The improvement of preservation methods of rooster genome in a deep-frozen state is of great economic importance, since the artificial semination by means of cryopreserved semen practiced in poultry farming allows to expand the possibilities for creation of new breeds and hen lines, and to obtain a maximum amount of progeny from highly valuables in the breeding respect sires. Hydrophoby, the affinity of substance with water, is responsible for the molecular interaction and physico-chemical processes during the preservation of the biological material. In the experiments the rooster semen of Rhode-island breed was used. The cryopreservation of the semen was done using the granulation method. The hydrophobic interaction state was determined by the number of hydrophobic aminoacids. The semen was thawed out, using an aerodynamic installation. The quality of the cryopreserved semen was evaluated by the mobility of gametes expressed in 10- point system. The results of hen semination were evaluated by the total amount of incubated eggs. The goal of the study was to identify of hydrophobic interaction changes in the process of cryopreservation of the rooster semen and to raise the preservation efficiency by creating new cryoprotective media. The obtained results show that there is a direct dependence between the hydrophobic interactions and the mobility of the preserved gametes. The revealed changes of the hydrophobic interaction in the process of cryopreservation point to the fact that low temperatures can act as a factor, causing changes in the investigated parameter. These changes are of diversified nature and depend upon the study material. It can be explained by different structure of protein complexes due to intermolecular interactions in biological system and the disturbance of barrier functions of plasmatic membranes. Also, the performed studies showed that the use of isotonic solutions of nonelectrolytes for the preservation of rooster semen made it possible to achieve high indicators of gamete mobility after thawing out. The highest indicators were obtained in cases when the cryoprotective media contained maltose disaccharide. When the cryoprotective medium contains lactose, the quality of the preserved material becomes worse. These data allow ascertaining that the quality of the thawed out rooster semen depends not only on the quantity of glycoside radicals in a carbohydrate molecule but also upon its chemical nature. It was also shown that it is feasible to increase the mobility of the preserved semen by introducing arginine into the cryoprotective medium. On the basis of the experimental studies a new type of cryoprotective medium was developed which contained maltase 11,5g, glycerin 7ml, bidistilled water up to 100ml, arginine up to pH 6,8–6,9. A comparative experiment was carried out to evaluate the quality of the preserved semen, using the optimal medium The obtained data show that the mobility of the thawed out gametes after their preservation in the experimental medium reached 6,8±0,14 points which by 11,4% higher than the mobility of gametes in the check experiment. Based on the obtained under laboratory conditions results of the recovery of gametes mobility and the preservation of their acrosomal structures, an experiment was carried out under production conditions. It revealed that the use of maltase- arginine-glycerin medium for the cryopreservation of rooster semen makes possible the stabilization of its functional indicators, reaching an egg fertilization up to 70,8±6,56%. In the control samples the studied parameter was by 16,2% lower. Thus, hydrophobic interactions can be considered as a significant factor predetermining the gamete cryogenic resistances; improving of the cryoprotective medium carbohydrate contain allows increasing the efficiency of rooster sperm conservation.