortiz-Herreros
Effects of ingested alumina nanoparticles (AL2O3) in the development and reproduction of Blatella
germanica
Tatiana Andreani
Nanoparticles toxicity studies on microalgae R. subcapitata
Inmaculada Varó Vaello Effect of ibuprofen and carbamazepine on cyclooxygenase (COX-2) gill gene expression, lipids and
reproductive hormones in temperature conditioned sole
Lúcia Guilhermino Influence of temperature on the toxicity of the antibiotic florfenicol to species representative of
freshwater phytoplankton and zooplankton
André M. P. T. Pereira Assessment of the pharmaceuticals removal by different wastewater treatments
Iolanda Ribeiro Potential of constructed wetlands for the removal of antibiotic resistant bacteria from livestock
wastewater
BIOREMEDIATION - AULA MAGNA P1
Ana Filipa Benedito Assunção
PGMS removal from aqueous media using anaerobic bacterial communities from WWTP sludge
Ana Julia F. Cardoso de Oliveira
Diversity and densities of bacteria, cyanobacteria and yeasts from waters and sediments of estuarine areas impacted by hydrocarbon in southeast coast
Tânia Cristina da Luz Palma
Cristiano Soares
CELL AND MOLECULAR TOXICOLOGY - AULA MAGNA P0
Anna V. Lovinskaya Genotoxic effects of rocket propellant’s component on rodents
Tânia Vieira Madureira Interference of estrogenic and androgenic inputs in the brown trout lipid metabolism – experimental
Diogo A. M. Alexandrino
Paulo J. C. Favas
Resistance and bioremoval of paracetamol by a sulphate-reducing bacteria community
Effect of 24-epibrassinolide on ROS content, antioxidant enzymes and Ni accumulation in Solanum nigrum L. under Ni stress
Studies on the biodegradability of two environmentally relevant aliphatic organofluorines
Phytofiltration of uranium-contaminated waters in laboratory conditions
biochemical and molecular studies
Elisabete Santos Secale sylvestre: a wild rye potentially useful for aluminum tolerance study
Cindy Marlene Mesquita Serafim Real time PCR analysis of NMDA receptor expression in zebrafish embryos exposed to ketamine
Ana Vidal Short term exposure to ketamine and its oxidative stress in zebrafish development
DISTRIBUTION OF POLLUTANTS IN THE BIOSPHERE - AULA MAGNA P1
Isabel Párraga Aguado Metal(loid) allocation in the soil-plant system of a Pinus halepensis population growing on
semiarid mine tailings
Víctor M. León Levels and spatial distribution of organochlorinated compounds and polycyclic aromatic
hydrocarbons in fishes from a mediterranean coastal lagoon
MICROBIAL ECOTOXICOLOGY - AULA MAGNA P0
Cátia Venâncio Evidences of salt stress on basidiomycete and zygomycete fungi
Jose Luis Gama-Flores Impacto del cadmio en la actividad nitrificante del bacterioplancton de los canales de Xochimilco
(México): estudio en mesocosmos
Víctor M. León Vertical distribution of PAHs in marine sediments (particulate fraction and interstitial water)
from eight iberian mediterranean areas
Paulo J. C. Favas In-situ phytoextraction of nickel by a native population of Alyssum serpyllifolium on ultramafic sites
(Portugal): prospects for agromining
Paulo J. C. Favas Uranium uptake by native terrestrial and aquatic plants in an uranium mine (central Portugal)
ENVIRONMENTAL CHEMISTRY AND TOXICOLOGY - ED. GEO P0
María Alexandra Bozzo de Brum Repercusión de los plaguicidas utilizados en horticultura protegida sobre el perfil tiroideo y
la colinesterasa plasmática (Salto-Uruguay)
Francisco Soto-Mancera Estudio de los efectos del azufre y del caolín sobre Drosophila suzukii
Víctor M. León Biliary PAH metabolites in european eel (Anguilla Anguilla) from Mar Menor lagoon (SE Spain)
Maria Olímpia de Oliveira Rezende Extração assistida por micro-ondas de um bioerbicida a partir das folhas da Canavalia ensiformis no
combate da erva daninha Emilia sonchifolia na cultura de soja
Angelina Pena Zearalenone as environmental contaminant: occurrence in broa
Raul Segura Cruz Free phthalate plasticizer DOTP (dioctylterephthalate) from chemical recycling of waste pet
Maria Isabel Velazquez Sandoval Development of novel coating free VOC´s (volatile organic compounds) with high anti corrosion
properties
Paulo S. Tonello Kinetic studies and viability of using peat for removal of Al, Cu and Pb of an effluent of a plastic
recycling company
Francisco Soto-Mancera Estudios preliminares de evaluación de toxicidad del fenitrotión en dorada (Sparus aurata)
RISK ASSESSMENT - ED. GEO P1
Silvia Pierre Irazusta Mutagenic activity and indoor air quality in a occupational environment of a images service
of a public hospital in Brazil
Maria Leônia Pessoa da Silva Metais no corpo aquático do riacho Mussuré e o córrego Mumbaba e o reflexo de sua degradação
na saúde dos ribeirinhos - João Pessoa - Paraíba – Brasil
Patrícia Palma Water treatment by nanofiltration and reverse osmosis: chemical and ecotoxicological efficiency
Ruth Pereira Pesticide mancozeb®: derivation of risk limits for freshwater ecosystems
Dércia Santos Histopathological analysis of livers from barbel (Luciobarbus bocagei), nase (Pseudochondrosto-
ma duriense) and trout (Salmo trutta) from Douro basin
Catarina Diniz Adjusting molecular methods to the detection of toxigenic phytoplankton genotypes in
environmental and preserved samples
Margarida P. Reis Cyanobacteria prevalence and cyanotoxin occurrence in an oligotrophic freshwater reservoir
(Beliche – southern Portugal)
Dércia Santos Gill histopathological evaluation in fish species captured in the Douro river basin
Biomarkers
In vivo and in vitro effects of anticholinesterase agents on acetylcholinesterase activities of different rabbit tissues
Ana-Lourdes Oropesa Jiménez1; Juan Javier Chaves Galeano2; Marcos Pérez López2; Francisco
Soler Rodríguez2
The aim of the in vivo study was to investigate the effect of diazinon, an
organophosphate compound, on acetylcholinesterase (AChE) activity in different
tissues of rabbits. Thus, New Zealand white rabbits were exposed to a single oral dose
of diazinon. The animals were divided into 3 groups, consisting of 8 animals each (4
males and 4 females): a control group (exposed to ethanol and distilled water) and
two experimental groups which received doses of 25 and 125 mg/kg b.w. diazinon
(respectively representing 10 and 50 % LD50 for rabbits, 250 mg/kg; Merck index).
After 30 days of exposure the animals were sacrificed and a necropsy was performed
in order to obtain different tissues with demonstrated cholinesterase activity (brain,
cerebellum, medullaoblongata, liver and muscle). Tissue acetylcholinesterase
activities were determined using the Ellman’s method(Ellman et al., 1961) adapted
ENDOCRINE DISRUPTION - ED. GEO P1
Ana-Lourdes Oropesa Jiménez Chronic toxicity of the antiepileptic drug carbamazepine on crustacean Daphnia magna
MONITORING AND MODELLING - ED. GEO P1
Mercedes Arauzo Sánchez Modelización espacial de la contaminación por nitrato en los acuíferos de la Cuenca Alta
del río Ebro (España) mediante SIG
Mercedes Arauzo Sánchez Modelización espacial de los riesgos asociados a usos del territorio que afectan a la contaminación
por nitrato en la Cuenca Alta del Ebro (España)
João A. Santos Monitorização e integração de fatores ambientais na modelação de castas portuguesas”
Simone Varandas Assessment of freshwater bivalves’ communities in Mira lagoons
to microplate. For each tissue, statistical differences in the AChE activity were not
observed between exposed and control groups for both genders (U Mann-Whitney,
p> 0.05) and the AChE activity in the different tissues was in the order: brain ≈
medullaoblongata > cerebellum > liver > muscle.
The objective of the in vitro study was to determine the sensitivity of brain AChE
activity to selected organophosphate (diazinon) and carbamate (oxamyl) insecticides.
After homogenization of brain tissue from non-exposed rabbits, aliquots of the S9fraction were exposed to 2.5, 5, 10, 20 and 40 mM of each one of the insecticides at
25ºC in the dark. The AChE activity was determined using the cited method after 30
205
min, 9, 24 and 48h of exposure. The ability of the selected insecticides to inhibit the
brain AChE activity was: oxamyl > diazinon. The inhibition of brain AChE activity was
dose and time dependent.
Authors wish to thanks the “Ministerio de Educación y Ciencia” from Spain (CTM2007-60041), the
“Consejería de Empleo, Empresa e Innovación. Gobierno de Extremadura” and the FEDER funds for the
economic support.
Reference
Ellman G, Courtney K, Andres V, Featherstone R (1961) A new and rapid colorimetric determination of
acetylcholinesterase activity. Biochem Pharmacol 7:88–95
Toxicology Area, Science Faculty, University of Extremadura, Badajoz, Spain.; 2Toxicology Area, School of
Veterinary Medicine, University of Extremadura, Cáceres, Spain.
1
Bitter orange tree leaves as biomarkers of contamination by emerging
industrial pollutants in Seville city (Spain)
Juan L. Santos1; Pedro J. Barroso1; Julia Martín1; Irene Aparicio1; Esteban Alonso1
Everyday, thousands of new chemicals are discharged to the atmosphere
worldwide. Some of the source discharges are the volatilization of synthetic
compounds, the use of sprays, air fresheners and cosmetics and combustion of
municipal solid wastes. Analytical control of these compounds is difficult, mainly
due to the complexity of sampling (diffuse sources, meteorology, etc.) and their low
concentrations in the atmosphere. These difficulties can be overcome with the use
of living organisms as biomarkers. These biomarkers can be used not only as passive
collectors of pollutants in the atmosphere, but also provide reliable information to
assess the impact of pollutants on the biota and ecosystems.
Bitter orange tree (Citrus aurantium) is particularly widespread in urban and suburban areas from southern Europe, especially in Italy and Spain. Furthermore, it is
evergreen specie, so its leaves are exposed to air pollutants throughout the year. As a
result, this species can be particularly suitable as biomarker of air pollutants.
In this work, the applicability of Citrus aurantium leaves as biomarker of
contamination by emerging industrial pollutants was evaluated. The studied
compounds were two phenolic compounds (nonylphenol (NP) and bisphenol
A (BPA)), two perfluoroalkylated compounds (perfluorooctane sulfonic acid
(PFOS) and perfluorooctanoic acid (PFOA)) and the brominated flame retardant
hexabromocyclododecane (HBCDD).
Analytical method was based on ultrasonic assisted extraction, clean-up by
solid phase extraction and determination byLC/QqQ-MS. Process efficiency was in
the range between 46% and 110%. Precision values, ​​expressed as relative standard
207
deviation, were lower than 10%. Detection limits were in the range between 0.05 and
Oxidative stress biomarkers in liver and kidney of yelow-legged gulls
1.00µg/kg dry matter. Preliminary results showed the presence of NP, PFOS and PFOA
M.P. Míguez1; J. Vizuete1; D. Hernández-Moreno1; L.E. Fidalgo2; A. López-Beceiro2; M. Pérez-López1
in leaves from bitter orange tree. BPA and HBCDD were not detected. 1Department of Analytical Chemistry, Escuela Politécnica Superior, University of Seville, Seville, E-41011,
Spain
To identify and prevent environmental threats to humans and other animals, there
is an increasing need for reliable, stress biomarkers that detect animal health effects at
an early stage. Estimates of elevated oxidative stress may therefore constitute promising
biomarkers of environmental stress in wild animals. Like all aerobic organism, birds are
subjected to basic oxidative processes which are balanced by a variety of antioxidant
system. In this sense, seabirds are useful as bioindicators of pollution because they have
a wide distribution in ecosystems and they are at the top of the food chain. In this study
we analyzed malondialdehyde (MDA) and reduced glutathione (GSH) levels in liver and
kidney of gulls (Larus michahellis) from Galicia (NW Spain), as biomarkers of oxidative
stress. For this purpose, liver and kidney of 54 gulls were taken and stored at -70ºC until
analysis. After homogenization of the samples they were divided according to gender
and age (chicks and adults). MDA was determined using the technique of Recknagel
et al. (1982) and GSH by the fluorimetric method of Hissin and Hilf (1976). The results
show that mean MDA values in gulls were 120.3 ± 40.65 and 91.37 ± 40.98 nmol/g tissue
and GSH values were 63.07 ± 36.04 nmol/g tissue and 42.80 ± 16.93, in liver and kidney
respectively. With respect to the gender, a similar value of both parameters was observed
in liver as well as in kidney, between males and females. However, age seems to influence
both biomarkers since MDA and GSH levels in both organs were higher in adults than in
chicks. Finally, a statistically significant positive correlation between hepatic MDA and
hepatic GSH (p<0.05) was observed, showing a parallel evolution of both parameters.
1Toxicology Unit. Faculty of Veterinary Medicine. Avda de la Universidad s/n. 10003 Cáceres, Spain.;
2Department of Veterinary Clinical Sciences, Faculty of Veterinary Medicine (USC), 27003 Lugo, (Spain).
209
Organochlorine pesticides in fat tissue of wild board fram Galicia (NW
Spain): influence of age and gender
M.P. Míguez1; R Turkmen2; LE Fidalgo3; A López-Beceiro3; D Hernández-Moreno1; M Pérez-López1; F
Soler1
Wild boars represent useful organisms for biomonitoring purposes, not only
because of their ecological relevance on the ecosystem but also because they are a
very interesting hunting species, thus generating food products which are directly
consumed by humans. The aim of this study was to investigate the presence and to
determine the residue levels of organochlorine pesticides (OCPs) in wild boars from
Galicia (NW Spain); and also to determine whether age or gender had any effects on
the fat tissue concentrations of such environmental contaminants. Perirenal fat tissue
from 250 wild boars was extracted with petroleum ether and cleaned up with sulfuric
acid. The final extracts were analised for OCPs by GC-MS in NCI mode. The highest
OCPs concentrations were observed for ∑DDTs (DDE>>>DDT>DDD), followed by
decreasing concentrations according to HCB> ∑HCHs (mainly γ-HCH)> heptachlor
epoxide> ∑chlordane. There was no a statistically significant effect of age on the fat
tissue concentrations of any of the contaminants. However, when considering the
effect of gender, higher pesticide concentrations were found in male wild boars when
compared to females. Our results demonstrate that persistent organic pollutans such
as OCPs have a relevant incidence in the considered area of Spain, thus rendering
biomonitoring studies concerning those toxic agents of high relevance.
1Toxicology Unit. Faculty of Veterinary Medicine. Avda de la Universidad s/n. 10003 Cáceres, Spain.;
2University of Afyon Kocatepe, Faculty of Veterinary Medicine, Department of Pharmacology and Toxicology,
Afyonkarahisar, Turkey;
3Department of Veterinary Clinical Sciences, Faculty of Veterinary Medicine (USC), 27003 Lugo, Spain.
211
Investigation of b-esterases in the freshwater fish matrinxã (brycon
amazonicus) exposed to cypermethrin based-insecticide
F.D. Moraes1; F.P. Venturini1; P.A. Rossi1; I.M. Avilez1; C.A.S. Pigão1; E.A. Almeida2; G. Moraes1
Cypermethrin is a pyrethroid insecticide used in aquaculture, crops, and to
control biological vectors. Fish are highly sensitive to pyrethroids and the nervous
system and biotransformation mechanisms seem to be the key target of the toxicity.
Cholinesterases are involved in the nervous transmission and carboxylesterases
(CbE) in pyrethroids biotransformation. The aim of this study was investigating the
effect of cypermethrin based-insecticide (Galgotrin®) in the B-esterases of matrinxã
(Brycon amazonicus) exposed to 7.2µg L-1 (20%LC50), 14.4µg L-1(40%LC50), 21.6µg
L-1 (60%LC50) and control (no insecticide) for 96 hours. Acetylcholinesterase (AChE)
was kinetically evaluated in the brain and muscle. Butyrylcholinesterase (BChE) was
evaluated in the plasma. CbE was assessed in the liver, gill, intestine, brain and plasma.
Normality of data was estimated by Kolmogorov-Smirnov test. The differences
among groups were assessed by ANOVA one-way (P<0.05), following StudentNewman-Keuls post-test (P<0.05). Brain AChE activity, and brain and hepatic CbE
did not change in the exposed fish. Branchial, intestinal and plasmatic CbE activities
decreased in the fish exposed to higher concentrations. Plasma BChE activity
decresead in the manner-concentration of insecticide (PPMC -0,74, P<0.0001).
Muscular AChE increased in the exposed fish. Brain esterases did not respond to
contamination likely because this organ was protected by specific mechanisms, as
the insecticide sequester by plasma esterases. Branchial and intestine CbE inhibitions
could be due to direct effect of insecticide in these enzymes either to metabolize
it or to protect target organs. Added to CbE inhibition, no liver CbE induction can
contribute to higher toxicity of pyrethroids to fish, since the CbE can be needed to
213
pyrethroid detoxification. The AChE muscle induction was likely to physiological
Effect of cadium sublethal exposure on cellular redox balance and oxy-
response whereas exposed fish presented slow swimming. Therefore, CbE and BChE
gen consumption in the axolotl Ambystoma mexicanum.
may be involved in the sequestering available pyrethroid and protecting target
Sebastian Vadillo Gonzalez1; Adriana García Ávila2; Irma Rosas Pérez3; Javier Miranda Martín del
Campo4; Sebastián Zúñiga Lagunes1; Cecilia Vanegas Pèrez1
organs. In vitro responses are being investigated to clearify these findings. Financial support: Sao Paulo Research Foundation (FAPESP) process 13/14192-9
The axolotl Ambystoma mexicanum is a neotenic amphibian endemic of the
basin of Mexico valley, nowadays under risk extinction due to heavy environmental
Federal University of São Carlos, Dept of Genetics and Evolution, Lab of Adaptive Biochemistry, Rodovia
Washington Luís, km 235 - SP-310, CEP 13565-905 São Carlos - São Paulo, Brazil;
2
Universidade Estadual Paulista “Julio de Mesquita Filho, Dept of Chemistry and Environmental Sciences,
Campus São José do Rio Preto, Rua Cristóvão Colombro, 2265, Jd Nazareth, CEP 15054000 - São José do
Rio Preto, SP - Brazil
1
deterioration and contamination. Heavy metals contamination has being recognize
as one of the main factors responsible of the reduction of the axolotl populations
since concentrations in their environment exceed the permissible limits for the
protection of aquatic life. Among them, cadmium is one of the metals of major
concern due to its high toxicity including its known neurotoxicity, the induction
of oxidative stress and its competitive action with essential metals affecting their
homeostatic regulation. Thus, the objective of this study was to evaluate the
effect of sublethal exposure to cadmium on the cellular redox balance and oxygen
consumption in A. mexicanum juveniles. Organisms were sublethally expose during
21 days to 20 and 200 µg Cd/L, realistic environmental concentrations; a control
group without metal exposure was consider. At the end of the assays hepatic
oxidative stress biomarkers and axolotl oxygen consumption were measure;
additionally hepatic concentration of Cd, Fe, Zn, Cu and Mn were quantify. Obtained
results demonstrate that regardless the significant liver cadmium bioaccumulation,
essential metals remain unchanged (Fe, Zn, Cu, Mn) denoting the maintenance
of its homeostatic regulation. All the biochemical biomarkers evaluated (total,
reduced and oxidized glutathione; enzymatic activity of catalases and superoxide
dismutase; lipoperoxidation and protein carbonils) demonstrate that organisms are
under a severe oxidative stress. However, hepatic metalothioneins levels suggest
their active participation on the detoxification and cellular protection to cadmium
215
injury. Oxygen consumption results demonstrate that the metabolic aerobic
The study of biomarkers of toxic effects of oil in the background species
amplitude and the scope for metabolic activity were significantly alter due cadmium
of animals from oil contaminated territories
exposure and reflected the oxidative stress conditions of axolotls. The results of this
Tamara Shalakhmetova1; Mariya Suvorova1; Leila Sutuyeva1; Assem Ondassynova1; Karlygash
Mukhatayeva1
study demonstrate that cadmium exerts a severe oxidative stress and affects the
energetic status of A. mexicanum juveniles involving probably metal regulation,
compensation and detoxification processes. The obtained information contributes
to the environmental risk analyses of metals exposure in Xochimilco wetland and to
the future management of the axolotl wild populations.
Intensive development of the oil and petroleum industry inevitably leads to
environmental pollution by oil and oil products. The ecological situation can
worsen considerably if these territories belong to an arid zone. For example, in
Kazakhstan where recently intensive oil production is conducted and in natural
landscapes deserts and semi-deserts prevail, the ecosystems are characterized by
Faculty of Sciences, National Autonomous University of Mexico;
Posgraduate Program in Marine Sciences and Limnology, National Autonomous University of Mexico;
3
Centre of Atmospheric Sciences, National Autonomous University of Mexico;
4
Institute of Physics, National Autonomous University of Mexico
1
2
low resistance to the influence of the human activity. In this regard there is a threat
of degradation of desert ecosystems, the risk of sharp reduction of a biodiversity
and deterioration of public health. To assess the state of the biota of various oil
producing regions of Kazakhstan (Atyrau and Mangistau regions) in the background
species of amphibians, reptiles and mammals (rodents) molecular, cellular and
tissue biomarkers of oxidative stress, of a detoxication and genetic disorders were
identified. In background indicator species: lake frog (Rana ridibunda), green
toad (Bufo viridis), water snake (Natrix tessellata) and the great gerbil (Rhombomys
opimus) from the oil-contaminated biotopes of Atyrau and Mangistau region fatty
degeneration and necrosis hepatocyte shrinkage vascular glomeruli and renal
tubular epithelial degeneration, inflammation, a two-fold increase in oxygen
radicals, lipid hydroperoxide (GPL) and malondialdehyde (MDA), and reduction of
the activity of superoxide dismutase (SOD), catalase (CAT), cytochrome P450 and
glutathione-S-transferase in the serum, liver and kidney homogenates were found.
In the great gerbil (Rhombomys opimus) of oil contaminated habitat, as well as in
laboratory rodents (rats and mice) exposed to crude oil from the fields of the studied
regions, level of somatic cells (bone marrow, blood and liver) with a single-stranded
217
DNA breaks also considerably increased. It was found that the activation of lipid
Can biochemical tools in benthic macroinvertebrates predict ecological
peroxidation, inhibition of xenobiotic detoxification and antioxidant protection, as
water quality of rivers?
well as genetic disorders are associated with significant accumulation of aromatic
C. Rodrigues1; R. Pereira1; N. Vieira1
hydrocarbons (benzo (a) pyrene) in the blood and tissues of the test animals. Of
all the bioindicator species studied, in the conditions of arid climate amphibians: a
The use of macroinvertebrates as bioindicators has been showing to be one of
lake frog (Rana ridibunda) and a green toad (Bufo viridis) are the most sensitive to
the most reliable and cost-effective assessment tools of water and habitat quality
oil-contamination. And the most vulnerable area is the territory of the Mangistau
in stream environments throughout the world. However, macroinvertebrates can
region. The improvement of an environmental situation and the preservation
also be a useful tool for detecting and quantifying the responses to complex forms
of a biodiversity in the studied regions requires decrease in an intensification of
of pollution conveniently reflecting cause/effect relationships and overcoming
exploitation of old oil fields, introduction of modern technologies of oil production
the limitations of using isolated biological indicators. and the use of alternative energy sources.
This study was carried out in two Portuguese rivers, the Âncora and Ferreira
Rivers (41°48’5.6’’N, 8°46’28.57”W and 41°11’15.06’’N, 8°27’25.47’’W, respectively),
which are integrated in protected areas under the Natura Network 2000, being, at
Al-Farabi Kazakh National University, Department of Biodiversity and Bioresources, 71 Al-Farabi Av.,
Almaty, 050040, Republic of Kazakhstan
1
the same time, subject to different types and degrees of anthropogenic pressures.
The main aim of this study was to: i) evaluate the ecological quality of; both
rivers by using tools recommended by the Water Framework Directive (biological
parameters - benthic macroinvertebrate community – as well as physico-chemical
and hydromorphological parameters); ii) analyse several oxidative stress (e.g.
catalase, glutathione peroxidade, glutathione-s-tranferase, lipid peroxidation)
and neurological biomarkers (acetylcholinesterase) in macroinvertebrates most
representative of the existing communities in the studied rivers and, iii) verify if
there is a relationship between the levels of activity of the analysed biomarkers
and the environmental quality of the streams, previously performed.
Sampling of benthic macroinvertebrates for the assessment of the ecological
quality (according to the Directive 2000/60/CE) and for the analysis of biomarkers
took place during October 2015, at different sampling sites of both rivers.
219
The present study will provide new insights about the potential use of
additional and most sensitive parameters to assess water quality of streams.
Gender-dependent variability in lysosomal responses and stress on
stress of mussel Mytilus galloprovincialis
E. Blanco-Rayón1; I. Marigómez1; U. Izagirre1; L. Garmendia1
1Faculty of Sciences of the University of Porto – Biology Department, Campo Alegre Street, 4169-007 Porto, Portugal;
2CIIMAR – Interdisciplinay Centre of Marine and Environmental Research, University of Porto, Bragas
Street, 289, 4050-123 Porto, Portugal.
Mussels are widely used sentinel species for marine ecosystem health
assessment based on biomarkers. Standard procedures are available for the
determination of biomarkers but there is a need to optimize them by reducing
the influence of natural variability and the effects of confounding factors.
In the present study, it was investigated how gender of mussels (Mytilus
galloprovincialis) may affect in digestive gland lysosomal biomarkers and
stress on stress test. Accordingly, mussels were collected from a reference site
(Plentzia) and from a chronically polluted site (Arriluze) in the Basque Coast in
March 2013 and January, April, August and November 2014. Lysosomal structural
changes, intracellular accumulation of neutral lipids and stress-on-stress test
were analyzed, and gamete development stages were determined. Considering
both genders, lysosomal responses were as expected for seasonal and pollution
history. Lysosomal biomarkers were not influence by gender, except in March
2013 when male mussels had larger lysosomes than female mussels in Plentzia.
Intracellular neutral lipid accumulation showed differences between genders
in both localities, mainly related to gamete developmental stages. Stress on
stress test showed a clear trend in Arriluze were female mussels survive longer
than male mussels. In Plentzia, female mussels had significantly shorter survival
than male mussels in April 2014. Overall, lysosomal biomarkers appeared to be
slightly more sensitive in female mussels than in male mussels. Thus, the use of
female mussels could be more suitable than male mussels for the assessment of
marine ecosystem health status. Nevertheless, the observed differences mainly
221
in lysosomal biomarkers were not subtle enough to no compare previously
A study on the Iberian hare as bioindicator of environmental pollution
recorded data even the gender was not determined. There is a lack of knowledge
J. A. González-Moreno1; M.P. Míguez1
in gender-dependent variability on biomarkers, which could be a crucial factor
Recent environmental studies claim that the hare can be a good bioindicator of
for the assessment of the marine ecosystem health status.
environmental pollution because this species has a sedentary lifestyle, is sensitive
to changes in ecosystems and the number of animals hunted every year is high. The
Acknowledgements
This work has been funded by the Ministry of Economy and Competitiveness (MINECO 2013-2015), and
by the Government of the Basque Country through a pre-doc grant to Blanco-Rayón (PRE_2013_1_640).
1
Cell Biology and Environmental Toxicology Research Group; Research Center for Experimental Marine
Biology and Biotechnology (PiE-UPV/EHU), University of Basque Country, Areatza, Plentzia – Bizkaia,
Basque Country, Spain
objective of this study was to establish the physiological levels of oxidative and
nitrosative stress biomarkers as malondialdehyde (MDA), reduced glutathione (GSH) and
nitric oxide (NO) in liver and kidney of Iberian hare. In addition, we sought to determine
the influence of variables such as gender, type of hunting or parasitism. The hares used
(Lepus granatensis) (34 males and 37 females) were obtained from controlled hunts in the
community of Extremadura and Toledo (Spain). The two types of hunting employed were
hunting by shooting (42 hares) (shotgun) and hunting by greyhound (29 hares). Also, all
hares from this study were classified in parasitized and non-parasitized by Cysticercus
spp. (the most common parasite species in hares). Livers and kidneys were collected and
frozen (-80ºC) until start the determinations. In liver, parasitized hares group showed
higher levels of MDA and NO, and lower levels of GSH than non-parasitized group. In
addition, animals hunted by greyhound showed lower levels of GSH than animals
hunted by shooting. Therefore, the presence of parasites and the hunting by greyhound
could induce oxidative and nitrosative stress in liver. In kidney, male hares showed higher
levels of MDA than females. Moreover, animals hunted by shooting showed lower levels
of MDA than animals hunted by greyhound, suggesting that the hunting by greyhound
could produce oxidative stress in this organ. In conclusion, we can consider that the
Iberian hare could be a good bioindicator of environmental pollution, but the type of
hunting, the parasitism and gender should be taken in consideration in these studies.
Toxicology Unit. Faculty of Veterinary Medicine. Avda de la Universidad s/n. 10003 Cáceres, Spain.
1
223
The use of Solea senegalensis Kaup, 1858 as an alternative model to
vertebrate animal testing: effects of a UV-filter to marine fish embryos
M. J. Araújo1; M. S. Monteiro1; R. J. Rocha1; A. M. V. M. Soares1
Organic UV filters, such as 4-methylbenzylidene camphor (4-MBC) are commonly used
in personal care and cosmetic products. The 4-MBC, which is reported to act as endocrine
disruption substance (EDs), is currently found in sediments, water and aquatic biota;
however, its effects on marine species, particularly during early life stages, are largely
unknown. Solea senegalensis is a benthic species with high economic importance that
occurs throughout the Iberian coast. Larval stages arise as a good model for evaluating
the effects of 4-MBC in marine ecosystems. Therefore, our goal was to evaluate the
effects of this substance on mortalitiy, malformations, growth, biochemical biomarkers
and behavior of S. senegalensis early life stages.
Fertilized eggs were obtained from a commercial hatchery (Sea8, Portugal) that use
wild S. senegalensis spawner stocks. Exposure to 4-MBC was performed immediately after
arrival to the laboratory until 96hpf (hours post-fertilization). A first trial was performed to
evaluate malformations and mortality of the fish larvae and for the determination of the
LC10 at 96hpf (0.73±0.078 mg/l). In a second trial embryos were exposed to a sub-lethal
range of 4-MBC below the LC10 and checked daily for mortality, malformations, growth,
lethargy and delayed development. The effects at biochemical level were performed
through the determination of the enzymatic activities of lactate desidrogenase,
catalase, glutathione s-transferase and acetylcholinesterase and the determination lipid
peroxidation as indicator of oxidative damage. Behavioural response to light stimulation
after 96 hpf was also assessed in Zebrabox® equipment (Viewpoint, France).
In a preliminary analysis of data, the exposure to increasing concentrations of 4-MBC
seems to promote a reduction in fish length, an increase of malformations and inducing
225
alterations in larvae behavior.
Results will be discussed integrating the data obtained at the different levels of
Lactuca sativa l. as a suitable crop for aluminum toxicity studies
Patrícia Silva1; Maria João Barros1; Elisabete Santos2; Manuela Matos2
biological organization, aiming at unravelling the main adverse outcomes of 4-MBC
The use wastewater, which may be contaminated with metals, for irrigation in agriculture
in S. senegalensis larvae.
is common all over the world and can increase the concentration of these agents in the
soil, decreasing the crops yield. The main objective of this study, was to investigate the
Acknowledgments
This work was supported by the Portuguese Foundation for Science and Technology (FCT) through
CESAM (UID/AMB/50017/2013) and through the scholarships of MJA (SFRH/BD/52572/2014) and MSM
(SFRH/BPD/45911/2008).
toxicological effects of aluminum (Al) on seed germination and seedling growth of lettuce
Department of Biology & CESAM, University of Aveiro, 3810-193 Aveiro, Portugal
cultivars, referenced in studies with different behavior to the presence of heavy metals, were
1
(Lactuca sativa L.), one of the most cultivated crops worldwide, important for human nutrition
and referred as a standard species for toxicity and genotoxicity tests. Thus, seeds of two lettuce
exposed to five different Al legal concentrations. For each treatment 100 seeds were used
and the number of germinated seeds was counted daily for 7 days, as well as the germination
percentage and root and shoot lengths that were measured at the end of the assay. Also, the
germination rate and the vigor index was calculated, and the proline content was estimated
for all doses tested. Results showed that seed germination was not inhibited by Al, but the
other factors were negatively affected by this metal. We conclude that the use of effluents
or wastewaters contaminated with legal levels of Al as irrigation water can reduce the
production and the quality of crops. Also, this work presents lettuce as an appropriate crop for
toxic studies, as well as several parameters as reliable biomarkers / bioindicators of Al toxicity.
Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, 5000-801, Vila
Real, Portugal;
2
Center of Agricultural and Genomics Biotechnology (CGBA-UTAD), University of Trás-os-Montes and Alto
Douro, 5000-801, Vila Real, Portugal.
1
227
Sediment health assessment of three estuaries of the Bay of Biscay using
cell and tissue level biomarkers in the polychaeta Nereis diversicolor
Olatz Romo1; Urtzi Izagirre1; Beñat Zaldibar1
Estuaries constitute transition zones leading to high biological productivity and
are constantly exposed to important urban and industrial activities responsible for
numerous perturbations. Therefore, species inhabiting sediments are particularly
exposed to chemical stress that may affect their health status. In the present work
sediments from three estuaries in the Basque Country with different pollution levels
(Plentzia, Arriluze and Pasaia) were collected and transported to the laboratory.
Ragworms (Nereis diversicolor) were purchased and let burrowed into the sediments
and then collected after one and seven days of exposure for chemical and biological
measurements including, bioaccumulation of heavy metals, changes in size and
weight and cell and tissue level biomarkers. Although in general sediments presented
low to moderate heavy metal pollution, sediments from Arriluze and Pasaia presented
higher Cr, Cu, Pb and Zn levels than sediments from Plentzia. These higher values
lead in lysosomal and neutral lipid alterations observed in ragworms burrowed in
changes at cellular and subcellular levels in ragworms (lysosomal and neutral lipid
alterations) but not at higher biological organization levels (weight). These results
indicate the potential of Nereis diversicolor as sentinel specie for sediment health
status assessment.
Acknownledgements
Work funded Basque Government through Consolidated Research Groups fellowship (IT810-B).
1University of the Basque Country
229
Temperature compensation in oxidative stress and biotransformation
enzyme activities in Mytilus galloprovincialis exposed to cypermethrin.
Diego Ortiz Cañete1; Inmaculada Varó Vaello2; Amparo Torrebalnca1
In ectothermic organisms, as the mussel Mytilus galloprovincialis, temperature
is recognized as a pervasive factor affecting structures and functions at all levels
of biological organization and can cause alterations in biochemical, cellular
and physiological rates. Metabolic enzyme activities are strongly affected by
temperature, and generally reaction rates are slower at low temperatures.
Ectothermic animals can respond, on both short-term and evolutionary time
scales, to chronic temperature changes by quantitative and/or qualitative
adjustments in enzyme activity. Thermal acclimation of metabolism includes
changes in the concentration of enzymes, maintaining the enzyme´s affinity
for substrate and turnover rates as temperature changes, or the induction of
enzymes with different kinetic properties such as an allozyme with a lower
energy of activation. In order to evaluate biochemical temperature compensation
in catalase (CAT) and Glutathione-S-transferase (GST) activities, mussels were
kept at three temperatures (17ºC, 22ºC and 27 °C) and two cypermethrin
concentrations during 96h following a factorial design. Activities were measured
and compared at two different conditions: at a selected common temperature
of 20ºC for all experimental groups and at the acclimation temperature of each
one (17ºC, 22ºC and 27 °C). When measured at 20ºC the highest CAT activity
was reached at the lowest temperature of acclimation whereas the highest GST
activity was reached at 22ºC. When activities were measured at the temperature
of mussel acclimation, temperature compensation processes were detected for
GST and CAT. Total temperature compensation was found in CAT activity. D. Ortiz
231
is recipient of a fellowship from PhD (Becas Chile) for Advanced Human Capital
Biomonitoring of chemical pollution in rivers using biomarkers in inver-
formation pertaining the Ministry of Education of Chile (CONICYT) and funded by
tebrates from 2005 to 2014: a review
the Government of Chile.
A. Ortiz-Herreros1; J. del Ramo1; A. Torreblanca1
The use of biomarkers has become in recent decades one of the most efficient tools
Universidad de Valencia; 2Instituto Torre la Sal
1
for pollution biomonitoring in aquatic habitats. Vertebrates (mostly fish) and other
groups of animals have been used as sentinel organisms for biomonitoring of chemical
pollution in rivers. However, use of vertebrates for scientific uses has many limitations
because of ethics and current EU regulation. This is the main reason invertebrates are
better candidates for their future use as sentinels organisms. Many questions arise when
analysing the current state of the art: what are the species most frequently used? What
are the biomarkers preferred? How sensitive are biomarkers to the contaminant insult?
Are the studies conducted so far evenly distributed geographically? In order to find an
answer to these questions 46 articles published in the topic from 2005 to 2014 have been
systematically and deeply analysed in order to get quantitative information.
The results revealed that most of the studies were carried out in European countries
like Spain and France, and invertebrates considered invasive species (Procambarus
clarkii, Dreissena polymorpha) are the preferentially chosen as sentinel organisms. The
most commonly used biomarkers in these studies are some enzymatic activities as
cholinesterases, catalase, mixed function oxidases and glutathione transferase. Aspects as
feasibility of biochemical analysis, sampling limitations and funding are the main aspects
considered to explain the results. Finally, future needs for the further development of the use
of biomarkers in invertebrates in the biomonitoring of chemical pollution in rivers is given.
Departamento de Biología Funcional y Antropología Física. Universitat de València. Dr. Moliner 50,
Burjassot, 46100 Valencia, Spain.
1
233
Hepatic and gill biomarker responses in Solea spp. from the NW
mediterranean fishing grounds
Roser Siscar1; Amparo Torreblanca1; Jose Francisco Pertusa1; Montserrat Solé2
The common sole, Solea solea and the Senegalese sole, Solea senegalensis are two
important commercial species that coexist in the NW Mediterranean Sea. Several
common biomarkers of chemical exposure were measured in two organs involved
in biotransformation, although to a different extent: liver and gills. These common
biomarkers were: phase I CYP1A-dependent EROD activity and carboxylesterase, as
phase II reactions GST and the antioxidant defences: catalase, glutathione reductase
and glutathione peroxidase. Histological examination of the liver was assessed as
percentage of (1) piknotic nuclei and (2) lipid vacuoles as a sign of cell damage. Site
differences were more evident in S. senegalensis as individuals differed from two
more distant sites. Principal Component Analysis (PCA) considering all chemical
and biomarker variables discriminated individuals collected at the different fishing
grounds. Histological damage was not observed in relation to sites but, instead, it
was affected by the fish size.
Acknowledgements
DEPURAMAR Project CTM2010-16611.
University of Valencia; 2Institut de Ciencies del Mar (ICM-CSIC)
1
235
Influence of temperature on the toxicity of microplastics and hg to the
Mytilus galloprovincialis
L. R. Vieira1; M. Oliveira2; L. Guilhermino1
Pollution with plastic litter has been recognized as a worldwide problem, with
severe impacts on aquatic ecosystems. During the past years, the risks involving
larger plastic debris to marine ecosystems have been well documented. However,
the available data concerning the effects of microscopic plastic debris is limited and
even less is known regarding microplastics contaminated with important marine
contaminants such as metals. Additionally, in the present context of global warming,
more knowledge on the combined effects induced by temperature changes and
mixtures of relevant environmental contaminants is urgently needed. Therefore, the
main objective of this study was to investigate the effects of temperature increase on
the toxicity of microplastics alone and in mixture with a heavy metal (mercury), to the
Mediterranean mussel, Mytilus galloprovincialis, a species with a wide distribution in
the NW Portuguese coast. Mussels were exposed for 96h to different concentrations
of both contaminants individually and in mixture, at both 20 and 25°C. Several
biomarkers, involved in neurotransmission, energy production, biotransformation
and oxidative stress were analyzed as effect criteria after 96h of exposure and after
192h of recovery in control seawater. Overall results indicated adverse effects of the
selected stressors on mussel’s health, highlighting the need of more research of
potential impacts of plastic-metal interactions in the aquatic environment, crucial to
establish the biological consequences of these emerging contaminants.
