RIRDC Horse Projects completed in 2006-07 and Horse Research in

Progress as at June 2007

November 2007 RIRDC Publication No 07/011

ii

© 2007 Rural Industries Research and Development Corporation. All rights reserved. ISBN 1 74151 411 8 ISSN 1440-6845 RIRDC Horse Projects completed in 2006-07 and Horse Research in Progress as at June 2007

Publication No. 07/011 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.

Dr Nigel Perkins Rural Industries Research and Development Corporation Level 2 15 National Circuit BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 07 4632 0636 Fax: 07 3844 8374 Email: nigel@ausvet.com.au RIRDC Publications Manager Cecile Ferguson Rural Industries Research and Development Corporation Level 2 15 National Circuit BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: (02) 6271 4165 Fax: (02) 6271 4199 Email: cecilef@rirdc.gov.au Website: http://www.rirdc.gov.au Published in November 2007

iii

Foreword This report contains all entries from continuing and completed horse research projects funded by RIRDC in 2006–2007. It also contains completed projects from previous years. The Horse R&D Program aims to assist the development of the Australian horse industry and enhance its export potential. This report is an addition to our extensive catalogue of over 1700 research reports which are available for viewing, downloading or purchasing online through our website: • downloads at www.rirdc.gov.au/fullreports/index.html

• purchases at www.rirdc.gov.au/eshop

Peter O’Brien Managing Director Rural Industries Research and Development Corporation

iv

Contents

2.4 HORSES RESEARCH IN PROGRESS in 2006-2007

PROJECT No

PROJECT TITLE

RESEARCHER

PHONE

ORGANISATION

PAGE No

To improve the management of growing and developing horses UM-70A The role of macrophages in recovery from exercise

induced pulmonary haemorrhage (EIPH) Dr Peter Finnin (03) 9731 2270 The University of Melbourne 1

AVS-1A Short term and future athletic performance of critically ill equine neonates

Dr Jane Axon (02) 6545 1433 Axon Veterinary Services 2

To improve the overall health and welfare of the horse QUT-11A Sacroiliac kinematics in horses Dr Jan Jasiewicz (07) 3864 5821 Queensland University of

Technology 3

UQ-121A DNA typing of urine samples to confirm the donor identity\

Dr Ann Trezise (07) 3365 3647 The University of Queensland 4

UQ-125J Investigation of causal factors for equine amnionitis and foetal loss

Dr Judy Cawdell-Smith (07) 5460 1166 The University of Queensland 5

UQ-132A Pasture management guidelines to minimise risk of laminitis in horses

Dr Christopher Pollitt (07) 3365 2063 The University of Queensland 6

SVH-2J Treatment of placentitis in thoroughbred mares: efficacy of altrenogest

Dr Joan Carrick (02) 6545 1333 Scone Veterinary Hospital 7

UWA-101A Neuromuscular stimulation to reduce joint loading in the equine forelimb

Dr Rochelle Nicholls (08) 9431 3862 University of Western Australia 8

UQ-118A Laminitis preventive and therapeutic strategies

Dr. Christopher Pollitt (07) 3365 2063 The University of Queensland 12

To keep Australia free from major exotic equine disease out-breaks and limit the impact of endemic diseases EQS-1A Placentitis: A major cause of late term foetal loss in

thoroughbred mares Dr Joan Carrick (07) 4691 0164 Equine Specialist Consulting 13

UQ-129A Identification of horses with resistance to small strongyle infections

Dr Glen Coleman (07) 3365 2119 The University of Queensland 14

Respiratory Diseases-Improving the Respiratory Health of Horses US-118A Development of improved treatment and prevention

strategies for inflammatory airway disease of horses

Dr Jennifer Hodgson (02) 4655 0760 Monash University 15

To encourage informed use of modern genetic technology and techniques UMO-36A The development of horse embryonic stem cells

(eESCs) Prof Alan Trounson (03) 9905 0771 Monash University 17

To improve the level of safety in the industry through focusing on occupational health and safety RNS-1A Health and safety in Australian racing Mr Peter V'Landys (02) 9551 7500 Racing NSW

18

UF-11A SHARE - Safety for horses and riders in eventing Dr Raymond Cripps (08) 8201 7627 Flinders University of South Australia

19

UM-58A Epidemiology and risk factor analysis of racetrack fatalities

Prof. Andrew Clarke (03) 9731 2314 The University of Melbourne 20

Information Technology for the Horse Industry UM-66A A prospective study of pre sale radiographs of

thoroughbred yearlings

Prof Andrew Clarke (03) 9731 2314 University of Melbourne 23

Other UQ-115A Role of bacteria and lactic acid in the pathogenesis of

gastric ulceration Dr Rafat Al Jassim (07) 5460 1521 University of Queensland 25

RWE-2A Tracheal stethoscopy: An effective tool for diagnosing DDSP and evaluating treatment?

Mr Robert Curtis (02) 4757 4876 Randwick Equine Centre 26

v

2.4 HORSES COMPLETED PROJECTS in 2006-2007

PROJECT No

PROJECT TITLE

RESEARCHER

PHONE

ORGANISATION

PAGE No

To improve the management of growing and developing horses UMU-33A Risk factor for gastric ulceration in thoroughbred

racehorses A/Prof Guy Lester (08) 9360 7676 Murdoch University 27

To improve the overall health and welfare of the horse RAS-1A An investigation into the detection of three common

veterinary pharmaceuticals Dr John Vine (03) 9376 6760 Racing Analytical Services Ltd 28

To keep Australia free from major exotic equine disease out-breaks and limit the impact of endemic diseases UCS-35A Investigating the role of impaired glucose uptake in

laminitis Dr Martin Sillence (02) 6933 2205 Charles Sturt University 32

UM-71A Integrated pest management for the horse farm

Prof Ary Hoffmann (03) 9479 2769 La Trobe University 34

US-108A Prospective Longitudinal Study of disease on Equine Stud Farms

Dr James Gilkerson 0409 583 169 The University of Melbourne 36

Other UQ-111A Epaxial musculature and its relationship with back

pain in the horse Dr Catherine McGowan (07) 5460 1521 University of Queensland 37

AUR-1A National standards for track monitoring, assessment and design

Mr Arthur Stubbs (03) 9844 1135 Australian Racecourse Managers Association Inc

38

vi

2.4 HORSES

COMPLETED PROJECTS 2005-06 PROJECT No

PROJECT TITLE

RESEARCHER

PHONE

ORGANISATION

PAGE No

Information Technology for the Horse Industry US-124A Communications and information programs for the

horse industry 5 Ms Shirley Ray (02) 9352 6932 University of Sydney 39

To improve the management of growing and developing horses US-129A Risk factors for injuries in thoroughbred racehorses

during commercial training

Prof David Evans (02) 9351 2474 University of Sydney 40

To improve the overall health and welfare of the horse OFF-1A The production of a poster and booklet on

poisonous weeds for horses

Ms Mellisa Offord (02) 8901 0622 42

To more effectively quantify the environmental impact on the horse industry UQ-119A Transfer of weeds to Qld native forests on horses

hooves

Dr. Christopher Pollitt (07) 3365 2063 The University of Queensland 43

Respiratory Diseases-Improving the Respiratory Health of Horses UM-67A Effects of Stud Management on Ecology of Virulent

Rhodococcus equi and Further studies of the ecology of virulent R. equi on horse studs - 2002 stud season

Dr Glenn Browing (03) 8344 742 University of Melbourne 44

Lameness and Limb Injury-Developing More Effective Methods for Prevention, Diagnosis and TreatmentUQ-100A Pasture fructan concentration as a cause of equine

laminitis

Dr. Christopher Pollitt (07) 3365 2063 The University of Queensland 46

Other EPS-1A Prototype Environmental Management System

(EMS) for the horse keeping community - implementation, auditing and monitoring

Ms Jacqueline Frizenschaf (08) 8139 9919 Environment Protection Authority SA

50

1

Research in Progress – To improve the management of growing and developing horses Project Title

The role of macrophages in recovery from exercise induced pulmonary haemorrhage (EIPH)

RIRDC Project No.:

UM-70A

Start Date: 01-Aug-2004 Finish Date: 31-Aug-2007 Researcher: Dr Peter Finnin Organisation: The University of Melbourne Phone: (03) 9731 2270 Fax: (03) 9731 2366 Email: pfinnin@unimelb.edu.au Objectives

• Identify the major mechanisms of chronic lung lesions induced by

EIPH. • Develop methods for reducing chronic ling damage by

modulating macrophage-related events after episodes of EIPH. • Use a range of diagnostic imaging modalities to document

changes to lungs following EIPH, in order to improve in vivo diagnosis of EIPH.

Current Progress

Macrophages play a key role in the control of inflammatory responses. Previous work has demonstrated that the presence of blood within the lung provokes a macrophage dominated inflammatory response, and that this is associated with the development of pulmonary fibrosis.

We have used microarray technology to analyse changes in the level of expression of approximately 100 genes known to be important in immune functions in equine alveolar macrophages. The microarray analysis identified a number of genes that were either up- or down-regulated by equine alveolar macrophages actively involved in in vitro phagocytosis of red blood cells.

Quantitative real time PCR (qRT-PCR) was then used to validate the results of microarray experiments. RNA was extracted from alveolar macrophages actively phagocytosing red blood cells. After conversion of RNA to cDNA, qRT-PCR techniques were employed to follow the level of gene expression over time after exposure to red blood cells, of five genes identified by micorarray as being differentially regulated. Using this technique, we confirmed that these genes are either rapidly up- or down- regulated in alveolar macrophages phagocytosing red blood cells.

Following confirmation of the micorparray results using qRT-PCR, the genes of interest were isolated and attempts were made to produce recombinant 6-Histidine tagged forms of the proteins expressed by the genes of interest. Four of the genes have been expressed, but the expressed proteins are not readily soluble, even when mildly denaturating conditions are used. We have successfully expressed a soluble form of one of the proteins, which was shown by qRT-PCR to be rapidly and highly upregulated.

We are currently mid-way through a clinical trial using this soluble expressed protein in our model of EIPH. Known segments of lung are inoculated with 35ml of autologous blood with or without cytokine. Although preliminary, initial results are promising, suggestive of accelerated blood clearance from cytokine-treated segments. Further animals are currently being studied to confirm that these findings are repeatable.

2

Research in Progress – To improve the management of growing and developing horses Project Title

Short term and future athletic performance of critically ill equine neonates

RIRDC Project No.:

AVS-1A

Start Date: 01-Sep-2006 Finish Date: 31-July-2008 Researcher: Dr Jane Axon Organisation: Axon Veterinary Services Phone: (02) 6545 1433 Fax: (02) 6545 2355 Email: janeaxon@bigpond.com Objectives

• To determine admission parameters and clinical problems that

will affect the survival of the critically-ill neonatal foal. • To determine whether surviving foals reach their sale potential as

yearlings. • To determine whether surviving foals are able to have one race. • To determine clinical parameters in the critically-ill neonatal foal

which affect their future athletic performance.

Current Progress

The data collation and entry for the NICU foals is in the final stages. The data is being entered in Excel spreadsheets. Consultation with Dr Nigel Perkins determined that a complete year of data including race records should be analysed first to ensure the optimal way of data entry for statistical analysis. The complete data for 2004 will be forwarded to Dr Perkins once the racing records have been entered.

3

Research in Progress – To improve the overall health and welfare of the horse Project Title

Sacroiliac kinematics in horses

RIRDC Project No.:

QUT-11A

Start Date: 30-Nov-2006 Finish Date: 30-Nov-2008 Researcher: Dr Jan Jasiewicz Organisation: Queensland University of Technology Phone: (07) 3864 5821 Fax: (07) 3138 3980 Email: j.jasiewicz@qut.edu.au Objectives

• To identify the magnitude and direction of motion available in both

clinically normal and diseased equine sacoiliac joints • Correlate clinical signs, force plate analysis and three dimensional

kinematics in horses affected by sacroiliac disease • To develop more effective intervention for the management of

sacroiliac disease in the horse, based on the kinematic data.

Current Progress

Study 1 aimed to quantify sacroiliac motion in vitro and correlate the applied forces and is complete. By comparing amounts of movement relative to the same applied force in cadavers we have shown that:

• the greatest relative movements in the sacroiliac joint complex are in the transverse and coronal planes

• the most consistent direction of motion of the joint complex during force application is in the sagittal plane

• the most consistent increase in joint motion after ligament transection are in the sagittal plane, suggesting that the sacrotuberous and dorsal sacroiliac ligaments limit sagittally directed forces.

A biomechanical model of the joint complex is currently being developed using data from studies 1 and 2. Study 2 aimed to quantify sacroiliac motion in vivo. This study has been successfully started with a new collaborator (Prof. Rene Van Weeren - University of Utrecht). Treadmill facilities at University of Sydney enabled data collection from horses at different gaits and during manual-physiotherapy manipulation. Data have been collected using skin and bone fixated markers and are being analysed. Study 2 also validated and quantified skin motion artefacts. This will facilitate study of clinical cases of sacroiliac dysfunction beginning in the second half of this year.

4

Research in Progress – To improve the overall health and welfare of the horse Project Title

DNA typing of urine samples to confirm the donor identity

RIRDC Project No.:

UQ-121A

Start Date: 01-Jul-2005 Finish Date: 30-May-2007 Researcher: Dr Ann Trezise Organisation: The University of Queensland Phone: (07) 3365 3647 Fax: (07) 3365 4899 Email: ann.trezise@uq.edu.au Objectives

• Test various methods for DNA typing of horse urine samples. • Establish and optimise a method that is reliable for drug-test positive

samples. • Validate and refine the procedure for routine operation. • Implement as a standard operating procedure in the industry.

