LEVEL 1

MONITORING OF CYANOBACTERIA BLOOMS IN
FRANCE AND SOME OTHER EUROPEAN
COUNTRIES
(Germany, Portugal and United-Kingdom)
Hélène DUCOBU
CRITT Bio-Industries Midi-Pyrénées-CRT
INSA
135, Avenue de Rangueil
31077 Toulouse Cedex 4
FRANCE
Tel: +33 (0)5 61 25 35 57
Fax: +33 (0)5 61 55 94 00
E-mail: helene.ducobu@wanadoo.fr
•
France is divided into 22 régions and 96 departments
And 6 hydrographic basins
Water monitoring concerns several structures which
act at different administrative levels
LEVEL 1: NATIONAL
Ministry of the Environment (Ministry
of Ecology and sustainable Development)
Mission: Preservation and protection
of the water resource
Ministry of the Health (Ministry of
Health, Family and handicapped People)
Mission: Sanitary protection of the
water users
LEVEL 2: Hydrographic Basin
Water Agencies* (6)
LEVEL 3: REGIONAL
DIREN* (regional direction of the
environment)
* Field work
DRASS (regional direction of the
sanitary and social affairs)
LEVEL 4: DEPARTEMENTAL
DDASS* (departmental direction of the
social and sanitary affairs)
MONITORING OF CYANOTOXINS.
From awareness to reglementation
1999: Program EFFLOCYA initiated by the Ministry of the Environment and
the Ministry of the Health. Collaboration with scientists (among which the
Pasteur Institute). Step I: An inquiry sent to the Water Agencies, DDASS,
Universities… to evaluate the occurrence of algae blooms in France. Simple
questions such as: « Did you observe a change in lake’s colour? What was the
colour (green, blue-green or red)? Did you notice dead animals? During which
month of the year did the bloom occur? Did you make an observation under
the microscope? …
RESULTS:
Low rate of reply (only 30 %) but it was a first step in the awareness
process.
Interesting results: The blooms occurred during Summer mainly in water
reservoirs used for bathing or fishing. Most of the blooms were green in
colour. Microcystis and Anabaena appeared to be the dominant
cyanobacteria. Cases of mortality have been deplored among fishes and
birds. Almost all the departments were concerned by the problem.
Step II: Measuring
Work of a small group of laboratories. Presence of toxins in several lakes. Lack
of data in some regions of France (South). Lack of collaboration and
communication between the laboratories. Almost everything occurred in Paris!
ÎAll the scientists working on cyanobacteria decided to meet each others to
exchange their experiences and to federate.
2001: Creation of the GIS (Group of Scientist Interest) CYANOBACTERIA.
At the same time, the EFFLOCYA program did stop…
Partners of the GIS:
Research Institutes: INRA, CNRS; Pasteur Institute of Paris; Museum of
History of Natural Sciences of Paris; Commissariat of Atomic Energy (CEA)
French Agency for Sanitary and Alimentary Security (AFSSA); Universities;
CRITT Bio-Industries Midi-Pyrénées...
Main objectives:
ÎResearch (determinism of cyanobacterial blooms; molecular tools);
ÎMonitoring (standardization of the methods, development of new tools);
ÎRisk assessment;
ÎRemedial measures;
ÎEvaluation of the socio-economic impacts;
ÎIndustrial valorisation of cyanobacteria;
ÎCommunication (interface between managers and scientists).
REGLEMENTATION
Brazil: First country to propose and implement a
reglementation at a national level (Portaria n°1-469;
2000).
World Health Organization: Recommandations for
drinking water (1999) and bathing waters (2000).
Guide Value: 1 µg/l MCYST LR
European Community:
Many directives concerning water
quality but none about the control of cyanobacteria. But a
standardization of the protocoles is starting at a European
level.
FRANCE: Order in Council n° 2001-1220 about water for
human consumption (in application at the end of 2003)
includes new parameters such as MCYST LR.
