Australian National Kennel Council Limited

Abstracts from Canine Research Foundation Programs

The Canine Research Foundation (CRF) is the official research vehicle of ANKC Ltd. It is a Public Charitable Trust with research conducted at Australian universities from funds generated via the levy on puppy registrations, fund raising functions, tax-deductible donations and bequests from the public. The CRF was founded by the VCA in 1992 and 70 research grants have been awarded for projects through to those commencing in 2008. Funding is now flowing to CRF from  most ANKC Member Bodies, and excluding CCCQ which has a separate research program in conjunction with the University of Queensland.

Summaries of more recent research projects are included below to assist owners and breeders in becoming aware of the significant results being achieved in canine research directed at improvement of canine health, and as an important Australian contribution on the world scene. Most projects to date have been successful, with results published in veterinary/scientific journals and web-sites available to veterinarians and the canine community internationally. We can all be justifiably proud of results.

Breed clubs/councils are invited to nominate research topics and forward them to the ANKC Administrator for consideration by January 31 each year.

Recent Research Grants
The following brief summaries cover CRF grant awards and progress to date for research projects conducted in 2002-2006.

1. Comparison of chemiluminescence and radioimmunoassay methods for estimating canine blood cortisol levels
(Investigator: Dr. P.Irwin: Veterinary Clinical Science, Murdoch University)

Serum cortisol values are used to determine whether dogs have significant adrenal axis illness, current adrenal function, and to monitor therapy for adrenal illness. Absolute serum cortisol values are crucial when determining whether to initiate or proceed to therapy for adrenal disease in dogs. The RIA method is the accepted standard, but chemiluminescence is the method used in most commercial assays. Experience has shown serious variation in results between the two methods.

Research findings are that although RIA and chemiluminescence cortisol concentrations appear highly correlated, a significant difference may exist when measuring cortisol concentrations less than 100nmol/L in stored canine sera. The results of chemiluminescence cortisol assays should be interpreted with caution unless the specific assay method in the laboratory has been adequately validated in normal dogs and dogs with hyperadrenocorticism.
(Final report received and also submitted for publication to the American College of Veterinary Internal Medicine.)

2. Disease Gene Mapping
(Investigator: Dr A.Wilton: University of NSW)

Ceroid lipofuscinosis (NCL) in Border Collies has been the main focus of this research over some years. The funds from CRF grants, the Batten’s Disease Association and from Border Collie Clubs allowed attraction of extra funding from a government  SPIRT grant.

Development of the canine genome map allowed a change in approach to the identification of the NCL gene.  Regions of conservation between dog and human gene
maps and the knowledge of 7 different genes that can cause similar symptoms in humans provided a good starting point to identify candidates for the NCL gene.  Genetic markers (microsatellites) close to the presumed site of these candidate genes were typed in the available Border Collie NCL-affected pedigrees to indicate whether one of the homologues of the human NCL genes is a likely cause of NCL in Borders. Work was also conducted on NCL in collaboration with the laboratory of Frode Lingaas in the Norwegian Veterinary School where they work on CL in English setters. Preliminary data suggested that Border Collie NCL and English Setter CL were the same gene on chromosome CFA37, but this was not supported by the additional pedigrees and markers tested.

The causative gene for Border Collie NCL now has been identified. It is the same mutation as one causing Batten’s disease in humans. A direct genetic test for Border Collie NCL is now available.
(Project completed, final report received and published in 'Genomics'.)

3. Prevention of uterine infection, cystic endometrial hyperplasia and pyometra
(Investigator: Dr P.Wright, University of Melbourne)

This project follows on from results of a major CRF-funded study “Uterine function and infertility in the bitch.” Three papers arising out of this original work were invited for publication by the 4th. International Symposium on Canine and Feline Reproduction held in Oslo.

A simple procedure for the induction of whelping was developed to assist bitches with whelping problems, and, where difficulties may be expected, at a time when optimal assistance is available. This involves the administration of the drug prostaglandin. Whelping commences around 36 hours later.

