تأثیر مورفومتری، ویسکوزیته و آبکی‌شدن منی بر کیفیت اسپرم منجمد قوچ قزل

نوع مقاله : مقاله علمی پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد فیزیولوژی دام، گروه علوم دامی دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.

2 استاد گروه علوم دامی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.

چکیده

توانایی اسپرم در حرکت و رسیدن به اووسیت جهت انجام لقاح و باروری به عوامل مختلفی از جمله مورفومتری، آبکی­شدن و ویسکوزیته‌ منی بستگی دارد. مطالعهء حاضر به منظور ارزیابی تأثیر مورفومتری، ویسکوزیته و آبکی‌شدن مایع منی بر کیفیت اسپرم منجمد قوچ قزل انجام گرفت. جمع­آوری منی یکبار در هفته به مدت 3 هفته از 5 رأس قوچ قزل صورت گرفت. ارزیابی­های اولیه شامل تحرک کل و پیشرونده، درصد زنده‌مانی، درصد اسپرم‌های غیرطبیعی، غلظت، ویسکوزیته، مورفومتری و آبکی‌شدن بود. نمونه‌های با غلظت بالای 5/2 میلیارد اسپرم و حرکت پیشروندهء بالای 70 درصد برای رقیق‌سازی مورد استفاده قرار گرفتند. بعد از رقیق­سازی، نمونه‌ها در داخل پایت‌های 25/0 میلی­لیتری کشیده شده و در یخچال به مدت 2-5/1 ساعت جهت سردسازی تا دمای 5 درجهء سلسیوس قرار گرفتند. سپس به مدت 10-8 دقیقه در 5-4 سانتی­متری بالای سطح نیتروژن مایع در بخار ازت قرار گرفتند و در نهایت داخل نیتروژن مایع غوطه­ور شدند. یخ‌گشایی نمونه‌ها در روزهای صفر، 20، 40 و 60 دوره نگه­داری، انجام شد. نتایج نشان داد که صفات مربوط به تحرک و زنده‌مانی اسپرم­ها با گذشت زمان کاهش معنی‌داری داشتند (05/0>p). بین طول قطره و یکپارچگی غشای پلاسمایی منی هم رابطهء منفی و معنی‌دار وجود داشت (05/0>p). همچنین بین طول رشتهء منی و تحرک کل، همبستگی مثبت و معنی‌دار، ولی بین آبکی‌شدن با تحرک کل، حرکت پیشرونده، زنده‌مانی و یکپارچگی آکروزوم، همبستگی منفی و معنی‌دار مشاهده گردید (01/0>p). بین طول دم و طول کل اسپرم با یکپارچگی غشای پلاسمایی نیز رابطهء منفی و معنی‌دار مشاهده شد (01/0>p). یافته­های تحقیق حاضر مشخص کرد که هرچه زمان آبکی‌شدن مایع منی، طول دم و طول کل اسپرماتوزئید کوتاه‌تر باشد، صفات کیفی آن در فرایند انجماد بهتر حفظ خواهد شد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

The effect of morphometry, viscosity and liquefaction on frozen sperm quality of Ghezel ram

نویسندگان [English]

  • parisa shafaati 1
  • gh moghaddam 2
1 MSc Student, Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran.
2 Professor, Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran.
چکیده [English]

