Effect of different concentrations of green tea on Cryopreservation of German Shepherd dog
Abstract
To investigate the Effect of adding different concentrations of green tea extract as an antioxidant on specific physical semen characteristics of German Shepherd dogs during various steps of frozen semen processing this study was carried out. A total (of 20) good Ejaculate semen samples were collected in a plastic bag from German Shepherd dogs aged 3-4 yearsIn different locations (dog breeding).To achieve the current study's main objectives, two experiments were designed through periods extended from the beginning of December 2021 until the end of May 2022. The investigations included studying the physical properties of raw Semen (ejaculate volume, semen color, consistency, motility percentage, progressive motility percentage, dead sperm percentage, sperm abnormalities percentage, and hypo-osmotic swelling test (HOST). After that, semen samples were diluted with Tris supplemented with antioxidants. The physical properties of Semen were followed after cooling and after freezing (post-thawing). In the experiment, ten ejaculates were collected. Dilution of Semen was performed with Tris supplemented with two T1, T2, and T3) were cooled slowly up to 5C and equilibrated, packed into 0.5 ml straws, and put in vapor nitrogen (-196C) for 5-10 minutes after that, stored for 48and 72 hr in liquid Nitrogen (-196C). Progressive motility percentage, dead sperm percentage, sperm abnormalities percentage, and hypo-osmotic swelling test (HOST) were evaluated after cooling and post-thawing (Submerging the straws in the water bath at 37C for 30 seconds). It can be concluded from the present study that adding green tea extract as an antioxidant to the semen extender improves the ability to freeze, especially in the treatment T3 (0.01 mg/5 ml) and the T2 (0.05 mg/5 ml). This additive can decrease the negative Effect of freezing in liquid Nitrogen compared to the control group.
Keywords:
sperm, German Shepherd dogs semen, cryopreservation, green tea
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References
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13. Goodrowe KL., Walker SL., Ryckman DP., Mastromonaco GF., Hay MA., Bateman HL., Waddell WT. 2000. Piecing together the puzzle of carnivore reproduction. Animal Reproduction Science 60-61:389-403.
14. Hewitt, D.A., Leahy, R., Sheldon, I.M. and England, G.C.M., 2001. Cryopreservation of epididymal dog sperm. Animal Reproduction Science 67, 101-111.
15. Hong, H.M.; Sim, G.Y.; Park, S.M.; Lee, E.J.; Kim, D.Y.2018. Ameliorative effect of chitosan complex on miniature pig sperm cryopreservation. J. Anim. Reprod. Biotech. 33, 337–342.
16. Hori, T., Haguida, K., Kawakami, E. and Tsutsui, T., 2004. Unilateral intrauterine insemination with prostatic fluid-sensitized frozen caudal epididymal sperm in Beagle dogs. Theriogenology 63, 1573-1583.
17. Hori, T., Ichikawa, M., Kawakami, E. and Tsutui, T., 2003. Artificial insemination of frozen epididymal sperm in Beagle dogs. The journal of Veterinary Medical Science 66, 37-41.
18. Hori, T., Matsuda, Y., Kobayashi, M., Kawakami, E. and Tsutsui, T., 2011. Comparison of fertility on intrauterine insemination between cryopreserved ejaculated and cauda epididymal sperm in dogs. The journal of Veterinary Medical Science 73, 1685-1688.
19. Hyon, S.H., 2004. A non-frozen living tissue bank for allotransplantation using green tea polyphenols. Yon med. J., 45: 1025-1034.
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23. Kutzler, M.A.2005.Semen collection in the dog. Theriogenology, 64, 747, 754 .
24. Mehdipour M, Daghigh Kia H, Najafi A, Vaseghi Dodaran H, García-Álvarez O.2016. Effect of green tea (Camellia sinensis) extract and pre-freezing equilibration time on the post-thawing quality of ram semen cryopreserved in a soybean lecithinbased extender. Cryobiology .73:297-303.
