Cryopreservation of ram-sires semen and its qualitative changes under the influence of low temperatures


Keywords: breeding rams, semen, cryopreservation, thawing, equilibration, cryoprotectants, fertilizing ability.

Abstract

The article presents in retrospect the methods of cryopreservation of sperm of bulls and breeding rams. It is indicated that the first methods were proposed by I.V. Smirnov in 1947–1951, which ensured the production of viable lambs and calves. The significant contribution to the development of the method of cryopreservation of bull sperm by British scientists Polge C. and Rawson L. and Japanese scientists Nagase H. and Niwa T. The methods developed by them, with minor changes, have been used by scientists and breeding specialists for many decades. Modern technologies of cryopreservation of bull sperm are represented, in particular: Kharkiv in coated granules (author FI Ostashko and others), French company IMV-R-Cassou, German company Minitub, which are currently used in most countries with developed animal husbandry. It is noted that the first attempts to freeze the sperm of breeding rams were made by E. M. Platov in 1965. It is noted that in the cryopreservation of sperm the importance of adaptation and equilibration processes is important. The role of glycerin as a cryoprotectant in freezing the semen of male farm animals has been elucidated. The optimal concentration of glycerin in the media for cryopreservation of sperm of fetuses has been established. It is noted that the basis of “temperature shock” of sperm is damage to the sperm membrane, which is caused by balancing the concentration gradient that occurs at the boundary of cell-medium division with a sharp change in temperature. A high correlation between the physiological quality of the acrosome of sperm and its fertilizing ability is indicated. Studies of the chemical composition of the sperm of breeding rams have shown that it differs from the sperm of other animal species by a slightly higher content of proteins, lipids of lactic acid and a lower concentration of minerals. Proteins contain much more amino acids. Sheep sperm do not tolerate aerobic conditions, it is proved that damage to sperm by oxygen occurs mainly after its dilution and equilibration, and not during cryopreservation. Under aerobic conditions, one amino acid is cleaved from the molecule of lecithin, which is part of the membranes of sperm, with the formation of toxic lysolecithin. The most critical zones at thawing of deep-frozen sperm are shown. It has been experimentally established that the rate of the recrystallization process can be reduced by using cryoprotectants in environments, as well as by increasing the rate of thawing. An important influence in the thawing of sperm has the selection of the optimal mode of thawing. Analysis of literature data revealed that in practical conditions in the process of freezing ‒ thawing about 50% of sperm lose activity, which indicates that currently available methods of freezing sperm of breeding rams are imperfect and need further study and improvement.

References

Aybazova, M. M. (1987). Usovershenstvovanie tehnologii kriokonservatsii spermyi i ispolzovanie tsennyih baranov-proizvoditeley. Avtoreferat diss. kand. vet. nauk. Stavropol. SSHI (in Russian).

Aybazov, M. M. (2004). Biotehnologiya vosproizvodstva ovets i koz: monografiya (in Russian).

Belous, A. M., Bondarenko, V. A., & Shrago, M. G. (1982). Strukturnyie izmeneniya biologicheskih membran pri ohlazhdenii. Nauk. dumka (in Russian).

Berndtson, W. E., & Foote, R. H. (1972). The freezability of spermatozoa after minimal pre-freezing exposure to glycerol or lactose. Cryobiology, 9(1), 57–60. doi: 10.1016/0011-2240(72)90010-7.

Blom, E. (1964). The galea capitis as pert of the acrosome in the bull sperm. Animal Reprod. Trento, 4, 655–666.

Blom, E. (1964). Spermatic ultrastructure (bovine). Spermatic ultrastructure (bovine). 3. Edition springfield, Nass, 425–430.

Bugrov, A. D., & Olekseenko, T. N. (1977). Kriticheskie zonyi ottaivaniya spermyi byikov i ih vliyanie na vyizhivaemost spermiev. Selskohozyaystvennaya biologiya, 13(3), 368–370 (in Russian).

