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Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi

Year 2021, Volume: 14 Issue: 4, 492 - 498, 31.12.2021
https://doi.org/10.30607/kvj.1011612

Abstract

Sunulan çalışmanın amacı, dondurulmuş çözdürülmüş Malaklı Çoban Köpeği spermasında karnosik asitin motilite, morfoloji, canlılık ve membran bütünlüğü parametrelerine etkisini belirlemektir. Bu çalışmada beş baş Malaklı köpeğine ait ejakülatların spermatozoon bakımından zengin kısmı kullanıldı. Ejakülatlar birleştirildi ve beş eşit gruba bölündü. Kontrol grubu temel Tris sulandırıcısıyla sulandırıldı ve diğer gruplar 10 µM, 50 µM, 100 µM, 200 µM karnosik asit içeren gruplar oluşturuldu. Bir buçuk saat ekilibrasyonun ardından, payetler azot buharında donduruldu ve daha sonra sıvı azot içinde depolandı. Ardından dondurulmuş payetler 37oC sıcak suda çözdürüldü ve motilite, morfoloji, canlılık ve membran bütünlüğü parametreleri değerlendirildi. En yüksek motilite ve HOST-Eozin değerleri 10 µM’lık grupta elde edildi ve sırasıyla %65,00±2,23 ve %69,83±0,70 olarak belirlendi, en düşük anormal spermatozoon oranı ise 9,50±1,52 ile yine10 µM’lık gruptan elde edildi ve bu gruplardaki fark (p<0.05) kontrol grubuna göre istatistiki olarak önemli bulundu. Sonuç olarak Malaklı köpeği spermasının dondurulmasında 10 µM karnosik asitin, motilite, spermatozoon morfolojisi, membran bütünlüğü ve canlılığı açısından kontrol grubuna kıyasla koruyucu bir etki gösterdiği belirlenmiştir.

