Mastitis in cattle due to Mycobacteroides abscessus - case report / [Mastite bovina por Mycobacteroides abscessus - relato de caso]

Mastitis is considered the main disease that affects dairy cattle worldwide, and it is caused mainly by Staphylococcus aureus and environmental Streptococcus spp. Eventually, nonconventional pathogens, as rapidly growing mycobacteria (RGM), may also cause chronic mastitis, which will not be responsive to antibiotic treatments. Diagnosis of mastitis caused by RGM is a difficult task, and most of time this agent may be misdiagnosed. Here we describe a case of clinical mastitis caused by the RGM Mycobacteroides abscessus in a cow from Southern Brazil, confirmed by microbiological and molecular characterization. Our results reinforce the necessity of a detailed laboratorial identification of the agent and to include this agent in differential diagnosis of chronical clinical mastitis nonresponsive to treatment.

A mastite é considerada uma das principais doenças que afetam rebanhos leiteiros ao redor do mundo e é causada principalmente por Staphylococcus aureus e Streptococcus spp. ambientais. Eventualmente, patógenos não convencionais, como Mycobacterium de crescimento rápido (RGM), podem também causar mastite crônica, a qual não respondera ao tratamento com antimicrobianos. O diagnóstico de mastite causada por RGM é uma tarefa difícil, e a maioria das vezes esse agente pode ser subdiagnosticado. Neste relato, descreveu-se um caso de mastite clínica em uma vaca, no sul do Brasil, causada por RGM Mycobacteroides abscessus, confirmado por caracterização microbiológica e molecular. Os resultados reforçam a necessidade de um diagnóstico laboratorial detalhado para a identificação do agente e a inclusão deste como diagnóstico diferencial no reconhecimento de mastite crônica não responsiva ao tratamento.

Tuberculose bovina: impactos para a pecuária e riscos para a Saúde Pública / Bovine tuberculosis: impacts for livestock and risks for Public Health / Tuberculosis bovina: impactos para el ganado y riesgos para la Salud Pública

A tuberculose bovina está entre as principais zoonoses detectadas pelo serviço de inspeção durante o exame post-mortem das carcaças bovinas, sendo responsável por gerar consideráveis prejuízos econômicos para a pecuária e riscos para a saúde pública. Tem como agente etiológico o microrganismo Mycobacterium bovis, o qual possui tropismo pela espécie bovina, porém pode acometer outras espécies, incluindo a humana. Estima-se que nos países desenvolvidos, 1 a 2% dos casos de tuberculose em humanos seja causada por M. bovis, enquanto nos subdesenvolvidos, esta estimativa é de 10 a 20%, podendo ser ainda maior. Já, para a pecuária, as perdas são decorrentes da redução de 10 a 25% na eficiência produtiva, no ganho de peso, pela necessidade de sacrifício dos animais positivos e das condenações totais ou parciais das carcaças oriundas de animais infectados. O presente estudo teve por objetivo abordar os impactos da tuberculose bovina para a pecuária e sua importância para a saúde pública.

Bovine tuberculosis is among the main zoonosis detected by the inspection service during the postmortem examination of bovine carcasses, and is responsible for generating considerable economic damage to livestock and risks to public health. Its etiological agent is the microorganism Mycobacterium bovis, which has tropism for the bovine species, but can affect other species, including the human one. It is estimated that in developed countries 1 to 2% of tuberculosis cases in humans are caused by M. bovis, while in underdeveloped countries, this estimate is 10 to 20% and may be even higher. For livestock, losses are due to a 10 to 25% reduction in productive efficiency, weight gain, the need to sacrifice positive animals and total or partial condemnation of carcasses from infected animals. This study aimed to address the impacts of bovine tuberculosis on livestock and its importance to public health.

La tuberculosis bovina se encuentra entre las principales zoonosis detectadas por el servicio de inspección durante el examen post mortem de los cadáveres bovinos, y es responsable de generar considerables daños económicos para el ganado y riesgos para la salud pública. Su agente etiológico es el microorganismo Mycobacterium bovis, que tiene tropismo para las especies bovinas, pero puede afectar a otras especies, incluida la humana. Se estima que en los países desarrollados 1 a 2% de los casos de tuberculosis en humanos son causados por M. bovis, mientras que en los países subdesarrollados, esta estimación es del 10 al 20% y puede ser aún mayor. Para el ganado, las pérdidas se deben a una reducción del 10 al 25% en la eficiencia productiva, el aumento de peso, la necesidad de sacrificar animales positivos y la condena total o parcial de los cadáveres de animales infectados. Este estudio tuvo como objetivo abordar los impactos de la tuberculosis bovina en el ganado y su importancia para la salud pública.

