Author ORCID Identifier

https://orcid.org/0000-0001-9130-4752

Date Available

7-24-2020

Year of Publication

2019

Degree Name

Doctor of Philosophy (PhD)

Document Type

Doctoral Dissertation

College

Agriculture, Food and Environment

Department/School/Program

Veterinary Science

First Advisor

Dr. Barry Ball

Abstract

Pregnancy loss remains a major source of economic cost to the equine industry. Frequently, the exact causes of pregnancy loss remain unknown. It has been shown, in other species, that increased dietary protein leading to elevated blood urea nitrogen concentrations (BUN) can be a factor in decreased survival of the early embryo. Our studies provided novel information regarding the effects of elevated BUN on endometrium and embryos from mares as well as insights on changes in their gene expression. Our first objective was to develop an experimental model to elevate BUN during diestrus using intravenous urea infusion. We analyzed the effects of an acute elevation in BUN on uterine and vaginal pH along with changes in the endometrial transcriptome of mares with RNA sequencing. There was a significant increase in BUN and a decrease in uterine pH in the urea group compared to the control group. A total of 193 genes were differentially expressed (DEG) between the urea and control groups. The DEG were predicted to be related to cell pH, ion homeostasis, changes in epithelial tissue, fatty acid metabolism, and solute carriers. Our second objective was to evaluate the effects of elevated BUN in the endometrium of mares using a chronic oral urea administration to elevate BUN in mares. Uterine and vaginal pH were evaluated and RNA sequencing of the endometrium was again performed. There was an increase in BUN in the urea-fed mares, but no significant change in uterine or vaginal pH between the groups. A total of 60 DEG were characterized, with prediction of transcriptomic changes in the endometrium of mares related to cell death (necrosis) and cellular movement (invasion of cells). Our third objective was to determine the effects of a high BUN on the transcriptome of day-14 embryos. There was a positive correlation between plasma BUN and blastocoele fluid urea nitrogen concentration. Changes in embryo transcriptome were related to survival of organism, angiogenesis, adhesion, and quantity of cells. Our final objective was to evaluate the correlation between BUN and follicular fluid urea nitrogen and evaluate the survival of embryos collected from donor mares with high BUN concentrations. Urea nitrogen concentration was positively correlated between the plasma and follicular fluid of mares. Additionally, there was a higher pregnancy rate when embryos were collected from mares with lower BUN. Overall, these results further elucidate the mechanisms through which urea affects endometrial and embryonic transcriptome of mares with high BUN, serving to identify effects of a high BUN in the reproductive tract of mares that might lead to decreased fertility.

Digital Object Identifier (DOI)

https://doi.org/10.13023/etd.2019.301

Funding Information

This work was funded by Science without Borders- CNPq- GDE- USA, Grant/Award number: 208518/2014-2 and USDA-ARS National Program 215, Grass, Forage and Rangelands Agroecosystems (project number 5042-21000-003-00D).

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