Supporting Institution

Afyon Kocatepe Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

15.SAĞ.BĠL.03

References

  • Agarwal A, Duraırajanayagam D, Halabı J, Peng J, Vazquez-Levın M. Proteomics, oxidative stress and male infertility. Reproductive BioMedicine Online, 2014; 29: 32– 58.
  • Aitken RJ. Pathophysiology of human spermatozoa. Curr Opin Obstet Gynecol, 1994; 6: 128-35.
  • Akçay E. Kangal Çoban köpeklerinde androlojik muayeneler ve spermanın dondurulması. Doktora Tezi. Ankara Üniversitesi Sağlık Bilimleri Enstitüsü, Ankara, 2000.
  • Akkuş İ. Serbest radikaller ve fizyopatolojik etkileri. Mimoza Yayınları, Konya, 1995; 32.
  • Alvarez JG, Storey BT. Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa. Mol Reprod Dev, 1995; 42: 334–346.
  • Atasoy F. Köpek-Kedi Yetiştiriciliği Ders Notları, Ankara Üniversitesi Veteriner Fakültesi, Zootekni Anabilim Dalı, Ankara,2010. Biochemistry and Biophysic, 453, 224–236.
  • Borghei-Rad SM, Zeinoaldini S, Zhandi M, Moravej H, Ansari M. Feeding rosemary leaves powder ameliorates rooster age-related subfertility. Theriogenology, 2017; 101: 35–43.
  • Breque C, Surai PF, Brillard JP. Roles of antioxidants on prolonged storage of avian spermatozoain vivoandin vitro. Mol. Reprod. Dev. 2003; 66: 314–323.
  • Bucak MN, Tekin N. Kriyoprotektanlar ve gamet hücrelerinin dondurulmasında kriyoprotektif etki. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 2007; 54: 67-72.
  • Bucak MN, Tuncer PB, Sarıözkan S, Başpınar N, Taşpınar M, Çoyan K, Bilgili A, Akalın PP, Büyükleblebici S, Aydos S, Ilgaz S. Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: antioxidants protect DNA integrity against cryodamage. Cryobiology, 2010; 61: 248–53.
  • Cupps PT. Reproduction in the dog and cat, Reproduction in Romestic animals, Editör: Cupps, P. T. California: Academic Press. 1991.
  • Cuvelier ME, Richard H, Berset C. Antioxidative activity and phenolic composition of pilot-plant and commercial extracts of sage and rosemary. J. Am. Oil. Chem. Soc., 1996; 73: 645–652.
  • 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.
  • Demirci E. Evcil hayvanlarda reprodüksiyon, suni tohumlama ve androloji ders notları (1nd ed). F.Ü.Vet.Fak., Ders Teksiri No:53, Elazığ, 2002.
  • Ducci M, Gaazano A, Villani C, Cela V, Artini PG, Martelli F, Genazzani AR. Membrane integrity evaluation in rabbit spermatozoa. Eur. J. Obstet. Gynaecol. Reprod. Biol., 2002; 102: 53–56.
  • Frankel EN, Huang SW, 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. Journal of Agricultural and Food Chemistry, 1996; 44: 131-135.
  • Griveau JF, Le Lannou D. Reactive oxygen species and human spermatozoa: physiology and pathology. Int. J. Androl. 1997; 20: 61–69.
  • Gündoğan M, Avdatek F, Yeni D. Effect of extenders on motility, morphology and osmotic resistance parameters of ram sperm during liquid storage. Revue Méd. Vét., 2011; 162 (11): 546-551.
  • Heidari-Vala H, Hariry RE, Sadeghi MR, Akhondi MM, Novin MG, Heidari M. Evaluation of an aqueous-ethanolic extract from Rosmarinus officinalis (Rosemary) for its activity on the hormonal and cellular function of testes in adult male rat. Iranian journal of pharmaceutical research: IJPR, 2013; 12(2): 445.
  • Holt WT. Fundemental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology, 2000; 53: 47-58.
  • İnanç ME, Tekin K, Akkurt MY, Olgac KT, Yılmaz B, Çil B, Kızılaslan M, Taşdemir U, Tuncer PB,Büyükleblebici S, Uysal O, Çınar Kul B. Genomewide association of male reproductive traits in Aksaray Malakli dogs. Reprod Dom Anim., 2018; 53:1555– 1562.