Recommandation DGS/SD7a-N°2003-270 (June 2003, in
application immediatly) about the control of cyanobacteria and
toxins in bathing waters.
WHO RECOMMANDATIONS
Alert levels
Drinking water
Bathing water
Cels cyanos/ml
Vigilance
Level
Actions
Alert 1
Level
µg Chla Cyanos/l
200
2000
20 000
Alert 2
Level
TOXINES
100 000
0.1
1
10
0.2-1 µg/l
Guidance
Level 1
2-10 µg/l
Actions
50
Niveau
guide 2
20-50
µg/l
MONITORING OF TOXIC CYANOBACTERIA IN
COLLABORATION WITH 5 WATER AGENCIES
(2000-2003)
Artois-Picardie
Rhin-Meuse
Loire-Bretagne
Adour-Garonne
Rhône-Méditerrannée-Corse
AIMS OF THIS STUDY ?
It was the very first time that the Water Agencies participated to a
monitoring of toxic cyanobacteria
• Complete the EFFLOCYA data. Especially data missing in the South.
•Identify and list the water bodies which represent a risk and that
should be monitored regularly.
•To test and optimize the protocoles.
•To think about the possibilities of a long-term monitoring (sampling
frequency, with which actors, who is going to finance?…).
•To make the water managers and water users aware of this
environment and health problem.
•To think about the way to communicate the results (to inform
without causing panic).
HOW DID WE START TO WORK?
•January 2000: At this time, the 6 Water Agencies had received the
EFFLOCYA inquiry en did have some data about the occurrence of
cyanobacteria blooms in the water bodies belonging to each basin.
•March 2000: I organized for the 6 Water Agencies a seminary about toxic
cyanobacteria and made them a practical and financial proposal for a
screening of toxic blooms in the 6 hydrographic basins during the Summer
2000. The Water Agencies were asked to finance the study totally.
•May 2000: 3 Water Agencies joined immediately. I had to work hard to
convince the 2 other ones. And the last one refused for financial reasons.
Not possible to get a financing « Inter-Agence » because of administrative
delay. So, each Water Agency did finance a study on its own territory and
did chose its own sampling frequency (lack of homogeneity).
•June 2000: Elaboration and distribution of the protocoles. I elaborated
them together with Ingrid Chorus, in Germany who was used to work with
Water Agencies. Some Water Agencies did ask for a training about sampling
(we organized a one day training). We also sent to each Agency field and
sample sheets and specific containers for samples.
•Summer 2000: Sampling by Water Agencies and by Bi-Eau for the Water
Agency Loire-Bretagne. Sending of frozen material (Chla and toxins) and
non-frozen material (algal determination) to our laboratory.
How the Water Agencies did chose the water bodies to sample?
• « Cyanobacterial and eutrophication history » of the water body
• We decided to focus on water bodies used for drinking water production,
bathing and irrigation
• The Agency Artois-Picardie choose some rivers and canals
• Sampling frequency: 1 or 2 times a month during July, August and
Septembrer (until November for the AG Agency)
• Inquiry to the DDASS and collectivities
RESULTS
•52,6 % of the samples (170 in total) did contain
potentially toxic cyanobacteria.
•37 % of the samples did contain toxins (MCYST)
•17 % of the samples did contain toxins at concentrations
higher than 1 µg/l
RESULTS (Summer 2000)
Water
Agency
Chla µg/l
(min; max)
% samples with
cyanobacteria
potentially
toxic
RM
0.5; 265
41.9
23
6.4
166
-
LB
7.9; 159000*
93
89
61.9
51.3
14106.7
RMC
2.2 ; 118.9
31.3
6
6
14
-
AG
0; 74
70
64
10.4
10.7
-
AP
0; 70
27
3.3
0
0.06
-
* scum
% toxic
samples
% samples
with [tox] > 1
µg/l
[Tox] max
µg/l
[Tox]
max
in a
bloom
µg/l
EXAMPLES OF BLOOMS.
How to identify them visually?
Bloom of Anabaena.