University of Porto: ICBAS & CIIMAR: ICBAS-Department of Population studies, Laboratory of Ecotoxicology;
CIIMAR-Research Group of Ecotoxicology, Stress Ecology and Environmental Health, Rua dos Bragas, 289,
4050-123 Porto, Portugal; 2
University of Aveiro: Department of Biology & CESAM, 3810-193 Aveiro, Portugal.
1
237
Transcriptinal levels of the encoding sulfotransferase in the oyster Crassostrea
brasiliana exposed in situ at Itacorubi mangrove in Santa Catarina, Brasil
A. Bittencourt1, J. J. Mattos1, A. L. Dafre1, A. C. D. Bainy1 , M. R. F. Marques1
Coastal marine environments, specially semi-enclosed systems like mangroves, are
highly susceptible to contamination caused by urban and industrial development.
Itacorubi mangrove, located within the city limits of Florianópolis, Santa Catarina, southern
Brazil, has been experiencing constant anthropogenic pressure, mainly by the inflow
of domestic sewage. Renowned touristic destination, Florianópolis population doubles
during summer. We conducted two experiments, during and after the 2011/2012 summer
season, when specimens of C. brasiliana were transplanted to the river that drains Itacorubi
mangrove basin; the reference group was placed at Ratones mangrove, a pristine area.
mRNA was extracted from gill tissue sampled at 24 and 96 hours after in situ exposure
and cDNA was used to asses transcript levels of the gene encoding sulfotransferase (SULTlike) by quantitative real-time PCR (qPCR), levels were calculated using the 2-∆CT method.
Therefore, α-tubulin-like, β-tubulin-like and 28S rRNA-like were selected as reference genes.
After 24 hours, transcription level in summer was heavily induced (28.5 times fold). After
summer, SULT levels were higher relative to the control group at both 24 and 96 hours
(6.8 and 8 fold, respectively), although no statically difference was seen between them.
The increased level of SULT transcripts seen in summer after 24 hours of in situ exposure,
suggests the potential of this gene as biomarker of environment contamination by sewage.
Further experiments to rule out eventual seasonal transcriptional variations of SULT, along
with the analysis of the transcription of other genes related to xenobiotic biotransformation,
may be relevant to validate SULT as biomarker in this mollusk.
1
LABCAI, Aquaculture Department, Federal University of Santa Catarina, Rodovia Admar Gonzaga 1346,
88034-001 Florianópolis, SC, Brazil
239
Global Changes
O impacto das alterações climáticas na viticultura europeia segundo os
novos cenários do IPCC
Helder Fraga1; Aureliano C. Malheiro1; Ricardo Costa1; José Moutinho-Pereira1; João A. Santos1
A vitivinicultura é fortemente dependente do clima, sendo o desenvolvimento e
crescimento da videira influenciado pelas condições atmosféricas. Tendo em conta
a mudança climática projetada, esta cultura poderá ser afetada no futuro. Devido à
importância do sector vitivinícola na Europa, a avaliação dos impactos das alterações
climáticas nesta cultura é de extrema relevância para o setor. O presente estudo foi
realizado através da análise de simulações climáticas, provenientes de um conjunto
de 9 modelos climáticos regionais, com base nos cenários (RCPs; Representative
Concentration Pathways) 4.5 e 8.5 do IPCC (Painel Intergovernamental sobre Mudança
Climática) até 2050. Para avaliar as condições ideais para a viticultura atual e futura,
foram determinados os graus-dia (GDD) sobre a Europa, numa elevada resolução
espacial (<1km). Alterações na média e no sinal climático foram analisadas para
cada cenário futuro. Os resultados mostram que em ambos os cenários futuros, está
projetado um aquecimento significativo sobre toda a Europa, reforçado na RCP8.5.
Sobre o sul da Europa, o aquecimento projetado deverá ter impactos negativos no
desenvolvimento da videira, impondo a necessidade de medidas de adaptação
adicionais para lidar com o stress térmico. Na Europa central, são esperadas alterações
nas condições de desenvolvimento, podendo resultar em mudanças na tipicidade
do vinho. Por outro lado, ao longo do norte da Europa, o clima mais quente pode
resultar em condições mais adequadas para o desenvolvimento da vinha. Com base
nestes resultados, é esperado que a mudança climática traga novos desafios para
241
o sector vitivinícola na Europa. Neste sentido, medidas de adaptação adequadas
Biological responses on C. maenas caused by sediment properties and
devem ser adotadas, a fim de lidar com o impacto negativo das alterações climáticas
ph interactions during CO2 leakage events from sub-seabed injection
na viticultura, especialmente nas regiões do sul da Europa. Nas regiões do centro e
and storage
norte da Europa, apesar do aumento geral da adequação vitivinícola, também são
Araceli Rodriguez Romero1; Natalia Jiménez Tenorio2; Inmaculada Riba López2; Julián Blasco
Moreno1
necessárias medidas de adaptação às novas condições climáticas.
CO2 capture and storage in sub-seabed geological formations is proposed as one
Centre for Research and Technology of Agro-Environment and Biological Sciences (CITAB), Universidade
de Trás-os-Montes e Alto Douro, 5000-801 Vila Real, Portugal
1
of the potential options to mitigate the abrupt and irreversible consequences of
climate change. However, possible CO2 leakages could occur during the injection and
sequestration procedure, provoking significant repercussions on marine environment. In this study, we investigate the toxicity effects of acidification derived from possible
CO2 leakage events on the European green crab, Carcinus maenas. To this end, a labscale experiment involving direct release of CO2 through marine sediment was conducted
at expected scenarios of pH values for this kind of events. Crabs were exposed for 10 days
to sediments collected from two different coastal areas, one with relatively uncontaminated
sediments (RSP) and the other with known contaminated sediments (MZ and ML), under
the pre-established pH conditions (8.0-6.1). End points measured were: survival rate,
histopathological damages and metal accumulation (Fe, Mn, Cu, Zn, As, Cr, Cd and Pb) in gills
and hepatopancreas. Significant associations were observed between pH and the histological
damage. Organic matter and organic carbon content as well as Fe, Mn, Cr, Pb, Cd and PAHs
concentrations in sediment presented significant correlation coefficients with the damage
to gills and hepatopancreas and metal accumulation in both tissues. The results obtained in
this study revealed the importance of sediment properties in the biological effects caused
by possible CO2 leakage events derived from the implementation of this mitigation option.
However, a clear pattern was not observed between metal accumulation in tissues and pH
reduction. Animal´s avoidance behaviour and tolerance degree to acidification may are
confounding factors to assess metal bioaccumulation. Further research to find a suitable
243
organism battery, which allows us to predict the possible consequences on human health
Reclamation and quality of technosols in Bulgaria
derived from metal bioaccumulation associated to high CO2 conditions, is required.
Mariana Hristova1; Ana Katsarova1
Instituto de Ciencias Marinas de Andalucía (ICMAN-CSIC);
Universidad de Cádiz
1
The different ways of mining for mineral resources impacts irreversible damages
2
to the pedosphere. The damages on the landscape from opencast mining are
recovered by law in Bulgaria. The main way is to cover the disturbed landscape with
topsoil rich in humus. The natural soil materials from reclamation have to be no less
than 40 cm. According to this method many areas are recovered and returned to
agricultural use.
There are cases where the lack of surface soil humus, the reclamation is done
without humus layer with suitable by physical and chemical characteristic geological
materials. That type reclamation is based on good scientific basis and it’s observes for
the forestry use.
The paper presents the soil properties of Technosols, designed by reclamation
after activities of the main Bulgarian company for coal mining “Maritza-East”. The
studied are the properties of Technosols by the main method for investigation - the
forming the soil profiles.
Evaluated are the basic soil indexes as pH, sorption capacity, soil organic matter,
the basic nutrients and the level of normed heavy metals. The study concluded
that even though the humus reclamation is the principal method by law, the direct
reclamation may to be use in practice when is based on the good scientific basis.
Institute of Soil Science, Agrotechnologies and Plant Protection “N.Poushkarov”, Sofia, Bulgaria
1
245
Aquatic Toxicology
Toxicological effects of agricultural products kraft 36ec® and score 250ec®
and its active ingredients (abamectin and difenoconazole) in Danio rerio
Ana Leticia Madeira Sanches1; Bruna H. Vieira1; Marina Reghini1; Michiel A. Daam1; Evaldo Luis G. Espíndola1
Abamectin and difenoconazole are actives ingredients used as insecticide
and fungicide, respectively, and the toxic effects of these compounds to nontarget organisms, including aquatic ecosystems, are already noted in others
studies. However, this toxicity is related to the actives ingredients effects. In the
field, the pesticides Kraft (abamectin) and Score (difenoconazole) are extensively
used in several tropical crops, especially in potato and strawberry’s crops, and
toxicological studies about the commercial products effects are requests.
Thereby, the aim of this study was to investigate if commercial products Kraft
and Score, in the concentrations recommended by the manufacturers for the
strawberry’s crops, can be more harmful to non-target organism than actives
ingredients abamectin and difenoconazole in acute exposures. For this, toxicity
tests were done using as test organisms the species of fishes Danio rerio,
following the ABNT (Brazilian Association of Technical Standards) standards.
The mortality after 48 hours of exposure were observed and de LC 50 were
calculated with the TrimmedSpearman Karber method. In general, we observed
that commercial products showed to be more toxic to Danio rerio than actives
ingredients. For abamectin and Kraft the LC 50were 90.8 µg/L and 72.3 µg/L
respectively; showing an increased toxicity in more than 20% of commercial
product in relation to active ingredient. The values of LC50 to Difenoconazole
and Score were 2.1 mg/L and 1.39 mg/L, respectively; and the increase of the
247
toxicity of commercial product in relation to active ingredient was observed
Toxic effects of fluoride (Fˉ) on the survival and behavior of the freshwater
(36%). The results indicate that the commercial products are more dangerous
snail Physella acuta (Draparnaud, 1805)
to aquatic non-target organisms than its actives ingredients. On the other
Julio A. Camargo1; Álvaro Alonso1
hand, the toxicity values provided by the manufacturers in the chemical safety
information sheet only refers to active ingredient effects values, but this
information does not corroborate with the reality. So, we aim to provide data
and prove that more strict laws must be implemented for the pesticide use,
especially in tropical regions.
Departamento de Ciencias de la Vida, Universidad de Alcalá, 28005 Alcalá de
Henares (Madrid), España.
We carried out laboratory experiments to assess the toxic effects of fluoride
(Fˉ) on the survival and behavior of the pulmonate freshwater snail Physella acuta.
Test animals were obtained from an unpolluted reservoir. After acclimatization
for ten days to laboratory conditions, snails were exposed to a range of fluoride
CRHEA - EESC - USP
1
concentrations (50 - 300 mg F–/l), plus a control (0.17 mg F–/l), for four days. Test
fluoride concentrations were made from sodium fluoride (NaF). The static (with water
renovation) short-term toxicity bioassay was conducted in triplicate using 18 glass
vessels and 180 snails (5 juveniles + 5 adults per vessel). Vessels were covered with
perforated plastic surfaces in order to prevent the escape of snails, which were fed ad
libitum with algae. Dead and immobile (but still alive) snails were daily checked.
We also examined the escape response to fluoride by monitoring the proportion
of snails at each treatment that were located either on the water line or above the
water. In addition, just after finishing the toxicity bioassay, all surviving animals were
kept in their respective glass vessels with renewed dechlorinated tap water (but with
no addition of fluoride) during the following seven days. The recovery of snails was
checked by comparing the sliding movement rate (expressed as mm/s) of affected
snails (exposed to fluoride toxicity) with the sliding movement rate of control snails.
Mortality, immobility and the escape response tended to increase with increasing
fluoride concentration and exposure time, juveniles showing higher sensitivity to
fluoride than adults. The recovery of snails tended to increase with time, but full
249
recovery only occurred in affected snails exposed to lower fluoride concentrations.
Evaluation of a endocrine disruptor (Vinclozolin) in embryos and adults
Overall, after comparing the obtained data with previous data for other aquatic
of Physa acuta (Gastropoda pulmonata).
animals, we conclude that P. acuta is a relatively tolerant species to fluoride toxicity.
M. Aquilino1,2, G. Morcillo1, JL. Martínez-Guitarte1, P. Sánchez-Argüello3
Vinclozolin (Vz) is a fungicide which inhibits the androgen receptor (AR) in
1Universidad de Alcalá
vertebrates. This feature, along with the probability that this agrochemical reach
surface waters, justifies the need to study their effects on aquatic organisms. Along
with its known effect as endocrine disruptor, new studies have suggested that Vz to
alters epigenetic mechanisms.
This work analyzes the effect of Vinclozolin at different stages of the life cycle
of Physa acuta, a freshwater Gastropoda. Specifically it has been compared the
adult mortality after chronic (45 days) and acute exposures (96 h) and as well as
reproductive capacity (number of eggmasses) in the presence of Vz. Besides these
tests are complemented with an embryo exposition to determine if Vz effects are
stronger in early stages of Physa acuta development.
Although the highest concentration of Vz (5 mg/L) only produced lethal effects
in chronic exposition, while the number of eggmasses was affected in both cases.
Embryotoxicity analysis indicates that embryos are less sensitive than adults to this
compound. Finally, DNA methylation was analyzed in adults and embryos to test
putative changes in the pattern resulting from exposure to Vz.
This study adds new information about the effects of Vzin Physa acuta, a suitable
gastropod to investigate events mediated by non-nuclear receptors and effects not
associated to endocrine alterations, as epigenetic mechanism, since there are not
functional steroid receptors described in mollusks.
This work was supported by the Plan Nacional de Investigación Científica, Desarrollo
e Innovación Tecnológica (Spain), grant CTM2012-37547 from the Ciencias y Tecnologías
251
Medioambientales program.
Respuesta de la actividad mitocondrial en esporas de cyathea costari-
M.A. is the receiver of a predoctoral contract Ministry of Economy and Finance (BES-2013-
censis para la evaluación de la calidad del agua del Río Bobos, México
064041).
Alexis Joseph Rodríguez-Romero1; Ameyalli Rios-Vazquez1; Axel Eduardo Rico-Sanchez1; Eugenia
López-López1; Jacinto Elias Sedeño-Diaz2; Myriam Catalá-Rodríguez3
Grupo de Biología y Toxicología Ambiental, Facultad de Ciencias, UNED, 28040, Madrid, España;
Universidad Nacional de Educación a Distancia.
3 Laboratorio de Ecotoxicología, Departamento de Medio Ambiente, INIA, 28040, Madrid, España.
*Presenting author contact: maquilino@ccia.uned.es
1 2
Los ríos son ambientes muy dinámicos que sostienen un flujo elevado de
materia y energía, sin embargo, están sujetos a perturbaciones por efecto de la
contaminación. La evaluación de la calidad del agua, puede ser analizada haciendo
uso de biomarcadores que midan la alteración de procesos bioquímicos en especies
centinela, ante diferentes grados de perturbación ambiental. El río Bobos se localiza
en la vertiente del Golfo de México y a lo largo de su cauce atraviesa un mosaico
con diferentes usos del suelo. Se evaluó la respuesta en la actividad mitocondrial
en esporas de Cyathea costaricensis expuestas a 11 muestras de agua a lo largo del
río, durante tres eventos de monitoreo (febrero, junio y septiembre de 2014) y se
correlacionó dicha respuesta con los usos del suelo y las variables fisicoquímicas
medidas en campo y determinadas en laboratorio. Los resultados caracterizaron
tres zonas del río, siendo la parte alta, una zona con naturaleza geológica de roca
caliza y vegetación natural de tipo bosque mesófilo de montaña, con aguas alcalinas
y concentraciones altas de sulfatos y carbonatos, esta zona mostró en junio y
septiembre los valores más altos del % de inhibición en la actividad mitocondrial
(<20%); la parte media dominada por usos de suelo derivados de prácticas agrícolas,
y aguas con mayor cantidad de nutrientes (nitrógeno y fosforo), presentó efectos
de estimulación de la actividad mitocondrial (hormesis) en los meses de junio y
septiembre y por último la parte baja asociada a usos de suelo de zonas urbanas
que se caracterizó por concentraciones altas de coliformes, cloruros, nitrógeno
amoniacal, sólidos suspendidos y temperaturas más altas, mostrando efectos de
hormesis de menor intensidad que en la parte media. La actividad mitocondrial
253
en C. costaricensi mostró ser un biomarcador sensible a las variaciones espaciales y
Evaluation of the toxic effects of phthalates on natural populations
temporales de la calidad del agua.
of Chironomus riparius (Diptera): implications for ecotoxicity studies.
O. Herrero1,2, R. Planelló1, P. Gómez-Sande3,4, M. Aquilino1, G. Morcillo1
Instituto Politécnico Nacional-Escuela Nacional de Ciencias Biológicas;
Instituto Politécnico Nacional-Coordinación Politécnica para la Sustentabilidad;
3
Universidad del Rey Juan Carlos
1
2
Chinomids,sentinel organisms for the evaluation of water quality, are considered
an outstanding model in Ecotoxicology for studying the effects of environmental
pollutants. In contrast to its widespread use in studies with laboratory cultures, the
use of natural populations (chronically exposed to complex mixtures of pollutants
in their environment) is still uncommon to assess the usefulness of molecular
biomarkers in studying the health of populations under multiple stress conditions.
Our results show that both DEHP and BBP cause rapid alterations in the activity
of GST enzyme as well as in the expression profile of various genes related to cell
stress response (hsp70 and hsc70), to the hormonal route (EcR and ERR) and to
detoxification mechanisms (CYP4G), as well as. Both compounds cause different
responses to these targets, especially in longer exposures, and have the ability to
produce delayed toxicity. The alteration of the ecdysone hormone pathway in
our experiments has special relevance, since it proves that both compounds are
endocrine disruptors in insects. Finally, this study shows differences with previous
data obtained with laboratory cultures in both the toxicity of these phthalates and
the behavior of some targets, which emphasizes the need of carrying out studies with
different populations to get a more realistic approach to the effects of contaminants.
This work was supported by the Plan Nacional de Investigación Científica, Desarrollo
e Innovación Tecnológica (Spain), grant CTM2012-37547 from the Ciencias y
Tecnologías Medioambientales program.
255
Grupo de Biología y Toxicología Ambiental, Facultad de Ciencias, UNED, 28040, Madrid, España.;
Universidad Nacional de Educación a Distancia.
3
Departamento de Zoología y Antropología Física, Universidad de Santiago de Compostela, Campus Sur
s/n, 15782 Santiago de Compostela, España., 4
Estación de Hidrobioloxía “Encoro do Con”, Universidad de Santiago de Compostela, Castroagudín s/n,
36617 Vilagarcía de Arousa, España.
*Presenting author contact: maquilino@ccia.uned.es
1 2
Evaluation of ecotoxicity of two solvents: glycerol derived solvent vs
ionic liquid
Eduardo Perales1; Cristina B. García1; Laura Lomba1; Esther Sarasa1; José I. García Laureiro2; Beatriz Giner1
3F03F (1,3-bis(2,2,2-trifluoroethoxy)propan-2-ol) is a new solvent obtained
from biomass that could replace BMIM PF6 (1-Butyl-3-methylimidazolium
hexafluorophosphate) in industrial processes. Because of its origin it is supposed
that a solvent obtained from biomass could be less harmful for the environment than
an ionic liquid.
Ecotoxicity of both solvents has been evaluated using two different
bioindicators: Vibrio fischeri (bacteria) and Daphnia magna (crustacean). The
inhibition of bioluminescence test of Vibrio fischeri was used to check the EC50 that
produces a decrease in in bacterial light productionafter 30 minutes of exposure of
bacteria to each solvent. The acute inmobilization test of Daphnia magna, following
the guidelines of the OECD 202, was employed to estimate the EC50 that produced
the lack of mobility of crustaceans after their exposure to the chemicals for 24 hours.
Although in Vibrio fischeri EC50 for both chemicals were very similar and none
of them could be considered as toxic for the environment, results in Daphnia
magna showed that BMIM PF6 is more toxic than 3F03F in this species.
Based on the differences in evaluation of ecotoxicity found in both bioindicators,
evaluation of ecotoxicity of the solvents should be checked with more biomodels of
different trophic levels.
Grupo de Investigación GreenLife. Facultad de Ciencias de la Salud, Universidad San Jorge, Villanueva de
Gállego, Zaragoza, Spain; 2
Instituto de Síntesis Química y Catálisis Homogénea (ISQCH). CSIC-Universidad de Zaragoza, Zaragoza,
Spain
1
257
Seasonal variation of micronuclei frequency in common sole of the
basque coast (SE Bay of Biscay)
Nagore Cuevas1; Izaskun Zorita1; Javier Franco1
Micronuclei (MN) and nuclear buds (NB) have been widely used as biomarkers
of genotoxicity in environmental biomonitoring studies. In order to distinguish
between the natural variability and the potential adverse effects of anthropogenic
pollution, it is essential to identify baseline levels and to establish the influence of
abiotic (season and temperature) and biotic (gametogenesis) factors. Thus, common
soles (Solea solea) were monthly collected from June 2012 to May 2013 from a
presumably pristine area located in the Basque coast (SE Bay of Biscay). Additionally,
common soles were also collected every autumn (2011-2013) from a polluted estuary.
Blood samples were withdrawn and frequencies of MN and NBs were determined in
erythrocytes. Results indicated that levels of MN and NBs recorded in the pristine area
varied among seasons, showing higher frequencies in winter and summer probably
linked to spawning and temperature increase, respectively. Accordingly, common
soles from the polluted estuary presented higher MN and NBs frequencies than
common soles from the pristine area indicating signs of genotoxic impact. However,
this genotoxicity decreased from 2011 to 2013 evidencing a lower environmental
impact. In conclusion, these findings provide vital baseline information for future
environmental biomonitoring studies using common sole as sentinel organism.
Acknowledgment: Work funded by research project CTM2012-40203-C02-02
(MINECO), Bilbao Bizkaia Water Consortium and Ministry of Agriculture, Fisheries and
Food Policy of the Basque Government.
1AZTI
259
Zinc, copper and cadmium accumulation in liver, kidney and muscle tissues
of lithobates catesbeianus tadpoles
Cleoni dos Santos Carvalho1; Tatiane Pasquoto2; Heidi S.M. Utsunomiya2; Monica Jones Costa1;
Marisa Narciso Fernandes3
The assessment of the biological responses of aquatic vertebrate species
is frequently employed to monitor water pollution, as it provides significant
information on bioavailability and actual concentration levels of several pollutants.
The concentration of metals (Zn, Cu and Cd, 1 µg L-1) on the liver, kidney and muscle
of Lithobates catesbeianus was analyzed after for 2 (acute) and 16 (chronic) days
exposure. As a result, it was observed that metal accumulation occurred in all tissues
following both acute and chronic exposures when compared to control. However,
Zn and Cu in the muscle and Cd in kidney were the highest metal accumulation
rate (2 and 16 days). The concentration of Cu and Cd in all groups associated to
Zn increased. In L. catesbeianus the accumulation of metals was in the sequence
kidney>muscle>liver. The liver is the xenobiotic detoxification organ; the muscle
and kidney were more susceptible to metals, the muscle due to high metabolic
rates (metamorphosis) and the kidney due excretion processes. The highest metal
accumulation rate was in the kidneys. Exposure to metals can generate a long-term
dysfunction in L. catesbeianus during in the larval phase; our results suggest that
further studies should be made in water ecosystems, using tadpoles as an indicator
to monitor pollutant. Financial support: FAPESP Proc. 507523/2011.
1
Universidade Federal de Sao Carlos (UFSCar), campus de Sorocaba; 2Programa de Pós Graduação em
Biotecnologia e Monitoramento Ambiental, UFSCar; 3Universidade Federal de Sao Carlos (UFSCar)
261
Transcriptional level of genes involved in the neurotransmitter system of Dicentrarchrus labrax in response to chronic exposure to psychopharmaceuticals
C.Costa1,2; R.Urbatzka3, M.Ferreira2
During the last decades, a variety of psychopharmaceuticals (PP) were
recognized as an important source of environmental contamination and
increasing concern emerged regarding their potential ecological impact. These
pharmaceuticals targets are highly conserved among vertebrates suggesting
that aquatic animals like fish are likely to be affected by environmental
exposure even at low concentration. Previous studiesof our group showed
that inDicentrarchrus labrax (D. labrax) the ratio of RNA/DNA increased after
1 day of exposure to Fluoxetine at the concentration of 0.5 and 50 µg/L.
Peroxidase enzyme activity in the plasma decreased after 1 day at the same
PP and concentration. The aim of this study was to evaluate transcriptional
levels of genes involved in D. labrax central nervous system in response to
waterborne exposure to two PP. Selected antidepressants were: Fluoxetine
(0.5µg/L and 50µg/L) and Venlafaxine (0.01µg/L and 1µg/L). The chronic
exposures were performed in a flow-through system for 21 days, with a 7 day
recovery period to assess the reversibility of effects. The brain of D. labrax was
collected and preserved in RNALater. Quantitative real-time PCR (qPCR) assays
were successfully established for genes encoding for proteins involved in the
neurotransmitter system, such as 5-Hydroxytryptamine subtype 3A (5-HT3A)
and 3B (5-HT3B), serotonin transporter receptor (SERT), monoamine oxidase
(MAO), vesicular monoamine transporter (VMAT) and dopamine 2 (D2) and
3 (D3). Specific primer pairs were designed based on UCSC Genome Browser
of the sea bass genome (http://seabass.mpipz.mpg.de/cgi-bin/hgGateway).
263
Currently, the qPCR analyses are ongoing and first results will be presented
Atrazine increases mitochondrial glutathione peroxidase activity of
about transcriptional changes of neurotransmitter genes in D. labrax. This
gills of sea lamprey juveniles during salt acclimation
study aims to establish a potential link between environmental exposure of PP,
M Candeias1; I Alves-Pereira2; MJ Lança3; AF Ferreira4; BR Quintella5; PR Almeida6; R Ferreira2
and physiological and neurological changes in fish through the expression of
the genes of their neurotransmitters.
Acknowledgements
This study was supported by FCT through the project PTDC/AAG-MAA/2405/2012 and Pest-C/MAR/
LA0015/2013.
Sea lamprey (Petromyzon marinus, L) Portuguese populations have shown a
progressive decline during the last decades. During metamorphosis and trophic
migration, gills and liver of sea lamprey juveniles undergo marked changes that
may be disturbed by oxidative stress possibly triggered by anthropogenic sources
Ciimar
1
ICBAS/UP – Institute of Biomedical Sciences Abel Salazar, University of Porto, Largo Professor Abel Salazar,
2, 099 – 003 Porto, Portugal
2
METOX – CIIMAR/CIMAR, Interdisciplinary Centre for Marine and Environmental Research, Laboratory of
the Coastal and Marine Environmental Toxicology Research Group- University of Porto, Portugal
3
LEGE – CIIMAR/CIMAR, Center Interdisciplinary Marine and Environmental Research, Laboratory of
Ecotoxicology, Genomics and Evolution - University of Porto, Portugal
1
of xenobiotic in freshwater habitats that can disturb its seawater acclimatization.
The aim of this study was to evaluate the influence of atrazine on stress markers of
gills and liver mitochondria of juvenile sea lamprey from Lima river basin during salt
acclimation. Sea lampreys sampled at the beginning of the downstream migration,
after completing the metamorphosis, were transported alive to the laboratory.
Two groups of 40 specimens were hold in tanks with 50 or 100 μg/L atrazine,
during 30 days, and the salinity was gradually increased to 35, following a three
step procedure. Control groups were treated identically except without atrazine
exposure. Mitochondria obtained by centrifugation of tissues homogenates in 50 mM
Tris-HCl pH 7.5 buffer were used to determinate reactive oxygen species, glutathione
and glutathione disulphide contents, and superoxide dismutase and glutathione
peroxide activities. The exposure of sea lamprey juveniles to 50 μg/L atrazine, in the
salt gradient conditions, caused an increase in the level of mitochondrial non-protein
thiols and reactive oxygen species in gills, while the exposure to 100 μg/L atrazine
caused an increase of glutathione peroxidase activity which was correlated with a
decrease in the reactive oxygen species content. In contrast, significant changes in
these mitochondrial stress markers in liver and in the superoxide dismutase activity
and glutathione/glutathione disulphide ratio of mitochondria in both tissues were
265
not detected.The efficient response ensured by glutathione peroxidase, against
Microplastics in fish species of three Portuguese estuaries
oxidative stress in sea lamprey juveniles, during salt acclimation, points out this
N. Castro1,2, Ferreira, M.3, Urbatzka, R.4, Ramos, S.2
enzyme as a promising marker of exposure to elevated levels of this triazine herbicide.
Microplastics (<5 mm) are widespread in marine and fresh water ecosystems.
Concerns exist about potential physiological damage to a wide range of organisms
ICAAM - Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da
Mitra, Ap. 94, 7006-554 Évora, Portugal;
2
ICAAM - Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Departamento de Química,
Universidade de Évora, Ap. 94, 7006-554 Évora, Portugal;
3
ICAAM - Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Departamento de Zootecnia,
Universidade de Évora, Núcleo da Mitra, Ap. 94, 7006-554 Évora, Portugal;
4
MARE, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal;
5
MARE, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal and
Departamento de Biologia Animal, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1746016 Lisboa, Portugal;
6
MARE, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal e
Departamento de Biologia, Escola de Ciências e Tecnologia, Universidade de Évora, Largo dos Colegiais
2, 7002-554 Évora, Portugal
1
and about bioaccumulation of toxic pollutants adsorbed on plastic. Unlike the
marine environment, presence and effect of microplastics in freshwater ecosystems
remain relatively unknown. This work aimed to study the ingestion of microplastics
by fish fauna of the Douro, Lima and Minho estuaries. Several species were studied,
including macrobenthic feeders (Platichthys flesus, Dicentrarchus labrax), detritivorous
(Chelon labrosus, Liza ramada) and omnivorous (Diplodus sargus). A total of 145
individuals were collected, including 21 adults and 124 juveniles, by beam-trawl
and pushing-nets. Microplastics ingestion was analyzed through visual observation
of stomach contents, made under a stereoscope microscope. Prior to the opening,
all of the stomachs were rinsed with filtered distillated water, as well as all of the
instruments, to prevent external contamination. A control glass petri dish was placed
near by the working area, to check for airborne contamination, which was negligible.
Considering all stomachs, 29 (20%) contained microplastics, and the most abundant
form were fibers. Adult fishes (33%) had a higher frequency of microplastic ingestion
than juveniles (18%). Douro was the most impacted estuary, with an ingestion rate
of 22%, compared with Lima (16%) and Minho (9%). Macrobenthic feeders were the
most contaminated species (38%), above detritivorous (14%) and omnivorous (0%),
pointing out to species-specific differences in terms of microplastic ingestion. This
study includes the first analysis of contamination by microplastics in Portuguese
estuarine fish fauna and results are discussed in the scope of environmental risk
related with these emergent contaminants.
267
Acknowledgements
This study was supported by FCT through the project Pest-C/MAR/LA0015/2013.
An application of fish gill histopathology to address the interaction
ICBAS/UP – Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Largo Professor Abel
Salazar, 2, 099 – 003 Porto, Portugal
2 Ecobiotec, CIIMAR/CIMAR – Centro Interdisciplinar de Investigação Marinha e Ambiental, Laboratório de
Toxicologia Ambiental – Universidade do Porto, Rua dos Bragas n.º 289, 4050-123 Porto, Portugal
3
METOX, CIIMAR/CIMAR – Centro Interdisciplinar de Investigação Marinha e Ambiental, Laboratório de
Toxicologia Ambiental – Universidade do Porto, Rua dos Bragas n.º 289, 4050-123 Porto, Portugal
4 LEGE, CIIMAR/CIMAR – Centro Interdisciplinar de Investigação Marinha e Ambiental, Laboratório de
Toxicologia Ambiental – Universidade do Porto, Rua dos Bragas n.º 289, 4050-123 Porto, Portugal
Marta Martins1; Maria Helena Costa1; Pedro M. Costa1
1 effects between different polycyclic aromatic hydrocarbons (PAHs)
Gills are the main apical entry organ of waterborne toxicants in fish, whether
dissolved or bound to suspended particles. Although not the principal organ
involved in xenobiotic biotransformation (unlike liver), gills are in direct contact
with contaminated water and may endure significant lesions, compromising
critical physiological functions, from gas exchange and osmoregulation to
excretion. Aquatic sediments tend to trap contaminants, especially more
hydrophobic, like PAHs, which may be released to the water column, e.g.
following disturbance. In spite of the importance of gills to the health of fish,
establishing clear cause-effect relationships between sediment-bound toxicants
(isolated or mixed) and histopathology needs enhancement. The present study
addressed the interactions between two model PAHs, benzo[b]fluoranthene
(B[b]F) and phenanthrene (Phe), considered to be carcinogenic and noncarcinogenic to fish, respectively, taking the seabass Dicentrarchus labrax, as
an ecologically-relevant model. For the purpose, 28-day laboratorial bioassays
with spiked sediments were conducted (with individual and combined PAHs
at equitoxic and realistic concentrations). Semi-quantitative and quantitative
histopathological were enforced in the gills, based on weighted indices and
epithelial cell measurements, respectively. Overall, PAHs induced significant
alterations, either isolated or combined, despite their low, ecologicallyrelevant, concentrations. Epithelial lifting and interlamellar hyperplasia were
the most noticeable alterations, together with evidence for inflammation.
Individuals exposed to B[b]F presented higher severity and diffusion of lesions,
269
especially at day 28. Changes to chloride cells also occurred. Interestingly, the
Assessing the seasonal variability of biotic integrity indices in a mediterra-
mixture that caused the most significant effects comprised the highest and
nean stream exposed to industrial sewage discharge: a multi-taxa approach
lowest concentrations of B[b]F and Phe, respectively, whereas the remaining
Nicole Colin1; Alberto Maceda-Veiga1; Nuria Flor-Arnau12; Josep Mora3; Pablo Fortuño-Estrada3;
Cristiana Costa-Vieira4; Narcis Prat3; Jaume Cambra2; Adolfo de Sostoa1
treatments failed to reach the level of alterations recorded for exposure to
isolated B[b]F, revealing potential antagonistic effects. These may be caused
by impairment of metabolic responses to exposure and show that fish gill
histopathology, when addressing mixtures of toxicants, may yield unexpected
results that can compromise assessment for the purpose of biomonitoring.
Studies on the response of aquatic biota to sewage discharges along an
upstream-downstream gradient are central to assess the recovery ability of rivers
and the diagnostic accuracy of monitoring tools. Using indices of biotic integrity, this
study examined the effects of an industrial sewage discharge on the community of
macrophytes, diatoms, macro-invertebrates and fish along 1.5 km in a Mediterranean
MARE - Marine and Environmental Sciences Centre
1
stream. Relationships between water quality variables (ammonium, nitrite, nitrate,
oxygen concentrations, pH, temperature, and conductivity), flow and five indices of
biotic integrity (IPS, IMF, IBMWP, IMMi-T and IBICAT) were determined and compared
between impacted sites (P1, P2 and P3) in relation to a reference site (R1) across
seasons in 2012. In all seasons, the discharge site of the effluent from the industrial
sewage treatment plant (P1) scored lower in all indices of biotic integrity than
other polluted and reference sites. Along the downstream pollution gradient, the
community integrity of macrophytes, diatoms and macro-invertebrates progressively
improved but only that of diatoms (IPS) and macro-invertebrates (IBMWP) scored
similar to R1 in summer. In spite of biological communities should mirrored changes
in the water quality variables, attributed to increased or decreased dilution ability,
some assemblages are influenced by augment of flow. Diatoms (IPS) are strongly
correlated with flow, therefore is not a suitable indicator in seasonal assessment,
because the pollution effect is masked. Fish occurrence was restricted to the sites
R1 and P3 throughout the study, however in the last station IBICAT values is lower.
According with results, IBMWP (macroinvertebrates) and IMF (macrophytes) indices
could be the most suitable because the recovery pattern is less influence by natural
271
constraints and work more adequately to pollution influence. In summary, multi-taxa
Effects of crude extract of cyanobacteria (Radiocystis fernandoi) on the kidney
approach across seasons permits to determine taxa and indices more suitable for
of traíra (hoplias malabaricus): oxidative damage and histophatology
pollution effect assessments to temporal scale in stream systems.
Marisa N Fernandes1; Marise M Sakuragui1; Edneide M Brasil1; Naiara E S Souza1; Priscila R Siqueira1; Marcelo G Paulino2; Driele Tavares1; Ana P. Terezan1; João B. Fernandes1; Alessandra Giani3
Department of Animal Biology, University of Barcelona (UB), Spain;
Department of Botany, University of Barcelona (UB), Spain;
3
Department of Ecology, University of Barcelona (UB), Spain;
4
InBIO - Rede de Investigação em Biodiversidade e Biologia Evolutiva, Laboratorio Associado, Spain
1
2
The microcystins (MC) are toxins produced by cyanobacteria that may increase
the free radicals and induce oxidative stress and histopathological changes in
different organs of fish. The kidney is one of the first organs to be affected by
MC as it participates of excretion route. In this context, this study evaluated the
toxicity of MCs present in the crude extract of Radiocystis fernandoi on the kidney
of traíra, Hoplias malabaricus. Fish were separated in groups and received an
intraperitonial injection (i.p.) every 72 h during 30 days, control group (i.p. saline
0.9%, SA30D) and MC group (i.p. 100 μg kg-1 body mass, MC30D). Thereafter,
fish were killed andthe kidney were sampled for morphological analyses and
determine the activity of ethoxyresorufin O-deethylase (EROD), superoxide
dismutase (SOD), catalase (CAT), glutathione-s-transferase (GST), glutathione
peroxidase (GPx) as well as the levels of glutathione (GSH) and lipid peroxidation
(LPO). EROD activity and GSH levels decreased and the activities of SOD, CAT, GPx
and the levels of LPO increased. The more frequent histological alterations were
nuclear and cellular hypertrophy, cellular vacuolization, melanomacrophages
aggregates, granular degeneration, occlusion of the tubular lumen and focal
tubular necrosis of epithelial cells, with loss of nuclei, fragmentation of cell.
This alteration is very severe stage III and the damage caused to the tissue is
irreversible coagulative necrosis. The results suggest that after 30 days of MCs
injection induced SOD, CAT and GPx in kidney. The increased levels of LPO indicate
oxidative stress in the MC group. Furthermore, the lesions in the kidney were
severe and classified in the stage III. LPO and histopathology is a good biomarker
273
of chronic MCs contamination. Financial support: CAPES Proc. 2276/2011, CNPq/
Effects of temperature and emerging contaminants of high concern on
INCT-TA Proc. 573949/2008-5 and CEMIG Proc. GT346.
juveniles of the common goby (Pomatoschistus microps)
Elsa Fonte1; Pedro Ferreira1; Lúcia Guilhermino1
Universidade Federal de São Carlos;
2Universidade Fedral de São Carlos; 3Universidade Federal de Minas Gerais
1
The contamination of the environment by emerging compounds of high concern
such as pharmaceuticals and microplastics (MP) is priority research topic at international
level. More knowledge on the effects of these environmental contaminants is needed
to improve the basis of ecological and human risk assessments, especially those
induced by simultaneous exposure to different substances (mixtures) and in relation to
alterations resulting from global climate changes (e.g. temperature increase). Thus, the
objective of the present study was to investigate the effects of the antibiotic cephalexin,
alone and in mixture with MP, on juveniles of the common goby (Pomatoschistus
microps) at two distinct temperatures (20º C and 25ºC). Early juveniles of P. microps were
collected in the Minho River estuary and acclimated to lab conditions. The bioassays
were carried out at the two temperatures, by exposing juveniles individually to
different concentrations of cephalexin in the presence and absence of MP for 96h.
After the exposure period, the predatory performance and sub individual biomarkers
were determined. The results indicated that exposure to cephalexin (low ppm range)
induces toxic effects on fish, and that temperature rise and the presence of MP may
influence the cephalexin-induced toxicity. Therefore, more studies on mixture toxicity
under temperature variation scenarios are needed to adapt and mitigate the predicted
effects of global climate changes.
ICBAS & CIIMAR, University of Porto; ICBAS, Depart Populations Study, Lab of Ecotoxicology,R. Jorge
Viterbo Ferreira, 228, 4050-123 Porto, Portugal; CIIMAR, Research Group of Ecotoxicology, Stress Ecology
and Environmental Health, R dos Bragas, 289, 4050 Porto
1
275
Effect of adsorbed TiO2 nano-particles on aquatic organisms.