Cystic endometrial hyperplasia involves the formation of cysts in the lining of the uterus. This is a common cause of infertility. The condition predisposes the bitch to uterine infection. This can lead to a life-threatening condition called pyometra. This is a severe uterine infection with the accumulation of pus in the uterus. Research is now on development of a vaccine to prevent pyometra.
(Funding completed, awaiting final report.)

4. Effect of oral ursodeoxycholic acid on bile acid tolerance tests in healthy dogs
(Investigator: Dr L.Abraham: Veterinary Science, University of Melbourne)

Significant hepatic disease is characterised by normal or elevated preprandial (before feeding) and elevated postprandial (after feeding) serum bile acid (SBA) concentrations. Although SBA values cannot differentiate between liver diseases, elevated SBA values may be seen with such disorders as cirrhosis, chronic hepatitis, hepatic necrosis, neoplasia, portosystemic vascular anomalies and cholestasis. Bile acids are synthesised from cholesterol exclusively in the liver and measurement of SBA concentration is routinely used as a diagnostic test for liver function. The bile acid tolerance test consists of oral administration of bile acids with measurement of SBA concentrations to monitor liver function in maintaining SBA levels.

The major bile acids are cytotoxic at sustained elevated levels, possibly leading to complete hepatic failure. Ursodeoxycholic acid (UDCA) is a bile acid usually present in trace amounts only and is not cytotoxic in many species. In human patients, UDCA has been used to treat various chronic cholestatic disorders, but there are few reports of its use in dogs and no clinical trials have been reported. UDCA decreases the cytotoxicity of
bile acids by decreasing the proportion of other bile acids and increasing the proportion of UDCA synthesised and secreted by hepatocytes, reducing the risk of liver failure.

Results of this research project show that administration of UDCA does not alter the bile acids tolerance test of normal healthy dogs, and it would appear that the bile acids tolerance test can be used to monitor hepatic function in dogs with liver disorders that are being medicated with UDCA.
(Project completed, final report received.)

5. Improving therapeutic control of seizures in dogs
(Investigator: Dr M.Govendir, Veterinary Science, University of Sydney)

Accepted therapy for minimizing the frequency and severity of seizures in dogs with idiopathic epilepsy (IE) is daily administration of phenobarbitone (PB) and/or potassium bromide (KBr). These drugs are generally effective, affordable and allow dosing once or twice daily. However approximately 20-50% of epileptic dogs treated with PB alone will continue to have unacceptable seizure activity and will require combination therapy incorporating KBr. Even then a proportion of dogs still have significant seizure activity.

This study was initiated to determine whether dogs with refractory IE would attain improved seizure control when gabapentin (GP) was added to the therapeutic regime.

Research findings were:-

• most dogs treated with PB, KBr and GP either stopped seizuring altogether or    had a reduction in seizure frequency and particularly duration
• GP medication had no major side effects
• one of the side effects of prolonged PB medication is development of liver    toxicity. In many dogs that have been on long term PB treatment, withdrawal of    PB may result in an increase in seizure activity. Treatment with GP prevented    PB withdrawal seizures and allowed PB to be completely withdrawn.
• the majority of patients medicated with PB for a long period had significant    fasting hypertriglyceridaemia.

Further research is indicated on the long term use of PB and the prospect of medication with GP alone.
(Project completed. Final report received. See project 11 below for further research.)

6. Analysis of Major Histocompatability Complex II type in dogs with granulomatous meningoencephalitis
(Investigators: Drs. S.Holloway & L.Abraham: University of Melbourne Veterinary Science,)

The aim is to assess the role of MHC II type molecules in the pathogenesis of GME in dogs. GME is an inflammatory disease of the central nervous system. It is the most common neurological disease of small breed dogs. It is usually fatal. The hypothesis is that GME is an autoimmune disease due to a type IV T-cell mediated hyposensitivity, and that inheritance of specific type II MHC molecules predisposes dogs to develop GME autoimmunity.
Primers have been identified and tested for amplifying the hypervariable areas of MHC II genes located in exon 2 of the two MHC loci DLA-DRB1, and DLA-DQB. Work is proceeding to obtain a reliable PCR result on small quantities of genomic DNA.
(Funding completed, project due for completion late 2006.)