The ability of sperm to move and reach the oocyte for fertilization and fertility depends on different factors such as morphometry, liquefaction and semen viscosity. The present study was performed to evaluate the effect of morphometry, viscosity and liquefaction of semen on Ghezel ram frozen sperm quality. Semen was collected once a week for 3 weeks from 5 Ghezel rams. Initial assessments included total and progressive motility, viability, abnormal sperm percentage, concentration, viscosity, morphometry, and liquefaction. Samples with a concentration of 2.5 billion sperm and a progressive motility of over 70% were used for dilution. After dilution, straws (0.25 ml) were filled and after cooling and reaching the temperature of 5°C in the refrigerator, were placed 4-5 cm above liquid nitrogen for 8-10 min and then, ultimately were immersed in liquid nitrogen. The traits of motility and sperm health were assessed on days 0, 20, 40 and 60 of cryopreservation. The results showed that motility traits and viability were significantly reduced over time of cryopreservation (p<0.05). Furthermore, the results of this study showed a negative and significant correlation between drop length and plasma membrane health (p<0.05). Also, it was shown that there was a positive and significant correlation between semen string length and total motility (p<0.01). There was a significant negative correlation between liquefaction and total motility, progressive motility, viability and acrosome integrity (p<0.01). There was a significant negative correlation between tail length and total sperm length with plasma membrane health (p<0.01). The findings of the present study indicated that the shorter the liquefaction time, sperm tail length and total length of sperm, the qualitative traits of sperm will be better preserved in the freezing process.

کلیدواژه‌ها [English]

