title: Effect of dilution rate on feline urethral sperm motility, viability, and DNA integrity.
abstract: This study was designed to investigate if the characteristics of feline urethral sperm can be affected by high dilution in an artificial medium. The semen collected by urethral catheterization from eight male cats was evaluated for sperm concentration and motility and subsequently diluted with a TRIS-based extender to the concentration of spermatozoa 10 × 10(6)/mL, 5 × 10(6)/mL, and 1 × 10(6)/mL. Immediately after the extension samples were assessed for motility, cell viability using SYBR-14 and propidium iodide, acrosome integrity using lectin from Arachis hypogaea Alexa Fluor 488 Conjugate, and propidium iodide and chromatin status by acridine orange. Compared with 10 × 10(6)/mL dilution rate, spermatozoa diluted to 1 × 10(6) sperm/mL had a significantly lower proportion of motile (31.1% ± 19.8 and 0.7% ± 1.6, respectively, P < 0.05) and viable spermatozoa (88.3% ± 3.1 and 69.1% ± 12.8, respectively, P < 0.01). There was no dilution-related difference in the acrosome integrity (76.7% ± 11.9 vs. 75.9% ± 10.6) and chromatin status (defragmentation index, 3.3% ± 0.97 vs. 3.4% ± 1.7). These results indicate that feline urethral semen is susceptible to high dilution rate, and some sperm characteristics can be artifactually changed by semen dilution. It also suggests the potential role of seminal plasma in maintaining sperm motility and viability in high dilution rates.
authors.affiliation: Department of Reproduction and Clinic of Farm Animals, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Prochowska
authors.affiliation: Department of Reproduction and Clinic of Farm Animals, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland. Electronic address: wojciech.nizanski@up.wroc.pl.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Niżański
authors.affiliation: Department of Reproduction and Clinic of Farm Animals, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Ochota
authors.affiliation: Department of Reproduction and Clinic of Farm Animals, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Partyka
mesh_terms: D000818:Animals; D002415:Cats / Q000235:genetics / Q000502:physiology*; D004249:DNA Damage; D005434:Flow Cytometry; D008297:Male; D013048:Specimen Handling / Q000379:methods / Q000662:veterinary; D013081:Sperm Motility; D014521:Urethra / Q000166:cytology
keywords: Cat semen; Dilution effect; Flow cytometry; Sperm concentration