Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

71 COMPARISON OF CRYOPROTECTANTS ON CRYOPRESERVATION OF VENDA COCK SPERMATOZOA

M. L. Mphaphathi A B , M. B. Raito A , M. B. Makhafola A B , D. Luseba B and T. L. Nedambale A
+ Author Affiliations
- Author Affiliations

A ARC-API, Germplasm and Reproduction Biotechnologies, P/Bag x2, IRENE, 0062, Irene, Gauteng, Republic of South Africa;

B Tshwane University of Technology, Department of Animal Sciences, P/Bag x680, Pretoria, 0001, Pretoria, Republic of South Africa

Reproduction, Fertility and Development 21(1) 135-136 https://doi.org/10.1071/RDv21n1Ab71
Published: 9 December 2008

Abstract

Improving the cryopreservation technique for indigenous fowl semen may contribute to the development of cryogene banks in South Africa. The goal was to identify a cryoprotectant among dimethyl sulfoxide (DMSO), ethylene glycol (EG), and propanediol (PND) that is compatible with survival after freezing of Venda cock spermatozoa. Six Venda cocks were used for semen collection. The abdominal massaging technique was applied for semen collection from cocks. Individual ejaculates were diluted with modified Kobidil+ (mK+) extender (extender A) at ratio of 1:2 (v/v) before freezing, and equilibrated for 2 h. Semen was diluted again at a ratio of 1:1 (v/v) with mK+ plus 8% DMSO, EG, and PND (extender B) and equilibrated for 2 h at 5°C. Semen were then transferred into 0.25-mL plastic straws and placed into a programmable freezer (Planer Kryosave). The temperature of the chamber was decreased in a stepwise manner, from 5°C at a rate of 1°C min–1 until it reached the target temperature of –20°C. Finally, the straws were exposed to liquid nitrogen (LN2) vapor and plunged into LN2 (–196°C). The semen straws were stored in an LN2 tank at –196°C. After 1 week, frozen semen straws were thawed at 5°C for evaluation of spermatozoa survivability and motility rate at 0, 30, 60, and 90 min, using contrast microscopy (20× magnification). Data were analyzed by ANOVA. Spermatozoa live and motility rates were greater before freezing (Table 1) in all groups. There was no significance difference between DMSO and EG with regard to live and motility rates. However, the lowest rates of live and motility spermatozoa were recorded in the PND group. In conclusion, this study demonstrated that the cryopreservation process reduces sperm quality and propanediol was not suitable for cryopreserving Venda cock spermatozoa.


Table 1.  Comparison of three CPA on cryopreservation of Venda cock semen
T1

This study was funded by the South African National Department of Agriculture, ARC, DST-PDP (RT19000), and National Research Foundation (NRF, Grant. no. RT21 and 24000).