Influence of Technological Stages of Preparation of Rooster Semen for Short-Term and Long-Term Storage on Its Quality Characteristics

Abstract

There is a problem of declining quality of rooster semen in the “native semen-equilibrium-short-term and long-term storage (cryopreservation)” cycle. The aim of this study was to determine the effects of various methods of preparing rooster semen on its qualitative characteristics, taking into account the method of removing possible contaminants (centrifugation or filtration), and to evaluate the change in the composition of the cytosol of the spermatozoon of the native semen, during equilibration of the diluted semen and during short-term storage. In this study, semen from roosters (n = 22) of the Russian White breed was used. Experiment 1: semen was divided into 3 aliquots: I—was diluted with synthetic cryoprotective medium (1:1 with LCM control, II—was filtered (membrane pore Ø 0.2 μm), and III—was centrifugated (at 3000 rpm for 10 min). Native and frozen/thawed semen was evaluated. Experiment 2: the composition of carbohydrates and polyols of the spermatozoa of native semen was evaluated during equilibration and after storage (3 h). The results of Experiment 1 showed an advantage in the quality of filtered semen compared to centrifuged in terms of progressive motility (41.0% vs. 27.0%) and chromatin integrity (56.6% vs. 33.6%). Results from frozen/thawed samples of filtered semen compared to centrifuged in terms of progressive motility were 25.5% vs. 5.5%, respectively, and in terms of chromatin integrity—83.5% vs. 64.4%, respectively. The results of Experiment 2 showed the main component in the composition of the native spermatozoa cytosol in assessing the content of carbohydrates and polyols was inositol—75.6%. The content of inositol decreased during storage by 6.5 times (from 0.030 mg/mL to 0.007 mg/mL), proposing the role of inositol as the main antioxidant in the cytosol of spermatozoa, which makes it biologically justified to introduce inositol into the composition of synthetic diluents, including cryoprotective ones.

The Effects of Equilibration Time and Post-Thawing Temperatures in Cryopreservation of Gaga Chicken Semen

“…Then observe with a magnification of 400X-microscope in ten fields of view or 200 cells of sperm. The dead sperm will absorb red matter because the permeability of the cell wall has been weakened while the living sperm would look transparent (Silyukova et al, 2022).…”

“…The semen was placed on a glass of the object and then dried and scrubbed for 30 minutes in a 5% formalin solution at a temperature of 37 °C, rinsed with distilled water and air-dried. The dyeing procedure is based on Silyukova et al (2022) that semen was dried for 5 minutes in a CBB solution (0.25% Coomassie Brilliant Blue R 250 in a solution of 10% glacial acetate acid and 25% methanol) and then rinsed with distillation water and dried with air. Observations of acrosome integrity are done at 5-6 fields of view in a 100x magnification microscope.…”

“…CB stain was commonly used in protein identification. Coomassie Blue is able to bind to membrane proteins, by binding to glycoprotein (Silyukova et al, 2022). The spermatozoa acrosome membrane consists of the Inner Acrosome Membrane (IAM) and the Outer Acrosome Membrane (OAM) which serves as the protector of the sperm of acrosome that will be coloured (Khawar et al, 2019).…”

Supplementation of Glycine and Glucose into Egg Yolk Lactated Ringer Diluent on The Quality of Local Chicken Semen Stored at 5oC for 120 Hours

“…Subsequently, they were immersed in Coomassie brilliant blue -R 250 solution for 5 minutes, rinsed again with distilled water, and dried. The preparations were observed under a microscope at 1000× magnification, assessing a minimum of 10 fields of view (Silyukova et al, 2022).…”

The addition of egg yolk to the physiological saline extender improved the motility and viability of kampung rooster spermatozoa at cool temperatures

“…Selective mating between roosters and hens, resulted in a limited number of mated hens, thereby, the spermatozoa of roosters are not utilized optimally (Hidayat and Asmarasari, 2015). The artificial insemination technique allowed one male ejaculate to fertilize 24-40 females (Silyukova et al, 2022). By using superior roosters, artificial insemination technology can increase the population, productivity and genetic quality of kampung chickens.…”