  • İnanç ME, Tekin K, Olgac, KT, Yılmaz B, Çil B, Taşdemir U, Tuncer PB, Büyükleblebici S, Uysal O. Effect of cholesterol loaded cyclodextrin on semen cryopreservation of Aksaray Malakli shepherd dogs of different ages. Animal Reproduction Science, 2018; 193, 191-200.
  • Kim JG, Parthasarathy S. Oxidation and the spermatozoa. Semin Reprod Endocrinol, 1998; 16(4): 235-9.
  • Leibo SP, Brandley L. Comparative cryobiology of mammalian spermatozoa, The Male Gamet: From Basic Science to Clinical Applications. Editör: Gagnon, C. St Louis: Cache River Press,1999.
  • Lemma A. Effect of Cryopreservation on Sperm Quality and Fertility, Artificial Insemination in Farm Animals. Editör: Manafi, M. InTech, 2011.
  • Malo C, Gil L, Gonzalez N, Martinez F, Cano R, De Blas I, Espınosa 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.
  • 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.
  • Özbeyaz C. “Kangal köpeklerinde bazı morfolojik özellikler”, Lalahan Hay. AraĢ. Ens. Derg., 1994; 34(1-2): 38-46.
  • Özgüneş AF, Çiftçi N. “Kangal köpeği hakkında genel bilgiler”, Tigem Derg., 1993; 1-8.
  • Pérez-Fons L, Aranda FJ, Guillén J, Villalaín J, Micol V. Rosemary (Rosmarinus officinalis) diterpenes affect lipid polymorphism and fluidity in phospholipid membranes. Archives of biochemistry and biophysics, 2006; 453(2), 224-236.
  • Rangan U, Bulkley GB. Prospects for treatment of free radical-mediated tissue injury. Br Med Bull, 1993; 49(3): 700-18.
  • Rašković A, Milanović I, Pavlović N, Ćebović T, Vukmirović S, Mikov M. Antioxidant activity of rosemary (Rosmarinus officinalisL.) essential oil and its hepatoprotective potential. BMC Complement. Altern. Med., 2014; 14: 225.
  • Rıchheimer SL, Matthew WB, Greg AK, Michael CK, David TB. Antioxidant activity of lipid-soluble phenolic diterpenes from rosemary. J. Am. Oil Chem. Soc., 1996; 73: 507–514.
  • Surai P, Cerolini S, Wisharta GJ, Speake BK, Noble RC, Sparks NHC. Lipid and antioxidant composition of chicken semen and its susceptibility to peroxidation. Avian. Poult. Biol. Rev., 1998; 9(l):11–23.
  • Surai P, Fisinin VI. Selenium in poultry breeder nutrition:An update. Anim. Feed. Sci. Technol., 2014; 191: 1–15.
  • Surai P. Antioxidant systems in poultry biology:Superoxide dismutase. Anim. Nutr., 2016; 1: 1–17.
  • Tekin N. Spermanın Muayenesi ve Değerlendirilmesi. In: Alaçam, E. Ed. Evcil Hayvanlarda Reprodüksiyon Suni Tohumlama, Doğum ve Ġnfertilite. Dizgievi, Konya, 1994; 69-79.
  • Touazi L, Aberkane B, Bellik Y, Moula N, Iguer-Ouada M. Effect of the essential oil of Rosmarinus officinalis (L.) on rooster sperm motility during 4°C short-term storage, Veterinary World, 2018; 11(5): 590-597.
  • Türk G, Çeribaşı AO, Şimşek ÜG, Çeribaşı S, Güvenç M, Kaya ŞÖ, Çiftçi M, Sönmez M, Yüce A, Bayrakdar A, Yaman M, Tonbak F. Dietary rosemary oil alleviates heat stress-induced structural and functional damage through lipid peroxidation in the testes of growing Japanese quail. Animal reproduction science, 2016; 164: 133-143.
  • Watson PF. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci, 2000; 60: 481-92.
  • Watson PF. Use of Giemsa stain to detect changes in the acrosome of frozen ram spermatozoa Vet Rec., 1975; 97(1): 12-15.
  • Yeni D, Avdatek F. Kısa Süreli Saklanan Epidydimal Anadolu Mandası Spermasına İlave Edilen Karnosik Asitin Etkisi. Kocatepe Vet J., 2017; 10(3): 187-195.
  • Yeni D, İnanç ME, Avdatek F, Tuncer PB, Türkmen R, Taşdemir U. Supplementation of Rosmarinic Acid Has Reduced Oxidative Stress on Bull Spermatozoa Following the Freeze Thawing Process. CryoLetters, 2018; 39(2): 156-165.
  • Zanganeh Z, Zhandi M, Zare-Shahneh A, Najafi A, Nabi MM, Mohammadi-Sangcheshmeh A. Does rosemary aqueous extract improve buck semen cryopreservation? Small Rumin. Res., 2013; 114: 120–125.