Marta Sendra Vega1; Ignacio Moreno Garrido1; Luis María Lubián1; Antonio Tovar Sánchez1; Julián
Blasco Moreno1
Titanium dioxide nanoparticles (TiO2 NPs) are widely used in multitude of applications
and present a potential risk due to the release to the marine and freshwater systems.
TiO2 NPs have been demonstrated to be toxic to unicellular algae, as they can be adsorbed
or internalized by the cells. TiO2 NPs could induce toxicity to aquatic organisms through
various mechanisms including physical damage, shading effect, oxidative stress and/
or internalization. Surface interactions, such as adsorption of NPs oncells, are important
toxicity mechanisms. One of the deleterious could be the increase of cell weight due to the
adsorption of NPs, which would sink the floating cells out from the photic zone. On the other
hand, different NPs can present different surface properties and aggregation states in the
NP-cell suspensions, leading to different NP-cell heteroagglomerations. Water chemistry,
such as pH and ionic strength (IS), can also influence the aggregation state of NPs and will
thus influence the NPs-Cell.Since sizes of both microorganism cells and NPs aggregates are
within the scale of colloids, the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory can be
applied to analyze the colloidal behavior of NPs and cells in water.
In this study, different freshwater and seawater species of microalgae were chosen in
order to investigate interactions between algal cells and TiO2 NPs and TiO2bulk, as well
assedimentation rate of NPs, exposed and non exposed,cells in the water column. This is
the first study specifically designed in order to check the NP-cell heteroagglomeration and
cells sedimentation in water systems. Our findings show new light on bio-nano interaction.
1
ICMAN-CSIC, Instituto de Ciencias Marinas de Andalucía CSIC, Campus Río San Pedro, 11510 Puerto Real
(Cádiz) Spain.
E-mail contact: marta.sendra@icman.csic.es
277
Mercury bioaccumulation in piscivorous fishes of an Amazon reservoir,
Brazil
Sarah Sampaio Py-Daniel1; Daniele Kasper2; Bruce Rider Forsberg2; Eurizângela Pereira Dary2;
Jansen Alfredo Sampaio Zuanon2; Efrem Jorge Gondim Ferreira2; Wanderley Rodrigues Bastos3;
Olaf Malm4
Mercury has molecular forms more susceptible to absorption by aquatic
fauna than metallic mercury. Mercury levels were used as a trophic level
determinant when studying fish, although accumulation rates of this element
can be altered by construction of reservoirs. Here we determinate total
mercury levels (THg) in muscle tissue of same trophic level (piscivorous fish)
in the Balbina’s reservoir, Brazilian Amazon. Additionally we investigated
the influence of standard length, species and habit of life in mercury levels
of piscivorous fish. Fish samples (n=148; 8 taxa) were collected between
September and October 2011. THg were determined on dorsal muscle tissue
by acid digestion. The detection and quantification of THg were performed
in a CVAAS (FIMS-400, Perkin–Elmer). Average mercury levels of piscivorous
fish was 281 ± 290µg/kg wet weight, and differed significantly between
species (KW=37.751; p<0.0001). THg levels of five taxa (Cichla spp., Agoniates
halecinus, Plagioscion squamosissimus, Serrasalmus rhombeus, Pygopristis
denticulata) were positively correlated with standard length of each. Only
three taxa (Acestrorhynchus spp., Ageneiosus spp., Cynodon septenarius)
showed no correlation between THg levels and standard length. Piscivorous
fish with benthopelagic habit showed highest THg levels (U’=3412;
p<0,05), except for Pygopristis denticulata. Among the four pelagic species
(Acestrorhynchus spp., Ageneiosus spp., Cynodon septenarius, Cichlaspp.),
three showed values below THg level maximum recommended limit for
279
human consumption (500 µg/kg wet weight) established by World Health
Use of the brain european sea bass (Dicentrarchus labrax) cell line
Organization (WHO). But among the four benthopelagic fish species(Agoniates
DLB-1 for toxicological studies
halecinus, Plagioscion squamosissimus, Serrasalmus rhombeus, Pygopristis
Patricia Morcillo1; Jose Meseguer1; María Ángeles Esteban1; Alberto Cuesta1
denticulata), two showed results above the WHO limit. We show that piscivorous
fish have variations in mercury levels in muscle tissue relative to standard
Nowadays, the contamination of aquatic habitats with heavy metals from various
length, species and life habit. Therefore, differences in mercury levels among
industrial and mining sources is a serious environmental problem not only for the
species of piscivorous fish in the reservoir were greater than among feeding
aquatic communities but for consumers. In this sense, fish are known to be the
habit.
greatest inputs of toxic elements for humans. Therefore, evaluation of the toxicity of
aquatic pollutants in fish and especially in marine fish cell lines as an alternative to fish
bioassays is needed in order to throw some light in the heavy metal toxicity. In this
Keywords
study, a new cell line, derived from the European sea bass brain (DLB-1), obtained in
trophic level, total mercury levels, carnivores guild
our laboratory, was exposed for 24 h to cadmium (Cd), methylmercury (MeHg), lead
Universidade de Brasília;
2
Instituto Nacional de Pesquisas da Amazônia;
3
Universidade Federal de Rondônia;
4
Universidade Federal do Rio de Janeiro
1
(Pb) and arsenic (As) being the cytotoxicity examined. The four heavy metals produced
a dose-dependent cytotoxicity as determined by neutral red (NR) and MTT-tetrazolio
(MTT) assays. Cd showed the highest toxicity for DLB-1cells (NREC50=0.004mM;
MTTEC50=0.0010mM) followed by MeHg, As and Pb. Furthermore, exposure of DLB1 cells to the EC0 and EC50 doses of the heavy metals induced cellular protection by
the up-regulation in the expression of the methallothionein-A gene (mta), mainly by
As.This work reports the use of a new marine fish cell line (DLB-1) for toxicological
purposes and concretely to evaluate the potential of heavy metals on marine waters
and fish.
University of Murcia
1
281
Enantiospecific chronic effects of ibuprofen in the freshwater crustacean,
A. desmarestii.
Elena Nieto1; Miriam Hampel2; Jaume Aceña3; Enrique Gonzalez-Ortegon1; Sandra Perez3; Pilar
Drake1; Julian Blasco1
The development of advanced analytical techniques have allowed to detect the
occurrence and fate of pharmaceutical compounds in different matrixes such as ground
water and sediment, either in their parent compound or in the form of metabolites. Many
of these compounds have at least one asymmetrically substituted carbon atom and are,
therefore chiral. There are very few studies about the chiral behavior of chemicals in the
aquatic environment and the potential negative effects in the organisms. An example
of these compounds is Ibuprofen (IB); a widely used nonsteriodal anti-inflammatory
drug (NSAIDs). IB is formed by two enantiomer forms: R and S. In order to improve the
knowledge about the different ecotoxicological effects that different enantiomeric forms
can have in aquatic organisms, we performed a series of toxicity tests on the freshwater
shrimp Atyaephyra desmarestii as model organism. A. desmarestii is widely distributed in
clean freshwater bodies and has been successfully employed in pharmaceuticals ecotoxicity
tests. Selected sublethal endpoints were osmoregulatory capacity and ingestion- and
respiration rate as indicators of physiological changes. The shrimps were exposed at IB in its
composite form and both enantiomeric forms at environmental relevant concentrations.
Given that different enentiomers of a chiral compound can cause different biological
effects or the presence of one enantiomer may inhibit the metabolism of the other, it is
important know not only the fate but also the effect of these substances in non-target
species to improve the environmental risk assessment of chiral compounds.
Institute for Marine Science of Andalusia, Department of Ecology and Coastal Management;
Andalusian Center for Marine Science and Technology, CACYTMAR;
3
Institute of Environmental Assessment and Water Research, Department Environmental Chemistry
1
2
283
Effects of mercury bioaccumulation in gills of wild fish (liza aurata)
assessed by 1h nmr metabolomics and oxidative stress endpoints
F Brandão1; T Cappello2; O Albuquerque1; S Guilherme1; MA Santos1; M Maisano2; A Mauceri2; M
Pacheco1; P Pereira1
Oxidative stress has been described as a key pathway to initiate mercury (Hg)
toxicity in fish. However, Hg toxicity mechanisms in fish still need to be clarified. To this
aim, the application of environmental metabolomics, in combination with a battery
of oxidative stress biomarkers, appears as an innovative and effective approach
that can provide important contributions for the elucidation of such mechanisms.
Hence, this comprehensive strategy was applied to the gills of golden grey mullet
(Liza aurata) inhabiting an Hg contaminated system (Aveiro lagoon, Portugal). The
higher accumulation of inorganic Hg and methylmercury in gills of fish from the
contaminated area (Laranjo - LAR) was followed by marked changes both in metabolic
profile and antioxidant responses, pointing out a compromised health status. Nuclear
magnetic resonance (NMR)-based metabolomics displayed a clear differentiation
between contaminated (LAR) and uncontaminated (São Jacinto - SJ) areas, revealing
changes in metabolites related to antioxidant protection. In detail, depletion of
reduced glutathione (GSH) and its constituent amino acids, glutamate and glycine,
was observed in fish from LAR. The interference of Hg with the antioxidant protection
of gills was corroborated through oxidative stress endpoints, namely the depletion
of glutathione peroxidase and superoxide dismutase activities at LAR. Moreover, the
increase of total glutathione content (reduced glutathione+oxidized glutathione) at
LAR, in parallel with the GSH depletion above mentioned, indicates the occurrence of
massive GSH oxidation under Hg stress, and an inability to carry out its regeneration
(glutathione reductase activity was unaltered) or de novo synthesis. Nevertheless,
the results suggest the occurrence of alternative mechanisms for preventing lipid
285
peroxidative damage. Overall, the novel concurrent use of metabolomics and
Fish response to the elevated potassium level: the model of the mining
oxidative stress endpoints demonstrated to be sensitive and effective towards a
area contamination.
mechanistically based assessment of Hg toxicity in gills of fish.
E.V. Borvinskaya1; I.V. Sukhovskaya1; O.B. Vasilyeva1; L.P. Smirnov1; S.A. Svetov2; N.A. Krutskih2
Mining sites and related facilities for the preparation and processing of ores are known
Department of Biology and CESAM, University of Aveiro, 3810-193 Aveiro, Portugal;
2
Department of Biological and Environmental Sciences, University of Messina, 98166 Messina, Italy
1
potential environmental polluters. In the North-West of Russia there is a lake receiving
processing wastes from local iron mine and ore dressing mill. During the mill operation
the lake with ultra fresh water was transformed to the reservoir with low-mineralized
water, withmarked potassium and sulfates accumulation. Potassium present at an
anomalous concentration (to 172 mg*L-1) compared to other macroelements (sodium
to 20 mg*L-1, calcium to 19 mg*L-1and magnesium to 37 mg*L-1), which results in the
K-SO4-HCO3 type of water instead of the regional Ca - HCO3 type. The dramatic decline
in biodiversity, which was observed in the lake in a rage of plankton communities to
fish, suggests a possible toxic effect of excessive level of potassium on the fresh water
organisms. The present study was aimed to test whether the shifted ratio of K+ and
Na+ cations the contaminating factor of emerging concern. Whitefish Coregonus
lavaretus was subjected to experimental conditions with water with a ratio potassium
and sodium close to that in the contaminated lake. The results demonstrate strong
inhibitory effect of water with excessive potassium on the fish growth. The level of
products of lipid peroxidation also changed significantly, indicating the oxidative stress
in fish tissues. Glutathione S-transferase activity and reduced glutathione content
demonstrated low sensibility of components of xenobiotic biotransformation system
to the studied factor. These data are of interest for the potential risk assessments of
mining and related industrial activities on the regional water ecosystems.
Institute of Biology of Karelian Research Centre Russian Academy of Sciences, Russia;
Institute of Geology of Karelian Research Centre Russian Academy of Sciences, Russia
1
2
287
Annual cycle of the microzooplanktonic aloricate and loricate ciliates abundances in a eutrophic lagoon system of the central Gulf of California, México
Aída Martínez López1*; A. Pérez-Morales2; G. A. Ayala-Rodríguez2
Microzooplankton includes small crustacean nauplii, copepods, rotifers,
heterotrophic ciliates, foraminifera and radiolarian. The heterotrophic ciliates, mainly
composed of aloricate ciliates, are the main component of the microzooplankton
community. In general, microzooplankton plays an important role in subtropical
coastal systems because they consume up to 70% of phytoplankton biomass
(pico- and nanoplankton). These phytoplanktonic components are the numerically
dominant size fractions in the central region of the Gulf of California, México.
Nonetheless their importance, basic information of microzooplankton and their
potential top-down control has been overlooked in this region. This study is the first
monthly compilation, and will serve as a baseline for future surveys that document
abundance and seasonal variability of two microzooplankton components (aloricate
ciliates and tintinnids) at the eutrophic complex lagoon of Topolobampo-Santa
María-Ohuira. Total abundance ranged from 2 to 56 x 103 ind./L and showed their
highest numbers in winter-early summer. The minimum numbers were recorded in
August-September. The aloricate ciliates were the dominant component throughout
the year at all sampling stations. Although higher mean values were reached in
Ohuira lagoon, the maximum abundance for the data series was recorded at Santa
María lagoon, when the phytoplankton bloom occurs. The density distribution
revealed that the substantial increases in the abundance of the microzooplankton
components quantified were related to wastewater discharges at sites with
exacerbated eutrophication conditions.
289
Key words
Microzooplankton, eutrophic lagoons, annual cycle, Mexico.
Evidencia de efectos subletales en daphnia magna expuesta a sucesivos
shock tóxicos de licor negro
Centro Interdisciplinario de Ciencias Marinas, Instituto Politécnico Nacional;
Instituto de Ciencias Marinas y Pesquerías, Universidad Veracruzana
1
Departamento de Plancton y Ecología Marina, Centro Interdisciplinario de Ciencias Marinas (CICIMARIPN), A.P. 592, C.P. 23096, La Paz, Baja California Sur, México;
2
Instituto de Ciencias Marinas y Pesquerías, Universidad Veracruzana. Calle Hidalgo No. 617, Col. Río
Jamapa, C.P. 94290. Boca del Río, Veracruz, México.
*e-mail: diatomeas@yahoo.com.mx
1
Soledad Chamorro Rodríguez1
2
Evidencias de derrames accidentales de licor negro en plantas de celulosa,
pueden
ingresar al sistema de tratamiento de efluentes. Considerando que
las características de este residuo líquido son distintas al influente normal, su
ingreso al sistema de tratamiento y posterior disposición sobre los ecosistemas
receptores, puede tener repercusiones negativas para el medio ambiente. En este
sentido, ensayos enfocados en respuesta subletales sobre Daphnia magna, están
considerándose como factor de estudio para evaluar inicialmente una perturbación
ambiental. El objetivo de este estudio fue determinar la toxicidad letal (LC50) y
subletal (deformidad en el organismo), a través de D. magna expuesta a shock de
licor negro. Para ello, se consideró 4 concentraciones de licor negro 2, 4,10 y 30 mL
licor/L efluente. Se realizaron ensayos de toxicidad aguda y subletal con D. magna
(20 individuos) expuestas a 5 concentraciones (100, 50, 25, 12,5 y 6,25%) para cada
shock. Los resultados evidencian que el efluente no presenta toxicidad aguda en
ninguno de los shocks tóxicos, sin embargo, presenta alteraciones principalmente
a nivel de rostrum y espina caudal. De este modo, el shock 1 presenta alteración
a la concentración de 50 y 100%, evidenciando un 10% de deformidad en la
espina caudal para ambas concentraciones y un 20% sobre el rostrum al 50%. Por
su parte el shock 2, presenta un 20% de deformidad en espina caudal a 25 y 50%.
Shock 3 solo evidencia alteraciones al 25% con 20% en espina caudal y 30% en el
rostrum. Por último, shock 4 presenta alteraciones en todas las concentraciones
(6,25 - 100%), con un 30% en la espina caudal y 50% en el rostrum al 50%.
291
Agradecimientos FONDECYT N° 3120216,
1120664 Y CONICYT/FONDAP/15130015.
1Grupo de Ingeniería y Biotecnología Ambiental, Facultad de Ciencias Ambientales y Centro EULA-Chile.
Universidad de Concepción. Chile
Soil Pollution, Conservation And Restoration
PAHs in soils affected by forest fires in Serra do Caramulo (Central Portugal)
Joana Paula Machado Ribeiro1; Pedro Dias1; Marcos Bastos1; Cândida Garcia1; Catarina Mansilha2; Jorge Espinha Marques1; Deolinda Flores1
Serra do Caramulo is a mountain located in the Central Portugal, with a maximum
elevation of 1077 m and an area of about 500 km2. Serra do Caramulo is located in the
Central-Iberian Zone of the Iberian Massif. The dominant geological units are PrecambrianCambrian metasedimentary rocks and Variscan granitic rocks. In the summer of 2013,
Serra do Caramulo was affected by three intense forest fires, resulting in a joint burnt
area of approximately9415.5 ha. As a consequence, the environmental impacts are evident
and therefore measures for recovery, rehabilitation and protection should be carried out.
For that purpose, the comprehensive identification of the effects on the environment and
the characterization of the affected materials are essential. The aim of the present study
is to identify the effects of the referred forest fires in soils from Serra de Caramulo, where
samples from affected and non-affected areas were collected. In addition, samples were
collected in areas dominated by both granitic and metasedimentary lithology’s, in which
soils present different physic-chemical characteristics and consequently are differently
affected by the forest fires. Considering that polycyclic aromatic hydrocarbons (PAHs)
are products of in­complete combustion processes, the samples were analyzed by gas
chromatography coupled to mass spectrometry for the identification and quantification
of the 16 priority PAHs. The results demonstrate that, generally, the concentration of some
priority PAHs, especially high molecular weight PAHs, increases in soil samples collected
in areas from metasedimentary lithology’s and affected by the forest fires. Some PAHs
ratios are generally used for source identification, as is the case of Flt/(Flt+Pyr) which was
selected to evaluate the effect of biomass burning. The results obtained for soils from
293
Serra do Caramulo, i.e., Flt/(Flt+Pyr) > 0.5 in samples from areas affected by the forest fires
How does metal(loid) pollution affect soil enzymatic activities at
identifies biomass combustion as a source of PAHs.
metal(loid) enriched mine tailings? The role of pioneer plants rhizospheres
Departamento de Geociências, Ambiente e Ordenamento do Território, Faculdade de Ciências da Universidade do Porto;
Instituto Nacional de Saúde Doutor Ricardo Jorge
1
Héctor M. Conesa1; María-Nazaret González-Alcaraz2; Francisco-José Jiménez-Cárceles3; María del
Carmen Tercero-Gómez1; Isabel Párraga-Aguado1
2
The phytomanagement by phytostabilisation of metal(loid) enriched tailings
has been proposed as a feasible technique to decrease their environmental
risks. Plants may immobilise metal(loid)s within their rhizospheres preventing
leaching, decreasing erosion, enhancing soil microbiology and thus promoting
shifts in ecological successional processes. Taking advantage of pioneer
vegetation which spontaneously colonises tailings may be a good option to
restore tailings, above all in semiarid areas, where plants must also cope with
drought and high evapotranspiration. The goal of this work was to assess the
selection of spontaneous plant species for the further phytomanagmenent of
mine tailings attending to the amelioration properties of their rhizospheres.
For this purpose, a transect-designed sampling from non-disturbed soils
(control) to mine tailings was performed, and the description of edaphic
gradients (including soil microbiology assessed by soil enzymatic activities)
was performed. The extent of the improvement achieved in the rizhospheres
of spontaneous vegetation was also shown. The main differences when
comparing the tailings with the control site were mainly determined by salinity
(and the corresponding water extractable ion concentrations), soil fertility,
microbial biomass and activity and metal(lolid) concentrations (total and water
extractable). High dense spontaneous vegetation patches inside tailings called
“fertility islands” showed similar edaphic properties as peripheral areas. The
enhancement of soil microbiology in plant rhizospheres was species-dependent:
grasses<shrubs<<trees≤fertility islands. Tree species seem to be the key for the
295
establishment of “fertility islands”. Small rises in the soil microbiology within
Chemical characteristics and toxicity of soils from an abandoned Pb/Zn mine
tree rhizospheres may trigger important shifts in plant successional processes.
Manoel Lago-Vila1; Daniel Arenas-Lago1; Andrés Rodríguez-Seijo1; María Luisa Andrade1; Flora
Alonso Vega1
Funding: MINECO of Spain and FEDER (CTM2011-23958) and Fundación Séneca
of Murcia (15296/PI/10). Dr. Héctor M. Conesa thanks MINECO and UPCT for
Ramon y Cajal programme (RYC-2010-05665). Dr. María-Nazaret GonzálezAlcaraz holds a post-doctoral grant from Fundación Ramón Areces.
This work is aimed at assessing the total content and extractable forms of Cd,
Pb and Zn in the soils developed in a Pb/Zn abandoned mine (Galicia, NW Spain).
The toxicity of the soils for Brassica juncea (L.) Czern., Sinapis alba L.,Festuca ovina L.
and Cytisus scoparius (L.) Link was evaluated. Three different areas were selected;
Departamento de Ciencia y Tecnología Agraria, Universidad Politécnica de Cartagena, Cartagena, Spain;
Department of Ecological Science, Faculty of Earth and Life Sciences, VU University, Amsterdam, Netherlands;
3
TECOMA Ingeniería y Ambiente S.L., Murcia, Spain
1
2
two of them are minespoils (R1 and R3) and one is the settling pond (R2). Three soils
in each one were sampled and exhaustively characterized by determining their
chemical properties and the extractable and total contents of heavy metal. Soil
pollution was evaluated using the pollution index (IP). Biotoxicitytestwas performed
to determine chronic toxicity of the soil samples on higher plants. Parameters
calculated were the percent inhibition of seed germination (IG), percent root length
inhibition (IR), and percent germination index (GI). Toxicity classification system was
used for assessing the soil toxicity. Results show that total Cd concentration varies
between 28-44, Zn between 21,374-32,287, and Pb between 1,737 and 6,761 mg kg1. IP values indicated that all soils are severely polluted by Pb, Zn, and Cd. Germination
index were 26-82% for S. alba, 62-253% for B. juncea, 99-128% for F. ovina, and
19-43% for C. scoparius. These values indicate strong inhibition and thus toxicity
of these soils for S. alba and C. scoparius, and less effect, even stimulation, for B.
juncea and F. ovina (phytoremediator species). In general was observed an increase
in metal toxicity measured by the biotoxicity test with increasing extractable metal
contents in soils. The negative correlation between CaCl2 and LMWOA extractable
content of Cd, Pb and Zn and GI, and positive with IR indicate that available content
affect adversely the germination index and root elongation. The biotoxicity tests are
a suitable complement to evaluation of soil pollution.
297
Acknowledgements
This study was supported by the Ministerio de Economía y Competetividad (Project CGL2013-45494-R)
and Xunta de Galicia (EM2013/018).
Effects of multiple environmental stressors (thermal stress and cadmium
exposure) at different times and levels of biological complexity in E. fetida
Nerea Garcia-Velasco1; E. Urionabarrenetxea2; M. Gandariasbeitia2; A. Irizar2; Manu Soto1
Universidade de Vigo
Departamento de Biología Vegetal y Ciencia del Suelo. Lagoas. Marcosende. 36310, Vigo. Spain
email: manolago@uvigo.es
1
Terrestrial ecosystems are subjected to multiple environmental stressors due
to the climate change derived from global warming and to the increase in the
synthesis, manufacturing and release of chemicals. According to IPCC predictions
the most feasible effects to occur in soils are related to temperature raises, moisture
changes, acidification and hypoxia phenomena. Hence is crucial to assess how
a multi stressor scenario can affect the responses of terrestrial organisms when
assessing soil health. Eisenia fetida earthworms have been used in standard toxicity
tests (OECD, ISO) and a variety of biomarkers have been developed in them at
different levels of biological complexity (population, organism, cellular) in order
to assess soil health. Recently, the Neutral Red Uptake (NRU) in vitro assay with
coelomocytes from exposed earthworms has been optimized as an efficient tool for
toxicity assessment of metallic compounds and real soils (complex mixtures). The
aim of this work was to study the effects produced in predicted scenarios defined by
the co-occurrence of thermal and chemical (Cd) stresses at different times and levels
of biological organization with the aid of the earthworm reproduction test (OECD222) and NRU assay. Adult earthworms were maintained in LUFA soils (standard soil
no. 2.3) at optimal temperature (19°C) and subjected to thermal stress (26°C) in soils
with CdCl2 (0-125 mg Cd/kg) for 3 and 14 d. Mortality, weight loss and soil and tissue
Cd concentrations were quantified, complementarily NRU assay was performed in
extruded coelomocytes. Effects on reproduction (number of cocoons, juveniles)
were determined after 56 d. Reproductive impairment, accumulation of Cd, weight
loss and cytotoxic effects on coelomocytes were enhanced after exposure to Cd and
299
thermal stress at longer exposure periods. The obtained results prove the usefulness
Copper fractionation in high resolution sampled vineyard soils man-
of the earthworm reproduction test (OECD-222) and NRU to forecast the effects of
aged under heroic viticulture in Ribeira Sacra (NW Spain)
multi stress scenarios in soil health.
P Pérez-Rodríguez1; P Otero-Pérez1; D Fernández-Calviño2; J C Nóvoa-Muñoz1; M Arias-Estévez1; E
Álvarez-Rodríguez3; A Núñez-Delgado3; M J Fernández-Sanjurjo3
Acknowledgements
Funded by the Basque Government (Grant to Consolidated Research Groups; IT810-13), University of the
Basque Country (UFI 11/37)
1PIE-UPV/EHU;2UPV/EHU
Long-term application of Cu-based fungicides leads to Cu accumulation
in the uppermost layers of vineyard soils. However, some soil properties and
management practices could modify this general trend which is neglected if soil
sample collection comprises wide depth intervals (0-20 or 0-30 cm). In this study
are shown the detailed vertical patterns (samples of few cm of thickness) of total
Cu (CuT) and its fractionation in the first half meter of three terraced vineyard
soils (PM, PT and AM) from D.O. Ribeira Sacra (Galicia, NW Spain).
The highest CuT values occur in the uppermost layer (0-2 cm) of PM and PT
soils (643 and 892 mg kg-1, respectively), with a sharply diminution in the 2-4 cm
layer (368-379 mg kg-1). In the AM soil sample of 0-2 cm, CuT content was 193 mg
kg-1 keeping rather uniform up to 30 cm deep (152-197 mg kg-1). However, PM
and PT soils show a steadily decrease of CuT with the lowest values (67-71 mg kg1
) in the deepest soil sample (40-50 cm).
Organically-bound Cu and residual Cu (that belong to the crystalline
structure of minerals) are the main Cu fractions comprising on average 43 and
30% of CuT, respectively. Copper associated to inorganic non-crystalline Al and
Fe oxyhydroxides (CuIA) and to crystalline Al and Fe oxyhydroxides (CuC) are
quite similar being about 12% of CuT, although CuIA is somewhat higher in the
uppermost soil layers (0-2, 2-4 and 4-6 cm) and CuC in the deeper ones. Average
of exchangeable Cu fraction is 2.4% of CuT, being slightly higher (up to 8%) in
deeper layers of AM soil.
The detailed vertical pattern of CuT and its fractionation must be interpreted
301
in terms of potential Cu mobilization in vineyard soils and consequent risk of Cu
Total Copper in the bulk and rhizosphere soil of wild plants grown in long-
appearance in surrounding surface waters.
term contaminated vineyard soils from d.O. Ribeira Sacra (NW Spain)
P Pérez-Rodríguez1; E Pena-Calvo1; J C Nóvoa-Muñoz1; M Arias-Estévez1; M J Fernández-Sanjurjo2;
A Núñez-Delgado2; E Álvarez-Rodríguez2
Universidade de Vigo;
University of Copenhagen;
3
Universidade de Santiago de Compostela
1
2
Despite high total Cu content in long-term managed vineyard soils, often with
values of soil total Cu (CuT ) above 200 mg kg-1, some spontaneous plants are able
to grow suggesting that their root system can manage the Cu bioavailability,
mainly in the rhizosphere.
In this study, the CuT of bulk soil samples (<2 mm, 0-20 cm depth) of three
vineyard soils from D.O. Ribeira Sacra at NW of the Iberian Peninsula (PM, AM
and OU) are compared to CuTdetermined in loosely adhering rhizosphere (hand
shaken detached root soil, LAR) and in tightly adhering rhizosphere (mechanically
shaken detached root soil, TAR), obtained from the most frequent wild plants that
occur in the selected vineyards: Digitaria sanguinalis (G1), Setaria viridis (G2), Picris
hieracoides (C1), Conyza albida (C2), Tolpis barbata (C3),Chenopodium
album (R1), Phytolaca americana (SB) and Rumex induratus (R2). Fifteen samples
of LAR and sixteen samples of TAR where analyzed besides to three bulk soil
samples.
Total Cu in the vineyard soils ranged from 190 mg kg-1 in AM to 353 mg kg-1 in
PM, whereas LAR samples showed 1.4-2.5 times more CuT than bulk soil with an
average of 479 mg kg-1(range 262-930 mg kg-1). LAR samples from G2 and R1
plants collected in PM vineyard are those with the highest CuT values (>800 mg
kg-1). For TAR samples, CuT ranges from 105 to 1004 mg kg-1 (average 470 mg kg-1),
being C1 the plant with the highest CuT value (1004 mg kg-1).
Total Cu enrichment in rhizosphere samples (LAR and TAR) claims a new study
on Cu fractionation in these samples due to CuT is a poor indicative of metal
303
mobility. This will help to assess if the rhizosphere environment can modify Cu
Microbial loop structure and occurrence of heavy metal resistant bacteria
bioavailability and mobility compared to bulk soil.
in sediments of Araça Bay (Brazil): influence of contaminants
Ana Julia Fernandes Cardoso de Oliveira1; Bruna Del Busso Zampieri2; Vanessa da Costa Andrade2;
Antonia Cecilia Zacagnini Amaral3
Universidade de Vigo;
Universidade de Santiago de Compostela
1
2
This study aimed to analyze the structure of the microbial loop and the
occurrence of resistant bacteria to heavy metals in sediments of the Bay of Araca,
São Sebastião (SP), Brazil, due to organic contamination caused mainly by the Port
of San Sebastian by the Pipelines and Terminals South Centre (DTCS). Sediment
samples were collected in 37 points, divided into two regions (intertidal and
subtidal), during the period of one year. The samples were analyzed for particle
size distribution, organic matter content, total organic carbon, hydrocarbons,
and microbial components. The highest densities of microorganisms were
obtained from the subtidal samples. There was a negative correlation between
the densities of heterotrophic bacteria and nanoflagellates, which may indicate
bacterivory and the variations of organic matter directly affected populations
of heterotrophic bacteria. The strains of heterotrophic bacteria isolated during
the study were evaluated for their resistance to heavy metals. Bacteria strains
tested were more resistant to zinc (Zn) (56%) and less resistant to copper (Cu)
(10%). After Zn, Chromium (Cr) was the heavy metal for which the bacteria had
higher percentages of resistance (54%) which is especially important because
Cr is used as anticorrosive in paints for ships. The Cr is used in industry and craft
as anticorrosive substances and petroleum refining. Only 23% of the strains
were not resistant to heavy metal, and 49% were resistant to more than two
simultaneously heavy metals and 7% of the bacteria tested were resistant to
co-Cadmium (Cd, Cu), Cr and Zn, thus, multi-sturdy. When compared to the
resistance of bacteria metals tested in two sampling regions was observed that
305
the bacteria isolated from sediment subtidal region were more resistant to Zn,
Movilización de cobre por salpicadura de la lluvia en suelos de cultivo de vid
Cr and Cu than those isolated from intertidal region. Only for cd most resistant
P Pérez-Rodríguez1; D Soto-Gómez1; M Paradelo1; J E López-Periago1
bacteria was present in the intertidal. Responses of benthic flora, as obtained
in this work are useful information to analyze the impact of pollution by heavy
El enriquecimiento en cobre de la superficie de suelos dedicados a viñedo por
metals and other contaminants and are of great importance to public health
tratamientos de base cúprica presenta un riesgo de movilización de partículas con
due to bacterial co-resistance to metals and antibiotics.
elevados contenidos de cobre. Un proceso relevante que inicia la movilización de
suelo es la desagregación y suspensión por impacto de la lluvia. El objetivo fue
obtener evidencias de la movilización de Cu por salpicadura de la lluvia en suelos
UNESP - Campus do Litoral Paulista;
2
UNESP - Instituto de Biociências de Rio Claro; 3UNICAMP Instituto de Biologia
1
con una historia de aplicación de cobre.
Se estudiaron dos parcelas monitorizadas con colectores de salpicadura situados
entre de hileras de vid conducidas en espaldera (Ribadavia y Castrelo do Miño,
Ourense España), con control de adventicias por erradicación entre pasillos y una
historia de tratamientos foliares de base cúprica de más de 10 años.
Se aplicó una dosis de 2,1 y 1,2 kg Cu ha-1, en 2009 y 2010, respectivamente. Se
determinaron muestras de salpicadura lateral en bandejas de captación (8 cm de
altura de borde y 162 cm de perímetro), volumen de salpicadura, masa de sólidos y
concentración en las fracciones CuS (< 0,45 micras), y CuP (> 0,45 micras), tras cada
episodio de lluvia registrado (pluviometro 0,2 mm/pulso). Las proyecciones laterales
de salpicadura se expresaron en masa por metro lineal de superficie horizontal.
Los resultados muestran valores medios de sólidos traslocados de 0,18 ± 0,20 g
m-1. La correlación entre la cantidad de sólidos y la intensidad máxima diaria es débil.
La correlación con parámetros de energía erosiva de la lluvia es no significativa.
El pH medio de la suspensión de la salpicadura fue 6,24 ± 0,26. La riqueza en cobre
307
de los sólidos proyectados (0,8 ± 0,88 g kg-1) produjeron una movilización de 0,16 ±
Particle size characterization of throughfall from vine leaves sprayed with
0,21 mg CuP m . La concentración de CuS 0,026 ± 0,39 mg L supera valores tóxicos
Cu-based fungicides, by DLS, AF4-MALS and SP-ICP-MS
legales (>0,022 mg Cu L , DOCE, 2006). Todas las muestras con pH<6,2 superaron
P Pérez-Rodríguez1; D Soto-Gómez1; M Paradelo1; J E López-Periago1; I De La Calle1
-1
-1
-1
este umbral. La relación de concentraciones [CuP]/[CuS] son del orden de 104 L kg-1,
mucho mayor que las relaciones de reparto de adsorción.
Cu-based fungicides applied on vine leaves to fight against fungal
diseases in crops are lost in washed-off by rainfall. Cu particle size analysis
Referencias
Arias et al., 2007 Influence of aging on copper fractionation in an acid soil.
European regulations for the quality of continental waters (law 2006/44/CE of DOCE, 2006) (Diario official
de las Comunidades Europeas, 2006).
Universidade de Vigo
1
by different methods was done on throughfall samples in a monitored
vineyard
sprayed
with
copper
oxychloride
(particle
size
0.979
µm). The objectives of this work were: to evaluate the presence of Cu in micro and
nanoparticles in throughfall after several rainfall episodes. Two throughfall samples
were analyzed by: dynamic light scattering (DLS) which was used to evaluate
the presence of particles; asymmetric-field flow-field fractionation multiangle
laser light scattering (AF4-MALLS) was used to calculate the hydrodynamic
diameter of the particles and size fractionation. Finally, single-particle mode
inductively coupled plasma mass spectrometry (SP-ICP-MS) allows measuring the
presence of Cu in suspended nanoparticles. Particle sizes are shown in Table 1. Results showed that there are no significant differences among the three techniques.
Colloids released in throughfall contain Cu and have a size smaller than they had when
they were applied. These results suggest Cu from treatments dissolves and combines
with natural colloids (e.g., airborne deposition or organic matter from the leaf surface)
and then released by rainfall. Deeper studies of Cu-nanocompounds should be done
due to their size may result in a high potential mobility in agroecosystems.
Table 1. Results obtained from two samples of throughfall by different techniques
309
DLS
Sample
Episode 1
Hydrodynamic
diameter (nm)
135.9
Episode 3
120.5
AF4-MALLS
SP-ICP-MS
Hydrodynamic Mean size of particles
diameter (nm)
containing Cu (nm)
130.6
140.1
113.4
Study of the competitive release of heavy metals (Cu, Zn, Ni, Cd and Pb)
in a copper mine soil treated with ground mussel shell
L. Cutillas-Barreiro1; L. López-Armada1; J.C. Nóvoa-Muñoz1; M. Arias-Estévez1; A. Núñez-Delgado2;
M. J. Fernández-Sanjurjo2; E. Álvarez-Rodríguez2
118.7
A stirred flow chamber experiment was carried out to assess the effect of ground musell
shell (GMS) addition in heavy metal release (Cu, Zn, Ni, Cd and Pb) by a mine soil. A control
Universidade de Vigo
1
soil sample (C), from the upper 20 cm of a dump of an Cu mine (NW Spain), and this soil
amended with GMS at two rates (12 and 48 Mg ha-1) were spiked with a solution containing
1.57 mmol L-1 of Cu, Zn, Ni, Cd and Pb (NaNO3 0,01M as background electrolyte). Control
mine soil (C) and amended mine soil (C-GMS12 and C-GMS48) were incubated in the
dark at field capacity during 24 h and 1 month, being the loss of water checked at daily
basis and compensated by adding distilled water. After the incubation periods, 0,2 g of
sample (C, C-GMS12 and C-GMS48) was placed in a stirred flow chamber reactor made of
polypropylene with a chamber volume of 1,5 cm3. A NaNO3 0.01M solution was circulated
at a flow rate of 0,6 mL min-1, collecting 80 samples in vials (one each 3,33 min). The
concentration of heavy metals in the extracts was determined by AAS.
In the extracts from control mine soil, the heavy metal release sequence was
Cu>Zn≥Ni>Cd>Pb. A remarkable diminution of the concentration, close to the limit of
detection, for all the heavy metals studied was observed in the mine soil amended with the
highest dose of GMS (48 Mg ha-1), especially in the samples incubated during one month.
The strong acidity of the studied mine soil promotes heavy metal mobility, so the
addition of GMS is showed as an efficient strategy for its immobilization as well as it can be
considered as an alternative for its reutilization.
Department of Plant Biology and Soil Sciences, Faculty of Sciences, University of Vigo, 32004 Ourense, Spain;
Department of Soil Sciences and Agricultural Chemistry, Higher Polytechnic School, University of
Santiago de Compostela, 27002 Lugo, Spain
1
2
311
Mercury distribution among soil aggregate size fractions in spodic
horizons of temperate forest podzols from Galicia (NW Spain)
L. Bibián-Núñez1; L. Cutillas-Barreiro1; A. Gómez-Armesto1; X. Pontevedra-Pombal2; M. Arias-Estévez1; E. García-Rodeja Gayoso2; J. C. Nóvoa-Muñoz1
Mercury is an atmospheric global pollutant which is mainly accumulated
in the uppermost soil layers. However, Hg can be mobilized downwards as
it happens in podzolic soils where illuvial horizons (Bh, Bhs, Bs) are the last
barrier that avoid its arrival to groundwaters and surface waters. Thus, it could
be important to understand what is the Hg fate in these horizons could be
closely related to grain size fractions, as they content distinctive amount of
compounds (organic C, Al and Fe oxyhydroxides) involved in Hg immobilization.
This study asses the distribution of Hg in aggregate size fractions (coarse sand,
fine sand, coarse silt, fine silt and clay) of eight spodic horizons from temperate
forest podzols. The aggregate size fractions were separated by wet sieving
(sand fractions) and centrifuging using Stokes´ law for finer fractions. In each
fraction, total Hg (HgT) was determined using a Hg analyzer (Nippon MA-2000).
In the bulk soil (< 2mm), HgT values range from 11 to 292 ng g-1 being
exceeded the critical load threshold in soils (130 ng g-1) in three of the
eight horizons analyzed. Regarding aggregate size fractions, the highest
concentrations of HgT were obtained for the clay fraction (125-2075 ng g-1)
whereas in sand fractions (fine and coarse) HgT vary between 0,7 and 60 ng
g-1. Surprisingly, coarse sand showed higher HgT concentrations (0,7-254 ng
g-1) than fine sand (1,7-87 ng g-1). Coarse and fine silt showed intermediate
values, being somewhat higher for fine silt (38-836 ng g-1) than for coarse silt
(7-228 ng g-1).
