Performing Department
Biomedical & Diagnostic Sciences
Non Technical Summary
Tritrichomonas foetus is a multi-flagellated parasite typically found in the urogenital tract of cattle but recently has emerged as an important pathogen in cats. T. foetus infection in cats produces prolonged and intractable diarrhea, often leading to large bowel disease. The incidence of T. foetus infection in cats has skyrocketed in recent years, and typical anti-protozoal treatments have been ineffective. Further complicating the study of T. foetus in cats, a variety of studies have found that genetic analysis depends highly on the gene chosen for analysis. Therefore, it is important to systematically characterize the differences in T. foetus strains infecting felines and bovines to develop more effective therapeutics or preventative strategies. Because T. foetus in cats has been shown to cause disease in cattle; however, bovine T. foetus has not been shown to cause disease in felines,we hypothesize that the feline and bovine isolates are distinct species. We further hypothesize that genetic differences in physiologically relevant genes may explain the observed differences in anti-protozoal activity and host preference between the isolates. Additionally, we speculate that rodents are the most likely route of infection for feline trichomoniasis, based on detection of a novel intestinal tritrichomonad isolated from rats. To test these hypotheses, we will analyze the whole genomes of two T. foetus strains, one bovine and one feline. Whole genome sequencing will allow verification that these are distinct species and also allow identification of areas of interest within the genome that can be used for targeted sequencing to compare numerous bovine and feline T. foetus isolates. The final objective will be to compare the genetic relatedness of the bovine and feline T. foetus with the rat tritrichomonad, to test the hypothesis that the feline isolates are more closely related to a tritrichomonad isolated from rats than T. foetus from cattle. Associated with all objectives will be the training of a PhD student, included in the proposal budget, which will meet the MAF goal of producing future veterinary/animal health researchers. Through the fundamental knowledge obtained through this study, it will be possible to better understand feline trichomoniasis, leading to the selection of better therapeutics, and overall increasing the quality of life for infected felines. Morris Animal Foundation; 01/01/2014 - 12/31/2017; $105,360
Animal Health Component
50%
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Goals / Objectives
Analyze the whole genomes of two T. foetus strains, one bovine and one feline. Verificationif these are distinct species andidentify areas of interest within the genome that can be used for targeted sequencing to compare numerous bovine and feline T. foetus isolates. Compare the genetic relatedness of the bovine and feline T. foetus with the rat tritrichomonad, to test the hypothesis that the feline isolates are more closely related to a tritrichomonad isolated from rats than T. foetus from cattle.Traina PhD student.
Project Methods
Using the single cell clones established in the pilot study,we willsequence the whole genome of one feline and one bovine isolate of T. foetus. Upon completion of the sequencing run, the genomes will be assembled. To aid in assembly, an optical map for a bovine T. foetus reference strain will be conducted in collaboration with OpGen. Optical mapping is used to create a map of restriction enzymes sites along unknown DNA, and has been used extensively to aid in the assembly of microbial and eukaryotic genomes. Upon assembly of the genomes, comparisons will be made between the bovine and feline isolates. After identifying genetic differences between the genomes, Sanger sequencing of target genes will be used to examine the differences between multiple feline and bovine isolates, and also a novel rat T. foetus. Finally, the genomes will be evaluated to determine if there are any consistent genetic differences that may correlate with drug resistance, with the ultimate goal of developing molecular therapies to treat these resistant infections.