Kısa Süreli Saklanan Epidydimal Anadolu Mandası Spermasına İlave Edilen Karnosik Asitin Etkisi

Abstract

Bu çalışma biberiyeden (rosmarinus officinalis) elde edilen karnosik asitin epididimal manda spermasının kısa süreli

saklamasında motilite, anormal spermatozoon oranları ve HOST/HE test oranları üzerine etkisini belirlemek amacıyla

yapılmıştır. Sulandırılan spermalar kesim sonrası alınan 36 baş (4-5 yaşlı) Anadolu Mandası testisinden elde edildi. Her 6

testisten toplanan spermalar 5 eşit parçaya bölünerek gruplar oluşturuldu ve çalışma 6 kez tekrar edildi. Gruplar 12.5, 25, 50

ve 100 μg/ml karnosik asit ve antioksidan içermeyen Tris sulandırıcısından oluşturuldu. Sperma + 4 0C’ye düşürüldü ve 0.,

24. ve 48. saatlerde spermatolojik veriler değerlendirildi. Motilite yönünden 0. saatte gruplar arasında herhangi bir fark

tespit edilmemiş 24. saat bulgularında ise kontrole göre tüm gruplardaki motilite değerleri yüksekti. Bunun yanında 48. saat

motilite değerleri ise 12.5, 25 ve 50 μg/ml lik gruplarda kontrol grubuna göre yüksekti. Bu değerlerin istatistiki açıdan

önemliydi (p<0.05). Toplam anormal spermatozoon yönünden karnosik asidin koruyucu etkisi sırasıyla 0. saatte 12.5

μg/ml’lik grupta, 24. saatte 12.5 ve 25 μg/ml grupta ve 48. saatte tüm gruplarda önemli bulundu (p<0.05). H+/Eyönünden

değerlendirildiğinde karnosik asitin 0. saatte 25 μg/ml’lik grupta 24 ve 48. saatlerde ise tüm gruplarda koruyucu

etkisinin önemli olduğu tespit edildi (p<0.05). Sonuç olarak Manda epididimal spermasının + 4 0C saklanmasında

sulandırıcıya eklenen karnosik asitin motilite, spermatozoon morfolojisi, membran bütünlüğü ve canlılığı açısından özellikle

12,5 ve 25 μg/ml lik dozlarının kontrol grubuna göre koruyucu etki gösterdiği belirlenmiştir.

Keywords

• Epididimal Sperma,Karnosik Asit,Kısa Süreli Saklama,Manda

Effect of Carnosic Acid On The Short Term Storage Anatolian Buffalo Epididymal Sperm

Abstract

This study was conducted to determine the effect of carnosic acid, extracted from rosemary (rosmarinus officinalis), to

epididymal buffalo sperm to short-term motility, abnormal spermatozoon rates and HOS test ratio. Semen samples were

obtained from 36 (4-5 year old) Anatolian buffalos after the slaughtering. Semen were collected from each 6 testes they

were divided into 5 equal parts and groups were formed and the study was repeated 6 times. The groups were composed

with the Tris extender containing carnosic acid 12.5, 25, 50 and 100 μg / ml and no-additive (control). Semen were cooled

to +4 0C and spermatological data was evaluated at 0, 24 and 48. hours. Although there was no difference between the

groups at the 0.h when evaluated in terms of motility, in 24. h motility values were higher in all groups than control.

Besides, 48.h motility values were higher in groups of 12.5, 25 and 50 μg / ml than control. these values were statistically

significant (p <0.05). In terms of total abnormal spermatozoon the protective effect of carnosic acidwas found

significantlyin 12.5 μg / ml group at 0 h, 12.5 and 25 μg / ml group at 24 h and all groups at 48 h.respectively. With regard

H+/E-, protective effect of carnosic acidwas determined in 25 μg/ml group and in all groups at 24 and 48 h. In

conclusion, it was determined that the doses of 12.5 and 25 μg / ml of carnosic acid added to extenderin +4 0C storage of

buffalo epididymal sperm showed a protective effect compared to the control group in terms of motility, spermatozoon

morphology, membrane integrity and viability.