Current Progress

We have developed a method that successfully and consistently isolates high quality DNA from horse urine samples. This is an important milestone in this project as it is well known that urine is not a particularly good source of genomic DNA and it is often difficult to obtain high quality DNA consistently. We have trailed various methods of storage of urine samples prior to DNA extraction and found that refrigeration is the best method of storage. DNA extracted from urine stored at 4°C showed the lowest levels of degradation and produced the highest yield of genomic DNA. Urine that was stored frozen at -20°C provided much lower yields of genomic DNA and overall this DNA showed evidence of more extensive degradation. This important progress will be reported at the Equine Science Symposium June 13th & 14th at the Gold Coast. We have begun optimising the generation of DNA profiles from the DNA extracted from urine and are currently determining the optimal DNA concentration to add to the DNA amplification reagents. The next immediate priority is to obtain both drug-test positive and drug-test negative horse urine that has been collected using the standard collection procedures used by the drug testing authorities to determine whether these procedures introduce any contaminants that impact on the generation of DNA profiles from these samples.

5

Research in Progress – To improve the overall health and welfare of the horse Project Title

Investigation of causal factors for equine amnionitis and foetal loss

RIRDC Project No.:

UQ-125J

Start Date: 01-Jul-2006 Finish Date: 01-Aug-2008 Researcher: Dr Judy Cawdell-Smith Organisation: The University of Queensland Phone: (07) 5460 1166 Fax: (07) 5460 1444 Email: j.cawdellsmith@uq.edu.au Objectives

• To determine whether administration of processionary caterpillars

(Ochrogaster lunifer) to pregnant mares is associated with increased risk of foetal loss and abortion.

• To develop a laboratory animal model for use in the screening and study of agents suspected of causing abortion in horses.

• To determine whether exposure of pregnant laboratory animals to Australian native pennyroyal (Mentha satureioides) results in increased risk of foetal loss and abortion.

Current Progress

Administration of 100 grams of whole processionary caterpillars once a day for 5 days to pregnant mares has been shown to cause abortion with gross pathology and bacteriology similar to that found in Equine Amnionitis and Foetal Loss. Pregnant mares have also been exposed to the shed exoskeleton that has been recovered from caterpillar nests. Mares were administered either 1 gram, 2 grams or 5 grams of exoskeleton once a day for 5 days. Two mares dosed with 5 grams aborted within 6 days of the first treatment, one mare dosed with 2 grams aborted on the 6th day and one mare dosed with 1 gram aborted 60 days after treatment. The aborted foetuses had gross pathology and bacteriology similar to that found in EAFL. Currently we have another cohort of mares pregnant for further experiments.

6

Research in Progress – To improve the overall health and welfare of the horse Project Title

Pasture management guidelines to minimise risk of laminitis in horses

RIRDC Project No.:

UQ-132A

Start Date: 01-Jul-2006 Finish Date: 30-Nov-2007 Researcher: Dr Christopher Pollitt Organisation: The University of Queensland Phone: (07) 3365 2063 Fax: (07) 3365 2351 Email: c.pollitt@uq.edu.au Objectives

• Investigate and raise awareness of the problems of excess

carbohydrate in forage as a risk factor in laminitis • Determine levels of NSC in grass that triggers laminitis • Provide management guidelines for pasture to minimise the risk of

these diseases.

Current Progress

Over 100 pages have been written examining current literature on the sugar, starch and fructan levels in Australian pastures. Background information on grass morphology, physiology and the affects of environmental conditions will help horse owners understand when to limit access to grass when NSC are apt to be higher. Many new photographs and illustrations have been prepared. Information on laminitis especially that associated with insulin resistance will complement the revised booklet on laminitis due later this year. Management guidelines for methods to minimise NSC levels in grass have been written and first draft edits are nearly complete. Samples of various dried forages, collected randomly as examples of NSC in Australian forages, were gathered to be sent to a laboratory in the US that is already well experienced in the necessary procedure.

7

Research in Progress – To improve the overall health and welfare of the horse Project Title

Treatment of placentitis in thoroughbred mares: efficacy of altrenogest

RIRDC Project No.:

SVH-2J

Start Date: 17-Nov-2006 Finish Date: 01-May-2009 Researcher: Dr Joan Carrick Organisation: Scone Veterinary Hospital Phone: (02) 6545 1333 Fax: (02) 6545 2903 Email: joan.carrick@sconevet.com.au Objectives

The primary aim of this study is to compare the treatment of late-term pregnant mares with clinical signs of placentitis using two different treatment regimes: • Antibiotic and anti-inflammatory drugs (Trimethoprim-sulphadimidine

and flunixin meglumine) • Antibiotic and anti-inflammatory drug with progesterone

supplementaion (Trimethoprin-sulpha, flunixin meglumine and 44 mg SID Altrenogest).

Current Progress

The earliest that mares will become eligible for enrolment in the project is June 2007. The local veterinarians and the managers of the local brood mare farms have been contacted about participating in the project. Further contact is planned for late May to June 2007.

8

Research in Progress - To improve the overall health and welfare of the horse Project Title:

Neuromuscular stimulation to reduce joint loading in the equine forelimb

RIRDC Project No.:

UWA-101A

Researcher: Dr Rochelle Nicholls Organisation: University of Western Australia Phone: (08) 9431 3862 Fax: (08) 9431 2701 Email: rochelle@cyllene.uwa.edu.au Objectives

• Develop an instrumented elastic sleeve to deliver low-level vibration

to soft tissues of the equine forelimb • Evaluate the effectiveness of the sleeve in reduction of joint loads

during gait • Develop a commercialisation strategy to market the sleeve within the

racing industries.

Current Progress 1. Develop an instrumented elastic sleeve to deliver low-level vibration to soft tissues of the equine forelimb

Two pairs of prototype sleeves have been designed and built by Dr. Nicholls and Mr. Albert Hansma of SurgiCare Australia, between November 2006 and February 2007. The components of the system are shown in Figure 1. The sleeves consist of a Lycra and Neoprene outer which encloses the carpal joint, and is secured with Velcro straps above and below the joint. Six miniature unbalanced electric motors are stitched into the inner lining of the sleeve. The vibration emitted by these motors is controlled from a central power box, with On/Off switches for each motor and a vibration control dial which regulates the vibration from all motors simultaneously. This box also houses the batteries (2xC size) which power the system.

Figure 1: Components of the instrumented sleeve system, including a power and control box, and six miniature electric motors stitched into a Lycra/Neoprene wraparound sleeve.

The sleeve system is shown fitted to a horse in Figure 2. The sleeve fits snugly without slipping, and the control box is attached to a regular lunge roller. Sufficient cabling is provided to allow a full range of motion during gait.a

9

Figure 2: The sleeve system fitted to the nearside carpal joint.

2. Evaluate the effectiveness of the sleeve in reduction of joint

loads during gait Biomechanical testing of the sleeve system was conducted at the School of Veterinary and Biomedical Sciences at Murdoch University in March 2007. Approval was gained from the Animal Ethics Committees at both Murdoch University and The University of Western Australia. Six Standardbred horses were recruited into the study. All horses were members of the Murdoch University teaching herd housed at the Veterinary School. The horses were aged 3-9 years, and all were geldings. The aim of the testing was to compare the carpal joint kinematics and kinetics for trotting horses with and without the sleeve system. Our hypothesis was that a micro level of stimulation would decrease joint loading during trotting.

Figure 3: Equine surgeon Dr. David Murphy attaches markers to the brachium of a test subject prior to biomechanical testing. The horse is secured in a padded crush.

10

Figure 4: Placement of markers for motion capture. Markers are attached to the hoof, pastern, metacarpus and brachium using adhesive non-stretch tape. Leads connecting to EMG electrodes attached to four muscles in the upper limb are also shown.

Methods:

To quantify kinematics and calculate joint loads, three types of data were required: joint and segment motion in the foreleg, ground reaction forces, and muscle activity. These were collected simultaneously during testing. Horses were hand-trotted across a force plate set into the breezeway of the Equine Barn at Murdoch University. Two highspeed cameras (200 Hz; Peak Motus, California) were used to quantify joint speed and 3-D

11

motion, and a Kistler forceplate captured ground reaction forces from the forelimb. Electromyography (EMG) was used to quantify muscle activity in the forelimb. Prior to testing, horses were secured in a padded crush, and their height, weight, rectal temperature, and behavioural measures were recorded. Spherical wooden reflective markers were attached to the hoof, pastern, metacarpus and brachium of the offside foreleg using adhesive tape (Figure 3, Figure 4). The motion of these markers was tracked by the motion capture system. EMG electrodes were also attached to four superficial muscles in the upper limb: • radial carpal extensor m. • common digital extensor m. • extensor carpi ulnaris m. • deep digital flexor m. As the stimulation level emitted to the sleeve system was required to be subsensory (i.e. undetectable), the individual vibration detection threshold for each horse was determined prior to testing. The sleeve was fitted to the offside foreleg and stimulation increased in known increments. For each increment, two observers gave a rating of the horse’s behaviour using a scale of 0-5, where 0 = no reaction and 5 = vigorous whole-body reaction. A “1” score was taken as detection threshold. During testing, the level of stimulation was set at 10 % below the detectable level. During testing, horses were hand-trotted across a forceplate set into the breezeway of the barn. The horse approached the forceplate from approximately 20 metres, achieving a speed of approximately 4 m/s, and trotting for a further 5-10 metres after contacting the plate. For a successful trial, the horse was required to strike the forceplate with the offside forelimb. Two successful trials were required for each of the three test conditions: • without sleeves (baseline); • with sleeves but nil vibration; • with sleeves plus vibration. Tests were conducted in a random order, and adequate rest between trials and conditions provided. A welfare checklist was conducted for each horse at the conclusion of testing to ensure no undue stress or injury was incurred. Results: The final analysis will be completed in July 2007. A customised processing code is currently being written by Dr. Nicholls to analyse the kinematic and kinetic data. This code will calculate carpal joint loading during gait and compare joint forces and angles in the forelimb between the three test conditions. 3. Develop a commercialisation strategy to market the sleeve

within the racing industries. Since the commencement of this project, we have been liaising with Dr. Marcia Taylor at The University of Western Australia’s Office of Industry and Innovation (OII), who are responsible for the commercialisation of university intellectual property. OII also provided funding for this project. In February 2007, Dr. Taylor departed both the OII and UWA. We have not been notified of any replacement for her. This has resulted in a delay to the progress of commercialisation of our invention. Dr. Nicholls is currently discussing the future of this project with OII directors.

12

Research in Progress - To improve the overall health and welfare of the horse Project Title:

Laminitis preventive and therapeutic strategies

RIRDC Project No.:

UQ-118A

Start Date: 01-Oct-2004 Finish Date: 31-Dec-2007 Researcher: Prof Christopher Pollitt Organisation: The University of Queensland Phone: (07) 3365 2063 Fax: (07) 3365 1899 Email: c.pollitt@mailbox.uq.edu.au Objectives

• To understand how laminitis develops at the molecular level to formulate strategies for diagnosis, prevention and treatment.

• To analyse the lesions of acute and chronic laminitis and develop strategies for diagnosis, prevention and treatment.

Current Progress Lamellar tissues studied 7 days after dosing with laminitis inducing

carbohydrate show significant lamellar lengthening and extensive lamellar fragmentation. This explains why lamellar attachments are dysfunctional in chronic cases. Serial retrograde contrast venography of feet developing chronic laminitis shows diagnostic changes as early as 7 day post dosing. By 7 weeks, venograms show filling defects beneath both the proximal and distal hoof wall. Distorted, inward growing proximal hoof wall and dorsal sole appears responsible for the defects and it is predicted that strategic hoof resection will resolve this. An antibody developed to a basement membrane laminin-5 filament fragment fails to immunostain normal hoof lamellae but does stain laminitis affected tissue thus demonstrating that endogenous proteolytic processing of the lamellar interface occurs during laminitis development. Perfusion of lamellar capillaries via intraosseous perfusion of the distal phalanx has been achieved in cadaver limbs. Material injected into the jugular vein has been recovered from hoof lamellae using our newly developed, painless, lamellar microdialysis system. Horses with intraosseous perfusion screws in place show insignificant lameness. Thus, a practical method for single limb lamellar perfusion has been discovered that will be used to trial enzyme inhibitors as laminitis preventives.

13

Research in Progress – To keep Australia free from major exotic equine disease outbreaks and limit the impact of endemic diseases Project Title

Placentitis: A major cause of late term foetal loss in thoroughbred mares

RIRDC Project No.:

EQS-1A

Start Date: 01-Jan-05 Finish Date: 31-May-2008 Researcher: Dr Joan Carrick Organisation: Equine Specialist Consulting Phone: (02) 6545 1333 Email: joan.carrick@sconevet.com.au Objectives

• The primary objective of this study is to document the development

and consequences of placental abnormalities occurring during the second half of gestation in thoroughbred broodmares.

• To correlate placental changes detected in mares by ultrasound, with gross and histopathological evidence of placentitis.

• To determine the incidence of placentitis in normal thoroughbred mares and in 'high risk' mares, ie mares with histories of late term abortion, premature delivery or delivery of a foal with intrauterine growth retardation.

• To monitor the development and progression of placental abnormalities detected by ultrasound in normal and high risk thoroughbred mares.

• To determine whether equine foetal and neonatal weight and health are correlated with placental abnormalities detected by ultrasound.

Current Progress

There were 106 mares enrolled in the study, of which 93 were followed through to foaling. Each mare had a transrectal and transabdomenal ultrasonographic examination every 2 to 4 weeks throughout the pregnancy. The ultrasonographic findings were recorded and summarised in a excel spreadsheet. Mares were treated with antibiotics, altrenogest and anti-inflammatory drugs based on the results of the ultrasound examination. A complete record of foal health and thorough examination of the placenta was conducted on 80 mares from the study. There were 47 high risk mares and 33 normal mares that had complete data sets.

The foaling rate for the high risk mares over the over the 3 previous foaling seasons was 39%- as a result of the examinations and treatment regime, 94% of these mares produced a live foal, 85 % were healthy or had a minor dysfunction that was managed on the farm. Only 9% of the foals from these mares required intensive care.