Buhrov, O. D. (1965). Vplyv osmatychnoho tysku na simia plidnykiv pry yoho hlybokomu zamorozhuvanni. Zb. Rozvedennia i utrymannia silskohospodarskykh tvaryn, 5, 36–41 (in Ukrainian).

Cassou, R. (1964). La méthode des paillettes en plastique adaptée a la généralisation de la congélation. In Fifth International Congress on Animal Reproduction and Artificial Insemination (Trento), 4, 540–546.

Danov, T. D., Kovachev, K. D., & Kresteva, S. H. (1984). Nakoi formi na uvrezhdane i otklonenie ot normalnata Ultrastruktura na spermatozoidite na kocha. Biol. Et immunal. Reprod, 9, 87–94.

Davidenko, V. M., Shinkarenko, I. S., & Ignatenko, O. I. (1977). Vliyaniya nekotoryih faktorov na kachestvo spermyi pri zamorazhivanie. Nauchno-tehn. Byull. Ukr. NIIZh shtatnyih rayonov “Akanya Nova”, 41, 60–63 (in Russian).

Emmens, C. W., & Blackshow, A. W. (1955). The fertility of frozen ram and bull semen. Austral. Veter. I, 31(3), 76–79. doi: 10.1111/j.1751-0813.1955.tb05502.x.

Fornusek, L., Vetvickova, V., & Petelikova, I. (1981). Usinek redidel pro dlouhodobe uchovavani na akrosomy beranich spermii. Veter.-Med. (CSSR), 26, 213–221.

Habibullin, H. H. (1957). Sohranenie i perevozka spermyi byikov pri nizkih temperaturah. Molochnoe i myasnoe zhivotnovodstvo, 5, 39–42 (in Russian).

Habibullin, H. H. (1958). Hranenie semeni byika v zamorozhennom sostoyanii. Molochnoe yi myasnoe zhivotnovodstvo, 2, 43–47 (in Russian).

Hammarstedt, R. H., Amman, R. P., & Ruinsky, T. (1976). Biol. Reproductions, 14, 381–387.

Harrison, R. A. P., & Flechon, J. E. (1980). Immunocytochemical detection of acrosomal damage following cold shock: loss of acrosin from the acrosomal region of ram, bull and boar spermatozoa. Reproduction Nutrition Développement, 20(6), 1801–1810. doi: 10.1051/rnd:19801006.

Ilinskaya, T. P. (1963). O rezultatah zamorazhivaniya semeni byikov v Belorussii. V kn. Novoe v plemennom dele v iskusstvennom osemenenii selskohozyaystvennyih zhivotnyih, 389–394 (in Russian).

Jodent, R. (1976). Recent technological progress in artificial insemination of cows. Italian. Selezione Veterinaria.

Jones, R. W. (1965). The use dimethyl sulfoxide glycerol and reconstituted skim milk for preservation of ram spermatozoa. Austral. J. Biol.Sci., 18(4), 877–885. doi: 10.1071/BI9650877.

Kalev, G., Zagorskiy, D., Bapirdzhiev, K. i dr (1970). Osemenenie ovets glubokozamorozhennoy spermoy, razbavlennoy razlichnyimi sredami. Zhivotnovodstvo, 7, 28–31 (in Russian).

Kantsedal, V. I. (1968). Nekotoryie voprosyi fiziologii i biohimii spermyi hryakov i baranov. Avt. dis.. kand. biol. nauk. Kazan (in Russian).

Kononov, V. P. (1983). Dostizheniya i perspektivyi zamorazhivaniya semeni v svinovodstve. M.: Rosselhozizdat (in Russian).

Korban, N. V., Moroz, L. G., & Shapiev, I. Sh. (1980). Metodicheskie rekomendatsii po nizkotemperaturnoy konservatsii spermyi hryakov (in Russian).

