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RIRDC Completed Projects in 2006-2007 & Research in Progress as at June 2007

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To Horses Completed Projects
2.4 HORSES - RESEARCH IN PROGRESS in 2006-2007
PROJECT No PROJECT TITLE RESEARCHER PHONE ORGANISATION
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
AVS-1A Short term and future athletic performance of critically ill equine neonates Dr Jane Axon (02) 6545 1433 Axon Veterinary Services
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
UQ-121A DNA typing of urine samples to confirm the donor identity Dr Ann Trezise (07) 3365 3647 The University of Queensland
UQ-125J Investigation of causal factors for equine amnionitis and foetal loss Dr Judy Cawdell-Smith (07) 5460 1166 The University of Queensland
UQ-132A Pasture management guidelines to minimise risk of laminitis in horses Dr Christopher Pollitt (07) 3365 2063 The University of Queensland
SVH-2J Treatment of placentitis in thoroughbred mares: efficacy of altrenogest Dr Joan Carrick (02) 6545 1333 Scone Veterinary Hospital
UWA-101A Neuromuscular stimulation to reduce joint loading in the equine forelimb Dr Rochelle Nicholls (08) 9431 3862 University of Western Australia
UQ-118A Laminitis preventive and therapeutic strategies Dr. Christopher Pollitt (07) 3365 2063 The University of Queensland
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
UQ-129A Identification of horses with resistance to small strongyle infections Dr Glen Coleman (07) 3365 2119 The University of Queensland
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
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
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
UF-11A SHARE - Safety for horses and riders in eventing Dr Raymond Cripps (08) 8201 7627 Flinders University of South Australia
UM-58A Epidemiology and risk factor analysis of racetrack fatalities Prof. Andrew Clarke (03) 9731 2314 The University of Melbourne
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
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
RWE-2A Tracheal stethoscopy: An effective tool for diagnosing DDSP and evaluating treatment? Mr Robert Curtis (02) 4757 4876 Randwick Equine Centre

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 ·1 Identify the major mechanisms of chronic lung lesions induced by EIPH.

·2 Develop methods for reducing chronic ling damage by modulating macrophage-related events after episodes of EIPH.

·3 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.

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 ·4 To determine admission parameters and clinical problems that will affect the survival of the critically-ill neonatal foal.

·5 To determine whether surviving foals reach their sale potential as yearlings. 

·6 To determine whether surviving foals are able to have one race. 

·7 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.

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 ·8 To identify the magnitude and direction of motion available in both clinically normal and diseased equine sacoiliac joints

·9 Correlate clinical signs, force plate analysis and three dimensional kinematics in horses affected by sacroiliac disease

·10 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: ·11 the greatest relative movements in the sacroiliac joint complex are in the transverse and coronal planes

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

·13 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.

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 ·1 Test various methods for DNA typing of horse urine samples.

·2 Establish and optimise a method that is reliable for drug-test positive samples.

·3 Validate and refine the procedure for routine operation.

·4 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.

 

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 ·5 To determine whether administration of processionary caterpillars (Ochrogaster lunifer) to pregnant mares is associated with increased risk of foetal loss and abortion.

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

·7 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.

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 ·8 Investigate and raise awareness of the problems of excess carbohydrate in forage as a risk factor in laminitis

·9 Determine levels of NSC in grass that triggers laminitis

·10 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. 

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: ·11 Antibiotic and anti-inflammatory drugs (Trimethoprim-sulphadimidine and flunixin meglumine)

·12 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.

 

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

·2 Evaluate the effectiveness of the sleeve in reduction of joint loads during gait

·3 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
 
 

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.





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 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: 

·1 radial carpal extensor m.

·2 common digital extensor m.

·3 extensor carpi ulnaris m.

·4 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:

·5 without sleeves (baseline);

·6 with sleeves but nil vibration;

·7 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.

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

·5 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.

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. 

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.

 

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 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.

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 ·1 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.

 

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.

 

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.

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 ·6 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. 

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:

•1 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

•2 formalised monitoring of injuries and fatalities

•3 standardisation of track rating methods and review of current policy regarding racing on ‘good’ tracks

•4 further investigation of differences between risk in hurdles and steeplechase races

•5 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.

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.

 

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 ·1 To identify the incidence of radiographic lesions in yearling thoroughbred horses presented for sale in Australia.

·2 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.

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. 

 

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: