Identifier

etd-07132015-125201

Degree

Master of Science (MS)

Department

Biomedical and Veterinary Medical Sciences - Veterinary Clinical Sciences

Document Type

Thesis

Abstract

The objectives of this study were to investigate concentration dependent effects of canine prostatic fluid (PF) on in vitro seminal parameters of cooled canine semen. Sperm motility parameters, plasma membrane integrity and stability, acrosome integrity and DNA fragmentation were measured after the addition of 0%, 10%, 25%, or 50% PF to extended semen of fertile dogs. Assessments were made at 0 h pre-cooling, at 24, and 48 h of cooled storage (4 °C), and after freezing and thawing followed by incubation (37 °C) at 0, 4 and 24 h. Our hypothesis was that lower dilutions of canine semen with PF in an egg yolk-Tris extender would improve plasma membrane stability and acrosome integrity, and preserve sperm kinetics and reduce DNA fragmentation in comparison with higher concentrations of PF in fertile dogs during cooling. Sperm motility parameters were assessed by computer assisted sperm analysis, and plasma membrane integrity by the hypo-osmotic swelling test. Flow cytometry was used after staining with YO-PRO-1/Ethidium Homodimer 1 (EthD-1) to evaluate membrane stability, fluorescent isothiocynate-PNA (Arachis Hypogaea)/propidium iodide to assess acrosome integrity, and sperm chromatin structure assay to assess DNA fragmentation. The data was analyzed using a mixed linear model (ANOVA) and in case of significant effects of time, treatment, or treatment*time interaction (P < 0.05), least square means were used for pairwise comparisons. Acrosome integrity and DNA fragmentation were not affected by treatment with PF. During the cooling period motility parameters were not influenced by PF treatment. A lower proportion of early apoptotic and higher proportion of early necrotic cells was seen during cooling with 50% PF (YO-PRO-1/EthD-1). Although lower concentrations of PF did not improve the evaluated spermatozoal parameters, they did not seem to compromise sperm motility and plasma membrane stability. The presence of 50% PF prior to cryopreservation decreased post thaw motility and produced a shift towards early necrotic cells after thawing. Therefore admixture with more than 10% PF should be avoided prior to cryopreservation of canine semen.

Date

2015

Document Availability at the Time of Submission

Student has submitted appropriate documentation to restrict access to LSU for 365 days after which the document will be released for worldwide access.

Committee Chair

Lyle, Sara

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