The foaling rate for the normal mares over the 3 previous foaling seasons was 85% - as a result of the examinations and treatment 94% of these mares produced live foals, 91% of the foals were healthy or had minor dysfunction that could be treated on the farm. Only 3% of these foals required intensive care.

Repeated ultrasonographic examination and treatment of high risk mares significantly improves the foaling rate

The records of the ultrasonographic examination, the placental pathology and the foal health records are currently being examined to determine whether there is a correlation between the ultrasound findings, placental pathology and neonatal foal health.

14

Research in Progress – To keep Australia free from major exotic equine disease outbreaks and limit the impact of endemic diseases Project Title

Identification of horses with resistance to small strongyle infections

RIRDC Project No.:

UQ-129A

Start Date: 01-Jul-2006 Finish Date: 18-May-2008 Researcher: Dr Glen Coleman Organisation: The University of Queensland Phone: (07) 3365 2119 Fax: (07) 3365 1255 Email: g.coleman@uq.edu.au Objectives

• To investigate the feasibility of developing a simple test (or

combination of tests) for the identification of individual horses' resistance to cyathostome infection.

Current Progress

In a prospective trial looking at variations in parasite burdens between animals, we have enrolled nine properties with a total of 59 horses. Monthly faecal egg counts are being conducted on each animal. The trial is incomplete, but it is clear that when faecal egg counts are ranked for each group then individual animals consistently rank highest (or intermediate, or lowest) for faecal egg count among the group. In a separate, abattoir survey we are examining the correlation between FEC and luminal strongyle worm burden. At this early stage, the correlation between the two parameters is weaker than we had anticipated. Whole blood, hair and serum are being collected from all horses in both trials. These samples will be used to analyse variations in MHC haplotype between animals with different worm burdens, or different susceptibility to infection (as identified in the prospective trial). Sera samples will be used in in vitro assays to determine if the serum from horses of different immune status varies in its ability to inhibit cyathostome larval motility.

15

Research in Progress – Respiratory Diseases - Improving the Respiratory Health of Horses Project Title

Development of improved treatment and prevention strategies for inflammatory airway disease of horses

RIRDC Project No.:

US-118A

Researcher: Priscilla Spendlove, Assoc Prof Jennifer Hodgson and Prof David Hodgson

Organisation: The University of Sydney Phone: 02) 4655 0777 Fax: 02) 4655 1212 Email: jennih@camden.usyd.edu.au Objectives

The development of specific recommendations regarding management strategies and pharmaceutical treatment for the control of Lower Airway Inflammation (LAI) in young performance horses. These guidelines will be of value to owners, trainers and veterinarians and will have positive welfare implications.

Current Progress

In recent years it has become increasingly apparent that lower airway inflammation (LAI) is a common disorder of Australian performance horses, more commonly, young Thoroughbreds. Importantly, it has been recognised that this inflammatory process has the potential to impact upon the ability of these horses to achieve peak levels of performance. Subsequently, management strategies and pharmaceutical intervention options for the prevention, control and treatment of the syndrome were investigated.

Research focussed on examining the relevant risk factors for the development of airway inflammation, such as endotoxin concentrations generated by specific feed and bedding choices and stable environment management. In addition, inhalational pharmaceutical preparations were investigated for their efficacy in the treatment of LAI in young, performance horses housed intensively in stables. Furthermore, drug excretion data were derived for each of the pharmaceutical substances administered.

Specific endotoxin levels generated from various feeds and bedding materials within the breathing zone of the stabled horse were determined. Endotoxin has been elucidated to be a major causative factor attributed to the initiation of airway inflammation in the stabled horse. Thereby, ascertaining contributions of endotoxin load made by individual feeds and bedding materials has lead to recommendations relating to feed and bedding selection, handling, storage and management. In the absence of environmental control, short-term benefits may be gained by the use of pharmaceuticals. However, due to the possible adverse effects of systemic drug administration, in particular corticosteroids, inhalational preparations were investigated. A Randomised Controlled Trial (RCT) was conducted at various Thoroughbred and Standardbred training facilities throughout the Sydney metropolitan area to assess the efficacy of inhaled pharmaceutical preparations for the treatment of LAI. Results indicated that steroidal anti-inflammatory therapy (Flixotide® [fluticasone propionate]) was equally efficacious when compared with bronchodilator (Atrovent® [ipratropium bromide]) and steroidal concurrent therapy for the resolution of airway inflammation and tracheal mucus as experienced with LAI. The use of inhaled medications when administered to performance horses

16

must adhere to the Australian Rules of Racing prohibited substance guidelines (AR178.E). Therefore, drug elimination studies were conducted for both ipratropium bromide and fluticasone propionate. These studies determined appropriate recommendations for with-holding periods to ensure no residue detection in voided urine samples. As expected, the excretion profile of inhaled drugs remains to be much more favourable when compared to systemically or orally administered substances. In conclusion, the results of the studies suggest the appropriate selection of feeds and bedding materials may provide a low cost option for reducing the endotoxin burden within the stable environment and hence reduce the incidence and severity of LAI. In addition, the use of target-specific inhaled pharmaceuticals may provide a low-cost, non-invasive method of medicating horses suffering from LAI and increased tracheal mucus.

This research has provided important advances and continued development for the treatment of respiratory disease in performance horses. However, although inhaled medications provide clinical benefit and short-term relief of symptoms such as cough, airway inflammation and increased tracheal mucus, the long-term implications remain to be fully assessed. Therefore, the successful treatment of LAI requires the coupling of both environmental and pharmaceutical intervention and it remains paramount that the inciting stimulus (ie; endotoxin) be removed from the horse’s environment to prevent reversion to the inflammatory status prior to therapy.

Publications P.Spendlove, J.Hodgson, D.Hodgson, N.Malikides. Exposure of Horses to Total and Respirable Particle Endotoxin Concentrations Generated by Specific Feed and Bedding Materials. Veterinary Comparative Respiratory Symposium (VCRS), October 2004, Montreal Canada.

P.Spendlove, N.Basgallop, J.Hodgson, D Hodgson. Detection of Aerosolised Ipratropium Bromide (Atrovent®) in Equine Urine Post-Administration Using Liquid Chromatography Mass Spectrometry (LCMS). Association Veterinaire Equine Francaise (AVEF), October 2006, Versailles France.

P.Spendlove, J.Hodgson, D.Hodgson. Comparative Efficacy of Aerosol Therapies for the Reduction of Pulmonary Neutrophilia Associated with Lower Airway Inflammation (LAI) in Horses. Association Veterinaire Equine Francaise (AVEF), October 2006, Versailles France.

P.Spendlove, J.Hodgson, D.Hodgson. Randomised Controlled Trial (RCT) Evidence of Aerosol Administration Efficacy for the Resolution of Pulmonary Neutrophilia in Young Performance Horses in Sydney, Australia. Veterinary Comparative Respiratory Symposium (VCRS), October 2006, Jena Germany.

17

Research in Progress – To encourage informed use of modern genetic technology and techniques Project Title

The development of horse embryonic stem cells (eESCs)

RIRDC Project No.:

UMO-36A

Start Date: 01-Jul-2005 Finish Date: 01-Jul-2008 Researcher: Prof Alan Trounson Organisation: Monash University Phone: (03) 9905 0771 Fax: (03) 9905 0780 Email: alan.trounson@med.monash.edu.au Objectives

• The project aims to develop pluripotential equine embryonic stem

cells (eESCs) from morulae, blastocysts by separation and culture of the ICM from the horse blastocyst. This method has been used to derive mouse and primate ESCs that are immortal and capable of differentiation into all the cells of the organism. In addition, we propose to derive equine specific eESCs (ES-ESCs) by the insertion of horse somatic cell nuclei and mitochondria into enucleated mouse, cattle or rabbit eggs to isolate eESCs from xenogenic embryos.

Current Progress

2006: Nine putative horse embryonic stem (ES) cells and 19 trophectoderm cell lines were generated; 1 embryonic and 1 trophectoderm (TE) cell line degenerated. Initial characterisation of 6 putative horse ES and 3 TE cell line been completed. Cell lines were frozen and thawed using both traditional slow freezing and vitrification in open-pulled-straws without loss of cell viability. One cell line from interspecies nuclear transfer (cow oocytes as a recipient) was generated; however this was lost after 12 passages. Reconstruction of horse specific embryos using mouse and rabbit oocytes was not successful. 2007: Six horse embryos were collected. Inner cell masses were isolated and cultured in vitro. Three types of cells were isolated from embryos. Two cell types were able to proliferate, however the third cell type failed to grow in vitro. Putative embryonic stem (ES)-like cells were maintained on gamma irradiated mouse foetal fibroblast feeder cells, the putative trophectoderm (TE) cells were maintained in tissue culture flask without feeder cell. Six putative ES- like cell lines were established. Three putative ES-like cells of were frozen for further proliferation and analyses, and the remaining three putatative ES-like cell lines have been grown for 5 passages. Six putative TE cell lines have been maintained. Immunochemistry with embryonic stem cell markers showed that the putative TE cells expressed the pluripotent stem cell marker, Oct4. The results for karyotyping, embryoid body formation and teratoma testing for the putative horse ES-like cells will be performed at passage 10.

18

Research in Progress - To improve the level of safety in the industry through focusing on occupational health and safety Project Title

Health and safety in Australian racing

RIRDC Project No.:

RNS-1A Start Date: 01-Jul-2006 Finish Date: 31-May-2009 Researcher: Mr Peter V'Landys Organisation: Racing NSW Phone: (02) 9551 7500 Fax: (02) 9551 7501 Email: kreece@racingnsw.com.au

Objectives

• The identification of jockey injuries and the risk factors associated with horse and/or rider falls

• Establish a national thoroughbred web based incident database based on the ARID format

• The evaluation of current and alternative rider safety equipment, including vests and helmets.

Current Progress

Injury research Data gathering has been enhanced with the re-launch of the ARID injury reporting forms in Nov/Dec 06. Collection of new data will compliment the previously collated information, which in turn will provide quantified summary of jockey injuries and risk factors which contribute to horse and rider falls. Veterinarians are also collecting data in relation to Horse injury. This again will be loaded into the ARID system for future analysis.

IT development We previously reported that a comprehensive review of the requirements to modify and adapt the ARID database into an existing racing system (IRIS network) was undertaken. We can now report that the preferred supplier has been selected; - Kanbay Pty Ltd, who reduced their original quote providing savings to the Project team funds. Scoping work has commenced with a 3-4 month completion period targeted.

Review of standards and testing of alternate equipment Since the last report, Associate Professor McIntosh and his team at UNSW have been given intent by the NJSRC to proceed to contract. A revised schedule has been agreed to seek to develop a jockey helmet that will meet the specifications of IS EN 14572:2005 High Performance Helmets for Equestrian Activities. This standard exceeds significantly other world standards for equestrian helmets, to such an extent that to the best of our knowledge a helmet has not yet been manufactured to the requisite standard. The focus will be on design issues and elements, and will involve computer simulations, material testing, design activities, and some limited mock up testing of helmets. At the moment, the exchange of contracts is delayed because of clarity required regarding the Intellectual Property Rights of the outcomes of the project. The Legal departments of Racing NSW (on behalf of the NJSRC), UNSW and RIRDC (Jeff Storer) are formatting an agreed set of principle in order to allow commencement.

Review of Jockey health and safety. One of the key findings of the Committee was the need to standardise weight scales nationally; i.e. the amount of weight allotted to each horse to carry is critical in a jockey’s preparation as race day approaches. This will have an important effect on jockey’s health by minimising the need for jockeys to lose weight in short spaces of time. The aim is to have consistency in the weight scale so that, for example, a jockey does not need to ride at 53kg minimum one week and 49kg the next. We can now report that the revised weight scale has been implemented, effective 1 January 2007.

19

Research in Progress – To improve the level of safety in the industry through focusing on occupational health and safety Project Title

SHARE – Safety for horses and riders in eventing

RIRDC Project No.:

UF-11A

Start Date: 01-Jul-2006 Finish Date: 30-Jun-2007 Researcher: Dr Raymond Cripps

Denzil O’Brien Organisation: Flinders University of South Australia Phone: (08) 8201 7627 Fax: (08) 8374 0702 Email: raymond.cripps@flinders.edu.au Objectives

• Continue national data collection and analysis, providing 5-year

surveillance of injuries to riders and horses incurred during the cross-country phase of eventing.

• Develop reporting tools suitable for National Equestrian Federations to fulfil their international reporting obligations to the International Equestrian Federation (FEI).

• Compare Australian, New Zealand and FEI data on falls and injuries.

Current Progress Routinely collected data from all national and international horse trials throughout Australia for 5-year period (2002-2006), on falls of riders and horses and subsequent injuries. We developed a dedicated data collection and analysis program, SHARE, useful for Australian sports administrators and policy makers, and sought to trial its suitability in other national equestrian organisations (India and New Zealand). Collection and analysis of 5 years’ data provided the basis for the model data system SHARE, used to generate reports on various aspects of risk and safety in eventing. While we were unable to secure participation from India, we were provided with New Zealand data for 2005/2006, and used the model to analyse and compare this data as well as international data from the FEI for 2002 – 2006. Use of SHARE for analysis of 5 years’ data has demonstrated its capacity to collect and generate consistent and reliable data on risks in the sport of eventing. SHARE is capable of use in other National Federations for their international reporting obligations to the international body (FEI), and can provide the Australian NF with consistent data which can inform policy and procedures. Cripps RA ,Pagano H 2002. Monitoring falls during eventing - Horse and rider injuries in the cross-country phase of eventing in Australia. Rural Industries Research & Development Corporation Publication No. 02/082, July 2002. Canberra. Cripps RA ,O'Brien D 2004. Monitoring falls during eventing: Rural Industries Research and Development Corporation Publication No. 04/171.

