Molecular Typing of Mycobacterium bovis from Cattle Reared in Midwest Brazil

Abstract

Mycobacterium bovis is the causative agent of bovine tuberculosis (BTB), the pathogen responsible for serious economic impact on the livestock sector. In order to obtain data on isolated M. bovis strains and assist in the control and eradication program for BTB, a cross sectional descriptive molecular epidemiology study in the Brazilian Midwest was conducted. Through spoligotyping and 24-loci MIRU-VNTR methods, 37 clinical isolates of M. bovis circulating in the region were analyzed, 10 isolated from the state of Mato Grosso, 12 from the state of Mato Grosso do Sul and 15 from the state of Goiás. The spoligotyping analysis identified 10 distinct M. bovis profiles (SB0121 n = 14, SB0295 n = 6, SB0140 n = 6, SB0881 n = 3, SB1144 n = 2, SB1145 n = 2, SB0134 n = 1, SB1050 n = 1, SB1055 n = 1, SB1136 n = 1) grouped in six clusters and four orphan patterns. The MIRU-VNTR 24-loci grouped the same isolates in six clusters and 22 unique orphan patterns, showing higher discriminatory power than spoligotyping. When associating the results of both techniques, the isolates were grouped in five clusters and 24 unique M. bovis profiles. Among the 24-loci MIRU-VNTR evaluated, two, ETR-A and QUB 11b loci, showed high discriminatory ability (h = ≥ 0.50), while MIRU 16, MIRU 27, ETR-B, ETR-C, Mtub21 and QUB 26 loci showed moderate ability (h = 0.33 or h = 0.49) and were the most effective in evaluating the genotypic similarities among the clinical M. bovis isolate samples. Herein, the 29 patterns found amongst the 37 isolates of M. bovis circulating in the Brazilian Midwest can be due to the animal movement between regions, municipalities and farms, thus causing the spread of various M. bovis strains in herds from Midwest Brazil.

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

“…The spoligotype SB0121 is considered the most prevalent in several studies and the most frequent in Brazil (Zumárraga et al, ) and was observed in two (5.7%) buffaloes in the municipality of Autazes, as well as described in other studies, in the states of Bahia (Filho et al, ), Paraíba (Parreiras et al, ), Mato Grosso, Mato Grosso do Sul (Cazola et al, ), Goiás (Carvalho et al, ), Minas Gerais (Parreiras et al, ) and Rio Grande do Sul (Ramos et al, ). Outside of Brazil, SB0121 has been described in Colombia (Jojoa‐Jojoa et al, ), Argentina, Venezuela (Zumárraga et al, ), Mexico (Reyes et al, ), Portugal (Reis, ) and France (Hauer et al, ).…”

“…Other spoligotypes considered unusual were observed in this study, as SB1869, which had already been described in São Paulo by Rocha (Rocha, 2013), was identified in five buffaloes (14.3%) in (Table 1). Also considered unusual, SB1608 and SB1800 spoligotypes were isolated in this study, the spoligotype SB1608 isolated in buffalo in the municipality of Parintins had not previously been described in Brazil and was described in wild animals in Portugal (Carvalho et al, 2016), SB1800 was described in Brazil (www.mbovis.org) and identified in cattle in Apuí-AM (Table 1).…”

“…SB0295 was the second most frequent spoligotype observed, with nine buffaloes in Amazonas, and was most frequently found in animals without LST (7/9). This spoligotype is considered the second most frequent in Brazil (Zumárraga et al, ), described in the state of Paraíba (Higino et al, ), Bahia (Filho et al, ), Mato Grosso, Goiás (Carvalho et al, ), Mato Grosso do Sul, Santa Catarina (Parreiras et al, ) and the state of São Paulo (Rocha, ), but not described or identified in the states of Amazonas and Pará. Outside of Brazil, it was described in buffaloes in Argentina (Zumárraga et al, ) and alpacas ( Lama pacos ) (García‐Bocanegra et al, ), and wild boars ( Sus scrofa ) in Spain (García‐Bocanegra et al, ).…”

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“…bovis são menos abundantes que para M. tuberculosis, no entanto a tipagem VNTR-MIRU foi usada em muitos países para estudos epidemiológicos de M. bovis (RORING et al, 2002;SKUCE et al, 2002;RORING et al, 2004;HILTY et al, 2005;ALLIX et al, 2006;JEON et al, 2008;ROMERO et al, 2008;BONIOTTI et al, 2009;Mc LERNON et al, 2010;SAHRAOUI et al, 2010;MANDAL et al, 2011;FIGUEIREDO et al, 2012;PARREIRAS et al, 2012;ROCHA et al, 2012;SUN et al, 2012;RAMOS et al, 2014;CARVALHO et al, 2016).…”

“…Argentina (ZUMÁRRAGA et al, 2013;GHAVIDEL et al, 2018 Estudos anteriores no Brasil também relataram esse espoligotipo (ROCHA et al, 2013;CARVALHO et al, 2016). Esse perfil já foi detectado na Ásia, Europa e na Américas, sendo um dos espoligotipos mais disperso pelo mundo (ANDRIEVSKAIA et al, 2018;GHAVIDEL et al, 2018).…”