From these results, it is expected that a distinctive content of organic C and
313
Al and Fe oxyhydroxides in the aggregate size fractions can respond to the
Distribution and accumulation of total hg in high vertical resolution
observed Hg distribution.
sampled temperate forest podzols from Galicia (NW Spain)
Department of Plant Biology and Soil Sciences, Faculty of Sciences, University of Vigo, 32004 Ourense, Spain;
Department of Soil Sciences and Agricultural Chemistry, Faculty of Biology, University of Santiago de
Compostela, 15872 Santiago de Compostela, Spain
1
A. Gómez-Armesto1; C. Ferro-Vázquez2; L. Cutillas-Barreiro1; M. Costas-Casais3; M. Arias-Estévez1;
J.C. Nóvoa-Muñoz1; A. Martínez-Cortizas4
2
Despite the significance of the atmosphere in the global Hg cycle, soils are the
main Hg reservoir in terrestrial ecosystems where Hg is mostly accumulated in
the uppermost layers due to its affinity to bind to soil organic matter. However,
sometimes greater concentrations of Hg are found in subsurface horizons than
in surface soil layers, as it occurs in podzols.
This study describes preliminary results on Hg in two podzols (ACB-I and
ACB-II) from Galicia (NW Spain), focused in the vertical distribution of Hg
concentration and accumulation (as areal mass basis) through a high vertical
resolution sampling strategy (5 cm intervals). Total Hg (HgT ) in the A horizon
varies between 12-49 and 19-31 ng g-1 for ACB-I and ACB-II, respectively,
showing a decreasing trend with depth. In both soils, the lowest concentrations (HgT < 10 ng g-1) were found for the E horizons, whereas samples corresponding
to illuvial horizons (Bhs) showed the largest concentrations throughout the soil
(HgT 12-73 ng g -1 for ACB-I and 17-74 ng g -1 for ACB-II). Total Hg in Bw horizons
shows a decreasing trend with depth (from 39 to 21 ng g-1 in ACB-I and from 59
to 37 ng g-1 in ACB-II).
Regarding the Hg reservoir, both soils accumulate a significant proportion of
the element in the illuvial horizons (Bhs: 19.7 and 18.9 mg Hg m-2 in ACB-I and
ACB-II soil respectively). On the other hand, E horizons scarcely accumulated
Hg and their reservoir is below 2.6 mg Hg m-2.
These results suggest that Hg is mobilized from superficial to deeper horizons,
becoming an issue that should be studied in detail due to the risk of Hg release
315
to groundwaters, in particular under climate scenarios (i.e. warming) that may
Contenido de mercurio en suelos y sedimentos de cuencas hídricas afect-
promote changes in pedogenetic routes of non-climacic soils (regression of the
adas por la actividad antrópica (Ushuaia, Tierra del Fuego, Argentina)
podzolisation processes).
S. Diodato1; A. Moretto2; J. Escobar1; J. Marcovecchio3; L. Cutillas-Barreiro4; A. Gómez-Armesto4; M.
Arias-estévez4; J. C. Nóvoa-Muñoz4
Departamento de Biología Vegetal y Ciencias del Suelo, Facultad de Ciencias, Universidad de Vigo, 32004
Ourense, España;
2
Instituto de Ciencias del Patrimonio-CSIC, Edificio Monte da Condesa bajo, 15872 Santiago de Compostela,
España;
3
Departamento de Geografía, Facultad de Geografía e Historia, Universidad de Santiago de Compostela,
15872 Santiago de Compostela, España;
4
Departamento de Edafología y Química Agrícola, Facultad de Biología, Universidad de Santiago de
Compostela, 15872 Santiago de Compostela, España
1
El mercurio (Hg) es un elemento de elevada toxicidad ambiental y gran
habilidad para ingresar en los sistemas biológicos, que alcanza los suelos
mediante deposición seca, húmeda y la caída de biomasa senescente. Ushuaia
(Sur de Tierra del Fuego, Argentina), es una ciudad rodeada por bosques
caducifolios de Nothofagus pumilio por donde discurren cursos de agua que
desembocan en la zona costera atravesando la zona urbana. En este estudio se
seleccionaron tres cuencas cuya parte final atravesaba al entorno urbano, y una
cuarta (como control) en el Parque Nacional Tierra del Fuego. En cada cuenca
se ubicaron un lugar en la zona alta, en la zona media y en la desembocadura
como representación de áreas de nula, media y elevada influencia urbana. En
los 12 lugares se tomaron muestras superficiales de suelos y sedimentos en
primavera de 2013, y en verano y otoño de 2014, en los que se determinó el
contenido y la distribución de Hg y otros parámetros químicos. Los niveles de
Hg en suelos variaron entre 21 y 77 ng g-1, mientras que en sedimentos variaron
entre 23 y 128 ng g -1. El Hg en el suelo se correlacionó positivamente con el
N, S, P y algunos metales (Cr, Zn, K, y Ca), mientras que en los sedimentos las
correlaciones fueron positivas con C, N, P, S, Fe, Zn, Na, K y Mg, y negativas con
el pH. Un análisis multivariante mostró que los suelos de la desembocadura
del Arroyo Grande (compuesto por material de relleno) y los sedimentos de
la desembocadura del Arroyo Buena Esperanza en la Bahía Encerrada (con
aporte continuo de efluentes pluviocloacales sin tratar) presentaron los niveles
más altos de Hg, S, P y cationes. Esto resultados sugieren que las actividades
317
antrópicas urbanas alteran el estado natural de los ecosistemas.
Evaluation of riparian vegetation and invasive species in Galician river
ecosystems
X. Álvarez1; F., Abilleira1; E., Valero1
Centro Austral de Investigaciones Científicas (CADIC- CONICET), B. Houssay 200, Ushuaia, Argentina;
Universidad Nacional de Tierra del Fuego (UNTDF), Onas 450, Ushuaia, Argentina;
3
Instituto Argentino de Oceanografía (IADO- CONICET-UNS), Camino La Carrindanga km 7,5, Bahía Blanca,
Argentina;
4
Departamento de Biología Vegetal y Ciencias del Suelo, Facultad de Ciencias, Universidad de Vigo, 32004
Ourense, España
1
2
The biological diversity are affected by the destruction and degradation of
natural habitats. Specifically, pressure from human activity on a large number of
the riparian zones in Pontevedra has led to the modification and loss of habitats,
which then affect biotic communities and may lead to changes in species
richness, species composition and species relative abundance. This phenomenon
occurs especially in stream banks and in the adjacent lands (considering its
characteristics suitable for agriculture and livestock). And it has reduced what
could be a gallery forest into a line of trees or has even eliminated it altogether in
some areas. This means that the protection the woods provide to the soil, among
many other functions, is lost. For this reason it is necessary to restore the riparian
woodland from a conservation point of view, without forgetting that it is also
a point of reference for the ecological, environmental and scenic quality of the
area. To achieve this goal, the river ecosystem was evaluated with the QBR index,
to characterize the quality of riparian forest, and the RQI was also proposed for
assessing the structure and functioning of riparian zones based on hydrological
and geomorphological conditions. Finally, all invasive species which were found
in the banks were characterized. The condition of the riparian forest was heavily
linked to land use, and consequently the ecological quality of the river. In urban
areas the conditions were higher, but also in agricultural and pastoral ones where
riparian forest has been removed. The spring of the river were found in better
condition, with minor anthropogenic influences. The application of these indexes
is relevant if the requirements of WFD are taken into account.
319
Universidad de Vigo
1
AF4 Research Group. Engineering Department of Natural Resources and Environment, Forestry
Engineering College, University of Vigo, Campus A Xunqueira s/n., 36005 Pontevedra, Spain.
email: xaalvarez@uvigo.es. Tel.: +34-986-801-903; Fax: +34-986-801-907.
1
Contamination of waters, stream sediments, soils and plants from the
abandoned Murçós W-Sn mine area (NE Portugal)
Martins,L.1; Antunes, I.m.2; Gomes, E.p.1; Neiva, A.m.r.3; Teixeira, R.j.1; Neiva,A.m.r4
The mining complex of Murçós belongs to the Terras de Cavaleiro Geopark,
located in Trás-os-Montes region, northeast Portugal. A stockwork of NW-SEtrending W-Sn quartz veins intruded Silurian metamorphic rocks and a Variscan
biotite granite. The quartz veins contain quartz, muscovite, tourmaline, cassiterite,
wolframite, scheelite, arsenopyrite, pyrrhotite, pyrite, sphalerite, chalcopyrite,
galena, native bismuth, stannite and also later bismuthinite, joseite, matildite,
anglesite, zavaritskite and roosveltite. The exploitation of W and Sn produced 335
t of a concentrate with 70 % of scheelite and 150 t of another concentrate with
70 % of cassiterite between 1948 and 1976. The exploitation took place mainly
in four mine open pits, but also underground. Three open pits, each with a lake,
were left in the area. Remediation processes of confination and control of tailings
and rejected materials and fitoremediation with macrophytes from the lakes
were carried out between 2005 and 2007. The abandoned mining area is located
close to a rural area.Water samples were collected in 2008. Those from the open
pits are acidic (pH=4.03) and contain the highest electrical conductivity value
(EC=343 µS/cm), while surface water samples are neutral (pH=7.05 ; EC=25.5 µS/
cm). They have low metal concentrations, but are contaminated in As, Fe, Mn,
Al, Cd, Ni and SO42-, mainly in the dry season. In general, stream sediments are
not contaminated. Only two of them are moderately to heavily contaminated
in Bi and W, respectively. Soils are contaminated in As and must not be used
for agriculture. Metals and As concentrations of macrophyte fresh water from
separated roots and leaves do not show any significant bioconcentration.
321
Department of Geology, University of Trás-os-Montes e Alto Douro and CEMUC Centre, Portugal;
Polytechnic Institute of Castelo Branco and CIGAR/FEUP Centre, Portugal;
3
Department of Earth Sciences, University of Coimbra and GEOBIOTEC Centre, Portugal;
4
U COIMBRA
1
2
Exploring the Potential of the Zeolite in Contaminated Soil
Ana Katsarova1; M. Hristova1; N. Dinev1
Heavy metal pollution is a big problem for industrialized countries. Basic pollutants
of soil, plant and water are Cd, Zn, Pb, As, etc. As a result of the activities of many
industrial factories around 360,000 acres in Bulgaria soils contain Cd, Zn, As, Cu above
the maximum permissible concentration. Zeolite as a natural material is considered
to be a natural heritage in Bulgaria. A incubation experiment with soil samples from
critical areas in the country was set up in order to study the zeolite applicability in
agriculture. The soil samples have been selected for the purpose of environmental
observation near Chelopech (the gold and copper extraction plant) as a part of
monitoring network in the area. Data about soil acidity and the availability of heavy
metals have been obtained. The experiment was conducted in three different levels
of amelioration of zeolite (1%, 5%, 10%). Changes in the monitored parameters
were reported after completion of the experiment. These results indicate that visible
positive changes were obtained at a level of 5%to 10%. The aim of our investigation
was to establish effects of amendment with natural zeolite of acid and polluted soil.
Institute of Soil Science, Agrotechnologies and Plant Protection “N.Poushkarov”, Sofia, Bulgaria
1
323
Aquaculture and Environment
Microbiological contamination of Sargassum sp from north coast of São
Paulo state (Brazil) by resistant bacteria
Ana Julia Fernandes Cardoso de Oliveira1; Grazielle Stradiotto2; Wagner Vilano1; Célia Regina
Gouveia de Souza3; Aline Bartelochi Pinto2; Mirella Massonetto Basilio1
Sargassum macralgae belongs to the Phaeophyta Phylum that comprise all the
brown algae totaling about 1500 species. The brown algae dominate rocky shores
occurring from low water level to a depth of 30m. In the tropics, there are extensive
masses of floating Sargassum that is referred to as Sea of ​​Sargassum. The present
study aimed to isolate fecal indicator bacteria, Escherichia coli and Enterococcus sp,
in Sargassum algae collected in two polluted beaches at Ubatuba (Brazil), and to
evaluate the resistance of isolated bacteria to antibiotics (amoxicillin+clavulanic
acid 30µg, ampicillin 10µg, ciprofloxacin 5µg, erythromycin 15µg, streptomycin
10µg, gentamicin 10µg, rifampicin 5µg, tetracycline30µg e vancomycin 30µg).
Bacteria strains were isolated through the Membrane Filter technique with agar
mEnterococcus and modified mTEC agar media. Typical bacteria colonies were
isolated on BHI agar and resistance tests was done by Kirby Bauer Disk Difusion
method recommended by the National Comittee for Clinical Laboratory Standards.
Regarding to E. coli isolated from Enseada beach, 100% showed resistance to seven
antibiotics, except streptomycin and rifampicin, to which all strains were sensitive.
All the strains (100%) isolated from Perequê Mirim beach were resistant to seven
antibiotics and 50% of the strains were resistant to rifampicin and tetracycline.
100% of Enterococcus sp isolated from Enseada beach, showed resistance to eight
antibiotics except ciprofloxacin. All strains isolated from Perequê Mirim beach
were resistant to rifampicin, 60% to erythromycin, 20% to ciprofloxacin and 40% to
325
the other six antibiotics. The results showed that the majority of Sargassum algae
Environmental Chemistry Monitoring
analyzed were contaminated by fecal indicator bacteria and the isolates showed
resistance to several antibiotics. The high frequency of bacterial resistance is
Distributions of nutrients and metals in a sediment core and in superficial
extremely worrying since it may offer risk to human health once this algae has been
sediments from a reservoir used for public water supply São Paulo, Brazil
used in food industries and, the presence of resistant bacteria in the environment
S. Cardoso-Silva1; F. G. S. Beghelli1; M. Pompêo2; V. Moschini-Carlos1
and other organisms used for human consumption, can contribute to dissemination
of resistance genes in marine ecosystem.
Paiva Castro reservoir is one of the five reservoirs of Cantareira system, the major
drinking water source of the metropolitan region of São Paulo. The reservoir is
subjected to an intense population growth and anthropogenic uses throughout
UNESP - Campus do Litoral Paulista;
2
UNESP - Instituto de Biociências de Rio Claro;
3
Instituto Geológico SMA-SP
1
its watershed that can lead to an increment in nutrient and metal levels. A study
was conducted to investigate spatial and temporal heterogeneity of nutrients
(nitrogen and phosphorus), organic carbon (OC), sulfide and metals (Cd, Cr, Cu, Ni,
Pb, Zn, Mn, Fe) in the sediments of this important reservoir. Sampling of surface
sediments was performed at nine stations along the reservoir and in one core at
the limnical zone. In the superficial sediments were taken measurements of pH, EH
temperature and dissolved oxygen in sediment-water interface. The core was dated
using 210Pb technique. The OC content was measured indirectly by method of
ignition, phosphorus and nitrogen levels by spectrophotometry and the metals were
analyzed using atomic absorption spectroscopy. Regarding the surface sediments,
variables suggested that phosphorus was not being released for the water column
as well as the metals. It was also observed increasing levels of nitrogen (r2=0.98)
and phosphorus (r2=0.75) upstream-downstream while OC had a decrease toward
the dam (r2=0.76). The reason C:N indicated the predominance of allochthonous
organic matter in fluvial reservoir area with a gradient toward the dam indicating
predominance of autochthonous organic matter. In the core, phosphorus, nitrogen,
carbon and Cu had increasing levels over time. A principal component analysis
327
indicated that the variables nitrogen, phosphorus and Cu influenced the period from
Metal and PAH assessment of the coastline and estuaries in the north of
1987 to 2010, time of great urban expansion in the watershed. It is necessary that
Portugal
public policies are actually applied to counteract the degradation of this important
M A Salgado1;2; C M Almeida2; C Teixeira2; P Reis2, M C Basto2;3
ecosystem; otherwise, the reservoir’s ´good quality´ will be at risk.
Pollution of coastal zones has become a worldwide problem during recent
years, due to human population growth and consequent anthropogenic
Acknowledgements
FAPESP 2013/08272-0, 2012/11890-4.
activities such as industrial effluents, municipal sewage, agricultural pollutants
and accidental spills of toxic chemicals, which increased rapidly pollutants
1UNESP;
2USP
inputs into the natural environment. Metals and organic compounds such as
polycyclic aromatic hydrocarbons (PAHs) are among the major contaminants of
particular concern due to their persistence, susceptibility of bioaccumulation
and the possibility of reaching toxic levels.
Monitoring the quality of the coastal environment and estuaries is a
demand for management and conservation purposes. Estuaries in particular,
are most vulnerable to pollutants and they represent ideal habitats for
feeding, spawning and nursing for several organisms, therefore constant
surveillance is crucial.
In this study concentrations of metals and PAHs were assessed both in
water and in sediments of the northwest coastline of Portugal and in two
important estuaries (Douro and Minho) of this area.
Metals were analysed by atomic absorption spectrophotometry and PAHs
by gas chromatography with mass spectrometer detector.
Results showed that metals in coastal and in estuarine waters were mostly
below or within background levels according to Norwegian guidelines.
In estuarine sediments metals were higher than in the coastline, Douro
registering higher concentrations than Minho. Mercury was one of the metals
329
reaching levels above background, in both estuaries.
PAHs in coastal waters were undetected and in estuarine waters only
naphthalene was found. On coastal sediments naphthalene and phenantrene
The implementation of an abiotic typology for brazilian reservoirs
Sheila Cardoso-Silva1; Teresa Ferreira2; Julio Cesar López-Doval3;
Viviane Moschini-Carlos1; Marcelo Pompêo3
were present but in estuarine sediments several low molecular weight PAHs
predominated.
The typology is one of the first steps in the implementation process of the Water
Concentrations of metals and PAHs were low showing good environmental
Framework Directive (WFD), the European water resources management. The aim
conditions in selected area, but attention must be paid to estuaries to assure
of the typology is to allow that reference conditions are correctly established and
that concentrations of these chemicals are kept to a minimum.
that the classifications of the ecological status in each category of ecosystems
be similar. Study has been developed in São Paulo state in order to examine the
This work was funded by the Project ECORISK (reference NORTE-07-
possibility of adapting the WFD to the Brazilian reality. It was selected 49 São
0124FEDER-000054) within the SR&TD Integrated Program MARVALOR -
Paulo reservoirs monitored by the local environmental agency. It was applied a
Building research and innovation capacity for improved management and
typology according system A with the variables: depth, dimension and altitude
valorization of marine resources, supported by the Programa Operacional
from each reservoir. It was also applied a typology according system B with the
Regional do Norte (ON.2 – O Novo Norte) and by the European Regional
variables: depth, dimension, volume, drainage area, rainfall, altitude, longitude
Development Fund; and by the Portuguese Foundation for Science and
and latitude. A cluster analysis (Euclidian distance, Ward´s method) revealed the
Technology (FCT) through PEst-C/MAR/LA0015/2013.
formation of four kinds of reservoirs in each of the implemented systems. In system
To the ECORISK team who participated in samples collection.
A, however, reservoirs with distinct dynamics were considered similar. System B
was considered more adequate once it included specific variables for reservoirs.
Applying a principal component analysis for system B it was observed that the
1ICBAS - Instituto de Ciências Biomédicas de Abel Salazar, Rua Jorge Viterbo Ferreira, 228, 4050-313 Porto,
Portugal
2 CIIMAR - Centro Interdisciplinar de Investigação Marinha e Ambiental, Rua dos Bragas, 289, 4050-123,
Porto, Portugal
3 FCUP – Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal
variables altitude and dimension opposed large reservoirs with mean altitude
located in the west side of the state to lower reservoirs at high altitudes in the east.
Indicating, the existence of two major ecological types related to the dimension
and altitude of the reservoirs. The others subtypes were related to the depth
and longitude (secondary gradients expressed in the 2nd axis). One of the main
difficulties to apply a typology to Brazil refers to the difficulty in obtaining data. It
is suggested, for future works, the inclusion of geological data as recommended by
the WFD as well as the validation of the typology with a biological group.
331
Financial support
Fapesp, 12/11890-4, 13/08272-0, 14/22581-0.
Global amounts of polychlorinated biphenyls (PCBs) in the Porto coast-
1UNESP- Programa em Ciências Ambientais, Campus Sorocaba, Brasil;
2Instituto Superior de Agronomia Universidade Técnica de Lisboa, Portugal;
3USP, IB, Departamento de Ecologia, Brasil
MJ Rocha1,2; C Cruzeiro1,2; AS Caramelo1; S Amaral1; J Sousa2; E Rocha1,2
line and Douro River estuary
The growth of industrial zones in the Oporto district led to the emergence
of a large number of anthropogenic organic compounds that occur in several
habitats, including surface waters. In this vein, our group has produced
monitoring data sets for the presence of xenoestrogenic compounds (Rocha et
al. 2013), pesticides (Rocha et al. 2012) and polycyclic aromatic hydrocarbons
(PAHs) (Rocha et al. 2011). Nonetheless, up to this moment no information was
ever published concerning the levels of PCBs in surface waters of the Oporto
region. For this evaluation, during 2011 (n= 6), water samples taken from Leça
and Matosinhos (seacoast) and from Fluvial and Freixo (Douro River estuary) were
analysed by gas chromatography – mass spectroscopy. Results of the analysis of
39 PCBs dissolved in water (DW) or adsorbed to suspended particulate matter
(SPM), revealed that the most polluted site was the Matosinhos beach, Σ39PCBs ≈
130 ng/L (DW) and ≈ 95 ng/L (SPM), respectively. Besides, the maximal amounts of
PCB77 ≈ 5 ng/L (DW) and ≈ 11 ng/L (SPM), PCB126 ≈ 47 ng/L (DW) and ≈ 24 ng/L
(SPM), PCB189 ≈ 13 ng/L (DW) and ≈ 6.2 ng/L (SPM), considered as dioxin-like
PCBs, were surprisingly high in the area, when compared with others observed
worldwide. In this sense, since there is no information about the possible lethal
impacts of this environmental mixture of PCBs over local fauna, a toxicity test
using Artemia salina (ARC-test) will be done to start investigating this aspect.
The latter data, together with those observed during the chemical monitoring
process will provide new information about the impact of the evaluated PCBs in
this region.
333
Rocha et al. (2013). J Water Health, 11(2), 281-296.
Rocha et al. (2012). Int J Environ Anal Chem, 92(14), 1587-1608.
Rocha et al. (2011). J Chromatogr Sci, 49(9), 695-701.
Acknowledgements
PTDC/MAR/70436/2006.
Institute of Biomedical Sciences Abel Salazar (ICBAS), U.Porto, Department of Microscopy, Laboratory of
Histology and Embryology, R. Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal;
2
Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), U.Porto, Histomorphology,
Physiopathology and Applied Toxicology Group, R. dos Bragas 289, 4050-123 Porto, Portugal.
1
Total amounts of polycyclic aromatic hydrocarbons (PAHs) in the Oporto
district
MJ Rocha1,2; C Cruzeiro1,2; AS Caramelo1; JV Côrte-Real1; J Sousa2; E Rocha1,2
The evaluation of PAHs in sediments taken from the Douro River estuary,
Oporto district, revealed that this region has been highly impacted by these
compounds (Rocha et al. 2011). Because PAHs are extremely toxic, from aquatic
animals to humans, their assessment in surface waters of Oporto area has
become of the most interest. For this evaluation, during 2011, water samples
taken from Leça and Matosinhos (coastline) and from Fluvial and Freixo (Douro
River estuary) were analysed by gas chromatography – mass spectroscopy. All
analysed PAHs are included in the US Environmental Protection Agency (US
EPA) priority list, which includes acenaphthylene, acenaphthene, fluorene,
phenanthrene,
anthracene,
dibenz[a,h]anthracene,
fluoranthene,
pyrene,
indeno[1,2,3-cd]pyrene
and
benzo[k]-fluoranthene,
some
potentially
carcinogenic for humans; in particular benzo[a]anthracene, chrysene, benzo[b]
fluoranthene, benzo[a]pyrene and benzo[ghi]peryle-ne. Results, coming from
the analysis of these PAHs dissolved in water (DW) or adsorbed to suspended
particulate matter (SPM) revealed that the most polluted site was the Matosinhos
beach, where the average concentration of all PAHs was ≈ 62 ng/L (DW) and ≈
12 ng/L (SPM), respectively. The average amounts of the ∑16PAHs were, at the
coastline ≈ 57 ng/L (DW) and ≈ 11 ng/L (SPM), and at the Douro River estuary
≈ 48 ng/L (DW) and ≈ 9 ng/L (SPM). In this sense, since there is no information
about the possible lethal impacts of this environmental mixture of PAHs over
local fauna, a toxicity test using Artemia salina (ARC-test) will be done in order
to investigate this aspect. The latter data, together with those observed during
335
the chemical monitoring process will offer new insights about the impact of the
Spatial and annual distribution of estrogenic endocrine compounds in
evaluated PAHs in this region.
the Aveiro Lagoon
MJ Rocha1,2; C Cruzeiro1,2; AS Caramelo1; C Coutinho1; E Rocha1,2
Rocha et al. (2011). J Chromatogr Sci, 49(9), 695-701.
Acknowledgements
PTDC/MAR/70436/2006.
1Institute of Biomedical Sciences Abel Salazar (ICBAS), U.Porto, Department of Microscopy, Laboratory of
Histology and Embryology, R. Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal;
2Institute of Biomedical Sciences Abel Salazar (ICBAS), U.Porto, Department of Microscopy, Laboratory of
Histology and Embryology.;
3Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), U.Porto, Histomorphology,
Physiopathology and Applied Toxicology Group, R. dos Bragas 289, 4050-123 Porto, Portugal.
Ria de Aveiro is a very important area of the Portuguese coast, which has been
under an increasing anthropogenic pressure for several decades, a fact that has
contributed to the degradation of the water quality of this lagoon. Due to this
observation, this study reports the evaluation of seventeen endocrine disruptors
compounds (EDCs) from biological (estrone, E1and estradiol, E2), pharmaceutical
(ethinylestradiol, EE2), industrial and household (octylphenols, OPs; nonylphenols,
NPs; alkylphenols ethoxylates, APEOs; and bisphenol A, BPA) and vegetal
(formonometin, FORM; daidzein, DAI; genistein, GEN; biochanin A, BIO-A; and
sitosterol, SITO) origins. For this propose, water samples were collected during one
year, at eight sampling stations located into the lagoon and close to the sea. The
water samples were filtrated and all EDCs were extracted by solid phase extraction
(SPE) in Oasis HLB cartridges, cleaned in silica cartridges and the concentrations and
identities of the above referred pollutants were evaluated by gas chromatography –
mass spectroscopy. Briefly, the maximal amounts of these EDCs in the lagoon were:
E1≈ 23 ng/L; E2≈ 26 ng/L; EE2≈ 53 ng/L; ∑OPs≈ 83 ng/L; SNPs≈ 1,300 ng/L; ∑APEOs≈
14,600 ng/L; BPA≈ 300 ng/L; FORM≈ 10,000 ng/L; DAID≈ 300 ng/L; GEN≈ 160 ng/L;
BIO-A≈ 1,200 ng/L; SITO≈ 6,000 ng/L. Considering the literature data on effects on the
range of concentrations found, and normalizing the average amounts of the above
referred oestrogenic EDCs, in ethinylestradiol equivalents (EE2eq), we conclude that
endocrine disruption risk by oestrogenic exposure may exist in this area (EE2eq≈ 50
ng/L). Moreover, since there is no information about the possible lethal impacts of
337
this environmental mixture of EDCs over local fauna, a toxicity test using Artemia
Global amounts of pesticides and their toxic impacts in the Mondego
salina(ARC-test) will be done in order to observe this aspect. The latter data, together
River estuary
with those observed during the chemical monitoring process will contribute with
C Cruzeiro1,2; AS Caramelo1; J Soares 2; M Pardal3; M Reis3; E Rocha1,2; MJ Rocha1,2
new information about the impact of the evaluated EDCs in this region.
The Mondego River rises in the centre of Portugal, at the Mountain Range of
Serra da Estrela and reaches the Atlantic Ocean at Figueira da Foz. During its course
Acknowledgements
PTDC/MAR/70436/2006.
1Institute of Biomedical Sciences Abel Salazar (ICBAS), U.Porto, Department of Microscopy, Laboratory of
Histology and Embryology, R. Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal;
2Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), U.Porto, Histomorphology,
Physiopathology and Applied Toxicology Group, R. dos Bragas 289, 4050-123 Porto, Portugal.
of 227 km it runs through rural as well as highly urbanized and industrialized
areas before reaching its 1,600 ha estuary, where it becomes visibly polluted.
Its main pollution sources are waste waters, industries and the agricultural
run-off, end result of 15,000 ha of cultivated land (mostly rice and corn fields).
In this sense, the main objective of this study was to check for the presence
of 56 priority pesticides dissolved in surface water samples collected during
2010, from 7 sampling sites. Target compounds included several categories of
pesticides: insecticides, herbicides and fungicides. Analyzes were performed
using previously validated methods involving solid-phase extraction, in Oasis
HLB cartridges, followed by gas chromatography - mass spectroscopy (GC-MS/
MS). Results revealed that the maximal amounts of pesticides in the Mondego
waters were ∑insecticides≈ 11.6 μg/L, ∑herbicides≈ 2.8 μg/L and ∑fungicides≈
3.0 μg/L. Because there is no information about the possible lethal impacts of
this environmental mixture of pesticides over local fauna, a toxicity test using
Artemia salina (ARC-test) will be done in order to estimate this aspect. The latter
data, together with those observed during the chemical monitoring process will
provide new evidences to access the impact of the evaluated pesticides in the
biota of the region.
339
Acknowledgements
FCT SFRH/BD/79305/2011, PTDC/MAR/70436/2006, PEST-C/MAR/LA0015/2013
1Institute of Biomedical Sciences Abel Salazar (ICBAS), U.Porto, Department of Microscopy, Laboratory of
Histology and Embryology, R. Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal;
2Interdisciplinary Centre for Marine and Environmental Research (CIIMAR), U.Porto, Histomorphology,
Physiopathology and Applied Toxicology Group, R. dos Bragas 289, 4050-123 Porto, Portugal;
3Centre for Functional Ecology (CFC), Department of Life Sciences, Calçada Martim de Freitas, 3000-456
Coimbra, Portugal
Evolução do perfil físico-químico dos sedimentos do estuário do Rio Lima
J C O Lopes1; V Peixoto1; S Fernandes1
Ao longo do extenso litoral português existem extensas zonas húmidas de
grande importância ambiental e económica. Exemplos dessas zonas húmidas são os
sistemas lagunares costeiros designados por “Ria”, lagoas, e os sistemas estuarinos
como o do Lima. As zonas costeiras são, tipicamente, zonas de conflito de atividades,
sendo imperativo que se estabeleçam bases sólidas para a sua gestão integrada.
A monitorização dos sedimentos desempenha um papel fundamental em
estudos de contaminação pois estes atuam como um reservatório de contaminantes
e portanto como fonte destes contaminantes para a coluna de água e seres vivos.
Em Portugal, mais de 40% da população concentra-se na faixa litoral, o que
corresponde a 4 milhões de habitantes e, deste modo verifica-se uma elevada
pressão humana sobre a zona costeira e existem atualmente 13 estuários e sistemas
lagunares: a norte do Tejo (Minho, Cávado, Lima, Ave, Douro, Ria de Aveiro, Mondego,
Tejo) e a sul encontramos o Sado, Mira, Arade, Ria Formosa e Guadiana.
O estudo dos estuários manifesta um elevado grau de dificuldade uma vez que
estes sistemas aquáticos englobam geometrias e padrões de circulação hidrodinâmica
muito complexas e apresentam-se fortemente alterados por ações antropogénicas.
No presente trabalho pretendeu-se fazer a comparação entre a caracterização
físico-química sumária – pH, matéria orgânica, granulometria, teores em fósforo,
sódio e potássio – realizada em outubro/novembro de 2010 e os resultados obtidos
em outubro/novembro de 2014 dos sedimentos estuarinos do rio Lima.
As amostras em análise foram recolhidas em duplicado em 5 pontos de
amostragem distribuídos pelas duas margens do rio Lima: uma no parque náutico de
Darque, no Cabedelo, na Capela de S. Lourenço, na praia fluvial de Viana do Castelo e
341
no Parque da Cidade de Viana do Castelo.
Em 2010, os valores de pH variaram de 6,60 a 9,19; os valores de densidade aparente
Evolução do perfil físico-químico dos sedimentos do estuário do Rio Cávado
J C O Lopes1; V Peixoto1; S Fernandes1
variaram de 0,135 a 4,79 g/cm3; observaram-se valores de 0,07 a 2,32% para a matéria
orgânica tendo sido obtidos resultados entre 5,26 e 97,7 ppm de fósforo; foram ainda
É nas zonas costeiras onde se verifica o maior impacto da atividade humana,
registados valores mínimos de 76 e 22,8 ppm e máximos de 368 e 94,05 ppm para
pois os resíduos que atingem essas zonas são provenientes de fontes terrestres, de
o potássio e o sódio respetivamente. Em 2014 os valores de pH variaram de 7,00 a
drenagens dos rios e do lançamento de esgotos e ainda da geração de resíduos.
8,80; os valores de densidade aparente variaram de 0,99 a 1,57 g/cm3; observaram-se
Apenas o estuário do Cávado, na zona norte do Tejo, é aquele que hoje é
valores de 0,14 a 3,3% para a matéria orgânica tendo sido obtidos resultados entre 9,45
considerado uma zona protegida pois é constituído por espécies representativas das
e 159,04 ppm de fósforo; foram ainda registados valores mínimos de 56,56 e 308,38
comunidades vegetais do litoral ibérico com relativa abundância e, inclusivamente,
ppm e máximos de 353,28 e 772,08 ppm para o potássio e o sódio respetivamente.
por algumas espécies ameaçadas, justifica a manutenção das medidas de proteção
Observou-se um aumento significativo nos teores de fósforo nos sedimentos bem
como nos níveis de sódio.
que estiveram na origem da criação daquela Área Protegida e que resultou
posteriormente na criação do Parque Natural do Litoral Norte.
No presente trabalho pretendeu-se avaliar a evolução do perfil físico-químico
sumário – pH, matéria orgânica, granulometria, teores em fósforo, potássio, cálcio,
Instituto Politécnico de Viana do Castelo - Escola Superior Agrária
1
magnésio e sódio – dos sedimentos estuarinos quantificando ainda o grau de
contaminação com crómio e cádmio (muitas vezes associados a efluentes da indústria
têxtil predominante ao longo do curso do rio Cávado) através da comparação entre
os resultados obtidos em outubro/novembro de 2010 e os resultados obtidos em
outubro/novembro de 2014.
As amostras em análise foram recolhidas em duplicado em 5 pontos de
amostragem: três das recolhas realizaram-se na margem norte distribuídas entre a
foz do rio Cávado e a ponte de Fão e outras duas, na margem sul, uma debaixo da
ponte de Fão e a outra junto ao clube náutico.
Os resultados obtidos demonstraram uma maior densidade nas amostras
recolhidas na zona estuarina. Nas amostras recolhidas em 2010, os valores de
pH variaram de 6,47 a 8,01; os valores de condutividade variaram de 0,78 a 5,98
(amostras junto ao estuário) o que evidencia um efeito acentuado da água do mar
343
nesta zona; para os valores de fósforo foram obtidos resultados entre 62 ppm e 210
Caracterização físico-química dos sedimentos do estuário do Rio Minho
ppm; foram ainda registados valores mínimos de 8,66 e 14,03 e máximos de 82,24 e
J C O Lopes1; V Peixoto1; S Fernandes1
264,3 ppm para o potássio e o cálcio respetivamente. Quanto ao sódio os pontos de
amostragem apresentaram níveis elevados (> 1000 ppm). No que refere aos metais
As regiões litorais, nas últimas décadas têm sido caracterizadas por um elevado
pesados analisados, o cádmio apresentou-se abaixo dos limites legais, no entanto, o
crescimento populacional e urbanístico que foi auxiliado pelo rápido desenvolvimento
crómio apresentava valores acima dos limites legais em vigor.
industrial, no qual se traduz numa diminuição da qualidade ambiental das zonas
Nas amostras recolhidas em 2014, os valores de pH variaram de 6,80 a 9,00; os
costeiras.
valores de condutividade variaram de 0,05 a 0,70 (amostras junto ao estuário) o que
Em Portugal, mais de 40% da população concentra-se na faixa litoral, o que
evidencia um efeito acentuado da água do mar nesta zona; para os valores de fósforo
corresponde a 4 milhões de habitantes e, deste modo verifica-se uma elevada pressão
foram obtidos resultados entre 8,53 ppm e 64,55 ppm; foram ainda registados
humana sobre a zona costeira e existem atualmente 13 estuários e sistemas lagunares:
valores mínimos de 34 e 5 e máximos de 73,96 e 317 ppm para o potássio e o cálcio
a norte do Tejo (Minho, Cávado, Lima, Ave, Douro, Ria de Aveiro, Mondego, Tejo) e a sul
respetivamente.
encontramos o Sado, Mira, Arade, Ria Formosa e Guadiana.
No que refere aos metais pesados analisados, o cádmio apresentou-se abaixo
No presente trabalho pretendeu-se caracterizar o perfil físico-químico sumário –
dos limites de deteção, no entanto, o crómio continua a apresentar valores elevados
pH, matéria orgânica, granulometria, teores em fósforo, potássio, cálcio, magnésio e
(máximos acima de 300 ppm).
sódio – dos sedimentos estuarinos quantificando ainda o grau de contaminação com
crómio e cádmio.
As amostras em análise foram recolhidas em duplicado em 5 pontos de amostragem
Instituto Politécnico de Viana do Castelo - Escola Superior Agrária
1
na zona estuarina do rio Minho, na margem Portuguesa.
Nas amostras recolhidas os valores de pH variaram de 8,1 a 8,6; os valores de
condutividade variaram de 0,13 a 0,54; para os valores de fósforo foram obtidos
resultados entre 6,38 ppm e 18,07 ppm; foram ainda registados valores mínimos de
21,24 e 56 ppm e máximos de 33,28 e 866 ppm para o potássio e o cálcio respetivamente.
Quanto ao sódio os pontos de amostragem apresentaram níveis superiores a 200 ppm.
No que refere aos metais pesados analisados, o cádmio apresentou-se abaixo dos
limites de deteção e o crómio apresentava valores superiores a 0,260 ppm.
Instituto Politécnico de Viana do Castelo - Escola Superior Agrária
1
345
Diseño de un índice regional de calidad del agua para embalses basado
en la lógica difusa
Jacinto Elías Sedeño Díaz1; Eugenia López López2
El objetivo de este trabajo es proponer un índice de calidad del agua que
sea aplicable a embalses de una misma cuenca utilizando la lógica difusa. Ésta
contribuye a librar la frontera de los valores Crisp tales como los límites máximos
permisibles que establecen los marcos legales, ofreciendo una alternativa más real
en la integración de indicadores de calidad del agua de naturaleza y dimensiones
diferentes. La mayoría de los índices de calidad del agua han sido diseñados para
su aplicación en ríos y corrientes y basados en la opinión de expertos; sin embargo,
cuando el agua es embalsada su calidad sufre cambios generalmente en deterioro
de la misma. México es un país con muchos contrastes en cantidad y calidad del
agua. La Mesa Central es una región con alta densidad poblacional, escasos recursos
hídricos y alto impacto incidiendo sobre éstos. La gestión de recursos hídricos en
la cuenca del río Lerma se logra con un sistema de embalses artificiales donde se
almacena el agua con el fin de satisfacer las necesidades humanas en la cuenca, tales
como el abastecimiento público, la industria, la agricultura y la recreación, entre otros.
Por lo anterior, se tiene la necesidad de contar con un sistema de evaluación que
permita tener un diagnóstico rápido y económico que ofrezca un score cuantificable
y confiable sobre la calidad del agua de estos embalses. Se realizó un monitoreo
sobre los 10 principales embalses de la cuenca del río Lerma para caracterizar su
calidad del agua en un ciclo anual. A partir de los datos generados, se seleccionaron
los indicadores de calidad del agua que no representaran redundancia y fueron
modelados en un universo de discurso en una escala de cero a cien (Funciones de
Pertenencia) con base en el gradiente de concentraciones que presentaron en los
347
diferentes embalses. Así, el índice propuesto utiliza ocho indicadores de calidad del
Natural radiation and geochemical data of the Vila Pouca de Aguiar massif,
agua (oxígeno disuelto, conductividad, nitratos, transparencia del disco de Secchi,
northern Portugal
color, ortofosfatos, nitrógeno amoniacal y temperatura). Cada indicador cuenta con
L. Martins1; L. Martins1; M. Gomes1; R. Teixeira1; A. Miranda1; A.J.C.S. Pereira2; L.J.P.F. Neves2
su función de pertenencia. El Sistema Mandami de Inferencia Fuzzy se compone
por 633 reglas que son defusificadas por el método LOM (Large of Maximum), para
dar un score de 0 a 100, con siete categorias verbales: Excelente, Buena Calidad,
Radon gas has a high mobility in natural systems and it is important to diagnose
the radiological impact of rocks and soils in the concentrations of indoor radon.
