YTCAF GRANTS SUPPORT --- 2010

INFORMATION ON GRANT SELECTION PROCESS

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Each year the AKC Canine Health Foundation send the YTCA Foundation a list of grant requests for funding that they have screened and think would be of interest to breeders and owners of Yorkshire Terriers as well as dog fanciers in general.  As an independent entity spun off from the Parent club to preserve our tax exempt status, the YTCAF receives no financial support from the YTCA's participating in efforts such as the Purina Circles program.  Instead all of our funding comes from memorials made by individual donors and fund raising efforts such as the online auctions and breeding calendar sales.  There are currently no matching funds available to increase the size of our donations.  Thus our research budget for the remainder of 2010 was limited to $7,000.

In August 2009 fourteen grants were submitted to the YTCA Foundation for consideration.  The Board members carefully considered these factors in each of the requests in order to optimize research dollars:

1. 1. Potential of the research to benefit Yorkshire Terriers in particular and all dogs in   general.

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3. 2. Previous research success of the primary investigators

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5. 3.  Use of the grant project to help veterinary students, new  veterinarians and academicians get started in their chosen field of research (Acorn projects).

After a 2-month investigative process the Board voted to support these CHF grants for 2010.

CHF Grant Funding

CHF Grant 1352-A --- $500

Iowa State University - 2010

Detection of Brucella canis DNA in Canine Urine, Semen and Vaginal cells via qPCR Analysis

Lin Kauffman, DVM

CHF Grant 1352-A --- $500

Iowa State University - 2010

Detection of Brucella canis DNA in Canine Urine, Semen and Vaginal cells via qPCR Analysis

Lin Kauffman, DVM

Project Abstract

Brucella canis is a bacteria that causes reproductive disorders in dogs.  The number of B. canis cases has been on the increase across the US and specifically the Midwest over the past several years with increasing costs associated with reproduction losses due to disease and euthanasia of infected animals.  Prevalence in Oklahoma alone has risen from 2% in 1994-95 to 13% in 2002-03.  The potential for human infection from dogs increases concern over this disease.  Currently the only available tests for this disease are microbiological cultures and serological testing.  Microbiological culture of the blood, vaginal secretions, semen or fetal placenta is the diagnostic gold standard since a positive culture confirms the disease.  Unfortunately the organism is hard to culture under the best conditions.  Serological tests detect the disease 8-12 weeks postinfection and cannot detect early disease, from the time of infection until when there are detectable levels of anti-Brucella antibodies.  At present, laws make canine brucellosis a reportable disease subject to quarantine.  In some states this mean testing all breeding animals and euthanasia of infected animals.  The economical impact of quarantine and repeated testing is such that there is potential for individuals to have their dogs tested out of state to avoid quarantine, sell known infected animals and/or to medically treat potentially infected animals to avoid detection with current tests.  Brucella canis has infection potential to humans, especially the very young, elderly and immunocompromised individuals.  Currently there is no early detection testing available for this disease.  Early detection of disease would shorten the quarantine period and would add a measure of safely for buyers of puppies or breeding stock.  The goal of this project is to use the Polymerase Chain Reaction (qPCR) based assay to detect Brucella sp. in field samples of suspect B. canis urine (male), semen and vaginal cells and compare how well this test works verses traditional culture and serology.

UPDATE 7-31-10

Brucella canis, a bacteria that causes reproductive disorders in dogs, has been on the increase across the US.  This goes hand in hand with increasing costs associated with reproduction losses due to disease and euthanasia of infected animals.  The potential for human infection from dogs increases concern over this infectious disease.  Currently the only available tests for this disease are not great.  The bacteria is hard to culture and serological tests detect the disease 8-12 weeks post-infection but cannot detect early disease.  Present laws make canine brucellosis a reportable disease subject to quarantine.  In some states this means testing all breeding animals and euthanasia of infected animals.  Early detection of disease could shorten the quarantine period and would add a measure of safety for buyers of puppies or breeding stock.  The goal of this project is to use Polymerase Chain Reaction (qPCR) assay to detect Brucella sp. in samples of suspect B. canis urine (male), semen and vaginal cells (female) and compare how this test works vs. traditional culture and serology.

To find a more specific and timely diagnostic, this study assessed the ability of qPCR analysis to detect B. canis Omp25 DNA in a variety of samples (blood, urine, vaginal swab) and compared those results against current detection methods for B. canis infection in dogs (serology). qPCR analysis identified the presence of B. canis Omp25 DNA in multiple dogs prior to seroconversion.  Non-invasive samples from the genito-urinary tract, including vaginal swabs and urine, were found to be the most sensitive for detection of B. canis Omp25 DNA via qPCR.  Use of these samples would make collection of diagnostic samples within the ability of some dog owners and breeders.

