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      In vitro biodegradation of cyanotoxins in the rumen fluid of cattle

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          Abstract

          Background

          In countries around the Baltic Sea grazing ruminants have access to and drink, surface water from lakes, rivers and in several coastal regions. The water quality of these naturally occurring reservoirs affects performance and health of livestock. In the Baltic Sea both microcystin (MC) and nodularin (NOD) occurs as cyclic peptides and have hepatotoxic effects. Although cattle obviously have died after consuming contaminated water very little information is available as to how susceptible ruminants are to the toxins produced by cyanobacteria. The critical question as to whether the rumen microflora might constitute a protective shield is unresolved. For this reason our aim is to investigate a possible degradation rate of these toxins in rumen.

          Results

          The ability of rumen microorganisms to degrade certain important cyanotoxins (MC-LR, YR, RR and NOD) was studied in vitro by incubating with rumen fluid at three different concentrations (0.05, 0.5 and 5 μg/mL) for 3 h. The degradation efficiencies were determined by LC-MS (ESI) positive mode. Degradation was observed in the following order MC-RR 36%, NOD 35%, MC-RR 25% and MC-LR 8.9% at lower concentrations within 3 h. However, average degradation was observed at concentration of 0.5 μg/mL. No degradation was observed in higher concentrations for entire 3 h. The present results reveal that the degradation was both dose and time dependent.

          Conclusions

          In conclusion the present results suggest that the rumen microbial flora may protect ruminants from being intoxicated by Cyanotoxins.

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          Most cited references31

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          The toxicology of microcystins.

          Microcystins are a family of more than 50 structurally similar hepatotoxins produced by species of freshwater cyanobacteria, primarily Microcystis aeruginosa. They are monocyclic heptapeptides, characterised by some invariant amino acids, including one of unusual structure which is essential for expression of toxicity. Microcystins are chemically stable, but suffer biodegradation in reservoir waters. The most common member of the family, microcystin-LR (L and R identifying the 2 variable amino acids, in this case leucine and arginine respectively) has an LD50 in mice and rats of 36-122 microg/kg by various routes, including aerosol inhalation. Although human illnesses attributed to microcystins include gastroenteritis and allergic/irritation reactions, the primary target of the toxin is the liver, where disruption of the cytoskeleton, consequent on inhibition of protein phosphatases 1 and 2A, causes massive hepatic haemorrhage. Microcystins are tight-binding inhibitors of these protein phosphatases, with inhibition constants in the nanomolar range or lower. Uptake of microcystins into the liver occurs via a carrier-mediated transport system, and several inhibitors of uptake can antagonise the toxic effects of microcystins. The most effective of these is the antibiotic rifampin (a drug approved for clinical use), which protects mice and rats against microcystin-induced lethality when given prophylactically and, in some cases, therapeutically.
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            Microcystin-producing blooms--a serious global public health issue.

            The investigation on microcystin topics is increasing due to the related ecological and public health risks. Recent investigation confirms a gap in establishing global patterns relating a particular environment to the bloom occurrence of a species and the production of certain microcystin variants. All the results concerning the environmental effects on the microcystin synthesis of one species must be checked in the light of genome diversity. Thus, the poisoning risks of a bloom depend on the strain causing toxicity. To be more effective, specific water treatment methods are required for blooms of different microcystin producing species (such as colonial and filamentous cyanobacteria found in stratified and unstratified water bodies, respectively). With the increasing number of new microcystin variants discovered, the development of new rapid, inexpensive and sensitive enough monitoring methods to promptly screen simultaneously a great diversity of toxins and also check their toxic effects is becoming necessary.
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              First identification of the hepatotoxic microcystins in the serum of a chronically exposed human population together with indication of hepatocellular damage.

              Hepatotoxic microcystins (MCs) are the most commonly reported cyanotoxins in eutrophic freshwaters. In 1996, human intoxications by MCs caused deaths of 76 patients at Caruaru dialysis centers in Brazil. So far, there have been no direct evidences of MC occurrence in human tissue in consequence of exposure to MC. In this study, we improved cleanup procedures for detecting MCs in serum sample using liquid chromatography-mass spectrometry, and confirmed for the first time the presence of MCs in serum samples (average 0.39 ng/ml, which amounts to ca. 1/87 of the concentrations found in tissue samples of the Caruaru victims) of fishermen at Lake Chaohu. Daily intake by the fishermen was estimated to be in the range of 2.2-3.9 microg MC-LReq, whereas the provisional World Health Organization tolerable daily intake (TDI) for daily lifetime exposure is 0.04 microg/kg or 2-3 microg per person. Moreover, statistical analysis showed closer positive relationships between MC serum concentrations and concentrations of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase than between the MC concentrations and other biochemical indicators. Thus, the data raise the question whether extended exposure in the range of the TDI or up to a factor of 10 above it may already lead to indication of liver damage. The results also demonstrate a risk of health effects from chronic exposure to MCs at least for populations with high levels of exposure, like these fishermen.
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                Author and article information

                Contributors
                Journal
                BMC Vet Res
                BMC Vet. Res
                BMC Veterinary Research
                BioMed Central
                1746-6148
                2014
                8 May 2014
                : 10
                : 110
                Affiliations
                [1 ]Department of Anatomy, Physiology and Biochemistry, Faculty of Veterinary Medicine and Animal Science, SLU, Box 7011 750 07 Uppsala, Sweden
                [2 ]Department of Food Science, Swedish University of Agricultural Sciences, SLU 750 07 Uppsala, Sweden
                Article
                1746-6148-10-110
                10.1186/1746-6148-10-110
                4018535
                962acff6-5918-4c12-b64a-89bf0a857862
                Copyright © 2014 Manubolu et al.; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.

                History
                : 22 May 2013
                : 1 May 2014
                Categories
                Research Article

                Veterinary medicine
                biodegradation,microcystins,nodularin,rumen microbial flora
                Veterinary medicine
                biodegradation, microcystins, nodularin, rumen microbial flora

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