FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / BMC Vet Res / Biomarkers for ragwort poisoning in horses: identification o...
BMC veterinary research2008; 4; 30; doi: 10.1186/1746-6148-4-30

Biomarkers for ragwort poisoning in horses: identification of protein targets.

Abstract: Ingestion of the poisonous weed ragwort (Senecio jacobea) by horses leads to irreversible liver damage. The principal toxins of ragwort are the pyrrolizidine alkaloids that are rapidly metabolised to highly reactive and cytotoxic pyrroles, which can escape into the circulation and bind to proteins. In this study a non-invasive in vitro model system has been developed to investigate whether pyrrole toxins induce specific modifications of equine blood proteins that are detectable by proteomic methods. Results: One dimensional gel electrophoresis revealed a significant alteration in the equine plasma protein profile following pyrrole exposure and the formation of a high molecular weight protein aggregate. Using mass spectrometry and confirmation by western blotting the major components of this aggregate were identified as fibrinogen, serum albumin and transferrin. Conclusions: These findings demonstrate that pyrrolic metabolites can modify equine plasma proteins. The high molecular weight aggregate may result from extensive inter- and intra-molecular cross-linking of fibrinogen with the pyrrole. This model has the potential to form the basis of a novel proteomic strategy aimed at identifying surrogate protein biomarkers of ragwort exposure in horses and other livestock.
Get Full Text
Publication Date: 2008-08-08 PubMed ID: 18691403PubMed Central: PMC2527303DOI: 10.1186/1746-6148-4-30Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article describes a study in which a non-invasive system was developed to identify specific changes in horse blood proteins upon exposure to toxins from the ragwort weed. The results of the study suggest that a proteomic strategy can be developed to identify protein biomarkers of ragwort exposure in horses.

Objective of the Study

  • The objective of the study was to investigate whether toxins from ragwort, a poisonous weed, induce specific modifications in horse blood proteins. These modifications, if identifiable, could serve as biomarkers for ragwort poisoning in horses.

Methodology

  • An in vitro model was developed to emulate the impact of ragwort poisoning on horse blood proteins.
  • The key toxins in ragwort are pyrrolizidine alkaloids, which transform to highly reactive and toxic pyrroles. In this study, the effects of these pyrroles on the protein structure of horse plasma were analyzed using techniques such as gel electrophoresis, mass spectrometry, and western blotting.

Results of the Study

  • The study found that exposure to pyrrole toxins leads to significant changes in horse plasma proteins.
  • One of the key findings was the formation of a high-molecular-weight protein aggregate after pyrrole exposure.
  • Further analysis identified the primary components of this aggregate as fibrinogen, serum albumin, and transferrin.

Conclusions of the Study

  • The study confirmed that exposure to pyrrolic metabolites leads to modifications in equine plasma proteins.
  • The high-molecular-weight aggregate may form as a result of extensive cross-linking of fibrinogen with the pyrrole toxin.
  • These findings open the path for developing a proteomic strategy designed at identifying surrogate protein biomarkers, which can prove helpful in the early detection of ragwort exposure in horses and other livestock.

Cite This Article

APA
Moore RE, Knottenbelt D, Matthews JB, Beynon RJ, Whitfield PD. (2008). Biomarkers for ragwort poisoning in horses: identification of protein targets. BMC Vet Res, 4, 30. https://doi.org/10.1186/1746-6148-4-30

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 4
Pages: 30

Researcher Affiliations

Moore, Rowan E
  • Proteomics and Functional Genomics Research Group, Faculty of Veterinary Science, University of Liverpool, Crown Street, Liverpool, L69 7ZJ, UK. r.e.moore@liverpool.ac.uk
Knottenbelt, Derek
    Matthews, Jacqueline B
      Beynon, Robert J
        Whitfield, Phillip D

          MeSH Terms

          • Animals
          • Biomarkers / metabolism
          • Blood Proteins / metabolism
          • Blotting, Western
          • Fibrinogen / metabolism
          • Hemoglobins / metabolism
          • Horse Diseases / diagnosis
          • Horse Diseases / etiology
          • Horses / blood
          • Horses / metabolism
          • Plant Poisoning / diagnosis
          • Plant Poisoning / etiology
          • Plant Poisoning / veterinary
          • Plants, Toxic
          • Proteomics
          • Pyrroles / poisoning
          • Pyrrolizidine Alkaloids / poisoning
          • Senecio / poisoning
          • Time Factors

          References

          This article includes 30 references
          1. Mendel VE, Witt MR, Gitchell BS, Gribble DN, Rogers QR, Segall HJ, Knight HD. Pyrrolizidine alkaloid-induced liver disease in horses: an early diagnosis.. Am J Vet Res 1988;49:572–578.
            pubmed: 3377320
          2. West HJ. Clinical and pathological studies in horses with hepatic disease.. Equine Vet J 1996;28:146–156.
            pubmed: 8706647
          3. Mattocks AR. Chemistry and toxicology of pyrrolizidine alkaloids.. London , Academic Press; 1986.
            pubmed: 0
          4. Cheeke PR. Toxicity and metabolism of pyrrolizidine alkaloids.. J Anim Sci 1988;66:2343–2350.
            pubmed: 3049495
          5. Stegelmeier BL, Edgar JA, Colegate SM, Gardner DR, Schoch TK, Coulombe RA, Molyneux RJ. Pyrrolizidine alkaloid plants, metabolism and toxicity.. J Nat Toxins 1999;8:95–116.
            pubmed: 10091131
          6. Witte L, Ernst L, Adam H, Hartmann T. Chemotypes of two pyrrolizidine alkaloid-containing Senecio species.. Phytochemistry 1992;31:559–565.
            doi: 10.1016/0031-9422(92)90038-Rgoogle scholar: lookup
          7. Macel M, Vrieling K, Klinkhamer PG. Variation in pyrrolizidine alkaloid patterns of Senecio jacobaea.. Phytochemistry 2004;65:865–873.
            doi: 10.1016/j.phytochem.2004.02.009pubmed: 15081286google scholar: lookup
          8. Williams DE, Reed RL, Kedzierski B, Dannan GA, Guengerich FP, Buhler DR. Bioactivation and detoxication of the pyrrolizidine alkaloid senecionine by cytochrome P-450 enzymes in rat liver.. Drug Metab Dispos 1989;17:387–392.
            pubmed: 2571477
          9. Glowaz SL, Michnika M, Huxtable RJ. Detection of a reactive pyrrole in the hepatic metabolism of the pyrrolizidine alkaloid, monocrotaline.. Toxicol Appl Pharmacol 1992;115:168–173.
            doi: 10.1016/0041-008X(92)90320-Rpubmed: 1641851google scholar: lookup
          10. Reed RL, Ahern KG, Pearson GD, Buhler DR. Crosslinking of DNA by dehydroretronecine, a metabolite of pyrrolizidine alkaloids.. Carcinogenesis 1988;9:1355–1361.
            doi: 10.1093/carcin/9.8.1355pubmed: 3402032google scholar: lookup
          11. Kim HY, Stermitz FR, Molyneux RJ, Wilson DW, Taylor D, Coulombe RA. Structural influences on pyrrolizidine alkaloid-induced cytopathology.. Toxicol Appl Pharmacol 1993;122:61–69.
            doi: 10.1006/taap.1993.1172pubmed: 8378933google scholar: lookup
          12. Estep JE, Lame MW, Morin D, Jones AD, Wilson DW, Segall HJ. [14C]monocrotaline kinetics and metabolism in the rat.. Drug Metab Dispos 1991;19:135–139.
            pubmed: 1673386
          13. Pan LC, Lame MW, Morin D, Wilson DW, Segall HJ. Red blood cells augment transport of reactive metabolites of monocrotaline from liver to lung in isolated and tandem liver and lung preparations.. Toxicol Appl Pharmacol 1991;110:336–346.
            doi: 10.1016/S0041-008X(05)80016-Xpubmed: 1909819google scholar: lookup
          14. Mattocks AR, Jukes R. Recovery of the pyrrolic nucleus of pyrrolizidine alkaloid metabolites from sulphur conjugates in tissues and body fluids.. Chem Biol Interact 1990;75:225–239.
            doi: 10.1016/0009-2797(90)90120-Cpubmed: 2369787google scholar: lookup
          15. Mattocks AR, Jukes R. Detection of sulphur-conjugated pyrrolic metabolites in blood and fresh or fixed liver tissue from rats given a variety of toxic pyrrolizidine alkaloids.. Toxicol Lett 1992;63:47–55.
            doi: 10.1016/0378-4274(92)90106-Tpubmed: 1412522google scholar: lookup
          16. Seawright AA, Hrdlicka J, Wright JD, Kerr DR, Mattocks AR, Jukes R. The identification of hepatotoxic pyrrolizidine alkaloid exposure in horses by the demonstration of sulphur-bound pyrrolic metabolites on their hemoglobin.. Vet Hum Toxicol 1991;33:286–287.
            pubmed: 1858316
          17. Seawright AA, Kelly WR, Hrdlicka J, McMahon P, Mattocks AR, Jukes R. Pyrrolizidine alkaloidosis in cattle due to Senecio species in Australia.. Vet Rec 1991;129:198–199.
            pubmed: 1957473
          18. Winter H, Seawright AA, Hrdlicka J, Mattocks AR, Jukes R, Wangdi K, Gurung KB. Pyrrolizidine alkaloid poisoning of yaks: diagnosis of pyrrolizidine alkaloid exposure by the demonstration of sulphur-conjugated pyrrolic metabolites of the alkaloid in circulating haemoglobin.. Aust Vet J 1993;70:312–313.
            doi: 10.1111/j.1751-0813.1993.tb07985.xpubmed: 8216101google scholar: lookup
          19. Lame MW, Morin D, Jones AD, Segall HJ, Wilson DW. Isolation and identification of a pyrrolic glutathione conjugate metabolite of the pyrrolizidine alkaloid monocrotaline.. Toxicol Lett 1990;51:321–329.
            doi: 10.1016/0378-4274(90)90075-Wpubmed: 2111054google scholar: lookup
          20. Estep JE, Lame MW, Jones AD, Segall HJ. N-acetylcysteine-conjugated pyrrole identified in rat urine following administration of two pyrrolizidine alkaloids, monocrotaline and senecionine.. Toxicol Lett 1990;54:61–69.
            doi: 10.1016/0378-4274(90)90056-Rpubmed: 2123045google scholar: lookup
          21. Reed RL, Miranda CL, Kedzierski B, Henderson MC, Buhler DR. Microsomal formation of a pyrrolic alcohol glutathione conjugate of the pyrrolizidine alkaloid senecionine.. Xenobiotica 1992;22:1321–1327.
            pubmed: 1492424
          22. Lame MW, Jones AD, Morin D, Wilson DW, Segall HJ. Association of dehydromonocrotaline with rat red blood cells.. Chem Res Toxicol 1997;10:694–701.
            doi: 10.1021/tx960173vpubmed: 9208177google scholar: lookup
          23. Khan I, Dantsker D, Samuni U, Friedman AJ, Bonaventura C, Manjula B, Acharya SA, Friedman JM. Beta 93 modified hemoglobin: kinetic and conformational consequences.. Biochemistry 2001;40:7581–7592.
            doi: 10.1021/bi010051opubmed: 11412112google scholar: lookup
          24. Kim HY, Stermitz FR, Coulombe RA. Pyrrolizidine alkaloid-induced DNA-protein cross-links.. Carcinogenesis 1995;16:2691–2697.
            doi: 10.1093/carcin/16.11.2691pubmed: 7586188google scholar: lookup
          25. Coulombe RA, Drew GL, Stermitz FR. Pyrrolizidine alkaloids crosslink DNA with actin.. Toxicol Appl Pharmacol 1999;154:198–202.
            doi: 10.1006/taap.1998.8552pubmed: 9925804google scholar: lookup
          26. Lame MW, Jones AD, Wilson DW, Dunston SK, Segall HJ. Protein targets of monocrotaline pyrrole in pulmonary artery endothelial cells.. J Biol Chem 2000;275:29091–29099.
            doi: 10.1074/jbc.M001372200pubmed: 10875930google scholar: lookup
          27. Lame MW, Jones AD, Wilson DW, Segall HJ. Monocrotaline pyrrole targets proteins with and without cysteine residues in the cytosol and membranes of human pulmonary artery endothelial cells.. Proteomics 2005;5:4398–4413.
            doi: 10.1002/pmic.200402022pubmed: 16222722google scholar: lookup
          28. Hansen BT, Jones JA, Mason DE, Liebler DC. SALSA: a pattern recognition algorithm to detect electrophile-adducted peptides by automated evaluation of CID spectra in LC-MS-MS analyses.. Anal Chem 2001;73:1676–1683.
            doi: 10.1021/ac001172hpubmed: 11338579google scholar: lookup
          29. Mattocks AR, Jukes R, Brown J. Simple procedures for preparing putative toxic metabolites of pyrrolizidine alkaloids.. Toxicon 1989;27:561–567.
            doi: 10.1016/0041-0101(89)90117-7pubmed: 2749755google scholar: lookup
          30. Tepe JJ, Williams RM. DNA cross-linking by a phototriggered dehydromonocrotaline progenitor.. J Am Chem Soc 1999;121:2951–2955.
            doi: 10.1021/ja983894kgoogle scholar: lookup