The results of this study are very encouraging for use of B. canis Omp25-specific qPCR as a diagnostic screening tool for B. canis.  The potential of this assay qPCR for early detection could be very valuable for elimination of B. canis from kennels without having to wait for seroconversion.  Additionally, Omp25 qPCR could be a valuable screening tool for B. canis in newly purchased dogs prior to adding these dogs into a new home or kennel.  B. canis is a reemerging infectious disease in the canine breeding industry.   A better screening and detection method will be very useful to prevent further spread of this insidious disease.  B. canis Omp25 qPCR may be this critical diagnostic component to decrease economic effects of canine brucellosis on the canine breeding industry and prevalence of canine brucellosis in the US.

  Board's Assessment

The bacterium Brucella canis causes major canine reproductive problems, and the incidence has been on the rise in recent years.  It is a major concern to both breeders and purchasers of dogs because current tests to detect its presence take between 2 and 3 months to complete.  Concurrently brucellosis is a zoonosis and spreads easily to humans.  For this reason it is a reportable disease and affects dogs have to be either quarantined or euthanized.  Dr. Kauffman is seeking to use an assay based on the Polymerase Chain Reaction (qPCR) technique (which is very fast) as a detection tool and compare these results with standard test results to see if this new technology would be a more accurate and faster method of testing dogs for the presence of B. canis.  Previously Dr. Kauffman has been a featured speaker on the CHF Genome Barks podcasts.

Amount Funded by YTCAF = $500.  Rationale for support of this grant:

The YTCAF voted to support this research because of its importance to all dogs including Yorkshire Terriers.  Breeders are aware of the disastrous consequences of having breeding stock infected with B. canis.  Additionally this bacterium can cause serious illness in humans.

 CHF Grant 1105 --- $2,500

University of Florida - 2010

Understanding the Dynamics of Canine Influenza Virus Transmission in Dog Populations and Intervention Strategies for Reducing Transmission

Cynda Crawford, DVM, PhD

  Board's Assessment

The new H3N8 Canine Influenza Virus (CIV) was first observed in greyhound kennels several years ago.  It is thought to be a mutation of a horse virus.  Since then there have been sporadic outbreaks.  While the majority of dogs survive the viral infection, a small proportion die as a result of the subsequent pneumonia.  The purpose of this study is to map out the epidemiology of this new virus; how is it transmitted, what parts of the canine population are at greatest risk, and what treatments have been most effective.  A compute model will then be created and used to predict what efforts will be most successful at stopping the spread of the Virus and reducing the opportunities for transmission between dogs.  This model could then be used by planners of dog events, boarding/show kennels, etc.  Dr. Crawford has been studying this virus for the past several years in Florida and currently holds the provisional patent on a new vaccine against H3N8.  She hold a PhD in Immunology/Infectious Diseases as well as a DVM.

Amount Funded by YTCAF = $2,500.  Rational for support of this grant:

The YTCAF voted to support Dr. Crawford's research because of its direct implications for the planning of all competitive AKC dog events, as well as individual show kennels.  Recent outbreaks of H3N8 in New Jersey and Northern Virginia have highlighted the need to know how this virus is spread and which dogs are at greatest risk.

CHF Grant 1341-A --- $2,000

Cornell University - 2010

Sample Collection, Pedigree Analysis and Candidate Gene Screen for Protein-Losing Enteropathy in Yorkshire Terriers

Nate Sutter, PhD

CHF Grant 1341-A ---$2,000

Cornell University - 2010

Sample Collection, Pedigree Analysis and Candidate Gene Screen for Protein-Losing Enteropathy in Yorkshire Terriers

Nate Sutter, PhD

Project Abstract

Protein-losing enteropathy (PLE) is a life threatening condition diagnosed in many breeds including Soft Coated Wheaten Terrier, Basenji, Lundehund, Chinese Sharpei and Yorkshire Terrier.  The syndrome is particularly prevalent in Yorkshire Terriers.  These dogs commonly suffer from chronic diarrhea and lose protein into the intestine.  A variety of abnormalities in the intestinal lining are thought to be involved.  While dietary changes sometimes help, treatment is supportive rather than curative.  As many as half or more of all Yorkshire Terriers diagnosed with PLE ultimately succumb to it.  We want to identify the genetic risk factor or factors that predispose Yorkshire Terriers to this syndrome.  These genetic factors may also be present in other breeds, a hypothesis that can be tested once the factors are identified in Yorkshire Terriers.  Here, we propose to collect blood samples for DNA, pedigrees, and clinical data from affected and unaffected Yorkshire Terriers.  We will analyze the pedigrees for patterns of coancestry.  We will also sequence a gene, syndecan l, that is a key player in PLE in people, as we hypothesize it plays a role in canine PLE.