Korotkov, A. I., Marchenko, M., & Moskalyuk, M. (1960). Hranenie semeni baranov pri temperature -78° C. Zemledelie i zhivotnovodstvo Moldavii, 1, 42–44 (in Russian).

Kurilo, Yu. G. (1982). Vzaimosvyaz aktivnosti fermentov i kontsentratsii metabolitov uglevodnogo obmena v sperme hryakov s kachestvom spermyi, perezhivaemostyu i oplodotvoryayuschey sposobnosti spermiev (Doctoral dissertation, stupenya dok. biol. nauk: spets. 03.0013 “fiziologiya”. Harkiv (in Russian).

Kushnir, V. M. (1974). Deystvie osmoticheskih faktorov pri zamorazhivanii spermyi byikov v sredah s glitserinom Avtoreferat kand. diss (in Russian).

Kushnir, V. M. (1998). Perspektivyi razvitiya teorii i praktiki kriokonservatsii spermyi. Konkurento-sposobnoe proizvodstvo produktsii zhivotnovodstva v Respublike Belarus: Sb. rabot Mezhd.nauch, 146–148 (in Russian).

Kuznetsov, M. P., & Kupriyanova, L. P. (1959). K voprosu sohraneniya semeni baranov pri temperature -79° C. Tr. Vizha, 23, 171–190 (in Russian).

Kuznetsov, L. N. (1990). Opyit glubokogo ohlazhdeniya i dlitelnogo hraneniya semeni baranov. Novoe v plemennom dele i iskusstvennom osemenenii selskohozyaystvennyih zhivotnyih (in Russian).

Lopyirin, A. I., Loginova N. V. (1958). Metodike zamorazhivaniya semeni barana. Ovtsevodstvo, 8, 31–34 (in Russian).

Luyet, B. J. (1937). The vitrification of organic colloids and of protoplasme. Biodynamica.

Luyet, B. J., & Hodapp, E. L. (1938). Revival of frog's spermatozoa vitrified in liquid air. Proceedings of the Society for Experimental Biology and Medicine, 39(3), 433–434. doi: 10.3181/00379727-39-10229P.

Maksimov, N. A. (1913). O vyimerzanii vodyi i holodostoykosti rasteniy (in Russian)

Maksimov, N. A. (1929). Vnutrennie faktoryi ustoychivosti rasteniy k morozu i zasuhe. Trudyi po prikladnoy botanike, genetike i selektsii, 22(1), 3–41 (in Russian).

Mann, T. (1964). Biochemistry of semen of the male reproduct. Tract. Science, 147, 3659, 727–728. doi: 10.1126/science.147.3659.727-a.

Marinov, P. (1981). La accion proteoritika del acrosome como metodo de valoracion del efecto crioprotector de dillnyentes para la congelacion de semen ovino. Biol. Immunal. Reproduct, 5, 44–48.

Martin, J. C. A. (1963). The influence of diluente and processin time ofter ejaculation on the revival of deep frozen bull spermatozoa. J. Agric. Sei, 63, 55–63.

Martin, I. C. A. (1963). Effects of lecithin, egg-yolk, fructose and period of storage at 5° c on bull spermatozoa deep-frozen to-79° C. Reproduction, 6(3), 441–449. doi: 10.1530/jrf.0.0060441.

Matskepladze, I. B., Gugushvili, K. F., Bregadze, M. A i dr (1960). Sohranenie i ispolzovanie semeni byika i barana v zamorozhennom sostoyanii. Zhivotnovodstvo, 2, 77–79 (in Russian).

Mazur, P. (1966). Theoretical and experimental effects of cooling and warming velocity on the survival of frozen and thawed cells. Cryobiology, 2(4), 181–192. doi: 10.1016/S0011-2240(66)80165-7.

Mesanos, P., Wamsik, R., & Schavac, F. (1977). Prezivatel spermii barana pri pouziti roznych bruhov riedidiel pohlbokom zmrazeni v tekutom dusuku. Vet. Med, 22, 599–604 (in Russian).

