Identificação e genotipagem de Mycobacterium bovis em bovinos positivos no teste intradérmico para tuberculose em Mato Grosso do Sul

DE O. CAZOLA D; DOS S.G. JORGE K; ZUMÁRRAGA, MARTÍN JOSÉ; SOUZA-FILHO A.F; ARAÚJO FLÁBIO R; A.R. OSÓRIO A.L.

PESQUISA VETERINáRIA BRASILEIRA

REVISTA PESQUISA VETERINARIA BRASILEIRA

Año: 2015 vol. 35 p. 141 - 147

0100-736X

Spoligotyping was performed in the present study to genotype Mycobacterium bovis isolatesobtained from tissues of cattle that were positive in the comparative intradermal tuberculintest (CITT) in the state of Mato Grosso do Sul (Brazil). Tissue samples from 13 positivecattle from different municipalities of the state were cultured using a Stonebrink medium.The resulting colonies were subjected to Ziehl-Neelsen staining and all isolates exhibited thestaining characteristics of AFB. The 13 isolates of AFB were identified by means of a multiplexPCR (mPCR) assay. The hsp65 gene was targeted for the identification of Mycobacteriumspp., whereas the IS6110 insertion sequence was targeted for the identification of the Mycobacteriumtuberculosis complex (MTC) and the rvd1rv2031c region was explored for the detectionof Mycobacterium bovis. The spoligotyping assay was performed to genotype mycobacterialisolates. Of the 13 cattle, seven had at least one lesion suggestive of tuberculosis inthe retropharyngeal, parotid and lung lymph nodes or lung. The remaining six exhibited nolesions suggestive of the disease. In the mPCR, 11 of the 13 isolates (84.6%) were positive forMycobacterium spp., 8/13 (61.5%) were positive for the MTC and 7/13 (53.8%) were positivefor M. bovis. Based on the spoligotyping, eight isolates were grouped into three differentgroups of genotypes and one isolate exhibited an orphan type. Four isolates exhibited spoligotypepattern SB0121, while two isolates were associated with the pattern SB1145, anothertwo were associated with pattern SB0881 and one was associated with pattern SB0140.Spoligotyping confirmed the genetic diversity present among isolates found in the state ofMato Grosso do Sul. In addition, SB0121 was confirmed as the predominant profile.