Analysis of vitality and biochemical parameters in freeze-thawed seminal plasma of rams

Abstract

The current study aimed to examine the percentage changes of viability sperm and the activity of the enzymes LDH, ALP, GOT / AST, GPT / ALT in the sperm plasma of Lacaune rams, before and after cryopreservation. For this purpose, five rams were examined, and two ejaculates were obtained from each ram. Ejaculates are collected by the method of artificial vagina, during the insemination campaign. All ejaculates were diluted with a 6AG extender and frozen by the Cassou’s sequin method. Sperm viability was determined by eosin and nigrosine smears, and enzyme activity was examined spectrophotometrically. As a result, the percentage of vital sperm after cryopreservation decreased by 15% (P ≤ 0.001). The freezing and thawing process also reduced the activity of the enzymes LDH, ALP, GOT / AST and GPT / ALP. In conclusion, the observed enzymes, in relation to sperm vitality, could be used as indicators to optimize the protocols for cryopreservation of ram’s sperm.

Istraživanje je imalo za cilj ispitivanje promena u procentu vijabilnih spermatozoida i promena u aktivnosti enzima LDH, ALP, GOT / AST, GPT / ALT u seminalnoj plazmi ovnova rase Lakon, pre i nakon krioprezervacije. Za tu svrhu, ispitivana je sperma pet ovnova, sa dva ejakulata po ovnu. Ejakulati su dobijeni metodom veštačke vagine, u toku sezone parenja. Svi ejakulati su razređeni razređivačem 6AG i zamrznuti po metodi Cassou sequin. Vijabilnost spermatozoida je određena na osnovu razmaza sa eozinom i nigrozinom, a aktivnost enzima je određena spektrofotometrijski. Rezultati istraživanja pokazuju da je udeo vitalnih spermatozoida nakon krioprezervacije smanjen za 15% (P ≤ 0.001). Procesi zamrzavanja i odmrzavanja takođe smanjuju i aktivnost enzima LDH, ALP, GOT/ AST i GPT / ALP. Ispitivani enzimi, združeno sa vitalnošću spermatozoida, mogu da se koriste kao markeri za optimizaciju stope hlađenja i koraka u procesima zamrzavanja i odmrzavanja sperme ovnova.