25. Michael, A.; Alexopoulos, C.; Pontiki, E.; Hadjipavlou-Litina, D.; Saratsis, P.; Boscos, C.2007. Effect of antioxidant supplementation on semen quality and reactive oxygen species of frozen-thawed canine spermatozoa. Theriogenology ., 68, 204–212.
26. Naresh, S.; Atreja, S.K. 2015.The protein tyrosine phosphorylation during in vitro capacitation and cryopreservation of mammalian spermatozoa. Cryobiology , 70, 211–216.
27. Noor ,H.S.2021.Effect of Ginseng (Panax Ginseng ) as a diet supplement on fertility in and out of breeding season in Awassi Rams in Basrah Province. Master. Thesis . Theriogenology of veterinary medicine in Basrah University.
28. Ozcan, S.S., Akin H., Bayram H., Bas M., Yildiz A., Oz-demiroglu A. (2009). Utilization of police dogs: a Turkish perspective. Policing: An International Journal of Police Strategies and Management, 32, 226–237
29. Park, B.J.; Lee, H.J.; Lee, S.L.; Rho, G.J.; Kim, S.J.; Lee, W.J.2018. Establishment of normal reference data of analysis in the fresh and cryopreserved canine spermatozoa. J. Anim. Reprod. Biotechnol. 33, 75–84.
30. Park, S.H.; Jeon, Y.; Yu, I.J. 2017.Effects of antioxidants supplementation in porcine sperm freezing on in vitro fertilization and the glutathione and reactive oxygen species level of presumptive zygotes. J. Anim. Reprod. Biotechnol32, 337–342.
31. Parr, J.R., Otto C.M. (2013). Emergency visits and occupa-tional hazards in German Shepherd police dogs (20082010). Journal of Veterinary Emergency and Critical Care, 23, 591–597.
32. Pena, A.I.; Barrio, M.; Becerra, J.J.; Quintela, L.A.; Herradon, P.G.2012.Motile sperm subpopulations in frozen-thawed dog semen: Changes after incubation in capacitating conditions and relationship with sperm survival after osmotic stress. Anim. Reprod. Sci. 133, 214–223.
33. Peña, F.J., Núñez-Martínez, I., Morán, J.M., 2006. Semen technologies in dog breeding: an update. Reproduction of Domestic Animals 41, 21-29.
34. Ponglowhapan, S., Gustavsson, B.E. and Forsberg, C.L. 2004. Influence of glucose and fructose in the extender during long term storage of chilled canine semen. Theriogenology, 62:1498-1517.
35. Quintero-Moreno, A., Rigau, T. and Rodriguez-Gil, J.E., 2004. Regression analyses and motile sperm subpopulation structure study as improving tools in boar semen quality analysis. Theriogenology, 61: 673–690.
36. Rota, A.; Frishling, A,; Vannozzi, I,; Camillo, F,; Romagnoli, S.2001. Effect of inclusion of skimmed milk in freezing extenders on the viability of canine spermatozoa after thawing. Journal of reproduction and fertilility. Suppl. v.57, p.377-381.
37. Silva, A. R.; Cardoso, R. C. S.; Silva, L O. M.2000. Congelação de sêmen canino com diferentes concentrações de gema de ovo e glycerol em diluidores à base de Tris e água de coco. Ci. Rur., v. 6, p. 1021-1025.
38. Sumayah,F.K.A.2017.Role of calcium chloride and /or vitamin Cin amelioration of sodium fluoride induced toxic effects on some physiological parameters in adult male rats . Master. Thesis . physiology of veterinary medicine in Basrah University.
39. Susilowati S, Sardjito T, Widodo OS, 2018. Effect of green tea extract supplementation in the semen extender on post-thaw sperm quality of Simmental bulls. Philipp Vet Med. 55:127–34.