Ligeramente Contaminada, Calidad Regular, Contaminada, Agua Muy Contaminada
In Vila Pouca de Aguiar region outcrops a post-tectonic granite massif with an
e Inaceptable. El índice fue validado por comparación con un índice de calidad del
elongated form, constrained by NNE-SSW fractures, where three types of biotite
agua que utiliza 13 indicadores, y su uso a través de la cuenca se probó por aplicación
granites occur (Souto, Telões and Pedras Salgadas). Two of these show different stages
del mismo en cinco cuerpos de agua adicionales. Los embalses fueron categorizados
of alteration (episienitization). The natural radiometric background was measured in
desde excelente hasta calidad regular, lo cual estuvo relacionado con su ubicación.
84 locations, using a portable gamma-ray spectrometer. Similar U and Th contents
Aquellos con mejor calidad del agua se localizaron en las zonas altas con menor
have been found in the Pedras Salgadas granite (U= 7 to 21 ppm and Th= 19 to 38
densidad poblacional y los de menor calidad en zonas bajas con mayor influencia
ppm) and in the altered facies (U= 6 to 21 ppm and Th= 24 to 32 ppm). There is also U
humana.
enrichment in the Telões granite with reddish tones. The Souto granite has the least
radiogenic profile of U and Th, although all these values are higher than the crustal
1Instituto Politécnico Nacional, Coordinación Politécnica para la Sustentabilidad;
2Instituto Politécnico Nacional, Escuela Nacional de Ciencias Biológicas
average (average of K- 2.1%; U- 2.7 ppm Th- 9.6 ppm). Concerning the external
radiation exposure, the average values of all studied granites were slightly lower than
the common granitic rocks of ZCI (ca. 200 ηGy/h). SEM studies indicate the presence
of U and Th in radioactive accessory minerals, especially zircon, thorite and allanite
in the Telões granite; monazite, xenotime and zircon in Pedras Salgadas and Souto
granites, including thorite in the latter. The geometric mean of radon activity in 91
dwellings are higher (568 Bq/m3) than the limit of 400 Bq/m3 defined by the national
legislation on indoor air quality, and subsequently there is a significant proportion
of dwellings that exceed this limit (n= 57). The concentrations of the radionuclides
in the rocks show a good correlation with indoor radon measured in dwellings, so
it can be conclude that the granites of Telões and Pedras Salgadas, including their
weathering stages, are more radiogenic than the Souto granite.
349
1Universidade de Trás-os-Montes e Alto Douro, UTAD, Quinta de Prados, 5000-801 Vila Real, Portugal, and
CEMUC;
2CEMUC Department. of Earth Sciences, University of Coimbra, 3000-272 Coimbra; Portugal
Monitoring the impacts on the ecosystem integrity of Portelo stream
(Douro basin, NE Portugal) after a large spill of mining wastes
Amílcar Teixeira1; C. Fernandes1; A. Geraldes1; S. Varandas2
After a period of intense precipitation, in December 2009, a big amount of
wastes from an abandoned mine were spilled into Portelo stream, a tributary of
Rio Sabor (Douro basin)located in the Montesinho Natural Park (NE Portugal). The
wastes covered the riverbed near the mine and some riparian and agricultural
downstream areas. From 2010 to 2014 we monitored the impacts of waste spill
on the ecosystem integrity, namely in the water chemistry, channel and riparian
habitats and on the composition of macroinvertebrate communities. Eight
sampling points were selected along the hydric system. Toxicity experiments
were developed in laboratory for one fish species, the Northern Iberian barbel
(Luciobarbus bocagei), an endemic cyprinid present in the River Sabor. The
results showed a big increase of conductivity (>300 µS.cm-1), total suspended
solids (> 100 mg.L-1) and a drastic decrease of pH (<5) in the water analyses. It
was also detected higher concentrations of copper, aluminium and cobalt in the
water (52 physical and chemical parameters were determined, including different
heavy metals). Aquatic and riparian habitats were severely disturbed and several
metrics (e.g. taxonomic richness, diversity, evenness) confirmed biotic substantial
changes in composition and abundance of macroinvertebrate communities.
Furthermore, biochemical indicators (Na+and K+ plasmatic concentrations)
used in laboratory tests showed to be sensitive, under acute copper exposure of
barbel populations (different copper concentrations, 0.06-0.48 ppm were used),
and contributed to justify the disappearance and or reduction of fish species in
the affected located in the Montesinho Natural Park. Rehabilitation of disturbed
351
areas must be implemented in order to recover the ecological integrity of the
Sediment quality assessment related to benthic macrofauna in São
aquatic and riparian ecosystems.
Francisco River (Minas Gerais, Brazil)
Esther Bautista Chamizo1; I. Riba2; M.J. Salamanca2; V. Almagro-Pastor2; K.C. Pereira2; A. Luque3; T.
A. DelValls2
1CIMO-ESA-IPB - Mountain Research Centre, School of Agriculture, Polytechnic Institute of Bragança;
2CITAB - Centre for the Research and Technology of Agro-Environmental and Biological Sciences, University
of Trás-os-Montes e Alto Douro
The São Francisco River (SFR) is one of the most important Brazilian and South
American rivers. However, there are plenty sources of pollutants along this river, and
there are a few environmental assessments developed in this area. Therefore, a weightof-evidence (WOE) approach was applied to locate and identify the pollution sources
in the SFR course surrounding the Votorantim Metais-Três Marias refining factory.
This WOE approach was based on two different lines of evidence (LOEs): the structure
of communities (Biological Monitoring Working Party score system, abundance of
taxa, number of individuals, Margalef´s species richness, Pielou´s evenness, and
Shannon-Wiener diversity), and some physico-chemical determinations in sediments
(percentage of fines, content of Total Organic Carbon, nitrate, ammonium, ammonia,
ammoniacal nitrogen, Al, As, Fe, and Simultaneously Extracted Metals/Acid-Volatile
Sulfides. As a result, the Três Marias Wastewater Treatment Plant (WWTP) was the most
important source of pollution of this stretch of the SFR. Nevertheless, the Votorantim
Metais-Três Marias factory also showed contamination that affected downstream of the
River course. Therefore, this paper recommends a stronger control of the effluents on
the river, besides more measures allocated to reduce the contaminant emission from
the WWTP. Besides, three different sentinel species (Tanytarsus sp., Crytochironomus
sp. and Polypedilum sp.) were identified to facilitate future monitoring assessments.
1Universidad de Cádiz;
2UNESCO UNITWIN/UNICOP, Faculty of Environmental and Sea Sciences, University of Cádiz;
3Departamento de Biología,Universidad de Las Palmas de Gran Canaria
353
Estudios preliminares de evaluación de toxicidad del fenitrotión en dorada (Sparus aurata)
J. Arellano1,3, G. Albendín1, P. Padilla1, F. Soto-Mancera1, C. Sarasquete2,3, M.P. Mánuel-Vez 4, M.I.
Arufe1,3
Los insecticidas organofosforados son inhibidores de la enzima acetilcolinesterasa,
provocan una acumulación excesiva del neurotransmisor acetilcolina e interrumpen la
transferencia del impulso nervioso. Esta enzima es crítica para el funcionamiento normal
del sistema nervioso, de la fisiología, del comportamiento, de la reproducción y de la
supervivencia. Asimismo, la exposición a estos insecticidas puede provocar alteraciones
morfológicas en el hígado, digestivo y branquias de los organismos. El presente trabajo
tuvo como objetivo la evaluación de la toxicidad del insecticida organofosforado fenitrotión
en larvas de dorada (Sparus aurata). Se llevaron a cabo estudios de toxicidad estáticos con
renovación durante 72 h registrando la supervivencia de las larvas y la inhibición de la AChE.
La concentración letal para el 50% de los individuos (LC50) fue de 1,027 mg/L (intervalo de
confianza del 95%= 0,706-1,34 mg/L), por lo que este compuesto resulta moderadamente
tóxico para las larvas estudiadas. El porcentaje de inhibición de la actividad colinesterasa
aumentaba a mayor concentración de fenitrotión. La concentración más alta a la que se
midió la actividad colinesterasa (2,5 mg/L) mostró un 58% de inhibición de esta enzima en
larvas endotróficas de dorada (Sparus aurata) expuestas durante 72 h.
Laboratorio de Toxicología. Facultad de Ciencias del Mar y Ambientales. Universidad de Cádiz. Polígono
Río San Pedro s/n, Puerto Real, 11510 Cádiz (España).
2
Instituto de Ciencias Marinas de Andalucía (ICMAN). Consejo Superior de Investigaciones Científicas
(CSIC). Polígono Río San Pedro s/n, Puerto Real, 11510 Cádiz (España).
3
Unidad Asociada de Calidad Ambiental y Patología (CSIC-UCA)
4
Departamento de Química Analítica. Facultad de Ciencias del Mar y Ambientales. Universidad de Cádiz.
Polígono Río San Pedro s/n, Puerto Real, 11510 Cádiz (España).
e-mail: juana.arellano@uca.es
1
355
Ecosystem Level Effects
Diversidad y dinámica planctónica (fitoplancton y rotíferos) y la producción primaria de un lago urbano hipertrófico de la Ciudad de México.
J L, Gama-Flores1; E, Huidobro-Salas1; A, Rodriguez-Rocha1; SSS, Sarma1; N,Sarma1
El Plancton es un componente importante de las comunidades acuáticas, cuyo
estudio permite observar su estructura, biomasa y dinámica así como los cambios
espacio temporales que se producen en ellas por factores naturales y antrópicos.
De particular relevancia resulta estudiar dichas comunidades en cuerpos urbanos
eutrófico-hipertróficos, y con recambio de agua residual tratada. En este estudio
se presenta la diversidad, la biomasa, y la dinámica del fitoplancton y zooplancton
del lago de Tezozomoc, México, D.F, durante la estación de verano. Para cubrir
estos objetivos se realizaron muestreos litorales de tres micro-ambientes del
sistema durante tres meses, dos veces por semana, y se caracterizó físico, química y
nutrimentalmente (temperatura, oxígeno disuelto, pH, alcalinidad, N : P y la cantidad
de clorofila a), y la producción de cada sitio simultáneamente. Se encontraron
28 especies, siendo Chlorophyceae, Euglenophyceae y Cyanophyceae las clases
taxonómicas predominantes, con mayor número de especies. En cuanto a la
abundancia específica, se observó una alternancia entre los grupos de Scenedesmus
spp,Eglena gracilis, E viridis, Microcystis bothrys quienes dominaron inicialmente en
tanto que Pediastrum boryanum, Golenkinia radiata, Pandorina elegans, Anabaena
spiroides, lo fueron posteriormente. En el caso del zooplancton (los rotíferos), se
encontraron 28 especies siendo Brachionidae la más diversa. Para la comunidad
de rotíferos, también se registró el comportamiento de alternancia de grupos
predominantes, siendo las especies de la familia Brachionidae dominantes de inicio,
357
y posteriormente reemplazada por las especies de la familia Trichocercidae. Este
Eutrophication due to sewage discharges causes blooms of Peridinium quin-
comportamiento fue según el microambiente del sistema. Los valores de diversidad
quecorne (Dinophyceae) off the coasts of the Port of Veracruz, Mexico: A case
de Shannon- Wiener fueron de medio alto, para ambos grupos de planctontes, lo
Alfredo Pérez Morales1; J. A. Aké-Castillo1; G. Campos-Bautista2
mismo que la biomasa y producción primaria del fitoplancton. Esta dinámica de
ambas comunidades fue en respuesta a un medio muy alcalino, con una relación N:P
< 1, denotando una limitación muy grande nitrógeno y oxigenación variable.
The Port of Veracruz is the most important commercial seaport in Mexico,
with a population of over 550,000 inhabitants, is a growing city. The increase in
human population and the inadequate management of the urban growth are
the principal factors that influence the saturation of residual water treatment
Universidad Nacional Autonoma de Mexico
1
plants; these situations provoke the release of raw sewage to the sea, increasing
nutrients and turn available to several phytoplankton species. In this regard, we
perform a monthly monitoring during five years (2010-2014), in a station located
on the north side of town in front of the main waste outlet of the residual water
treatment plant. In this station, we evaluated physicochemical factors such as
temperature, salinity, pH and oxygen as well as nutrients such as ammonium
(NH4), nitrites (NO2), nitrates (NO3), phosphates (PO4) and silicates (SiO2), also
were analyzed water samples for identification of microalgal associations. Our
results showed high variations in physicochemical factors: temperature from
19 to 32 °C, salinity from 21.5 to 36.5 ups, pH from 7.4 to 9 and oxygen from
2 to 11.5 mg/L; as well as high variations of nutrients: NH4 from 0.02 to 16.4
mg/L, NO2 from 0.002 to 0.22 mg/L, NO3 from 0.8 to 5.6 mg/L, PO4 from 0.12
to 7.85 and SiO3 from 0.02 to 24.5. These conditions have favored some bloomforming microalgae species, mainly of the genera Chaetoceros, Thalassiosira,
Skeletonema, Gymnodinium and Pseudo-nitzschia. However, the most
dominant and recurrent blooms are the non-toxic dinoflagellate Peridinium
quinquecorne, with concentrations exceeding 4 x 108 cells/L. These blooms
have been observed mainly in the warmer months (April to September). These
359
results indicated that release of raw sewage to the sea may provoke changes in
ecosystem structure by direct effect.
Emerging Toxic Compounds
Ecotoxicity of diltiazem
Natalia Ros1; Eduardo Perales1; Cristina B. García1; Mª Pilar Ribate1 ; Beatriz Giner1; Laura Lomba1
Instituto de Ciencias Marinas y Pesquerías;
Acuario de Veracruz A.C.
1
2
In last years, an important pharmaceutical development has been produced. The
presence of the Pharmaceutical Active Compounds (PhAcs) in the environment has
become a recent topic because of its presence in aquatic ecosystem. However, there
are no enough studies about what happen with these chemicals in the environment
and how affect in nature, though the main problems are related to their stability,
persistence and the action on many organism.
In this work, we present a part of our research project which is based on the
ecotoxicological study of drugs; in this case diltiazem which is calcium channel
blocker used in hypertension treatment. To carry out this study, different bioasays
have been used. Firstly, the standardized toxicity test using Vibrio fischeri exhibited
during 30 minutes and secondly, the Daphnia magna acute inmobilization test in
accordance with the OECD 202 test conditions and operating protocol during 24
hours. Finally, a critical aggregation concentration has been obtained for studied
compound due to the relationship between the bioavailability and ecotoxicity.
From the experimental results, we can conclude that this drug (diltiazem), taking
into account the Passino and Smith classification, can be considered as practically
harmless for Vibrio fischeri and Daphnia magna.
GreenLife Research Group. Faculty of Health Sciences. San Jorge University. Villanueva de Gállego,
Zaragoza, Spain
1
361
Ecotoxicology study of a mixture of drugs
Estefanía Zuriaga1; Laura Lomba1; Beatriz Germán1; Pilar Mª Lanuza1; Luis Aldea1; Natalia Ros1
Despite their impact on the environment remains unknown, the presence of
a large number of pharmaceuticals in fresh water is recognized as an important
environmental problem (Fent et al., 2006). In this study we have analyzed the effect
of a mixture of pharmaceuticals in order to evaluate the environmental risk of the
presence of these pharmaceuticals in the environment. The chosen pharmaceuticals
are ibuprofen and omeprazole.
The ecotoxicity of these two pharmaceuticals and their mixture has been evaluated
using Vibrio fischeri as bioindicator. Synergistic effects were observed when
combinations of ibuprofen and omeprazole were tested. Concentration addition and
independent action methods have been used to predict the ecotoxicological effect
of the studied mixtures. In both cases, an overestimation of the actual ecotoxic effect
of the mixture has been found. Finally, the toxic effect has been also tested after 5
days.
This study provides an example of the ‘effect-driven approach’ described by Escher
et al.(Escher and Fenner, 2011) for determining the risks associated with pollutant
transformation products. According to this approach, compounds are prioritised for
transformation product isolation, purification, and further study if the reaction mixture
increases in toxicity in parallel with decreasing parent compound concentration.
In this case, the effect driven approach has been applied and we can conclude that
transformation products of the studied drugs should be prioritized for risk assessment
purposes.
Escher, B. I., and K. Fenner, 2011, Recent Advances in Environmental Risk Assessment of Transformation Products:
363
Environmental Science & Technology, v. 45, p. 3835-3847.
Fent, K., A. A. Weston, and D. Caminada, 2006, Ecotoxicology of human pharmaceuticals: Aquatic Toxicology, v. 76,
p. 122-159.
GreenLife Research Group. Faculty of Health Sciences. San Jorge University. Villanueva de Gállego, Zaragoza,
Spain
1
Chronic effects in crustacean Daphnia magna after exposure to a solution
of carbamazepine under single catalytic (TIO2) ozonation.
A.L. Oropesa¹*; F.J., Beltrán² ; A.M., Floro³; J., Guerreiro³; J.J.P., Sagasti²; P., Palma3,4
Advanced oxidation processes are used to remove pharmaceuticals from water.
The aim of the present study was evaluate the toxicological efficiency of a single
catalytic ozonation treatment (TiO2), in the elimination of carbamazepine (CBZ), an
antiepileptic drug, through chronic endpoints. Therefore, in this study a clean waster
sample was spiked with 200 µgL-1 of CBZ and submitted to the treatment, in order
to eliminate the drug up to 5 μgL-1. A dilution series of the obtained solution and its
mixture with fenoxycarb (1 μgL-1 FEN; an insect growth regulator that mimics the
action of methylfarnesoate) were performed (25, 50 and 100% (v/v) in ASTM; 25%+1
μgL-1 FEN, 50%+1 μgL-1 FEN and 100%+1 μgL-1 FEN (v/v) in ASTM)to evaluate the
possible effects on the life cycle and sex differentiation of Daphnia magna. The
endpoints used were fecundity (number of offspring per female), percentage of
male production, developmental abnormalities and growth of females, through the
development of the chronic reproduction bioassay. Further, the possible anti-juvenoid
activity of the drug was assess in the presence of fenoxycarb (FEN, 1 µgL-1 positive
control), an insect growth regulator, that mimics the action of methylfarnesoate and
promoted the production of 95% of male offspring at the given concentration.
All solutions (100%, 50%, 25%) and mixtures (100%+FEN, 50%+FEN, 25%+FEN)
caused an important decrease in the number of offspring produced per female
compared to the control groups (ASTM+DMSO). Females from 100% solution died
throughout the test and produced high number of malformed and dead offsprings.
The size of females decreased in the mixtures comparing to the control groups. On
the other hand, production of males has been observed only in the mixtures.
365
The chemical efficiency of this technology for removal the CBZ from water
Physiological alterations on the marine microalga Tetraselmis suecica
has already been demonstrated; nonetheless the toxicological efficiency
exposed to the uv filter benzophenone-3
seemscommittedprobably due to the hydrogen peroxide production (reactive
Marta Seoane1; Marta Esperanza1; Carmen Rioboo1; Concepción Herrero1; Ángeles Cid1
oxygen species) in the final treated solution.
The development of new highly sensitive analytical techniques has revealed the
Authors wish to thanks the financial support given to A.L. Oropesa by Ministerio
presence of large amount of chemical compounds in the aquatic environment. These
de Educación, Cultura y Deporte within the Programa Estatal de Promoción del
substances are known as emerging pollutants and the presence and the awareness
Talento y su Empleabilidad en I+D+i, Subprograma Estatal de Movilidad, del Plan
of their potential environmental impacts have become evident in recent years.
Estatal de Investigación Científica y Técnica y de Innovación 2013-2016 through
Due to the growing concern about the harmful effects of exposure to ultraviolet
a post-doctoral research grant (CAS14-00224), and also to “Consejería de Empleo,
radiation, the use of UV filters has increased in all kind of cosmetic products,
Empresa e Innovación of Gobierno de Extremadura” and the Feder funds for the
becoming emerging pollutants. Sunscreens enter the marine environment mainly
economic support. Also, acknowledgement is given to MINECO of Spain and Feder
directly from the skin, being their presence particularly strong in coastal waters
Funds for economic support through project CTQ2012/35789/C02/02.
where aquatic recreational activities were carried out, mostly in summer season;
as well as indirectly through wastewater discharges or river inputs. Unlike priority
Unidad de Toxicología. Departamento de Sanidad Animal. Facultad de Ciencias. Universidad de
Extremadura, Badajoz 06071, España;
2Departamento de Ingeniería Química y Química Física. Facultad de Ciencias. Universidad de Extremadura,
Badajoz 06071, España;
3Departamento de Tecnologias e Ciências Aplicadas. Escola Superior Agrária de Beja, Beja 7801-295,
Portugal;
4Departamento de Tecnologias e Ciências Aplicadas. Escola Superior Agrária de Beja, Beja 7801-295,
Portugal. CIMA-Centro de Investigação Marinha e Ambiental, CIMA, FCT, Universidade do Algarve, Faro
8005-139, Portugal
1
pollutants, these compounds are not regulated and their potential ecotoxicological
effects are unknown.
Among sunscreens, benzophenone-3 (BP-3) is one of the organic UV filters
most used and detected in coastal waters. The aim of this study was to determine
the potential toxic effects of increasing concentrations of BP-3, ranged from
0.25 to 2 mg L-1, on the marine microalga Tetraselmis suecica, at level of various
physiological parameters. After 24 h of exposure to the contaminant, several
cytotoxicity biomarkers were analysed by flow cytometry. Assayed parameters were
chlorophyll a autofluorescence, cell viability, metabolic activity, intracellular pH and
cytoplasmic and mitochondrial membrane potentials. Cells remained viable in the
presence of BP-3; however, photosynthesis and intracellular pH were affected at
the highest concentration assayed. Furthermore, metabolic activity increased in a
367
concentration-dependent manner and cytoplasmic and mitochondrial membranes
Effects of psychopharmaceuticals exposure on RNA/DNA ratios in fish species
were depolarised respect to control cells. These results suggest that the presence
S.P Machado1; M. Araújo1,2; M. Ferreira1; S. Ramos
of this UV filter in the environment could damage the phytoplankton community
causing a misbalance in marine ecosystems.
The widespread detection of a large variety of pharmaceuticals in environment
has raised concern about the potential ecological impact of these bioactive
substances. The potential to elicit unwanted responses in non-target species, such as
Laboratorio de Microbiología, Facultad de Ciencias, Universidad de A Coruña. Campus de A Zapateira s/n
fish, even at low concentrations is recognized and risks must be assessed. The ratio of
15071 A Coruña
RNA/DNA measured in fish muscle is an important ecophysiological indicator of fish
1
general condition and nutrition status. The aim of this study was to evaluate RNA/
DNA ratio in juvenile European seabass (Dicentrarchus labrax) after chronic exposure
to psychopharmaceuticals (PPs). Fish (average weight 9 g in the 1st assay and 67 g in
the 2nd assay) were exposed separately to two different PPs: fluoxetine, a selective
serotonin reuptake inhibitor at 0.05 and 5 µg/L and to venlafaxine, a serotonin
norepinephrine reuptake inhibitor at 0.01 and 1 µg/L. Both chronic exposures
endured for 21 days, followed by 7 days of an exposure-free period to assess
reversibility of effects. Similarly the RNA/DNA ratio will also be evaluated in wild fish
species, seabass and flounder (Platichthys flesus), collected from Douro and Lima
estuaries. Preliminary results obtained indicate the presence of both PPs in water of
these estuaries. Our laboratory exposure results suggested that PPs have an impact
in fish physiological condition,leading to changes in the ratios between exposed
and unexposed fish. RNA/DNA ratio increased greatly after 1 day of fluoxetine and
venlafaxine exposure followed by decrease at day 21. A steady recuperation after
the 7 days of the exposure-free periodwas observed for fluoxetine. Hence, exposure
to PPs alters RNA/DNA ratio, highlighting its potential as a reliable biomarker of
exposure to emerging contaminants.
369
Acknowledgements
This study was supported by FCT project PTDC/AAG-MAA/2405/2012 and Pest-C/MAR/LA0015/2013.
Keywords
Psychopharmaceuticals, fish, muscle, RNA/DNA ratio
METOX, CIMAR/CIIMAR – Interdisciplinary Centre of Marine and Environmental Research, University of
Porto, Rua dos Bragas, 289, 4050-123 Porto, Portugal;
2UTAD – University of Trás-os-Montes and Alto Douro, Apartado 1013, 5001-801 Vila Real, Portugal;
3Ecobiotec, CIMAR/CIIMAR – Interdisciplinary Centre of Marine and Environmental Research, University of
Porto, Rua dos Bragas, 289, 4050-123 Porto, Portugal.
1
Effects of antidepressant fluoxetine in European sea bass juvenile´s behavior
B. Pinto1,2; S. Ramos3; M. Ferreira2
Recently, the presence of several pharmaceuticals as antidepressant fluoxetine (FX)
and their metabolites has been documented in aquatic ecosystems. FX is a selective
serotonin reuptake inhibitor that operates in the serotoninergic system of the central
nervous system. These compounds are not always eliminated in wastewater treatment
plants, reaching aquatic ecosystems as rivers and estuaries. The study aims to evaluate
the effect of FX exposure on European sea bass (Dicentrarchus labrax) juveniles at the
behavior level and also at the expression of associated specific brain genes. For this
purpose, an exposure assay of 21 days plus 7 days of recuperation was conducted. To
evaluate potential changes of juveniles behavior, points of analysis were selected (Day
1, 3, 7, 15, 21, 28). Swimming velocity was evaluated using a raceway device, registering
the time that each individual took to swim through 3m of the raceway. Anxiety was
also assessed by scototaxis (dark/light preference), observing the preference of each
juvenile for the black or the white compartment from colorless central zone of an
aquarium. The preference of black compartment was used as proxy of anxiety. Raceway
results showed that only exposed fishes to the highest FX concentrations (5 and 50
µg/L) swam throughout the raceway, with an average swimming velocity of 0.26 ± 0.18
m s -1. Scototaxis results denoted a decrease of anxiety in exposed fishes from day
15 until the end of the exposure, with more than 50% of juveniles avoiding the black
compartment. Further studies will analyze the expression of seven associated genes
from brain tissue samples collected on the 21th and 28th day. The behavioral and
gene expression results will be interpreted and discussed in the context of potential
contamination risk associated with contamination by psycho pharmaceuticals.
371
Acknowledgements
This study was supported by FCT through the project PTDC/AAG-MAA/2405/2012 and Pest-C/MAR/
LA0015/2013
FCUP - Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre, 4169-007 Porto
METOX, CIMAR/CIIMAR - Centro Interdisciplinar de Investigação Marinha e Ambiental, Rua dos Bragas, n.
289, 4050-123 Porto.
3
Ecobiotec, CIMAR/CIIMAR - Centro Interdisciplinar de Investigação Marinha e Ambiental, Rua dos Bragas,
n. 289, 4050-123 Porto;
Polystyrene microplastic localization and distribution in the gills and digestive gland of mussel Mytilus galloprovincialis
L. Garmendia1; B. Zaldibar1; U. Izagirre1; E. Bilbao1; M. Irazola2; S. Danielsson3; A. Bignert3; N. Etxebarria2; M. Soto1; I. Marigómez1
1
2
Plastics are present in many aspect of every day’s life due to their special
properties (versatility, lightweight, hardness, durability and production costs). The
annual world production of plastic has risen up to 288 million tons in 2012. It has
been estimated that about 10% of the plastics end up in the sea. Both manufactured
microplastics (< 5 mm) and those yields from the fragmentation of larger plastics
once they are exposed to degrading agents, can be accumulated in the biota and
produce negative effects. Within this context, the aim of the present work is to set up
a possible protocol in order to detect microplastics and to find out the accumulation
patterns of polystyrene microplastics in the soft tissues of mussels at laboratory
conditions. In order to achieve these objectives mussels, Mytilus galloprovincialis
were collected in a clean area of the Basque coast and exposed for 1, 4, 8 and 72
hours to three different sizes (6, 10 and 90 µm diameter) of polystyrene microplastics
in seawater. Different organs, including gills and digestive gland were collected and
then, the observation of the polystyrene microplastics was performed in different
ways including FT-IR, Raman spectroscopy, polarized light microscopy, transmission
electron microscopy and optical microscopy. Preliminary results indicated that FT-IR
and polarized light microscopy are very useful tools to observe microplastics in the
soft tissues of mussels. Regarding size distribution, 90 µm microplastics do hardly
penetrate in the mussel tissues since they are mainly localized in the gut. In contrast,
smaller (6 and 10 µm) microplastics are observed in the connective tissue of the
digestive gland after 4 h of exposure.
Acknowledgements
373
Funded by the Swedish Museum of Natural History, University of Stockholm, Sweden and the University of
the Basque Country/Basque Government through a grant to Consolidated Research Groups (Consolidated
Res Grp IT810-13).
Cell Biology and Environmental Toxicology Research Group, Research Centre for Experimental Marine
Biology and Biotechnology (PIE); Zoology and Animal Cell Biology Department (Faculty of Science and
Technology), University of the Basque Country, P.O. Box 6;
2
Ikerkuntza eta Berrikuntza Analitikoa Research Group. Analytical Chemistry Department (Faculty of Science
and Technology), University of the Basque Country, P.O. Box 644, E-48080 Bilbo, Basque Country, Spain;
3
Swedish Museum of Natural History, University of Stockholm, P.O. Box 50007, SE-104 05 Stockholm, Sweden
1
Impact of microcystin contaminated irrigation water on the physiology and
mineral content of carrot (Daucus carota) – implications in crop quality
Joana Machado1; Joana Azevedo1; Marisa Freitas2; Edgar Pinto3; Vitor Vasconcelos4; Alexandre
Campos1
The occurrence and proliferation of toxic cyanobacterial blooms are an emergent
environmental concern worldwide. Microcystin-LR (MC-LR), mainly produced by
Microcystis aeruginosa, is the most documented and studied cyanotoxin causing
human health problems [1]. Contaminated waters are commonly used in agriculture
and this may represent a risk to food safety and crop’s quality [2]. The plant root
system is usually more exposed via soil irrigation, and therefore root-vegetables are
more susceptible to contamination. Carrots (Daucus carota) are root-vegetables with
great importance for human nourishment and economy [3]. Our aim was, therefore,
to evaluate the possible negative effects of the use of water containing this toxin in
the physiology and quality of this horticultural crop. For this purpose young carrots
(~ 1 month old) were grown in soil during 1 month in environmental conditions
(room temperature and natural photoperiod). Three groups were performed with
plants being irrigated with non-contaminated water (control group) or with a crude
Microcystis aeruginosa extract containing respectively with 10 and 50 µg/L MC twice a
week. Evaluation of physiological conditions were performed through quantification
of the growth parameters (fresh and dry weight of carrots and leaves) and evaluating
the maximum efficiency of photosystem II, assessed by pulse amplitude modulation
(PAM). The nutritional quality of the carrots was also evaluated by quantifying the
macro (AAS and spectrophotometry) and micro minerals (ICP-MS) content.
The results showed no toxic effects of M. aeruginosa crude extract and MC-LR
toxin at 10 and 50 µg/L concentrations in D. carota, under conditions of sub-chronic
375
exposure at environmental concentrations of the toxin. Moreover, the results showed
Effect of ibuprofen and carbamazepine on cyclooxygenase (COX-2) gill
that both short and prolonged exposure to MC increases photosynthetic efficiency
gene expression, lipids and reproductive hormones in temperature con-
of plants. However, the observed alterations, in the content of macro and micro
ditioned juvenile Solea senegalensis
minerals may have implications on the nutritional value of the carrots and require
González-Mira1, F. Hontoria2, E. Mañanós2, V. Piquer 2, A. Torreblanca1, J.C. Navarro2, and. I. Varó2
further clarification.
Pharmaceutical drugs are dispersed in the environment as a result of human and
veterinary use. The non-steroidal anti-inflammatory drug ibuprofen (IB) and the antiAcknowledgments
Porto University (project IJUP2011_3), the European Regional Development Fund (ERDF) through the
COMPETE - Operational Competitiveness Programme and FCT – Foundation for Science and Technology
(project PEst-C/MAR/LA0015/2013).
convulsive drug carbamazepine (CBZ) are frequently found in aquatic ecosystems at
References
[1] Figueiredo, D.R, Azeiteiro, U.M., Esteves, S.M., Gonçalves, F.J.M., Pereira, M.J. (2004). Microcystin-producin
blooms – a serious global public health issue. Ecotoxicology and Environmental Safety, 59: 151-163
[2] Corbel, S., Mougin, C., Bouaïcha, N. (2014). Cyanobacterial toxins: modes of actions, fate in aquatic and
soil ecosystems, phytotoxicity and bioaccumulation in agricultural crops. Chemosphere, 96: 1-15.
[3] Singh, D. P., Beloy, J., McInerney, J.K., Day, L. (2012). Impact of boron, calcium and genetic factors on
vitamin C, carotenoids, phenolic acids, anthocyanins and antioxidant capacity of carrots (Daucus carota).
Food Chemistry, 132: 1161–1170.
from arachidonic acid (ARA), which can induce alterations in the reproductive
Interdisciplinary Centre of Marine and Environmental Research (CIIMAR/CIMAR), University of Porto;
Polytechnic Institute of Porto. Environmental Health Department. School of Applied Health Technologies.
CISA/Research Center in Environment and Health;
3
REQUIMTE, Department of Chemical Sciences, Laboratory of Bromatology and Hydrology, Faculty of
Pharmacy, University of Porto; 4Department of Biology, Faculty of Sciences, University of Porto
injected intraperitoneally with, 1) IB (10 mg/kg; n=12), 2) CBZ (1 mg/kg; n=12), 3)
1
2
low concentrations. They have the ability to inhibit the synthesis of ciclooxygenases
(COX), reducing the production of prostaglandins (PGs), a group of lipids derived
system. This study investigates the effect of IB and CBZ on COX-2 gene expression
and lipid and fatty acid composition in gills of juvenile Solea senegalensis, acclimated
to two temperatures. Moreover, plasma levels of reproductive hormones, including
androgens (testosterone (T), 11-ketotestosterone (11kt)) and the estrogen estradiol
(E2), were analysed. Soles were acclimatized to 15ºC and 20ºC for 60 days and then,
carrier (sunflower-oil; n=12), 4) non-injected (control; n=12). Two-days after injection,
fish were bled for plasma and sacrificed for dissection of gills. Two-way ANOVA
analysis of COX-2 expression revealed no effect of temperature and treatment, but
the interaction was significant. However, COX-2 expression showed an increasing
trend with temperature. Fatty acid analysis did not show clear segregation patterns
neither by temperature nor by treatment after chemometric analyses. However,
detailed statistical analysis of ARA revealed a significant effect of temperature. At
lower temperature ARA significantly decreased in CBZ-treated fish. Plasma steroids
showed no effect of temperature on E2 levels in females, but in males, androgen
377
levels (T and 11kt) were higher at low temperatures. Drug treatment had no effect in
Influence of temperature on the toxicity of the antibiotic florfenicol to
males, but in females IB and CBZ slightly increased E2 levels at both temperatures,
species representative of freshwater phytoplankton and zooplankton
suggesting potential endocrine disrupting action of these drugs. Work financed by
Maria Alexandra Martins1; Lúcia Guilhermino1
the Spanish Ministry of Science and Innovation (ref-CTM2010-16611), and Generalitat
Valenciana (GV-2014/085-PROMETEO-II).
1Departamento de Biología Funcional y Antropología
Antibiotics are widely used by our Society to prevent and treat several diseases
Física. Universidad de Valencia. Dr. Moliner 50,
Burjassot, 46100 Valencia, Spain.
2Instituto de Acuicultura Torre de la Sal (IATS-CSIC), 12595 Ribera de Cabanes (Castellón), Spain.
in humans and animals, and became environmental contaminants of high concern.
However, their ecotoxicological effects are still poorly understood, especially in
relation to climate changes. To improve the knowledge on this topic, the goal of the
present study was to investigate the influence of temperature rise on the short-term
toxicity of the veterinary antibiotic florfenicol (FLO) to the microalgae Chlorella vulgaris
and cladoceran Daphnia magna, widely used in Ecotoxicology as representative
of freshwater primary producers and primary consumers, respectively. Standard
laboratory toxicity bioassays were carried out with the two species independently at
15ºC, 20ºC and 25ºC, by exposing C. vulgaris and D. magna at different concentrations
of FLO (low ppm range) for 96h. The tested concentrations were selected to allow
the calculation of EC50/LC50 values and do not intend to be ecologically relevant.
The effect criteria were the inhibition of population growth for the microalgae
and mortality for the cladoceran. FLO significantly reduced C. vulgarispopulation
growth with no significant differences among distinct temperatures (EC50s ranging
from 227 to 322 mg/l). In the range of concentrations tested, FLO caused D. magna
mortality, with 96h-LC50s of 351 mg/l at 15ºC, of 319 mg/l at 20ºC and <300 mg/l at
25ºC. Therefore, temperature rise did not influence the toxicity of FLO to C. vulgaris.
However, both the lowest and the highest temperatures tested increased the toxicity
of FLO to D. magna relatively to the toxicity obtained at 20ºC. Overall, these results
show that temperature rise was able to influence the toxicity of FLO to the primary
consumer but not to the primary producer. They also suggest that in real scenarios
379
temperature is able to modulate the effects of FLO on phytoplankton-zooplankton
Assessment of the pharmaceuticals removal by different wastewater
relationships highlighting the need of more research in this topic.
treatments
A.M.P.T. Pereira1; L.J.G. Silva1; C.M. Lino1; L.M. Meisel2; A. Pena1
ICBAS & CIIMAR, University of Porto; ICBAS, Depart Populations Study, Lab of Ecotoxicology,R. Jorge
Viterbo Ferreira, 228, 4050-123 Porto, Portugal; CIIMAR, Research Group of Ecotoxicology, Stress Ecology
and Environmental Health, R dos Bragas, 289, 4050-Porto
1
In recent years, an increased and chronic consumption of several medicines
has been observed all across the world. In Portugal, alprazolam, lorazepam and
zolpidem (anxiolytics and hypnotics), azithromycin and ciprofloxacin (antibiotics),
simvastatin, bezafibrate and gemfibrozil (lipid regulators), and ibuprofen, diclofenac
and paracetamol (non-steroidal anti-inflammatories and analgesics) presented the
highest prescription and consumption (INFARMED, 2011).
After intake, these highly active compounds undergo metabolic transformations,
with subsequent excretion of significant fractions of the unmetabolized or of active
metabolites to raw sewage and wastewater treatment plants (WWTPs). These
facilities are not able to completely remove these contaminants, being considered
the major environmental source (Sousa, 2011).
Our main goal was to evaluate the removal efficiency of 11 of the most consumed
pharmaceuticals, in 15 WWTPs (influents and effluents) with different treatment
processes, during one year (4 sampling campaigns) across Portugal. For identification
and quantification, extraction and cleanup was performed through an Oasis MAX
cartridge and instrumentation analysis was performed by LC-MS-MS (ion trap)
(Sousa, 2011).
WWTPs were not able to completely remove these pharmaceuticals, with
efficiencies ranging from 50.2 to 75.2%. In decreasing order the wastewater
treatments more efficient were activated sludges (AS), biofiltration with ultra violet
(BF/UV), lagoons (LG) with UV, AS/UV and trickling filters (TF) with 75.2, 70.3, 69.2,
64.4 and 50.2% of removal efficiencies, respectively.