20

Research in Progress – To improve the level of safety in the industry through focusing on occupational health and safety Project Title

Epidemiology and risk factor analysis of racetrack fatalities

RIRDC Project No.:

UM-58A

Researcher: Prof Andrew Clarke Organisation: The University of Melbourne Phone: (03) 9731 2314 Fax: (03) 9731 2377 Email: afclarke@unimelb.edu.au Objectives

• This project will deliver an improved understanding of the causes of and

risk factors associated with racetrack fatalities. This will translate into strategies to address this complex multifactorial problem. This in turn will result in reduced wastage within the industry and have positive welfare implications.

Current Progress

This project was the first in Australia to combine a prospective post mortem case series and large-scale retrospective observational studies to investigate the causes of and risks associated with racehorse fatality in flat and jump starts. The study was encouraged by the Victorian racing industry in response to industry and community concerns regarding high wastage rates of racehorses as well as jockey injuries and fatalities in racing. The purpose of this study was to identify risk factors that could be modified to reduce the risk of fatality in Thoroughbred racing in Victoria. Dr Lisa Boden was the PhD student working with this project. This project and the results generated by Lisa form the basis of Lisa’s thesis which has been successfully defended. Contact details: "Lisa Boden" lboden@staffmail.ed.ac.uk

The following studies have been achieved as a result of this project: A. Post mortem survey of Thoroughbred mortalities on racecourses in Victoria. B. Risk of fatality and causes of death of Thoroughbred horses associated with racing in Victoria, Australia between 1989 and 2004. C. Risk factors for Thoroughbred racehorse fatality in flat races in Victoria between 1989 and 2004. D. Risk factors for Thoroughbred racehorse fatality in jump races in Victoria between 1989 and 2004. E. Estimated attributable fractions for racehorse fatalities in Victoria.

The results of the post mortem study showed that catastrophic limb injury was the major cause of fatality and that sudden death is a more important contributor to racing fatalities than previously reported in Victoria.

The risk of fatality in flat starts was lower in Victoria (0.44 fatalities per 1000 starts) than that reported in North America (1.4-1.7 fatalities per 1000 starts and the UK (0.8-0.9 fatalities per 1000 starts) but the risk of fatality in jump starts was greater in Victoria (8.3 fatalities per 1000 starts) than that reported overseas (3.9 fatalities per 1000 starts) in North America and 5.6 fatalities per 1000 starts in the UK.

The risk of fatality in flat starts appeared to increase over the 15 year study period but it remains unclear whether this was due to a genuine increase in risk (resulting from changes in such factors as climate or racing practices over time) or to increased stringency of reporting of proportions of fatalities.

The risk of fatality in jump starts remained constant over the study period despite jump racing reviews that recommended changes to hurdle and steeple races to improve safety.

21

The risk of fatality on city racecourses was 2.0 times that on country racecourses.

Risk factors associated with increased odds of fatality in flat starts were horse gender (stallions and geldings compared with mares), one or more starts in the 31-60 day period prior to the case or control start, the going of the track (fast or good surfaces), racecourse location (city compared with country tracks), increasing race length and race year .

Risk factors for jump starts included the number of flat and jump starts accumulated in the racing career prior to the study start, one or more starts in the 1-14 days prior to the case or control start, race type (steeplechase compared with hurdle races), track location (city compared with country tracks) and calendar year. Of the risk factors identified as having high population attributable fraction estimates, the most readily modifiable were: track condition in flat races (fast or good tracks), race type in jump races (steeplechase events) and the number of prior flat starts in racing career for horses in jump races.

Of the risk factors identified as having high population attributable fraction estimates, the most readily modifiable were: track condition in flat races (fast or good tracks), race type in jump races (steeplechase events) and the number of prior flat starts in racing career for horses in jump races.

Jump starts by horses with shorter racing careers were more likely to result in fatality. Over the course of a racing career, horses that started in more flat starts prior to the case or control jump start were more likely to die in the next jump start than horses that had accumulated less flat racing experience. Additionally, increased numbers of jump starts prior to the study start were also associated with increased odds of fatality.

The odds of fatality in jump starts increased with each additional flat start accumulated in the racing career. The average number of career flat starts for horses starting in flat races was 17.2 whereas horses starting in jump races had already accumulated an average of 33 career flat starts. If horses were allowed to start in up to 32 flat starts (and no more) before beginning a jump racing career, the risk of fatality in jump starts may decrease by 50% (approximately eight fatalities per year).

This project identified specific risk factors for fatality in flat and jump racing in Victoria. The results of this study should be utilised to direct further research into starts, horses and races at high risk so that appropriate intervention strategies can be implemented to improve the welfare of horses racing in Victoria. Specific recommendations made to the industry as a result of this project include:

• continuation and expansion of post mortem program to include fatalities from country tracks and the collection of blood from cases of sudden death by racetrack veterinarians

• formalised monitoring of injuries and fatalities • standardisation of track rating methods and review of current policy

regarding racing on ‘good’ tracks • further investigation of differences between risk in hurdles and

steeplechase races • restriction of number of flat starts horses can accumulate before

starting a jump racing career.

Publications A very successful international two day workshop on the prevention of racetrack fatalities and injuries was hosted in Melbourne in 2005. This was the first ever international conference addressing this subject. A copy of the CD of the proceedings of the workshop has been produced. The CD includes all of the PowerPoint presentations from the symposium and has proven to be popular for those unable to attend the symposium. Other international meetings are being planned as a result of this inaugural meeting.

22

This project was the basis of a PhD for Dr Lisa Boden. The thesis has been completed and successfully defended. The following publications have arisen from this project to date:

Boden, L., Charles, J., Slocombe, R., Sandy, J., Finnin, P., McCaffrey, J., Clarke, A. Post mortem survey of Thoroughbred mortalities on racecourses in Victoria. In: Proceedings of the AVA Annual Conference, Canberra, May 2004.

Boden, L., Charles, J., Slocombe, R., Sandy, J., Finnin, P., McCaffrey, J., Clarke, A. Post mortem survey of Thoroughbred mortalities on racecourses in Victoria. Australian Equine Veterinarian Vol. 23 2004, p 148. (Abstract from the Bain Fallon Annual Conference of Australian Equine Practitioners, Darwin, July 2004).

Boden, L., Charles, J., Slocombe, R., Sandy, J., Finnin, P., McCaffrey, J., Clarke, A. Post mortem survey of Thoroughbred mortalities on racecourses in Victoria. In: Proceedings of the Annual Conference of the Australian Society of Veterinary Pathologists, Brisbane, October 2004.

Boden, L., Charles, J., Slocombe, R., Sandy, J., Finnin, P., McCaffrey*, J., Clarke, A. Post mortem survey of Thoroughbred mortalities on racecourses in Victoria. Proceedings of the annual conference of the Australian Veterinary Association, Canberra, May 2004

Boden, L., Charles, J., Slocombe, R., Sandy, J., Finnin, P., McCaffrey*, J., Clarke, A. Post mortem survey of Thoroughbred mortalities on racecourses in Victoria. Australian Equine Veterinarian (23), 148.

Boden, L., Charles, J., Slocombe, R., Sandy, J., Finnin, P., McCaffrey, J.,Morton J., Clarke, A.. Sudden death in racing Thoroughbreds in Victoria, Australia. Equine Veterinary Journal (2005) 37 (3) 269.

Boden,LA. J.A. Charles, K.L. Morgan, J. Morton, T.D.H. Parkin, R.F. Slocombe, J.R. Sandy, A. F. Clarke (2005) Risk factors for racehorse fatalities in flat races in Victoria, Australia. Proceedings of Annual Conference of British Equine Veterinary Association

Boden, LA, J.A. Charles, K.L. Morgan, J.M. Morton, T.D.H. Parkin, J.R. Sandy, R.F. Slocombe, A. F. Clarke (2005) Incidence of fatalities and specific causes of death of racehorses on racecourses in Victoria, Australia between 1989 and 2004 Proceedings of Annual Conference of British Equine Veterinary Association

Boden, LA, J.A. Charles, K.L. Morgan, J.M. Morton, T.D.H. Parkin, J.R. Sandy, R.F. Slocombe, A.F. Clarke (2005) Risk factors for Thoroughbred racehorse fatalities in flat races in Victoria, Australia (1989-2004). Proceedings of Australian Equine Association

Boden, LA, Anderson, GA, Charles, JA, Finnin, PJ, Morgan, KL, Morton, JM, Parkin, TDH, Sandy, JR, Slocombe, RF, Clarke, AF. (2005) Incidence of fatalities of Thoroughbred racehorses in Victoria, Australia between 1989 and 2004. Proceedings of Australian Equine Association

Boden, LA, Charles,JA, Slocombe, RF, Sandy JR, Finnin PJ, McCaffrey, JA*, Morton, JM, Clarke, AF. (2005) Post mortem study of Thoroughbred fatalities in Victoria, Australia between 2001 and 2004. Proceedings of the American Association of Equine Practitioners.

Boden LA. (2007 – in press). Determinants of Thoroughbred Racehorse Fatality in Victoria, Australia. RIRDC Project No. UM-58A.

23

Research in Progress – Information Technology for the Horse Industry Project Title

A prospective study of pre sale radiographs of thoroughbred yearlings

RIRDC Project No.:

UM-66A

Researcher: Prof Andrew Clarke Organisation: The University of Melbourne Phone: (03) 9731 2314 Fax: (03) 9731 2377 Email: afclarke@unimelb.edu.au Objectives

• To identify the incidence of radiographic lesions in yearling

thoroughbred horses presented for sale in Australia. • To provide an objective assessment of the impact these findings

have on the racing careers (e.g. number of starts, earning capacity, age of first start) of the horses involved.

Current Progress

Radiograph repositories were introduced to auction sales in Australia in 2003 for pre-purchase assessment of orthopaedic lesions in Thoroughbred yearlings. There were 3,905 yearlings entered into 8 major national yearling sales in this year. Of these 2,847 had radiographs submitted to the radiograph repository. 2,412 of the 2,847 radiograph sets (81,741 individual x-rays) were made available for the present study. The radiographs were reviewed one of four veterinary specialist radiologists. In addition 167 sets were randomly selected and each radiologist read these sets twice. Sale catalogues and results were provided by the sales companies for all eight sales. The Australian Stud Book was used to verify catalogue information and ascertain if yearlings were named for racing or exported overseas. The i-RIS database was used to retrieve the race records of yearlings at two and three years of age.

Melissa Jackson is the PhD student working with this project. This project and the results generated by Melissa will be the basis of her thesis. Contact details: Melissa Jackson, Equine Centre, University of Melbourne, 250 Princes Highway, Werribee Vic 3030. Ph: 03 9731 2316, Fax: 03 9731 2246, Email: m.jackson2@pgrad.unimelb.edu.au

This project has five major components: 1. Comparison between radiographed and non-radiographed

yearlings relating to sale and two-year-old race performance. 2. Prevalence of radiographically detectable orthopaedic lesions in

radiographed yearlings. 3. Association between lesion presence and race performance at

two and three years of age. 4. Race profile at two and three years of age and an economic

analysis of return on money invested in the purchase of the yearling.

5. An assessment of the quality of radiographs and repeatability of assessments of radiograph quality and lesions by specialist radiologists.

This project will be completed in January 2007. Preliminary findings to date include: 1. Overall 3.5% (2871/81741) of films were considered to be non-diagnostic. In approximately half of the radiographic sets examined the aim of a complete assessment was not achieved due to non-diagnostic or missing films. Care in positioning of FlexLM views of the fore fetlocks, LM views of the hind fetlocks and DMPaLO views of the carpus is especially important for maximising radiographic quality.

24

2. As yearling sale price increases, the likelihood of starting a race and earning prize money to cover the cost of training fees increases. The probability of earning more than the purchase price of the yearling decreases with increasing price category.

The components of the study involving the determination of the prevalence of radiographically detected orthopaedic lesions in yearlings, and the analysis of the association of such lesions with future performance are the largest studies of their kind undertaken. This is the first study to document the prevalence of radiographically detectable orthopaedic lesions in Thoroughbred yearlings in Australia and the first Australian study to estimate the effect, if any, of these lesions on race performance.

The career profile and economic analysis component of the project is the largest and most detailed of its kind ever undertaken in Australia. This project is providing information to the industry that will allow both owners and veterinarians to make more informed decisions relating to the significance of radiographic findings in yearlings.

25

Research in Progress - Other Project Title

Role of bacteria and lactic acid in the pathogenesis of gastric ulceration

RIRDC Project No.:

UQ-115A

Start Date: 01-Aug-04 Finish Date: 05-Jul-07 Researcher: Dr Rafat Al Jassim Organisation: The University of Queensland Phone: (07) 5460 1521 Fax: (07) 5460 1444 Email: r.aljassim@uq.edu.au Objectives

• Establish involvement of lactic acid producing bacteria and their

products (VFA & Lactic acid) in the pathogenesis of gastric ulceration in horses.

• Determine the effect of contrasting dietary regimens on the microbial contribution to the build-up of lactic acid and VFAs in the non-glandular portion of the stomach.

Current Progress

Equine Gastric Ulcer Syndrome is a serious health problem in horses. Horses are currently fed high non structural carbohydrate diets and these carbohydrates are fermented by resident stomach bacteria resulting in by-products, mainly as lactic acid. The potent in vitro effect of VFA on reducing mucosal integrity is well established. However, the effect of lactic acid on the mucosa of the non-glandular stomach has not been evaluated experimentally. In addition little is known about the stomach bacteria that ferment carbohydrates and produce lactic acid. In vitro Ussing chamber system was used to evaluate the effect of lactic acid on the integrity of the mucosal tissue and [14C] mannitol was used to measure tissue permeability. Exposure of non-glandular mucosal tissue to high concentration of lactate (40mM) at pH 1.5 resulted in higher conductivity and higher mannitol permeability than tissues exposed at pH 4.0 and 7.0. Main bacterial species belonged to Prevotella (29%), Clostridium (14%), Pseudomonas (13%), Propionibacterium (13%) and Lactobacillus (11%). Other clones clustered with Escherichia coli, Legionella, Voraxella, and Pasteurella. This is the first report on the effect of lactic acid on the integrity of the non-glandular mucosal tissue and on the bacterial diversity of the stomach of the horse.

