Antimicrobials in a rabbit semen extender: effects on reproduction
Keywords:Rabbit, Semen, Dilution media, Bacterial charges, Motility, Fertility
The use of fluoroquinolone antibiotics was legally restricted by the European Commission in March 2019. Since the extender for rabbit semen Galap® contained this antibiotic, it became necessary to modify it. The purpose of this study was to search for another molecule, based on its antimicrobial activity and also on the conservation, motility and fertility of semen diluted with the new extender. Several bacterial strains were isolated from 10 poor-quality ejaculates, including Enterococcus spp., Staphylococcus aureus and Proteus spp. They were then tested for antimicrobial susceptibility. Out of 15 antibiotics evaluated, gentamicin was the one targeting such bacteria. In vivo tests were then carried out to assess the effects of this antibiotic change on sperm parameters. Up to 26 pools of good quality semen (total motility >70%) were diluted in original Galap® or in this extender with gentamicin. Ejaculates were analysed on the day (D) of collection and up to 6 d of storage at +17°C. After 24 h storage, the motility in the new extender was reduced by 7.7% compared to the original; this decrease did not worsen by storing. After 6 d of storage, no difference between the two media was detected. A total of 360 females were then inseminated with pools of 4 to 5 good quality semen diluted 1:10 in both extenders, following the routine protocol of rabbit semen processing centres. There was no difference in female fertility or prolificacy between both formulations. In conclusion, these preliminary results suggest that the addition of gentamicin to this extender is useful and also has no adverse effect on fertility or prolificacy.
Althouse G.C. 2008. Sanitary procedures for the production of extended semen. Reprod. Domest. Anim. 43 Suppl 2: 374-378. https://doi.org/10.1111/j.1439-0531.2008.01187.x
Althouse G.C., Kuster C.E., Clark S.G., Weisiger R.M. 2000. Field investigations of bacterial contaminants and their effects on extended porcine semen. Theriogenology, 53: 1167-1176. https://doi.org/10.1016/S0093-691X(00)00261-2
Althouse G.C., Lu K.G. 2005. Bacteriospermia in extended porcine semen. Theriogenology, 63: 573-584. https://doi.org/10.1016/j.theriogenology.2004.09.031
Althouse G.C., Pierdon M.S., Lu K.G. 2008. Thermotemporal dynamics of contaminant bacteria and antimicrobials in extended porcine semen. Theriogenology, 70: 1317-1323. https://doi.org/10.1016/j.theriogenology.2008.07.010
Aurich C., Spergser J. 2007. Influence of bacteria and gentamycin on cooled-stored stallion spermatozoa. Theriogenology, 67: 912-918. https://doi.org/10.1016/j.theriogenology.2006.11.004
Berendsen B., Elbers I., Stolker L. 2011. The stability of antibiotics in matrix and reference solutions determined using a straight-forward procedure applying mass spectrometric detection. Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess., 28: 1657-1666. https://doi.org/10.1080/19440049.2011.604045
Bresciani C., Cabassi C.S., Morini G., Taddei S., Bettini R., Bigliardi E., Ianni F.D., Sabbioni A., Parmigiani E. 2014. Boar Semen Bacterial Contamination in Italy and Antibiotic Efficacy in a Modified Extender. Ital. J. Anim. Sci., 13: 3082. https://doi.org/10.4081/ijas.2014.3082
Brun J.M., Theau-Clément M., Bolet G. 2002. The relationship between rabbit semen characteristics and reproductive performance after artificial insemination. Anim. Reprod. Sci., 70: 139-149. https://doi.org/10.1016/S0378-4320(01)00197-X
Bussalleu E., Yeste M., Sepúlveda L., Torner E., Pinart E., Bonet S. 2011. Effects of different concentrations of enterotoxigenic and verotoxigenic E. coli on boar sperm quality. Anim. Reprod. Sci., 127: 176-182. https://doi.org/10.1016/j.anireprosci.2011.07.018
Daniel N., Renard J.P. 2010. Artificial Insemination in Rabbits. Cold Spring Harb Protoc., https://doi.org/10.1101/pdb.prot5358
Duracka M., Lukac N., Kacaniova M., Kantor A., Hleba L., Ondruska L., Tvrda E. 2019. Antibiotics versus Natural Biomolecules: The Case of in vitro Induced Bacteriospermia by Enterococcus faecalis in Rabbit Semen. Molecules 24: 4329. https://doi.org/10.3390/molecules24234329
Francisco E.M. 2018. Fluoroquinolone and quinolone antibiotics: PRAC recommends new restrictions on use following review of disabling potentially long-lasting side effects. Available at: https://www.ema.europa.eu/en/news/fluoroquinolonequinolone-antibiotics-prac-recommends-new-restrictionsuse-following-review Accessed December 2020.