The Effect of Carnosic Acid on Semen Freezability in Malaklı Shepherd Dogs

Year 2021, Volume: 14 Issue: 4, 492 - 498, 31.12.2021
https://doi.org/10.30607/kvj.1011612

Abstract

The objective of the study was to determine the effect of carnosic acid, to Malakli Shepherd dog semen motility, morphology, viability and membran integrity parameters after frozen-thawed prosess. Five animal were used in this study and the sperm rich portion of the ejaculates were mixed and divided in to five equal groups. Each group extended with either tris as a control group and containing carnosic acid 10 µM, µM, 100 µM, 200 µM. Following equilibration for one and a half hour, the straws were frozen in nitrogen vapor and then stored in liquid nitrogen. Later, the frozen straws were thawed in a water bath for motility, morphology, viability and membran integrity parameters examination. The highest motilities and HOST-Eosin test results were detected in 10 µM group at 65.00±2.23 % and 69.83±0.70 % respectively, the lowest abnormal sperm rate were detected in 10 µM group at 9.50±1.52 % and the differences in these groups (p <0.05) were statistically significant compared to the control group. With regard conclusion, it was determined that the doses of 10 µM of carnosic acid added to extender in freezing of Malakli dog sperm showed a protective effect compared to the control group in terms of motility, spermatozoon morphology, membrane integrity and viability.

