Targeted Next Generation Sequencing Panel for Comprehensive Testing for Reproductive and Neurologic Pathogens of Dogs

The large number of pathogens that contribute to infectious reproductive or neurologic disease and overlap of clinical signs makes using tests for multi-pathogen detection useful. Routinely, several individual or multiplex PCR assays are performed to identify active infection with these agents.

Determining which tests to use for any given case can be difficult. Therefore, investigators propose to validate a comprehensive targeted next generation sequencing (NGS) panel for detection of all known neurologic and reproductive pathogens that infect dogs, as well as other infectious pathogens of dogs, including vector-borne agents previously validated through our work funded by AKC CHF. This takes the guesswork out of proper test selection for infectious disease diagnostics.

The amount of data that can be generated with NGS allows for comprehensive testing, and use of a targeted method (adding primers to the protocol for amplification of targets of interest) reduces the cost to the point that this testing can be effectively used in a veterinary diagnostic lab. More and more veterinary laboratories are purchasing sequencing equipment, so this method has merit to be expanded to additional labs. With the comprehensive nature of the panel, epidemiological evaluation of infectious diseases in the dog population can also be improved.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 03137

RESEARCHERS

Rebecca Wilkes, DVM, PhD; Purdue University
Amount: $3,000

Characterization of hookworm resistance in dogs with a novel diagnostic test for early intervention

The dog hookworm is the most common worm parasite of dogs in the United States. It can cause disease, resulting in suffering and even death in both young and old dogs of all breeds. Dog hookworms can also infect humans who come into contact with hookworm larvae present in dog stool contaminated areas. Hookworm infections in people will cause cutaneous larval migrans, a painful skin condition. Dogs must be dewormed monthly to prevent the ill effects of acquiring hookworms. While these medications were very effective 20-30 years ago, recently they have stopped being fully effective in many cases. This is because hookworms have become resistant.

To know if a dog has resistant hookworms, veterinarians have to use tests performed at a diagnostic lab. The test that is currently used takes 14 days to complete. In this project, we will create a new diagnostic test that can be performed in less than a day. This new test will let the veterinarian know if the medication that they are using against the hookworms will work or not. This project will add to our knowledge about how much resistance we should expect to see in hookworms from dogs that live in the U.S. Knowing how much resistance to expect in a geographical area and knowing how many hookworms within a dog patient are specifically resistant will help veterinarians make important treatment decisions quickly.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 03169

RESEARCHERS

Jeba Jesudoss Chelladurai, BVSc, PhD
Kansas State University
Amount: $3,000

Using a heritable immunostimulatory bacterium to block heartworm transmission by mosquitoes

Summary: Researchers will investigate the use of a bacteria to help block transmission of heartworm parasites by mosquitoes.

Description: More than 150,000 cases of canine heartworm disease are reported annually in the United States alone. Monthly preventive treatments for dogs have been widely available since the mid-19805. However, drug-resistant strains are on the rise, highlighting the need for novel disease control strategies. Researchers will investigate how infection by certain bacteria decreases the capacity of mosquitoes to transmit infectious larvae of Dirofilaria immitis, the agent of heartworm disease. Bacteria-infected mosquitoes (and control groups) will be exposed to both drug-sensitive and drug-resistant parasites. The team hopes findings will provide a new approach to block the spread of the heartworm disease by targeting parasites in their mosquito host.

Co-sponsored with the Morris Animal Foundation, Grant Number: DZ2CA·015

RESEARCHERS

Michael Povelones; University of Pennsylvania
Amount: $10,000

Defining the Unique Genetic Markers in Dogs That Define Immune Function, Disease Resistance and Tissue Transplantation

The Major Histocompatibility Complex (MHC) genes encode proteins that are critical for a wide range of biological functions, from immune protection against infectious disease to the predisposition of an individual to develop diabetes and auto immune diseases.

The MHC genes in the dog are incompletely characterized, thereby severely limiting our ability to full define the cause of many canine diseases. Dr. Ramakrishnan has developed improved methods for identifying the different forms of canine MHC genes in a large number of dogs of diverse breeds.

In this study he will characterize the patterns of MHC genetic variation in over 1200 dogs from at least 50 breeds using a high throughput sequencing strategy. The distribution and frequency of different forms of each of these genes and their specific clustering among different breeds will greatly enhance our knowledge of the genetic diversity among breeds.

The methodology and data gained from this study will enhance the power of association studies between MHC types and canine diseases. Such a database will also enable tissue transplantation from unrelated but matched donors as a treatment for advanced malignancies (stem cell transplants) and other diseases (tissue transplantation). Fully defining the canine MHC will have broad impact across canine health, including oncology, immunology and infectious disease.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 01771

RESEARCHERS

Bevery Torok-Storb, PhD, DVM
Fred Hutchinson Cancer Research Center
Amount: $2,000

Developing a New Tool to Study Viral Infections and Cancer in Dogs

Development of an MHC Class I Tetramer to Study Virus- and Tumor-specific CD8+ T-cell Responses in Dogs

Summary: Researchers will develop a state-of-the-art molecular tool to track and study killer T-cell populations that are responsible for fighting viral infections and cancer in dogs.

Description: In humans, a powerful immunologic reagent called a tetramer is standardly used to visualize changes in the body’s killer T-cells. These cells respond to immunologic challenges and are critical to the body’s immune system.

Current knowledge of T-cell behavior in dogs could be significantly advanced with the development of a dog-specific tetramer. Researchers will work to construct the first canine tetramer, which would then be used in the development of vaccines for infectious diseases and cancer in dogs.

Co-sponsored with the Morris Animal Foundation, Grant Number: D15CA-015

RESEARCHERS

Dr. Paul R. Hess
North Carolina State University