UPDATE 5-31-10

Yorkshire Terriers and other purebred dogs suffer from a debilitating protein-losing enteropathy (PLE) syndrome of uncertain causes.  The disease presentation frequently includes diarrhea, vomiting, inappetance, panhypoproteinemia, and weight loss.  Intestinal histopathology is dominated by lymphangiectasia.  Recently, crypt 'cysts" containing proteinaceous material have been described (Kenneth Simpson, personal communication), PLE is frequently fatal in Yorkshire Terriers, and the complexity of presenting complaints, as well as the variable age of onset, suggests the syndrome may be under-diagnosed.  The relative risk for Yorkshire Terriers is above 10 and suggests that one or several heritable components predispose members of this breed to PLE. 

We continue to recruit PLE affected and older unaffected Yorkshire Terriers for a pedigree analysis of co-ancestry among affected dogs, DNA collection and medical data analysis.  To date, we have collected 66 out of 100 samples.  The candidate gene screen of syndecan 1 proposed in this project has so far found no evidence that sequence variation at this gene associated with PLE in the Yorkshire Terrier.

  Board's Assessment

Of the many breeds of dogs that are diagnosed with Protein-Losing Enteropathy (PLE), Yorkshire Terriers are among the most affected.  This condition is characterized by chronic debilitating diarrhea along with the loss of protein from the body  into the intestinal tract where it is passed out in the feces.  There is currently no cure and individual dogs can only be clinically helped with supportive therapy through the use of drugs and special diets.   Because of the high number of Yorkshire Terriers afflicted with PLE, Dr. Sutter will try to establish a genetic link through the collection of blood samples and pedigrees from dogs in our breed.  This information could then be extrapolated for other breeds that are similarly affected.  Dr. Sutter has been studying canine genetics for some time, Particularly the mutations that make dogs susceptible to cancer and other diseases.  Previously he was a Postdoctoral Fellow at the Fred Hutchinson Cancer Research Center and a Research Fellow at the National Human Genome Research Institute at NIH.

Amount Funded by YTCAF = $2,000.  Rationale for support of this grant:

Almost 50% of Yorkshire Terriers who have PLE die from it.  Currently there is no screening test for this disease that is suspected to have a genetic basis.  The Foundation's support of this grant will hopefully pave the way for the identification of the gene(s) responsible for PLE and the creation of a test to screen breeding stock as well as individual Yorkies for it.

CHF 1160-A ---$1,000

University of Pennsylvania - 2010

Development of Contract-Enhanced Magnetic Resonance Angiography for the Diagnosis of Congenital Vascular Liver Disease in Dogs

Wilfried Mai, DVM, PhD

CHF 1160-A ---$1,000

University of Pennsylvania - 2010

Development of Contract-Enhanced Magnetic Resonance Angiography for the Diagnosis of Congenital Vascular Liver Disease in Dogs

Wilfried Mai, DVM, PhD

Project Abstract

Porto-systemic shunts (PSS) are anomalies of the bessels to the liver (abnormal communication between the portal vein, which feeds the liver, and another veine.g. (vena cava).  This results in blood from the gut not being detoxified by the liver causing clinical signs (seizures, stunted growth, fluid accumulation in the abdomen...) and ultimately death, if untreated.  Most PSS are congenital.  Several breeds are predisposed and some hereditary components have been documented (e.g. Maltese, Yorkshire Terriers, Irish Wolfhound, Australian Cattle Dogs, Golden and Labrador Retrievers).  Diagnostic imaging techniques such as X-rays, ultrasonography, and scintigraphy are currently used to diagnose this condition, and are crucial not only to diagnose it but also to decide what the best treatment option will be.  None of these techniques is perfect and all have major disadvantages so that often, several techniques must be used in conjunction to get the information needed.   magnetic resonance imaging can image blood vessels with the use of a dye --- a technique known as contract-enhanced magnetic resonance angiography (CE-MRA) --- and is now routinely used in people to diagnose many diseases of the vessels.  This technique overcomes many of the inconveniences of the other techniques and our goal is to develop this method to diagnose congenital porto-systemic shunts in dogs.

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NOTE:  Dr. Mai's grant began with CHF 5-01-2008 and is expected to end 4-30-10.

