Thin layer microcolony culture associated with PCR for early identification of Mycobacterium bovis

Rosário, Tatiana Reis do; Dib, Cristina Corsi; Roxo, Eliana; Pinheiro, Sônia Regina; Vasconcellos, Sílvio Arruda; Benites, Nilson Roberti

doi:10.1590/s1517-83822014005000038

Cited by 5 publications

(3 citation statements)

References 20 publications

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“…Slopes with loosened screw caps were incubated in air (IC-1) and in air containing 10% CO 2 (IC-2), and slopes closed with hydrophobic cotton were incubated in air. After 24 hours, the lids were tightened, and the cotton was burned and subsequently closed with a cork to create a higher CO 2 tension (IC-3) (ROSÁRIO et al, 2014). The incubation continued up to 90 days.…”

Section: Methodsmentioning

confidence: 99%

Influence of the incubation conditions on culture media to optimize primary isolation of Mycobacterium bovis

Ikuta

Morato

Souza

et al. 2016

SCA

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The isolation of Mycobacterium bovis is critical to a surveillance system for bovine tuberculosis based on detection of lesions in abattoirs. Thus, four solid culture media and three incubation conditions were investigated to elucidate which combination overcomes the others by assessing growth, time to the first appearance of colonies and their number. Ninety-seven samples of granulomatous lesions were submitted to the decontamination procedure by 1-hexadecylpyridinium chloride at 0.75% w/v, and inoculated on two egg-based media, Stonebrink's (ST) and Löwenstein-Jensen's with sodium pyruvate (LJp), and two agar-based media, tuberculosis blood agar (B83) and Middlebrook 7H11 medium (7H11). Each medium was incubated at 37°C for 90 days in three incubation conditions: in air, in air containing 10% carbon dioxide (CO 2 ), and in air in slopes closed with burned hydrophobic cotton and subsequently plugged with a cork to create a microaerophilic atmosphere. The colonies appeared faster and in higher number when incubated in air containing 10% CO 2 (p < 0.01), independent of media. B83 showed a faster growth and detected more isolates at 30 days of incubation, when compared to ST (0.0178), LJp (p < 0.0001) and 7H11 (p < 0.0001), though there was no difference between B83, ST and LJp at 60 and 90 days of incubation. 7H11 presented the lowest number of isolates (p < 0.0001) and a longer period for the appearance of the first colony (p < 0.001). According to our findings, the concomitant use of ST and B83 media incubated in air containing 10% CO 2 increases the isolation of M. bovis in a shorter period of time, which improves bovine tuberculosis diagnosis. Key words: Primary isolation. Bovine tuberculosis. Mycobacterium bovis. Culture media. Incubation conditions. ResumoO isolamento do Mycobacterium bovis é fundamental para um sistema de vigilância para tuberculose bovina baseado na detecção de lesões em abatedouro. Assim, quatro meios de cultura sólidos e três condições de incubação foram investigados para elucidar qual combinação supera as outras através da avaliação de crescimento, tempo para o aparecimento da primeira colônia e número de colônias. Noventa e sete amostras de lesões granulomatosas foram submetidas ao processo de descontaminação por cloreto de 1-hexadecilpiridínio a 0,75%, e inoculadas em dois meios a base de ovo, Stonebrink (ST) e Löwenstein-Jensen com piruvato de sódio (LJp), e dois meios a base de ágar, ágar sangue tuberculose (B83) e Middlebrook 7H11 (7H11). Cada meio foi incubado a 37°C por 90 dias, em três condições de incubação: em atmosfera normal, em atmosfera com acréscimo de 10% de dióxido de carbono (CO 2 ), e em atmosfera normal em tubos fechados com algodão hidrófobo queimado e subsequentemente fechado com rolha para criar uma atmosfera microaerófila. As colônias apareceram mais rapidamente e em maior número quando incubadas em atmosfera com 10% de CO 2 (p < 0,01), independente dos meios. As micobactérias cresceram em maior abundância e mais rapidamente no meio B83 aos 30 dias de i...

