Canine Semen Microbiome

Insufficient male reproductive performance and subfertility are important problems for dog breeders, but the causes underlying acquired infertility cannot be identified in about 50% of dogs. In humans, recent studies have demonstrated an association between semen quality and bacterial communities in the ejaculate, but further research is needed to assess a causal relationship between semen microbiome and semen quality.

Dog semen is not sterile and contains up to 10^5 bacteria/mL. Although previous studies showed no influence of some bacterial species such as Escherichia coli, Staphylococcus aureus, Klebsiella spp., Mycoplasma canis, Pseudomonas aeruginosa, and Streptococcus spp. on conception rates in dogs, it has been suggested that a higher overall concentration of bacteria and the presence of potentially pathogenic microorganisms might be associated with lower semen quality. Furthermore, antimicrobials are often misused in canine reproduction to address infertility problems. Previous research on dog semen microbiome is limited and relied only on culture, potentially missing some pathogens. Next generation sequencing (NGS) allows to describe novel microbiomes without prior knowledge of sequencing information or specific culture requirements. These techniques have been recently used to investigate the seminal microbiome of humans and domestic animals.

The research team’s aim is to perform a comprehensive investigation on the seminal microbiome of male dogs, as such identifying possible links between the seminal bacterial population and different semen parameters, opening a new chapter in canine andrology.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 03165-A


Penelope Banchi, DVM; Ghent University
Amount: $2,000

Evaluating Reproductive Diseases in vitro with a 3D Canine Endometrial Organoid Model

Pyometra is a life-threatening disease that affects over 66% of intact older bitches. Bitches are pre-disposed to this disease due to a number of unique features of their reproductive cycle. Endometritis is an underdiagnosed condition in bitches that results in reduced fertility and can lead to the development of pyometra.

These diseases have mostly been studied in live dogs resulting in welfare concerns and limitations in our understanding of the disease onset and progression and importantly, prevention of these conditions. Organoids are miniature organs in a culture dish that can be grown long-term while maintaining the characteristics and function of the original organ.

This unique 3-dimensional (3D) structure facilitates the study of disease processes such as endometritis and pyometra in canines, potential treatments, and assessment of preventative therapeutics such as novel vaccines. Optimization of canine uterine organoids has the potential to: a) improve the health and welfare of intact middle-aged and older female dogs; b) enhance fertility, genetic gain and health in working dog programs; and c) allow the study of female reproductive diseases without the use of research dogs.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 03055


Fiona Hollinshead, BVSc, PhD; Colorado State University
Amount: $1,000

Artificial Intelligence and Machine Learning for Diagnosis of Dog Sperm Morphology

Sperm morphology evaluation is an important component of dog fertility analysis. However, visual evaluation of sperm morphology is difficult to teach, and results are largely dependent on the proficiency and experience of the evaluator, leading to large variability in results within and across evaluators.

The objective of this pilot study is to explore recent advances in artificial intelligence for image pattern recognition, similar to that used by the likes of Google and Facebook, to create an automated method for sperm morphology evaluation. A large database with 10,000 dog sperm images will be created. The images will be evaluated by expert veterinarians, then used to train a Convoluted Neural Network (CNN) using deep learning methods. Investigators expect to generate an algorithm capable of classifying sperm morphology with greater than 90% accuracy and precision.

Employing such an algorithm could allow veterinarians to establish more reliable reference values and guidelines for prospective stud dogs and for semen to be used for artificial insemination, improve veterinarian’s abilities to formulate diagnoses and prognoses for infertility problems related to specific sperm defects, and to establish the reproductive safety of drugs and compounds.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 03045-A


Leonardo Brito, DVM, PhD; University of Pennsylvania
Amount: $1,000

Evaluation of a New Vaccine for Canine Brucellosis

Canine infection by Brucella spp. constitutes a serious problem for dog breeders and pet owners, leading to the economic burden associated with reproductive loss and veterinary care. Canine brucellosis is also considered a public health concern because of its potential to be transmitted to humans.

Within the US, the disease has reemerged due to the chronic persistence of the organism, low dose for infection, low sensitivity and specificity of the current diagnostic tests, and most importantly, the lack of a protective vaccine for dogs. Historically in the US, brucellosis control efforts for cattle, sheep, goats and domestic pigs have been successful mainly due to the availability of protective and efficacious vaccines.

The goal of the proposed research is to develop a brucellosis vaccine that is safe, stable, free of side effects and efficacious for dogs. Previous CHF funding (Grant #2275-A) has permitted the investigators to successfully engineer a promising live attenuated vaccine candidate, denominated B. canis RM666ΔvjbR.

This study will further investigate the ability of the vaccine candidate to induce appropriate immunity and will also develop a diagnostic assay capable of differentiating naturally infected vs vaccinated animals, necessary for mass vaccination. The development of a safe and highly protective brucellosis vaccine for dogs will significantly impact owners, breeders and human health by limiting the spread of the disease.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02441


Angela Arenas, DVM
Texas A&M AgriLife Research
Amount: $3,000

Genome sequencing and antimicrobial susceptibilities of Escherichia coli isolated from clinical cases of canine pyometra

Pyometra is a potentially life-threatening infection of the canine uterus by bacteria, most commonly Escherichia coli (E. coli). In humans with recurrent infections, E. coli produces biofilm, a layer of polysaccharide that protects the organism from the host immune system as well as antibiotic agents, decreasing treatment efficacy. Current treatments for pyometra are costly, time-consuming, and not without risk to the bitch.

The investigators postulate that biofilm production by E.coli within the uterine lining may be responsible for perpetuating the disease and making treatment difficult. In previous CHF-funded study, the investigators were able to prove that E. coli from clinical cases of canine pyometra is capable of producing biofilm both in the uterus and in laboratory settings.

The purpose of this study is to characterize the presence of ten different genes associated with biofilm production and disease-contributing factors of E. coli organisms to determine if there is an association with those strains of E. coli that produce biofilm and certain disease factors found in other strains of E. coli. Disease factor genes and resistance patterns will be identified, and may serve as targets for new therapeutics directed at the disruption of biofilm in an effort to shorten the duration of treatment of pyometra.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02512-A


Erin E Runcan, DVM
Ohio State University
Amount: $2,000