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Quality semen production remains the main focus and objective of semen processing laboratories throughout the world. Bacterial and other microbial contaminants affect the semen quality and hence the fertility, and also lead to reproductive disorders as well as lower conception rates and increased embryonic mortality, abortion and other complications in females. Microbial contamination affects the semen adversely, by exerting direct spermicidal effect, formation of reactive oxygen species, toxin production, adherence with spermatozoa, deriving nutrients and oxygen from the medium and thus competing with spermatozoa for the factors of growth and normal functioning. Despite hygienic measures, several ubiquitous and opportunistic microbes find their ways into semen during collection, processing, and storage of semen, and survive even during freezing. Stringent sanitary precautions are therefore required at every step of collecting semen and its processing. Preputial cavity is considered as main source of semen contaminating microorganisms. Flushing the preputial cavity with normal saline or any suitable liquid combination with antimicrobial activity, prior to semen collection reduces the microbial load and thereby improves the semen quality.

Aim: To study the effect of preputial washing on bacterial load, preservability and semen quality in Murrah buffalo bulls Materials and Methods: A total of 36 collections of three Murrah buffalo bulls maintained at Artificial Breeding Research Centre, ICAR-National Dairy Research Institute, Karnal, were collected at weekly intervals from each bull without preputial washing and latter ejaculates from same bull with preputial washing by infusing normal saline (0.85%), KMnO4 (0.02%) and savlon (2.0%) to first, second and third bull, respectively. The microbial load and semen quality were evaluated during different hours of storage at refrigerated temperature (0, 24 and 48 h) and after thrawing of cryopreserved (at −196°C) semen. Results: The results of preservation of semen at refrigerated temperature showed that bacterial load was markedly lower in ejaculates of bulls subjected to preputial washing. Semen preserved at refrigerator temperature and cryopreserved, the effect of washing solution was significant for individual motility (IM), non-eosiniphilic count, hypo-osmotic swelling reactivity (HOST), total plate count (TPC) and acrosome integrity. KMnO4 was found to be the best in lowering bacterial load, sperm abnormalities and in improving semen quality such as motility, non-eosinophilic count, HOST and acrosome integrity even up to 48 h of preservation and cryopreserved semen. Effect of duration of preservation and stage of cryopreservation was also significant for IM, non-eosiniphilic count, HOST, sperm abnormalities and acrosome integrity. Conclusion: Overall the results suggested that preputial washing with KMnO4 solution improved the semen quality and reduced microbial load of Murrah buffalo bull’s semen preserved at refrigerated temperature and cryopreservation.