Project Number

15.SAĞ.BĠL.03

References

  • Agarwal A, Duraırajanayagam D, Halabı J, Peng J, Vazquez-Levın M. Proteomics, oxidative stress and male infertility. Reproductive BioMedicine Online, 2014; 29: 32– 58.
  • Aitken RJ. Pathophysiology of human spermatozoa. Curr Opin Obstet Gynecol, 1994; 6: 128-35.
  • Akçay E. Kangal Çoban köpeklerinde androlojik muayeneler ve spermanın dondurulması. Doktora Tezi. Ankara Üniversitesi Sağlık Bilimleri Enstitüsü, Ankara, 2000.
  • Akkuş İ. Serbest radikaller ve fizyopatolojik etkileri. Mimoza Yayınları, Konya, 1995; 32.
  • Alvarez JG, Storey BT. Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa. Mol Reprod Dev, 1995; 42: 334–346.
  • Atasoy F. Köpek-Kedi Yetiştiriciliği Ders Notları, Ankara Üniversitesi Veteriner Fakültesi, Zootekni Anabilim Dalı, Ankara,2010. Biochemistry and Biophysic, 453, 224–236.
  • Borghei-Rad SM, Zeinoaldini S, Zhandi M, Moravej H, Ansari M. Feeding rosemary leaves powder ameliorates rooster age-related subfertility. Theriogenology, 2017; 101: 35–43.
  • Breque C, Surai PF, Brillard JP. Roles of antioxidants on prolonged storage of avian spermatozoain vivoandin vitro. Mol. Reprod. Dev. 2003; 66: 314–323.
  • Bucak MN, Tekin N. Kriyoprotektanlar ve gamet hücrelerinin dondurulmasında kriyoprotektif etki. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 2007; 54: 67-72.
  • Bucak MN, Tuncer PB, Sarıözkan S, Başpınar N, Taşpınar M, Çoyan K, Bilgili A, Akalın PP, Büyükleblebici S, Aydos S, Ilgaz S. Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: antioxidants protect DNA integrity against cryodamage. Cryobiology, 2010; 61: 248–53.
  • Cupps PT. Reproduction in the dog and cat, Reproduction in Romestic animals, Editör: Cupps, P. T. California: Academic Press. 1991.
  • Cuvelier ME, Richard H, Berset C. Antioxidative activity and phenolic composition of pilot-plant and commercial extracts of sage and rosemary. J. Am. Oil. Chem. Soc., 1996; 73: 645–652.
  • 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.
  • Demirci E. Evcil hayvanlarda reprodüksiyon, suni tohumlama ve androloji ders notları (1nd ed). F.Ü.Vet.Fak., Ders Teksiri No:53, Elazığ, 2002.
  • Ducci M, Gaazano A, Villani C, Cela V, Artini PG, Martelli F, Genazzani AR. Membrane integrity evaluation in rabbit spermatozoa. Eur. J. Obstet. Gynaecol. Reprod. Biol., 2002; 102: 53–56.
  • Frankel EN, Huang SW, 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. Journal of Agricultural and Food Chemistry, 1996; 44: 131-135.
  • Griveau JF, Le Lannou D. Reactive oxygen species and human spermatozoa: physiology and pathology. Int. J. Androl. 1997; 20: 61–69.
  • Gündoğan M, Avdatek F, Yeni D. Effect of extenders on motility, morphology and osmotic resistance parameters of ram sperm during liquid storage. Revue Méd. Vét., 2011; 162 (11): 546-551.
  • Heidari-Vala H, Hariry RE, Sadeghi MR, Akhondi MM, Novin MG, Heidari M. Evaluation of an aqueous-ethanolic extract from Rosmarinus officinalis (Rosemary) for its activity on the hormonal and cellular function of testes in adult male rat. Iranian journal of pharmaceutical research: IJPR, 2013; 12(2): 445.
  • Holt WT. Fundemental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology, 2000; 53: 47-58.
  • İnanç ME, Tekin K, Akkurt MY, Olgac KT, Yılmaz B, Çil B, Kızılaslan M, Taşdemir U, Tuncer PB,Büyükleblebici S, Uysal O, Çınar Kul B. Genomewide association of male reproductive traits in Aksaray Malakli dogs. Reprod Dom Anim., 2018; 53:1555– 1562.
  • İnanç ME, Tekin K, Olgac, KT, Yılmaz B, Çil B, Taşdemir U, Tuncer PB, Büyükleblebici S, Uysal O. Effect of cholesterol loaded cyclodextrin on semen cryopreservation of Aksaray Malakli shepherd dogs of different ages. Animal Reproduction Science, 2018; 193, 191-200.
  • Kim JG, Parthasarathy S. Oxidation and the spermatozoa. Semin Reprod Endocrinol, 1998; 16(4): 235-9.
  • Leibo SP, Brandley L. Comparative cryobiology of mammalian spermatozoa, The Male Gamet: From Basic Science to Clinical Applications. Editör: Gagnon, C. St Louis: Cache River Press,1999.
  • Lemma A. Effect of Cryopreservation on Sperm Quality and Fertility, Artificial Insemination in Farm Animals. Editör: Manafi, M. InTech, 2011.
  • Malo C, Gil L, Gonzalez N, Martinez F, Cano R, De Blas I, Espınosa 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.
  • 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.
  • Özbeyaz C. “Kangal köpeklerinde bazı morfolojik özellikler”, Lalahan Hay. AraĢ. Ens. Derg., 1994; 34(1-2): 38-46.
  • Özgüneş AF, Çiftçi N. “Kangal köpeği hakkında genel bilgiler”, Tigem Derg., 1993; 1-8.
  • Pérez-Fons L, Aranda FJ, Guillén J, Villalaín J, Micol V. Rosemary (Rosmarinus officinalis) diterpenes affect lipid polymorphism and fluidity in phospholipid membranes. Archives of biochemistry and biophysics, 2006; 453(2), 224-236.
  • Rangan U, Bulkley GB. Prospects for treatment of free radical-mediated tissue injury. Br Med Bull, 1993; 49(3): 700-18.
  • Rašković A, Milanović I, Pavlović N, Ćebović T, Vukmirović S, Mikov M. Antioxidant activity of rosemary (Rosmarinus officinalisL.) essential oil and its hepatoprotective potential. BMC Complement. Altern. Med., 2014; 14: 225.
  • Rıchheimer SL, Matthew WB, Greg AK, Michael CK, David TB. Antioxidant activity of lipid-soluble phenolic diterpenes from rosemary. J. Am. Oil Chem. Soc., 1996; 73: 507–514.
  • Surai P, Cerolini S, Wisharta GJ, Speake BK, Noble RC, Sparks NHC. Lipid and antioxidant composition of chicken semen and its susceptibility to peroxidation. Avian. Poult. Biol. Rev., 1998; 9(l):11–23.
  • Surai P, Fisinin VI. Selenium in poultry breeder nutrition:An update. Anim. Feed. Sci. Technol., 2014; 191: 1–15.
  • Surai P. Antioxidant systems in poultry biology:Superoxide dismutase. Anim. Nutr., 2016; 1: 1–17.
  • Tekin N. Spermanın Muayenesi ve Değerlendirilmesi. In: Alaçam, E. Ed. Evcil Hayvanlarda Reprodüksiyon Suni Tohumlama, Doğum ve Ġnfertilite. Dizgievi, Konya, 1994; 69-79.
  • Touazi L, Aberkane B, Bellik Y, Moula N, Iguer-Ouada M. Effect of the essential oil of Rosmarinus officinalis (L.) on rooster sperm motility during 4°C short-term storage, Veterinary World, 2018; 11(5): 590-597.
  • Türk G, Çeribaşı AO, Şimşek ÜG, Çeribaşı S, Güvenç M, Kaya ŞÖ, Çiftçi M, Sönmez M, Yüce A, Bayrakdar A, Yaman M, Tonbak F. Dietary rosemary oil alleviates heat stress-induced structural and functional damage through lipid peroxidation in the testes of growing Japanese quail. Animal reproduction science, 2016; 164: 133-143.
  • Watson PF. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci, 2000; 60: 481-92.
  • Watson PF. Use of Giemsa stain to detect changes in the acrosome of frozen ram spermatozoa Vet Rec., 1975; 97(1): 12-15.
  • Yeni D, Avdatek F. Kısa Süreli Saklanan Epidydimal Anadolu Mandası Spermasına İlave Edilen Karnosik Asitin Etkisi. Kocatepe Vet J., 2017; 10(3): 187-195.
  • Yeni D, İnanç ME, Avdatek F, Tuncer PB, Türkmen R, Taşdemir U. Supplementation of Rosmarinic Acid Has Reduced Oxidative Stress on Bull Spermatozoa Following the Freeze Thawing Process. CryoLetters, 2018; 39(2): 156-165.
  • Zanganeh Z, Zhandi M, Zare-Shahneh A, Najafi A, Nabi MM, Mohammadi-Sangcheshmeh A. Does rosemary aqueous extract improve buck semen cryopreservation? Small Rumin. Res., 2013; 114: 120–125.
There are 44 citations in total.