26

Research in Progress - Other Project Title

Tracheal stethoscopy: An effective tool for diagnosing DDSP and evaluating treatment?

RIRDC Project No.:

RWE-2A

Start Date: 04-Oct-2006 Finish Date: 15-Oct-2007 Researcher: Mr Robert Curtis Organisation: Randwick Equine Centre Phone: (02) 4757 4876 Fax: Email: robac@ieee.org Objectives

• To determine the repeatability of tracheal stethoscopy as a

measurement of respiratory sounds under field conditions. • To determine if the 'throat support device' alters respiratory sounds

in a horse. • Is DDSP reliably detected by tracheal stethoscopy. • Is the 'throat support device' (TSD) an effective treatment for DDSP

under racetrack conditions as determined by tracheal stethoscopy.

Current Progress

Two sets of tracheal Stethoscope apparatus have been designed and constructed. Stethoscope heads have been improved to give a stereo image across the trachea with lower background noise. Four field tests and three treadmill tests have been conducted and the recording technology has been shown to be effective and reliable. Treadmill tests have so far observed gutteral pouch entrapment and pharyngeal collapse. Results analysis software development is underway using a Matlab environment. There is currently a problem with the University of Sydney’s treadmill, and further treadmill video endoscopy is on hold.

27

Completed Projects - To improve the management of growing and developing horses Project Title:

Risk factor for gastric ulceration in thoroughbred racehorses

RIRDC Project No.:

UMU-33A

Researcher: A/Prof Guy Lester Organisation: Murdoch University Phone: (08) 9360 7676 Fax: (08) 9360 2603 Email: G.Lester@murdoch.edu.au

Objectives

• The specific aim of the proposed research is to examine the relationship between management practices and the point prevalence of gastric squamous mucosal ulceration in Thoroughbred racehorses

• To investigate the relationship between ulcer prevalence and defined horse factors including, but not limited to, age, gender, weeks in work, numbers of races this campaign, number of campaigns, and concurrent medication

• To investigate the relationship between ulcer prevalence and training stable.

Background Gastric ulcer disease is highly prevalent in the performance horse

industries. It has been estimated that between 55 and 90% of racing Thoroughbreds have gastric ulcers. The disease has an enormous economic impact, attributable not only to reduced athletic performance, but also in the cost of diagnosis and treatment. In a previous study we reported a wide variability in the prevalence of gastric ulcer disease across a small number of training stables in Western Australia. This population could provide important information as to the risk factors that are important in the development of gastric ulcers.

Research We examined the stomachs of 402 racing Thoroughbreds under the care of 37 different trainers. The trainers were located in metropolitan, semi-rural, and rural regions within the southwest region of Western Australia. The stomach of each horse was graded for ulceration and horses were subsequently categorized into one of two groups; those with no or mild evidence of disease, and those with either moderate or severe gastric ulcers. A comprehensive survey was conducted for each trainer and for each horse.

Outcomes We again documented a significant difference between trainers with respect to ulcer prevalence. There were a number of factors that were significantly associated with ulcer disease. Factors associated with an increased risk of disease were the length of time in training, trainer-reported difficulty in the maintenance of body condition, evidence of stereotypic or abnormal behaviour (specifically crib-biting), and playing a radio in the stables. Factors that were associated with a reduced risk of ulcer disease included exercising at the same location that the horse was stabled and turnout with other horses.

Implications The results of this study indicate that ulcer disease is a widespread problem and that is likely due to a number of factors. Attention to reducing environmental stress would appear to be a beneficial strategy in reducing the prevalence or severity of disease. Animals that demonstrate problems in weight control, almost certainly due to reduced intake in the face of increasing caloric demands, and those that have abnormal (stereotypic) behaviour should be investigated for disease. These horses may benefit from continual medication and/or a change in environment.

Publications Lester G.D., Robertson I. and Secombe C. (in press). Risk Factors for Gastric Ulceration in Thoroughbred Racehorses. RIRDC Project No. UMU-33A.

28

Completed Projects - To improve the overall health and welfare of the horse Project Title:

An investigation into the detection of three common veterinary pharmaceuticals

RIRDC Project No.:

RAS-1A

Researcher: Dr John Vine Organisation: Racing Analytical Services Ltd Phone: (03) 9376 6760 Fax: (03) 9376 6875 Email: jvine@rasl.com.au Objectives

• The purpose of this research is to study the absorption, distribution,

metabolism and excretion of 3 therapeutic substances, frusemide, betamethasone and triamcinolone acetonide, commonly used by veterinarians during the preparation of horses for competition in the thoroughbred and standardbred industries as will as in high level equestrian events.

• Frusemide is a high ceiling diuretic commonly used in the management of exercise induced pulmonary haemorrhage in racehorses. Batemethasone and trimcinolone acetetonide are corticosteroid drugs used for intra-articular treatment and/or management of horses with degenerative joint disease.

• Several of the analytical procedures used to detect these drugs in urine samples from competing horses are abut to change. These changes may alter the detection periods of these commonly-used drugs. The proposed project will provide reliable and up-to-date drug excretion data for frusemide practising veterinarians in a meaningful number of horses. Pilot studies involving the corticosteroid drugs are aimed at providing preliminary data on likely detection periods. This will be a first step in providing information on responsible use of intra-articular corticosteroids which is consistent with animal welfare and the racing industry's rules on the use of medications in racehorses.

Background In recent years the aim of the racing industry regulators to ensure “drug-

free” competition and the need for veterinarians to properly treat racing injuries have increasingly come into conflict. The development of more sensitive analytical methods, necessary to detect today’s sophisticated “go-fast” drugs, may result in the detection of miniscule and pharmacologically irrelevant quantities of genuine therapeutic agents. The detection of such materials inevitably and in some cases unjustifiably reflects poorly on the racing industry and its participants. Reputations can be ruined and severe financial consequences may result from a trace of a relatively innocuous veterinary medication.

Research Robust and properly validated detection methodology has been applied to the measurement of these three drugs in blood and urine. Methods based on gas chromatography mass spectrometry and liquid chromatography mass spectrometry have been developed to ensure that the data generated are as definitive and reliable as possible and that experimental protocols are aligned with those being used to carry out related studies in other countries. This ensures that data will be transferable and enables the range of drugs for which reliable excretion data are available to be greatly extended.

Outcomes It has been shown that after intra-articular administration, betamethasone may be detected for up to three days in urine, albeit at very low concentration. There is no evidence of any significant retention of the drug in synovial fluid and hence it is unlikely that its duration of action is

29

prolonged by this route of administration. The frusemide study has shown that this drug may be reliably detected up to 48 hours in both blood and urine. Data collected on urine void volumes and urine specific gravity suggests that the diuretic effect would not extend beyond that period. Therefore a 48 hour detection time can be reported with a high level of confidence, given that inter-subject variability was relatively small. The study of triamcinolone acetonide administered by intra-articular injection has demonstrated that this drug and its metabolite can be detected for up to 130 and 170 hours, respectively, provided that methods of appropriate sensitivity are applied. The detection times are roughly equivalent to the time for which hydrocortisone is suppressed suggesting that they may also correlate with the duration of action of the drug.

Implications These data can now be used by veterinarians to make more informed decisions about the treatment of horses, particularly in the weeks or days leading up to a race. The data will also assist the racing authorities to draw conclusion about purported treatment regimens in the event that one of these drugs is detected in a pre- or post-race sample.

Publications Vine et al. (2007). Detecting Three Common Drugs in Horses: Frusemide, betamethasone and triamcinolone acetonide. RIRDC Publication No 07/127.

30

Completed Projects - To improve the overall health and welfare of the horse Project Title:

Penetration of pharmacological agents through equine skin

RIRDC Project No.: UQ-114A Researcher: Paul Mills and Sheree Cross Organisation: School of Veterinary Science

University of Queensland Phone: (07) 3365 2964 Fax: (07) 3365 1255 Email: p.mills@uq.edu.au Objectives

• To develop in vitro methodologies to measure drug movement through equine skin

• To measure the regional differences (e.g. thorax vs axilla vs lower leg) in drug penetration

• To measure transdermal penetration of drugs of interest to the equine industry, including non-steroidal anti-inflammatory drugs (NSAIDs), corticosteroids and analgesics (e.g. fentanyl)

• To determine the effects of vehicles (e.g. propylene glycol) on penetration of drugs in the horse

• To determine the effects of skin damage (e.g. rashes) and altered microenvironment (skin cleansers) on transdermal drug movement in the horse.

Background Topical medications are increasingly used in all species, including the

horse. It is important to understand that there are significant species differences between the rate and extent that any drug formulation will move through skin. This means that formulations developed for one species may not have the same amount of drug (more or less) available after topical application. This can lead to either ineffective treatments or toxicity. It is also important to understand transdermal drug penetration when entering horses in competition because many compounds can move through skin and may be detected in blood and urine samples, leading to infringement of the rules of competition. This project was designed to address this lack of basic data concerning transdermal drug penetration in the horse.

Research In vitro techniques to characterise how much and how quickly a range of clinically relevant drugs can pass through horse skin. The same techniques to then investigate regional differences (does drug penetration differ if applied at a different site), the effect of vehicles (what the drug is dissolved in) and the effects of skin damage on transdermal drug penetration. An in vivo model was then trailed to permit investigations relating in vitro results to the living horse.

Outcomes Anti-inflammatory drugs such as hydrocortisone and methylsalicylate penetrate faster and to a greater extent through skin from the leg, which is useful if applying these drugs for anti-inflammatory activity in the legs of horses. Conversely, fentanyl (pain relief) penetrates better through upper body regions which is where fentanyl patches are normally applied.

There are significant effects of the vehicle on the transdermal penetration of testosterone in the horse, with vehicles containing ethanol or propylene glycol (both commonly found in topical creams and gels) associated with greater drug amounts moving through the skin. An important finding was the effect of damage or change to the skin surface created using common clinical approaches to treating skin, including shaving, application of adhesive tape and cleaning skin which all significantly enhanced the penetration of methylsalicylate through skin.

31

Implications Any stakeholder in the equine industry, from trainers, owners,

veterinarians, officials and regulators should be aware that there are significant species differences between how much and how fast a particular formulation will penetrate skin, meaning that a formulation developed and registered for one species will be unlikely to have the same effect and safety margin when applied to another species, including the horse. Also, horse entering into competition should be aware of the potential for the appearance of drugs in blood and urine collected from their horse following topical application which may lead to infringement of the rules of competition. Stakeholders should also be aware that there are regional differences, meaning that a formulation applied to one part of the body may have a different level of effect if applied elsewhere. Different formulations of the same drug can have significant effects on how much of the drug actually penetrates through the skin and is available for whatever purpose it was designed. Most importantly, changes in the integrity of the skin, such as rashes, abrasions, skin disease or the use of agents to clean the skin, can substantially alter the amount and rate of active drug uptake.

Publications Mills P.C. and Cross S.E. (2006) The effects of equine skin preparation on transdermal drug penetration in vitro. Can J Vet Res 70(4):317-20.

Mills P.C. and Cross S.E. (2006) Transdermal Drug Delivery: Basic Principles for the Veterinarian. Vet J 172(2): 218-233.

Mills P.C. and Cross S.E. (2006) Regional differences in transdermal penetration of fentanyl through equine skin. Res Vet Sci. (in press).

Mills P.C. (2006) Vehicle effects on the penetration of testosterone through equine skin. Vet.Res.Comm. (in press).

Mills P.C. and Cross S.E. (2006) Regional differences in the in vitro penetration of hydrocortisone through equine skin. J Vet.Pharm.Ther. 29: 25-30.

Mills P.C. and Cross S.E. (2006) Regional differences in the in vitro penetration of methylsalicylate through equine skin. Vet.J. (doi 10.1016/j.tvjl.2005.08.023).

Mills P.C., Magnusson B.M. and Cross S.E. (2005) Penetration of a topically applied nonsteroidal anti-inflammatory drug into local tissues and synovial fluid of dogs. Am.J.Vet.Res. 66(7): 1128-1132.

Mills P.C. and Cross S.E. (2006). Penetration of Pharmacological Agents through Equine Skin RIRDC Publication No 07/092.

32

Completed Projects - To keep Australia free from major exotic equine disease out-breaks and limit the impact of endemic diseases Project Title:

Investigating the role of impaired glucose uptake in laminitis

RIRDC Project No.:

UCS-35A

Researcher: Prof Martin Sillence Organisation: Charles Sturt University Phone: (02) 6933 2205 Fax: (02) 6933 2812 Email: msillence@csu.edu.au Objectives

• Our aim is to understand the mechanisms that control glucose

uptake in the hoof, then identify the factors associated with endocrine and metabolic abnormalities that lead to impaired glucose uptake, tissue starvation and laminitis.

Background Laminitis is a painful and crippling disease in which the hoof wall becomes detached from underlying tissue may no longer be able to support the weight of the horse. A wide range of factors can trigger laminitis, but horses and ponies that have high concentrations of insulin are most at risk. High insulin concentrations are a sign of glucose intolerance which can be genetic, or induced by diet, lack of exercise, and certain drugs. In this condition (which is similar to human Type 2 diabetes), the function of proteins which normally transport glucose from the circulation to the tissues become obtunded, so that more insulin than normal is required to activate them. Initially we believed that laminitis occurred when the hoof was starved of glucose, causing the connective tissue to disintegrate. However, once we had established that the hoof was not dependent on insulin for glucose uptake (unlike most other tissues), we began to consider other actions of insulin. Instead of regarding poor glucose uptake as the problem and high concentrations of circulating insulin as a sign, we tested the hypothesis that high concentrations of insulin in the blood were actually the cause of laminitis.