7. Liver dysfunction in Tibetan Spaniels
(Investigator: Dr. L.Abraham: Veterinary Science, University of Melbourne)

The aim is to investigate the cause and possible mode of inheritance of abnormal results of bile acid tolerance (liver function) testing in Tibetan Spaniels.

Although many disorders can influence serum bile acid concentrations, hepatic disease is characterised by normal or elevated serum bile acid concentrations. Although serum bile acid values cannot differentiate between liver diseases, elevated serum bile acid values may be seen with diseases such as cirrhosis, chronic hepatitis, hepatic necrosis, neoplasia, portosystemic vascular anomalies and cholestasis. Bile acids tolerance testing is used commonly as a screening test for identifying young dogs with portosystemic vascular anomalies. There is evidence from the Maltese breed that an abnormal bile acids tolerance test result does not necessarily reflect abnormal liver function for some breeds and this study is aimed at determining whether the bile acids tolerance test is an appropriate test of liver function in the Tibetan Spaniel. In addition, the study aims to determine the type of liver disease that may be present in this breed. Results may have a wider application to other breeds.
(Funding completed, awaiting final report.)

8. Role of toxigenic Pasteurella multocida in the development of atrophic rhinitis
(Investigator: Dr. J. Sandy: Veterinary Science, University of Melbourne)

The aim is to determine whether toxigenic strains of P. multocida are present in cases of canine atrophic rhinitis, so that a rapid, inexpensive diagnostic test and rational treatment for this condition can be investigated.

Canine rhinitis can be a debilitating disease, especially where destruction of the nasal turbinate structure has occurred. Extensive bone atrophy and remodelling frequently accompanies cases of chronic rhinitis attributable to established nasal pathogens such as Aspergillus and Penicillium species, but coinfection with toxic P. multocida may contribute to the disease associated with these pathogens. Accurate diagnosis of the causes of atrophic rhinitis is warranted to prevent extensive structural damage from occurring.
(Funding completed, awaiting final report.)

9. Use of transcolonic portal scintigraphy to evaluate efficacy of cellophane banding for congenital portosystemic shunts in dogs
(Investigator: Dr L. Abraham, University of Melbourne, Veterinary Science)

The objective of this project is to evaluate portal circulation in dogs following cellophane banding of single congenital intra- or extra- hepatic portosystemic shunts by use of transcolonic portal scintigraphy, and to determine if elevated post-operative cellophane-banded shunt fractions are due to failure of shunt closure (attenuation) or development of multiple acquired shunts using mesenteric portovenography.

The hypothesis is that in dogs with post-operative shunting identified following cellophane banding, the shunting of blood is due to development of multiple acquired shunts, rather than failure of complete attenuation (closure) of the shunt.

Results so far have shown two cases of persistent shunting 10 weeks following cellophane banding of portocaval shunts by use of portal scintography. Both underwent a laparotomy and portovenogram. Both cases were identified with multiple portocaval shunts. This is an important development  - the first world-wide follow up cases of persistent shunting with portovenography. The aim now is to obtain 20-30 cases  for viable statistical evaluation.
(Funding completed, awaiting final report.)

10. Using the dog genome sequence to identify genetic diseases
(Investigator: Dr. A. Wilton: University of NSW)

The aim of the project is to use the information from the dog genome sequence (released in July 2004) to identify genes responsible for genetic diseases in dogs. The dog genome provides information on genes that can be assessed as candidates for known diseases based on gene function and comparative disease data to other species such as human and mouse. DNA sequencing can then be used to determine which of the genes are causative of disease. This approach has been used successfully in the lab to identify the gene for NCL in Border Collies.