  • cryopreservation
  • Liquefaction
  • Morphometry
  • sperm
  • Viscosity
  • Amelar, R.D. (1962). Coagulation, liquefaction and viscosity of human semen. Journal of Urology, 87(2): 187-190.
  • Anamthathmakula, P. and Winuthayanon, W. (2020). Mechanism of semen liquefaction and itspotential for a novel non-hormonalcontraception. Biology of Reproduction, 103(2): 411-426.
  • Ansari, M., Tohidi, A. and Mohammad Moradishahr, B. (2011). Evaluation the effect of adding different levels of n-3 fatty acids to bioxel diluent on the freezing capacity of goat sperm. The Journal of Veterinary Clinical Pathology, 4(4): 1013-1017. [In Persian]
  • Berns, M.W., Chandsawangbhuwana, C., Hyun, N., Shi, L.Z., Yang-Wong, C. and Zhu, O. (2012). Effects of viscosity on sperm motility studied with optical tweezers. Journal of Biomedical Optics, 17(2): 1-6.
  • Breed, W.G. (1983). Variation in sperm morphology in the Australian rodent genus, Pseudomys (Muridae). Cell and Tissue Research, 229(3): 611-625.
  • Chaudhuri, N., Nag, A. and Nag, P.K. (1979). Relative Viscosity of Human Seminal Fluid: Influence of Sperm Concentration, Motility and Biochemical Ingredients. Andrologia, 11(6): 478-482.
  • Cunha, D., Brito, B., Evangelista, J., Júnior, F., Pereira, L. Ribeiro, L., et al. (2020). Characterization of seminal parameters, sperm morphometry, micromorphology, and ultrastructure in gray brocket deer (Mazama gouazoubira, Fischer, 1814). Microscopy Research Technology, 84(2): 313-325.
  • Dresdner, R.D. and Katz, D.F. (1981). Relationships of mammalian sperm motility and morphology to hydrodynamic aspects of cell function. Biology of Reproduction, 25(5): 920-930.
  • Elzanaty, S., Malm, J. and Giwercman, A. (2004). Visco-elasticity of seminal fluid in relation to the epididymal and accessory sex gland function and its impact on sperm motility. International Journal of Andrology, 27(2): 94-100.
  • Evans, J.P., Fitzpatrick, J.L., Humphries, S., Simmons L.W. and Simpson, J.L. (2012). Relationship between sperm length and sperm differ among three internally and externally fertilizing species. Journal Storage, 68(1): 92-104.
  • Gadea, J., Molla, M., Selles, E., Marco, M.A., Garcia-Vazquez, F.A. and Gardon, J.C. (2011). Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology, 62(1): 40-46.
  • Garcia‑Vazquez, F.A., Hernandez‑Caravaca, I., Matas, C., Soriano‑Ubeda, C. and Abril‑Sanchez, S. (2015). Morphological study of boar sperm during their passage through the female genital tract. Journal of Reproduction and Development, 61(5): 407-413.
  • Hernandez‑Caravaca, I., Soriano‑Ubeda, C., Matas, C., Izquierdo‑Rico, M.J. and Garcia‑Vazquez, .F.A. (2015). Boar sperm with defective motility are discriminated in the backflow moments after insemination. Theriogenology, 83(4): 655-661.
  • Hosseini, S., Mohammad khani, A.G., Moghbelinejad, S. and Shabani, Kh. (2017). The effects of Semen Parameters and age on Sperm Motility of Iranian men. Global Journal of Fertility and Research, 2(1): 024-029.
  • Karagiannidis, A., Alexopoulos, C., Amarantidis, I. and Varsakeli, S. (2000). Seasonal variation in semen characteristics of Chois and Feresian rams in Greece. Small Ruminant Research, 37(1-2): 125-130.
  • Keel, B.A. (1990). The semen analysis. In: Keel, B., Webster, B., editors. CRC Handbook of the laboratory diagnosis and treatment of infertility. Boca Raton: CRC Press, pp: 27-69.
  • Lupold, S. and Fitzpatrick, J.L. (2015). Sperm number trumps sperm size in mammalian ejaculate evolution. Proceedings of the Royal Society, 282(1819): 1-7.
  • Moghaddam, G.H., Pourseif, M.M. and Rafat, S.A. (2012). Seasonal variation in semen quantity and quality traits of Iranian crossbred rams. Slovak Journal of Animal Science, 45(3): 67-75.
  • Moon, K.H. and Bunge, R.G. (1968). Observations on the biochemistry of human semen. III. Amylase. Fertility and Sterility, 19(6): 977-981.
  • Ozkavukcu, S., Erdemli, E., Isik, A., Oztuna, D. and Karahuseyinoglu, S. (2008). Effects of cryopreservation on sperm parameters and ultrastructural morphology of human spermatozoa. Journal of Assisted Reproduction and Genetics, 25(8): 403-411.
  • Palacin, I., Alquezar-Baeta, C., Santolaria, P., Silvestre, M.A., Soler, C. and Yaniz, J. (2020). Relationship of sperm plasma membrane and acrosomal integrities with sperm morphometry in Bos­taurus. Asian Journal of Andrology, 22(6): 578-582.
  • Rastegharnia, A. and Shafipour, V. (2008). The effect of lactose, milkweed and tris diluents on buffalo frozen sperm. The Journal of Veterinary Clinical Pathology, 1(2): 79-88. [In Persian]
  • Santolaria, P., Vicente‑Fiel, S., Palacin, I., Fantova, E. and Blasco, M.E. (2015). Predictive capacity of sperm quality parameters and sperm subpopulations on field fertility after artificial insemination in sheep. Animal Reproduction Science, 163(2015): 82-88.
  • Shafaati Alishah, P., Moghaddam, G.h., Daghighkia, H. and Alijani, S. (2020). Investigating the effect of diluents containing EDTA and Propylene glycol on survival of frozen semen of Ghezel ram.  The Journal of Veterinary Clinical Pathology, 52(13): 353-369. [In Persian]
  • Silva, S., Soares, A., Batista, A., Almeida, F., Nunes, J., Peixoto, C. and Guerra, M. (2013). Vitamin E (Trolox) addition to Tris-egg yolk extender preserves ram spermatozoa structure and kinematics after cryopreservation. Animal Reproduction Science, 137(1-2): 37-44.
  • Vasan, S.S. (2011). Semen Analysis and sperm function tests: How much to test. Indian Journal of Urology, 27(1): 41-48.
  • World Health Organization. (2010). WHO laboratory manual for the examination and processing of human semen. 5th ed., World Health Organization, pp: 140-143.
  • Zanganeh, Z., Mohammadi-Sangcheshmeh, A., Nabi, M.M., Najafi, A., Zare-Shahneh, A. and Zhandi, M. (2013). Does rosemary aqueous extract improve buck semen cryopreservation? Small Ruminant Research, 114(1): 120-125.