Research Our first experiments focussed on the control of glucose uptake, the nature of glucose transporters and role of insulin in isolated hoof tissue. Subsequently, we examined the effect of high insulin concentrations on the whole body, under conditions where glucose concentrations were kept normal.

Outcomes Lamellar tissue was able to take up glucose independent of insulin, and contained no insulin-responsive glucose transporters. However, the infusion of a large dose of insulin over 72 hours caused laminitis to develop in all five ponies tested. This occurred while glucose concentrations were held constant by means of a continuous glucose infusion.

Implications Our findings reinforce the value of using measurements of circulating insulin concentrations to identify horses at risk of laminitis, particularly when exposed to high carbohydrate diets, suffering Equine Cushing’s Disease, or treated with certain anti-inflammatory drugs. Treating glucose intolerance through diet, exercise or medication, should be a primary goal in restoring insulin concentrations to normal and preventing laminitis. New treatments for acute laminitis will result from further studies into the mechanism of action of insulin.

Publications Asplin K.E., Bevan B.E., McGowan C.M., Pollitt C.C. and Sillence M.N.

33

2005. Glucose uptake in the equine hoof. Proceedings of the Nutrition Society of Australia, Dec 2005, Melbourne Vic., Asia Pacific Journal of Clinical Nutrition 14 (Suppl.): S62.

Asplin K.E., McGowan C.M., Pollitt C.C., Curlewis J. and Sillence M.N. 2006.Glucose uptake in the equine hoof. Proceedings of the Australian Equine Science Symposium, June 2006, Gold Coast Qld, Vol. 1, p26.

Asplin K.E., McGowan C.M., Pollitt C.C., Curlewis J. and Sillence M.N. 2007. Role of insulin in glucose uptake in the equine hoof. Proceedings of the American College of Veterinary Internal Medicine 25th Anniversary Forum, June 2007, Seattle WA USA.

Asplin K.E., Sillence M.N., Pollitt C.C. and McGowan C.M. 2007. Induction of laminitis with insulin in healthy ponies. Proceedings of the British Equine Veterinary Association Congress, Sept 2007, Edinburgh UK (In Press).

Sillence M., Asplin K., Pollitt C. and McGowan C. (in press). Investigating the role of impaired glucose uptake in laminitis. RIRDC Publication No 07/158.

34

Completed Projects - To keep Australia free from major exotic equine disease out-breaks and limit the impact of endemic diseases Project Title:

Integrated pest management for the horse farm

RIRDC Project No.:

UM-71A

Researcher: Cindy Edward and Ary Hoffmann Organisation: Centre for Environmental Stress and Research Adaptation

University of Melbourne Vic 3010 Phone: (03) 8344 0017 Fax: (03) 8344 7089 Email: cedward@unimelb.edu.au Objectives

• To complete a research and development project on an alternative to current equine parasite management.

• To develop a biological control program that retains an environmentally sustainable horse farm, by identifying native ground dwelling predators that may break the lifecycle of horse parasites.

Background Horse owners are advised to worm their horses every six to twelve

weeks. The sole reliance on these chemicals to control horse parasites is expensive and detrimental to horses, people and the environment. Resistance in worms to these chemicals is increasing. This leads to higher dosages administered more often.

Research Pest and beneficial invertebrates were monitored in horse paddocks and horse dung with various drenching routines. Exotic dung beetles were released at some of these sites and horse management practices monitored to enable their establishment. Faecal egg counts were used to monitor parasite resistance and burden.

Outcomes A biological control program using an integrated pest management strategy was developed. Horse industry interest was evident through media article feed back. Horse owners proved to be keen to adopt a sustainable strategy in controlling horse parasites and improving productive environmental changes to horse pastures by monitoring parasite burdens, establishing dung beetles and implementing cultural practices and only when necessary using parasiticides. The biological control program will:

• benefit the horse (less chemical administered) • reduce financial costs of worming applications • address chemical resistance • lessen the risk of accidents to the horse handler.

This will maintain a more environmentally friendly and ecologically sustainable horse farm. Faecal egg counts were taken by collecting fresh dung from the trial paddocks. Resistance in worms to ivermectin was found for the first time in horses.

Implications Parasitology in the horse industry should be a priority for further research as new technology and environmental direction is sought. Stud managers, recreational riders and farm workers would benefit from new technology as suggested in recommendations. Workshops, media articles, as well as talks and collaboration with community and Landcare groups are ways to provide the industry with relevant information. The need for inexpensive faecal egg counts and dung beetle commercial supplies is sought by the horse industry. Chemical usage patterns need to be changed to prevent resistance evolution.

35

Publications Edward C.L. and Hoffmann A.A. 2006. Ivermectin resistance in a horse from Australia. Veterinary Record (In press).

Edward C.L. and Hoffmann A.A. 2007. The Effects of Dung Beetle, Bubus bison and Bird Activity on infective larval stages in horse dung of the horse parasite Strongylidae (In preparation).

Media articles: “The Weekly Times”, Leader Newspaper, Veterinary Magazine, LandCare community newsletter, Chaff chat.

Poster: Equitana, CESAR ARC review. Edward C. and Hoffmann A. 2007. Integrated Pest Management for the

Horse Farm RIRDC Publication No 07/090.

36

Completed Projects – Respiratory Diseases - Improving the Respiratory Health of Horses Project Title

Prospective Longitudinal Study of disease on Equine Stud Farms

RIRDC Project No.:

US-108A

Researcher: James Gilkerson Organisation: The University of Melbourne Phone: 03 8344 9969 Fax: 03 8344 7374 Email: jrgilk@unimelb.edu.au Objectives

• To calculate the prevalence, incidence and, where possible, the cost

of a number of diseases, • to produce reports for the participating farms that would allow them to

benchmark the frequency and cost of these diseases on their farms with an industry-based average,

• to allow participating farms to compare the frequency and cost of disease with other participating farms

• to provide the industry with objective data on the frequency and cost of disease upon which to base future research priorities.

Background Prior to 2001 the horse breeding industry in Australia had relied on

anecdote and subjective surveys to determine the frequency of disease and the allocation of research funding into equine diseases. Other rural industries had Government supported surveillance programmes that were funded, in part or in full, by industry funds. The data generated by this project fills a significant gap in our knowledge of the cost and relative significance of disease in the Thoroughbred breeding population.

Research A reporting system was established to collate population data on the resident population of horses on a number of stud farms and to record the diseases for which they were treated. These data were then used to calculate the frequency and where possible the cost of disease to the participating farms. This has enabled some objective measure of the frequency and cost of disease on Thoroughbred farms in Australia to be determined.

Outcomes Estimates of the frequency and cost of disease on participating farms has been determined, as well as the reproductive efficiency and some preliminary investigations into the most common causes of death on participating farms.

Implications This study will allow farm managers, stud veterinarians and equine researchers to make decisions based on an objective estimate of the actual frequency of disease, rather than upon their subjective experience.

Publications Gilkerson J. (in press). Prospective longitudinal study of disease on equine stud farms. RIRDC Project No. US-108A.

37

Completed Projects – Other Project Title:

Epaxial musculature and its relationship with back pain in the horse

RIRDC Project No.: UQ-111A Researcher: Dr Catherine McGowan Organisation: The University of Queensland Phone: 07 54601706 Fax: 07 54601 444 Email: c.mcgowan@uq.edu.au Objectives

• To increase the knowledge of back pain in horses by investigating it using the approach used in humans. Specifically; how pain producing problems in the back are reflected in the deep back muscles and to determine the relationship between decreased size of these muscles and pain and/or poor athletic performance.

Background Back pain and diseases of the spine and pelvis are significant problems

in all types of equine performance causing poor performance, lost training days and wastage. As a result back pain represents a considerable economic and welfare issue for the equine performance industries. Research to date has been led by the veterinary profession with a focus on pathoanatomical problems underlying back pain. However, advances in human back pain have been led by physiotherapy research. Physiotherapy research has focussed on the neuromotor control model and the associated dysfunction especially of the epaxial or deep back muscles that occurs as a result of back pain from different forms of pathology.

Research The anatomy, biomechanics and function of the equine epaxial muscles were described in initial studies using dissection and MRI. Following this, the equine epaxial muscles were measured and analysed for reliability and repeatability using ultrasonography in different groups of normal horses. In the final phase, horses with back pain were examined using ultrasonography and compared to bony pathology found at post mortem.

Outcomes The results of this project have shown that the anatomy and function of the equine epaxial muscles are comparable to that of humans. Ultrasonography was found to be a repeatable and reliable tool for measurement of the equine epaxial muscles and when examined in clinical cases of equine back pathology, there was a clear reduction of the epaxial muscle size at the level of and close to areas of significant injury or pathology.

Implications Ultrasonography of the epaxial muscles is a valuable non-invasive tool that will help detect back pain and associated pathology in horses. This information will be valuable to veterinarians and physiotherapists managing back pain and poor performance syndromes in athletic horses and will lead onto future work on the effect of physiotherapeutic intervention on the recovery of epaxial muscle function following back pain.

Publications Stubbs, N.C., Hodges, P.W., Jeffcott, L.B., Cowin, G., Hodgson D.R. and McGowan C.M. Functional anatomy of the caudal thoracolumbar and lumbosacral spine in the horse. Equine Vet. J. Suppl. 36: 393-399. 2006.

McGowan, C.M., Stubbs, N.C. and Jull G. Review: Equine physiotherapy: a comparative view of the science underlying the profession. Equine Vet J. 2007 Jan 39(1):90-4.

McGowan C., Stubbs N., Hodges P. Jeffcott L. (in press). Back Pain in Horses: Epaxial Musculature RIRDC Publication No 07/118.

38

Completed Projects – Other Project Title:

National standards for track monitoring, assessment and design

RIRDC Project No.: AUR-1A Researcher: Mr Arthur Stubbs Organisation: Australian Racecourse Managers Association Inc Phone: (03) 9844 1135 Fax: (03) 9844 4554 Email: akstubbs@optusnet.com.au Objectives

• Development of national standards for monitoring, assessment and design of Australian race and training tracks, to enable objective investigation of relationships for track design, surfaces and profiles with track performance and user safety.

Background Racecourse Managers Conferences since the mid 1990’s have raised

the issue of a lack of standards for track design and construction in particular, leading to a number of failures of tracks and wastage of considerable expenditure. This led to several studies to explore what standards and objective knowledge existed in this area. It became clear that there was little good data available on which to base sound design. This project was aimed at bringing industry participants together to pool information and encourage an industry wide approach to identification and resolution of standards for national guidance and application.

Research Two workshops were held to firstly summarise current knowledge and identify priorities for standards and then, through investigations by working groups, to more accurately define the standards available and indicate what future work and funding was needed to complete a portfolio of national standards for industry use.

Outcomes Recommendations have been tabled regarding adoption of certain national standards by the industry, and for the direction of future research, funding and monitoring with the ultimate aim of producing a National Master Plan for Track Infrastructure.

Implications The impetus forged by the two workshops and the working groups formed will continue to harness a nation wide collaborative approach to this fundamental issue which is vital to future industry viability and progress.

Publications Australian Racecourse Managers Association Inc (2007) “Australian Racecourses National Track Standards - Monitoring, assessment and design”. RIRDC Publication No 07/159.

39

Completed Projects 2005-2006 – Information Technology for the Horse Industry Project Title

Communications and information programs for the horse industry

RIRDC Project No.:

US-124A

Researcher: Ms Shirley Ray Organisation: The University of Sydney Phone: (02) 9463 9305 Fax: (02) 9463 9100 Email: shirleyr@camden.usyd.edu.au Objectives

To continue to ensure widespread industry support for the RIRDC Equine R&D Program and to enable the results of the R&D to be used by the industry, a strong communication program is necessary to ensure results get to end users. • A database of equine organisations. • A database for communication with the industry. • RIRDC Equine Research News. This newsletter of 8 pages,

published quarterly, will provide the industry with information on topical research from around the world as well as updates on current RIRDC funded projects, interviews with research workers, workshops and invited reviews.

• RIRDC Internet Web Site. An Internet web site for the RIRDC Horse Program which will include links for online purchase of RIRDC publications, download of the RIRDC Equine Research News, search facility for past issues and a subscription facility to receive the RIRDC Equine Research News via the Internet.

Research Each newsletter published had a print run of approximately 9,500 copies, all of which were distributed to the various sectors of the horse industry. The newsletter contained summaries of articles of local and international interest, interviews with researchers in particular fields, announcements of new RIRDC funded projects and updates of all currently funded projects, and items of general interest to the horse industry including RIRDC events and new publications. Articles from the newsletter have been reprinted in many local and international publications including: The Horse (USA), Sport Horse Journal, Breeding and Racing, Race Calendars and Hoofbeats.

The RIRDC Equine Research Program Web site contains articles that have been printed in the RIRDC Equine Research News; these articles are accessible through an index of subject areas. A local search engine allows the user to search the site for a keyword. The site has links to relevant pages of the RIRDC web site, including the Five Year Plan, Objectives and Strategies of the Equine Research Program and RIRDC’s e-shop. A links page includes links to local and international research institutions and other sites of interests. At least 50 websites contain a link to the RIRDC Equine Program website and these include: Cyberhorse, Australian Racing Board; Australian Horse Resources and Australian Horse Net Service. All past issues of the newsletter are available for download in PDF format which can be viewed using Adobe Acrobat. This means that anyone with Internet access can view the newsletter in its colour print format.

The mailing list for the RIRDC Equine Research Program is updated regularly and contains contact details of Australian Horse Associations, Research Institutions and Personnel, Horse Publications and other media and individuals with an interest in the horse industry. The mailing list contains over 2,000 entries

Publications Ray S. and Offord M (2007). Communication and information programs for the horse industry RIRDC Publication No 07/117.