40. Watson, P.F., and W.V. Holt. 2001. Cryobanking the Genetic Resource: Wildlife Conservation for the Future? Taylor and Francis, New York.
41. Wittayarat M, Ito A, Kimura T, Namula Z, Luu VV, DoLTK, Sato Y, Taniguchi M, Otoi T.2013. Effects of green tea polyphenolon the quality of canine semen after long-termstorage at 5oC. Reprod biol . 13: 251-254.
42. Wydooghe, E.; Snoeck, F.; Van Soom, A.2016. Establishment of live birth following intravaginal artificial insemination with chilled epididymal dog semen collected postmortem: Case report. In Proceedings of the 8th International Symposium on Canine and Feline Reproduction ISCFR, Paris, France, 25 June .
43. Yu I., S O N G S As E N N., G O D Ke R.A., L E I B O S.P. 2002. Differences among dogs in response of their spermatozoa to cryopreservation using various cooling and warming rates. Cryobiology, 44: 62–78.
44. Zaid ,Q.M.2015.Evaluation of local Bucks semen by traditional and Computerized Analyzer after injection of Prostaglandin F2 alpha.Master. Thesis . Theriogenology of veterinary medicine in Basrah University.
2. Al-Daraji, H.J., 2011. Effect of diluent supplementation with different levels of green tea on roosters’ semen quality during in vitro storage. Int. J. Pl. Anim. environ. Sci., 3: 51-56.
3. Bailey J.L., Bilodeau J.F., Cormier N. 2000: Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon. J. Androl., 21: 1-7.
4. Bansal AK, Bilaspuri GS, 2011. Impacts of oxidative stress and antioxidants on semen functions. Vet Med Int, Published online. Doi doi: 10.4061/2011/686137.
5. Cardoso, R.C.S; Silva, A.R.; Uchoa, D.C; Silva, L.D.M.2003. Cryopreservation of canine semen using a coconut water extender with egg yolk and three different glycerol concentrations. Theriogenology. v.59, p.743-751.
6. Chen L, Yang X, Jiao H, Zhao B.2003. Tea catechins protect against lead-induced ROS formation, mitochondrial dysfunction and calcium dysregulation in PC12 cells. Chem Res Toxicol . 16: 1155-1161.
7. Chyu, K.Y., Babbidge, S.M., Zhao, X., Dandillaya, R., Rietveld, A.G., Yano, J., Dimayuga, P., Cercek, B. and Shah, P.K., 2004. Differential effects of green-tea derived catechin on developing versus established atherosclerosis in apolipoprotein E-null mice. Circulation, 109: 2448 2453.
8. Esterhuizen, A.D.; Franken, D.R.; Lourens, J.G.; Prinsloo, E.; van Rooyen, L.H.2000. Sperm chromatin packaging as an indicator of in-vitro fertilization rates. Hum. Reprod. 15, 657–661.
9. Falah,A.A.2017.Effect of using some amino acid and Beet juice as a diluters for Najdi and Arabi and their crosses rams semen during different seasons year . Ph. Thesis . Animal Resources of veterinary medicine in Basrah University.
10. Freshman, J.L., 2002. Semen collection and evaluation. Clinical Techniques in Small Animal Practice 17 (3), 104-107.
11. Gadea J, Gumbao D, Matas C, Romar R 2005. Supplementation of the thawing media with reduced glutathione improves function and the in vitro fertilizing ability of boar spermatozoa after cryopreservation. J Androl 26:749 –56.
12. Gadea J, Gumbao D, Matas C, Romar R.2005.Supplementation of the thawing media with reduced glutathione improves function and the in vitro fertilizing ability of boar spermatozoa after cryopreservation. J Androl 26:749 –56.
13. Goodrowe KL., Walker SL., Ryckman DP., Mastromonaco GF., Hay MA., Bateman HL., Waddell WT. 2000. Piecing together the puzzle of carnivore reproduction. Animal Reproduction Science 60-61:389-403.