Details

Primary Language Turkish
Subjects Veterinary Surgery
Journal Section RESEARCH ARTICLE
Authors

Hulusi Şahin This is me 0000-0003-1376-2437

Deniz Yeni 0000-0002-9105-5677

Fatih Avdatek 0000-0003-2345-8826

Project Number 15.SAĞ.BĠL.03
Publication Date December 31, 2021
Acceptance Date December 6, 2021
Published in Issue Year 2021 Volume: 14 Issue: 4

Cite

APA Şahin, H., Yeni, D., & Avdatek, F. (2021). Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi. Kocatepe Veterinary Journal, 14(4), 492-498. https://doi.org/10.30607/kvj.1011612
AMA Şahin H, Yeni D, Avdatek F. Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi. kvj. December 2021;14(4):492-498. doi:10.30607/kvj.1011612
Chicago Şahin, Hulusi, Deniz Yeni, and Fatih Avdatek. “Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi”. Kocatepe Veterinary Journal 14, no. 4 (December 2021): 492-98. https://doi.org/10.30607/kvj.1011612.
EndNote Şahin H, Yeni D, Avdatek F (December 1, 2021) Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi. Kocatepe Veterinary Journal 14 4 492–498.
IEEE H. Şahin, D. Yeni, and F. Avdatek, “Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi”, kvj, vol. 14, no. 4, pp. 492–498, 2021, doi: 10.30607/kvj.1011612.
ISNAD Şahin, Hulusi et al. “Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi”. Kocatepe Veterinary Journal 14/4 (December 2021), 492-498. https://doi.org/10.30607/kvj.1011612.
JAMA Şahin H, Yeni D, Avdatek F. Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi. kvj. 2021;14:492–498.
MLA Şahin, Hulusi et al. “Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi”. Kocatepe Veterinary Journal, vol. 14, no. 4, 2021, pp. 492-8, doi:10.30607/kvj.1011612.
Vancouver Şahin H, Yeni D, Avdatek F. Malaklı Köpeği Spermasının Dondurulabilirliği Üzerine Karnosik Asitin Etkisi. kvj. 2021;14(4):492-8.

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