40

Completed Projects 2005-2006 - To improve the management of growing and developing horses Project Title

Risk factors for injuries in thoroughbred racehorses during commercial training

RIRDC Project No.:

US-129A

Researcher: Associate Professor David Evans Organisation: The University of Sydney Phone: (02) 9351 2474 Fax: (02) 9351 3957 Email: davide@vetsci.usyd.edu.au Objectives

• The aim of this study was to investigate the epidemiology of MS

injuries in Australian two- and three-year-old Thoroughbred racehorses. More specifically the aims were to describe the incidence and impact of MS and determine risk factors for MS injuries, in particular shin soreness.

Background Musculoskeletal (MS) injuries have been identified as a common health

problem in Thoroughbred racehorses. The most common MS injuries in two- and three-year olds have been identified as shin soreness and problems involving the carpus and fetlock joints. These MS injuries incur both direct and indirect costs. After reviewing the literature it is clear that there is very limited understanding of the risk factors for MS injuries. In particular, there is almost a complete absence of any investigations that have collected data on daily training in an effort to understand the training related risk factors for MS injuries. Studies that utilize daily training records are fundamental to the understanding of training related risk factors for MS injuries. An understanding of the risk factors for MS injuries would facilitate the design of strategies that may reduce the occurrence and impact of MS injuries.

Research During the study period 451 horses were enrolled during their two-year-old racing season. Training and injury information for these horses were collected directly from the trainer. The research confirmed previous research that MS injuries are a common problem in two- and three-year-old Thoroughbred racehorses. The results showed that injury prior to the first start had considerable impact on time to first start. However, preparations after the first MS injury do not appear to be adversely affected. The investigation of risk factors showed that increasing distance galloped at high speed increased the risk of MS injury.

Outcomes Since 2000 the results of this study have been presented to a number of difference associations and audiences. These were:

• Australian Equine Veterinary Association • Society of Veterinary Epidemiology and Preventive Medicine • International Society Veterinary Epidemiology and Economics • Institute of Veterinary and Biomedical Science, Massey University • Australian and New Zealand racetrack managers • Sydney metropolitan and provincial Thoroughbred trainers • Australian Veterinary Association • British Equine Veterinary Association • International Horse Industry Symposium

Implications The results of this study show that exposure to high-speed exercise is associated with an increased risk of injury. Unfortunately, racehorses must be exposed to high-speed exercise in order to prepare them for competition. Therefore, future research should focus on developing ways in which the risk associated with high-speed exercise can be reduced.

41

Publications Cogger N., Perkins H., Hodgson D.R., Reid S.W.J. and Evans, D.L.

(in press). “Risk factors for musculoskeletal injuries in two-year-old Thoroughbred racehorses.” Preventive Veterinary Medicine. Cogger N., Evans D., Perkins N., Hodgson D and Reid S. (2006). Risk Factors for Injuries in Thoroughbred Racehorses. RIRDC Publication No 06/050.

42

Completed Projects 2005-2006 - To improve the overall health and welfare of the horse Project Title

The production of a poster and booklet on poisonous weeds for horses

RIRDC Project No.:

OFF-1A

Researcher: Ms Mellisa Offord Organisation: Mellisa Offord Phone: (02) 8901 0622 Fax: (02) 9451 3286 Email: mofford@optusnet.com.au Objectives

• To produce a poster and booklet showing plants and trees that are

poisonous to horses.

Outcomes A book and poster have been produced and these were launched at the Equine Science Symposium held on the Gold Coast in June 2006. The book is titled Plants Poisonous to Horses – An Australian Field Guide. It is 127 pages and it is a guide for horse owners to help them prevent plant poisoning in their horses. It clearly outlines the factors that influence the risk of plant poisoning and the ways to reduce this risk. The types of toxins that can be found in plants are discussed, along with the signs of poisoning that may be seen in horses that have ingested toxic plant material. A description of each plant is provided, including structure, growth habit and identifying features. The book contains over 200 colour photos of poisonous plants. Management notes suggest ways of dealing with poisonous plants on properties. The book includes a comprehensive list of ornamental plants that are known to be toxic to horses. The reference list is a useful resource for further reading and includes the journal articles, industry papers, books, CD ROMs and Internet resources that were used to create this publication. The poster highlights 11 plants in Australia that are a risk to horses.

Publications Offord M (2006). Plants Poisonous to Horses: An Australian Field Guide. RIRDC Publication No. 06/048

43

Completed Projects 2005-2006 – To more effectively quantify the environmental impact on the horse industry Project Title

Transfer of weeds to Qld native forests on horses hooves

RIRDC Project No.:

UQ-119A

Researcher: Dr Christopher Pollitt Organisation: The University of Queensland Phone: (07) 3365 2063 Fax: (07) 3365 1899 Email: c.pollitt@uq.edu.au Objectives

• Do horse faeces contain viable seeds which may germinate under

suitable conditions or persist in surface soil. • Are the plants that germinate from horse manure found beside the

trails used by horse riders in forests of SE Queensland?

Background Weeds accidentally introduced since European settlement constitute a threat to the ecological sustainability of native plant communities. Weeds may be introduced into native ecosystems via vehicular, human and animal vectors and there is anecdotal evidence that horse manure contains viable weed seed. A rigorous scientific protocol is necessary to discover if seeds in horse manure are viable and pose a threat when horses traverse areas of conservation significance.

Research Horse faeces were collected from horses grazing an 80 hectare pasture without recent weed control and from race horses maintained in stables. Half of the faeces were dried and half kept wet before being mixed with a commercial, seed free, potting mix.

Outcomes Ten plant species germinated in the pasture derived faeces (compared to one from stabled horses) and there was a higher seed load in the pasture faeces. Dominating seed germination was the grass Queensland Blue Couch (Digitaria ciliaris). Germination was 5 times greater in the dried faeces suggesting that prior drying favours grass seed germination. Only one native species, emu foot (Psoralea tenax) was found. All species that germinated in manure have current weed status.

Implications The faeces of horses grazing weed infested pastures contain viable seed that, potentially, may infest native forests. However further work is required to determine if this actually occurs. To determine the role of horse faeces as a source of weeds, plants growing on 100 m transects of the sampled pasture and transects of the trails in nearby native forests will be compared. When complete these studies will supply objective data to assess the level of threat posed by horse riding in areas of native vegetation and will help formulate rational management strategies for recreational horse riding.

Publications In press

44

Completed Projects 2005-2006 – Respiratory Diseases – Improving the Respiratory Health of Horses Project Title

Effects of Stud Management on Ecology of Virulent Rhodococcus equi and Further studies of the ecology of virulent R. equi on horse studs - 2002 stud season

RIRDC Project No.:

UM-47A and UM-67A

Researcher: Glenn F Browning, James R Gilkerson and Gary Muscatello Organisation: School of Veterinary Science

The University of Melbourne Phone: 03 8344 7342 Fax: 03 8344 7374 Email: glenfb@unimelb.edu.au Objectives

• To develop methods to improve understanding of the factors that make some areas on horse farms a greater risk for development of rattles in foals and apply these to identifying factors that might be addressed by changes in management of farms.

Background Although rattles is an important disease problem in young foals there are

limited options for control. Current methods rely on reducing faecal contamination and dust on studs, but we have little understanding of whether these are the only factors that account for higher infection and death rates on some studs.

Research This project aimed to develop novel methods for assessing the risk associated with particular horse farm environments. These methods were based on bacteriological sampling of air and soil on studs, then identifying those bacteria capable of causing rattles. Once these methods were developed they were applied to examination of different horse farms and different places on those farms. The method that was developed was also used to examine the breath of foals with rattles. This suggested that foals may be able to transmit virulent Rhodococcus equi to each other.

Outcomes The most dangerous areas on studs for foals are likely to be laneways and holding pens. Control may be aided by reducing the time that foals spend in these areas. In addition areas that had sandy, dry and/or acidic soils seemed to be a greater risk. Some control may also be achieved by avoiding close contact between foals, particularly if they are known to be suffering from rattles. Sampling air from foals may be a sensitive diagnostic technique for the early detection of infected foals.

Implications Application of methods developed in this project have provided some suggestions for reducing the incidence of rattles and further use of these methods may enable the benefit obtained from different approaches to control to be assessed. Different approaches to treatment may also be able to be assessed.

Publications Muscatello, G. Rhodococcus equi: the organism and epidemiology. Australian Equine Veterinarian 22:190-197 (2003).

Muscatello, G., and Browning, G. F. - Identification and differentiation of avirulent and virulent Rhodococcus equi using selective media and colony blotting DNA hybridization to determine their concentrations in the environment. Veterinary Microbiology, 100:121-127 (2004). Muscatello, G., Gerbaud, S., Kennedy, C., Gilkerson, J. R., Buckley, T., Klay, M., Leadon, D. P., and Browning, G. F. – Comparisons between

45

concentrations of Rhodococcus equi and virulent R. equi in the air of stables and paddocks on horse breeding farms in a temperate (European) climate. Equine Veterinary Journal, 38:263-265 (2006).

Phumoonna, T., Muscatello, G., Chicken, C., Gilkerson, J. R., Browning, G. F., Barton, M. D., And Heuzenroeder, M. W. - Clinical evaluation of a peptide-ELISA based upon N-terminal B cell epitope of the Vap A protein for early diagnosis of Rhodococcus equi pneumonia in foals. Journal of Veterinary Medicine B, 53:126-32 (2006).

Muscatello, G., Gilkerson, J. R., and Browning, G. F. - Comparison of two selective media for the recovery, isolation, enumeration and differentiation of Rhodococcus equi. Submitted. Muscatello, G., Anderson, G. A., Gilkerson, J. R., and Browning, G. F. - Associations between the ecology of virulent Rhodococcus equi and the epidemiology of R. equi pneumonia on Australian Thoroughbred farms, Applied and Environmental Microbiology, in press (2006).

Muscatello, G., Gilkerson, J. R., And Browning, G. F. - Detection of virulent Rhodococcus Equi in exhaled air samples from foals. Submitted.

Muscatello, G., Gilkerson, J. R., and Browning, G. F. (2006). Rattles in Horses By Gary Muscatello, James R Gilkerson and Glen F Browning. RIRDC Publication No 06-097.

46

Completed Projects 2005-2006 – Lameness and limb injury – Developing more effective methods of prevention, diagnosis and treatment Project Title

Pasture fructan concentration as a cause of equine laminitis

RIRDC Project No.:

UQ-100A

Researcher: Dr Christopher Pollitt Organisation: The University of Queensland Phone: (07) 3365 2063 Fax: (07) 3365 1899 Email: c.pollitt@uq.edu.au Objectives

• The cause of grass founder will be understood and a strategy for diagnosis, prevention and treatment will be available

• A strategy for pasture and grazing management will be available so that horses and ponies do not consume sufficient fructan to develop laminitis.

Research 1. Establish that dosing horses with Raftilose causes laminitis

Horses dosed with Raftilose or oligofructose (OF) at 3 dose rates (7.5;10 & 12.5g/kg) developed clinical and histological laminitis in at least one foot. Higher doses of OF were associated with significantly more severe (grade 3) histological laminitis (p<0.05). All the horses developed profuse diarrhoea at around 18h that ceased by 36-44h. Pyrexia and elevated heart rate peaked between 16 and 20h and returned to near normal by 48h. Faecal pH began to fall within 4 hours of dosing, reaching its lowest value (3.7) at around 18h. Blood D-lactate, of bacterial origin, peaked at 24 hours and then returned to the normal zero. Since all horses developed laminitis, the results establish that a storage carbohydrate, other than starch, can induce the disease. OF is a principal non-structural carbohydrate of pasture grasses, suggesting it is grass OF that causes laminitis and not other substance(s), such as dietary amines. Compared with traditional alimentary overload with starch, OF dosing resulted in reduced morbidity and no deaths, indicating that alimentary overload with fructan is an efficient and more humane experimental model for the induction of acute laminitis.

A paper entitled “Equine laminitis: experimental induction with oligofructose by C.C. Pollitt* and A. W. van Eps was submitted to the Equine Veterinary Journal in mid 2003. The referees were not satisfied that 3 pairs of horses dosed with oligofructose (OF) at 3 dose rates could indicate significant trends (despite our professional statistician’s explanation of the methods used). Publication has been deferred until 4 more horses have been dosed with the median dose (10/kg Bwt OF).

2. Establish surgical caecal cannulation

The difficulties that were encountered with caecal cannulation in 2002 have been overcome by using an alternative 2-step method. In stage 1 an adhesion was created between the caecum and the skin after the muscle in between was surgically removed. In stage 2, 14 days later, access to the caecal lumen was established by trocarisation of the adhesion and a soft, rubber caecal cannula was introduced. One horse has been cannulated (HGF1) and laminitis was induced successfully giving us insight into events in the caecum for the first time. Two horses have had stage 1 surgery and are ready for cannula insertion (stage 2). More cannulations and laminitis inductions are planned for 2005.

3. Establish profile of normal caecal microbiota by culture

47

Profiles of normal caecal microflora populations were documented through the creation of novel habitat-simulating medium containing Raftilose as the sole energy substrate (incubated equine caecal-fluid agar - IECA). Using this media, raftilose utilising organism (RUO) numbers in relation to other anaerobic bacteria were established for normal caecal contents.