381
Despite some differences in the treatments applied, WWTPs exhibited a
Potential of Constructed Wetlands for the removal of antibiotic resistant
comparable performance in their removal, with no statistical significance between
bacteria from livestock wastewater
the different treatments. Nonetheless, it is noticeable a great variation in removal
I. Ribeiro a, b; C.M. Almeida b; F. Santos a, b; A.C. Ferreira a, c; A.P. Mucha b
efficiencies between the different therapeutic groups, as well as within each group,
and no association was establish between the decreased biochemical oxygen
Veterinary pharmaceuticals and their active compounds enter into the aquatic
demand, chemical oxygen demand and total suspended solids in effluents and
systems through discharges of effluents from livestock industry. Among these,
removal percentage.
antibiotics are of main concern, as they can promote the occurrence of antibiotic
resistant bacteria (ARB) among the environmental communities. It has been
proposed that Constructed Wetlands (CW’s) can be used in the reduction of various
Acknowledgments
The authors thank FCT the financial support (project and fellowship PTDC/AAC-AMB/120889/2010,
fellowship granted to L.J.G. Silva SFRH/BPD/62877/2009); Instituto da Água da Região do Norte (IAREN) of
Portugal, for the MS analyses; every entity that provided technical assistance in collecting the wastewater
samples.
References
INFARMED, 2011. Monitorization of the Market. Available at: http://www.infarmed.pt/portal/page/
portal/INFARMED/MONITORIZACAO_DO_MERCADO/OBSERVATORIO/ESTATISTICA_DO_MEDICAMENTO/
EstMed-2011.pdf. Accessed on 04.09.2013, 2011.
Sousa, M.A., Gonçalves, C., Cunha, E., Hajšlová, J., Alpendurada, M.F., 2011. Cleanup strategies and
advantages in the determination of several therapeutic classes of pharmaceuticals in wastewater samples
by SPE-LC-MS/MS. Anal. Bioanal. Chem. 399, 807–22.
LAQV, REQUIMTE, Group of Bromatology, Pharmacognosy and Analytical Sciences, Faculty of Pharmacy,
University of Coimbra, Polo III, Azinhaga de Stª Comba, 3000-548 Coimbra, Portugal;
2INFARMED, I.P. - National Authority of Medicines and Health Products, 1749-004 Lisboa, Portugal
1
pollutants in effluents from livestock industry, including antibiotics. These systems
that simulate some of the removing processes occurring in natural wetlands, present
several advantages, such as low implementation costs and facility of handling. The
present study aims to evaluate the potential of CW’s for the removal of ARB from
livestock wastewater. To simulate a CW system, microcosms (dimensions: 0.4 m x
0.3 m x 0.3 m) were assembled with Phragmites australis and feed with livestock
wastewater doped or not with 100 µg/L of enrofloxacin or 100 µg/L of ceftiofur, two
antibiotics commonly used in livestock industry. Wastewater was treated during oneweek cycle. After the one-week cycle wastewater was removed and replaced by new
wastewater (doped or not), the process being repeated during 18 weeks. Treated
wastewater samples were collected at weeks 1, 2, 4, 8, 14 and 18 to determine the
removal of three groups of ARB (heterotrophic, enterobacteria and enterococci),
which was estimated in plate culture media. Initial livestock wastewater presented
values of ARB that range from 0 to 2 % for heterotrophic bacteria, from 1 to 8 % for
enterobacteria and from 2 to 20 % for enterococci. After each one-week cycle the
removal efficiency was higher than 75% for heterotrophic bacteria, and higher than
383
90% for enterobacteria and enterococci. Therefore, CWs can be a valuable alternative
Cell and Molecular Toxicology
to remove ARB from livestock wastewater, reducing the risk the release of these
effluents might pose to the environment.
Genotoxic effects of rocket propellant’s component on rodents
Anna V. Lovinskaya1; Saule Zh. Kolumbayeva1; Serikbay K. Abilev2; Oksana L. Kolomiets2
Acknowledgements
To European Regional Development Fund through COMPETE - Operational Competitiveness Program and
national funds through FCT, under PEst-C/MAR/LA0015/2013.
Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228,
4050-313 Porto, Portugal
b
CIIMAR/CIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Rua
dos Bragas, 289, 4050-123 Porto, Portugal
c
Faculdade de Ciências Universidade do Porto, Rua do Campo Alegre, s/n, 4169-007 Porto, Portugal
a
The development of the space industry contributes the emergence of new
environmental hazardous factors which may negatively affect the environment and
health. Continuous exploitation of spaceport and rocket launch sites leads to pollution
with propellant components and products of its decomposition and combustion.
There is an acute problem of rocket fuel and its components contamination of
the environment ofKazakhstan, which hosts the Baikonur Cosmodrome. The
results of field work in the fall region of burned-out rocket stages in Russia and
Kazakhstan indicate the presence of the propellant’s component unsymmetrical
dimethylhydrazine (1,1-DMH) and its oxidation products in soil, water and plants
in concentrations exceeding the MAC. Despite the data on the toxic effects of 1,1DMH on the body, its genotoxic effects in somatic and germ cells of mammals
are not well understood. In addition, knowledge of the mechanisms of genotoxic
effects of xenobiotics gives the possibilities for purposeful searches of remedies.
The mutagenic activity and organ-specific genotoxic effects of 1.1-DMH
in BALB/cYwal mice have been studied using cytogenetic and Comet
assay methods. The mechanisms of meiosis infringement in germ cells of
mice have beenstudied using immunocytochemical analysis of synaptonemal
complex (SC). The background rodent species (Citellus pygmaeus Pallas) from 1,1DMH contaminated
areas
have
been
examined
as
well.
The pronounced genotoxic potential of 1,1-DMH in dose- and timedependent effects on animal models (acute and subacute (10 days) exposure) was
385
defined. There was a statistically significant increase of aberrant cells number and
Interference of estrogenic and androgenic inputs in the brown trout lip-
polyploidy
id metabolism – experimental biochemical and molecular studies
level compared
to intact
animals.
1,1-DMH
induced
DNA
single-strand breaks in the cells from visceral organs tissues of mice with
a damage index depending on the dose and duration of the exposure.
1,1-DMH influenced a pronounced genotoxic effect on germ cells of mice. The
statistically significant increase of SC abnormalities of meiosis in 1,1-DMHexposed animals compared to control was determined. That was a result of SC
fragmentation in nuclei, association between SC of sex and autosomal bivalents, and
atypical SC structures. The SC abnormalities in the spermatocytes defined on
38th day after 10 days of subacute 1,1-DMH exposure indicate the risk of long
term preservation of the chromosomal abnormalities occurred in the stem cells.
A high level of structural and genomic mutations in bone marrow cells and DNA
breaks in cells of visceral organs tissues of Citellus pygmaeus from Ulytau district of
Karagandy region exposed to rocket and space activities were identified.
TV Madureira1,2; I Pinheiro1,2; F Malhão1,2; C Lopes1,2; R Urbatzka2; JF Gonçalves1; LFC Castro2,3; E
Rocha1,2
In fish, the lipid-energetic dependent processes – such as reproductive
phenomena – are recognized as being quite susceptible to modulation by
hormones and/or hormone mimics. Nevertheless, it is still much unknown
about the lipid metabolic changes caused by this type of compounds. So,
herein we are interested in studying the effects caused by an estrogen and an
androgen compound in selected plasmatic lipid indices and in the expression
of some genes associated with lipid metabolism, using brown trout as a model.
Juveniles of Salmo trutta f. fario were exposed during 28 days to solvent control – SC
(0.001% of ethanol), 50 µg/L of ethinylestradiol – EE2 and 50 µg/L of testosterone
– T, under semi-static conditions. Then, several lipid parameters (cholesterol,
al-Farabi Kazakh National University, 71 Al-Farabi Av., Almaty, 050040, Republic of Kazakhstan;
N.I. Vavilov Institute of General Genetics Russian Academy of Sciences, 3 Gubkin Str., Moscow, 119333,
Russian Federation
1
2
triglycerides, high-density lipoprotein cholesterol - HDL, low-density lipoprotein
cholesterol- LDL and cholesterol/HDL cholesterol) were evaluated in whole blood or
EDTA plasma. Additionally, the mRNA expression of e.g., apolipoprotein A-I – ApoAI,
fatty acid binding protein 1 – Fabp1, acyl-CoA synthetase long-chain 1 – Acsl1 were
analysed in liver samples, by real-time PCR.
Overall, the major differences in plasmatic levels were observed in the EE2
treatment, where for all lipid parameters there is a clear trend for higher values, in
particular of triglycerides, comparing with the SC animals. The homeostasis of lipid
turnover expression was also affected. In those hyperlipidemic animals, ApoAI and
Fabp1 expressions were drastically reduced, while Acsl1 increased. Those evident
results were not observed in the androgenic exposure group. Facing the observations,
387
we now hypothesize that, among other impacts, the lipidic imbalances caused by
Secale sylvestre: a wild rye potentially useful for aluminum tolerance
hormonal disturbances – which can possibly happen at lower concentrations – may
study
disturb the ovary maturation/reproductive success of this fish.
Elisabete Santos1; Manuela Matos1; Ana Figueiras2; Olinda Pinto-Carnide1; César Benito2
Aluminum (Al) becomes toxic in acid soils that are widespread into the world.
Keywords
Brown trout, in vivo, ethinylestradiol, testosterone, lipid metabolism.
Acknowledgments
FCT (refs SFRH/BPD/97139/2013 and PTDC/CVT/115618/2009).
1Institute of Biomedical Sciences Abel Salazar (ICBAS), U.Porto, Department of Microscopy, Rua de Jorge
Viterbo Ferreira, 228, 4050-313 Porto, Portugal;
2Interdisciplinary Centre of Marine and Environmental Research (CIIMAR), U.Porto, Rua dos Bragas, 289,
4050-123 Porto, Portugal;
3Faculty of Sciences (FCUP), Department of Biology, U.Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal
At low pH, Al3+ ions solubilizes into soil and dramatically inhibits root growth and
function resulting in significant crop yield losses. Among cereals, rye (Secale cereale L.)
has been described as the most tolerant species. Exudation of organic acids from
roots is the most recognized mechanism for Al tolerance. Two main genes that
encode malate (ALMT) and citrate (MATE) aluminum-activated transporter proteins
were characterized in rye.
In this work, Al tolerance of Secale sylvestre was evaluated employing a
hydroponic method with Eryochrome cyanine R staining where the root regrowth
were measured after 24h Al exposure. Thereafter, four histochemical staining
methods were performed in the roots after 24h Al exposure using: morin
(distribution and accumulation of Al), DCF-DA (H2O2 detection), Schiff’s reagent
(lipid peroxidation visualization) and Evans blue solution (cell death screening).
As control, we used barley (Hordeum vulgare) that is sensitive to Al and a sample
of S. sylvestre without Al exposure. The results indicated that S. sylvestre is sensitive
to Al. Moreover, expression studies were done by quantitative RT-PCR for several
candidate genes:ScAlmt1, ScMate1, ScMate2, ScStop (transcription factor involved
in Almt1 and Mate1 expression), ScMdh1 and ScMdh2 (mitochondrial Malate
dehydrogenase), ScCs (mitochondrial Citrate synthase), Fum (Fumarase), ScCSod (Cu
and Zn dependent Superoxide dismutase) and ScMSod (Mn and Fe dependent
Superoxide dismutase). The mRNAs of these genes are more abundant in roots
than in leaves. S. sylvestre seems to be interesting for the study of the mechanisms
389
involved in Al tolerance because is sensitive to this abiotic stress contrary to what
Real time PCR analysis of NMDA receptor expression in zebrafish embry-
happens with the majority of ryes. It is important to compare the expression of these
os exposed to ketamine
candidate genes in this sensitive species with other tolerant ryes.
Cindy Marlene Mesquita Serafim1; Luís Félix1; Luís Antunes2; Manuela Matos3; Ana Maria Coimbra4
Zebrafish (Danio rerio) has emerged as a popular toxicological model.
Center of Agricultural Genomics and Biotechnology (CGBA), University of Trás-os-Montes and Alto Douro
(UTAD), 5001-801 Vila Real, Portugal;
2
Departamento de Genética, Facultad de Biología, Universidad Complutense de Madrid (UCM), 28040 Madrid, Spain
1
Ketamine, a noncompetitive NMDA receptor antagonist, has been widely used
in hospital and as a recreational drug and more recently it has been included
in the group of the emergent contaminants. Some studies suggest that NMDA
receptor antagonist drugs might have neurotoxic direct effects in the developing
brain revealing a relationship between NMDA receptors blockade and
neurodegeneration. This study aimed to analyse the expression of two NMDA
receptor subunits, by RT-PCR, during the first six days of embryonic development,
after exposure to ketamine in early development.
The experimental protocol consisted in two kinds of exposure: a short
exposure (20 min) during blastula, gastrula or segmentation periods to ketamine
concentrations of 0.2, 0.4 or 0.8 mg/mL and a prolonged exposure (21 hours)
to ketamine concentrations of 0.05, 0.07 or 0.09 mg/mL. The expression
in AB-type zebrafish of a consensus sequence of NMDA receptor subunit
genes grind1a and grind1b (NR1) and grind2da and grind2db (NR2) were analysed
by real-time PCR on RNA samples isolated at 4, 8, 24, 72 and 144 hpf embryo/
larvae (n = 50). The housekeeping gene b-actin was included in the analysis as
control.
When ketamine exposure was done during blastula NR1 expression was down
regulated, these subunits gene expression also declined in zebrafish exposed
through 21 hours. By the contrary, an up-regulation was observed when exposed
during segmentation. The NR2 expression was affected in the same way as
391
NR1 expression, but additionally a down regulation was identified on embryos
Short term exposure to ketamine and its oxidative stress in zebrafish
exposed during the gastrula period.
development
The expression of NMDA receptor subunit genes is affected by ketamine
Ana Vidal1; Luís Félix2; Luís Antunes2; Ana M Coimbra2; Sandra M Monteiro2
exposure during early zebrafish development, suggesting that these genes
might be involved in ketamine-induced neurotoxicity during embryogenesis.
Ketamine is a widely used dissociative anesthetic with sedative and analgesic
Additionally, it seems that a late exposure, like during segmentation, might
properties. Due to its psychedelic characteristics, its recreational use is increasing.
interfere with signalling pathways involved in the activation of NMDA receptors,
Ketamine is not completely metabolized in humans and its occurrence in the aquatic
while early and prolonged exposures induce a direct effect in receptors formation.
environment has already been reported. Furthermore, it has been classified as an
emerging contaminant and the biological consequences to aquatic organisms are
1Universidade de Trás-os-Montes e Alto Douro;
2Universidade de Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal; Grupo de Ciências de Animais
de Laboratório, Instituto de Biologia Molecular e Celular (IBMC), Porto, Portugal;
3Universidade de Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal; Instituto de Biotecnologia e
Bioengenharia-Centro de Genómica e Biotecnologia (IBB-CGB UTAD), Vila Real, Portugal;
4Universidade de Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal; Centro de Investigação e de
Tecnologias Agroambientais e Biológicas (CITAB), Vila Real, Portugal
still to be elucidated. In that sense, the knowledge on developmental oxidative
responses is essential. Thus, in this study, we aimed to investigate the short
term effects of ketamine on zebrafish (Danio rerio) as a reliable indicator of early
developmental-induced oxidative stress, once ketamine has been shown to induce
teratogenic effects in this species.
Zebrafish embryos were exposed during 20 minutes to the ketamine
concentrations of 0.2, 0.4 or 0.8 mg/mL, during early blastula, gastrula and
segmentation stages. Six exposure replicates were performed (100 embryos/group).
After exposure,the activities of superoxide dismutase (SOD), catalase (CAT) and
glutathione peroxidase (GPx), glutathione rathio (GSH/GSSG), lipid peroxidation
(LPO), protein carbonyls (PC) and reactive oxygen species (ROS) were evaluated at
8 and 24 hours post-fertilization (hpf ) by colorimetric and fluorometric methods.
The results showed significant differences, at 8 hpf, in the mean levels of SOD and
CAT activities of embryos exposed, at the gastrula phase. Compared to the control
group, ketamine induced a dose-dependent increase on these enzyme activities. No
significant variations were detected on the remaining parameters.
These changes may be attributed to a sensitive window during zebrafish embryo
393
development in which the embryos are more sensitive to ketamine and to a potential
Microbial Ecotoxicology
production of the superoxide ion.
Evidences of salt stress on basidiomycete and zygomycete fungi
Department of Biology and Environment, School of Sciences of Life and Environment, University of Trásos-Montes and Alto Douro (UTAD), Vila Real, Portugal;
2
Department of Biology and Environment, School of Sciences of Life and Environment, University of Trásos-Montes and Alto Douro (UTAD), Vila Real, Portugal | Centre for the Research and Technology of AgroEnvironmental and Biological Sciences, (CITAB), Uni
1
Cátia Venâncio1; Ruth Pereira2; Ana Cristina Freitas3; Teresa Rocha-Santos4; Armando Duarte4;
Isabel Lopes1
Within the context of global climate change, seawater intrusionin lowlying coastal areas isalready happening and it has been projected to become
even more severe. Consequently, increasing concentrationsof salt in terrestrial
ecosystems near coastal areas is expected. Such increased salinity may adversely
affect the growth and development of microbiota that play animportant role in
organic matter breakdown and carbon and nitrogen recycling. The present study,
intended to assessthe effects of increased salinity, due to seawater intrusions,
in the growth of four species of fungi (Pleurotus sajor caju, Phanaerochaete
chrysosporium, Rhizopus oryzae, and Trametes versicolor). For this, two specific
objectives were established: 1) to compare the effects of increased salinity, inthe
growth of the fungi mycelia to assess if NaCl can be used as a surrogate of natural
seawater under the framework of predictive risk assessment and 2) to evaluate
the effects of a long-term exposure to increased salinity in the growth of fungi.
Obtained results revealed that NaClexerts a higher toxicityto these fungi species
than natural seawater. For example, the EC50 obtained for P. chrysosporium and T.
versicolor were31.9 and 16.8 mScm-1 and 37.60 and 34.1 mScm-1, when exposed
to NaCl and natural seawater, respectively. Among the four tested species, P.
chrysosporium showed to be the least sensitive species to increased salinity.The
obtained preliminary results suggest thatNaCl can be used as a safe surrogate to
assess salinization effects caused by seawater intrusionin terrestrial microbiota,
as it seems to constitute a worst-case scenario. Furthermore, these first results
395
suggest that only high levels of seawater intrusion would cause significant
Impacto del cadmio en la actividad nitrificante del bacterioplancton de
adverse effects in white rot fungi and compromise their function in coastal
los canales de Xochimilco (México): estudio en mesocosmos.
terrestrial ecosystems.
M.J. Ferrara-Guerrero1; S. Carballar-Hernández1; J.L. Gama-Flores2; M.E. Huidobro-Salas.2
CESAM/Department of Biology, University of Aveiro;
CESAM/Department of Biology, University of Porto;
3
CESAM/Department of Chemistry,University of Aveiro/ISEIT;
4
CESAM/Department of Chemistry, University of Aveiro
1
En los ecosistemas acuáticos, la comunidad bacteriana es uno de los agentes primarios
2
de la mineralización de la materia orgánica, la regeneración de nutrimentos, fuente de
alimento para los niveles tróficos más altos, son vitales para su buen funcionamiento y
buenos indicadores de estrés del mismo. En este contexto, cualquier factor que afecte
su metabolismo, composición y abundancia es de gran interés. La contaminación por
metales pesados, como el cadmio (Cd), es uno de los factores adversos, frecuentes
y abundantes en la columna de agua y en el sedimento del Lago de Xochimilco
(México). Ante la escasa información del efecto que tiene el Cd, sobre la actividad de las
poblaciones bacterianas que intervienen en los procesos de regeneración del nitrógeno
en este sistema, el objetivo de este estudio fue evaluar (semanalmente) el efecto de
concentraciones (0.0, 0.16 y 0.32 mg/L) del Cd sobre la producción de nitrógeno inorgánico
disuelto (NID), sobre la comunidad bacterioplanctica heterótrofa y nitrificante, y sobre
la tasa de nitrificaciónen el agua de los canales del lago de Xochimilco en condiciones
de mesocosmos, durante un periodo invernal. Se registró el impacto del cadmio con el
tiempo y la concentración tóxica. La biomasa heterótrofa bacteriana osciló entre 1.14
y 5.67 µgC L-1 y se observó una disminución significativa desde la segunda semana,
con una clara recuperación en la cuarta semana. Las bacterias nitrosantes fueron más
susceptibles al tóxico que las nitratantes. La tasa de nitrificación neta disminuyó hasta el
83 %. El medio en los mesocosmos siempre fue fuertemente alcalino, con oxigenación
variable, y con limitación de nitrógeno.
Universidad Autónoma Metropolitana-Xochimilco;
Universidad Nacional Autónoma de México
1
2
397
Nanotoxicology
Effects of silver nanoparticles on different soil exoenzymes activities
Paula Alvarenga1; Márcia Farto1; Clarisse Mourinha1; Daniela Tavares2; Nuno Cruz3; Armando C.
Duarte3; Tito Trindade4; Sónia Rodrigues3; Eduarda Pereira3
Due to their antimicrobial activity, silver nanoparticles (AgNPs) have been widely
used in several commercial products and biological applications, including electronic
products, clothing, cosmetics, and medical applications. The widespread use of AgNPs
will increase their potential inflow into the environment. This may have a detrimental
effect on the activities of microorganisms within the affected environmental
compartment, namely in the soil. The aim of this study was to evaluate the potential
inhibitory effects of the application of AgNPs and of polyvinylpyrrolidone-coated
AgNPs (AgNPs-PVP) on the enzymatic activities of five soil exoenzymes related to
nutrient cycles (β-glucosidase, cellulase, acid phosphatase, protease and urease).
Three types of soils (0-20 cm; air dried; < 2mm), from different regions in Portugal
and with variable soil properties (pH=4.4-6.3; OC=0.80-2.1 %; clay=4.2-23 %), have
been used in this study: Caveira (Cav1), Estarreja (E1), and Penedo Gordo (PG). Water
suspensions (140 mL) of AgNP or AgNP-PVP (20 mg Ag L-1) were applied onto the
surface of pots with 500 g of each soil type (three replicate pots per treatment and
a control of soil without AgNP application were prepared). Pots were kept outdoors
(protected from sunlight, wind and rain) at constant moisture (70% of the water
holding capacity) during 30 days. After 30 days, the soil in each pot was removed
and separated in two halves of 250 g each (top and bottom layer). A sub-sample
of each soil layer was dried and analyzed (in triplicate) for total Ag concentration
by ICP-MS following aqua regiadigestion. The Ag concentration in the soils top
layer was about 11-15 mg kg-1 (median: 13 mg kg-1) while in the bottom soil layer
399
was in the 1.9-7.9 mg kg-1 range (median: 2.6 mg kg-1). There was no evidence for
the influence of the soil properties on Ag retention and distribution in the pots,
but a slightly higher retention of AgNPs-PVP in the soil top layer (median: 14 mg
Ecotoxicological comparison between industrial waste-recovered and
commercial iron oxide nanocatalysts used in ozonation process
T.G. Rauen1,2; V. Z. Baldissarelli1; G. Scaratti1; R. F. P. M. Moreira1
kg-1) was noticed in comparison to non-coated AgNPs (median: 12 mg kg-1). The
soil exoenzymes activities have been analyzed only for the top layer of each pot
Iron oxides recovered from acid mine drainage (AMD) represent a potential
(non-dried soil sub-samples; in triplicate) because the downward mobility of Ag
raw material for the low cost production of goethite with a high degree of purity.
in the soil was relatively limited. For urease, the enzymatic activities were below
The properties of iron oxides enable their use as catalysts in organic pollutant
the quantification limit in all samples (1.6 mmol N-NH4+ g-1 DM h-1). There was not
degradation processes through oxidation methods such as ozonolysis. Thus,
a noticeable effect of the AgNP and of AgNP-PVP on the enzymatic activities of
the toxicity of iron nanoparticles needs to be determined in order to predict
β-glucosidase, cellulase, acid phosphatase and protease. Indeed, no significant
their impact on the environment. In this context, the objective this study was
differences have been observed between the control and the pots of each soil treated
to evaluate the ecotoxicity of goethite and hematite produced from AMD and
with AgNP or AgNP-PVP. In fact, for the soil Ag levels tested here, the influence of soil
compare the results with those obtained for commercial hematite (HCIAL).
origin on the enzymatic activities surpassed that of the application of AgNPs.
The goethite (GTE) was obtained from AMD and then the hematite (H450) was
obtained from calcination of the GTE at 450°C. The materials were characterized
DTCA - Instituto Politécnico de Beja, Rua Pedro Soares S/N, Apartado 6155, 7800-295 Beja, Portugal;
CESAM & CICECO & Dep. de Química, Universidade de Aveiro, 3810-193 Aveiro, Portugal;
3
CESAM & Dep. de Química, Universidade de Aveiro, 3810-193 Aveiro, Portugal;
4
CICECO & Dep. de Química, Universidade de Aveiro, 3810-193 Aveiro, Portugal
1
2
by X-ray diffraction, transmission electron microscopy and determination of the
specific surface area and porosity based on N2adsorption/desorption isotherms.
The Microtox acute toxicity test was performed according to the procedure
described in ISO/DIS 11348-3. The effective concentration at 30 min and 15ºC
(EC50, mg L-1) and the GL values, were quantified using Microtox Omni software,
version 4.0, with a 95% confidence limit. The EC50 values for GTE, H450 and HCIAL
before the ozonation were, respectively, 312.3; 158.2 and 273.6. After 1 h of
ozonation, the corresponding values were 310.0; 162.1 and 283.7, respectively.
These data showed no significant differences in toxicity, indicating some stability
of the nanoparticles, which appear not to be affected by O3. For all samples the GL
values were < 10, indicating low or no toxicity. These results provide an indication
that the iron nanoparticles can be used as catalysts without adverse effects on
401
the environment. However, they need to be considered in combination with an
Quantitative distribution and localization of nanoparticle-derived cop-
investigation of the toxicity on other trophic levels and the characterization of
per in whole cells of Saccharomyces cerevisiae
the samples.
Daniela Correia1; Isabel Sá Correia2; Teresa Pinheiro3
Federal University of Santa Catarina (UFSC – Florianópolis-SC, Brazil);
2
Technological Federal University of Paraná (UTFPR – Francisco Beltrão-PR, Brazil)
1
The aim of this work was to quantitatively assess the cellular uptake of Cu derived
from CuO nanoparticles (CuO-NP) and imaging its distribution in whole cells at
different depths.
The yeast Saccharomyces cerevisiae BY4741 was used as a model. Cells were
incubated in growth minimal medium containing CuO-NP (Nanostructured &
Amorphous Materials, Inc., USA) in concentrations of 15 mgCu/L, 20 mgCu/L
and 40 mgCu/L. Cell growth and viability were assessed. The intracellular and
cell surface Cu distribution was imaged in whole cells using nuclear microscopy
techniques. These techniques, installed at the Van de Graaff accelerator of IST,
can deliver images with micrometre resolution of the cell morphology (scanning
transmission ion microscopy), elemental depth profile (Rutherford backscattering
spectrometry), elemental distributions (particle induced X-ray emission) and
enable quantitative determination of those elements.
The viability of CuO-NP exposed S. cerevisiae ranged from 65% for 15 mgCu/L
to 25% for 40 mgCu/L. The yeast uptake of Cu was proportional to the CuO-NP
concentration in growth medium, i.e., for 15 mgCu/L the total concentration in
whole cells was 2.42 + 1.05 mg/g (dw), for 20 mgCu/ml of 3.39 + 1.26 mg/g (dw)
and for 40 mgCu/L of 7.58 + 2.72 mg/g (dw). Images of different cell depths indicate
that Cu is localized both at the cell surface and inside the cells. A significant fraction
of Cu entered in cells in all concentrations tested. Results highlight the potential of
nuclear microscopy to assess NP-derived metal uptake by cells. In addition, the Cu
distribution patterns and depth profiles can be associated to cell structures.
1Departamento de Bioengenharia, Instituto Superior Técnico, Universidade de Lisboa;
403
2Instituto de Biociências e Bioengenharia, Departamento de Bioengenharia, Instituto Superior Técnico,
Universidade de Lisboa;
3Instituto de Biociências e Bioengenharia, Departamento de Engenharia e Ciências Nucleares, Instituto
Superior Técnico, Universidade de Lisboa
Joint effects of nanoparticles and respective ionic counterparts to
Daphnia magna
S Lopes1; C Pinheiro1; AMVM Soares1; S Loureiro1
The aquatic environment is constantly being exposed to mixture cocktail of
contaminants, including nanoparticles. Despite several studies have already
investigated the adverse effects of many types of NPs to inumerous species of
aquatic organisms, their joint effects is not yet fully understood. In addition
several studies have demonstrated that usually effects are due to both the
particulate and dissolved forms. Therefore, the predictability of their joint effects
is of great importance in environmental risk assessments procedures. Given
the fact that ZnO NPs and Ag NPs are among the most used nanoparticles for
industrial purposes, their co-occurrence in the environment is likely to happen.
Considering this, the present study aimed at assessing the lethal and sublethal
effects of four binary mixtures of two NPs (ZnO NPs and Ag NPs) and respective
ionic counterparts (ZnCl2 and AgNO3) to Daphnia magna. The effects of individual
compounds and their mixtures were studied in lethal and sublethal tests, where
the immobilization and feeding activities were evaluated. Regarding single
compound experiments, both ZnO and ZnCl2 showed similar levels of toxicity
while AgNO3 showed to cause higher toxicity than Ag NPs, for both endpoints
to D. magna. The MIXTOX tool was used to evaluate mixture toxicity. Observed
data was compared with the expected mixture effects predicted by concentration
addition (CA) and independent action (IA) models and potential deviations for
synergistic/antagonistic interactions, dose ratio or level dependency were also
assessed. The general response output for all mixtures of both endpoints was
405
synergism, therefore resulting in a stronger effect than the one expected based
Impact of an organic nanomaterial on soil invertebrates: sds/ddab
on individual chemical toxicity. These results highlight the importance of similar
nanovesicles
studies to understand the combined toxicity in real scenarios.
A. Gavina1; S. Bouguerra2; I. Lopes3; C. Marques3; M. Rasteiro4; F. Antunes4; A. Scheffczyk5; J. Römbke5; N. Graf5; R. Pereira6
1Department of Biology and Centre for Environmental and Marine Studies (CESAM), University of Aveiro,
3810-193 Aveiro, Portugal
The production of nanomaterials (NMs) has been increased due to their vast
potential applications, such as in healthcare and cosmetic products, electronics,
textiles and technology. So far, there is little information about the fate of NMs in
the environment, thus it is necessary to proceed with more research in this area to
understand and characterize the potential effects and risks of NMs to biota. Accordingly,
this work aimed to assess the effects of organic vesicles of sodium dodecyl sulphate/
didodecyl dimethylammonium bromide (SDS/DDAB) in terrestrial invertebrates. For
this purpose were performed several ecotoxicological assays with different species
(Eisenia andrei, Folsomia candida, Echytraeus crypticus and Hypoaspis aculeifer).
A wide range of concentrations of SDS/DDAB were tested, following standard
protocols, and using the standard OECD soil as test substrate (5% of organic matter).
The aqueous suspensions of SDS/DDAB used to spike the soils were characterized by
light scattering techniques for hydrodynamic size of the vesicles, aggregation index,
polydispersity index, zeta potential and surface charge. F. candidareproduction was
affected only when these organisms were exposed to the highest concentrations of
SDS/DDAB (800 and 1000 mg kg-1dw). Earthworms and mites hadn’t their reproduction
inhibited for concentrations up to 1000 mg kg-1dw, however they avoided soil
contaminated with SDS/DDAB. The results gathered in this study allowed us to infer
about the ecotoxicological effects of SDS/DDAB on soil invertebrates, giving rise to
data that could be used in the determination of risk limits for this NM.
Department of Biology, Faculty of Sciences of the University of Porto, Rua do Campo Alegre, s/n, 4169-007
1
407
Porto, Portugal; CIIMAR - Interdisciplinary Centre of Marine and Environmental Research, Rua dos Bragas,
n.289, 4050-123 Porto, Portugal;
2
Laboratory of Water, Energy and Environment (3E), Engineering School of Sfax, University of Sfax,
B.p.w.3038 Sfax-Tunisia;
3
Department of Biology & CESAM, University of Aveiro, Campus de Santiago, 3810-193 Aveiro, Portugal;
4
Department of Chemical Engineering & CIEPQPF & University of Coimbra, 3030-290 Coimbra, Portugal;
5
ECT Oekotoxikologie GmbH. Böttgerstrasse 2-14 D-65439 Flörsheim am Main Germany;
6
Department of Biology, Faculty of Sciences of the University of Porto, Rua do Campo Alegre, s/n, 4169-007
Porto, Portugal; CIIMAR - Interdisciplinary Centre of Marine and Environmental Research, Rua dos Bragas,
n.289, 4050-123 Porto, Portugal
Titanium dioxide nanoparticles exposure under heat shock conditions caused
a partial recovery of respiratory metabolism of Saccharomyces cerevisiae
J Capela-Pires1; R Ferreira2; I Alves-Pereira2
The use of nanomaterials in cosmetics and drugs has grown exponentially due
its reactivity in the living cells. Titanium dioxide nanoparticles (TiO2-NPs) became
the target of toxicological studies because are among the most commonly
used metal nanoparticles. In addition, there are a large number of yeast genes
whose the expression is induced when the cells use glycerol as carbon source
comparatively with cells which use glucose as carbon source. Although the
temperature also affects gene expression, there are no studies involving heat
shock (HS) and NP toxicity. The main purpose of this study was to evaluate
the effects of early exposure of Saccharomyces cerevisiae to 5 µg/mL TiO2-NP
(size<100 nm), with HS in its energetic metabolism. S. cerevisiae UE-ME3, were
grown in YEPG medium (3% glycerol) at 28 ºC.At middle-exponential phase,
2% glucose (YEPGD) and 5 μg/mL TiO2-NPs (size <100 nm) were added to the
culture, and yeasts grown 200 min at 28 ºC or 40 ºC (HS). Biomass was quantified
by dry weight. Post-12,000 g supernatant was used for hexokinase (HK), pyruvate
kinase (PK), lactate dehydrogenase (LDH) and pyruvate decarboxylase (PDC)
determination. The pellet was used for citrate syntase (CS), succinate (SDH) and
NADH dehydrogenase (NDI) determination. The results showed that biomass and
NDI activity of cells grown in YEPGD were higher than those determined in YEPG.
Additionally, was observed that the presence of glucose caused a decrease in the
HK and PK, CS and SDH activity. However, in S. cerevisiae grown in YEPGD with HS
occurred a decrease in the biomass and ALP, CS, SDH, NDI, LDH activity and an
increase of PDC activity. In contrast, simultaneous exposure to TiO2-NPs with HS
409
caused a reverse effect in the CS, SDH and NDI activity, response which suggests
Toxicity of TiO2 nanoparticles on the gills of neotropical fish, Prochilo-
a partial recovery of respiratory metabolism in the presence of glucose.
dus lineatus: bioaccumulation, oxidative stress and oxidative damage
Marisa N Fernandes1; Talita L L Carmo1; Vinicius C Azevedo1; Claudia B R Martinez2; João B Fernandes1
1ICAAM - Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da
Mitra, Ap. 94, 7006-554 Évora, Portugal;
2ICAAM - Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Departamento de Química,
Universidade de Évora, Ap. 94, 7006-554 Évora, Portugal
TiO2 nanoparticles (TiO2-NP) may generate reactive oxidative species (ROS) in
animals’ organs that after interaction with the cellular membranes can cause oxidative
stress. The TiO2-NP bioaccumulation, oxidative stress and oxidative damage were
investigated in the gills ofthe Neotropical fish, Prochilodus lineatus after acute and
subchronic exposure to TiO2-NP.P. lineatus were exposed to0; 0.5; 1.5; 3.0 e 15.0 mgLTiO2-NP during 48 h (acute exposure) and 14 days (subchronic exposure). Thereafter,
1
gills samples were taken; the reactive oxidative species (ROS) production, activity
of antioxidant enzymes (superoxide dismutase – SOD, catalase – CAT, glutathione
peroxidase – GPx) and the levels of non-enzymatic antioxidants (glutathione –
GSH), as well as lipid peroxidation (LPO) were analyzed and the concentration of
titanium in the gill tissue was determined. TiO2-NP bioaccumulation was significantly
increased in the gills of fish after subchronic exposure to 3 and 15 mgL-1 TiO2-NP. ROS
production decreased and the level of GSH increased after acute exposure. However,
after subchronic exposure, ROS production and the activity of SOD decreased in fish
exposed to 15 mgL-1 and GSH increased in fish exposed to 1.5, 3.0 and 15.0 mgL-1. The
activity of CAT and GPx and the levels of LPO did not change. These suggest that GSH
may have an important role in prevent oxidative stress in the gills of P. lineatus exposed
to TiO2-NP. Financial support: FAPESP Proc. 2011/10339-0, 2014/05701-0, CNPq/INCTTA Proc. 573949/2008-5, CNPq Proc. 304279/2011-0.
1Universidade Federal de São Carlos;
2Universidade Estadual de Londrina
411
Itraq-based proteomics analysis of citrate gold nanoparticle exposure
effects in the marine clam Ruditapes phlippinarum.
Moritz Volland1; Amparo Torreblanca2; Julián Blasco1; Miriam Hampel3
Engineered Gold nanoparticles (AuNPs) are introduced into a growing number of
commercial products. Notwithstanding their promising applications, the increased
likelihood of their release into environmental compartments and the resulting exposure
on the species within needs to be met by an increased understanding of their behaviour
and effects. Up to date limited information is available on the ecotoxicological risk for
non-target organisms, with the majority of research focusing on magnified expectable
environmental concentrations. In our laboratory-based study the bivalve Ruditapes
philippinarum was chosen as a model to evaluate differential protein expression of
citrate capped AuNPs (21.5 ± 2.9 nm) at an environmental relevant concentration (0.75
µg L-1). We used a 2nd-generation (iTRAQ-8plex) proteomic approach to identify and
quantify differently expressed proteins in the digestive gland after 1 and 7 days of
exposure in comparison to control organisms.
The iTRAQ results demonstrate that the digestive gland is affected by environmentally
relevant concentrations of citrate capped gold nanoparticles. Over 2500 proteins
involved in a wide range of metabolic pathways and/or physiological processes
were identified. Altered proteins may be useful as biomarkers of environmental
nanoparticle pollution and provide insight about the mode of action of engineered
gold nanoparticles.
Keywords
Citrate capped gold nanoparticles, environmental concentrations, Ruditapes philippinarum, differential
protein expression, iTRAQ-8plex.
1
Departamento de Ecología y Gestión Costera, Instituto de Ciencias Marinas de Andalucía (ICMAN-CSIC),
413
Campus Rio San Pedro. 11510 Puerto Real, Cádiz, España;
2
Departamento de Biología Funcional y Antropología Física, Facultad de Ciencias Biológicas, Universidad
de Valencia. 46100 Burjassot, Valencia, España.;
3
Departamento de Química Física, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz,
Centro Andaluz de Ciencias y Tecnologías Marinas (CACYTMAR), 11510 Puerto Real, Cádiz, España.
Toxicity studies of insecticide-functionalized mesoporous silica
nanoparticles in the milkweed bug Oncopeltus fasciatus
M.A. Ochoa-Zapater1; I.M. Biagoni1; A. Ortiz-Herreros1; F.M. Romero2; A. Ribera2; A. Torreblanca1;
M.D. Garcerá1
In recent years, the application of nanotechnology to the field of agriculture
has resulted in the development of a wide range of products for the protection of
crops described as nanopesticides, which include nanoemulsions, formulations
containing organic nanoparticles and polymer-based formulations. The growing
need for alternative pesticides that delay the onset of resistance phenomena and
prevent adverse effects on non-target organisms are among the main reasons for
the development of these products. Moreover, this new generation of pesticides
can benefit from the properties of nanoformulations as delivery systems for
active ingredients, improving solubility, increasing stability and protecting
it from an early degradation. In this work we have studied the toxic effects of
mesoporous silica nanoparticles (SiO2NPs) functionalized with insecticides
deltamethrin, imidacloprid and fenitrothion in a laboratory reared population
of the milkweed bug, Oncopeltus fasciatus, as a model insect, as we require a
preliminary study for later use of these “nanopesticides” in combating pests in
agriculture. For the synthesis of nanoparticles the methodology described by
Suteewong et al. (2010) was used, while the functionalization was performed
by stirring a mixture of known concentrations of nanoparticles and insecticide
in ethanol, as described in the work from Popat et al. (2012). Acute toxicity
bioassay consisted in the topical application of 1μl of SiO2NPs suspension on the
abdomen of 10 adult males and 10 females (<24 hours old) and mortality rates
were then recorded at 24, 48, 72 and 96 hours after treatment; three replicates
415
were performed for each bioassay. Preliminary results suggest that the use of
Effects of mesoporous silica nanoparticles on food intake and reproduc-
nanoparticles in the formulation of new pesticides can help reduce the amount
tion of Blattella germanica
of active agent required and therefore its impact on other organisms and the
I.M. Biagioni1; M.A. Ochoa-Zapater1; A. Ortiz-Herreros1; A. Ribera2; A. Torreblanca1; M.D. Garcerá1
environment.
