Poisoning of Cattle Feeding on Allium ampeloprasum

Transcription

El-Sayed et al., J Vet Sci Med Diagn 2015, 4:4
http://dx.doi.org/10.4172/2325-9590.1000170
Journal of
Veterinary Science &
Medical Diagnosis
Case Report
a SciTechnol journal
Poisoning of Cattle Feeding on
Allium ampeloprasum (Egyptian
kurrat)
El-Sayed YS1*, El-Okle OSM2, Hassan SMH3 and Bakir NMA4
Abstract
Ten days after ingesting a large quantity of Allium ampeloprasum
(kurrat), 10 one-year-old calves and 2 eight-year-old cows in a
group of cattle were referred for voiding dark-red urine, generalized
jaundice, anemic or icteric mucous membranes, lack of appetite,
and lethargy. Hematologically, Heinz bodies – hemolytic anemia,
polychromasia, anisocytosis and leukocytosis were detected.
Biochemically, higher serum transaminases activities and bilirubin,
urea and uric acid levels were detected. As well as, an induced
lipid peroxidation and a perturbed antioxidant system; decreased
glutathione level, and glutathione reductase and catalase
activities were significantly detected. The treatment regimen of the
poisoned animals included the parenteral administration of fluids,
phosphorous and vitamins, and recovery occurred within one week
after initiating the treatment. This is the first report describing Allium
ampeloprasum toxicity in cattle with special reference to oxidative
status.
Keywords
Allium ampeloprasum;
peroxidation
Cattle;
Anemia;
Antioxidants;
Lipid
Introduction
Poisonous plants cause large economic losses to the livestock
industries throughout the world. This cost considered death losses and
specific reproductive losses in cattle and sheep. Fewer obvious costs
such as lost grazing opportunities, additional feed costs, increased
health care costs, management changes, increased culling costs and
lost weight gains [1]. The genus Allium includes onion, garlic, leek,
kurrat, chives, shallots and scallions were previously classified in the
family Alliaceae [2]. New classification of Angiosperm Phylogeny
Group, Alliaceae is now the subfamily Allioideae of the family
Amaryllidaceae [3]. Some of Allium species such as Allium cepa,
Allium sativum, Allium ampeloprasum and Allium schoenoprasum
are of toxicological importance as it contains toxic components
that may damage red blood cells and provoke hemolytic anemia
accompanied by the formation of Heinz bodies in the erythrocytes
of animals [4]. Cats, dogs, and cattle appear to be most susceptible to
the onion’s toxic effects, whereas sheep and goats appear to be most
resistant. This difference in susceptibility between species may be
due to differences in hemoglobin structure, as well as differences in
*Corresponding author: Yasser Said El-Sayed, Department of Veterinary
Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Damanhour
University, Damanhour 22511, Egypt, Tel: +20-100-108-5567; E-mail: yasser_
tf@yahoo.com, elsayed-ys@vetmed.dmu.edu.eg
Received: August 07, 2015 Accepted: September 04, 2015 Published:
September 08, 2015
International Publisher of Science,
Technology and Medicine
metabolism and detoxification within the gastrointestinal tract and
internal tissue organs [5].
N-propyl disulfide and sodium n-propylthiosulfate are the
main organo sulfur compounds [20]. The clinical signs observed
in cattle with onion toxicosis include off food, staggering, abortion
[5,16]. Although body temperatures are usually normal to low,
there may be fever up to 40.5°C. Death is common unless effective
treatment is provided, and surviving animals require a long period
of convalescence [6]. There is no specific antidote for onion toxicosis
but the treatment depends on whole blood transfusion if necessary,
intravenous fluids to control shock and dehydration. An empirical
therapy to limit oxidant-induced damage of erythrocytes is ascorbic
acid [7]. The main aim of this case report is to increase the attention of
veterinarians to the possibility of facing similar cases and to prepare
them for correct diagnosis and treatment, in addition to detect
biochemical alteration and status of oxidative stress in the blood of
affected animals.
