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 (LR), 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, LR and Beltsville Poultry Semen Extender (BPSE). Semen was cryopreserved in 0.5 ml French straws. Thawing was done at 5°C for 100 sec in ice water in Experiments 1, 2 and 4, whereas in Experiment 3 thawing was done at 37°C for 30 sec. The post-thaw sperm motility, viable sperm and 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 LR treatment 48.12 and 30.89% respectively. In conclusion, using cryoprotectant EG (8%) and thawing at 37°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.
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