Due to the inert composition of SiO2, and the three-dimensional structure that
Acknowledgements
This work has been supported by grant AGL2010-21555 from the Ministerio de Economía y Competitividad.
allows to store different types of substances, mesoporous silica nanoparticles
(SiNPs) are of interest in the synthesis of nanopesticides as potential carriers of
active agents. However, it is necessary to investigate their possible impact on the
Departamento de Biología Funcional y Antropología Física, Universitat de València, Dr. Moliner 50, 46100
Burjassot, Valencia, Spain;
2
Instituto de Ciencia Molecular, Universitat de València, Catedrático José Beltrán, 2. 46980, Paterna,
Valencia, Spain
1
environment and biological systems, because nanoscale materials exhibit different
characteristics and toxicity compared to their counterparts at the macro level. A
previous study of our group on the effects of the ingestion of gold nanoparticles
in Blattella germanica, revealed an increase in food intake and a significant decrease
in the number of nymphs hatched per ootheca, showing that gold nanoparticles
have sublethal toxic effects on this species.
The use of confocal microscope has allowed us to observe that SiNPs labeled
with rhodamine B were able to cross the peritrophic membrane of the intestine
of B. germanica and reach the enteric caeca, indicating that nanopesticides using
silica as a carrier could reach its target within the organism.
In this study we want to investigate the possible effects of SiNPs on food intake
and reproduction of B. germanica. Therefore, we conducted a daily monitoring of
feeding, weight, development and hatching times of the ootheca and the number
of hatched nymphs per ootheca. Preliminary results indicate that SiNPs could affect
the amount of ingested food, the females and their ootheca weight, as well as the
number of hatched nymphs per ootheca.
417
Acknowledgements
This work has been supported by grant AGL2010-21555 from the Ministerio de Econimía y Competitividad.
Departamento de Biología Funcional y Antropología Física, Universitat de València, Dr. Moliner 50, 46100,
Burjassot, Valencia, Spain;
2
Instituto de Ciencia Molecular, Universitat de València, Catedrático José Beltrán 2, 46980, Paterna,
Valencia, Spain
1
Effects of ingested alumina nanoparticles (Al2O3) in the development
and reproduction of Blattella germanica
A. Ortiz-Herreros1; M.A. Ochoa-Zapater1; I.M. Biagoni1; M.D Garcerá1; A. Torreblanca1
There is a need to develop new insecticides to overcome some of the problems
posed by insecticides currently used. Nanotechnology provides materials with new
properties whose effects on living animals are not fully tested. Some of the distinctive
toxic effects of nanoparticles may be useful in pest control (nanopesticides). To
explore this possibility our research group is testing a wide range of functionalised
and non-functionalised nanomaterials. In the present work, the effects of alumnina
Al2O3 (amorphous and in the form of nanoparticles) on parameters related to
reproduction and development of Blatella germanica were determined. Cockroaches
were divided into three experimental groups: a control group, a group treated
with amorphous alumina (0.75mg/g of food) in their feed and a group whose food
contained alumina in form of nanoparticles (30-50 nm). Parameters related to
nutrition (food intake), reproduction (hatching time of egg sacs, number of hatched
nymphs) and survival were measured.
The results allowed us to quantify the toxicity of nanoparticulated and nonparticulated alumina on B. germanica and to evaluate their potential use in pest
control.
Acknowledgements
This work has ben supported by grant AGL2010-21555 from the Ministerio de Economía y Competitividad
Departamento de Biología Funcional y Antropología Física. Universitat de València. Dr. Moliner 50,
Burjassot, 46100 Valencia, Spain
1
419
Nanoparticles toxicity studies on microalgae R. subcapitata
Andreani, T.1; Gavina, A.2; Pereira, R.2; Pinto,V.V.3; Ferreira,M.J.3; Pereira,CM.1
On the recent years, the application of nanoparticles (NPs) on the industry is
widely increasing due to their potential benefits for society and the economy.
Footwear sector is an attractive area for the application of NPs based solutions
in order to develop functional materials for footwear products with control
of bacteria and fungus growth, minimizing the malodors and some foot skin
irritations and infections. Due to increasing production volumes of NPs, there are
considerable concerns from the scientific community about the effects of NPs
to the health human and environment. The objective of this work is to evaluate
the environmental impact of antimicrobial NP using microalgae Raphidocelis
subcapitata. Different solvents were used to suspend the nanoparticles depending
on their solubility. ZnO NP 25 nm were suspended in distilled water or in polyvinyl
alcohol (PVA) 5%, TiO2 NP and CuO NP were dissolved in distilled water, and Cu
NP were dissolved in dimethyl sulfoxide (DMSO) 5%. For the toxicity assay, stock
solutions of NP were prepared in MBL and added to algae into microplate. The
cultures were maintained under photoperiod conditions at a temperature in the
range of 21±2ºC during 72 h, and re-suspended twice a day. The % of algae growth
inhibition was determined by comparing the cell concentration of control with the
exposed cells to NP using an optical microscope. The results showed that ZnO NP
were the most toxic, followed by CuO NP, Cu NP, and TiO2 NP. The toxicity effect
of Zn and Cu NP to algae can be attributed to the solubility of zinc and copper
ions from oxides dissolution. Further studies are being carried out to decrease the
ecotoxicity of these NP. 421
Bioremediation
Acknowledgements
This work was supported by the European Commission through NANOFOOT (Grant Agreement no.
606570).
PGMS removal from aqueous media using anaerobic bacterial communities
from WWTP sludge
Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre 687, 4169-007 Porto, Portugal;
2
Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre 687, 4169-007 Porto, Portugal
& CIIMAR-Interdiciplinary Center of Marine and Environmental Research, Rua dos Bragas 289, 4050-123
Porto, Portugal;
3
CTCP - Centro Tecnológico do Calçado de Portugal, Rua de Fundões - Devesa Velha, 3700-121 São João
da Madeira, Portugal
1
Ana Assunção1; António Candeias2; Maria Clara Costa1
Platinum Group Metals (PGMs) are rare in the Earth´s crust and their use had a considerable
expansion, thus their availability has become more limited, which justifies PGM’s high
commercial value and demand. Therefore, PGMs recovery from effluents contaminated by
these metals is important from economic and environmental points of view.
Recently, several methods for PGMs removal/recovery have been investigated,
including, chemical reductive and electrochemical methods. However, these
processes are economically and environmentally disadvantageous. Thereby, several
biological methods, including the use of bacteria, have been mentioned for that
purpose.Sulfate-reducing bacteria (SRB) have been reported as having the ability to
reduce some metals, including PGMs/Pd(II).
The present work explores the potentialities of bacterial communities enriched
from sludge samples from different systems of municipal wastewater treatment
plants (WWTP), one from a primary Sedimentation Tank (ST) and the other from a
Lagoon System (LS), to remove Pd(II), Pt(IV) and/or Rh(III) from aqueous solutions. For
that purpose the enrichment was done using a modified Postgate medium with and
without sulphate, under anaerobic conditions and just then the obtained consortia
were inoculated in similar media but containing Pd(II), Pt(IV) or Rh(III).
The metals removal results obtained with ST and LS consortia were similar.
Both communities did not show ability to remove Rh(III) (≤11%). The highest Pt(IV)
removal was achieved with the ST inoculum grown in the presence of sulfate (58%).
423
The most encouraging result was obtained for Pd(II) with consortium ST grown
Diversity and densities of bacteria, cyanobacteria and yeasts from waters and
in the presence of sulphate, since the metal (50 mg/L) was completely removed.
sediments of estuarine areas impacted by hydrocarbon in southeast coast
Palladium precipitates were characterized by VP-SEM-EDS. The results suggest that
Ana Julia Fernandes Cardoso de Oliveira1; Aline Bartelochi Pinto2; Vanessa da Costa Andrade2;
Bruna Del Busso Zampieri2; Mirella Massonetto Basilio1
palladium(II) was removed due to the formation of insoluble palladium sulphide. The
finding of a Pd(II)-resistant bacterial community, simply enriched from sludge from
WWTP, with ability for that metal removal can be relevant for the development of
biological technologies aiming PGMs recovery.
The coastal zone is subject to numerous pressures and impacts caused by human
activities, including those of oil exploration that launching in estuaries and other
marine ecosystems various pollutants including polyaromatic hydrocarbons. The
sediment are the most affected by hydrocarbons substrate. Some indigenous
Centro de Ciências do Mar (CCMAR); Faculdade de Ciências e de Tecnologia, Universidade do Algarve,
Campus de Gambelas, 8005-139 Faro, Portugal; 2HERCULES Laboratory, Escola de Ciências e Tecnologia,
Universidade de Évora, Largo Marques de Marialva 8, 7000-809, Portugal
1
microorganisms have affinity for hydrocarbons and are capable of degrading these
compounds enable them to be used as bioremediators. In order to characterize and
quantify microorganisms present in estuarine sediments the densities and divesity
of cyanobacteria, heterotrophic bacteria and yeasts were evaluated in two Estuaries
of São Paulo State (Brazil), Santos Estuary (contaminated by hydrocarbons) and
Estuarine System of Cananéia (preserved area). The densities of microorganisms
were determined by direct counting and the identification was made by morphology
of strains and traditional biochemical tests. Densities of cyanobacteria and
heterotrophic bacteria were higher in Santos sediments than in the Cananéia. On the
other hand, the densities of yeasts were higher in Cananéia. While in the water and
sediments samples of Santos predominated Pseudomonas sp, Mycobacterium sp
and Aeromonas sp, in samples of Cananéia, the predominant genera
were Vibrio sp, Bacillus sp and Staphylococcus sp. Candida sp was dominant in waters
and sediments of Santos estuary. Very important was the presence of Exophialia
dermatitidis, a pathogenic yeast, both in the water and in the sediments of Santos
Estuary. Regarding cyanobacteria, in both estuaries, the number of strains isolated in
the sediment was higher than strains isolated from water samples although in Santos
sediment the number of isolates was almost 3 times higher than that isolated from
425
Cananéia. Most of cyanobacteria strains isolated were identified as Synechococcus sp
Resistance and bioremoval of paracetamol by a sulphate-reducing bac-
and Prochlorococcus. The numerical dominance and the specific composition of
teria community
the isolated heterotrophic bacteria and especially of cyanobacteria, in estuarine
Tânia Cristina da Luz Palma1; Maria Clara Costa1
sediment samples with higher concentrations of hydrocarbons indicate that species
of these microorganisms have potential to be used in bioremediation.
Paracetamol also known as acetaminophenis the most widely used analgesic
and antipyretic all over the world, easily accumulated in the aquatic environment.
It has been detected in surface waters, wastewater, and drinking water throughout
UNESP - Campus do Litoral Paulista; 2UNESP - Instituto de Biociências de Rio Claro
1
the world, thus is a recalcitrant molecule occurring in wastewaters that cannot
usually be removed in WWTP by conventional treatment trains. It has been found
in concentrations up to 6 mg/L in European WWTP effluents. In USA waters it was
determined to occur at a maximum concentration of 10 µg/mL. Its presence in treated
wastewaters causes recognized negative impacts in the receiving water bodies
and human health. Therefore, the search for new and effective ways to degrade
or remove this compound is extremely important. Although (photo)chemical
degradation has been extensively investigated, microbial degradation is largely
unknown. Microorganisms and particularly bacteria have already demonstrated to
play an important role on the biodegradation of organic compounds. The present
investigation is focused on the search of bacterial biodegrading communities, taking
into account the well-known advantages of consortia over pure cultures.
In this study we used an anaerobic bacterial community of Sulphate-Reducing
Bacteria (SRB) enriched from an environmental sample, to test its resistance and
ability to degrade paracetamol. The results show that the SRB consortium is resistant
to concentrations of paracetamol from 5 µg/ml to 100 µg/L. The performance and
growth of this consortium is even better and faster in the presence of this drug
(100 µg/mL) than in the positive control, where no drug was added to the nutrient
medium. Sulphate reduction was complete in the presence of 5 to 100 µg/mL of
427
paracetamol, whereas the removal of paracetamol was higher than 50% and 65% for
Effect of 24-epibrassinolide on ros content, antioxidant enzymes and Ni
both concentrations after 14 and 22 days of incubation, respectively. Compared to the
accumulation in Solanum nigrum L. under Ni stress
negative control the results suggest the occurrence of paracetamol biodegradation.
C. Soares1; A. de Sousa1; A. Pinto1; M. Azenha2; J.Teixeira1; F. Fidalgo1
Nickel (Ni) is one of the major world pollutants, leading to many physiological
Centro de Ciências do Mar (CCMAR); Faculdade de Ciências e de Tecnologia, Universidade do Algarve,
Campus de Gambelas, 8005-139 Faro, Portugal
1
disturbances in plants. Solanum nigrum L. is capable to survive in polluted
sites, tolerating high levels of HM in the soil. Brassinosteroids (BRs) are steroid
phytohormones known to induce tolerance to several abiotic stresses in plants.
In the present work, the effect of exogenous application of 24-epibrassinolide
(EBL) on enzymatic antioxidant system (superoxide dismutase, SOD; catalase, CAT;
ascorbate peroxidase, APX) and on ROS (H2O2 and O2.-) levels was investigated in S.
nigrum plants exposed to high levels of Ni (100 μM). In parallel, Ni accumulation in
shoots and roots of plants was also estimated.
Shoots of Ni- and EBL/Ni-treated plants exhibited lower O2.- and H2O2 levels than
the control, a rise of SOD and APX activities and a decrease in CAT activity, even
though this pattern was more evident in plants exposed to Ni alone. Regarding roots,
the Ni treatment caused a decline in H2O2 and an increase of O2.-, both in presence
or absence of EBL; SOD and CAT activities in plants exposed to Ni alone were higher,
while APX activity did not show significant changes with respect to control plants;
EBL pretreatment resulted in a decrease of both CAT and APX activities, but SOD
activity was positively affected. Ni-treated plants showed an increase in Ni content,
with a higher accumulation in roots than in shoots, regardless of being or not
pretreated with EBL.
The exposure of S. nigrum to Ni 100 μM did not induce a severe oxidative damage,
and consequently, the anti-stress properties of EBL against Ni toxicity weren’t so
evident. However, previous results(1) and data of the present study, strongly support
429
the evidence that the EBL pretreatment ameliorated Ni phytotoxicity, possibly by
Studies on the biodegradability of two environmentally relevant aliphatic
limiting Ni root uptake and increasing non-enzymatic antioxidants content .
organofluorines
(1)
Diogo A. M. Alexandrino1; Luís M. Pinto2; Inês Ribeiro2; Rui. S. Oliveira3; Maria F. Carvalho1
BioISI - Biosystems & Integrative Sciences Institute, Departamento de Biologia, Faculdade de Ciências,
Universidade do Porto, Rua Campo Alegre s/n, 4169-007 Porto, Portugal; 2CIQ-UP, Departamento de
Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, Rua Campo Alegre 687, 4169-007
Porto, Portugal
1
Soares, C., Teixeira, J. and Fidalgo, F. 2014. Foliar application of 24-epibrassinolide in Solanum nigrum L.
exposed to high levels of Ni. IJUP’14 – 7th Meeting of Young Researchers of University of Porto. Porto,
Portugal.
(1)
This work was supported by Project ref. GEHEE33K - Acordo de Cooperação Internacional I&D USP/U.Porto.
Organofluorines are an important group of synthetic chemicals with a
widespread use in modern societies. Due to their useful properties, these
compounds are used in a multitude of applications and, as a result, fluorinated
compounds have been increasingly found as environmental pollutants. Trifluoroacetate (TFA) is a highly recalcitrant, water soluble and extremely stable
fluorinated aliphatic carboxylic compound. It is used in various industrial applications
and is also a final product from the atmospheric degradation of hydrofluorocarbons
and hydrochlorofluorocarbons. Fluoroacetate (FA) is a compound naturally
produced by certain plants and is mainly used as a pesticide, being also a metabolite
of many other fluorinated pesticides, industrial chemicals, anticancer agents and
narcotic analgesics. Both compuounds have been detected in several environmental
compartments, with special focus in aquatic environments. FA is known to be degraded
by several microbial species while biodegradation of TFA has rarely been reported.
Together with the lack of biological mechanisms to biodegrade TFA, there is also a
gap of knowledge on the biodegradability of FA and TFA when present as a mixture.
In this context, this work aimed to investigate the biodegradability FA and TFA, by testing
these compounds: (i) as sole carbon sources; (ii) in cometabolism with acetate (only for the
case of TFA) and (iii) as a mixture of the two carbon sources. The experiments were carried
out in batch mode in aerobic conditions, using microbial inocula from different origins.
Biodegradation of TFA under anaerobic conditions was also investigated. The results
431
obtained so far indicate that FA is degraded by almost all the tested microbial cultures, with
Phytofiltration of uranium-contaminated waters in laboratory conditions
9 microbial isolates derived from these cultures being capable to completely defluorinate
João Pratas1; Paulo J. C. Favas2
this compound. TFA was shown to be highly recalcitrant under all the conditions tested.
When present as a mixture, biodegradation of FA was not affected by the presence of TFA,
while TFA remains recalcitrant under these conditions.
The objectives of the present study are: (1) to develop a phytofiltration prototype system;
(2) to test the potential of some native aquatic plant species for the decontamination of
U-contaminated waters. The species Callitriche stagnalis Scop., Potamogeton natans L.
and Potamogeton pectinatus L. of the native plant community of the uraniferous region
CIIMAR – Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua dos
Bragas 289 – 4050-123 Porto, Portugal
2
Department of Environmental Health, School of Allied Health Sciences, Polytechnic Institute of Porto,
Rua Valente Perfeito 322 – 4400-330 Vila Nova de Gaia, Portugal
3
Research Centre on Environment and Health, School of Allied Health Sciences, Polytechnic Institute of
Porto, Rua Valente Perfeito 322 – 4400-330 Vila Nova de Gaia, Portugal
4
Centre for Functional Ecology, Department of Life Sciences, University of Coimbra, Calçada Martim de
Freitas – 3000-456 Coimbra, Portugal
1
of Beiras (Central Portugal) were selected because they are autochthonous and they show
high accumulation levels and/or high biomass production. Fluorometry was adopted for
the determination of the U content in the water and plant samples using a“Fluorat-02–2M
analyzer” (Lumex, Russia). The installed prototype consists of a closed circuit of channels.
The system was initially contaminated with 500 µg/L of U as uranyl. The phytoremediation
test lasted 15 days, and plant and water samples were collected at the beginning of the
test and after 1, 2, 7, and 15 days of continuous water circulation. The performance of this
system was very effective. The U concentration in the water dropped to 220 µg/L in 24
hours and after two weeks it had decreased to 72.3 µg/L, thus representing an efficiency
of 85.5%. The U concentration in C. stagnalis increased from 0.98 to 1567 mg/kg, in P.
natans increased from 3.46 to 271 mg/kg and in P. pectinatus increased from 2.63 to
1588 mg/kg. The selected plant species are a promising choice for the remediation of
low to medium U contaminated waters. However, to assess the real phytotechnological
potential of the proposed system, and before an in situ application, laboratory tests are
needed to determine the toxicity level, biomass productivity and possible enhancement
strategies for U uptake. The issue of the final disposal of the residues generated also has
to be addressed.
1University of Coimbra;
2University of Trás-os-Montes e Alto Douro
433
Distribution of Pollutants in the Biosphere
Metal(loid) allocation in the soil-plant system of a Pinus halepensis population growing on semiarid mine tailings
I. Párraga-Aguado1; M.N. González-Alcaraz1,2; M.C. Tercero-Gómez1; J. Álvarez-Rogel1; H.M. Conesa1
Aleppo pine (Pinus halepensis Mill.) is a widely used species for restoring
degraded semiarid areas. Nevertheless, its use for the restoration of metal(loid)
polluted soils has not been thoroughly investigated. The goal of this study was to
evaluate the metal(loid) accumulation and nutrient internal cycling in P. halepensis
trees growing at a mine tailings disposal site in a semiarid area of Southeast Spain.
For this purpose, the metal(loid) (As, Cd, Cu, Fe, Mn, Ni, Pb, Sb and Zn) and nutrient
(N, P, K, S, Ca, Mg) concentrations in the foliage (young or current-year needles and
one-year-old needles), woody stems and fresh leaf litter of pine trees were analysed.
In addition, tailings soil properties below pine litter and bare soil were characterised.
Young (current-year) needles showed lower accumulation of metal(loid)s and higher
nutrient concentrations (P, K) than older needles. Pine trees showed significant
accumulation of Cd, Cu, Pb and Sb in woody tissues (e.g. 4.93 mg kg-1 Cu). By other
side, high concentrations of As, Cd, Sb, Pb and Zn occurred at leaf litter (e.g. 87 mg
kg-1 Pb). Although it can be hypothesized that metal(loid)s will return to the soil in
soluble forms via decomposition, in this study water-extractable concentrations in
the soil below the litter layer did not reveal greater release of metal(loid)s compared
to the bare soil. This is probably due to the recalcitrant nature of pine needles and
the slow mineralisation typical of semiarid environments. However, these results
should be contrasted with long-term studies in order to assess the risk of metal(loid)
s leaching and transfer into the food chain. Funding: Fundación Séneca of Murcia
435
(15296/PI/10) and MINECO of Spain and FEDER (CTM2011-23958). Dr. H.M. Conesa
Levels and spatial distribution of organochlorinated compounds and poly-
thanks MINECO and UPCT for “Ramón y Cajal´´ programme (RYC-2010-05665). Dr.
cyclic aromatic hydrocarbons in fishes from a mediterranean coastal lagoon
M.N. González-Alcaraz holds a postdoctoral grant from Fundación Ramón Areces.
Víctor M. León1; Rubén Moreno-González1; Concepción Martínez-Gómez1; Inés García1; Carlota
Ceruso1; Fulgencio Martínez1; Víctor García1; Juan A. Campillo1
Universidad Politécnica de Cartagena, Departamento de Ciencia y Tecnología Agraria, Escuela Técnica
Superior de Ingeniería Agronómica. Paseo Alfonso XIII, Nº 48, 30202, Cartagena, Spain;
2
(current address) VU University of Amsterdam, Department of Ecological Science, Faculty of Earth and Life
Sciences. De Booleaan 1085 1081, HV Amsterdam, The Netherlands.
1
The seasonal input and distribution of organic contaminants in the Mar Menor
lagoon have been recently characterized in seawater and sediments, showing the
significant impact of flash flood events. Mar Menor is a hypersaline (42-47 psu) coastal
lagoon located in the Cartagena Field area at the South East of Spain subjected
mainly to intensive agriculture, seasonal tourism and recreational activities. The
bioaccumulation of polycyclic aromatic hydrocarbons (PAHs), polychlorinated
biphenyls (PCBs) and organochlorinated pesticides (OCPs) has been also determined
in bivalves, but up to date no information was available about their accumulation
in fishes. In this study the concentrations of PAHs, PCBs and OCPs in golden grey
mullet (Liza aurata) and red mullet (Mullus barbatus) muscle were determined.
Their spatial distribution throughout the Mar Menor lagoon was characterized
considering 9 sampling areas and two different sampling periods, spring and
autumn in 2010 for golden grey mullet and only one (autumn) for red mullet. PAHs
and organochlorinated pollutants were extracted using specific Soxhlet procedures.
After cleanup and purification steps the final extracts were analyzed by HPLC for
PAHs, and by GC-MS for PCBs and OCPs. The higher concentrations of all pollutants
were found in fish sampled close to main urban and port areas of this lagoon and
areas under the influence of El Albujón watercourse. PAHs concentrations (sum of 14
congeners) in both species varied between 1 and 20.7 µg/kg w.w., being pyrene and
phenanthrene the most abundant ones. p,p’DDE showed higher concentrations than
PAHs in golden grey mullet (concentration range: 0.2-32.7 µg/kg w.w.), specially close
437
to the main navigation channel between this lagoon and the Mediterranean Sea.
Acknowledgement
This work has been supported by the Spanish Inter-Ministerial Science and Technology Commission
through ‘DECOMAR’ project (CICYT, CTM2008-01832) and by Seneca Foundation (Region of Murcia, Spain)
through ‘BIOMARO’ project (15398/PI/10). Rubén Moreno-González wishes to thank the Spanish Ministry
of Science and Innovation for the FPI grant (BES 2009-014713).
Instituto Español de Oceanografía, Centro Oceanográfico de Murcia
1
Vertical distribution of PAHs in marine sediments (particulate fraction
and interstitial water) from eight iberian mediterranean areas
Víctor M. León1; Rubén Moreno-González1; Carlota Ceruso1; Inés García1; Concepción
Martínez-Gómez1; Juan A. Campillo1
Polycyclic aromatic hydrocarbons (PAHs) have been determined in sediment
cores from eight Iberian Mediterranean coastal areas (Barcelona, Tarragona, Ebro
Delta, Valencia, Castellón, Cartagena, Almería y Málaga), considering both interstitial
water and solid fraction. Three sediment cores were taken in three different sampling
sites per area by using a box corer. Cores were cut into 1-cm-thick sections between
0 and 18-cm depth, interstitial water was obtained by centrifugation and samples
from every site were pooled. Fourteen polycyclic aromatic hydrocarbons (fluorene,
phenanthrene, anthracene, fluoranthene, pyrene, benz[a]anthracene, crysene,
benzo[e]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene,
benzo[g,h,i]perylene, dibenzo[a,h]anthracene and indeno[1,2,3-c,d]pyrene) were
determined in interstitial water by stir bar sorptive extraction coupled to GC/MS and
solid fraction by Soxhlet extraction and HPLC with fluorescence detection. As result
of PAHs hydrofobicity, their concentrations were several magnitude order higher in
sediment (solid phase) than interstitial water. Total PAH concentration in sediment
varied from 1 to 1321 µg·kg-1 d.w and from 4.9 to 274 ng·L-1 in interstitial water.
Vertical PAHs distributions in sediments were homogeneous in some areas, however,
sharp concentration decreases were observed at deeper layers from some sampling
sites. However, their maximum concentrations in interstitial water were found mainly
in upper or in deeper layers, depending on the specific characteristics of the area and
sediments.
439
Acknowledgements
This work was supported by the Spanish Ministry of Environment (2-ESMARME and 2-2-ESMARME
projects), the Spanish Inter-Ministerial Science and Technology Commission through the ‘IMPACTA’ (CICYT,
CTM2013-48194-C3-1-R) project and the European Union through the European Regional Development
Fund (ERDF).
Instituto Español de Oceanografía, Centro Oceanográfico de Murcia
In-situ phytoextraction os nickel by a native population of Alyssum serpylofolium on ultramafic sites (Portugal): prospects for agromining
Igor Morais1; Joana Campos1; Paulo J. C. Favas2; João Pratas1; Fernando Pita1; M. N. V. Prasad3
The present study had the following objectives: (1) to verify variability in Ni
1
accumulation by Alyssum serpyllifolium ssp. lusitanicum at different locations of
ultramafic areas of Portugal; (2) to investigate the relationship between the metal
concentration in the soil (total and bioavailable fraction) and the concentration in the
plant; and (3) to preliminarily assess the potential for this species in agromining and
phytomining Ni as bio-ore. This species is endemic to serpentine soils of the Bragança
and Morais massifs and is a Ni hyperaccumulator containing more than 7,000 mg kg–1
(dry weight) of this element. Under natural conditions, the Ni concentrations found in the
plants in both ultramafic massifs, expressed as dry weight, ranged from 0.15% to 0.66%
Ni, with a mean of 0.41% Ni (0.35% mean in the Bragança massif and 0.46% mean in
the Morais massif). The results show that considering only the aerial parts of the plant,
the biomass, varies between 7.04–9.37 tonnes per hectare, containing a range of 0.12–
0.70% Ni, allowing a withdrawal of between 12.2–44.0 kg Ni ha–1 per crop under natural
conditions. The amount of Ni extracted by the plant is similar in both ultramafic areas,
having a mean value of approximately $US 500 per ha considering the August 2014 Ni
prices.A relevant feature of the present work is its focus on the Ni content of the entire
aerial parts of the plants, whereas many previous reports have quoted analyses of leaves
only, this generally being the most Ni-rich material. From the point of view of agromining
(or phytomining) it is valuable to have realistic data on the plant parts that might actually
be harvested.The preliminary results of this study indicate that some of the land in these
ultramafic areas are potential producers of Ni from this endemic species.
University of Coimbra; 2University of Trás-os-Montes e Alto Douro; 3University of Hyderabad
1
441
Uranium uptake by native terrestrial and aquatic plants in an uranium
mine (central Portugal)
Paulo J. C. Favas1; João Pratas2
The main objective of this study was investigate the uranium (U) biogeochemistry
in the soil–plant and water–plant systems in an abandoned U mine of Central Portugal.
Samples of soil, water, terrestrial plants and aquatic plants were collected. It was
intended to obtain a global view of the response of the vegetation to the accumulation
of U. This implied sample collection in the biogeochemical background and within the
anomaly. As a result 82 different species were identified and sampled. These species
belong to 32 families, five of which are aquatic. Uranium soil contamination in the mine
area ranges from 8 to 560 mg/kg. In the most abundant families of terrestrial plants, the
results show that the Asteraceae and Ericaceae families have the highest concentrations.
For the Asteraceae, an average of 4.91 mg/kg and a maximum of 13.12 mg/kg was
found in Helichrysum stoechas and an average of 4.07 mg/kg and a maximum of 10.52
mg/kg was recorded in Hypochaeris radicata. In Erica umbellata an average of 1.70
mg/kg and a maximum of 7.50 mg/kg maximum were obtained. Even though the
concentrations obtained in this later species are not high it is particularly interesting
because it has a high bio-productivity. Although the U concentrations in waters did not
surpass 13 µg/L we have verified that in the vicinities of the mine drainage area, the U
concentrations in aquatic plants are higher than in the surrounding water bodies. Four
these species proved to be U accumulators with maximum concentrations: Callitriche
stagnallis (55.53 mg/kg), Lemna minor (52.98 mg/kg),Riccia fluitans (50.59 mg/kg), and
Lythrum portula (32.93 mg/kg).The plant survey revealed that some of the native plant
species are well adapted to U contamination in soils and water.
University of Trás-os-Montes e Alto Douro; 2University of Coimbra
1
443
Environmental Chemistry And Toxicology
Repercusión de los plaguicidas utilizados en horticultura protegida sobre el perfil tiroideo y la colinesterasa plasmática (Salto-Uruguay).
María Alexandra Bozzo de Brum1; Raquel de Souza Viera2; Oscar Irabuena3; María Emilia Cassanello4; Elena Esteves5; Andrea Texo6; Natalia Dalmao7; Daniela Fernandez7; Omar Sena8; Mónica
Cadenazzi9
La exposición a plaguicidas en los trabajadores de invernaderos es considerada
de alto riesgo para la salud humana. Las condiciones de altas temperaturas y
humedad pueden incrementar la probabilidad y gravedad del daño. El objetivo de
esta investigación fue: evaluar el estado de salud de las personas expuestas laboral y
ambientalmente a plaguicidas en cultivos hortícolas protegidos de tomate y morrón.
Se incluyó una población de 114 personas expuestas y 96 personas control. Se analizó:
colinesterasa plasmática en sangre, hemograma, perfil hepático y renal y el funcional
tiroideo (TSH, T3 libre y T4 libre) en ambas poblaciones. La disminución promedio de
la enzima colinesterasa de su nivel basal en la población expuesta fue de un 34%,
67% y 48% en el 13% de los aplicadores, 20% de las amas de casa habitantes del
predio y 10% de los trabajadores no aplicadores respectivamente lo que refleja una
absorción de plaguicidas organofosforados y carbamatos. La Ordenanza Ministerial
145/ 2009 del Ministerio de Salud Pública determina que un descenso del 30% de
la colinesterasa plasmática se considera significativo e implica el retiro inmediato
del trabajador de la exposición y evaluación médica. El 3% de la población control
y el 35% de la población expuesta presentaron valores Transaminasa GlutámicoOxalacética (enzima del hígado), por encima de 50 unidades respectivamente (valores
de referencia: entre 10 y 40 unidades) lo que podría estar asociado al herbicida
Paraquat utilizado por el 63% de los productores. No se evidenciaron diferencias
445
significativas en el funcional tiroideo entre la población expuesta y la control lo que
Estudio de los efectos del azufre y del caolín sobre Drosophila suzukii.
podría deberse al tamaño de la muestra. No se halló un patrón definido entre la
G. Albendín1; A. Delfín1; F. Soto-Mancera1; J. Arellano1; M.I. Arufe1; J.M. Molina2; S. Pérez2
concentración de hormonas tiroideas en sangre con la edad y los años de exposición.
Esta investigación muestra que la población expuesta laboral y ambientalmente a
Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) es una plaga
plaguicidas presentó alteraciones enzimáticas.
invasora originaria del sudeste asiático que amenaza seriamente a varios
Universidad de la República-Regional Norte-Facultad de Agronomía, C.P. 50000;
2
Universidad de la República- Facultad de Medicina, C.P. 11800;
3
Universidad de la República- Regional Norte, Laboratorio de Inmunología, C.P 50000;
4
Universidad de la República-Facultad de Agronomía, Estación Experimental San Antonio (E.E.F.A.S),
C.P.50009;
5
Hospital Regional Salto, C.P.50000;
6
Universidad de la República-Regional Norte, Biología Humana, C.P 50000;
7
Universidad de la República, Escuela Universitaria de Tecnología Médica (EUTM), C.P. 60000;
8
Universidad de la República-Regional Norte, Área de Laboratorios, C.P 50000;
9
Universidad de la República-Facultad de Agronomía, Estación Experimental Mario Cassinoni (E.E.M.A.C.),
C.P. 60000.
cultivos importantes en Europa, donde fue detectada por primera vez en 2008.
1
Esta especie ataca fruta sana y madura, convirtiéndola en grave amenaza
para numerosas especies cultivadas como los frutos rojos. En la actualidad,
la principal estrategia de control para esta especie se basa en la utilización
de insecticidas dirigidos a los adultos. Sin embargo, la efectividad de estos
tratamientos y su efecto en la fauna útil no ha sido suficientemente probada
y un uso repetitivo podría provocar la aparición de poblaciones resistentes,
por lo que la selección de materias activas alternativas resulta imperativa. Las
propiedades insecticidas del azufre han hecho que haya un interés creciente
en incluir este producto en los Programas de Manejo Integrado de plagas
como alternativa al uso de insecticidas orgánicos de síntesis. Por otra parte,
el caolín ha mostrado ser efectivo como barrera de puesta para algunas
plagas importantes como Batrocera olea o Prays oleae en el cultivo del olivo.
Este trabajo analiza el potencial uso de azufre y caolín para el control de D.
suzukii en frutos rojos. Los ensayos se llevaron a cabo en laboratorio utilizando
arándanos de variedades comerciales y adultos de D. suzukii procedentes de
una colonia establecida en cautividad en el Laboratorio de Entomología del
Centro IFAPA Las Torres-Tomejil. Para cada producto se llevaron a cabo dos
tipos de ensayo, ensayo de no elección y ensayo de elección, y se estudió
el número de huevos puestos por fruto, el número de adultos emergidos,
mortalidad y potencial efecto sobre la actividad acetilcolinesterasa. En relación
447
con este biomarcador, previamente se realizó la caracterización de las enzimas
Biliary PAH metabolites in european eel (Anguilla anguilla) from Mar
colinesterasas presentes en esta especie mediante el uso de diversos sustratos
Menor Lagoon (SE Spain)
e inhibidores específicos y la determinación de las constates cinéticas Vmax y
Ángel Gea-Pacheco1; Rubén Moreno-González1; Concepción Martínez-Gómez1; Dick Vethaak2;
Victor M. León1
K m. Aunque son necesarios ensayos que confirmen su efectividad en campo,
los resultados obtenidos en este trabajo ponen de manifiesto el potencial del
azufre micronizado para su uso como barrera de puesta y recubrimiento de
fruto contra D. suzukii, ofreciendo una alternativa a los insecticidas orgánicos
de síntesis compatible con el manejo ecológico del cultivo.
Laboratorio de Toxicología. Facultad de Ciencias del Mar y Ambientales. Universidad de Cádiz. CASEM,
Polígono Río San Pedro 11510. Puerto Real, Cádiz (España).
2
Laboratorio de Entomología. IFAPA Centro Las Torres-Tomejil. Junta de Andalucía. Ctra. Sevilla-Cazalla
km12.2, 41200 Alcalá del Río, Sevilla (España).
1
Polycyclic aromatic hydrocarbons (PAHs) areubiquitous organic contaminants
present in marine sediments as a consequence of their continuous input from
either land- or marine-based sources. Fishes have a higher capacity to metabolize
and excrete PAHs than invertebrates such as mollusks and consequently the
concentration of PAH metabolites in the bile fluid of fishcan be used as an indirect
indicator of exposure to PAH contamination. In this study the concentration and
distribution of major PAHs metabolites in European eel (Anguilla anguilla) bile from
the hypersaline Mar Menor coastal lagoon (SE Spain) were characterized for the
first time. Eels of two different weight classes (<350 g and >350 g) were sampled
from the northern and southern part of the lagoon using traditional fishing
methods by local fishermen in spring 2014 and winter 2015. Bile samples were
treated individually and maintained at -20ºC until analysis. 10 µL of bile sample was
diluted with water for liquid chromatography, incubated with β-glucuronidase/
arylsulfatase for 2 hours at 37ºC, then the reaction was stopped with cold methanol
and the sample centrifuged. The concentrations of PAHs metabolites (phenanthrol
and pyrenol) in the supernatant were directly analysed by liquid chromatography
with fluorescence detection using a standard solution for external calibration.
Phenanthrol and pyrenol were found in all samples, with pyrenol being always
the predominant one. The metabolite concentrations were higher in specimens
sampled in spring 2014 than in winter 2015, suggesting a different seasonal pattern
of PAH exposure to fish in the Mar Menor lagoon.
Acknowledgement
449
This work has been supported by Seneca Foundation (Region of Murcia, Spain) through ‘BIOMARO’ project
(15398/PI/10).
Instituto Español de Oceanografía, Centro Oceanográfico de Murcia; 2Deltares, Marine and Coastal
Systems, PO Box 177, 2600 MH Delft, Netherlands and Institute for Environmental Studies (IVM), VU
University, Amsterdam, Netherlands
1
Extração assistida por micro-ondas de um bioerbicida a partir das folhas
da Canavalia ensiformis no combate da erva daninha Emilia sonchifolia
na cultura de soja
Darlan Ferriera da Silva1; Maria Olímpia de Oliveira Rezende1
O solo é normalmente o destino final dos pesticidas aplicados em diversas
culturas. O destino ambiental desses pesticidas depende das características físicoquímicas especificas de cada composto, além de fatores do próprio ambiente, manejo
do solo e a dose utilizada. Com uma sequência de aplicações, diversos pesticidas
alcançam o solo, devido à aplicação direta e/ou pela lavagem das folhas. Entretanto,
quantidades de pesticidas, particularmente no caso de pesticidas foliares, são
interceptados e absorvidos pelas folhas das ervas daninhas e/ou da própria cultura.
Plantas contaminadas por pesticidas podem retornar ao solo durante o ciclo da
cultura (senescência foliar) ou depois da colheita na forma de resíduos da cultura. O
destino dos resíduos de pesticidas no solo vindos de material vegetal é geralmente
ignorado e desconhecido. O pesticida e seu produto de degradação podem ser
lançados no solo durante a decomposição da planta com uma contribuição lenta
daquelas substâncias para o solo e um adicional risco de contaminação ambiental. A
quantidade de resíduos de herbicidas que irão ser incorporados ao solo depende da
absorção, translocação e degradação dos ingredientes ativos das plantas avaliadas.
Neste sentido, pesticidas obtidos a partir de outras plantas são uma alternativa a
evitar a contaminação do solo.
A extração dos compostos com capacidade herbicida da planta Canavalia
ensiformis foi feita em sistema de microondas, utilizando-se 2 g das folhas secas e
trituradas com 30 mL de água como solvente.