Case Presentation
History and clinical observations
In April 2015, on a property at El-Bahera Governorate, Egypt,
10 one-year-old calves and 2 eight-year old cows clinically affected
in a group of 20 cattle (Bos taurus) with calves, commencing after
they had been feeding kurrat for consecutive 10 days. Clinically, the
affected animals exhibited signs of weakness, loss of appetite, dyspnea,
lethargy, tensmus, frequent voiding of dark red urine (Figure 1), pale
or icteric skin and mucus membranes (Figure 2), garlic odor on
breath, and a lowered body temperature (37.2-37.6°C) with exception
of one feverish cow (41.2°C). Samples of the kurrat fed to the cattle
were collected and submitted for definitive botanical identification
by an agronomist at the Herbarium Department in the Faculty of
Agriculture, Alexandria University. The results revealed that the
plant samples belonged to the family Alliaceae, and in particular, to
the cultivated vegetable A. ampeloprasum; Egyptian kurrat (Figure 3).
Hematological and serum biochemical assays
Venous blood samples were collected from poisoned animals (ills,
Figure 1: (A) Normal color urine of a control cow. (B) Dark-red colored urine
of a poisoned 1.5 years-old cow and (C) of a poisoned 1.5 years-old bull.
All articles published in Journal of Veterinary Science & Medical Diagnosis are the property of SciTechnol, and is protected by
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Citation: El-Sayed YS, El-Okle OSM, Hassan SMH, Bakir NMA (2015) Poisoning of Cattle Feeding on Allium ampeloprasum (Egyptian kurrat). J Vet Sci Med
Diagn 4:4.
doi:http://dx.doi.org/10.4172/2325-9590.1000170
and utilized for spectrophotometric assays of blood malondialdehyde
(MDA) [8] and GSH levels [9], and glutathione peroxidase (GPx) [10],
catalase (CAT) [11], and glutathione reductase (GR) [12] enzymatic
activities. Relative to the sera of the controls, the analytical findings
revealed that the poisoned animals exhibited elevated MDA (65.06 ±
4.28 nmol/L), bio-indicator of lipid peroxidation. These animals also
had significantly decreased GSH (1.35 ± 0.47 μmol/L) levels and GR
(538.8 ± 37.2 U/mg protein) and CAT (113.1 ± 7.8 U/mg protein)
enzymatic activities; these results are consistent with oxidative stressinduced hemolytic anemia. However, GPx activity (404.5 ± 20.5 U/
mg proteins) was significantly enhanced in the blood of poisoned
animals.
Figure 2: Heifer cow showing icteric skin and vulval mucous membrane.
Statistical analysis
All data are expressed as the mean ± standard deviations (SDs),
and the levels of significance are cited. The SPSS statistical package
version 17.0 for Windows (IBM, Armonk, NY, USA) was used for
all data analyses. Differences in values were analyzed by t-test.
Differences were deemed significant for p<0.05.
Table 1: Hematological indices in cattle ingested kurrat (A. ampeloprasum), The
mean values indicated by asterisk are significantly different between groups
(*p<0.05, **p<0.01). RBCs, red blood cells; Hb, hemoglobin; PCV, packed cell
volume; MCV, mean corpuscles volume; MCH, mean corpuscles hemoglobin;
- MCHC, mean corpuscles hemoglobin concentration; WBCs, white blood cells.
Group
Control
Intoxicated
Reference values
7 ± 1.2
2.8 ± 0.48**
5.0-10.0
Hb (g/dL)
12 ± 0.20
6.1 ± 1.4*
8-15
PCV (%)
32 ± 2.5
14.3 ± 2.9*
24-46
RBCs (106/µL)
Figure3: Allium ampeloprasum (Egyptian kurrat).
n=12) and non-poisoned healthy animals (controls, n=6) into plain
tubes with or without anticoagulant for routine hematological and
serum biochemical screening. Compared with the control animals, the
poisoned animals exhibited marked microcytic hypochromic anemia,
with leukocytosis, neutrophilia and lymphopenia. The microcytic
hypochromic hemolytic anemia was indicated by a decreased
red blood cells (RBCs) count (2.8 ± 0.48 ×106/µL), hemoglobin
concentration (Hb) (6.1 ± 1.4 g%), packed cell volume (PCV) (14.3
± 2.9%), mean corpuscular hemoglobin (MCH) (23.3 ± 3.2 pg)
and mean corpuscular hemoglobin concentration (MCHC) (42.7 ±
2.5%), which were below the normal ranges (Table 1). Polychromasia,
anisocytosis and Heinz bodies, in addition to a moderate increase in
neutrophils (50 ± 1.5%) and a decrease in eosinophils (6 ± 0.43%),
were consistent findings in the poisoned animals. With respect
to serum biochemistry, the poisoned animals exhibited increased
transaminases activity, particularly alanine aminotransferase (ALT)
(37.2 ± 1.30 U/dL), and levels of bilirubin (11.2 ± 2.23 mg/dL), urea
(58.24 ± 1.42 mg/dL) and uric acid (3.88 ± 0.31 mg/dL), which were
above the normal ranges (Table 2); these findings are consistent with
hepatorenal failure. No consistent changes were observed in any of
the other analyzed biochemical parameters, which included total
protein, albumin and globulin concentrations.
Blood antioxidants and lipoperoxidation
To evaluate antioxidant status, heparinized blood samples were
collected from poisoned and control animals, and then centrifuged at
4,000 rpm for 15 min at 4°C in a cooling centrifuge. Sera were collected
Volume 4 • Issue 4• 1000170
MCV (fL)
46 ± 5.4
54.6 ± 6.8
40-60
MCH (pg)
13 ± 2.4
23.3 ± 3.2*
11-17
MCHC (%)
33 ± 3.3
42.7 ± 2.5*
30-36
WBCs (103/µL)
6.4 ± 1.3
12.9 ± 4.3
4.0-12.0
Differential percentage (%)
Lymphocytes
58 ± 4.5
40 ± 4.3
62–63
Neutrophils
28 ± 5.4
50 ± 1.5*
15-33
Eosinophils
10 ± 0.33
6 ± 0.43*
0-20
Monocytes
5 ± 0.5
4 ± 1.2
0-8
280 ± 66.5
253.2 ± 49.9
100-800
Platelets (103/µL)
Table 2: Blood biomarkers of hepatorenal function, lipid peroxidation and
antioxidant molecules in cattle ingested kurrat (A. ampeloprasum). The
mean values indicated by asterisks are significantly different between
groups (*p < 0.05, **p<0.01, ***P<0.0001). ALT, alanine aminotransferase; AST,
aspartate aminotransferase; CAT, catalase; GPx, glutathione peroxidase; GR,
glutathione reductase; GSH, reduced glutathione; MDA, malondialdhyde.
Parameter
Control animals
Intoxicated
animals
Reference
range
Total protein (mg/dL)
7.42 ± 0.19
7.28 ± 0.46
7.5-7.6
Albumin (mg/dL)
3.61 ± 0.31
3.46 ± 0.30
2.5-3.8
Globulin (mg/dL)
3.22 ± 0.31
3.82 ± 0.43
3.0-3.5
AST (U/L)
47.15 ± 3.12
137.4 ± 14.27**
60-125
ALT (U/L)
14.5 ± 0.33
37.2 ± 1.30*
6.9-35
Bilirubin (mg/dL)
0.5 ± 0.03
11.2 ± 2.23*
0-1.6
Urea (mg/dL)
16.2 ± 2.2
58.24 ± 1.42*
6-22
Uric acid (mg/dL)
0.8 ± 0.04
3.88 ± 0.31*
0.4-1.2
10.39 ± 1.43
65.06 ± 4.28**
MDA (nmol/L)
GSH (μmole/L)
7.51 ± 0.31
1.35 ± 0.47***
GPx (U/mg protein)
184.7 ± 26.1
404.5 ± 20.5**
GR (U/mg protein)
937.2 ± 64.7
538.8 ± 37.2**
CAT (U/mg protein)
369.5 ± 18.1
113.1 ± 7.8**
• Page 2 of 4 •
Diagn 4:4.
