Detecção do Grupo Mycoplasma mycoides por imunoperoxidase indireta (IPI) e PCR-REA em conduto auditivo de bovinos

Abstract

465Pesq. Vet. Bras. 30(5): 465-469, maio 2010 RESUMO.-O Grupo Mycoplasma mycoides (GMM) foi diagnosticado por PCR-REA e imunoperoxidase indireta (IPI) em amostras de lavados de conduto auditivo de bovinos no Estado do Rio de Janeiro, Brasil. 60 bovinos foram selecionados aleatoriamente. As lavagens foram feitas com uso de seringas estéreis contendo um volume de 60 mL de solução salina tamponada (PBS pH 7.2). As amostras obtidas foram estocadas em glicerol (1:2) e congeladas a -20 o C até uso. Estas amostras foram diluídas até 10 -5 e repicadas em meio Hayflick modificado, sólido e líquido, Mycoplasma mycoides cluster (MMC) was diagnosed by polimerase chain reactionrestriction endonuclease analysis (PCR-REA) and indirect immunoperoxidase (IPI), both, carried out in flushing from external ear canal, collected from bovine at slaughter time in the State of Rio de Janeiro, southeastern, Brazil. A total of 60 bovines were randomly selected. Sterile syringes (60mL) loaded with buffer solution (PBS, pH 7.2) were used for the ear canal flushing. The obtained samples were stored in glycerol (1:2) and frozen at -20 o C until use. These specimens were diluted up to 10 -5 , inoculated in liquid and solid modified Hayflick´s media and incubated at 37 o C for 2-3 days. The plates were kept in a microaerophilia condition and examined every two days under a stereomicroscope for the presence of typical colonies "fried-egg". In this study, 35 strains selected in agreement with their biochemistry and physiologic proprieties, were used. From the 60 cultivated samples, 48 (80.00%) were positive for Mycoplasma spp. Under IPI the prevalence obtained for MMC was 20.0% (12/60) while by PCR-REA it was 41.7% (25/60). The IPI typing of these isolates resulted in 58.3% (7/12) for M.mycoides mycoides LC and 41.7% (5/12) for M. capricolum. PCR-REA for MMC was confirmed by the amplicon size of 785bp, compatible with this group. The Kappa value for the association between these two tests was 0.14 (p>0.05). After restriction analysis with AluI in all MMC strains the fragments size obtained were of 81, 98, 186 and 236bp, but not of 370bp that is compatible with Mycoides mycoides mycoides SC of bovine type. The presence of mycoplasmas species in the ear canal of asymptomatic bovines represent a risk of subsequent propagation of Mycoplasma spp. among bovine herds in Brazil. Detecção do GrupoINDEX TERMS: Mycoplasma mycoides , flushing ear, cattle, PCR-REA diagnostic.

Anti-mycoplasma activity of Curcuma longa extracts and your isolated compound, the curcumin

“…The Mycoplasmas are part of the Mollicutes class, a large group of peculiar microorganisms responsible for a series of diseases in animals, plants and in humans, being notable for causing sexually transmitted diseases (STDs) and respiratory infections (1) . These microorganisms are capable of self-replication (2) , whose major phenotypic characteristics are the absence of cell wall and its reduced genome (3) . Additionally, a significantly increased incidence of infections, more likely in immunocompromised patients, causing opportunistic infections associated with acquired immunodeficiency syndrome (AIDS), cancer chemotherapy and transplantation.…”