Microorganisms in cryopreserved semen and culture media used in the in vitro production (IVP) of bovine embryos identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS)

Abstract

Commercial cattle breeders produce their own herd offspring for the dairy and beef market using artificial insemination. The procedure involves sanitary risks associated with the collection and commercialization of the germplasm, and the in vitro production and transfer of the bovine embryos must be monitored by strict health surveillance. To avoid the spreading of infectious diseases, one must rely on using controlled and monitored germplasm, media, and reagents that are guaranteed free of pathogens. In this article, we investigated the use of a new mass spectrometric approach for fast and accurate identification of bacteria and fungi in bovine semen and in culture media employed in the embryo in vitro production process. The microorganisms isolated from samples obtained in a commercial bovine embryo IVP setting were identified in a few minutes by their conserved peptide/protein profile, obtained applying matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), matched against a commercial database. The successful microorganisms MS identification has been confirmed by DNA amplification and sequencing. Therefore, the MS technique seems to offer a powerful tool for rapid and accurate microorganism identification in semen and culture media samples.

Bacteriospermia and Sperm Quality of Cryopreserved Bull Semen Used in Artificial Insemination of Cows in South Wollo Zone, Ethiopia

“…In other species, Staphylococcus epidermidis in boar semen [43] and CoNS in human semen samples [45] were identified. e discrepancy found in the present study when compared to the above authors may be due to differences in sample size, bacteriological techniques [24], and age and breeds of animals [17].…”

“…us, one of the main factors contributing to AI failure is the contamination of the germplasm with pathogens, which eventually cause loss of fertility [20]. Most common microorganisms that contaminated bull semen were from species of Corynebacterium, Staphylococcus, Micrococcus, Bacillus, Escherichia, Proteus, Pseudomonas, Klebsiella, Streptococcus, Citrobacter, Enterobacter, and Stenotrophomonas [21][22][23][24][25]. ese pathogens, in one way or other, may infect inseminated females or contribute to a rapid deterioration in sperm quality [26].…”

Current and alternative trends in antibacterial agents used in mammalian semen technology

“…Although one of the benefits of AI is to reduce the spread of diseases, this technique may allow the pathogens to be easily disseminated through the transmission of contaminated semen (Foote, 2002). The semen can be contaminated particularly during the process of its collection or cryopreservation if good practices are not adopted (Zampieri et al, 2013). Moreover, the presence of pathogens can damage the sperms (Diemer et al, 2000), adversely affecting the quality of the semen subjected to cryopreservation.…”

Colloids: Applications in Sperm Preparation for Assisted Reproduction

“…However, antibiotic use in general can lead to the development of microbial resistance, which can be passed on to other animals and also between humans and animals. Recent reports indicate that some microorganisms may be resistant to the antibiotics used in semen extenders for livestock [52][53][54]. Therefore, it might be prudent to develop alternatives to antimicrobials in semen extenders [55], although a change in the legislation would be required in the event that antibiotics are not added.…”