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УДК: 639.212:591.391 DOI:10.33920/sel-09-2105-05

Hypothermic storage in salt-free preservative solution alter motility duration in sterlet sperm

Dmitry Alexandrovich Isaev PhD, senior research scientist, All-Russian research Institute of integrated fish farming, a branch of the Federal state budgetary scientific institution “Federal research center for animal husbandry — VIZ named after academician L.K. Ernst”, Russia, 142460, Moscow Region, Noginsk District, Vorovsky village, Sergeev Str, 24, E-mail: dmais@hotmail.ru, ORCID: 0000-0001-6706-8606
Alexander Pavlovich Glebov chief fish production officer, Central Branch of the Federal State-financed Institution “Head Basin Administration for Fisheries and Conservation of Aquatic Biological Resources”, Russia, 17105, Moscow, Varshavskoe shosse, 39A, E-mail: glebov74@rambler.ru, ORCID: 0000-0002-1570-3839
Marina Yurievna Martynova PhD, associate professor, Moscow Medical University “Reaviz”, Russia, 107564, Moscow, Krasnobogatyrskaya Str, 2, build. 2, entr. 22, E-mail: mm122@yandex.ru, ORCID: 0000-0001-6911-619X
Elena Ivanovna Shishanova PhD, director, All -Russian research Institute of integrated fish farming, a branch of the Federal state budgetary scientific institution “Federal research center for animal husbandry — VIZ named after academician L.K. Ernst”, Russia, 142460, Moscow Region, Noginsk District, Vorovsky village, Sergeev Str, 24, E-mail: lenavniir@mail.ru, ORCID: 0000-0001-6158-1871

Together with the concentration, the motility of sturgeon sperm is one of the most important characteristics determining its quality and suitability for fertilization. The duration of forward movement of spermatozoa after activation in water is also important, and this time should not be less than necessary for fertilization. The motility of spermatozoa depends on their physiological state, degree of maturity, age, and intracellular reserves of high-energy compounds. The duration of sperm motility is influenced by factors such as osmolality, the ionic composition of the medium, pH, and temperature. During hypothermic storage of sperm, the number of spermatozoa capable of activation progressively decreases due to depletion of ATP reserves or death. To improve the hypothermic storage of sterlet sperm, we developed a salt-free based on glucose and trehalose preserving solution ISGT-80. When sterlet sperm samples from 20 males were stored in ISGT80 for 18 days, we observed, along with a progressive decrease in the number of motile spermatozoa, a change in the duration of their motility. On average, the motility half-loss time (τ50) on the 3rd-6th days of storage significantly increased by about 1 minute compared with fresh samples, then gradually decreased, however, without falling to the initial value. The reasons for this prolongation of motility are not clear, but we do not exclude the preferential death in the first days of storage of spermatozoa with a short period of motility and the selection in favor of "long-living" spermatozoa. Such selection at the gamete level can change in allele frequencies of heterozygous loci in the offspring. Thus, hypothermic sperm storage can be an interesting subject of genetic research to develop new selection tools in sturgeon breeding.

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Mature spermatozoa of sturgeon fish in the testes and seminal ducts are immotile as long as they contact tissue fluids or seminal plasma [14; 15]. During spawning, spermatozoa in contact with freshwater are activated and become motile. For most sturgeon fish, the active motility time does not exceed 2-5 minutes, and the total motility time does not exceed 10-15 minutes [15]. Together with concentration, motility is one of the main characteristics of sperm quality and its suitability for artificial insemination [11]. Despite this, the relationship between motility duration and sperm fertility is not evident because all whole eggs capable of fertilization are inseminated within the first 30-60 seconds [11]. N.V. Barulin et al. [1], using CASA (computer assisted sperm analyses) to study motility and other characteristics of Lena sturgeon sperm, proved the inexpediency of artificial insemination for more than 90 seconds.

The motility of sturgeon sperm depends on many factors [16]. But in the case of short-term or long-term sperm storage technologies, such as hypothermic storage or cryopreservation, sperm is additionally exposed to non-specific factors (cryoprotectants, antibiotics, etc.) or environmental factors beyond the physiological ranges (low temperatures, pH, differences in osmotic pressure, etc.) that affect not only sperm viability, but also motility patterns [26-28].

Despite a large number of developments, cryopreservation of sturgeon sperm is not used as a standard technology in sturgeon breeding [7]. Much more often, short-term hypothermic storage without freezing, at near-zero temperatures, is used for transport or delayed insemination of eggs. The method of hypothermic storage of sturgeon sperm recommended by FAO [11] is the Bijar-DiLauro method: in plastic (polyethylene) bags or containers filled with atmospheric air or pure oxygen [20; 23; 24].

Hypothermic storage decreases sperm motility as a result of progressive death of spermatozoa or loss of their ability to activate. The main causes are depletion of ATP reserves, lipid peroxidation, and, consequently, destruction of cell membranes [19], spontaneous premature activation and subsequent death of spermatozoa; microbial overgrowth due to inevitable contamination is also possible [5]. To improve the preservation of sterlet sperm during hypothermic storage in 2012-2016, we developed a salt-free medium ISGT-80 based on glucose, trehalose, and bovine serum albumin with osmolality similar to that of sterlet seminal plasma. After storage in ISGT-80 at +2-4 °C, at least 50% of sterlet sperm became motile after activation for 6 days. The fraction of spermatozoa with damaged DNA after 6 days of storage did not exceed 1.5%, and spermatozoa with damaged membranes did not exceed 10% [8]. When sterlet eggs were inseminated with sperm stored for 10 days in ISGT-80, the fertilization rate was at least 90% [10].

Для Цитирования:
Dmitry Alexandrovich Isaev, Alexander Pavlovich Glebov, Marina Yurievna Martynova, Elena Ivanovna Shishanova, Hypothermic storage in salt-free preservative solution alter motility duration in sterlet sperm. Рыбоводство и рыбное хозяйство. 2021;5.
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