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Biopreserv Biobank. 2019 Dec 2. doi: 10.1089/bio.2019.0054. [Epub ahead of print]

Honey as an Alternative to Antibiotics for Cryopreservation of Nili-Ravi Buffalo Bull Spermatozoa.

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Department of Zoology, Pir Mehr Ali Shah - Arid Agriculture University, Rawalpindi, Pakistan.
Department of Wildlife Management, Pir Mehr Ali Shah - Arid Agriculture University, Rawalpindi, Pakistan.
Division of Science and Technology, Township, Department of Zoology, University of Education, Lahore, Pakistan.
Academic Unit of Reproductive and Developmental Medicine, University of Sheffield, Sheffield, United Kingdom.


The antimicrobial properties of honey have stimulated interest in evaluating it as an alternative to antibiotics for cryopreserved buffalo semen. Acacia nilotica, Brassica campestris and Ziziphus jujuba honey were analyzed and Z. jujuba honey was found suitable in terms of quality and purity. Buffalo semen (24 ejaculates) was studied for in vitro dose tolerability to Z. jujuba honey (0.1%-1%), and up to 0.2% (v/v) was not toxic to buffalo spermatozoa. Afterward, semen from three bulls (24 ejaculates) was cryopreserved (four replicates) in tris-citric egg yolk extender supplemented with 0.1% or 0.2% honey, with or without streptomycin-penicillin (SP); extender with SP used as a control. After dilution and cooling, extender without antibiotics but with 0.2% honey was better (p < 0.05) than control in terms of sperm motility and plasma membrane integrity. After thawing, the extenders containing 0.1% honey with antibiotics and extender having 0.2% honey without antibiotics consistently yielded good results in terms of all parameters studied compared to control and other extenders. The extender containing 0.2% honey without antibiotics was better (p < 0.05) in terms of total aerobic bacterial count. In conclusion, 0.2% honey improves the post-thaw quality of buffalo spermatozoa and can replace the use of antibiotics in extender through its antimicrobial activity.


Nili-Ravi buffalo; Ziziphus jujuba; antimicrobial activity; honey; semen cryopreservation


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