Relationship between the level of reactive oxygen species in native rooster spermatozoa and the quality indicators of fresh and frozen-thawed sperm
Purpose: to study the influence of the level of active forms of oxygen in native spermatozoa of roosters on the qualitative indicators of freshly exposed and deconed individual ejaculators (mobility, damage to the cell membranes of sperm) and determining the permissible level of AFC generation to improve the cryoponement protocol.Materials and methods. The object of the study was the roosters of the Rodi-Aland Red (n = 20) breed at the age of 44 weeks of life. All males were kept in individual cells with the “Genetic Collection of Rare and Disappearing Course breeds” of VNIIGRZH systems adopted by the BRK for feeding, posting and light regime. Sperm was received in penicillin bottles with a volume of 10 ml, using the abdominal massage method. They measured the volume of each individual ejaculate, assessed the mobility of sperm, concentration. Cryoconservational was carried out in granules. Thawing of granules was carried out at T 60 ° C in a slit tie. The damage to the plasma membranes of sperm in the native and deconed seed was evaluated using the Suppitial Bluma coloring method. Spermatozoa with damaged membranes was painted red, intact cells remained white (colorless). Each drug estimates at least 200 cells. To determine the levels of AFC generation in spermatozoa of roosters, a method based on luminol-proroxidate hemilyuminescence was used, which was measured on a chemilyuminometer Lum-1200. The time of each measurement was 3 hours, based on the hemiluminiscence of the active form of oxygen (given the growth of the indicator, peak and decline). Cell concentration (7x106 classes/ml) was selected experimentally for measurements, according to the results of a series of preliminary experiments.Results. As a result of the study, data based on the method of luminol-proroxidate chemilyuminescence for the permissible level of AFC in native sperm of roosters were first obtained. The range of active forms of oxygen (from 75 to 249 volts*sec) has been established, in which cells do not receive significant damage to membrane structures during cryoponservation. In case of exceeding the threshold of 250 volts*S, the number of cells with damaged membranes increases sharply from 17.19% to 62.87%. The data obtained allow the assessment and selection of roosters on the quality of their sperm for the purposes of cryoponservation and the formation of cryobans of reproductive cells.