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Jan Marczuk, Krzysztof Lutnicki, Maciej Gaj^cki*©
Chair and Clinic of Internal Medicine, Department of Internal Diseases of Farm Animals and Horses, University of Live Sciences in Lublin, 20-612 Lublin, Poland
email: doktorjm@o2.pl *Department of Veterinary Prophylasis and Feed Hygiene, Faculty of Veterinary Medicine University of Warmia andMazury in Olsztyn, 10-718 Olsztyn, Poland
CHRONIC FUSARIUM-DERIVED MYCOTOXINS INTOXICATION IN DAIRY COWS HERD - CASE STUDY
Abstract.
Mycotoxin intoxications from economical and toxicological points of view become recently more and more important. In clinical practice they are rarely diagnosed due to the lack of characteristic clinical signs of illness. This paper concerns the clinical case of chronic intoxication with mycotoxins derived from Fusarium (deoxynivalenol- DON, zearalenon-ZEA), which were the reason for cows' death in a herd in Eastern Poland , where the high levels of DON and ZEA were estimated in affected cows' serum and as well as simultaneousl abnormal hematological parameters.
Key words: mycotoxins, mycotoxicoses, zearalenon, deoxynivalenol, dairy
cattle
Introduction
Mycotoxins are the secondary metabolites originating from some fungi existing on the plants during their vegetation or produced at the time of their storage. The favorable factors are the inappropriate corn storage conditions (high humidity and temperature with air access) of the plants which were contaminated during the growth phase. In a contaminated part of fodder fungi (mycoses) often exist as focal spots , which is the additional difficulty in mycotoxins detection. Moreover contemporary methods and technology of cows feeding promote equal mixing and incorporation into the whole feed dose of the contaminated fodder elements, which leads to the diminished fodder quality. The mycotoxins intoxications must be the predominant suspicion in all cases of sudden low milk production, as well as initial agent of production effects diminishing and enhanced mortality in the herd. Due to the immunosuppression, which is evoked by mycotoxins the course of illnesses is different and usually unspecific. There are usually no characteristic pathological signs during section examinations, except the generalized inflammation of enteric system mucosa and nonspecific, irritant changes in the other mucosal membranes. These signs do not allow to decide about the certain diagnosis. Besides, the change of fodder, elimination of suspicious (or contaminated) parts is impossible. In this instance very useful may be the proof which is based on adding to contaminated fodder the preparations which can chemisorb the mycotoxins or adding the uncontaminated fodder and in this way diminishing the concentration of mycotoxin. The observation of the effects of these actions can allow to make a diagnosis ex juvantibus. In contrast to monogastric animals the ruminants are more resistant to
© Jan Marczuk, Krzysztof Lutnicki, Maciej Gaj^cki, 2009
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mycotoxins intoxication due to their ability to mycotoxins degradation in the rumen, reticulum and omasum. Nevertheless, the lactating cows are more susceptible to intoxication in comparison to beef ones- due to the production stress. The most common and potentially toxic for dairy cows mycoses and mycotoxins according Whitlow and Hagler (Whitlow and Hagler 2005) are presented in Table 1.
Tab.1.
Mycoses (fungi genus) Mycotoxins
Aspergillus Aflatoxin, Ochratoxin, Sterigmatocytsin, Fumitremorgens, Fumigaclavines, Fumitoxins, Cyclopiazonoic acid, Gliotoxin
Fusarium Deoksynivalenol, Zearalenone, T-2 Toxin, Fumonizin, Moniliformin, Nivalenol, Diacetoxyscirpenol, Butenolide, Neosolaniol, Fusaric acid, Fusaproliferin, Fusarin C
Penicillium Ochratoxin, PR Toxin, Patulin, Penicillic acid, Citrinin, Penetrem, Cyklopiazonic acid, Roquefortine, Isofumigaclavines A and B, Mycophenolic acid
Claviceps Ergot alkaloids in seed/grain of small grains, sorghum, grasses
Epichloe and Neotyphodium Ergot alkaloids in fescue grass
Stachybotrys Stachybotryotoxins, trichothecenes
The influence of mycotoxins on cows organism is show below in Fig. 1.
* Gastroenteritis
* ltn9mr h6ati . Intettiul himGnftages
- Lflw coiiMptiori rttta , fcnp,,^ ,umen fvnctic.n
* 0tfar1an . Mt,tf№
* Brtwnfc i™ . Ketosii.
Ryc. 1. The main consequences of mycotoxins intoxications in cattle. (according Inês Rodrigues, Product Manager (Mycofix® Plus product line) - Biomin
GmbH)
The main consequences of intoxication are :
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- the lack or diminishing of appetite
- lower incorporation of food components and inhibition of their metabolism
- hormonal and immunological dysfunction
- deregulation of rumen microenvironment (especially in saprophytic bacterium growth)
The certain diagnosis of mycotoxemia is difficult due to a great amount of potentially pathogenic mycotoxins, their multidirectional influence on animals, heterogeneous clinical symptoms and lack of specific diagnostic methods. Additionally, there are obstructions in gathering a representative proof of contaminated fodder. Mycotoxins intoxication can be suspected in all cases of illness with atypical course, which are resistant to classic therapy.
Clinical case report.
In the dairy cows herd consisting of 90 animals in February 2008 existed accidental cows mortality, including 10 lactating cows and 2 heifers. The sick animals did not exhibit any signs of illness until their death. The appetite and thirst were normal, only in part of the cows occurred salivation just after the TMR digestion. Only the sick animals had problems with motion, they had big problems with normal displacement in the stable (barn). Just before death they were in lying position.