Keywords

• Epididymal Sperm,Carnosic Acid,Short Term Storage,Buffalo

References

1. Almeida FC, Silva SV, Souza HM, Gomes WA, Lima Filho JAC, Wicke AA, Batista AM, Guerra MMP. Effects of glycerol, equilibration time and antioxidants on postthaw functional integrity of bovine spermatozoa directly obtained from epididymis. Andrologia. 2017; 49(3) 1-9.
2. Amann RP, Almquist JO. Reproductive capacity of dairy bulls VIII. Direct and indirect measurement of testicular sperm production. J. Dairy Sci. 1962; 45 774-781.
3. Bai N, He K, Roller M, Lai CS, Shao X, Pan MH, Ho CT. Flavonoids and phenolic compounds from Rosmarinus officinalis. J Agric Food Chem. 2010; 58(9) 5363-5367.
4. Arı UÇ, Öztürkler Y. Spermanın Kısa ve Uzun Süreli Saklanmasında Oksidatif Stres ve Antioksidan Kullanımı. Türkiye Klinikleri J Reprod Artif Insemin Special Topics. 2015; 1(3)16-21.
5. Barati F, Khaksary MM, Mohammadi, G. Cryopreservation of in situ cool stored buffalo (Bubalusbubalis) epididymal sperm. Iran. J. Vet. Res. 2009; 10(4) 339.
6. Chaveiro A, Cerqueira C, Silva J, Franco J, Moreira da Silva F.Evaluation of frozen thawed cauda epididymal sperms and in vitro fertilizing potential of bovine sperm collected from the cauda epididymal. Iran. J. Vet. Res. 2015; 16(2) 188-193.
7. Daghigh-Kia H, Olfati-Karaji R, Hoseinkhani A, Ashrafi I. Effect of rosemary (Rosmarinus officinalis) extracts and glutathione antioxidants on bull semen quality after cryopreservation, Spanish J. Agric. Res. 2014; 12(1) 98-105.
8. FAO.Liveanimals.http://www.fao.org/faostat/en/ #data/QA.Erişim tarihi: 31.07.2017.