Niveau de risque modéré
4m
Wind
Accumulation
Accumulation = 100 fois
4 cm
Vent
Accumulation = 1000 fois
Lac : vue en plan
Profil du lac
Vent
Anabaena accumulating along the dam under the wind action
Green pellets of Microcystis
Cyanobacteria mud
Picture by Bi-Eau
Accumulation of cyanobacteria along the shore
Picture by Bi-Eau
FOLLOWING UP
2001: Monitoring of the water bodies that reprensented a risk in
2000. We worked only with 2 Water Agencies (LB and AG). For
the other ones, the toxins did not reprensent a priority (no
obligation to make controls).
2002: The Water Agency LB did stop the monitoring and we
decided to concentrate our work on our geographic region. New
study with the Water Agency Adour-Garonne. Monitoring of all
the toxins (all the microcystins, Anatoxin a, Anatoxin a(s),
saxitoxins and LPS).
2003: With the new reglementation about MCYST LR, the Water
Agencies consider that it is not their role anymore to do the
monitoring. The managers and the owners of the water bodies
should take it over. The Water Agencies also do not feel
responsible for the communication about cyanobacteria as it is
essentailly a health problem. However, they will have to integrate
the parameters « cyanobacteria and MCYST » in the context of
the monitoring of ecological quality of water bodies (SEQ Plan
d’EAU). The way to integrate these new parameters is presently
under discussion.
SCREENING OF TOXIC BLOOMS IN 23
WATER BASINS OF THE ADOURGARONNE BASIN.
Data 2000-2001-2002
Localization of the water bodies
Mas Chaban
Cognac
Lavaud
Tardoire
Neuvic
Moulin Pinard
Bort-les-Orgues
Couze
St-Etienne-de-Cantalès
Cabrerets
Charpal
Villeneuve/Lot
Parentis
Loubéjac
Roucarié
Fontbonne
Rassisse
St Ferréol
Roques
Choldocogagna
Cammazes
Drinking water
Bathing water
Some examples of the cyanobacteria we found
Filaments
Anabaena sp: microcystins, neurotoxins [anatoxin a, anatoxin a(s)] and LPS.
Microcystins and neurotoxins
Planktothrix agardhii
Oscillatoria sp
Aphanizomenon sp
Saxitoxins
Colonies: Gomphospaeria (non toxic?) and Microcystis
Microcystis wesembergii
Gomphosphaeria sp
M. flos-aquae
M. aeruginosa
Cyanobacteria occurrence: Bilan 2000-2001-2002
Mas Chaban
Cognac
Lavaud
Tardoire
Neuvic
Moulin Pinard
Bort-les-Orgues
Couze
St-Etienne-de-Cantalès
Cabrerets
Charpal
Villeneuve/Lot
Parentis
Loubéjac
Roucarié
Fontbonne
Rassisse
Legend
St Ferréol
Roques
Choldocogagna
Absence de cyanobactéries
Présence de cyanobactéries (> 10 % du plancton)
Cammazes
Cyanobactéries (> 90 % plancton)
Mas Chaban
Cognac
Lavaud
Tardoire
Microcystines occurrence 2000, 2001 et
2002
Neuvic
Moulin Pinard
Bort-les-Orgues
Couze
St-Etienne-de-Cantalès
Cabrerets
Charpal
Villeneuve/Lot
Parentis
Loubéjac
Roucarié
Fontbonne
Rassisse
Légende
St Ferréol
Roques
Choldocogagna
Absence de cyanobactéries
Présence de cyanobactéries sans toxines
Cammazes
Cyanobactéries et teneur en toxines < 1 µg/l
Cyanobactéries et teneur en toxines > 1
µg/l
Cyanobactéries et teneurs en toxines >
10 µg/l
AEP
Roucarié
14
14
12
12
10
2000
8
2001
6
2002
4
Alerte 1
2
concentration
m icrocystines (µg/l)
concentration
m icrocystines (µg/l)
Fontbonne
0
10
2000
8
2001
6
2002
4
Alerte 1
2
0
août
septembre
octobre
août
m ois
septembre
octobre
m ois
BAIGNADE
Parentis
14
14
12
12
10
8
2000
Niveau 1
2001
6
2002
4
2
0
concentration en
m icrocystines (µg/l)
concentration en
m icrocystines (µg/l)
Lavaud
10
8
Niveau 1
2000
2001
6
2002
4
2
< 0.1 µg/l
0
août
octobre
m ois
août
octobre
m ois
Concentrations en microcystines totales d’août à octobre dans les 4 stations suivies de 2000 à 2002
PROFILS HPLC STANDARDS MICROCYSTINES
LR
RR
C
Max d’absorption:
239 nm
HPLC profile of a lake used for bathing
Bloom of Microcystis (different species). Presence of numerous
microcystin variants (YR, 1023-RR, 980-LR + 1030-YR, identified by
mass spectrometry).