The feeding was based on hay silage, maize silage, soybean and grain , CCM prefix and mineral supplements. Besides TMR there was free access to the fullportion fodder station containing 21% of protein. The mean milk yield in the herd during 305 days of lactation was 8000-11830 kg.
Material and methods.
Samples.
The material for hematological and biochemical estimations were blood serum samples collected from nine cows with clinical signs of the disease.
Mycotoxins assays.
ZEA determinations were made with HPLC-FLD method (Zearala TestWB, Instruction Manual, VICAM) and DON with HPLC-UV method (DonTestWB, Instruction Manual, VICAM).
In the blood samples there were the basic hematological (WBC, RBC, Hb, Ht, neutrophils, eozynphils, lymphocyte, monocyte, platelet count, total protein, albumins, globulins, AST, Na, K, urea, total Ca, Ca++, P, Mg, fibrinogen and protrombin time) determined by kinetic method with EPOLL-20 analyzer and Mindray chemistry analyzer BS -130.
Results
Obtained results of DON and ZEA concentration in blood serum are presented in Table.1.
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Tab. 1. The concentration of investigated mycotoxins.
Deoxynivalenol (DON) Zearalenone (ZEA)
No. of cow concentration in serum concentration in serum
[ng/mL] [ng/mL]
1 16,31 4,60
2 23,12 7,34
3 12,76 negative
8 8,76 negative
9 5,10 3,2
Reference values < 5 < 5
The mean DON concentrations were in all investigated animals higher than normal in ranges 5,10 - 23,12 ng/mL , 2 cows were free from ZEA toxins, only in one cow ZEA concentration was higher than normal. In 8 of 9 investigated animals the higher number of leucocytes were determined, in range 13,88 do 53,22 x 109 /L with shift to the right (segmented neutrophil amount 43-74%). In 6 cows simultaneously was observed the marked lymphocytopenia (18-44%) and increased amount of platelets (8,33 - 17,85 x 105 /^L). In all affected animals were the increased Ht and Na concentration determined. Biochemical analysis shows the increased AST activity in range 105 U/L do 2 034U/L , high concentration of total protein in range 71 g/L do 106 g/L and fibrinogen over 6,95g/L, associated with decreased total protein/fibrinogen ratio. Botulism was excluded microbiologically. Necropsy examination: pulmonary edema, foam exudates in bronchial tract, epicardial extravasations, submucosal extravasations in the gut and its inflammation.
Discussion.
Nowadays the mycotoxins are one of the pivotal primary agents of economical failures in dairy cows farms. The expense of examinations and their difficulty make them uncommon for many farmers. According to Driehuis et al. investigations the intensity of mycotoxins appearance in fodder proofs on milk farms is about 81% DON and 46% ZEA. Nevertheless, the experimental investigations based on clinical observations and treatment effects in animals administered with only one toxin give no information about the clinical course of illness in field circumstances and have a minimal importance. Ruminants have a physiological ability to diminish the toxic influence of mycotoxins. Ochratoxin OTA which is produced by Penicillum in the rumen is reduced to lower toxic compounds - ochratoxin alfa and phenylalanine. On the other hand, in the case of ZEA it comes to conversion into alfa zearalenon, which exhibits stronger estrogenic action.
Deoxynivalenol (DON) is commonly present in fodder, named Vomitoxin, because of its vomiting properties in affected swine. The results of experimental investigation carried out in milk cows suggest that DON ingestion leads to appetite lack or diminishing, gastroenteric ulcerations, especially in the rumen and reticulum mucosa, milk yield diminishing, increase in milk somatic cells count, fertility
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disturbances. Moreover DON caused rumen fermentation disturbances and diminished the amount of digestible protein, which is transported into duodenum.
Zearalenon (ZEA) - has a chemical structure like the natural estrogens. Intoxication leads to disturbances in reproduction system: estrus signs in immature cows, irregular periods in estrus cycle, silent estrus, fetus death, abortions, placental retention, uterus and mammary gland inflammation. Besides during ZEA intoxications appears the lack of appetite, decreased milk yield, enteritis, diarrhea. In contrast to our results, Korostelev i wsp., during the experimental investigations carried out on intoxicated cows observed no hematological changes. Many others investigators showed the increased concentrations of total protein and globulin after 42 days in experimental intoxication with Fusarium toxins. The experiments carried out on mice administered with DON feed exhibited significant decrease in globulin concentration and increase in albumin one. Increased level of total protein may be the consequence of dehydratation or fluid sequestration, which was manifested by higher Ht, increased RBC level and Na concentration. Besides, this increase in total protein level, which was observed by many investigators, may be a consequence of acute or chronic inflammation and increased fibrinogen level associated with diminishing plasma protein/fibrinogen ratio. The increased AST activity suggests the hepatic insufficiency in the course of Fusarium mycotoxins intoxication. These mycotoxins may have a direct influence on some hepatic globulin synthesis. The results of our investigation of cows with increased AST activity exhibit a significant lower globulin concentration, beyond the reference values. DON as well as ZEA intoxication lead to enteritis.
Conclusions:
1. Fusarium toxins intoxication in dairy cows breeding leads to hematological and biochemical disturbances.
2. Chronic mycotoxicosis (DON, ZEA) may evoke no clinical symptoms.
3. Significant clinical symptoms and lack of treatment usually cause the death of animals in consequence of heart pulmonary insufficiency (oedema pulmonum).
4. Chronic mycotoxicosis (DON, ZEA) leads to hepatic injury and decreased immunity.
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