Comparative study on effects of soy lecithin and egg yolk-based extenders on canine semen preservation at refrigeration temperature

SUMAYYA  S R; SUCHITRA  B R; RENUKARADHYA  G J; G P  KALMATH; NARASIMHA  MURTHY; MOHAN  H V

doi:10.56093/ijans.v94i12.153123

Authors

SUMAYYA S R Veterinary College, Hebbal, Bengaluru (KVAFSU), Bidar, Karnataka 560 024 India
SUCHITRA B R Veterinary College, Hebbal, Bengaluru (KVAFSU), Bidar, Karnataka 560 024 India
RENUKARADHYA G J Veterinary College, Hebbal, Bengaluru (KVAFSU), Bidar, Karnataka 560 024 India
G P KALMATH Veterinary College, Hebbal, Bengaluru (KVAFSU), Bidar, Karnataka 560 024 India
NARASIMHA MURTHY Veterinary College, Hebbal, Bengaluru (KVAFSU), Bidar, Karnataka 560 024 India
MOHAN H V Veterinary College, Hebbal, Bengaluru (KVAFSU), Bidar, Karnataka 560 024 India

https://doi.org/10.56093/ijans.v94i12.153123

Keywords:

Canine semen, Chilling, Semen extenders, Seminal attributes

Abstract

The objective of the study was to evaluate the effect of addition of Tris soy lecithin (TSL) and Tris egg yolk- based (TEY) extenders on canine seminal attributes during 7 days storage at 5°C. Semen was manually collected from fifteen apparently healthy male dogs, which then underwent semen evaluation. Only semen sample with spermatozoa motility >80% and spermatozoa concentration >200×10⁶ spermatozoa/mL were used in the present study. After initial evaluation, sperm-rich fractions were divided into two equal parts and extended with 20% TEY and 1% TSL extenders at 1:4 dilution, then stored at a refrigeration temperature of 5°C for 7 days. Chilled semen samples were assessed for pH, motility, viability, abnormality, and plasma membrane integrity for a week on daily basis. DNA integrity was assessed on days 0, 1, 3, and 5 of storage. Significant differences were observed in all seminal attributes except DNA integrity between fresh and extended semen. The pH of the semen did not differ significantly between the extenders up to day 6, indicating almost equal buffering capacity of the extenders. A significant difference in sperm motility, viability, morphological abnormality, and plasma membrane integrity was observed between the extenders from storage days 4, 3, 1, and 5, respectively. No significant difference in sperm DNA integrity was observed between the extenders. In conclusion, while both extenders were equally effective in preserving sperm DNA integrity, Tris egg yolk extender outperformed Tris soy lecithin extender in all other seminal attributes assessed in this study.

Downloads

Download data is not yet available.

References

Aboagla E M E and Terada T. 2004. Effects of egg yolk during the freezing step of cryopreservation on the viability of goat spermatozoa. Theriogenology 62(6): 1160–72.

Agarwal A, Gupta S and Sharma R. 2016. Eosin-nigrosin staining procedure, pp. 73-77. Andrological Evaluation of Male Infertility: A Laboratory Guide, 1st edn. Springer, USA.

Astuti S. 2012. Isoflavon kedelai dan potensinya sebagai penangkap radikal bebas. Jurnal Teknologi & Industri Hasil Pertanian 13(2): 126–36.

Axner E and Lagerson E. 2016. Cryopreservation of dog semen in a tris extender with 1% or 2% soya bean lecithin as a replacement of egg yolk. Reproduction in Domestic Animals 51(2): 262–68.

Batista M, Santana M, Alamo D, Gonzalez F, Nino T, Cabrera F and Gracia A. 2012. Effects of incubation temperature and semen pooling on the viability of fresh, chilled and freeze‐ thawed canine semen samples. Reproduction in Domestic Animals 47(6): 1049–55.

Beccaglia M, Anastasi P and Luvoni G C. 2009. Freezing of canine semen in an animal free protein extender. Veterinary research communications 33: 77–80.

Dalmazzo A, Losano J D A, Rocha C C, Tsunoda R H, Angrimani D D S R, Mendes C M, Assumpcao M E O D A, Nichi M and Barnabe V H. 2018. Effects of soy lecithin extender on dog sperm cryopreservation. Journal of Animal Science and Biotechnology 29(3): 174–82.

Forouzanfar M, Sharafi M, Hosseini S M, Ostadhosseini S, Hajian M,Hosseini L, Abedi P, Nili N, Rahmani H R and Nasresfahani M H. 2010. In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. Theriogenology 1(73): 480–87.

Fukui Y, Kohno H, Togari T, Hiwasa M and Okabe K. 2008. Fertility after artificial insemination using a soybean-based semen extender in sheep. Journal of Reproduction and Development 54(4): 286–89.