Use of molecular tools to detect the toxic potential
M
1
2
3
4
5
6
M
Résultats amplification par PCR de la région mcyA. M : marqueurs de taille ; 1 : CHO 10 ;
2 : MC 08 ; 3 : BLO 07 ; 4 : P08 ; 5 : PCC7806 ; 6 : H2O.
The other toxins
•No neurotoxins [Anatoxin a, anatoxin a(s) and saxitoxins ] found.
So far, none neurotoxins found in France.
LPS found (difficult to separate the bacterial LPS from the cyanobacterial
ones). This Summer, high concentrations in a water body used as row water
for drinking water production.
No microcystins,
no neurotoxins
but LPS
31.8 ng/ml in row water. Lack of sanitary references. Réf: In Filand,
fever outbreak after consumption of a tap water containing 40 ng/ml LPS
which corresponded to LPS concentrations ranging from 200 à 1000 ng/ml in
row water.
Did we fulfill our objectives?
• Complete the EFFLOCYA data. Especially data missing in the South.
We have now data at a national level.
•Identify and list the water bodies which represent a risk and that
should be monitored regularly.
The sites are identified and the regular monitoring has still to
be done (the new reglementation will reinforce and give more power
to our advisies)
•To test and optimize the protocoles.
•To think about the possibilities of a long-term monitoring (sampling
frequency, with which actors, who is going to finance?…).
•To make the water managers and water users aware of this
environment and health problem.
•To think about the way to communicate the results (to inform
without causing panic). Most difficult part. The communication is not
working very well! It should change with the new reglementation as it
is specified that the public should be informed by panels in case of
health risk.
ANALYSES ASKED IN THE NEW REGLEMENTATION
STEP I: Observation under the microscope: Présence or absence of
cyanobactéria?
If presence of cyanobacteria
STEP II: Biomass: Chla and counting Î risk level according to the OMS
recommandations
In our lab, we add a step here:
measurement of the toxic
potention by PCR. If negative,
steps III and IV should not be
necessary.
STEP III: Research of microcystins with PP2a test
If test +
STEP IV: confirmation and identification of microcystines with HPLC or HPLC-MS
Monitoring in other European countries
• Monitoring very active in the Land
of Berlin
• Straight collaboration between the
Institute for Water, Air and Soil
Hygiene
(German
Federal
Environmental Agency which is the
representative of WHO in Germany)
and the numerous Water Agencies
(more than one hundred). Sampling:
samples concentrated on plankton net
and row water
• No reglementation but information
of the public by press articles and
panels at the lake’s shore with a
message such as « If you go in the
water until your knee’s level and you
do not see your feet anymore, there
is a risk of cyanobacteria bloom »
Portugal
• Straight collaboration between the Ministry for the
Health, the Ministry for the Environment and the
scientists
• Monitoring at a national level. Before starting the
monitoring, they spent one year standardizing the
methods (sampling, analyses…)
• Information of the public and the managers through a
nice folder
United-Kingdom
• Information of the doctors, pharmacist and veterinaries