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Section: Methodsmentioning

confidence: 99%

Influence of the incubation conditions on culture media to optimize primary isolation of Mycobacterium bovis

Ikuta

Morato

Souza

et al. 2016

SCA

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“…Researchers have shown that the use of a Middlebrook 7H11 thin layer agar plate produces a marked difference in the speed of observation of the colonies of M. bovis and M. tuberculosis (MARCONDES et al, 2006). Apparently, in a relatively short period (between 12 to 25 days), it is possible to observe the growth of mycobacteria, which is an advantage over commonly used media, especially when it is associated with the use of polymerase chain reaction (PCR) for detection of microcolonies (ROSÁRIO et al, 2014).…”

Section: Diagnosismentioning

confidence: 99%

“…Tuberculosis in cattle is a public health concern and a potential threat to human health (DAVIES, 2006). Some authors believe that zoonotic tuberculosis is one of the many consequences of the adaptability of the Mycobacterium species to different hosts (ANAELOM et al, 2010), although this is controversial (ROTHSCHILD et al, 2001). …”

Section: Introductionmentioning

confidence: 99%

Tuberculosis diagnostic methods in buffaloes

et al. 2015

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The low productivity of buffalo herds and condemnation of carcasses in slaughterhouses due to tuberculosis lesions have resulted in increasing economic losses because these animals cannot be treated and must be destroyed by sanitary slaughter. Tuberculosis is a widely distributed zoonosis that affects the beef supply chain of the Brazilian agribusiness economically and socially. Like cattle, buffaloes are sensitive to Mycobacterium bovis, which is the main causative agent of zoonotic tuberculosis. Tuberculosis in buffaloes has been reported in several countries, including Brazil. In order to control and eradicate this disease among cattle and buffaloes in Brazil, the Ministry of Agriculture, Livestock, and Supply created the National Program for the Control and Eradication of Brucellosis and Tuberculosis with the main objective of finding a significant number of disease-free herds throughout the national territory using reliable methods. This review summarizes the main data on the history of occurrence of M. bovis in Brazilian herds and the diagnostic methods for the disease in buffaloes. Little information is available on buffalo tuberculosis. Due to the increasing population of buffaloes and their economic importance, more studies investigating the occurrence and identification of tuberculosis in this species are clearly needed. Key words: Mycobacterium bovis, isolation, zoonosis, bovine ResumoSão crescentes as perdas econômicas com baixa na produtividade dos rebanhos bubalinos e a condenação de carcaças em matadouros devido à tuberculose, uma vez que os animais não podem ser tratados e devem ser descartados em abates sanitários. A doença tem impacto econômico e social para a cadeia produtiva do agronegócio brasileiro, além de ser uma zoonose de ampla distribuição. Assim como os bovinos, o búfalo é sensível ao Mycobacterium bovis, principal agente da tuberculose zoonótica. A ocorrência da tuberculose nessa espécie tem sido relatada em diversos países, inclusive no Brasil. Para controle e erradicação dessa enfermidade entre os bovinos e bubalinos no Brasil, o Ministério da Agricultura Pecuária e Abastecimento (MAPA) instituiu o Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCEBT), cujo principal objetivo é, por meio de métodos confiáveis, obter significativo número de rebanhos livres dessas doenças, em todo o território nacional. Esta revisão reuniu os principais dados referentes ao histórico da ocorrência de M. bovis nos rebanhos do Brasil e aos métodos diagnósticos da doença em búfalos. Na literatura faltam dados relativos à tuberculose bubalina e com o incremento da criação desses animais e de sua importância econômica, notou-se evidente necessidade de estudos e pesquisas relativos à ocorrência e identificação da tuberculose nessa espécie. Palavras-chave: Mycobacterium bovis, isolamento, zoonose, bovídeos

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A systematic review on the distribution of Mycobacterium bovis infection among wildlife in the Americas

Domingos

Júnior

Lilenbaum

et al. 2019

Trop Anim Health Prod

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Thin layer microcolony culture associated with PCR for early identification of Mycobacterium bovis

Cited by 5 publications

References 20 publications

Influence of the incubation conditions on culture media to optimize primary isolation of Mycobacterium bovis

Influence of the incubation conditions on culture media to optimize primary isolation of Mycobacterium bovis

Tuberculosis diagnostic methods in buffaloes

A systematic review on the distribution of Mycobacterium bovis infection among wildlife in the Americas

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