4. Purify LTFs by filtration

The trigger factors responsible for initiating laminitis remain elusive. It is clear however that the factors responsible for triggering the onset of laminitis occur in the equine hindgut 8-16h post fructan administration. Furthermore, the dominance of S. bovis, during this period implicates it as the aetiological agent of equine laminitis. Attempts to purify laminitis trigger factors (LTFs) from large volumes of S bovis culture media, using centrifugal filtration equipment, were not successful. We have developed a new casein containing medium and confirmed by zymography that S bovis produces a protease in the presence of casein. Analysis by zymography in gelatin containing PAGE gels showed the S bovis protease (SBP) was a gelatinase and possibly a metalloproteinase. Since the activated MMPs that cause laminitis are also gelatinases we investigated the possibility that SBP was a LTF. SBP was not inhibited by batimastat thus diminishing its likely resemblance to lamellar MMP. Interestingly caecal liquor taken when S bovis proliferation during OF induced laminitis had peaked contained very high concentrations of SBP. SBP and clarified caecal liquor from horses developing laminitis did not induce lamellar separation in vitro suggesting that LTFs are not of bacterial origin. It seems more likely that S bovis proliferation and concomitant protease production damages the mucosal epithelium of the hindgut and that LTFs are generated from the epithelium itself, during the re-epithelialisation process.

5. Design DNA probes to estimate normal caecal ecosystem Fluorescent in situ hybridisation (FisH) probes, derived by 16s rRNA gene sequencing, successfully identified Gram positive and negative organisms in caecal and faecal material from normal horses. The results both validated FisH as a useful tool for studying hindgut bacterial populations and showed potential as a means of analysiing the caecal microflora during laminitis induction.

6. Search literature for fructan data in Australian pasture

A collection of the relevant literature applying to grazing horses has been established. Contact has been made with an expert in Colorado USA (Katy Watts) who is an agronomist specialising in soluble carbohydrates as a cause of laminitis. It would be profitable if Dr Watts was employed by RIRDC to visit Australia for 8 weeks to investigate fructan in Australian grasses and produce a report suitable for Australian horse owners.

Milestone 6 can be achieved employing a specialist agronomist.

7 Extract and cut DNA from microbes associated with laminitis

The implanted caecal fistula enabled analysis of the caecal fluid during the development of laminitis. Rapid proliferation of OF-utilising organisms was observed in all samples prior to the onset of laminitis. Caecal and faecal isolates from horses developing laminitis were subjected to RFLP analysis and formed 10 operational taxonomic units (OTU). Representative isolates from each OTU were sequenced and Streptococcus bovis was demonstrated to be the predominant, culturable, RUO found in the equine hindgut prior to the onset of equine laminitis.

8. Establish profile of laminitis caecal microbiota by culture

Changes in caecal microflora populations were documented using the novel habitat-simulating medium containing Raftilose (IECA). Using this media, changes in raftilose utilising organism (RUO) numbers in relation to other anaerobic bacteria were monitored throughout the course of experimentally

48

induced laminitis. IECA and basal medium 10 (BM10) was used to determine viable counts for both Raftilose utilising organisms (RUO) and total anaerobes every 8 hours during Raftilose induced laminitis.

One hundred and fifty-six RUO were collected throughout the course of fructan-induced laminitis from 5 horses. Rapid proliferation of Raftilose-utilising organisms was observed in all samples prior to the onset of laminitis. Post-laminitis, however, S. bovis was not as prevalent, with Escherichia coli becoming established as the predominant RUO. RUO increased by 2.66 and 5.01 log units in caecal fluid and faeces respectively, while the total anaerobe counts obtained on basal medium 10 (BM10) increased by 1.05 and 1.92 log units in the caecum and faeces respectively. The pH of caecal fluid and faeces decreased to a pH of 4.4 at 8 hours and 4.2 at 16.25 hours and returned to normal within 16 hours in both samples.

9. DNA probes to identify laminitis causing caecal ecosystem

A fluorescence in situ hybridisation (FISH) probe targeted to S. bovis (Sb127) and a few other closely related organisms was developed and applied to the faecal samples collected throughout the course of equine laminitis. Sb127-targeted organisms were demonstrated to increase significantly between 8-16h post-fructan administration in all horses. These organisms then rapidly declined and were no longer observable in faecal specimens by 32-48h. These results both validated FISH as a useful tool for studying changes in known bacterial populations throughout the course of equine laminitis and added further strength to the hypothesis that RUO proliferate rapidly between 8-16h. These studies confirm the rapid proliferation of Streptococcus spp. observed in gram stains and by viable counting.

10. PCR the RNA from microbes associated with laminitis

The PCR products from laminitis associated isolates were subjected to restriction fragment length polymorphism (RFLP) analysis and grouped into operational taxonomic units (OTUs). Isolates with identical RFLP banding patterns were grouped into the same OTU.

11. Quantify fructan fluxes in pasture on daily and annual basis

Since commencing this project data has become available in publications by authors specializing in agronomy and animal production. The recruitment of agronomist Katy Watts (as suggested above) could achieve this milestone.

Publications Pollitt, C.C., and van Eps A.W. (2004). Prolonged, continuous distal limb cryotherapy in the horse. Equine vet. J. 36: 216-220.

Kyaw-Tanner, M. and Pollitt, C. C. Equine laminitis: increased transcription of matrix metalloproteinase-2 (MMP-2) occurs during the developmental phase. Equine vet. J. 36:221-225.

French, K.R. and Pollitt, C.C. (2004). Equine laminitis: loss of hemidesmosomes in hoof secondary epidermal lamellae correlates to dose in an oligofructose induction model: an ultrastructural study. Equine vet. J. 36: 230-235.

French, K.R. and Pollitt, C.C. (2004). Equine laminitis: cleavage of L5 associated with basement membrane dysadhesion. Equine vet. J. 36: 242-247.

van Eps, A.W. and Pollitt, C.C. (2004). Equine laminitis: cryotherapy reduces the severity of the acute lesion. Equine vet. J. 36: 255-260.

French, K.R. and Pollitt, C.C. (2004). Equine laminitis: glucose deprivation and MMP activation induce dermo-epidermal separation in vitro. Equine vet. J. 36: 261-266.

French, K.R. and Pollitt, C.C. (2004) Equine laminitis: congenital,

hemidesmosomal plectin deficiency in a Quarter Horse foal. Equine vet. J. 36: 299-303.

49

Kyaw-Tanner, M & Pollitt C. C. (2004). Equine laminitis: increased transcription of matrix metalloproteinase-2 (MMP-2) occurs during the developmental phase. Equine vet. J. (in press).

French, K.R. & Pollitt, C.C. (2004). Equine laminitis: loss of hemidesmosome ultrastructure correlates to dose in an oligofructose induction model. Equine vet. J. (in press).

French, K.R. & Pollitt, C.C. (2004). Equine laminitis: cleavage of key hemidesmosome proteins associated with basement membrane dysadhesion. Equine vet. J. (in press.)

van Eps, A.W. & Pollitt, C.C. (2004). Equine laminitis: cryotherapy prevents development of the acute lesion. Equine vet. J. (in press).

50

Completed Projects 2005-2006 - Other Project Title

Prototype Environmental Management System (EMS) for the horse keeping community - implementation, auditing and monitoring

RIRDC Project No.:

EPS-1A

Researcher: Ms. Jacqueline Frizenschaf Organisation: Environment Protection Authority (SA) Phone: (08) 8139 9919 Fax: (08) 8139 9901 Email: frizenschaf.jacqueline@saugov.sa.gov.au Objectives

• Phase 1 - Produce a practical prototype EMS framework for the horse

keeping community as a prerequisite and a point of reference for performance-based auditing and development approval.

• Phase 2 - Develop and test a self audit kit for horse keepers through close collaboration between the EPA and the horse keeping communities in different states.

• Phase 3 - Develop an awareness raising and EMS adoption program to reach out to the greater horse keeping community, in the Mt. Lofty Ranges watershed (incl. and interstate component).

Background The project titled ‘Prototype Environmental Management System (EMS) for

the Horse Keeping Community – Implementation, Auditing and Monitoring’ was a 3-year RIRDC-funded research project (referred to as ‘Horse EMS project’ in this report). The project explored the value of an environmental partnership approach between South Australian (SA) horse industry representatives and government to achieve environmental improvements on horse properties on a voluntary basis.

Research The project methodology included a step-by-step design towards the final outcomes, especially the development of a self-audit kit to be promoted to the horse keeping community using a targeted outreach approach. The applied framework used throughout the project was modelled according to the basic elements of International Standardisation Organisation (ISO) 14001’s EMS methodology. This provide a systematic approach to capture the potential risks of horse management activities to the environment; to prioritise action to avoid or mitigate these risks; and to monitor the success of the on-property action(s) with the implied option of continually improving on-property land management.

To achieve the project objectives, the overall methodology was strongly based on a partnership and consultative approach during each project phase. The primary project partners included the EPA and Horse SA, supported by other government and non-government entities. Principles commonly known as ‘participatory development (or management)’ were used to ensure that the project products were endorsed by all parties potentially affected or involved in the future use of the products. A pilot group, comprised of a cross section of the horse industry sector in South Australia was tasked to assist in the development, testing and review of the products. To ensure endorsement from the horse property owners as well as government, environmental land management principles (to be promoted via the products) were reviewed by scientific and government experts (to ensure a sound scientific basis), and tested by the pilot group (to ensure their likely adoption).

The project was rolled out in the Mount Lofty Ranges Watershed, the primary drinking water supply area for metropolitan Adelaide. Testing such a voluntary management effort in an environmentally sensitive region

51

provided a sense of relevance to the project outcomes.

Additional, underpinning tools used as part of the project included: the use of workshop and review sessions; field days; monitoring performance and evaluation for both the desired project outcomes and the actual environmental improvements on the ground. Qualitative and semi-quantitative evaluation tools comprised pilot participant surveys at the beginning and end of project; feedback surveys at the end of workshops and field days; participant reports /presentations on their on farm improvements; mid-term project review with project team.

Throughout the three years or research, the project made extensive use of promotional opportunities such as media releases, and presentations at national and international conferences and local events.

Outcomes • Phase 1 – Prototype EMS developed as a basis for developing a

self-assessment tool for horse keepers. For the first time in Australia, a comprehensive register was compiled which included the most common horse keeping activities and their potential environmental impacts. This list was a valuable basis for a focused approach in Phase 2. The development of the register resulted in a mutual understanding of the fundamental environmental issues potentially caused by horse keeping activities. A total of three workshops, two field days and 10 site visits were held.

Additional products. During Phase 1 workshops and field days a significant knowledge gap of the pilot group (and hence, the larger horse keeping community) became apparent. There was no comprehensive understanding about how the biophysical conditions of a horse property related to the potential enforced detriment of the natural conditions if not managed right. As a result, a Step-by-Step Guide for Land Capability Assessment was devised for the pilot group using in kind support by the AMLR NRMB’s1 Land Management Program. The document guides the user through the process of assessing the biophysical conditions of the property and provides instructions on how to determine environmentally sustainable stocking rates on the horse property, appropriate locations for stables, yards and other intensive horse keeping structures and property maintenance requirements. In addition, a fence post sign was provided to each pilot group participant.

• Phase 2 – Self-assessment tool (now called “Action Planner”) developed. The Action Planner was developed based on a series of workshops and review sessions identifying a pragmatic and user-friendly organisation and structure of the document and its scientific content. It was created to enable the horse keeping community to conduct a quick (about 20-minute long) assessment of how their property management fares. The picture-free, straightforward Action Planner has been tested interstate and received enthusiastic responses. A total of one workshop, one field day and two expert review sessions were held.

Additional products. While drafting the Action Planner, it became evident that enticing horse owners to adopt sound environmental practices would be greatly enhanced if a “how to” guide was added to the package. The Management Guidelines, now accompanying the Action Planner, were developed as a result. Supplementary funding allowed the professional production of this document, which parallels the structure of the Action Planner and contains supporting colour photographs. In addition, one environmental horse poster and several (conference) Horse EMS banners were produced by the EPA and Horse SA.

1 AMLR NRMB – Adelaide & Mount Lofty Ranges Natural Resources Management Board

52

• Phase 3 – Adoption and outreach strategy developed. A well thought-through adoption and outreach strategy was considered an essential element of the project. Such a strategy would allow Horse SA, the custodian of future activities on environmental management by horse keepers, to target its efforts strategically with the goal of ultimately raising the (environmental) performance of the horse keeping community. The strategy was devised based on substantial consultation with both government and the horse keeping community. It now contains a prioritised sequence of strategic actions that provide the highest potential for on-ground environmental improvement results. One workshop and two expert review sessions were held.

Additional products. The project team soon realised that the development of an adoption and outreach strategy would benefit from expert input. Additional NHT2 (Envirofund) funding enabled the team to engage Porter Novelli Pty Ltd in the development of the strategy. Porter Novelli used tools such as workshops and surveys to identify different target groups for an outreach program. In addition, the strategy is underpinned by the principles of community-based social marketing as promoted by Dr. Doug McKenzie-Mohr3.

Implications Although all initially specified project objectives and desired products were

achieved, the overall project success truly lies in the successful application of the participatory development and management approaches used throughout the project. Environmental management for horse keepers won’t wind down after this project, but will carry on into the future, with the following new initiatives established: • community-of-practice forum4; • adoption and outreach monitoring; • periodic updating of Action Planner and Management Guidelines; and • extension of training and research to other ‘lifestyle’ rural land

managers, which often manage horses on their properties.

Publications This project enabled the publication of the following free information resources (available from the Horse SA web site starting in August 2006): • Community-of-practice website (maintained by Horse SA), displaying:

• The Action Planner for Horse Keepers - a tool to assist horse keeper assess and improve environmental management

• Management Guidelines – to accompany the Action Planner for horse properties.

• Frizenschaf J. (2007). Environmental Management on Horse

Properties. RIRDC Publication No 06/137.

2 NHT – Natural Heritage Fund 3 Dr. Doug McKenzie Mohr, 2006. Community-Based Social Marketing (http://www.cbsm.com) 4 Community-of-Practice (CoP) - "builds an effective network which shares knowledge, ideas, expertise and experience regarding content management so as to achieve better practice, improve awareness and understanding, and facilitate successful outcomes (for government)" (Australian Government, Dept. of Finance and Administration). CoPs are often implemented via online learning and exchange forums.