بررسی صفات اسپرم رقیقشده قوچ با اسانس آویشن (Thymus vulgaris)، با استفاده از میکروسکوپ فازکنتراست و نرمافزار CASA
محورهای موضوعی :
آسیب شناسی درمانگاهی دامپزشکی
حدیثه قمری منور
1
(دانشجوی دکترای گروه علوم دامی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.)
غلامعلی مقدم
2
(استاد گروه علوم دامی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.)
حسین دقیق کیا
3
(استاد گروه علوم دامی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.)
بابک قاسمی پناهی
4
(استادیار گروه علوم دامی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران.)
تاریخ دریافت : 1401/12/01
تاریخ پذیرش : 1402/05/14
تاریخ انتشار : 1402/02/01
کلید واژه:
آویشن,
اسپرم,
قوچ,
صفات حرکتی,
نرمافزار CASA,
چکیده مقاله :
توانایی اسپرم در انجام لقاح به صفات مختلفی ازجمله حرکت، زنده مانی و حجم کافی بستگی دارد. پژوهش حاضر بهمنظور بررسی تاثیر اسانس آویشن بر صفات حرکتی اسپرم قوچ با استفاده از میکروسکوپ فاز کنتراست و نرمافزار CASA (Computer Assisted Sperm Analyzer) انجام گرفت. بدین منظور اسپرم گیری از 4 رأس قوچ و هفته ای دوبار انجام شد. ارزیابی اولیه شامل حجم، حرکت موجی و پیش رونده، حرکت درجا و درصد زنده مانی اسپرم ها بود. سپس نمونه های اسپرم با رقیق کننده شاخص استاندارد بر پایه تریس، 20 درصد زرده تخم مرغ، 7 درصد گلیسرول، به علاوه مقادیر صفر، 100، 200 و 400 میکروگرم اسانس آویشن، مخلوط شدند. بعد از رساندن دمای نمونه های اسپرم به 5 درجه سلسیوس در یخچال، به مدت 10 دقیقه در فاصله 4 سانتی متری بالای نیتروژن مایع قرار گرفتند و در نهایت داخل آن غوطه ور شدند. نمونه ها در روزهای صفر و 30 آزمایش، یخ گشایی شدند. در روزهای انجماد و ذوب، تحرک کل و پیش رونده، حرکت درجا، درصد زنده مانی و صفات CASA و مقدار مالون دی آلدئید (malondialdehyde) مورد آزمایش قرار گرفتند. نتایج نشان داد که درصد زنده مانی و تحرک و مقدار MDA در طول دوره آزمایش کاهش معنی داری داشتند (001/0>p) و تیمارهای 100، 200 و 400 میکروگرم آویشن، بیشترین اختلاف معنی دار را نسبت به گروه شاهد در صفات حرکت پیش رونده و کل، زندهمانی و CASA به خود اختصاص دادند (001/0>p). در صفت MDA تیمارهای 100 و 200 اختلاف معنیداری نسبت به تیمار 400 و شاهد داشتند (001/0>p). نتایج پژوهش حاضر نشان داد که استفاده از اسانس آویشن به عنوان رقیقکننده باعث بهبود صفات حرکتی اسپرم و زنده مانی در طول انجماد می شود.
چکیده انگلیسی:
The ability of sperm to perform fertilization depends on various characteristics such as movement, viability, and sufficient volume. The present study was conducted to investigate the effect of Thymus vulgaris essential oil on the motility characteristics of ram sperm using phase-contrast microscope and CASA (Computer Assisted Sperm Analyzer) software. For this purpose, semen was collected twice a week from four rams. Initial evaluations included volume, wave motility, progressive motility, non-progressive motility and survival percentage of sperms. Then sperm samples were mixed with standard index Tris-based diluent, 20% egg yolk, 7% glycerol, plus 0, 100, 200 and 400 µg of Thymus vulgaris essential oil per ml. After bringing the temperature of the sperm samples to 5 degrees Celsius in the refrigerator, they were placed at 4 cm above liquid nitrogen for 10 minutes and finally immersed in it. The samples were thawed on days 0 and 30 of the experiment. Traits which were investigated during freezing and thawing days included total and progressive motility, non-progressive motility, viability percentage, MDA (Malondialdehyde) and CASA characteristics. The results showed that the percentage of viability and mobility decreased significantly during the experiment (p<0.001). The treatments of 100, 200 and 400 µg Thymus vulgaris essential oil had the most significant percentages compared to the control group in the characteristics of progressive and total motility, viability and CASA (p<0.001). Malondialdehyde had significantly decreased in the 100 and 200 treatments compared to the 400 and control treatments (p<0.001). The results of the present study showed that diluents containing Thymus vulgaris essential oil improve sperm motility and viability during freezing.
