ARTIFICIAL INSEMINATION IN FARM ANIMALS - Phenix-Vet

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12Artificial Insemination in Farm Animalsto them. It is intended to use the antibodies to aggregate spermatozoa bearing a specific sexchromosome, thus enabling them to be removed from the general population.A combination of ARTs would also be relevant for sperm sexing. Thus, the speed of flowsorting can be increased by first removing the dead and dying spermatozoa from thepopulation, for example by density gradient centrifugation or single layer centrifugation.Such a combination may increase the “sortability” of sperm samples. Sufficient sexedspermatozoa may be obtained from flow sorting for IVF, thus generating embryos orblastocysts for subsequent transfer. However, methods of speeding up the selection processare needed if flow cytometry is to become useful for species other than the bovine.6.4 Sperm cryopreservationAs previously mentioned, the ability of cryopreserved spermatozoa to retain their fertilizingability varies widely between species. New cryoextenders and new protocols are beingdeveloped constantly in an effort to address this issue. One recent advance has been theintroduction of dimethylsulphoxide and the amides formamide and dimethylformamide ascryoprotectants, in place of glycerol. These molecules seem to function better than glycerolfor some individuals whose spermatozoa do not freeze well, for example, some stallions.One explanation for this observation is that these molecules are smaller than glycerol andtherefore may cause less damage when they penetrate the sperm membrane. However, nomethod appears to be universally successful within one species. As far as turkeyspermatozoa are concerned, it seems that the development of a successful freezing methodwill require more than new cryoprotectants and additives (Holt, 2000).6.5 Removal of viruses from ejaculatesViral infectivity can be removed from the semen of patients with viral infections such asHIV and hepatitis, by a sequential method of sperm preparation i.e. centrifugation on adensity gradient followed by a “swim-up” (reviewed by Englert et al., 2004). Spermatozoafrom virally infected men prepared by this method have been used in assisted reproductionattempts, apparently without sero-conversion of mothers or children. However, somestudies with HIV report that density gradient centrifugation alone will not remove all viralinfectivity (Politch et al., 2004). Since spermatozoa may function as vectors for viruses (Chanet al., 2004), further work is required to investigate how closely different viral particles areassociated with the sperm membrane with putative carry-over during processing. Thedouble method of processing has also been successful in removing equine arteritis virusfrom an infected stallion ejaculate in a preliminary study (Morrell & Geraghty, 2006). SLCtogether with swim-up was used to reduce viral infectivity from boar semen spiked withporcine circo virus 2 (Blomqvist et al., 2011).6.5 AI in conservation biologyIt has been suggested that AI and other forms of ART could be useful for geneticconservation and preservation of rare breeds. Many of these technologies have beensuccessful to some degree in a research setting, but none have produced results sufficient toeffect population-wide improvements in genetic management (Morrow et al., 2009).Cryopreservation of semen has been the most widely applied ART in this respect, but muchof the frozen semen in so-called gene banks has never been tested for fertility. A lack ofsuitable females or dearth of knowledge about the reproductive biology of the species
Artificial Insemination: Current and Future Trends 13involved may contribute to this deficit. However, long-term storage of frozen gametes ofunknown fertility is not a sustainable policy for the conservation of rare breeds andendangered species. The development of in vitro methods of testing sperm fertility wouldcontribute considerably to conservation efforts. Since the semen quality in these animalsmay be poor (Gamboa et al., 2009), techniques such as SLC of samples prior to AI could beof considerable benefit in conservation breeding.7. ConclusionAI revolutionized animal breeding in the 20th century, particularly in combination withsperm cryopreservation. The AI industry has developed dramatically in most domesticspecies in the last few decades and its use is now widespread in intensive animalproduction. The development of other associated technologies, such as sperm selection andsex selection, are predicted to create powerful tools for the future, both for domesticlivestock breeding and for the purposes of conservation. AI will continue to play a role infertility treatment for human patients, although it may be superseded by IVF or ICSI. It hasbeen suggested that AI (in animals) is entering a new era where it will be used for theefficient application of current and new sperm technologies (Roca, 2006). Excitingpossibilities are offered by emerging techniques, such as Single Layer Centrifugation, forimproving sperm quality in AI doses as well as for increasing sperm survival duringcryopreservation.8. ReferencesBellinge, B.S., Copeland, C.M., Thomas, T.D., Mazzucchelli, R.E., O’Neil, G., Cohen M.J.(1986). The influence of patient insemination on the implantation rate in an in vitrofertilization and embryo transfer program. Fertil Steril,, Vol. 46, 2523–2526.Chan, P.J., Su, BC., Kalugdan, T., Seraj, J.M., Tredway, D.R., King, A. (1994). Humanpapillomavirus gene sequences in washed human sperm deoxyribonucelic acid.Fertil Steril., Vol. 61, pp. 982-985.Englert, Y., Lesage, B., van Vooren, J.P, Liesnard, C., Place, I., Vannin, A.S., Emiliani, S.,Delbaere, A. (2004). Medically assisted reproduction in the presence of chronic viraldisease. Hum. Reprod. Update, Vol. 10, pp. 149-162.Gamboa, S., Machado-Faria M,, Ramalho-Santos J. (2009). Seminal traits, suitability forsemen preservation and fertility in the native Portuguese horse breeds Puro SangueLusitano and Sorraia: Implications for stallion classification and assistedreproduction. Anim Reprod Sci.Vol.113, pp.102-113.Gutsche, S., Wolff, M., von Strowitzki, T., Thaler CJ. (2003). Seminal plasma induces mRNAexpression of IL-1β, IL-6 and LIF in endometrial epithelial cells in vitro. Mol HumReprod, Vol. 9, pp. 785–791.Holt, WV. (2000). Basic aspects of frozen storage of semen. Anim. Reprod. Sci., Vol. 62, 3–22.Leboeuf, B., Delgadillo, JA., Manfredi, E., Piacere, A., Clement, V., Martin, P., Pellicer, M.,Boue, P., de Cremoux, R. (2008) Management of Goat Reproduction andInsemination for Genetic Improvement in France Reprod Dom Anim, Vol. 43 (Suppl.2), pp. 379–385
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ARTIFICIAL INSEMINATION IN FARM ANIMALS - Phenix-Vet

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