Mycobacterium bovis identification by a molecular method from post-mortem inspected cattle obtained in abattoirs of Mato Grosso do Sul, Brazil

Araújo, Cristina Pires de; Leite, Clarice Q. F.; Prince, Karina Andrade de; Jorge, Klaudia dos Santos Gonçalves; Osório, Ana Luiza Alves Rosa

doi:10.1590/s0074-02762005000700013

Cited by 28 publications

(15 citation statements)

References 15 publications

(6 reference statements)

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“…All positive-culture specimens were also positive for m-PCR; and 10 (59%) samples that were negative by culturing yielded a positive result after m-PCR assay. It should be mentioned that the PCR was sensitive enough to detect M. bovis in a large proportion (59%) of those samples that failed to grow in culture, as also reported by Liebana et al (1995), Zanini et al (2001) and Araújo et al (2005). The efficiency of the culture method used as a first criterion for M. bovis identification is low, because of the small number of live bacilli present in some tissues.…”

Section: Detection Of Mycobacterium Bovis Dna In Bovine Tissues By M-pcrmentioning

confidence: 64%

Molecular Techniques for Identification of Species of the Mycobacterium tuberculosis Complex: The use of Multiplex PCR and an Adapted HPLC Method for Identification of Mycobacterium bovis and Diagnosis of Bovine Tuberculosis

Figueiredo¹,

Júnior²,

Furlanetto³

et al. 2012

Understanding Tuberculosis - Global Experiences and Innovative Approaches to the Diagnosis

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Section: Detection Of Mycobacterium Bovis Dna In Bovine Tissues By M-pcrmentioning

confidence: 64%

Molecular Techniques for Identification of Species of the Mycobacterium tuberculosis Complex: The use of Multiplex PCR and an Adapted HPLC Method for Identification of Mycobacterium bovis and Diagnosis of Bovine Tuberculosis

Figueiredo¹,

Júnior²,

Furlanetto³

et al. 2012

Understanding Tuberculosis - Global Experiences and Innovative Approaches to the Diagnosis

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“…and identified by biochemical testing and Ziehl-Neelsen (ZN), as M. bovis. In contrast (Araújo et al 2005) 72 samples were selected during slaugther inspection in abattoirs in the state of Mato Grosso do Sul, Brazil, obtained only 23.6% of samples showed colonies suggestive of mycobacteria that were confirmed to be acid-fast bacilli by ZN. In the present study, mycobacteria were isolated in 64.3% of the samples, including half of the evaluated retrofaryngeal lymphnodes samples and all prescapular lymphnodes (9), lungs (2), and liver (1) samples.…”

Section: Isolation and Identification Of Bovine Tuberculosis In A Bramentioning

confidence: 99%

Isolation and identification of bovine tuberculosis in a Brazilian herd (São Paulo)