Fraczek M., Kurpisz M. 2015. Mechanisms of the harmful effects of bacterial semen infection on ejaculated human spermatozoa: potential inflammatory markers in semen. Folia Histochem. Cytobiol. 53: 201-217. https://doi.org/10.5603/fhc.a2015.0019
Gadea J. 2003. Semen extenders used in the artificial insemination of swine: Review. Span. J. Agric. Res. 17-28. https://doi.org/10.5424/sjar/2003012-17
Gloria A., Contri A., Wegher L., Vignola G., Dellamaria D., Carluccio A. 2014. The effects of antibiotic additions to extenders on fresh and frozen-thawed bull semen. Anim. Reprod. Sci., 150: 15-23. https://doi.org/10.1016/j.anireprosci.2014.08.012
Holtz W., Foote, R.H. 1978. Composition of rabbit semen and the origin of several constituents. Biol. Reprod., 18: 286-292. https://doi.org/10.1095/biolreprod18.2.286
Hudzicki J. 2009. Kirby-Bauer Disk Diffusion Susceptibility Test Protocol. Washington, DC: American Society for Microbiology, 23 pp.
International Rabbit Reproduction Group (IRRG). 2005. Recommendations and guidelines for applied reproduction trials with rabbit does. World Rabbit Sci., 13: 147-164. https://doi.org/10.4995/wrs.2005.521
Jarvis S., Day J.E.L., Reed B. 2005. BSAS Ethical Policy British Society of Animal Science Ethical guidelines for research in animal science. In Proc. of the British Society of Animal Science, 247-253. https://doi.org/10.1017/S1752756200011571
Koppers A.J., De Iuliis G.N., Finnie J.M., McLaughlin E.A., Aitken R.J. 2008. Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa. J. Clin. Endocrino. Metabo., 93: 3199-3207. https://doi.org/10.1210/jc.2007-2616
Lavara R., Mocé E., Lavara F., Viudes de Castro M.P., Vicente J.S. 2005. Do parameters of seminal quality correlate with the results of on-farm inseminations in rabbits? Theriogenology, 64: 1130-1141. https://doi.org/10.1016/j.theriogenology.2005.01.009
Lebas F., Coudert P., De Rochambeau H. 1997. The Rabbit: husbandry, health, and production. Food and Agriculture Organization of the United Nations. Rome, Italy.
Maes D., Nauwynck H., Rijsselaere T., Mateusen B., Vyt P., de Kruif A., Van Soom A. 2008. Diseases in swine transmitted by artificial insemination: an overview. Theriogenology, 70: 1337-1345. https://doi.org/10.1016/j.theriogenology.2008.06.018
Maroto Martín L.O., Muñoz E., De Cupere F., Van Driessche E., Echemendia-Blanco D., Rodríguez J., Beeckmans S. 2010. Bacterial contamination of boar semen affects the litter size. Anim. Reprod. Sci., 120: 95-104. https://doi.org/10.1016/j.anireprosci.2010.03.008
Mercier P., Rideaud P. 1990. Bactériologie du sperme frais de lapin - Etude préliminaire. INRAE Productions Animales 3: 215-221. https://doi.org/10.20870/productions-animales.19220.127.116.1178
Morrell J., Wallgren M. 2014. Alternatives to Antibiotics in Semen Extenders: A Review. Pathogens, 3: 934-946. https://doi.org/10.3390/pathogens3040934
R Development Core Team. 2014. R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria.
Theau-Clément M., Ailloud E., Sanchez A., Saleil G., Brun J.M. 2016. Relationships between rabbit semen characteristics and fertilising ability after insemination. Animal, 10: 426-431. https://doi.org/10.1017/S1751731115002372
Waberski D., Luther A.M., Grünther B., Jäkel H., Henning H., Vogel C., Peralta W., Weitze K.F. 2019. Sperm function in vitro and fertility after antibiotic-free, hypothermic storage of liquid preserved boar semen. Sci. Rep., 9: 1-10. https://doi.org/10.1038/s41598-019-51319-1
Copyright (c) 2022 Charlène Rouillon, Sabine Camugli, Olivier Carion, Arantxa Echegaray, Guy Delhomme, Eric Schmitt
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
This journal is licensed under a "Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)".