Research will now concentrate initially on the disease Trapped Neutrophil Syndrome (TNS), seen in Border Collies, which is an immune dysfunction resulting in multiple infections and failure to thrive in pups.
(Funding suspended)

11. Long term use of phenobarbitone in idiopathic epilepsy
(Investigator:  Dr M Govendir, University of Sydney, Veterinary Science)

This proposal follows on from Dr Govendir’s previous project in which Gabapentin has been shown to be useful in treating seizuring dogs without any side effects. During this prior work it was found that some dogs continued to experience seizures with long term PB treatment and it is considered important to investigate why.

Canine idiopathic epilepsy (IE) is the most common neurological disorder in dogs. Estimates of the incidence range from 2 to 3 % of total hospital admissions. Presently phenobarbitone (PB) is the most commonly used therapeutic agent for medical management of IE. Dogs with IE generally remain on anticonvulsant medication throughout their lives.

Traditionally, veterinarians initially treat canine epileptics with PB. This drug has been the “gold standard” as it minimises the frequency or severity of seizures initially and is associated with acceptable side effects, is relatively inexpensive for life-time medication and there are few other effective canine anticonvulsant agents. Unfortunately, approximately 20-50% of canine epileptics will eventually demonstrate inadequate seizure control on PB even at high doses, and another therapeutic agent such as KBr must be added to the therapeutic regime to improve seizure control.

Progress reports show that not all dogs on anticonvulsant medication have high fasting fat levels, however a small proportion do have significant fasting fat levels - not previously described in veterinary literature as yet. Fasting triglyceride ranges for normal dogs have not been established and consequently this is being done in this project. Fasting triglyceride and cholesterol concentrations may differ for various breeds of dogs and investigation is continuing to identify whether specific breed variation exists.
(In progress, funding continuing.)

12. Human-animal interactions and behavioural problems in dogs
(Professor Paul Hemsworth, University of Melbourne: $10,000 per year for 3 years)

This is part of a large linkage project of the same title proposed to the Australian Research Council. A number of notable research personnel from various research establishments will be involved, along with a number of industry partners. Results should be of great significance in improving the health, well-being and position of dogs in suburbia.

It is estimated that 37% of Australian households have at least one dog. However companion animals can create a number of problems in the general community and problem behaviour in dogs has become a major issue for both dog owners and veterinarians. The most common problems presented to veterinarians are aggression, excessive vocalisation, destructive behaviour, barking and jumping up on people.

Preliminary and limited research results support the contention that the relationship between humans and animals is a key determinant of some behavioural problems in dogs and may impact on dog welfare.

The project will study the effects of both the amount and nature (quality) of human contact on fear, anxiety and the common behavioural problems seen in dogs in suburbia. Behavioural problems in dogs may be true behavioural problems (ie. behavioural disorders) for the animals. True disorders may develop if the owner provides inconsistent or unpredictable reinforcements or the welfare of the animal is severely challenged, Alternatively, some behaviours labelled as abnormal may be normal behaviours in free-living populations but are considered undesirable by humans and/or owners. Thus dog behaviour problems can be considered as behaviours of the dog that owners generally considered a problem, either to themselves, others or the dog.
(In progress, funding continuing,.)

13. Evaluation of abnormal serum bile acids in the Maltese breed
(Dr. Linda Abraham, University of Melbourne,Veterinary Science)

This is a follow on from previous research by Dr. Abraham reported above. The objective is to investigate the cause of abnormal liver function test results in Maltese, which also may be relevant to other breeds

There is evidence from a previous study of the Maltese breed that an abnormal bile acids tolerance result does not necessarily reflect abnormal liver function for this breed. The current study aims to determine the type of liver disease and composition of bile acids that may be present within dogs of this breed with abnormal liver function test results in the absence of a portosystemic shunt. Another aim is to compare pedigrees of the dogs involved to examine the possible mode of inheritance of portosystemic venous anomaly and other causes of elevated serum bile acids.
(In progress, funding continuing.)