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      Detecção de Mycoplasma spp. e Ureaplasma diversum em vacas com distúrbios reprodutivos Translated title: Detection of Mycoplasma spp. and Ureaplasma diversum in cown with reproductive disorders

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          Abstract

          Foram utilizadas 112 amostras de muco vulvovaginal, coletados de vacas com distúrbios reprodutivos, para pesquisa de Mycoplasma e Ureaplasma. Para isolamentos, foram usados meios específicos para micoplasmas (SP-4) e para ureaplasmas. PCR genérica, PCR específica para Mycoplasma bovis e nested-PCR em tubo único para Ureaplasma diversum foram realizados com os DNAs extraídos das amostras. Mycoplasma spp. e U. diversum foram detectados em 12,5 e 25,0%, respectivamente. A PCR genérica resultou em reações positivas em 63,4% das amostras transportadas em SP-4 e em 69,6% das transportadas em meio de ureaplasma. M. bovis foi detectado, na PCR específica, em 9,8% das amostras e U. diversum, na nested-PCR, em 37,5%. Houve maior sensibilidade na metodologia da PCR quando comparada à técnica de cultivo para Mycoplasma e Ureaplasma.

          Translated abstract

          In the study, 112 samples of vulvovaginal mucus of cows bearing reproductive disturbance were investigated for Mycoplasma and Ureaplasma. Specific media for the culture of mycoplasmas (SP-4) and ureaplasmas were used. PCR with general primers, PCR specific for Mycoplasma bovis, and nested-PCR in a single tube for Ureaplasma diversum were performed to detect DNA of the sample. Mycoplasma spp. and U. diversum were isolated in 12.5 and 25.0%, respectively. With generic PCR, positive reaction was obtained in 63.4% of the samples transported in SP-4 and 69.6% in ureaplasma medium. M. bovis was detected in 9.8% of samples and nested-PCR in a single tube for U. diversum resulted in 35.0% of positive reaction. Results demonstrated increased sensitivity of PCR methodology compared with culture technique applied to the search of microorganisms of Mycoplasma and Ureasplasma genera.

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          Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification.

          Systematic computer alignment of mycoplasmal 16S rRNA sequences allowed the identification of variable regions with both genus- and species-specific sequences. Species-specific sequences of Mycoplasma collis were elucidated by asymmetric amplification and dideoxynucleotide sequencing of variable regions, using primers complementary to conserved regions of 16S rRNA. Primers selected for Mycoplasma pneumoniae, M. hominis, M. fermentans, Ureaplasma urealyticum, M. pulmonis, M. arthritidis, M. neurolyticum, M. muris, and M. collis proved to be species specific in the polymerase chain reaction. The genus-specific primers reacted with all mycoplasmal species investigated as well as with members of the genera Ureaplasma, Spiroplasma, and Acholeplasma. No cross-reaction was observed with members of the closely related genera Streptococcus, Lactobacillus, Bacillus, and Clostridium or with any other microorganism tested. On the basis of the high copy number of rRNA, a highly sensitive polymerase chain reaction assay was developed in which the nucleic acid content equivalent to a single organism could be detected.
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            Current status of the mollicute flora of humans.

            Of the more than 20 species of mollicutes reported to be present in human tissues, 15 have been isolated more than once and are currently thought to typify mollicutes of human origin. In the past decade a number of new and potentially significant mollicutes have been added to the list of species inhabiting humans. As our understanding of the human mollicute flora increases, diagnostic and clinical advances should permit the identification and control of those species that are significant pathogens in human disease.
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              Mycoplasma, Ureaplasma, and Acholeplasma infections of bovine genitalia.

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Journal
                abmvz
                Arquivo Brasileiro de Medicina Veterinária e Zootecnia
                Arq. Bras. Med. Vet. Zootec.
                Universidade Federal de Minas Gerais, Escola de Veterinária (Belo Horizonte )
                1678-4162
                December 2007
                : 59
                : 6
                : 1368-1375
                Affiliations
                [1 ] Universidade de São Paulo Brazil
                Article
                S0102-09352007000600003
                10.1590/S0102-09352007000600003
                63dc1bb7-1059-4aef-b2ce-3630a3f1111c

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0102-0935&lng=en
                Categories
                VETERINARY SCIENCES

                General veterinary medicine
                Mycoplasma bovis,Ureaplasma diversum,cow,culture,cultura,PCR,vaca
                General veterinary medicine
                Mycoplasma bovis, Ureaplasma diversum, cow, culture, cultura, PCR, vaca

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