Miller, W. J., & VanDemark, N. L. (1954). The influence of glycerol level, various temperature aspects and certain other factors on the survival of bull spermatozoa at sub-zero temperatures. Journal of Dairy Science, 37(1), 45–51. doi: 10.3168/jds.S0022-0302(54)91230-8.

Milovanov, V. K. (1962). Biologiya vosproizvedeniya i iskusstvennoe osemenenie zhivotnyih. Kolos (in Russian).

Milovanov V. K. (1975). Eksperiment k bionike epididimisa (k tehnologii hraneniya semeni selskohozyaystvennyih zhivotnyih). Dokladyi VASHNIL, 10, 26–28 (in Russian).

Milovanov, V. K., & Sokolovskaya, I. I. (1980). Dlitelnoe hranenie semeni barana i novyie vozmozhnosti krupnomasshtabnoy selektsii v ovtsevodstve. Vestnik selskohozyaystvennoy nauki, 2, 122–131 (in Russian).

Milovanov, V. K., & Sokolovskaya, I. I. (1980). Uroven i perspektivyi nauchnyih issledovaniy. Zhivotnovodstvo, 10, 48–50 (in Russian).

Milovanov, V. K., Varnavskiy, A. N., & Nauk, V. A. (1970). O prirode kriogennyih povrezhdeniy zhivchikov barana. Vestnik s-h nauki, 10, 86 (in Russian).

Milovanov, V. K., Varnavskiy, A. N., & Varnavskaya, V. A. (1976). Razrabotka metoda glubokogo zamorazhivaniya semeni baranov dlya dlitelnogo hraneniya. Zhivotnovodstvo, 8, 57 (in Russian).

Mollova, M. V. (1980). Changes in the structure of ram spermatozoa after equilibration and deep freezing. In Doklady Bolgarskoi Akademii Nauk, 33(3), 393–395.

Moroz, L. G., Shapiev, I. Sh., Korban, N. V., & Mashanskiy, V. F. (1976). Fiziologicheskie i biohimicheskie izmeneniya spermatozoidov hryaka pri zamorazhivanii. Selskohozyaystvennaya biologiya, 11(6), 850–856 (in Russian).

Morozov, V. A. (1957). Sohranenie semeni barana v zamorozhennom sostoyanii posredstvom gipertonicheskih rastvorov. Dokladyi VASHNII, 11, 44–48 (in Russian).

Mykytyn, L. Ye., Binkevych, V. Ya., & Bilyk, O. Ya. (2013). Stan ta perspektyvy rozvytku vivcharstva v Ukraini. Naukovyi visnyk Lvivskoho natsionalnoho universytetu veterynarnoi medytsyny ta biotekhnolohii im. Gzhytskoho, 15, 1(4), 133–141 (in Ukrainian).

Nagase, H. (1964). Deep-freezing bull semen in concentrated pellet form. I. Factors affecting survival of spermatozoa. In Proceedings of the 4th International Congress on Animal Reproduction and Artificial Insemination, 3, 410–415.

Nauk, V. A. (1973). Sohraneniya semeni barana i izmenenie soderzhanie aminokislot pri nizkih temperaturah. Razvedenie i kormlenie selskohozyaystvennyih zhivotnyih, 111–113 (in Russian).

Nauk, V. A. (1974). Izmeneniya pronitsaemosti membran i ustoychivosti belkovo-holosterinovyih kompleksov zhivchikov pod vliyaniem tehnologicheskiy obrabotki semeni byika. Selskohozyaystvennaya biologiya, 9(1), 102–108 (in Russian).

Nauk, V. A. (1974). O tselesoobraznosti otkaza ot “ekvilibratsii” pri zamorazhivanii semeni byikov. Zhivotnovodstvo, 10, 71–74 (in Russian).

