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An Analysis of Canine Urine: Microbiota, Methods, and Changes in Health and Disease

Mrofchak, Ryan
http://rave.ohiolink.edu/etdc/view?acc_num=osu1620677605241405

Abstract Details

2021, Master of Science, Ohio State University, Comparative and Veterinary Medicine.
Urine was considered sterile up until the 21st-century; however, culture-independent methods and next-generation sequencing reveal urine to have a healthy and diverse microbial community. Within the past decade, compositional shifts in the urine microbiota have been observed in urinary diseases, such as bladder cancer. So far, studies on the urine microbiota have primarily focused on human urine. Dogs can serve as model organisms for urinary diseases like bladder cancer because the development and progression of the disease is similar to humans. Challenges arise in urine microbiota studies because urine contains low quantities of bacterial DNA. In objective one, host and bacterial DNA were extracted in urine samples from 10 healthy dogs using five extraction methods. Microbial diversity and composition were assessed through 16S sequencing. Of the five methods, the QIAamp BiOstic Bacteremia DNA Kit (Qiagen, Hilden, Germay) yielded the highest total (Kruskal-Wallis, p = 0.165, not significant) and bacterial (Kruskal-Wallis, p = 0.044) DNA concentrations. In addition, microbial diversity and composition differed by dog and sex, but not extraction method. The Bacteremia kit was then used to extract DNA in the urine of dogs with and without bladder cancer. In objective two, the microbial composition and diversity of urine samples from 7 dogs with urothelial carcinoma (UC) and 8 healthy dogs was assessed. Microbial composition differed between groups (PERMANOVA: Unweighted UniFrac, p = 0.032); however, differences in microbial diversity were not observed. There were no differentially abundant taxa between the two groups. These studies lay the foundation for future studies on the canine urine microbiota in healthy and sick dogs. Besides assessing microbes in urine, physical and chemical properties of urine are also routinely used to assess patient health. Understanding fluctuations of properties like pH or urine specific gravity in healthy urine are critical to determining when and how to assess urine from unhealthy individuals. Urine pH can be measured quickly and cheaply using dipsticks; however, this method is less exact than pH measured by pH meter. Urine specific gravity (USG) is an estimate of urine concentrations and can fluctuate throughout the day. In objective 3, urine pH and USG values were recorded in urine samples from five dogs over twelve timepoints ranging from hours to months apart. Urine pH varied widely over time within and between dogs while USG was more consistent by dog over time. pH as measured values via pH meter was significantly higher than pH as measured by dipstick (Wilcoxon Matched-Pair Rank Sign Test, p = 0.018), and these differences were clinically significant (>0.25) indicating that pH meters and dipsticks should not be used interchangeably. Unexpectedly, change). USG values did not differ between first morning and afternoon urine samples (Friedman’s test, p = 0.285). Collectively, our findings suggest the microbiota, urine pH, and USG vary within and between dogs, and that drivers of compositional change in the urine microbiota in both healthy dogs and dogs with bladder cancer warrant further investigation.
Vanessa Hale (Advisor)
Deborah Knapp (Committee Member)
Sheryl Justice (Committee Member)
159 p.
Animal Sciences; Microbiology
canine, urine microbiota, extraction methods, bladder cancer

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Citations

  • Mrofchak, R. (2021). An Analysis of Canine Urine: Microbiota, Methods, and Changes in Health and Disease [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1620677605241405

    APA Style (7th edition)

  • Mrofchak, Ryan. An Analysis of Canine Urine: Microbiota, Methods, and Changes in Health and Disease. 2021. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1620677605241405.

    MLA Style (8th edition)

  • Mrofchak, Ryan. "An Analysis of Canine Urine: Microbiota, Methods, and Changes in Health and Disease." Master's thesis, Ohio State University, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=osu1620677605241405

    Chicago Manual of Style (17th edition)

osu1620677605241405
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This open access ETD is published by The Ohio State University and OhioLINK.