title: 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.
authors.affiliation: ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, Campinas, Sao Paulo, Brazil.
authors.forename: Dávila
authors.initials: D
authors.lastname: Zampieri
authors.forename: Vanessa G
authors.initials: VG
authors.lastname: Santos
authors.forename: Patrícia A C
authors.initials: PA
authors.lastname: Braga
authors.forename: Christina R
authors.initials: CR
authors.lastname: Ferreira
authors.forename: Daniela
authors.initials: D
authors.lastname: Ballottin
authors.forename: Ljubica
authors.initials: L
authors.lastname: Tasic
authors.forename: Andréa C
authors.initials: AC
authors.lastname: Basso
authors.forename: Bruno V
authors.initials: BV
authors.lastname: Sanches
authors.forename: José H F
authors.initials: JH
authors.lastname: Pontes
authors.forename: Bárbara Pereira
authors.initials: BP
authors.lastname: da Silva
authors.forename: Fabiana Fantinatti
authors.initials: FF
authors.lastname: Garboggini
authors.forename: Marcos N
authors.initials: MN
authors.lastname: Eberlin
authors.forename: Alessandra
authors.initials: A
authors.lastname: Tata
mesh_terms: D000818:Animals; D001419:Bacteria / Q000302:isolation & purification; D002417:Cattle / Q000196:embryology / Q000382:microbiology / Q000502:physiology; D015925:Cryopreservation / Q000592:standards / Q000662:veterinary*; D003470:Culture Media / Q000009:adverse effects / Q000032:analysis / Q000592:standards; D004622:Embryo, Mammalian / Q000382:microbiology*; D005260:Female; D005658:Fungi / Q000302:isolation & purification; D059471:In Vitro Oocyte Maturation Techniques / Q000592:standards / Q000662:veterinary; D008297:Male; D012661:Semen / Q000382:microbiology*; D055101:Semen Analysis / Q000592:standards; D012662:Semen Preservation / Q000009:adverse effects / Q000592:standards / Q000662:veterinary*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D003470:Culture Media
keywords: Bacteria; Culture media; In vitro production; Matrix-assisted laser desorption ionization mass spectrometry; Microorganisms; Semen