Nauk, V. A. (1982). Kriokonservatsiya semeni zhivotnyih. Pushkino: Institut biologicheskoy fiziki (in Russian).

O’dell, W. T., & Almquist, J. O. (1957). Freezing bovine semen. I. Techniques for freezing bovine spermatozoa in milk diluents. Journal of Dairy Science, 40(12), 1534–1541. doi: 10.3168/jds.S0022-0302(57)94674-X.

O’dell, W. T., Almquist, J. O., & Marsh, L. A. (1958). Freezing bovine semen. III. Effect of freezing rate on bovine spermatozoa frozen and stored at − 79° C. Journal of Dairy Science, 41(1), 79–89. doi: 10.3168/jds.S0022-0302(58)90870-1.

Ostashko, F. I., & Buhrov, O. D. (1966). Pro hlyboke zamorozhuvannia spermy buhaiv-plidnykiv. Visnyk silskohospodarskoi nauky, 4, 86–91 (in Ukrainian).

Ostashko, F. I. (1959). Tryvale zberihannia simia buhaiv pry temperaturi – 183 ºC. Sotsialistychne tvarynnytstvo, 11, 36–39 (in Ukrainian).

Ostashko, F. I. (1964). Prichinyi i mehanizm holodovogo udara kletok. Mezhdunarodnyiy hozyaystvenney zhurnal, 3, 68–72 (in Ukrainian).

Ostashko, F. I., & Lopatko, M. I. (1965). Do pytannia pro zamorozhuvannia spermy plidnykiv. Zb. Rozvedennia i utrymannia silskohospodarskykh tvaryn, 5, 3–8 (in Ukrainian).

Ostashko, F. I., & Shinkarenko, V. A. (1972). Zamorazhivanie spermyi byikov-proizvoditeley v oblitsovannyih granulah. Sb. Tezisyi dokladov k nauchno-proizvodstvennoy konferentsii po teorii praktike vosproizvodstva selskohozyaystvennyih zhivotnyih. NII Zhivotnovodstva Lesostepi i Polesya USSR. Harkov, 54–55 (in Russian).

Ostashko, F. I. (1960). Novyiy metod zamorazhivaniya i dlitelnogo hraneniya semeni byikov pri temperature minus 1830S. Zhivotnovodstvo, 1, 79–84 (in Russian).

Ostashko, F. I. (1968). Glubokoe zamorazhivanie i dlitelnoe hranenie spermyi proizvoditeley. K.: Urozhay (in Russian).

Pakenas, P. I. (1963). Metodyi zamorazhivaniya semeni, primenyaemyie v Litovskim institute zhivotnovodstva i harakteristika potomstva, poluchennogo ot semeni perevezennogo iz Moskvyi. V kn. Novoe v plemennom dele i iskusstvennyim osemenenii selskohozyaystvennyih zhivotnyih, 201–207 (in Russian).

Petruk, E. M. (1966). Nekotoryie fiziologicheskie i morfologicheskie pokazateli spermiev pri glubokom zamorazhivanii spermyi byika (Doctoral dissertation, Lvovskiy zooveterinarnyiy institut) (in Russian).

Platov, E. M. (1960). Osmoticheskoe deystvie glitserina na zhivchikov byika. Vestnik selskohozyaystvennoy nauki, 11, 59–63 (in Russian).

Platov, E. M. (1965). Zamorazhivanie semeni barana v laktoz zheltochnoy-etilenglikolevoy srede. Ovtsevodstvo, 9, 11 (in Russian).

Platov, E. M. (1978). Nekotoryie teoreticheskie aspektyi zamorazhivaniya spermyi. Zhivotnovodstvo, 10, 64 (in Russian).

Platov E. M. (1982). Razrabotka i prakticheskoe primenenie tehnologii zamorazhivaniya spermyi baranov. Nauchno-praktich. konf. Po ovtsevodstvu i kozovodstvu: Tez. naukch. saobscheniy, 254–256 (in Russian).

