Semen extenders or diluents are added to semen before cryopreservation for maintaining its fertilizing ability even after post thaw. Various types of semen extenders are commercially available for porcine, boar, bull, equine and humans. Bacteriospermia in human and animal samples appears to be concentration-time dependent which might affect quality, quantity and longevity of post ejaculated semen in either neat or with extended states. Antibiotics play a significant role in semen extender which ensures long shelf-life to spermatozoa. Also, it protects the female reproductive tract by hindering the transmission of invading pathogens. Although the use of antibiotics in semen extender is monitored by many authorities of respective concerned government, a lot of guidelines and experiments ensuring a proper usage in cryopreservation is in need. The use of antibiotics in semen extenders also pose a threat of antibiotic-resistant bacterial strains in artificial insemination centers (AI) as well as in assisted reproductive technology (ART) laboratories. Development of multi-drug resistant bacteria has been found to be the primary concern when mixture of a wide range of conventional antibiotics is used in semen extenders. Several potent strategies and critical point analysis are of urgent need to hinder the