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The study was carried out to understand the effect of various concentrations of cholesterol loaded cyclodextrin (CLC) on cryosurvivability of buffalo spermatozoa. Twenty four ejaculates with mass motility ≥ 3+ from 4 bulls (6 from each bull) were collected as per standard procedure. Each ejaculate was split into four groups viz., Group I (control: extended with Tris-egg yolk-glycerol extender upto 80×106 sperm/ml), Group II (treated with CLC 1 mg/ml), Group III (treated with CLC 2 mg/ml) and Group IV (treated with CLC 3 mg/ml). Semen samples of Group II, III or IV were incubated with CLC at 37oC for 15 minutes for the entry of cholesterol into sperms and finally diluted upto 80×106 sperm/ml. The results revealed that post-dilution and after freezing at 0.7 and 30 days of cryopreservation motility, live and dead, acrosomal integrity and hypo-osmotic swelling test were significantly (P<0.05) higher in Group III as compared to all other groups. However, significantly (P<0.05) higher percentage of abnormal sperm was observed in control group as compared to other groups. Therefore, addition of CLC 2 mg/ml in extender during buffalo semen cryopreservation has beneficial role in improvement of semen quality.
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