Platov, E. M. (1963). Issledovaniya o deystvii glitserina PRI RAZBAVLENII I zamorazhivanii SEMENI selьskohozyaystvennyih zhivotnyih (Doctoral dissertation, Vsesoyuznaya ordena Lenina akademiya selskohozyaystvennyih nauk imeni VI Lenina; Vsesoyuznyiy ordena Trudovogo Krasnogo Znameni nauchno-issledovatelskiy institut zhivotnovodstva) (in Russian).

Platov, E. M. (1973). Teoreticheskie i prakticheskie osnovyi zamorazhivaniya semeni proizvoditeley selskohozyaystvennyih zhivotnyih. Diss.... d-ra biol. nauk. Dubrovitsyi (in Russian).

Platov, E. M., & Volkov, A. S. (1979). Effektivnost zamorazhivaniya spermyi baranov. Zhivotnovodstvo, 5, 53–56 (in Russian).

Pokhyl, V., Pokhyl, O., Honchar, A., & Lesnovska, O. (2011). Miasni porody ovetsu prydniprovi. Tvarynnytstvo Ukrainy, 9, 17–20 (in Ukrainian).

Polge, C., & Rawson, L. E. A. (1952). Longtery storage of bull semen froen at very low temperatures -79 °C. Proc 2-nd int. Cong. Anim. Reprod. Artif. Insem, 3, 90–98.

Polge, C. (1953). The preservation of spermatozoa at low temperatures. Mammalian germ cells, 6, 108.

Polge, C., & Rowson, L. E. A. (1952). Fertilizing capacity of bull spermatozoa after freezing at − 79° C. Nature, 169(4302), 626–627. doi: 10.1038/169626b0.

Polge, C., & Rowson, L. E. A. (1952). Results with bull semen stored at-79 C. Vet. Record, 64, 851.

Polge, C., Smith, A. U., & Parkes, A. S. (1949). Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, 164(4172), 666. doi: 10.1038/164666a0.

Pursel, V. G., Johnson, L. A., & Rampacek, G. B. (1972). Acrosome morphology of boar spermatozoa incubated before cold shock. Journal of Animal Science, 34(2), 278–283. doi: 10.2527/jas1972.342278x.

Rhodes, A. (1980). Semen collection and evaluation. Current therapy in theiogenology, 4, 944–947.

Roslanovskiy, K. (1963). Metodyi hraneniya semeni primenyaemyie v Polsche. V. kn.: Novoe v plemennom dele i iskusstvennom osemenenii selskohozyaystvennyih zhivotnyih. M. izd.s-h. literaturyi, zhurnalov i plakatov, 162–169 (in Russian).

Salamon, S. (1967). Observations on fertility of ram semen frozen by different methods. Australian Journal of Experimental Agriculture, 7(29), 559–561. doi: 10.1071/EA9670559.

Salamon, S. (1968). Deep freezing of ram semen: recovery of spermatozoa after pelleting and comparison with other methods of freezing. Australian journal of biological sciences, 21(2), 351–360. doi: 10.1071/BI9680351.

Salamon, S., & Visser, D. (1972). Effect of composition of Tris-based diluent and of thawing solution on survival of ram spermatozoa frozen by the pellet method. Australian journal of biological sciences, 25(3), 605–618. doi: 10.1071/BI9720605.

Schmidt, D., Peter, W., Mudra, K., & Kunze, C. (1974). Untersuchungen zur Gefrierkonservierung von Ebersperma. 3. Prufung der Befruchtungsfahigkeit. Archiv fur Tierzucht.

Semekov, V. G. (1984). Soderzhanie tsiklicheskih nukleotidov (TsAMF i IDGMF) v sperme byikov i hryakov pri kriokonservatsii. Selskohozyaystvennaya biologiya, (2),81–83 (in Russian).

Shaydullin, I. N. (1980). Usovershenstvovanie metoda zamorazhivaniya semeni barana putem primeneniya antioksidantov. Avtoreferat diss. kand. biol. nauk (in Russian).