Nassar

Miyashiro

Oliveira³

et al. 2007

Mem. Inst. Oswaldo Cruz

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Mycobacterium was verified in animals from a Brazilian dairy herd, a total of 42 samples from 30 cows were submitted to culture and the isolated strains were analyzed by two polymerase chain reaction (PCR), the first specific for species belonging to the Mycobacterium complex (MTBC) (Bouvet et al. 1993). From a clinical and epidemiological perspective, a rapid method to differentiate both species is important for timely diagnosis, effective treatment (due to the intrinsic resistance of M. bovis to pirazynamide) and epidemiology of TB (Shah et al. 2002).The bovine population in Brazil consists of about 180 million animals, with prevalence of TB ranging between 0.9 and 2.9% depending on the region and kind of production (Kantor & Ritacco 1994). Between 1989 and 1998, official notification data indicated a national prevalence of 1.3% of infected animals (Brasil 2005). Due to disgonic and very slow growth, the identification of M. bovis by conventional biochemical methods is cumbersome and timeconsuming. Polymerase chain reaction (PCR) is a sensitive and fast diagnostic tool that can be used to detect the agent in clinical samples in 48 h, but the presence of inhibitors in samples can interfere with its performance (Haddad et al. 2004, Singh et al. 2004, Brasil 2005.Among the several PCR systems developed for detection of species composing the M. tuberculosis complex (MTBC) (Wards et al. 1995, Brasil 2005, we focused on two. One was described by Niyaz Ahmed et al.(1999) and based on specific primers (NZ1 and NZ2) for the insertion sequence IS 1081, selectively present in organisms of the MTBC. A multiplex-PCR with primers JB and specific primers for the gene encoding for pyrazinimidase (pncA) was reported by Shah et al. (2002) to differentiate between M. bovis and M. tuberculosis on colonies in culture and this method could detect less than 20 pg of DNA, and this last reaction was the second applied reaction in the present work for differentiation between M. bovis and M. tuberculosis.We here present our work aiming at the detection of M. bovis mainly in retrofaryngeal lymphnode samples of bovines from a herd of Tatuí (São Paulo, Brazil) by microbiological culture, and MTBC and M. bovis specific PCR systems, and comparing the microbiological results with the positive or inconclusive results at single intradermal comparative tuberculin test (SICTT) presented by these animals.Thirty bovines from a Tatuí herd in São Paulo, Southeast Brazil and with positive (> 4 mm) or inconclusive (2 to 3.9 mm) response to SICTT were taken to sanitary slaughter. We collected 42 samples composed of the retrofaryngeal lymphnodes independent of the presence or not of suggestive lesions for TB while this was perfomed for prescapular lylmphnodes, lung, and liver only when presenting characteristic lesions of TB (Table). Samples were maintained in sterile bags at 4ºC for transport to the Laboratory of Tuberculosis of Instituto Biológico, São Paulo, Brazil until futher analysis.For culturing, samples were decontaminated by means of the Pet...

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“…Nesse estado, existem alguns estudos pontuais envolvendo o teste intradérmico, exame bacteriológico e molecular de lesões post mortem sugestivas de tuberculose que demonstram a presença da doença (Jorge 2001, Araújo et al 2005, Azambuja 2010). …”

Section: Introductionunclassified

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

et al. 2015

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Neste estudo, realizou-se genotipagem de isolados de Mycobacterium bovis, provenientes de amostras de tecidos de bovinos positivos no teste cervical comparativo (TCC) para tuberculose em Mato Grosso do Sul, por meio da técnica de spoligotyping. Tecidos de 13 bovinos positivos, oriundos de diferentes municípios do estado, foram cultivados em meio de Stonebrink. As colônias resultantes foram submetidas à coloração de Ziehl-Neelsen e todos os isolados apresentaram características tintoriais de BAAR. Os 13 isolados de BAAR foram identificados por PCR multiplex (mPCR). O gene hsp65 foi alvo para identificação de Mycobacterium spp, a sequência de inserção IS6110 foi alvo para identificação de complexo Mycobacterium tuberculosis (CMT) e a região rvd1rv2031c foi explorada para detecção de M. bovis. Os isolados micobacterianos foram genotipados pela técnica de spoligotyping. Dos 13 bovinos, sete tinham pelo menos uma lesão sugestiva de tuberculose em linfonodos retrofaríngeos, parotídeos e pulmonares ou no pulmão, e em seis não foram encontradas lesões visíveis sugestivas da doença. Na mPCR, 11/13 (84,6%) isolados foram positivos para Mycobacterium spp; 8/13 (61,5%) positivos para CMT e 7/13 (53,8%) positivos para M. bovis. Com base no spoligotyping, oito isolados de BAAR foram agrupados dentro de três diferentes agrupamentos de genótipos e uma amostra remanescente apresentou perfil único, sendo quatro isolados com padrão de espoligotipo SB0121, dois SB1145, dois SB0881 e um SB0140. A técnica de spoligotyping demonstrou que há diversidade genética entre os espoligotipos presentes no estado de Mato Grosso do Sul, embora predomine o perfil SB0121

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Mycobacterium bovis identification by a molecular method from post-mortem inspected cattle obtained in abattoirs of Mato Grosso do Sul, Brazil

Cited by 28 publications

References 15 publications

Molecular Techniques for Identification of Species of the Mycobacterium tuberculosis Complex: The use of Multiplex PCR and an Adapted HPLC Method for Identification of Mycobacterium bovis and Diagnosis of Bovine Tuberculosis

Molecular Techniques for Identification of Species of the Mycobacterium tuberculosis Complex: The use of Multiplex PCR and an Adapted HPLC Method for Identification of Mycobacterium bovis and Diagnosis of Bovine Tuberculosis

Isolation and identification of bovine tuberculosis in a Brazilian herd (São Paulo)

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

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