Effect of semen washing on thawed ram spermatozoa subjected to a four hour post-thawing thermal evaluation test

Autor: Uchebuchi Ike Osuagwuh, M.J. Palomo
Rok vydání: 2017
Předmět:
Zdroj: Small Ruminant Research. 155:81-86
ISSN: 0921-4488
DOI: 10.1016/j.smallrumres.2017.09.010
Popis: This study was conducted to evaluate sperm motility and qualitative characteristics of ram spermatozoa cryopreserved in the presence or absence of seminal plasma under controlled condition and subjected to a 4 h post-thawing thermal evaluation test. The aim being that exposing these frozen-thawed sperm to thermal resistance test will provide useful information on their survivability in the female genital tract towards fertilizing the ovum. Briefly, ejaculates from 5 males (5years) were collected by artificial vagina, and split into two aliquots. One aliquot was diluted (1:5) in Tris-citric acid-glucose (TCG) solution and washed twice by centrifugation at 600× g for 10 min, while the other aliquot was kept unwashed. Thereafter, washed and unwashed sperm were extended in a TCG-based media containing 5% glycerol and 15% powdered egg yolk (PEY) and frozen. Frozen-thawed sperm were thawed and incubated at 37 °C for 4 h. Sperm motility characteristics was evaluated by CASA while plasma membrane integrity (SYBR-14/PI), acrosome integrity (PE-PNA), mitochondria activity (Mitotracker deep red) and reactive oxygen species (H 2 DCFDA) were analysed by flow cytometry. Sperm nucleoprotein integrity assessment was done by the determination of overall levels of sperm-head disulfide bonds via spectrophotometric analysis of free-cysteine radicals’ levels. Unwashed samples generally showed better results than washed samples as regards to sperm motility characteristics irrespective of incubation. The post-thawing incubation had a significant (P
Databáze: OpenAIRE
Externí odkaz: