Cryopreservation of Ghagus chicken semen: effect of cryoprotectants diluents and thawing temperature

Abstract

The present study evaluated the effects of cryoprotectants, semen diluents and thawing temperature during Ghagus chicken semen cryopreservation. Four different experiments were conducted; Experiment 1- semen was cryopreserved using 6% dimethylacetamide (DMA) and 2% dimethylsulfoxide (DMSO) in Sasaki diluent (SD) and Lake and Ravie diluent (LRD), Experiment 2 and 3- semen was cryopreserved using 8% Ethylene Glycol (EG) in SD, LRD and Red Fowl Extender (RFE), Experiment 4- semen was cryopreserved using 6% dimethylformamide (DMF) in SD, LRD and Beltsville Poultry Semen Extender (BPSE). Semen was cryopreserved in 0.5 ml French straws. Thawing was done at 5{degrees}C for 100 sec in ice water in Experiments 1, 2 and 4, whereas in Experiment 3 thawing was done at 37{degrees}C for 30 sec. The post-thaw sperm motility, live sperm and percent acrosome intact sperm were significantly (P<0.05) lower in cryopreserved samples in all the experiments. No fertile eggs were obtained from cryopreserved samples in Experiments 1 and 2, except for 8% EG RFE treatment where the fertility was 0.83%. In Experiments 3 and 4, highest fertility was obtained in LRD treatment 48.12 and 30.89% respectively. In conclusion, using cryoprotectant EG (8%) and thawing at 37{degrees}C for 30 sec, and DMF (6%) resulted in acceptable level of fertility in Ghagus chicken. Though the diluents influenced post-thaw in vitro semen parameters the fertility was not affected. In addition, results indicated that thawing temperature may be a critical stage in the cryopreservation protocol.