14. Hewitt, D.A., Leahy, R., Sheldon, I.M. and England, G.C.M., 2001. Cryopreservation of epididymal dog sperm. Animal Reproduction Science 67, 101-111.
15. Hong, H.M.; Sim, G.Y.; Park, S.M.; Lee, E.J.; Kim, D.Y.2018. Ameliorative effect of chitosan complex on miniature pig sperm cryopreservation. J. Anim. Reprod. Biotech. 33, 337–342.
16. Hori, T., Haguida, K., Kawakami, E. and Tsutsui, T., 2004. Unilateral intrauterine insemination with prostatic fluid-sensitized frozen caudal epididymal sperm in Beagle dogs. Theriogenology 63, 1573-1583.
17. Hori, T., Ichikawa, M., Kawakami, E. and Tsutui, T., 2003. Artificial insemination of frozen epididymal sperm in Beagle dogs. The journal of Veterinary Medical Science 66, 37-41.
18. Hori, T., Matsuda, Y., Kobayashi, M., Kawakami, E. and Tsutsui, T., 2011. Comparison of fertility on intrauterine insemination between cryopreserved ejaculated and cauda epididymal sperm in dogs. The journal of Veterinary Medical Science 73, 1685-1688.
19. Hyon, S.H., 2004. A non-frozen living tissue bank for allotransplantation using green tea polyphenols. Yon med. J., 45: 1025-1034.
20. Jiřina, K. and Anton, K., 2013. Effect of green tea extract on motility parameters of rabbit sperm. J. Microbiol. Biotech. Fd. Sci., 2: 1-13.
21. Johnston, S.D., Kustritz, M.V.R. and Olson, P.N.S. 2001. Semen collection, evaluation, and preservation. In: Canine and feline theriogenology (1st Edn.), W.B. Saunders, Philadelphia. pp. 287-306.
22. Khairi, G., A., A.,2018. Activation of Thawed Frozen Sperms of Iraqi Bucks and Fertilization of Goats Ova Using Different Additives and Media, Ph.D. in Veterinary Medicine / Physiology, Basrah.
23. Kutzler, M.A.2005.Semen collection in the dog. Theriogenology, 64, 747, 754 .
24. Mehdipour M, Daghigh Kia H, Najafi A, Vaseghi Dodaran H, García-Álvarez O.2016. Effect of green tea (Camellia sinensis) extract and pre-freezing equilibration time on the post-thawing quality of ram semen cryopreserved in a soybean lecithinbased extender. Cryobiology .73:297-303.
25. Michael, A.; Alexopoulos, C.; Pontiki, E.; Hadjipavlou-Litina, D.; Saratsis, P.; Boscos, C.2007. Effect of antioxidant supplementation on semen quality and reactive oxygen species of frozen-thawed canine spermatozoa. Theriogenology ., 68, 204–212.
26. Naresh, S.; Atreja, S.K. 2015.The protein tyrosine phosphorylation during in vitro capacitation and cryopreservation of mammalian spermatozoa. Cryobiology , 70, 211–216.
27. Noor ,H.S.2021.Effect of Ginseng (Panax Ginseng ) as a diet supplement on fertility in and out of breeding season in Awassi Rams in Basrah Province. Master. Thesis . Theriogenology of veterinary medicine in Basrah University.
28. Ozcan, S.S., Akin H., Bayram H., Bas M., Yildiz A., Oz-demiroglu A. (2009). Utilization of police dogs: a Turkish perspective. Policing: An International Journal of Police Strategies and Management, 32, 226–237
29. Park, B.J.; Lee, H.J.; Lee, S.L.; Rho, G.J.; Kim, S.J.; Lee, W.J.2018. Establishment of normal reference data of analysis in the fresh and cryopreserved canine spermatozoa. J. Anim. Reprod. Biotechnol. 33, 75–84.