منابع و مأخذ:
Ahmed, N.A.H., Nahed, M. and El-Aal, R.A. (2015). Assessing the effect of thyme and rosmary as antiaflatoxicosis on fertility in male rats. The Journal of American Science, 11(12): 294-302.
Alasadiy, Y.D.K. (2014). Study of the biological effect of Thymus vulgaris extracts on spermatogenesis in experimentally infected white mice Balb/c by Toxoplasma gondii. International Journal of Scientific and Engineering Research, 5(10): 547-553.
Ariza-Nieto, C., Bandrick, M., Baidoo, S.K., Anil, L., Molitor, T.W. and Hathaway, M.R. (2011). Effect of dietary supplementation of oregano essential oils to sows on colostrum and milk composition, growth pattern and immune status of suckling pigs. Journal of Animal Science, 89(4): 1079-1089.
Bailey, J.L., Blodeau, J.F. and Cormier, N.Y. (2000). Semen cryopreservation in domestic animals: A damaging and capacitating phenomenon mini Journal of Andrology, 21(1): 1-7.
Bittencourt, R.F., Bicudo, S.D., Biscarde, E.A., Chalhoub, M., Filho, A.D.L. and Oba, E. (2014). Trehalose and Calcium chelator for ram semen cryopreservation. Archives of Verterinary Science, 19(2): 66-77.
Chikhoune, A., Stouvenel, L., Iguer-Ouada, M., Hazzit, M., Schmitt, A., Lores, P., et al. (2015). In-vitro effects of Thymus munbyanus essential oil and Thymol on human sperm motility and function. Reproductive Biomedicine Online, 31(9): 411-420.
Cunha, D., Brito, B., Evangelista, J., Junior, F., Pereira, L., Rebeiro, 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.
Daghigh kia, H., Sadeghi, F., Ebrahimi, M. and Samadian, F. (2017). Comparative effect of different concentrations of hydro-ethanolic extract of chamomile on freeze-thawn semen quality of rams. Journal of Veterinary Clinical Pathology, 41(11): 13-24. [In Persian]
Daghigh Kia, H., Farhadi, R., Ashrafi, I. and Mehdipour, M. (2016). Anti-oxidative effects of ethanol extract of Origanum vulgare on kinetics, microscopic and oxidative parameters of cryopreserved Holstein bull spermatozoa. Iranian Journal of Applied Animal Science, 6(4): 901-907.
Eriksson, B.M. and Rudriguez-Martinez, H. (2000). Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa frozen in Flat Packs and Maxi-Straws. Animal Reproduction Science, 63(3-4): 205-220.
Esterabauer, H. and Cheeseman K.H. (1990). Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods in Enzymology, 186(3): 407-421.
Gedicoglu, A., Sokmen, M. and Civit, A. (2019). Evaluation of Thymus vulgaris and Thymbra spicata essential oils and plant extracts for chemical composition, antioxidant and antimicrobial properties. Food Science and Nutrition, 7(5): 1704-1714.
Hamzawy, M.A., El-Denshary, E.S.M., Hassan, N.S., Manaa, F. and Abdel Wahhab, M.A. (2012). Antioxidant and renoprotective effects of Thymus vulgaris extracts in rats during aflatoxicosis. Global Journal of Pharmacy and Pharmacology, 6(2): 106-117.
Hazzit, M., Baaliouamer, A., Faleiro, M.L. and Miguel, M.G. (2006). Composition of the essential oils of Thymus and Origanum species from Algeria and their antioxidant and antimicrobial activities. Journal of Agricultural and Food Chemistry, 54(17): 6314-6321.
Hu, J.H., Chen, X.Y., Li, G., Li, Q.W., Yang, H., Wang, L.I., et al. (2006). The cryopreservation effect on frozen-thawed semen of egg yolk low density lipoproteins. Journal of Animal Science, 19(4): 486- 494.
Ieropoli, S., Do Espirito Santo, M., Masullo, P. and Sansone, G. (2004). Effect of extender composition, cooling rate and freezing on the fertilization viability of spermatozoa of the Pacific oyster (Crassostrea gigas). Cryobiology, 49(3): 250-257.
Jalili, C., Kamani, M., Roshankhah, S., Sadeghi, H. and Salahshoor, M.R. (2018). Effect of Falcaria vulgaris extracts on sperm parameters in diabetic rats. Andrologia, 50(10): 1-9.