O uso dos extratos aquosos das folhas da Canavalia ensiformis como herbicida,
451
torna-se uma alternativa sustentável de defensivo agrícola, desde sua extração até
Zearalenone as environmental contaminant: occurrence in broa
sua possível biodegradação. Os ensaios realizados em casa de vegetação mostraram
N. M. C. Ribeiro1; L. J. G. Silva1; A. Pena1; C. M. Lino1
que os extratos têm potencial alelopático contra a erva daninha falsa serralha
(Emilia sonchifolia) e que sua absorção e posterior degradação ocorre na planta, não
acarretando danos ao solo.
Zearalenone (ZEA), a metabolite primarily associated with several Fusarium species
(F. culmorum, F. graminearum, F. sporotrichioides, F. cerealis, F. equiseti, F. crookwellense
and F. semitectum) and known as 6-(10-hydroxy-6-oxo-trans-1-undecenyl)-βresorcylic acid-lactone, is a phytoestrogenic compound.ZEA is most commonly
FAPEMA, CNPq
1
Universidade de São Paulo- Instituto de Química de São Carlos (IQSC- USP); darlansilva@iqsc.usp.br;
2
Universidade de São Paulo- Instituto de Química de São Carlos (IQSC- USP); mrezende@iqsc.usp.br
found in maize as well as in its derivatives such as flour.
The European Commission (EC), through EC legislation Nº 1126/2007, set
regulatory limits,as regards Fusarium toxins in maize and maize products, in order
to protect public health and the maximum limit (ML) established for ZEA in bread is
50 µg/kg.
Broa, a traditional maize bread consumed in Portugal has never been studied for
ZEA.
The occurrence of ZEA in maize bread, broa, for human consumption, from the
Portuguese market, was evaluated. Good analytical performance was obtained
through extraction with acetonitrile:water (90:10), clean-up with immunoaffinity
columns, and detection and quantification by liquid chromatography with-tandem
mass spectrometry. ZEA levels were determined in 52 samples to verify the compliance
with the maximum permitted levels by the European legislation. One broa sample
exceeded the ML established byEC. A higher contamination frequency was observed
in samples in which maize and wheat were present. Overall, 13.5% of the samples
were contaminated, at levels oscillating between 9.6 and 50.4 µg/kg. Considering the
percentage of the tolerable daily intake obtained, 0.6%, the risk assessment linked
with the exposure to ZEA was considered very low for the studied population. This
the first study on the intake assessment of ZEA through maize bread consumption
453
in Portugal.
The study highlights the fact that the levels of ZEA in the broa collected from
Portuguese local markets were not totally suitable with regard to the current EC
Free phthalate plasticizer DOPT (dioctylterephthalate) from chemical
recycling of waste pet.
Raul Segura Cruz1; Maria Isabel Velazquez Sandoval2
legislation. It also emphasizes that there was no concern about intake of the ZEA
through its consumption by adult population in the area surveyed.
The objective of this project was to develop the free phthalates plasticizer DOTP
(Dioctylterephthalate) from chemical recycling of PET waste, The project was created
LAQV, REQUIMTE, Group of Bromatology, Pharmacognosy and Analytical Sciences, Faculty of Pharmacy,
University of Coimbra, Polo III, Azinhaga de Stª Comba, 3000-548 Coimbra, Portugal
1
with the objective of replacing the DOP plasticizer still widely used in Mexico.
Plasticizer which is mainly used in PVC applications, conductive coatings,
automobile clothes, glass films, toys, construction industry; pharmaceuticals and
medical devices; and in manufacture of containers and other uses, providing better
performance to be less volatile; more flexible at low temperatures, more stable, and
free phthalate.
In Mexico the use of DOP is still very high, despite the prohibitions for use
internationally to be considered toxic to humans, particularly in liver and testis; and
could be, also has been declared as an endocrine disruptor in Europe. In Mexico, a
study from the National Institute of Public Health and the Cinvestav, concluded that
exposure to phthalates as those contained in the DOP may play a role in the onset of
diabetes in older adults, so the development of materials to replace the use of DOP
is essential.
The first experimental stage consisted of the development high pure terephthalic
acid from waste PET, this stage is based on technology development with
researchers from the Universidad Iberoamericana A.C. in 2014 and wassupported by
theprogramPEI2014 of CONACYT. proj. 213168.
455
The second stage consisted of the development in pilot plant of a novel process
Development of novel coating free VOC´s (volatile organic compounds)
to produce DOTP from terephthalic acid from waste PET, with high specs and
with high anti corrosion properties.
complying with all product specifications.
Maria Isabel Velazquez Sandoval1; Rasul Segura Cruz2; Victor Fermin Gonzalez Saenz3
The main result of our development is the production in pilot plant with more
than 99.5% purity DOTP from PET waste in compliance with specifications of the end
product and substantially reducing the raw material cost of manufacturing to be a
derivative of chemical recycling of PET.
This project allows the development of a new process to produce DOTP from
chemical recycling of PET with a high impact in recovery of PET in higher valueadded materials.
The DOTP project is supported by the program for research, technological
development and innovation 2015 of CONACYT (National Council of Science and
Technology, Mexico).
The main objective of this project was to develop a new free VOC´s (Volatile
Organic Components) water -based coating family from alkyd resin with high
emulsion stability, low solvent, low VOC´s in final application process, Substituting the
use of volatile solvent (typically Mineral spirits, Xylene and Toluene), these solvents
typically are used in 50% by weight in alkyd resins, these solvents are flammable,
petrochemical catalyze the synthesis of Ozone and are neurotoxic. This development
will substantially reduce the VOC´s in the final product; generating a stable coating in
emulsion; with a lower cost formulation and process as well as comparable properties
to the solvent-based coatings, among other benefits, and without affecting film
properties.
Experimental stage consisted of the development at laboratory, pilot plant and
industrial plant scaling formulations and processes to achieve water- dispersible
Grupo Quimico Industrial De Toluca - Grupo Ait Alto Impacto Tecnologico;
Grupo Ait - Rase Proyectos Productivos
1
2
polymer and a stable coating in emulsion. In each stage of development tests such as
viscosity, acid number, total solids, application in coatings, film properties, corrosion
properties, etc.
The main result of our development was the reduction of up to 98% VOC´s in final
coatings with excellence film and anti corrosives properties.
The developed coating has been successfully tested in pilot plant and has clients in
457
use in metal, architectural, structural, automotive two components, traffic paint, etc.
The prototype of free VOC`s coating, is suitable for application in industrial and
commercial applications such as anticorrosive enamel, paint marking and traffic, etc.
Kinetic studies and viability of using peat for removal of Al, Cu and Pb of
an effluent of a plastic recycling company
M. E. K. Carvalho1; C. H. Watanabe1; A. H. Rosa1; D. Goveia2; P. S. Tonello1
In tests in traffic paint application has superior performance against coatings
Introduction: Wastewater can affect the environment when minimum criteria for
based in alkyd and acrylic resins solvent borne and against other waterborne resins.
environmental protection is not followed. Pollution by potentially toxic metals has
caused impacts like bioaccumulation in living organisms and problem of global pollution
technological
through atmospheric deposition. Several studies have shown the potential of peat as
development and innovation2015 of CONACYT(National Councilof Scienceand
metals adsorbent in wastewater. Objective: Analyzing the efficiency of a column filled
Technology,Mexico).
with peat (Santa Amaro das Brotas, Sergipe, Brazil) for removal of Al, Cu, and Pb from an
Thisprojectwas
supported
by
theprogramfor
research,
effluent of a plastic recycling company. Methodology: for kinetics adsorption, solutions
containing all metals, at concentrations between 1 and 20 mgL-1, pH 4.5, received 0.5 g
Tremet - Grupo Ait;
Rase Proyectos Productivos - Grupo Quimico Industrial De Toluca;
3
Grupo Ait - Rase Proyectos Productivos
1
2
of peat. The solutions were filtered through a tangential flow ultrafiltration system with
a membrane of polyethersulfone 1 kDa at predetermined times. Withdrew aliquots of
the filtrate and the concentration of metals was determined by ICP OES. Adsorption
isotherms were constructed using the values ​​of the masses of metals adsorbed by
adsorbent mass as a function of concentration at equilibrium. The collected effluent was
filtered to remove suspended solids and 93 mL of sample was digested to determine the
concentration of metals.
A column of 1 cm diameter and 15 cm in height, with 4g of peat was used to remove
the effluent metals. The original effluent and effluent spiked with concentration of
metals between 0.5 and 10.0 mgL-1 were percolated by the column of peat and aliquots
were collected for analysis of concentration of metals. Results: All elements showed
adsorption between 80 and 100%. Pseudo-second order kinetic was obtained, indicating
chemisorption by peat. Langmuir isotherm was obtained for each metal ion, indicating
formation of homogeneous monolayer on the surface of peat. Conclusion: It can be
459
seen great adsorption of all metals in peat, indicating that a treatment for this type of
Risk Assessment
effluent by a peat column is effective.
Mutagenic activity and indoor air quality in a occupational environment
Environmental Studies Group, São Paulo State University (UNESP), Av. Três de Março, 511, Sorocaba, São
Paulo, Brazil, CEP 18087-180.;
2
Environmental Studies Group, São Paulo State University (UNESP), Rua Geraldo Alckmin, 519, Itapeva, São
Paulo, Brazil, CEP 18409-010.
1
of a images service of a public hospital in Brazil
Fabio Henrique Leite de Oliveira1; Paulo José Balsamo1,3; Elisabeth Palosi Teixeira1,2; Silvia Pierre
Irazusta1,2;
In general, the health effects related to the indoor pollutants can be classified in
two categories: those that appear immediately after exposure, and those that appear
only after many years. In hospitals,there is a problem with the use of radiation that is
quite common in diagnostic imaging industry, since it is not known with precision,
the biological effects of exposure to residual levels of radiation for prolonged
periods, both for the development of a lesion (somatic), as well as to cause
mutation (genetic level). This work aimed to access the radiation safety through the
monitoring of the effects of possible residual radiation in the workplace of exposed
professionals in a Service of Images of a Public Hospital Complex in the Sorocaba
City- São Paulo State- Brazil, by means of a bioindicator. Thus, we intend to further
clarify to employees, technicians and radiologists about the risks to which they are
exposed in the exercise of their activity. The micronucleus assay with Tradescantia
spp. (Trad-MCN) is considered one of the most sensitive and efficient for detection of
mutagenic agents in the air was the bioassay chosen for this evaluation. This study
aimed at also, the application of a questionnaire to verify the level of knowledge
of employees on biosafety and identification of airborne fungi in the environment.
The results showed that residual radiation levels at the sites studied were able to
induce mutagenicity, i.e., the data indicate that there are physical hazards related
to the residual radiation in the evaluated areas, except for the densitometry. Most
employees who responded to the questionnaire said they had knowledge of the
461
rules set out in resolution and claim properly perform their activities in relation to
Metais no Corpo Aquático do Riacho Mussuré e o córrego Mumbaba e o reflexo
the safety standards. The macro and microscopic analysis of the samples showed
de sua degradação na saúde dos ribeirinhos - João Pessoa - Paraíba – Brasil
fungal spores of the genera Aspergillus, Penicillium, Cladosporium, Fusarium and
M. L. P. da Silva1; J. D. Neto2
Nigrospora. Despite the knowledge of radiation protection standards, the risk of
residual levels of radiation are unknown and prevention measures are multifactorial.
O trabalho abordar a problemática dos resíduos tóxicos das indústrias que são
lançados no Riacho Mussuré e no Córrego em Mumbaba, oriundos do Distrito
Industrial João Pessoa – PB/Brasil, que provocam alterações diversas na qualidade da
1.Faculty of Technology of Sorocaba - SP - Brazil; 2.Postgraduate Program in Management and Technology
in Production Systems - State Center of Technical Education in the State of Sao Paulo –CEETEPS- Brazil;
3Federal University of São Carlos-SP-UFSCar- Brazil.
água, na cadeia alimentar da biótica aquática, na saúde da comunidade ribeirinha.
Objetivo: Investigar os impactos do lançamento de efluentes industriais e domésticos
na qualidade das águas do córrego e do riacho Mussuré e as consequências na saúde
dos ribeirinhos. O riacho Mussuré, inserido na bacia do rio Gramame, João Pessoa –
PB/Brasil. E o córrego localizado na comunidade Mumbaba no Município de João
Pessoa – PB/Brasil. Ambos permeiam a comunidade Mumbaba. Foram coletadas
amostras de água em cinco pontos diferentes, no Córrego e no riacho, de janeiro a
dezembro de 2014. As variáveis temperatura, oxigênio dissolvido, pH, DBO, turbidez,
condutividade elétrica e coliformes termotolerantes, analisadas no laboratório da
Superintendência de Administração do Meio Ambiente – João Pessoa – PB/Brasil.
Os metais cromo, chumbo, níquel, cádmio, cobre e mercúrios foram analisados no
Instituto de Tecnologia de Pernambuco–Recife/Brasil, utilizando-se espectrometria
de emissão com plasma indutivamente acoplado (ICP-OES). Foi aplicado um
questionário em 130 domicílios, com amostra de 680 indivíduos. As águas do riacho
Mussuré apresentam teores de metais, cromo 0,06 Mg/L, cobre 0,05 Mg/L, níquel
0,03 Mg/L acima do valor permitido da Resolução CONOMA 357/2005. A Demanda
Bioquímica de Oxigênio das Águas do córrego apresentam valores elevados atingindo
160,0 mg/L. Em todos os pontos de coleta, oxigênio dissolvido, apresentam baixas
concentrações, variou 0,3 até 2,8 mg/L. Coliformes termotolerantes, apresentam
463
valores altos que chegam a 1,93 x 105col/mL. Nos resultados epidemiológicos
Water treatment by nanofiltration and reverse osmosis: chemical and
as doenças mais frequentes na população são as doenças respiratórias e doenças
ecotoxicological efficiency
hídricas. Conclui-se que o riacho Mussuré e o córrego de Mumbaba apresentam alto
Patrícia Palma1; Sofia Fiallho2; Cristelle Santos3; Gabriela Palma4; Carla Cavaco4; Rui Gomes4; Luisa Neves5
grau de degradação e suas águas representam um risco para a saúde da população.
The increment of world´s population with the consequent demand for water
supply, plus the sustainable development and the conservation of water resource will
Universidade Federal de Campina Grande UFCG-PB
1
require the reuse, recycling and the improvement quality of wastewater discharges.
Within this context, this work investigated the potential of using membrane
technology, in particular nanofiltration and reverse osmosis membranes to ensure
the quality of treated water. The aim of the present study was to characterize and
evaluate the efficiency of nanofiltration (NF90, NF270) and reverse osmosis (SW30)
membranes, in improving the quality of three types of waters: irrigation, supply and
wastewater.The efficiency of using this technology was assessed by: (i) the study of
water permeability and rejection of chemical parameters such as total nitrogen, total
phosphorus, carbon organic demand (COD), biochemical oxygen demand (BOD5),
feacal coliforms, nitrates, chlorides, iron and manganese; and (ii) the evaluation
of ecotoxicological effects through the use of representative species of different
taxonomic and functional groups (Vibrio fischeri, Thamnocephalus platyurus,
Daphnia magna, Pseudokirchneriella subcapitata). The results indicated that the
three membranes have high rejection percentages for the most parameters studied
(>90%). All parameters concentrations in the permeate stream were found to be
below the legal limits, according to the Decree-Law Nº. 306/2007 of 27 August and
Nº. 236/1998 of 1 August. With respect to the ecotoxicological effects, in general, the
use of membranes removed the toxicity detected in the initial samples (classified as
Class 2 of UT). The SW30 membrane showed a greater toxicological efficiency removal
comparing with the nanofiltration membranes. Further, among the nanofiltration
465
membranes used, the NF90 promoted a greater decrease in the toxicity of the
Pesticide mancozeb®: derivation of risk limits for freshwater ecosystems
samples. Thus, treatment with the membranes NF90 and SW30 allowed obtaining
M. Samussone1; M.N.Vieira1; A.P.Carvalho1; R. Pereira1
waters with high quality and low ecotoxicological potential, without risks for human
Agriculture is a sector that ensures the sustainability of the world´s largest economies
populations and endangering the environmental balance.
and that guarantees poverty reduction. However, the intensive use of pesticides as a
DTCA - Instituto Politécnico de Beja, Rua Pedro Soares S/N, Apartado 6155, 7800-295 Beja, Portugal/
Centro de Investigação Marinha e Ambiental, FCT, Edifício 7, Piso 1, Universidade do Algarve;
2
DTCA - Instituto Politécnico de Beja, Rua Pedro Soares S/N, Apartado 6155, 7800-295 Beja, Portugal;
3
Centro de Biotecnologia Agrícola e Agro-Alimentar do Alentejo; 4Empresa Municipal de Água e
Saneamento de Beja; 53Centro de Biotecnologia Agrícola e Agro-Alimentar do Alentejo/5REQUIMTE/
CQFB, FCT, Universidade Nova de Lisboa
1
means of crop protection has generated numerous problems at the ecosystem level
due to drainage of agro-chemical residues into aquatic ecosystems, with impacts on
non-target species, including humans. In this context, the present study aimed to assess
the ecotoxicological effects to aquatic organisms of the fungicide Mancozan® (active
ingredient Mancozeb®), and determine the limits of risk for these ecosystems. This
pesticide belongs to the group of ethylene-bis (dithiocarbamate), and is widely used
both in Europe and Mozambique. A battery of ecotoxicological assays with organisms of
different trophic levels including the bacterium Vibrio fisheri, the green alga Raphidocellis
subcapitata, the aquatic plant Lemna minor, the cladoceran Daphnia magna, and the
freshwater fish Danio rerio and Carassius auratus, were performed. The tests followed
the standard OECD protocols, except for the evaluation of the activity of the enzyme
acetylcholinesterase in Carassius auratus. The animal species were the most sensitive.
The pesticide showed high toxicity at concentrations much lower than those applied in
the environment. Based on the results obtained a value of PNEC (Predicted No Effect
Concentration) of 0.024 mg L-1 was estimated for Mancozeb®. This work also contributed
for transfer of knowledge in risks assessment methodologies, already established in
Europe, in order to promote its implementation in African countries.
Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR/CIMAR), Universidade do Porto,
Rua dos Bragas 289, 4050-123 Porto, Portugal; Departamento de Biologia da Faculdade de Ciências da
Universidade do Porto, Rua do Campo Alegre s/n 4169-007 P
1
467
Keyword:
Ionizing radiation, biosecurity, Tradescantia pallida, mutagenicity, airborne fungi
Histopathological analysis of livers from barbel (Luciobarbus bocagei), nase
(Pseudochondrostoma duriense) and trout (Salmo trutta) from Douro basin
J. Carrola1; D. Santos2; A. Pires3; S.M. Monteiro1; A.M. Coimbra1; S. Oliveira1; V. Pereira1; C. Santos1; J. Jesus1;
M. Lopes1; H. Neves2; S. Hughes1; R. Cortes1
The effective assessment of the ecological status of the streams and rivers is
increasingly important or even mandatory. Fish histopathological biomarkers,
namely hepatic histopathology, represent a valuable tool that can be used to assess
pollution effects.
This study evaluates the prevalence and gradient of hepatic alterations/lesions,
using an eight-grade scale: 0) normal to 7) severe changes. During the summer of
2014, barbel (Luciobarbus bocagei), nase (Pseudochondrostoma duriense) and trout
(Salmo trutta) were captured at 4 sites covering a quality gradient in the Douro basin:
Corgo River, in Alvações do Corgo (L1); Paiva River, in Castro d’ Aire (L2); Bestança
River, in Covelas (L3) and Paiva River, in Ameixiosa (L4).
Histopathological analysis of fish livers revealed very low level hepatic changes/
lesions in fish from the study sites. The main lesions observed presented low severity
for fish health and were lymphocytic foci (LF), macrophage aggregates (MA),
granulomas (GR), inflammatory foci (IF), glycogen accumulation (GA), necrosis (NE),
and pyknosis (PK). Trouts were caught only in L3, and presented the lowest gradient
values. Nase were caught at all sites and showed the highest gradient values. The
average gradient value of all lesions in nase was 0.071 in L1 (site classed as Moderate
Ecological Status) and statistical significant differences (P=0.007) were detected with
sites L2 and L3 (both with 0.018, classed as Good and Excellent, respectively).
Liver lesion gradient detected in the nase showed that this is the best fish species
for histopathological detection of ecological status. However, barbel were not found
in L3 and were present in low numbers in L2 and L4, whereas trout was present only
469
in L3. Despite that, the data point to a low level of pollution at all sites and indicated
Adjusting molecular methods to the detection of toxigenic phytoplank-
that the gradient average of all liver lesions (in a single species) is the best indicator
ton genotypes in environmental and preserved samples
of Ecological Status.
C. Diniz1; P. Palma1,2; M.P. Reis1
Acknowledgements:
“This work is supported by the Programa Operacional Regional do Norte (ON.2 – O Novo Norte), through
the Fundo Europeu de Desenvolvimento Regional (FEDER) and Fundação para a Ciência e a Tecnologia
(FCT/MEC), under the project SUSTAINSYS: NORTE-07-0124-FEDER-0000044.”
Centre for the Research and Technology for Agro-Environmental and Biological Sciences (CITAB),
University of Trás-os-Montes and Alto Douro (UTAD), Quinta de Prados 5000-801 Vila Real, Portugal; 2Life
Sciences and Environment School, University of Trás-os-Montes and Alto Douro (UTAD), Quinta de Prados
5000-801 Vila Real, Portugal; 3Mathematic Department, University of Trás-os-Montes and Alto Douro
(UTAD), Quinta de Prados 5000-801 Vila Real, Portugal
1
Cyanobacteria are aquatic photosynthetic bacteria that may form dense blooms,
known as Cyanobacteria Harmful Algal Blooms (CyanoHABs). These decrease water
quality, may accumulate surface scums and originate toxicity events, since many
species can produce harmful, biologically active substances. Cyanotoxins, like
cylindrospermopsins (CYN) or saxitoxins (STX), may be quantified through ELISA
assays, HPLC or LC-Ms/Ms, methods that lack cost effectiveness for regular use in toxicity
screening. Such toxins have a wide range of toxicity mechanisms, with hepatotoxic,
neurotoxic, cytotoxic and even genotoxic effects, and may thus be detected through
the use of ecotoxicological assays. In fact, assays testing Thamnocephalus platyurus
mortality or the inhibition of mobility and/or reproduction of Daphnia magna are
known to successfully detect CYN and SXT toxicity. The presence of toxigenic species
may also be assessed through the screening for toxin production genes among
phytoplankton DNA. CYN biosynthesis is encoded by the cyr gene cluster and STX
depends on the sxt gene cluster. Toxigenic genera, producing either CYN or STX
have been detected in many environments. Thus, the main goal of this work was
to evaluate the association of ecotoxicological tools with the detection of toxigenic
cyr or stx genotypes, in order to assess environmental risks linked to CyanoHABs.
We used DNA extracted from fresh and preserved environmental samples, as well as
from cultured isolates in PCR amplification, testing different sets of specific primers
for cyr or stx. The presence of specific amplicons for these genes was compared with
acquired ecotoxicological data. While transcriptomic detection of the expression of
cyr or sxt operons is still not cost effective, and gene detection doesn’t guarantee
471
their actual expression, the association of ecotoxicological tools to the detection of
Cyanobacteria prevalence and cyanotoxin occurrence in an oligotrophic
toxigenic genotypes seems the most effective procedure to help predict risks linked
freshwater reservoir (Beliche – Southern Portugal)
to toxicity of CyanoHABs events.
S. Caetano1,2,3; C. Costa2,3; C. Gago4; M. Rodrigues2; C. Mateus5; M.P. Reis3
Center for Marine and Environmental Research (CIMA), Universidade do Algarve, edifício 7, Campus de
Gambelas, 8005-139 Faro, Portugal. E-mail: mpreis@ualg.pt
2
Department of Technologies and Applied Sciences, Polytechnic Institute of Beja – School of Agriculture,
7800-295, Beja, Portugal
1
This study aimed to assess the prevalence of cyanobacteria and their toxins in an
oligotrophic reservoir that supplies the Eastern Algarve population with urban and
irrigation water. Despite having lower maximal growth rates than the majority of other
phytoplankton species, cyanobacteria are able, in certain conditions, to overtake all
other species, resulting in cyanobacteria blooms (CyanoHABs), even in oligotrophic
systems. Although cyanotoxins presence is more often observed in nutrient enriched
environments, such metabolites are also produced in oligotrophic systems, though
usually in lower concentrations. Samples were collected on a monthly basis during
9 years, from 2003 to 2011. Analysis of phytoplankton and supporting physical and
chemical variables were performed according to European standard protocols.
Nutrient concentrations confirmed oligotrophy and chlorophyll-a geometric mean
of 1.1 µg/L was also bellow the upper limit of oligotrophy (2.5 µg/L). High molar
DIN:SRP ratios, well above the Redfield-Richards ratio of 16:1, also contradict some
literature on favouring conditions for cyanobacteria development. Yet, cyanobacteria
dominated in abundance during most of the time and sometimes also in terms of
biovolume. In fact 62.1% of the samples had abundances of cyanobacteria above the
WHO alert level 1 (2000 cells/ml) reaching maximum of 1.3x105 cells/ml, also above
the alert level 2 (105 cells/ml). Some of these CyanoHABs were mainly composed
by potentially toxic genera, such as Anabaena, Microcystis or Oscillatoria, and in a
few cases cyanotoxins were also detected, only surpassing the WHO safety guide
level of 1 µg/L in bottom samples taken during the summer 2006, after a prolonged
drought. As observed for other reservoirs, in Beliche cyanobacteria slow growth
473
rates seem to be compensated by high prevalence of populations, selected by warm
Gill histopathological evaluation in fish species captured in the Douro
summer waters, which inhibit growth of other phytoplankton groups. The fact that
river basin
the predominant genera were potentially toxigenic constitutes an environmental
Marta Fraga1; Ana Maria Coimbra2; João Carrola2; Rui Cortes2; Samantha Hughes2; Simone Varandas2; Vitor Pereira2; Cátia Santos2; Marisa Lopes2; Dércia Santos1; Sandra Mariza Monteiro2
and public health concern.
School of Health, University of Algarve, Avenida Dr. Adelino da Palma Carlos, 8000-510 Faro, Portugal
AquaExam, Lda., Centro Empresarial de Gambelas, Pavilhão B1, Universidade do Algarve, 8005-139 Faro,
Portugal
3
Center for Marine and Environmental Research (CIMA), Universidade do Algarve, edifício 7, Campus de
Gambelas, 8005-139 Faro, Portugal. E-mail: mpreis@ualg.pt
4
Agência Portuguesa do Ambiente (APA- ARH Algarve, I.P.), Rua do Alportel, nº 10, 2º, 8000-293 Faro,
Portugal
5
Chemistry Research Center of Algarve (CIQA), Universidade do Algarve, edifício 2, Campus de Gambelas,
8005-139 Faro, Portugal
1
2
The hydrographic Douro basin is a strategic area for water supply, agroforestry
landscape and hydroelectric energy production. Therefore, the ecological status
assessment of this region streams is critical for an environmental management.
Fish gill histopathology is known to reflect gradients of water contamination, being
a biomarker of ecosystems disturbance, what potentiates its use in monitoring
programs. In the present study, in order to identify possible associations with
environmental descriptors and ecological status classifications, gill histopathological
changes were quantified in fish captured in four locations of the Douro basin:
Corgo River, in Alvações do Corgo (L1); Paiva River, in Castro d’Aire (L2); Bestança
River, in Covelas (L3) and Paiva River, in Ameixiosa (L4). In these catchments, two
native species were captured: nase (Pseudochondrostoma duriense) and barbel
(Luciobarbus bocagei).
Data analyses revealed that, in all locations, the captured fish showed some degree
of gill histopathological changes. The main changes observed were aneurisms (AN),
epithelial lifting (LFT), vasodilatation (VD), edema (ED), lamellar fusion (LF), lamellar
epithelium proliferation (LEP), filament epithelium proliferation (FEP) and necrosis
(N). The one with higher prevalence was LEP, while the most unusual was necrosis.
In barbel, no statistical differences were observed among sites in the lesions severity
degree. In nase, a significantly higher degree of FEP was observed in L1 (classified
with a Moderate Ecological Status), in comparison with L3 and L4 (both classified
with Excellent and considered as reference sites). Likewise, a significant higher
degree of LEP was observed in L1, comparatively to L3. The severity degree of the
475
remaining histopathological changes, showed no statistical differences among sites.
Endocrine Disruption
Despite this, in both fish species, higher levels of severity were observed in disturbed
sites, L1 and L2 (classified with Good Ecological Status). In general, data suggests that
Chronic toxicity of the antiepileptic drug carbamazepine on crustacean
the severity gradation scale of the gill histopathological changes, increases with the
Daphnia magna.
disturbance level, confirming the ecological status classification and proving to be a
A. Oropesa1*; A.M., Floro2; J., Guerreiro2; P., Palma2,3
useful tool to integrate biomonitoring programs.
Acknowledgements:
“This work is supported by the Programa Operacional Regional do Norte (ON.2 – O Novo Norte), through
the Fundo Europeu de Desenvolvimento Regional (FEDER) and Fundação para a Ciência e a Tecnologia
(FCT/MEC), under the project SUSTAINSYS: NORTE-07-0124-FEDER-0000044.”
Life Sciences and Environment School, University of Trás-os-Montes and Alto Douro (UTAD), Quinta de
Prados 5000-801 Vila Real, Portugal; 2Centre for the Research and Technology for Agro-Environmental and
Biological Sciences (CITAB), University of Trás-os-Montes and Alto Douro (UTAD), Quinta de Prados
5000-801 Vila Real, Portugal
1
Trace quantities of pharmaceuticals are continuously being discharged into the
environment through domestic and industrial wastewater effluents resulting in the
chronic exposure of aquatic organisms. Despite the ubiquity and recalcitrance of
carbamazepine (CBZ), an antiepileptic drug, its chronic effects in crustacean Daphnia
magna are unknown.
The aim of this study was to evaluate the chronic toxicity induced by CBZ (10,
100 and 200 µgL-1) in the life cycle, embryo development and sex differentiation
of D. magna. The endpoints used were fecundity (number of offspring per female),
percentage of male production, developmental abnormalities and growth of females,
through the development of the chronic reproduction bioassay and the embryo
toxicity test. Further, the possible anti-juvenoid activity of the drug was assess in the
presence of fenoxycarb (FEN, 1 µgL-1 positive control), an insect growth regulator,
that mimics the action of methylfarnesoate and promoted the production of 95% of
male offspring at the given concentration.
CBZ and CBZ+FEN caused a concentration-dependent decrease in the number of
offspring produced per female comparing to the control groups (ASTM+DMSO). In
addition, the offspring production, the moulting frequency and the size of females
showed a more accentuated decrease in the mixtures (CBZ+FEN). Production of
males could be observed in all CBZ and CBZ+FEN concentrations. No toxic effects on
477
the embryonic development were observed after the exposure of embryos to CBZ
Monitoring and Modelling
and CBZ+FEN concentrations. Therefore, the results suggest that CBZ interferes with
the life cycle and sex determination of the crustacean D. magna.
Modelización espacial de la contaminación por nitrato en los acuíferos
de la Cuenca Alta del río Ebro (España) mediante SIG
Authors wish to thanks the financial support given to A.L. Oropesa by Ministerio
Mercedes Arauzo Sánchez1
de Educación, Cultura y Deporte en el marco del Programa Estatal de Promoción del
Talento y su Empleabilidad en I+D+i, Subprograma Estatal de Movilidad, del Plan
Estatal de Investigación Científica y Técnica y de Innovación 2013-2016 through a
El nitrato constituye la principal fuente de contaminación difusa de los recursos
post-doctoral research grant (CAS14-00224), also to “Consejería de Empleo, Empresa
hídricos del planeta. La contaminación surge cuando una masa de agua (superficial
e Innovación. Gobierno de Extremadura” and the Feder funds for the economic
o subterránea) recibe aguas de infiltración o escorrentía cargadas de compuestos
support.
nitrogenados, generalmente de origen no natural.
El estudio se desarrolló en la cuenca alta del Ebro (25.664 km). El área de campo
se encuentra dividida en 22 cuencas hidrográficas, que drenan hacia un total de 46
Unidad de Toxicología. Departamento de Sanidad Animal. Facultad de Ciencias. Universidad de
Extremadura, Badajoz 06071, España.
2
Departamento de Tecnologias e Ciências Aplicadas. Escola Superior Agrária de Beja, Beja 7801-295,
Portugal.
3
CIMA-Centro de Investigação Marinha e Ambiental, CIMA, FCT, Universidade do Algarve, Faro 8005-139,
Portugal.
*e-mail (correspondig author): aoropesa@unex.es
1
masas de agua subterránea (18.704 km2).
La modelización del contenido en nitrato en las aguas subterráneas mediante
funciones de Análisis Espacial de ArcGIS 10, reveló la existencia de 44 zonas afectadas
por la contaminación ([NO3-] ≥ 50 mgL-1) en 17 masas de agua (9 acuíferos aluviales, 8
carbonatados y detrítico-carbonatados). Otras 5 masas de agua mostraron riesgo de
contaminación ([NO3-] ≥ 25-49 mg L-1; 1 acuífero aluvial y 4 detrítico-carbonatados).
Las 24 masas restantes (52% del total) presentaron niveles bajos o moderados de
nitrato.
El 90% de los acuíferos aluviales apareció afectado por la contaminación y el
10% restante en riesgo. Los aluviales con mayor grado de afectación (59-100% de
su superficie) pertenecían a cuencas de primer y segundo orden. El 60% presentó
correlaciones negativas entre la concentración de nitrato y la cota piezométrica,
revelando la existencia de “zonas sumidero”, donde el nitrato se acumula en cotas
479
bajas como consecuencia del flujo advectivo descendente. En los aluviales del cauce
Modelización espacial de los riesgos asociados a usos del territorio que
principal del Ebro (con cuencas de drenaje de cuarto y quinto orden) no se observó
afectan a la contaminación por nitrato en la cuenca alta del ebro (España)
significación, debido a su mayor complejidad desde el punto de vista hidrológico.
Mercedes Arauzo Sánchez1
El 22% de los acuíferos carbonatados y detrítico-carbonatados se mostró
contaminado por nitrato y el 11% alcanzó niveles de riesgo. De estos, el 50% mostró
El nitrato es el principal contaminante de origen difuso que afecta a los recursos
correlaciones negativas entre la concentración de nitrato y la cota piezométrica,
hídricos de la cuenca del río Ebro (España). Para poder evaluar la vulnerabilidad a la
revelando nuevamente la existencia de “zonas sumidero”.
contaminación por nitrato en sus diferentes masas de agua, es esencial realizar una
valoración previa de los riesgos asociados a los usos del territorio, susceptibles de
dispersar compuestos nitrogenados en exceso sobre la superficie topográfica.
Instituto de Ciencias Agrarias, Consejo Superior de Investigaciones Científicas
1
El objetivo del estudio se centró en desarrollar modelizaciones espaciales de
los riesgos asociados a los usos del territorio, fundamentalmente agricultura de
regadío y secano, ganadería intensiva y extensiva, y presencia de áreas urbanas,
mediante el uso del SIG ArcGIS 10.0. Los datos de partida fueron el Mapa de Cultivos
y Aprovechamientos de España en formato digital, escala 1:50.000, y los censos
ganaderos en producción intensiva y extensiva de 2008 por municipio, en formato
Excel (Ministerio de Agricultura, Alimentación y Medio Ambiente 2009).
El estudio se desarrolló en la cuenca alta del río Ebro (25.664 km). El área de campo
se encuentra dividida en 22 cuencas hidrográficas, que drenan hacia un total de 46
masas de agua subterránea (el 48% presenta zonas contaminadas por nitrato, o en
riesgo).
El mapa del Riesgo Asociado a los Usos del Territorio reveló una clara relación entre
las zonas de agricultura en regadío y las áreas aluviales (con riesgo alto y extremo;
las más contaminadas), si bien también se apreciaron riesgos atribuibles al regadío
y al secano en extensas superficies sobre acuíferos carbonatados. Por otra parte, la
presencia de bosque y área natural (con usos no contaminantes) podría considerarse
un factor de protección de los recursos hídricos ante los procesos de lixiviación de
481
contaminantes en las áreas de montaña y cabeceras de cuenca.
De los mapas de Riesgo Asociado a la Ganadería Intensiva y a la Ganadería
Extensiva se desprende que, en principio, no es posible atribuir riesgos significativos
Monitorização e integração de fatores ambientais na modelação de
castas portuguesas”
Ricardo Costa1; Helder Fraga1; Aureliano C. Malheiro1; José Moutinho-Pereira1; João A. Santos1
asociados a estas actividades, por lo que cabe interpretar que la gestión de los
residuos ganaderos en el área de estudio se realiza, en términos generales, de modo
sostenible.
O sector vitivinícola tem uma grande importância na economia Nacional
Portuguesa, contribuindo com quase 50% de exportações no total de vinho
Instituto de Ciencias Agrarias, Consejo Superior de Investigaciones Científicas
1
produzido. Portugal encontra-se dividido em 14 regiões vitivinícolas com 25
denominações de origem, sendo as mais importantes as regiões do Douro/Porto,
Vinho Verde, Alentejo e Lisboa. Um dos fatores que exercem influência na produção
vitivinícola é o clima, visto exercer um grande impacto no desenvolvimento
fenológico, bem como nas características do produto final, o vinho. Assim sendo
torna-se evidente que ferramentas que possibilitem previsões dos parâmetros de
produtividade vitivinícola, são de grande importância para o sector, permitindo o
planeamento, monitorização e adaptação de técnicas culturais a curto e longo prazo.
Neste estudo foi utilizado o modelo dinâmico STICS (Simulateur mulTIdisciplinaire
pour les Cultures), adaptado para as castas e condições de desenvolvimento
existentes em Portugal. Para tal, foram efetuadas medições dos parâmetros
específicos das castas Portuguesas, e também dos parâmetros específicos do clima,
solo e praticas culturais existentes. Para este caso de estudo foram utilizadas as
variedades Aragonez, Touriga-Franca e Touriga Nacional, que representam as três
mais importantes variedades em Portugal. Os resultados deste estudo mostram
que é possível a adaptação do modelo STICS à vinha, para a simulação dos estados
fenológicos, produção e stress hídrico. No entanto, o modelo não demonstrou ser
eficaz na simulação do conteúdo em álcool potencial da uva. Os resultados obtidos
através da utilização deste modelo dinâmico de culturas permitirão aumentar a
483
sustentabilidade do sector vitivinícola em Portugal, tornando-o mais competitivo.
Assessment of freshwater bivalves’ communities in Mira lagoons
T. Assunção; S. Varandas1; A. Teixeira2
Centre for Research and Technology of Agro-Environment and Biological Sciences (CITAB), Universidade
de Trás-os-Montes e Alto Douro, 5000-801 Vila Real, Portugal
1
One of the most threatened faunistic groups worldwide, and in overall decline,
are the Unionidae (Mollusca), the largest of six families of freshwater mussels. For
many species (regardless of their conservation status), the most basic data about its
development cycle, size and population structure are worldwide scarce and, in many
cases, non-existent.
Since, until now, the studies carried out with freshwater bivalves in Portugal
concern almost exclusively to the river systems (lakes have been forgotten by their
little representation / expression at the national level), we tried to fill this gap with this
study. Thus, the survey of freshwater bivalve community and the evaluation of the
disturbance level is essential to strengthen the Priority Site status Conservation that
the lagoons already have. This work aims to provide a consolidated set of knowledge
to the community for the natural values of the lagoons as well as to alert for the
importance of controlling invasive species in the preservation and development
of biodiversity through the conservation of freshwater bivalves, their habitats and
hosts.
The results show the presence of three species of freshwater bivalves in both
lagoons an exotic species (Corbicula fluminea) and two native species (Anodonta
cygnea and Unio delphinus) listed on is the IUCN Red List of non-marine molluscs
as Near Threatened (Cuttelod et al., 2011). It was also possible to verify that both
lagoons have a good rating for the Ecological Status, complying with the parameters
required by the Water Framework Directive for reservoirs.
485
Keywords
Freshwater bivalves, development cycle, conservation, hosts.
CITAB-UTAD: Centro de Investigação e de Tecnologias Agro-Ambientais (CITAB), Universidade de Trás-osMontes e Alto Douro (UTAD), Apartado 1013, 5000-801 Vila Real, Portugal;
2
CIMO-ESAIPB–Centro de Montanha, Escola Superior Agrária, Instituto Politécnico de Bragança, Campus
de Santa Apolónia, Apartado 1172, 5301-854 Bragança, Portugal
1
Patrocinadores / Sponsors