Molecular characterization of Mycobacterium bovis infection in cattle and buffalo in Amazon Region, Brazil

Abstract

The aim of this study was to characterize Mycobacterium bovis from cattle and buffalo tissue samples, from two Brazilian states, and to analyse their genetic diversity by spoligotyping. Tissue samples from tuberculosis suspect animals, 57 in Amazonas State (12 cattle and 45 buffaloes) and six from Pará State (5 cattle and one buffalo) from slaughterhouses under State Veterinary Inspection, were isolated in culture medium Stonebrink. The positive cultures were confirmed by PCR and analysed by the spoligotyping technique and the patterns (spoligotypes) were identified and compared at the Mycobacterium bovis Spoligotype Database (http://www.mbovis.org/). There was bacterial growth in 44 (69.8%) of the tissues of the 63 animals, of which PCR for region of differentiation 4 identified 35/44 (79.5%) as Mycobacterium bovis. Six different spoligotypes were identified among the 35 Mycobacterium bovis isolates, of which SB0295, SB1869, SB0121 and SB1800 had already been described in Brazil, and SB0822 and SB1608 had not been described. The most frequent spoligotype in this study (SB0822) had already been described in buffaloes in Colombia, a neighbouring country of Amazonas state. The other identified spoligotypes were also described in other South American countries, such as Argentina and Venezuela, and described in the Brazilian states of Rio Grande do Sul, Santa Catarina, São Paulo, Minas Gerais, Mato Grosso do Sul, Mato Grosso and Goiás, indicating an active movement of Mycobacterium bovis strains within Brazil.

Epidemiological Study of Mycobacterium bovis Infection in Buffalo and Cattle in Amazonas, Brazil

“…The apparent prevalence in herds composed only by cattle, by buffalo and cattle, and only by buffalo was respectively, 45.4, 66.7, and 80% ( Table 5). As reported before there were no significant differences between LST samples and no LST (42).…”

Genetic Diversity and Potential Paths of Transmission of Mycobacterium bovis in the Amazon: The Discovery of M. bovis Lineage Lb1 Circulating in South America

“…A total of 24 M. bovis isolates were selected, representing one herd of each municipality involved in two previous studies ( 6 , 20 ) that obtained a total of 63 M. bovis isolates from Amazonas State (12 from cattle and 45 from buffalo) and from Pará State (5 from cattle and 1 from buffalo). The selection of M. bovis isolates maintained the proportionality according to species (cattle and buffalo) and the local prevalence from the previous studies ( 6 , 20 ).…”

“…A total of 24 M. bovis isolates were selected, representing one herd of each municipality involved in two previous studies ( 6 , 20 ) that obtained a total of 63 M. bovis isolates from Amazonas State (12 from cattle and 45 from buffalo) and from Pará State (5 from cattle and 1 from buffalo). The selection of M. bovis isolates maintained the proportionality according to species (cattle and buffalo) and the local prevalence from the previous studies ( 6 , 20 ). Accordingly, these isolates were from tissue samples of 8 cattle and 16 buffalos collected at the slaughterhouse from herds with or without known tuberculin skin test (TST) status originating from 12 different municipalities: Alenquer ( n = 1), Apui ( n = 1), Autazes ( n = 1 ), Careiro da Varzea ( n = 1), Itacoatiara ( n = 2), Manacapuru ( n = 1), Novo Ceu ( n = 8), Parintins ( n = 3), Prainha ( n = 2), Presidente Figueiredo ( n = 1), and Urucara ( n = 3).…”

“… Geographic origin and host species of the Mycobacterium bovis isolates from municipalities of Lower Amazon River Basin, Brazil. Isolates were selected from 63 M. bovis strains previously isolated from cattle and buffalo ( 6 , 20 ). …”

“…In addition, the knowledge regarding strain diversity within host species has special contribution in areas under risk of zoonotic TB occurrence, thereby providing new insights in strain distribution that may help establishing strategic measures for TB control and prevention ( 18 , 19 ). In Brazil, few and dispersed molecular epidemiologic studies of M. bovis have been reported ( 20 – 24 ), and the WGS characterization of the pathogen is just starting to be unraveled ( 24 – 26 ). While few studies focused on areas of high dairy herd productivity ( 25 , 26 ), a recent study evaluating M. bovis genomes obtained from buffalo and cattle of the Marajó Island, Northern Brazil, was performed and showed the existence of a monophyletic group without CC classification ( 24 ).…”