Utilization of the cauda epididymis as an alternative spermatozoa source for assisted reproductive technology (ART) has been adopted in many species. This technique is useful to preserve genetic diversity in endangered species, and from animals experiencing sudden death or pathology resulting in an inability to ejaculate. This technique has been used for several decades, but data about the distribution of the sex ratio of the offspring produced is limited. In a preliminary artificial insemination (AI) study concerning using frozen-thawed epididymal spermatozoa from the spotted buffalo, a skewed sex ratio was observed. Results of this study showed that mean progressive motility, viability, and membrane integrity of fresh epididymal spermatozoa were 73.3%, 85.4%, and 86.2%, decreasing to 43.3%, 66.6%, and 66.9% after thawing, respectively. We got high and with a significantly higher number of female (12; 85.7%) compare to male (2; 14.3%) offspring (P<0.05). These interesting preliminary results indicate that a further study is needed.