Lupold, S. and Fitzpatrick, J.L. (2015). Sperm number trumps sperm size in mammalian ejaculate evolution. Proceedings of the Royal Society, 282(1): 1-7.
Maia, J.D., La Corte, R., Martinez, J., Ubbink, J. and Prata, A.S. (2019). Improved activity of Thyme essential oil (Thymus vulgaris) against Aedes aegypti larvae using a biodegradable controlled release system. Industrial Crops and Products, 136(2): 110-120.
Malo, C., Gil, L., Cano, R., Martinez, F. and Gale, I. (2011). Antioxidant effect of rosmary (Rosmarinus officinalis) on boar epididymal spermatozoa during cryopreservation. Theriogenology, 75(9): 1735-1741.
Masoudi, R., Shahneh, A.Z. and Asadzadeh, N. (2019). Improvement of ram’s frozen sperm quality using Glutathione antioxidant. Veterinary Research and Biological Products, 33(128): 69-75. [In Persian]
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.
Nateq, S., Moghaddam, G.H., Alijani, S. and Behnam, M. (2020). The effects of different levels of Nano selenium on the quality of frozen-thawed sperm in ram. Journal of Applied Animal Reaserch, 48(1): 434-439.
Novak, J., Bitsch, C., Langbehn, J., Pank, F., Skoula, M., Gotsiou, Y., et al. (2000). Ratios of cis- and trans-sabinene hydrate in Oryganum majorana L. and Origanum microphyllum (Bentham) Vogel. Biochemical Systematics and Ecology, 28(7): 697-704.
Ozguven, M. and Tansi, S. (1998). Drug yield and essential oil of Thymus vulgaris L. as in influenced by ecological and ontogenetical variation. Turkish Journal of Agriculture and Forestry, 22(6): 537- 542.
Salahshoor, M.R., Abdolmaleki, A., Faramarzi, A., Ziapour, A. and Roshankhah, S. (2020). Thymus vulgaris attenuates Myleran-induced reproductive damage by decreasing oxidative stress and Lipid peroxidation in male rats. Journal of Human Reproductive Sciences, 13(1): 38-45.
Shanoon, A.K. and Mahdi, S.J. (2012). Effects of Thymus vulgaris and Zingiber officinale aqueous on semen parameters, testes weight and histology measurement of broiler breeder male. International Journal of Poultry Science, 11(9): 594-598.
Shafaati, P. and Moghaddam, G.H. (2021). The effect of morphometry, viscosity and liquefaction on frozen sperm quality of Ghezel ram. Journal of Veterinary Clinical Pathology, 58(15): 155-167. [In Persian]
Shafaati, P., Moghaddam, G.H., Daghigh kia, H. and Alijani, S. (2020). Investigating the effect of diluents containing EDTA and Propylene glycol on survival of frozen semen of Ghezel ram. Journal of Veterinary Clinical Pathology, 52(13): 353-369. [In Persian]
Sharafi, M.M., Forouzanfar, S.M., Hosseini, M., Hajian, M., Ostadhosseini, S., Hosseini, L., et al. (2009). In vitro comparison of soybean lecithin based- extender with commercially available extender for ram semen cryopreservation. International Journal of Fertility and Sterility, 3(3): 149-152.
Sicherle, C.C., Maia, M.S., Bicudo, S.D., Rodello, L. and Azqvedo, H.C. (2011). Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen supplemented with catalase or Torolox. Small Ruminant Research, 95(2-3): 144-149.
Silva, S.V., Soares, A.T. Batista, A.M., Almeida, F.C., Nunes, J.F. and Peixoto, C.A. (2013). Vitamin E (Trolox) addition to Tris-egg yolk extender preserves ram spermatozoon structure and kinematics after cryopreservation. Animal Reproduction Science, 137(1-2): 37-44.
Vahedi, V., Hedayat Evrigh, N., Behroozlak, M. and Dirandeh, E. (2018). Antioxidant effects of Thyme (Thymus vulgaris) extract on ram sperm quality during cryopreservation. Iranian Journal of Applied Animal Science, 8(2): 263-269.
Wang, L.G., Liu, X.M., Kreis, W. and Budman, D.R. (1997). Down-regulation of prostate-specific antigen expression by finasteride through inhibition of complex formation between androgen receptor and steroid receptor binding consensus in the promoter of the PSA gene in LNCaP cells. Cancer Research, 57(4): 714-719.