Hermansson U, Johannisson A and Axner E. 2021. Cryopreservation of dog semen in a Tris extender with two different 1% soybean preparations compared with a Tris egg yolk extender. Veterinary Medicine and Science 7(3): 812–19.

Hori T, Yoshikuni R, Kobayashi M and Kawakami E. 2014. Effects of storage temperature and semen extender on stored canine semen. The Journal of Veterinary Medical Science 76(2): 259–63.Johnston S D, Root Kustritz M V and Olson P N S. 2001. Semen collection, evaluation, and preservation, pp.287-306. Canine and Feline Theriogenology. 1st edn, W. B. Saunders Company, Philadelphia.

Kasimanickam V R, Kasimanickam R K, Memon M A and Rogers H A. 2012. Effect of extenders on sperm mitochondrial membrane, plasma membrane and sperm kinetics during liquid storage of canine semen at 50C. Animal Reproduction Science 36(1): 139–45.

Kmenta I, Strohmayer C, Muller-Schlosser F and Schafer Somi S. 2011. Effects of a lecithin and catalase containing semen extender and a second dilution with different enhancing buffers on the quality of cold-stored canine spermatozoa. Theriogenology 75(6): 1095–03.

Linde-Forsberg C. 1991. Achieving canine pregnancy by using frozen or chilled extended semen. Veterinary Clinics of North America: Small Animal Practice 21(3): 467–85.

Lopes G, Simoes A, Ferreira P, Martins-Bessa A and Rocha A. 2009. Differences in preservation of canine chilled semen using different transport containers. Animal Reproduction Science 112(2): 158–63.

Manjunath P, Nauc V, Bergeron A and Menard M. 2002. Major proteins of bovine seminal plasma bind to the low-density lipoprotein fraction of hen’s egg yolk. 2000. Biology of Reproduction 67(4): 1250–58.

Mingkun Sun, Wangcheng Song, ChangrongLv, Yu Li and Song H. 2022. Soybean lecithin-based extender improves the quality of chilled canine sperm. Journal of Veterinary Science and Animal Husbandry 10(1): 102–13.

Ponglowhapan S, Essen-Gustavsson B and Forsberg C L. 2004. Influence of glucose and fructose in the extender during long- term storage of chilled canine semen. Theriogenology 62(8): 1498–1517.

Pradana M Y W, Bebas W and Puja K. 2016. The effect of soyabean extender on viability and DNA integrity of Kintamani dog sperm on cold storage. Veterinary Science and Medicine Journal 4(1): 11–16.

Puja I K, Sawitri N M, Maharani N, Gunawan I W N F and Heryani L G S S. 2018. A comparative study on the effects of coconut water-based extenders on the quality of Kintamani dog semen preserved at 40C. Advances in Animal and Veterinary Sciences 6(5): 192–96.

Ramu S and Jeyendran R S. 2013. The hypo-osmotic swelling test for evaluation of sperm membrane integrity. Methods in Molecular Biology 927: 21–25.

Ray K, Jha A K, Biswas P and Basu S. 2019. Study on physicomorphological seminal characters of German shepherd canine breed. Indian Journal of Animal Health 58(1): 95–100. Rota A, Strom B and Linde-Forsberg C. 1995. Effects of seminal plasma and three extenders on canine semen stored at 40C.

Theriogenology 44(6): 885–900.

Saraswat S, Kharche S D and Jindal S K. 2014. Impact of reactive oxygen species on spermatozoa: A balancing act between beneficial and detrimental effects. Iranian Journal of Applied Animal Science 4(2): 247–55.

Shamsi M B, Imam S N and Dada R. 2011. Sperm DNA integrity assays: diagnostic and prognostic challenges and implications in the management of infertility. Journal of Assisted Reproduction and Genetics 28(11): 1073–85.

Sousa P C, Santos E A, Souza A L, Lima, G L, Barros F F, Oliveira M F and Silva A R. 2013. Sperm morphological and morphometric evaluation in captive collared peccaries (Pecari tajacu). Pesquisa Veterinaria Brasileira 33: 924–30.

Srinivas R T, Reddy K C S, Venkata Ramana K and Nagaraj P. 2022. Studies on extension and preservation of canine semen by addition of catalase at refrigeration temperature. Journal of pharmaceutical innovation 11(6): 655–58.

Ubah S A, Sule M, Chibuogwu I C, Columbus P K, Abah K O, Agbonu O A, Ejiofor C E, Mshelbwala P P and Bankole S A. 2019. Comparative study of chicken egg yolk and quail egg yolk in two chilled canine semen extenders. Sokoto Journal of Veterinary Sciences 17(4): 62–69.

Verstegen J P, Onclin K and Iguer-Ouada M. 2005. Long-term motility and fertility conservation of chilled canine semen using egg yolk added Tris–glucose extender: In vitro and in vivo studies. Theriogenology 64(3): 720–33.