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Investigation Into the Cause of Australian Stringhalt

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Investigation Into the Cause of Australian Stringhalt Investigation into the Cause of Australian Stringhalt RIRDC Publication No. 11/127 RIRDCInnovation for rural Australia Investigation into the Cause of Australian Stringhalt by Charles El-Hage December 2011 RIRDC Publication No 11/127 RIRDC Project No PRJ-003742 © 2011 Rural Industries Research and Development Corporation. All rights reserved. ISBN 978-1-74254-306-2 ISSN 1440-6845 Investigation into the aetiology of Australian stringhalt Publication No. 11/127 Project No. PRJ-003742 The information contained in this publication is intended for general use to assist public knowledge and discussion and to help improve the development of sustainable regions. You must not rely on any information contained in this publication without taking specialist advice relevant to your particular circumstances. While reasonable care has been taken in preparing this publication to ensure that information is true and correct, the Commonwealth of Australia gives no assurance as to the accuracy of any information in this publication. The Commonwealth of Australia, the Rural Industries Research and Development Corporation (RIRDC), the authors or contributors expressly disclaim, to the maximum extent permitted by law, all responsibility and liability to any person, arising directly or indirectly from any act or omission, or for any consequences of any such act or omission, made in reliance on the contents of this publication, whether or not caused by any negligence on the part of the Commonwealth of Australia, RIRDC, the authors or contributors. The Commonwealth of Australia does not necessarily endorse the views in this publication. This publication is copyright. Apart from any use as permitted under the Copyright Act 1968, all other rights are reserved. However, wide dissemination is encouraged. Requests and inquiries concerning reproduction and rights should be addressed to the RIRDC Publications Manager on phone 02 6271 4165. Researcher Contact Details Charles El-Hage The University of Melbourne 250 Princes Highway Werribee VIC 3030 Phone: 03 9731 2268 Fax: 03 9731 2246 Email: [email protected] In submitting this report, the researcher has agreed to RIRDC publishing this material in its edited form. RIRDC Contact Details Rural Industries Research and Development Corporation Level 2, 15 National Circuit BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6271 4100 Fax: 02 6271 4199 Email: [email protected]. Web: http://www.rirdc.gov.au Electronically published by RIRDC in December 2011 Print-on-demand by Union Offset Printing, Canberra at www.rirdc.gov.au or phone 1300 634 313 ii Foreword Australian Stringhalt (AS) is a debilitating condition of pasture based horses resulting in an abnormal gait and involuntary hyper (over) flexion of the hindlimbs when attempting to walk. It occurs in individual horses or in outbreaks. Prolonged dry summers on poorer quality soil types with unimproved pasture are recognised risk factors. The condition has been documented for over 120 years in south-eastern Australia and has been documented in several continents with little progress in determining the exact cause. There are several common factors that are well reported in outbreaks of AS. They include a strong association with the presence of the flatweed Hypochaeris radicata. Most cases occur in late summer/autumn following prolonged dry spells with affected paddocks being typically of poorer unfertilised soil types. Although a strong association between the disease and H. radicata is well recognised there has been little published work to investigate potential toxins within or associated with the weed. Extensive testing was carried out in this study to identify possible fungal toxins in flatweed associated with outbreaks of AS. No evidence of such mycotoxic alkaloids was found. This finding suggests that such alkaloids are unlikely to be responsible for the nerve damage observed in horses suffering from AS. While further work to identify a specific toxin continues, horse owners could take steps to improve pastures to reduce the prevalence of H. radicata within paddocks where it is identified as a predominant species. Funding for this project was made available from the RIRDC and the University of Melbourne’s Faculty of Veterinary Science, Department of Land and Food Resources and Metabonomics Australia, School of Botany. This report is an addition to RIRDC’s diverse range of over 2000 research publications and it forms part of our Horse R&D program, which aims to assist in developing the Australian horse industry and enhancing its export potential. Most of RIRDC’s publications are available for viewing, free downloading or purchasing online at www.rirdc.gov.au. Purchases can also be made by phoning 1300 634 313. Craig Burns Managing Director Rural Industries Research and Development Corporation iii About the Author Charles El-Hage is a researcher and lecturer in the Department of Veterinary Science at the University of Melbourne. Victoria Australia. He has co-authored several articles relating to intoxication of horses and livestock including: • C El-Hage, MJ Lancaster. (2004). Mycotoxic nervous disease in cattle fed sprouted barley contaminated with Aspergillus clavatus. Australian Veterinary Journal, 82(10): 639-641 • CM El-Hage, LE Bradbury AF Dredge and J.McGregor (2010). A fatal outbreak of suspected botulism in twelve horses. Australian Equine Veterinarian. 29(1) Autumn 2010:50-54 Acknowledgments This project was made possible largely due to the support of Professor Ron Slocombe. The support and assistance of Dr.Daniel Dias, Terra Stark, Nirupama Jayasinghe at Metabolomics Australia (School of Botany), The University of Melbourne. Dr Simone Rochfort DPI Victoria Agribiosciences, La Trobe University, Larry Walker SSPL Hamilton and Angus Tye of the Faculty of Land and Food Resources, University of Melbourne was invaluable. Abbreviations AS Australian Stringhalt BSTFA N,O-bis(trimethylsilyl)trifluoroacetamide) GC Gas Chromatography H. radicata Hypochaeris radicata HPLC High Performance Liquid Chromatography MS Mass Spectrometry NMR Nuclear Magnetic Resonance RH Relative Humidity SPE Solid Phase Extraction TIC Total Ion Chromatogram TMCS Trimethylchlorosilane TMS Trimethylsilyl iv Contents Foreword ............................................................................................................................................... iii About the Author.................................................................................................................................. iv Acknowledgments................................................................................................................................. iv Abbreviations ........................................................................................................................................ iv Executive Summary ............................................................................................................................ vii Introduction ........................................................................................................................................... 1 Objectives ............................................................................................................................................... 3 Methodology .......................................................................................................................................... 4 Sample Collection - Hypochoeris radicata ...................................................................................... 4 Test Batches ............................................................................................................................... 4 Control Batches .......................................................................................................................... 4 Mycotoxin screening ......................................................................................................................... 5 Growth Chamber Experiment .................................................................................................... 5 Comparative GC-MS Analysis of Hypochoeris radicata .................................................................. 6 Sample preparation ..................................................................................................................... 7 Analytical instrumentation ......................................................................................................... 7 Results .................................................................................................................................................... 8 Sample collection .............................................................................................................................. 8 Growth Chamber experiment ............................................................................................................ 8 Mycotoxin analysis ........................................................................................................................... 9 Comparative GC-MS Analysis of Hypochoeris radicata ................................................................ 10 Implications.......................................................................................................................................... 16 Recommendations ............................................................................................................................... 17 References
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