Kanarese buffaloes are medium built animals distributed in the South Kanara region on the west coast of South India. These buffaloes are dual purpose animals used mainly for agricultural operations in wet fields and for some milk production. They are famous for racing in water-bound fields. South Kanara buffaloes are moderate milk yielders producing about two to seven litres daily for a lactation period that ranged between 210 to 360 days. Genetic diversity analysis was performed using 10 microsatellite markers in a panel of 48 unrelated animals. The average number of alleles was estimated to be 6.30 with an average heterozygosity of 0.62 per locus. The population showed departure from the Hardy-Weinberg equilibrium at all of the 10 loci tested. The heterozygote deficiency was estimated as 9.2% suggesting the presence of considerable inbreeding in the population. The allele frequency distribution followed the normal L-shaped form suggesting that the breed had not encountered a genetic bottleneck in the recent past.