World Health Organization. (2010). WHO laboratory manual for the examination and processing
Of human semen. 5th ed., pp: 149.
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.
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Ahmed, N.A.H., Nahed, M. and El-Aal, R.A. (2015). Assessing the effect of thyme and rosmary as antiaflatoxicosis on fertility in male rats. The Journal of American Science, 11(12): 294-302.
Alasadiy, Y.D.K. (2014). Study of the biological effect of Thymus vulgaris extracts on spermatogenesis in experimentally infected white mice Balb/c by Toxoplasma gondii. International Journal of Scientific and Engineering Research, 5(10): 547-553.
Ariza-Nieto, C., Bandrick, M., Baidoo, S.K., Anil, L., Molitor, T.W. and Hathaway, M.R. (2011). Effect of dietary supplementation of oregano essential oils to sows on colostrum and milk composition, growth pattern and immune status of suckling pigs. Journal of Animal Science, 89(4): 1079-1089.
Bailey, J.L., Blodeau, J.F. and Cormier, N.Y. (2000). Semen cryopreservation in domestic animals: A damaging and capacitating phenomenon mini Journal of Andrology, 21(1): 1-7.
Bittencourt, R.F., Bicudo, S.D., Biscarde, E.A., Chalhoub, M., Filho, A.D.L. and Oba, E. (2014). Trehalose and Calcium chelator for ram semen cryopreservation. Archives of Verterinary Science, 19(2): 66-77.
Chikhoune, A., Stouvenel, L., Iguer-Ouada, M., Hazzit, M., Schmitt, A., Lores, P., et al. (2015). In-vitro effects of Thymus munbyanus essential oil and Thymol on human sperm motility and function. Reproductive Biomedicine Online, 31(9): 411-420.
Cunha, D., Brito, B., Evangelista, J., Junior, F., Pereira, L., Rebeiro, 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.
Daghigh kia, H., Sadeghi, F., Ebrahimi, M. and Samadian, F. (2017). Comparative effect of different concentrations of hydro-ethanolic extract of chamomile on freeze-thawn semen quality of rams. Journal of Veterinary Clinical Pathology, 41(11): 13-24. [In Persian]
Daghigh Kia, H., Farhadi, R., Ashrafi, I. and Mehdipour, M. (2016). Anti-oxidative effects of ethanol extract of Origanum vulgare on kinetics, microscopic and oxidative parameters of cryopreserved Holstein bull spermatozoa. Iranian Journal of Applied Animal Science, 6(4): 901-907.
Eriksson, B.M. and Rudriguez-Martinez, H. (2000). Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa frozen in Flat Packs and Maxi-Straws. Animal Reproduction Science, 63(3-4): 205-220.
Esterabauer, H. and Cheeseman K.H. (1990). Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods in Enzymology, 186(3): 407-421.
Gedicoglu, A., Sokmen, M. and Civit, A. (2019). Evaluation of Thymus vulgaris and Thymbra spicata essential oils and plant extracts for chemical composition, antioxidant and antimicrobial properties. Food Science and Nutrition, 7(5): 1704-1714.
Hamzawy, M.A., El-Denshary, E.S.M., Hassan, N.S., Manaa, F. and Abdel Wahhab, M.A. (2012). Antioxidant and renoprotective effects of Thymus vulgaris extracts in rats during aflatoxicosis. Global Journal of Pharmacy and Pharmacology, 6(2): 106-117.
Hazzit, M., Baaliouamer, A., Faleiro, M.L. and Miguel, M.G. (2006). Composition of the essential oils of Thymus and Origanum species from Algeria and their antioxidant and antimicrobial activities. Journal of Agricultural and Food Chemistry, 54(17): 6314-6321.
Hu, J.H., Chen, X.Y., Li, G., Li, Q.W., Yang, H., Wang, L.I., et al. (2006). The cryopreservation effect on frozen-thawed semen of egg yolk low density lipoproteins. Journal of Animal Science, 19(4): 486- 494.
Ieropoli, S., Do Espirito Santo, M., Masullo, P. and Sansone, G. (2004). Effect of extender composition, cooling rate and freezing on the fertilization viability of spermatozoa of the Pacific oyster (Crassostrea gigas). Cryobiology, 49(3): 250-257.
Jalili, C., Kamani, M., Roshankhah, S., Sadeghi, H. and Salahshoor, M.R. (2018). Effect of Falcaria vulgaris extracts on sperm parameters in diabetic rats. Andrologia, 50(10): 1-9.