Discussion
Acknowledgment
Onion-induced hemolytic anemia has been already known, and
the current case has now documented a similar effect of another Allium
spp.; kurrat. Similar findings were previously reported in bovines as
well as in other animal species such as canines, equines and ovines that
were fed a large amount of other Allium spp. (onion or garlic) [13,14].
The toxicity of Allium spp. is attributed to their contents of disulfides,
n-propyl disulfate, and S-methyl and S-prop(en)yl cysteine sulfoxides
(SMCOs) derived from amino acids [15]. Anaerobic bacteria play a role
in increasing the toxicity of SMCOs by hydrolyzing such compounds
in the rumen to thiosulfonate, which is then further metabolized to
dipropyl disulfides and dipropenyl disulfides [16]. These disulfides are
responsible for oxidative damage to erythrocytes [17]. SMCOs cause
a marked decrease in the activity of G6PD, in turn cause an increase
in the formation of methemoglobin and Heinz body count, and
reduction of GSH concentration in the erythrocytes of the poisoned
animals [14,18,19]. In erythrocytes, the declined GSH content was
associated with elevated levels of hydrogen peroxide (H2O2) [20].
H2O2 and MDA oxidize the sulfhydryl groups of Hb, resulting in its
denaturation. In the described case, a significant increase in MDA
was observed after the ingestion of A. ampeloprasum, suggesting
the production and/or accumulation of ROS, particularly H2O2, due
to degradation by CAT and the reduction of GR [21]. Because of
oxidative damage, denatured Hb would precipitate, aggregate and
bind to the erythrocyte membrane [21], thereby increasing formation
of Heinz bodies. In addition, lipid peroxidation is closely related
to erythrocyte deformity. Hemolytic anemia resulting from Allium
toxicity may be due to intravascular or extravascular hemolysis
[13]. Intravascular hemolysis, as indicated by increased serum
bilirubin, may occur because erythrocytes containing Heinz bodies
have decreased deformability and can burst when passing through
sinusoids or small capillaries [22]. Furthermore, direct oxidative
damage to the erythrocyte cell membrane contributes to cell lysis
[23], wherever formation of Heinz bodies and eccentrocytes increases
erythrocyte fragility and extravascular hemolysis [24].
The authors thank Vet. Dr. Mosád Zaid (Directorate of Veterinary Medicine,
Damanhour, Egypt) for his invaluable assistance in consultation and for providing
descriptions for poisoned animals.
The recommended treatment for kurrat toxicosis includes
restriction of access to the plant and the parenteral administration
of excess fluids, phosphorus, and vitamins, particularly vitamin C for
at least five consecutive days. Animals treated in this way recovered
within a week after the initiation of treatment. An additional and
potentially preventive recommendation is the administration of
oral antibiotics, which can provide the benefit of reducing ruminal
anaerobic bacteria that promote the formation of certain oxidative
substances [25]. A potential alternative to the unrestricted ingestion
of kurrat by ruminants might be to crush these plants and mix them
with the feed at no greater than 25% of the entire diet [26]. The
use of kurrat to feed ruminants is also possible in the context of a
protein-rich diet. Dietary proteins are important for the synthesis of
enzymes and for ensuring the availability of cofactors required for
antioxidative reactions [27].
Conclusion
The consumption of A. ampeloprasum by ruminants resulted
in poisoning. Allium toxicosis is typically diagnosed through a
combination of history, clinical signs, and microscopic confirmation
of oxidative Heinz body – hemolytic anemia and disturbed hepatorenal
function. Restricting access to the Allium spp. and treating poisoned
animals with fluids, phosphorous, and vitamin supplements induced
successful recoveries.
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Top
Author Affiliations
Department of Veterinary Forensic Medicine and Toxicology, Faculty of
Veterinary Medicine, Damanhour University, Damanhour 22511, Egypt
2
Department of Veterinary Forensic Medicine and Toxicology, Faculty of
Veterinary Medicine, Alexandria University, Edfina 22576, Egypt
3
Department of Clinical Pathology, Alexandria Regional Lab. Animal
Health Research Institute, Alexandria, Egypt
4
Department of Animal Medicine and Infectious Diseases, Faculty of
Veterinary Medicine, Damanhour University, Damanhour 22511, Egypt
1
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