Genotyping of Ureaplasma diversum isolates using pulsed-field electrophoresis

Molecular characterization of ureaplasmas isolated from reproductive tract of goats and sheep from Brazil

“…Marois 11 and Buzinhani 12 reported that 100% of the isolates of Mycoplasma synoviae and U. diversum , respectively, did not allow for typing with PFGE methodology. Buzinhani 12 mentioned the absence of fragments due to the low DNA content. In fact, ureaplasma grows fast, but is sensitive to pH shift and is fragile to osmotic shock due to the absence of the cell wall.…”

“…Just after pH shifting of each broth, they were centrifuged for 75 min at 16,300 ×  g , then washed twice with PBS, followed by centrifugation for 25 min at 20,600 ×  g before being resuspended in 400 μL of Tris-EDTA (10 mM Tris–HCl, 1 mM EDTA, pH 8.0). The DNA from the pelleted cells was extracted in agar blocks and digested with Sal I, as described by Marois 11 and Buzinhani 12 . The PFGE was performed using 1.2% ultrapure agarose gel (Bio-Rad) in 0.5% TBE buffer (90 mM Tris–borate, and 1 mM EDTA) at 14 °C in a CHEF Mapper DR-III (Bio-Rad).…”

Invasion of Ureaplasma diversum in bovine spermatozoids

“…The semen was previously tested for mollicute contamination by PCR [32]. The frozen-thawed semen was layered in microtubes with a Percoll gradient (1:1 [vol/vol] mixture of 90%/45%) and centrifuged at 9,000 xg for 5 min [33].…”

Estudo da variabilidade genética e dos fatores de virulência de isolados de Ureaplasma diversum.

“…As cepas e isolados foram divididas em sete grupos, definidos de acordo com perfil de polimorfismos similares ou idênticos. Pettersson et al (1998) As cepas e isolados das propriedades 1, 2, 3, 4 e 5 do presente estudo, foram previamente submetidas a análise de variação genotípica por meio das metodologias de PFGE e AFLP (BUZINHANI et al, 2007a;BUZINHANI et al, 2007b , M.…”

“…Os ureaplasmas bovinos são considerados molecularmente e sorologicamente heterogêneos. Buzinhani et al (2007aBuzinhani et al ( , 2007b Experimentalmente, este microrganismo não causou alterações ultraestruturais na mórula bovina (BRITTON et al, 1987), mas confirmou-se a presença de ureaplasmas na superfície externa da zona pelúcida, podendo ser transmitido para recipientes durante a transferência de embrião (BRITTON et al, 1989). Smits et al (1994) sugeriram que U. diversum altera a integridade de células do oviduto, contribuindo para a deficiência de implantação do embrião bovino e seu desenvolvimento.…”