Shergin, N. P. (1967). Biohimiya spermyi selsko-hozyaystvennyih zhivotnyih. M.: Kolos (in Russian).

Skornyakov, B. A. (1973). Izuchenie povrezhdenii spermiev byika pri ih nizkotemperaturnoy konservatsii. Avtoreferat kand. diss (in Russian).

Smirnov, I. V. (1951). Sohranenie semeni spermyi selskohozyaystvenyih zhyivotnyih pri temperature -78 °С. Sovetskoe zhyivotnovodstvo, 1, 94–95 (in Russian).

Smirnov, I. V. (1963). Vliyanie glitserina i gipertonicheskih rastvorov na prizhivaemost spermiev byikov-proizvoditeley. Nauchnyie trudyi Kievskoy opyitnoy stantsii zhivotnovodstva, 9, 43–52 (in Russian).

Smirnov, I. V. (1964). Deystvie osmoticheskih faktorov na spermii byikov i hryakov, 85–93 (in Russian).

Smirnov, I. V. (1949). Sohranenie semeni selsko-hozyaystvennyih zhivotnyih posredstvom glubokogo ohlazhdeniya. Sovetskaya zootehniya, 4, 93–96 (in Russian).

Smirnov, I. V. (1974). K teorii glubokogo ohlazhdeniya spermyi. Zhivotnovodstvo, 11, 65–70 (in Russian).

Sokolovskaya, I. I., Oyvadis, R. N., Abilov, A. I. i dr. (1981). O znachenii okrasom v otsenke semeni samtsov. Zhivotnovodstvo, 9, 46–47 (in Russian).

Varnavskiy A. N. (1969). Ultrastruktura zhivchikov byika i barana posle glubokogo zamorazhivaniya v zhidkom azote. Selskohozyaystvennaya biologiya, 3, 469–471 (in Russian).

Varnavskiy, A. N. (1970). Vozmozhnaya prichina snizheniya oplodotvoryayuschey sposobnosti zamorozhennogo semeni. Ovtsevodstvo, 2, 28–29 (in Russian).

Varnavskiy, A. N. Turbin, V. F., & Osadchuk, V. S. (1972). Podvizhnost i ultrastruktura zhivchikov byika v protsesse tehnologicheskoy obrabotke. Tehnologiya iskusstvennogo osemeneniya i biologiya vosproizvedeniya selskohozyaystvennyih zhivotnyih, 169–175 (in Russian).

Varnivskiy, A. N. (1978). Izuchenie akrasom spermatozoidov baranov s pomoschyu fazokontrastnoy mikroskopii. Dokladyi VASHNIL, 8, 33–34 (in Russian).

Visser, D., & Salamon, S. (1974). Fertility following inseminations with frozen-thawed reconcentrated and unconcentrated ram semen. Australian journal of biological sciences, 27(4), 423–426. doi: 10.1071/BI9740423.

Volkov, A. S. (1971). Izuchenie oplodotvoryaemoy ovets pri osemenenii zamorozhennyim semenem barana. Avtoreferat kand. diss. Dubrovitsyi (in Russian).

Volkov, A. S. (1974). Vliyanie chisla aktivnyih zhivchikov v ottayannom semeni na oplodotvoryaemost ovets. Zhivotnovodstvo, 10, 69–71 (in Russian).

Voronenko, V., Iovenko, V., & Zharuk, P. (2009). Henetychni resursy ovets Ukrainy ta umovy yikh podalshoho rozvytku. Naukovyi visnyk Natsionalnoho universytetu bioresursiv i pryrodokorystuvannia Ukrainy, 287–296 (in Ukrainian).

Watson, P. (1975). Use of a Giemsa stain to defect changes in aerosomes of frozen ram spermatozoa. Veter. Res., 97(1), 12–15. doi: 10.1136/vr.97.1.12.

