The study was conducted on in-vitro fertility assessment of frozen thawed semen collected from Kundhi buffalo bull maintained at Department of Animal Reproduction, Faculty of Animal Husbandry and Veterinary Sciences, Sindh Agriculture University Tandojam. Before freezing of semen, each ejaculate was assessed for volume, sperm concentration, mass activity and moss motility percentage. Twenty semen samples having motility 60% or above were frozen for post-thaw assessment. Frozen thawed semen was incubated at 250C for 5 hours and examined for progressive linear motility and live dead sperm count. The mean volume, mass activity, moss motility percentage sperm concentrations and pH of the semen were found to be 2.79±0.217 ml, 2.85±0.111, 71.75±2.621, 11.35±1.255 millions/ml and 5.8185±0.092 respectively of fresh semen. No significant difference was found between the parameters except pH, which was significantly different between the bulls. The mean sperm motility percentage and live dead sperm count % of Kundhi buffalo bull semen was found to be 20.46±1.62 and 6.9± 0.2% for frozen semen. A significant (P< 0.05) difference was found between the bulls for post-thaw motility percentage. It was found that at 01 hour incubation, 43.25±2.95% of sperms were motile having 11.78±0.28 % dead sperm count. It was gradually decline from 0 to 5 hours incubation, After 5 hours, all sperms were found dead. It is concluded that sperms maintaining long term motility and having less live dead sperms count were considered suitable for artificial insemination.