          Citations

          This article has been cited 4 times.
          1. Satué K, Miguel-Pastor L, Chicharro D, Gardón JC. Hepatic Enzyme Profile in Horses. Animals (Basel) 2022 Mar 29;12(7).
            doi: 10.3390/ani12070861pubmed: 35405850google scholar: lookup
          2. González Medina S, Hyde C, Lovera I, Piercy RJ. Detection of equine atypical myopathy-associated hypoglycin A in plant material: Optimisation and validation of a novel LC-MS based method without derivatisation. PLoS One 2018;13(7):e0199521.
            doi: 10.1371/journal.pone.0199521pubmed: 29969503google scholar: lookup
          3. Di Girolamo F, D'Amato A, Lante I, Signore F, Muraca M, Putignani L. Farm animal serum proteomics and impact on human health. Int J Mol Sci 2014 Sep 1;15(9):15396-411.
            doi: 10.3390/ijms150915396pubmed: 25257521google scholar: lookup
          4. Bundgaard L, Jacobsen S, Sørensen MA, Sun Z, Deutsch EW, Moritz RL, Bendixen E. The Equine PeptideAtlas: a resource for developing proteomics-based veterinary research. Proteomics 2014 Mar;14(6):763-73.
            doi: 10.1002/pmic.201300398pubmed: 24436130google scholar: lookup
          Subscribe to our newsletter
          Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.