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Update 10-26-2009

The proposed research of utilizing contrast-enhanced portal magnetic resonance angiography (CE/MRA) in dogs to diagnose congenital liver vascular diseases was applied to 15 dogs so far.  In all of them, the technique was easily and successfully performed, and yielded good to excellent quality images in less than 4 minutes total imaging time, which is much shorter than the current method used (single-slice helical computed tomography angiography).  This in turn shortens total anesthesia time for diagnostic imaging in comparison with computed tomography.  CEMRA provided diagnostic information in all 15 dogs, with excellent contrast and anatomic detail.  No complications were observed.  The three-dimensional images obtained are easily understood by the surgeons in charge of the patients.  In all 11 animals that underwent treatment through coil embolization or surgery to close off the anomalous vessel(s), the anomalies seen on CE/MRA were confirmed.  Four of the 15 dogs have not yet undergone this procedure.  The method is very accurate in terms of anatomic depiction of the vascular abnormalities.  12 dogs were diagosed with intra-hepatic  congenital porto-caval shunts, two with an extrahepatic shunt and one with a rare form of intra-hepatic arterio-venous fistula.  The preliminary results were presented at the Annual Conference of the American College of Veterinary Radiology, where they impressed the audience because of the ease and simplicity of the method, its non-invasive nature and the high quality and diagnostic value of the images obtained.  Two very small puppies were imaged and despite the challenges associated with imaging such small patients, the studies were diagnostic in both cases and anesthesia well tolerated with uneventful recovery.  In two animals imaged, ultrasonography prior to the MRA by experienced radiologists failed to accurate describe the anatomy of the shunt or to confirm its presence.

UPDATE 4-30-10

The proposed research of utilizing contrast-enhanced portal magnetic resonance angiography (CE-MRA) in dogs to diagnose congenital liver vascular diseases was applied to 17 dogs as planned.  In all of them, the technique was easily and successfully performed, and yielded good to excellent quality images in less than 4 minutes total imaging time, which is much shorter than the current method used (singled-slice helical computed tomography angiography).  This in turns shortens total anesthesia time for diagnostic imaging in comparison with computed tomography.  CE-MRA provided diagnostic information in all 17 dogs, with excellent contrast and anatomic detail.  No complications were observed.  The three-dimensional images obtained are easily understood by the surgeons in charge of the patients.  In all 14 animals that underwent treatment through coil embolization or surgery to close-off the anomalous vessel(s), the anomalies seen on CE-MRA were confirmed.  Two of the 17 dogs have not yet undergone this procedure.  The method is very accurate in terms of anatomic depiction of the vascular abnormalities.  13 dogs were diagnosed with intra-hepatic congenital porto-caval shunts, two with an extra-hepatic shunt and one with a rare form of intra-hepatic arterio-venous fistula.  one dog did not have a macroscopic anomalous vessel identified, consistent with hepatic microvascular dysplasia.  The preliminary results were presented at the Annual Conference of the American College of Veterinary Radiology, the British Small Animal Veterinary Association Conference and the European Association of Veterinary Diagnostic Imaging British/Irish Division Conference, where they impressed the audience because of the ease and simplicity of the method, its non-invasive nature and the high-quality and diagnostic value of the images obtained.  Two very small puppies were imaged and despite the challenges associated with imaging such small patients, the studies were diagnostic in both cases and anesthesia well tolerated with uneventful recovery.  In two animals imaged, ultrasonography prior to the MRA by experienced radiologists failed to accurate describe the anatomy of the shunt or to confirm its presence.

Board's Assessment

Dr. Mai is seeking to modify the technique of using contract-enhanced magnetic resonance angiography (CE-MRA) in humans for use in dogs, particularly those breeds which are known to have abnormalities of the blood vessels (i.e. Vasculature) in the liver.  Traditional methods for diagnosing porto-systemic shunts (PSS) and hepatic microvascular disease (MVD) have included radiographs (X-rays), ultrasound, and other tests like scintigraphy.  These tests often have to be use in combination to obtain an accurate picture of the extent of the problem before treatment options can be obtained.  Hopefully CE-MRA will afford a better diagnostic option for canine liver patients.  Breeds that are particularly affected include Yorkshire Terriers.  Dr. Mai holds a DVM as well as a PhD in Biomedical Engineering/Core Medical Imaging.  He has extensive research experience in the use of different radiology techniques in veterinary medicine.

Amount Funded by YTCAF = $1,000.  Rationale for support of this grant:

This technique may prove to be both superior and less invasive than currently used methods of diagnosis for canine liver disease.  This is important because accurate diagnosis often makes the difference between life and death for these dogs.  Since there is a high prevalence of Yorkshire Terriers affected by either PSS or MVD, the Foundation thought that it was important to support Dr. Mai's research.

CHF 1241-A --- $500

National University of La Plata - 2010

Detection of DNA Damage in Response to Cooling Storage in Canine Spermatozoa Using Single-cell Gel Electrophoresis (Comet Assay)

Cristina Gobello, DVM