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PloS one2015; 10(9); e0136785; doi: 10.1371/journal.pone.0136785

Hypoglycin A Content in Blood and Urine Discriminates Horses with Atypical Myopathy from Clinically Normal Horses Grazing on the Same Pasture.

Abstract: Hypoglycin A (HGA) in seeds of Acer spp. is suspected to cause seasonal pasture myopathy in North America and equine atypical myopathy (AM) in Europe, fatal diseases in horses on pasture. In previous studies, this suspicion was substantiated by the correlation of seed HGA content with the concentrations of toxic metabolites in urine and serum (MCPA-conjugates) of affected horses. However, seed sampling was conducted after rather than during an outbreak of the disease. The aim of this study was to further confirm the causality between HGA occurrence and disease outbreak by seed sampling during an outbreak and the determination of i) HGA in seeds and of ii) HGA and MCPA-conjugates in urine and serum of diseased horses. Furthermore, cograzing healthy horses, which were present on AM affected pastures, were also investigated. AM-pastures in Germany were visited to identify seeds of Acer pseudoplatanus and serum (n = 8) as well as urine (n = 6) from a total of 16 diseased horses were analyzed for amino acid composition by LC-ESI-MS/MS, with a special focus on the content of HGA. Additionally, the content of its toxic metabolite was measured in its conjugated form in body fluids (UPLC-MS/MS). The seeds contained 1.7-319.8 μg HGA/g seed. The content of HGA in serum of affected horses ranged from 387.8-8493.8 μg/L (controls < 10 μg/L), and in urine from 143.8-926.4 μg/L (controls < 10 μg/L), respectively. Healthy cograzing horses on AM-pastures showed higher serum (108.8 ± 83.76 μg/L) and urine concentrations (26.9 ± 7.39 μg/L) compared to control horses, but lower concentrations compared to diseased horses. The range of MCPA-carnitine and creatinine concentrations found in diseased horses in serum and urine were 0.17-0.65 mmol/L (controls < 0.01), and 0.34-2.05 μmol/mmoL (controls < 0.001), respectively. MCPA-glycine levels in urine of cograzing horses were higher compared to controls. Thus, the causal link between HGA intoxication and disease outbreak could be further substantiated, and the early detection of HGA in cograzing horses, which are clinically normal, might be a promising step in prophylaxis.
Publication Date: 2015-09-17 PubMed ID: 26378918PubMed Central: PMC4574941DOI: 10.1371/journal.pone.0136785Google Scholar: Lookup
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Summary

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This research article investigates a substance called hypoglycin A (HGA) that is found in certain seeds and is thought to cause serious diseases in horses. Scientists have found an increase in this substance in horses that have become sick, suggesting it may trigger an outbreak of disease. This study was designed to examine this correlation more closely, by testing for the substance both in seeds and affected horses during an outbreak, and also in healthy horses grazing in the same area.

Methodology

  • Researchers conducted their study in Germany, where outbreaks of equine atypical myopathy (AM), a potentially fatal disease in horses, have been linked to HGA.
  • They visited pastures where outbreaks of AM had occurred and collected samples of Acer pseudoplatanus seeds, suspected to contain HGA.
  • Blood and urine samples were taken from 16 diseased horses and analyzed for their amino acid composition, with special focus placed on the presence of HGA. Additionally, its toxic metabolite MCPA was also detected in its conjugated form.
  • To compare, similar samples were obtained from healthy horses that were grazing on the same pastures.

Findings

  • The researchers discovered that the seeds contained HGA varying from 1.7 to 319.8 μg HGA/g seed.
  • The diseased horses presented high concentrations of HGA in their serum and urine, with the range found to be significantly higher than that in control horses (non-exposed).
  • Interestingly, the healthy horses that were present on the same pastures as the diseased ones also exhibited higher HGA levels in their blood and urine than control horses, though not as high as the diseased horses.
  • The concentrations of the toxic metabolites MCPA-carnitine and creatinine were also found to be significantly increased in the urine and serum of diseased horses.
  • Conversely, a higher level of MCPA-glycine was found in the urine of cograzing horses as compared to control horses.

Conclusion

  • The data collected strongly substantiates the causal link between HGA and the outbreak of diseases in horses, reinforcing the suspicion that Acer seeds are a source of HGA intoxication that can trigger serious equine diseases.
  • The study also suggests that even healthy horses grazing on the same pastures as diseased horses might be ingesting similar toxic substances, albeit at lower quantities.
  • The findings indicate that early detection of elevated HGA in horses that appear clinically normal might be a valuable step towards preventing outbreaks of the illness.

Cite This Article

APA
Bochnia M, Ziegler J, Sander J, Uhlig A, Schaefer S, Vollstedt S, Glatter M, Abel S, Recknagel S, Schusser GF, Wensch-Dorendorf M, Zeyner A. (2015). Hypoglycin A Content in Blood and Urine Discriminates Horses with Atypical Myopathy from Clinically Normal Horses Grazing on the Same Pasture. PLoS One, 10(9), e0136785. https://doi.org/10.1371/journal.pone.0136785

Publication

ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 10
Issue: 9
Pages: e0136785

Researcher Affiliations

Bochnia, M
  • Institute of Agricultural and Nutritional Sciences Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
Ziegler, J
  • Department of Molecular Signal Processing, Leibniz Institute of Plant Biochemistry, Halle (Saale), Germany.
Sander, J
  • Screening Labor, Hanover, Germany.
Uhlig, A
  • Department of Large Animals Medicine, Faculty of Veterinary Medicine, Leipzig, Germany.
Schaefer, S
  • Institute of Agricultural and Nutritional Sciences Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
Vollstedt, S
  • Traditional Chinese Equine Medicine, Bokholt-Hanredder, Germany.
Glatter, M
  • Institute of Agricultural and Nutritional Sciences Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
Abel, S
  • Department of Molecular Signal Processing, Leibniz Institute of Plant Biochemistry, Halle (Saale), Germany.
Recknagel, S
  • Department of Large Animals Medicine, Faculty of Veterinary Medicine, Leipzig, Germany.
Schusser, G F
  • Department of Large Animals Medicine, Faculty of Veterinary Medicine, Leipzig, Germany.
Wensch-Dorendorf, M
  • Institute of Agricultural and Nutritional Sciences Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
Zeyner, A
  • Institute of Agricultural and Nutritional Sciences Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.

MeSH Terms

  • Acer / poisoning
  • Animals
  • Carnitine / analogs & derivatives
  • Carnitine / blood
  • Carnitine / urine
  • Disease Outbreaks
  • Horse Diseases / blood
  • Horse Diseases / diagnosis
  • Horse Diseases / urine
  • Horses
  • Hypoglycins / blood
  • Hypoglycins / poisoning
  • Hypoglycins / urine
  • Muscular Diseases / blood
  • Muscular Diseases / chemically induced
  • Muscular Diseases / veterinary
  • Plant Poisoning / blood
  • Plant Poisoning / urine
  • Plant Poisoning / veterinary
  • Plants, Toxic / poisoning
  • Seeds / chemistry
  • Seeds / poisoning
  • Tandem Mass Spectrometry

Conflict of Interest Statement

Competing Interests: The authors have declared that no competing interest exist.

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