9. Frankel EN, Huang S, Aeschbach R, Prior E. Antioxidant Activity of a Rosemary Extract and Its Constituents, Carnosic Acid, Carnosol, and Rosmarinic Acid, in Bulk Oil and Oil-in-Water Emulsion. J. Agric. Food Chem. 1996; 44(1) 131-135.
10. Goovaerts I, Hoflack G, Van Soom A, Dewulf J, Nichi M, de Kruif A, Bols P. Evaluation of epididymal semen quality using the Hamilton–Thorne analyser indicates variation between the two caudae epididymides of the same bull. Theriogenology. 2006; 66(2) 323- 330.
11. Gündoğan M, Avdatek F, Yeni D. Effect of extenders on motility, morphology and osmotic resistance parameters of ram sperm during liquid storage Revue Med. Vet. 2011; 162(11) 546-551.
12. Harayama H, Shibukawa T, Miyake M, Kannan Y, Kato S. Fructose stimulates shedding of cytoplasmic droplets from epididymal boar spermatozoa. Reprod Fertil Develop.1996; 8(7) 1039–1043.
13. Herold FC, de Haas K, Colenbrander B, Gerber D. Comparison of equilibration times when freezing epididymal sperm from African buffalo (Syncerus caffer) using Triladyl™ or Andromed®. Theriogenology 2006; 66(5) 1123-1130.
14. Hoseinzadeh-SaniSKA, BaratiF, Mahabady MKM. The effects of ex vivo cold-storage on cryopreservation of the goat (Caprus hircus) epididymal sperm. Iran. J. Reprod. Med. 2013; 11(9) 747-752.
15. Jones R. Sperm survival versus degradation in the mammalian epididymis: A hypothesis. Biol Reprod. 2004; 71(5) 1405-1411.
16. Kaabi M, Paz P, Alvarez M, Anel E, Boixo JC, Rouissi H, Herraez P, Anel L. Effects of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem. Theriogenology.2003; 60(7) 1249-1259.
17. Kumar A, Saxena A, Verma AK, Perumal P. Hypo-Osmotıc Swellıng Test On Buffalo (Bubalus Bubalis) Semen. Buffalo Bulletin. 2014; 33(1) 111-114.
18. Küçükkebabçı M, Aslan S. Evcil Dişi Mandaların Üreme Özellikleri. Lalahan Hay. Araşt. Enst. Derg. 2002; 42(2) 55–63.
19. Lambrechts H, van Niekerk FE, Coetzer WA, Cloete SW, van der Horst G. The effect of cryopreservation on the survivability, viability and motility of epididymal African buffalo (Syncerus caffer) sperms. Theriogenology 1999; 52(7) 1241- 1249.
20. Lee CN, Handrow RR, Lenz RW, Ax RL. Interactions of seminal plasma and glycosaminoglycans on acrosome reactions in bovine spermatozoa in vitro. Gamete research. 1985; 12(4) 345-355.
21. Lima ICS, Andrade IRA, Aguiar GV, Silva MM, Catunda AGV, Martins GA, Gadelha CRF, Campos ACN.In vitro evaluation of goat cauda epididymal sperm, cooled in different extenders at 4ºC. Arch. Zootec. 2013; 62(239) 429-437.
22. Lopez-Jimenez A, Garcıa-Caballero M, Medina MA, Quesada AR. Antiangiogenic properties of carnosol and carnosic acid, two majör dietary compounds from rosemary. Eur. J. Nutr. 2013; 52(1) 85–95.
23. Luno L, Gil M, Olaciregui N, Gonzalez RA, Jerez IB. Rosmarinic acid improves function and in vitro fertilising ability of boar sperm after cryopreservation. Cryobiology. 2014; 69(1) 157–162.
24. Malo C, Gil L, Gonzalez N, Martinez F, Cano R, de Blas I, Espinosa E. Anti-oxidant supplementation improves boar sperm characteristics and fertility after cryopreservation: Comparison between cysteine and rosemary (Rosmarinus officinalis). Cryobiology. 2010; 61(1) 142– 147.
25. Martinez-Pastor F, Guerra C, Kaabi M, Diaz AR, Anel E, Herraez P, Anel L. Decay of sperm obtained from epididymes of wild ruminants depending on postmortem time. Theriogenology. 2005; 63(1) 24-40.
26. Motlagh MK, Sharafi M, Zhandi M, Mohammadi-Sangcheshmeh A, Shakeri M, Soleimani M, Zeinoaldini S. Antioxidant effect of rosemary (Rosmarinus officinalis L.) extract in soybean lecithinbased semen extender following freeze– thawing process of ram sperm. Cryobiology. 2014; 69(2), 217-222.
27. Mulinacci N, Innocenti M, Bellumori M, Giaccherini C, Martini V, Michelozzi M.Storage method, drying processes and extraction procedures strongly affect the phenolic fraction of rosemary leaves: an HPLC/DAD/MS study. Talanta. 2011; 85(1) 167-176.
28. Muradás PR, Weiss RR, Kozicki LE, Granemann LC, Santos IW, Pimpão CT. Some viability parameters from equine spermatozoa harvested by artificial vagina and by epididymal tail washing. Arch Vet Sci. 2006; 11(3) 69-74.
29. Patrizio P, Silber S, Ord T, Balmaceda J, Asch R Two births after microsurgical sperm aspiration in congenital absence of vas deferens. The Lancet. 1988; 332(8624) 1364.
30. Songsasen N, Tong J, Leibo SP. Birth of live mice derived by in vitro fertilization with spermatozoa retrieved up to 24 hours after death. J. Exp. Zool. 1998; 280 189-196.
31. Taşdemir U, Tuncer PB, Büyükleblebici S, Özgurtas T, Durmaz E, Büyükleblebici O. Effects of Various Antioxidants on Cryopreserved Bull Sperm Quality. Kafkas Univ. Vet. Fak. Derg. 2014; 20(2) 253-258.
32. Tittarelli C, Savignone CA, Arnaudín E, Stornelli MC, Stornelli MA, De La Sota RL. Effect of storage media and storage time on survival of spermatozoa recovered from canine and feline epididymides. Theriogenology. 2006; 66(6-7) 1637-1640.
33. Tuncer PB, Çevik M, Kinet H. Holstayn Boğa Spermalarının +40C’de Saklanmasında Farklı Sperma Sulandırıcılarının Motiliteye Etkisi. Lalahan Hay. Arast. Enst. Derg. 2005; 45 (2) 1–7.
34. TÜİK.http://www.tuik.gov.tr/PreHaberBultenleri. do?id=21871. Erişim tarihi: 31.07.2017.
35. Uçar M, Gündoğan M, Yılmaz O. Mandalarda Mevsimsel Üreme Özellikleri ve Folliküler Dinamikler. Hay. Araş. Derg. 2005; 15(2) 24–29.
36. Vernet P, Aitken R, Drevet J. Antioxidant strategies in the epididymis. Molecular and cellular endocrinology. 2004; 216(1) 31-39.
37. Watson PF.Use of Giemsa stain to detect changes in the acrosome of frozen ram spermatozoa Vet Rec. 1975; 97(1) 12-15.
38. Yu I, Leibo SP. Recovery of motile, membraneintact spermatozoa from canine epididymides store for 8 days at 4°C. Theriogenology.2002; 57(3) 1179-1190.
39. Yulnawati, H, Maheshwari, H, Herdis and Rizal, M. Viability and plasma membrane integrity of the spotted buffalo epididymal spermatozoa after thawing with the addition of dextrose into the extender. Biotropia. 2009; 16(1) 21-27.
40. Zanganeh Z, Zhandi M, Zare-Shahneh A, Najafi A, Nabi MM, Mohammadi- Sangcheshmeh A. Does rosemary aqueous extract improve buck semen cryopreservation? Small Rum. Res. 2013;114 120–125.