Watson, P. F. (1981). The effect cold shock on sperm cell membrane. Effect of Low temperatures on the biological membranes, 198–218.

Watson, P. F. (1975). Use of a Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa. The Veterinary Record, 97(1), 12. doi: 10.1136/vr.97.1.12.

White, I. D., Blackshaw, A. W., & Emmens, C. W. (1954). Metabolic and motility studies relating to the low temperature storage of ram and bull spermatozoa. Australian Veterinary Journal, 30(4), 85–94. doi: 10.1111/j.1751-0813.1954.tb08161.x.

Wiggin, H. B., & Almquist, J. O. (1975). Effect of glycerol equilibration time and thawing rate upon acrosomal maintenance and motility of bull spermatozoa frozen in plastic straws. Journal of animal science, 40(2), 302–305. doi: 10.2527/jas1975.402302x.

Wilmut, I., & Polge, C. (1977). The low temperature preservation of boar spermatozoa. 1. The motility and morphology of boar spermatozoa frozen and thawed in the presence of permeating protective agents. Cryobiology, 14(4), 471–478. doi: 10.1016/0011-2240(77)90009-8.

Yatsun, A. S., & Ilinskaya, T. P. (1969). O pronitsaemosti glitserina cherez obolochku spermiev. V sb. Aktualnyie voprosyi povyisheniya produktivnosti selskohozyaystvennyih zhivotnyih. Minsk, Urozhay, 75–78 (in Russian).

Yuschenko, N. P., Semakov, V. G., & Levin, K. A. (1963). Zamorazhivanie spermyi byikov v granulah bez primeneniya suhogo lda. Molochnoe i myasnoe skotovodstvo, 5, 36–37 (in Russian).

Zheltobryuh, N. A., & Ivahnenko, V. K. (1977). Primeneniya saharno-zheltochnyih sred dlya zamorazhivaniya spermyi barana. Tr. VNIIOK, 39(1),111–118 (in Russian).

Zheltobryuh, N. A., & Ivahnenko, V. K. (1977). Sovershenstvovanie tehnologii zamorazhivaniya i ottaivaniya spermyi barana. Mat. nauchno-proizv, 27–28 (in Russian).

Zhiltsov, N. Z. (1970). Kontsentratsiya kisloroda v kanale epididimisa krolika. Dokladyi VASHNIL, 5, 34–35 (in Russian).

Zhiltsov, N. Z., Golub, V. S., Varnavskaya, V. A. i dr. (1974). O vozmozhnosti primeneniya prostaglandinov dlya povyisheniya effektivnosti osemeneniya ovets. Zhivotnovodstvo, 8, 67–70 (in Russian).

Zhiltsov, N. Z. (1999). Novoe v biologii vosproizvedeniya selskohozyaystvennyih zhivotnyih. Zootehniya, 11, 31 (in Russian).

Zvereva, A. P. (1976). Sovershenstvovanie tehnologii zamorazhivaniya spermyi byikov-proizvoditeley i ispyitanie novyih krioprotektorov. Avtoreferat kand. dis (in Russian).

Zvereva, A. P. (1977). Vliyanie protsessov adaptatsii i ekvilibratsii na biologicheskie svoystva spermiev byikov pri zamorazhivanii. Fiziologiya vosproizvodstva selskohozyaystvennyih zhivotnyih, 229–232 (in Russian).

Zvereva, G. V., & Chernomaz, L. A. (1977). Nekotoryie voprosyi morfologii spermiev byika. V kn. Novoe v plemennom dele i iskusstvennom osemenenii selskohozyaystvennyih zhivotnyih, 378–388 (in Russian).

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Published
2020-10-06
How to Cite
Hrymak, K. (2020). Cryopreservation of ram-sires semen and its qualitative changes under the influence of low temperatures. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Agricultural Sciences, 22(93), 102-112. https://doi.org/10.32718/nvlvet-a9318