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Vaginal cytology is a simple technique that can be used to determine the stages of the estrus cycle, if there are pathological conditions of the reproductive tract, and the optimal time of mating. There have been many vaginal cytology studies and it’s clinical applications have been reported in many species. There have been few reports, however, about its use in buffalo. Buffaloes are considered poor breeders as their reproductive efficiency is adversely affected by certain limitations such as: late maturity, seasonality of breeding, silent heat coupled with poor expression of estrus, low conception rates and long intercalving period. Accurate heat detection is one of the key factors to increase the conception rate. Vaginal cytology in combination with other heat detection methods can help in this process. The objective of this review article is to focus on the use of the vaginal cytology technique for estrus detection and other applications in buffalo.