30. Park, S.H.; Jeon, Y.; Yu, I.J. 2017.Effects of antioxidants supplementation in porcine sperm freezing on in vitro fertilization and the glutathione and reactive oxygen species level of presumptive zygotes. J. Anim. Reprod. Biotechnol32, 337–342.
31. Parr, J.R., Otto C.M. (2013). Emergency visits and occupa-tional hazards in German Shepherd police dogs (20082010). Journal of Veterinary Emergency and Critical Care, 23, 591–597.
32. Pena, A.I.; Barrio, M.; Becerra, J.J.; Quintela, L.A.; Herradon, P.G.2012.Motile sperm subpopulations in frozen-thawed dog semen: Changes after incubation in capacitating conditions and relationship with sperm survival after osmotic stress. Anim. Reprod. Sci. 133, 214–223.
33. Peña, F.J., Núñez-Martínez, I., Morán, J.M., 2006. Semen technologies in dog breeding: an update. Reproduction of Domestic Animals 41, 21-29.
34. Ponglowhapan, S., Gustavsson, B.E. and Forsberg, C.L. 2004. Influence of glucose and fructose in the extender during long term storage of chilled canine semen. Theriogenology, 62:1498-1517.
35. Quintero-Moreno, A., Rigau, T. and Rodriguez-Gil, J.E., 2004. Regression analyses and motile sperm subpopulation structure study as improving tools in boar semen quality analysis. Theriogenology, 61: 673–690.
36. Rota, A.; Frishling, A,; Vannozzi, I,; Camillo, F,; Romagnoli, S.2001. Effect of inclusion of skimmed milk in freezing extenders on the viability of canine spermatozoa after thawing. Journal of reproduction and fertilility. Suppl. v.57, p.377-381.
37. Silva, A. R.; Cardoso, R. C. S.; Silva, L O. M.2000. Congelação de sêmen canino com diferentes concentrações de gema de ovo e glycerol em diluidores à base de Tris e água de coco. Ci. Rur., v. 6, p. 1021-1025.
38. Sumayah,F.K.A.2017.Role of calcium chloride and /or vitamin Cin amelioration of sodium fluoride induced toxic effects on some physiological parameters in adult male rats . Master. Thesis . physiology of veterinary medicine in Basrah University.
39. Susilowati S, Sardjito T, Widodo OS, 2018. Effect of green tea extract supplementation in the semen extender on post-thaw sperm quality of Simmental bulls. Philipp Vet Med. 55:127–34.
40. Watson, P.F., and W.V. Holt. 2001. Cryobanking the Genetic Resource: Wildlife Conservation for the Future? Taylor and Francis, New York.
41. Wittayarat M, Ito A, Kimura T, Namula Z, Luu VV, DoLTK, Sato Y, Taniguchi M, Otoi T.2013. Effects of green tea polyphenolon the quality of canine semen after long-termstorage at 5oC. Reprod biol . 13: 251-254.
42. Wydooghe, E.; Snoeck, F.; Van Soom, A.2016. Establishment of live birth following intravaginal artificial insemination with chilled epididymal dog semen collected postmortem: Case report. In Proceedings of the 8th International Symposium on Canine and Feline Reproduction ISCFR, Paris, France, 25 June .
43. Yu I., S O N G S As E N N., G O D Ke R.A., L E I B O S.P. 2002. Differences among dogs in response of their spermatozoa to cryopreservation using various cooling and warming rates. Cryobiology, 44: 62–78.
44. Zaid ,Q.M.2015.Evaluation of local Bucks semen by traditional and Computerized Analyzer after injection of Prostaglandin F2 alpha.Master. Thesis . Theriogenology of veterinary medicine in Basrah University.
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30/04/2023
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N. y Abood and A. S. Aliawy, “Effect of different concentrations of green tea on Cryopreservation of German Shepherd dog”, Int J Indig Herb Drug, vol. 8, no. 2, pp. 16-30, Apr. 2023.
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