Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

Inefficacy of albendazole sulphoxide and ivermectin for the treatment of bovine parasitic otitis caused by rhabditiform nematodes

O objetivo deste estudo foi avaliar a eficácia do sulfóxido de albendazol administrado oralmente e da ivermectina "pour-on" no tratamento da otite parasitária bovina causada por nematóides rhabditiformes. Dezoito vacas Gir apresentando otite clínica foram divididas em três grupos de seis animais cada. O primeiro não recebeu tratamento (grupo controle). O segundo foi tratado com ivermectina "pour-on" a 0,05% na dose de 500µg/kg de peso vivo. O terceiro grupo foi tratado com sulfóxido de albendazol oral a 6% na dose 6,0mg/kg. Os condutos auditivos de todos os animais foram reexaminados nos dias 7 e 21 pós-tratamento. Os animais do grupo controle permaneceram infectados nos dias de observação. O tratamento com ivermectina não demonstrou eficácia alguma para os dias 7 e 21 pós-tratamento. O tratamento com sulfóxido de albendazol obteve 16,7 e 25% de eficácia nos dias 7 e 21, respectivamente. Mais estudos são necessários para determinação de tratamentos eficazes para tal doença parasitária, especialmente através de vias alternativas de administração, por causa de seu significante impacto na criação de Bos taurus indicus nas Regiões Tropical e Subtropical.