Lupold, S. and Fitzpatrick, J.L. (2015). Sperm number trumps sperm size in mammalian ejaculate evolution. Proceedings of the Royal Society, 282(1): 1-7.
Maia, J.D., La Corte, R., Martinez, J., Ubbink, J. and Prata, A.S. (2019). Improved activity of Thyme essential oil (Thymus vulgaris) against Aedes aegypti larvae using a biodegradable controlled release system. Industrial Crops and Products, 136(2): 110-120.
Malo, C., Gil, L., Cano, R., Martinez, F. and Gale, I. (2011). Antioxidant effect of rosmary (Rosmarinus officinalis) on boar epididymal spermatozoa during cryopreservation. Theriogenology, 75(9): 1735-1741.
Masoudi, R., Shahneh, A.Z. and Asadzadeh, N. (2019). Improvement of ram’s frozen sperm quality using Glutathione antioxidant. Veterinary Research and Biological Products, 33(128): 69-75. [In Persian]
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.
Nateq, S., Moghaddam, G.H., Alijani, S. and Behnam, M. (2020). The effects of different levels of Nano selenium on the quality of frozen-thawed sperm in ram. Journal of Applied Animal Reaserch, 48(1): 434-439.
Novak, J., Bitsch, C., Langbehn, J., Pank, F., Skoula, M., Gotsiou, Y., et al. (2000). Ratios of cis- and trans-sabinene hydrate in Oryganum majorana L. and Origanum microphyllum (Bentham) Vogel. Biochemical Systematics and Ecology, 28(7): 697-704.
Ozguven, M. and Tansi, S. (1998). Drug yield and essential oil of Thymus vulgaris L. as in influenced by ecological and ontogenetical variation. Turkish Journal of Agriculture and Forestry, 22(6): 537- 542.
Salahshoor, M.R., Abdolmaleki, A., Faramarzi, A., Ziapour, A. and Roshankhah, S. (2020). Thymus vulgaris attenuates Myleran-induced reproductive damage by decreasing oxidative stress and Lipid peroxidation in male rats. Journal of Human Reproductive Sciences, 13(1): 38-45.
Shanoon, A.K. and Mahdi, S.J. (2012). Effects of Thymus vulgaris and Zingiber officinale aqueous on semen parameters, testes weight and histology measurement of broiler breeder male. International Journal of Poultry Science, 11(9): 594-598.
Shafaati, P. and Moghaddam, G.H. (2021). The effect of morphometry, viscosity and liquefaction on frozen sperm quality of Ghezel ram. Journal of Veterinary Clinical Pathology, 58(15): 155-167. [In Persian]
Shafaati, P., Moghaddam, G.H., Daghigh kia, H. and Alijani, S. (2020). Investigating the effect of diluents containing EDTA and Propylene glycol on survival of frozen semen of Ghezel ram. Journal of Veterinary Clinical Pathology, 52(13): 353-369. [In Persian]
Sharafi, M.M., Forouzanfar, S.M., Hosseini, M., Hajian, M., Ostadhosseini, S., Hosseini, L., et al. (2009). In vitro comparison of soybean lecithin based- extender with commercially available extender for ram semen cryopreservation. International Journal of Fertility and Sterility, 3(3): 149-152.
Sicherle, C.C., Maia, M.S., Bicudo, S.D., Rodello, L. and Azqvedo, H.C. (2011). Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen supplemented with catalase or Torolox. Small Ruminant Research, 95(2-3): 144-149.
Silva, S.V., Soares, A.T. Batista, A.M., Almeida, F.C., Nunes, J.F. and Peixoto, C.A. (2013). Vitamin E (Trolox) addition to Tris-egg yolk extender preserves ram spermatozoon structure and kinematics after cryopreservation. Animal Reproduction Science, 137(1-2): 37-44.
Vahedi, V., Hedayat Evrigh, N., Behroozlak, M. and Dirandeh, E. (2018). Antioxidant effects of Thyme (Thymus vulgaris) extract on ram sperm quality during cryopreservation. Iranian Journal of Applied Animal Science, 8(2): 263-269.
Wang, L.G., Liu, X.M., Kreis, W. and Budman, D.R. (1997). Down-regulation of prostate-specific antigen expression by finasteride through inhibition of complex formation between androgen receptor and steroid receptor binding consensus in the promoter of the PSA gene in LNCaP cells. Cancer Research, 57(4): 714-719.
World Health Organization. (2010). WHO laboratory manual for the examination and processing
Of human semen. 5th ed., pp: 149.
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.