Tuberculose bovina: impactos para a pecuária e riscos para a Saúde Pública / Bovine tuberculosis: impacts for livestock and risks for Public Health / Tuberculosis bovina: impactos para el ganado y riesgos para la Salud Pública

A tuberculose bovina está entre as principais zoonoses detectadas pelo serviço de inspeção durante o exame post-mortem das carcaças bovinas, sendo responsável por gerar consideráveis prejuízos econômicos para a pecuária e riscos para a saúde pública. Tem como agente etiológico o microrganismo Mycobacterium bovis, o qual possui tropismo pela espécie bovina, porém pode acometer outras espécies, incluindo a humana. Estima-se que nos países desenvolvidos, 1 a 2% dos casos de tuberculose em humanos seja causada por M. bovis, enquanto nos subdesenvolvidos, esta estimativa é de 10 a 20%, podendo ser ainda maior. Já, para a pecuária, as perdas são decorrentes da redução de 10 a 25% na eficiência produtiva, no ganho de peso, pela necessidade de sacrifício dos animais positivos e das condenações totais ou parciais das carcaças oriundas de animais infectados. O presente estudo teve por objetivo abordar os impactos da tuberculose bovina para a pecuária e sua importância para a saúde pública.(AU)

Bovine tuberculosis is among the main zoonosis detected by the inspection service during the postmortem examination of bovine carcasses, and is responsible for generating considerable economic damage to livestock and risks to public health. Its etiological agent is the microorganism Mycobacterium bovis, which has tropism for the bovine species, but can affect other species, including the human one. It is estimated that in developed countries 1 to 2% of tuberculosis cases in humans are caused by M. bovis, while in underdeveloped countries, this estimate is 10 to 20% and may be even higher. For livestock, losses are due to a 10 to 25% reduction in productive efficiency, weight gain, the need to sacrifice positive animals and total or partial condemnation of carcasses from infected animals. This study aimed to address the impacts of bovine tuberculosis on livestock and its importance to public health.(AU)

La tuberculosis bovina se encuentra entre las principales zoonosis detectadas por el servicio de inspección durante el examen post mortem de los cadáveres bovinos, y es responsable de generar considerables daños económicos para el ganado y riesgos para la salud pública. Su agente etiológico es el microorganismo Mycobacterium bovis, que tiene tropismo para las especies bovinas, pero puede afectar a otras especies, incluida la humana. Se estima que en los países desarrollados 1 a 2% de los casos de tuberculosis en humanos son causados por M. bovis, mientras que en los países subdesarrollados, esta estimación es del 10 al 20% y puede ser aún mayor. Para el ganado, las pérdidas se deben a una reducción del 10 al 25% en la eficiencia productiva, el aumento de peso, la necesidad de sacrificar animales positivos y la condena total o parcial de los cadáveres de animales infectados. Este estudio tuvo como objetivo abordar los impactos de la tuberculosis bovina en el ganado y su importancia para la salud pública.(AU)

Mycobacteria in Minas cheese commercialized in open fairs in São Paulo, Brazil / Mycobacteria em queijos tipo Minas comercializados em feiras-livres de São Paulo, Brasil

Mycobacterium bovis is the causative agent of bovine tuberculosis, a disease that affects dairy herds throughout the Brazilian territory, constituting a neglected zoonosis transmitted by raw milk and its derivatives. In this study, we evaluated the presence of M. bovis and other mycobacteria in Minas cheese obtained from open fairs in the city of São Paulo between 2012 and 2013. Samples (n = 133) were decontaminated using hexa-cetylpyridinium chloride and seeded on StonebrinkLeslie medium. The isolates were submitted to molecular identification by TB Multiplex PCR targeting the 16S rRNA gene and amplicon nucleotide sequencing. From 16 cheese samples (12%), we obtained 26 putative colonies of Mycobacterium spp., none of which belonged to any of the Mycobacterium tuberculosis, Mycobacterium avium, or Mycobacterium intracellulare complexes. Phylogenetic analysis showed that sample sequences were grouped in a clade that includes only non-tuberculous mycobacteria with proximity to sequences obtained from Mycobacterium novocastrense (3 sequences), Mycobacterium holsaticum (1 sequence), andMycobacterium elephantis (2 sequences). Although no epidemiological evidence was found regarding the importance of oral transmission of mycobacteria in healthy people, their importance in the immunosuppressed population remains uncertain.(AU)

Mycobacterium bovis é o agente da tuberculose bovina, doença que acomete o rebanho em todo território brasileiro e é uma negligenciada zoonose transmitida pelo leite e seus derivados. Este trabalho avaliou a presença de M. bovis e outras micobactérias, em queijo minas meia-cura, obtidos em feiras-livres na cidade de São Paulo, entre os anos de 2012 e 2013. As amostras (n = 133) foram descontaminadas pelo método HPC (hexa-cetyl-pyridinium chloride) e semeadas em meio Stonebrink Leslie. Os isolados foram submetidos à identificação molecular por PCR TB multiplex, pesquisando-se o gene 16S rRNA, e ao sequenciamento nucleotídico. Dezesseis amostras (12%) possuiam 26 colônias sugestivas de Mycobacterium spp., mas nenhuma delas pertencia aos complexos Mycobacterium tuberculosis, Mycobacterium avium e Mycobacterium intracellulare. A análise filogenética mostrou que todas as amostras estavam agrupadas em clados que incluem apenas micobactérias não tuberculosas (MNT), sendo que algumas possuiam proximidade com sequências obtidas de Mycobacterium novocastrense (3 sequências), Mycobacterium hosaticum (1 sequência) e Mycobacterium elephantis (2 sequências). Embora no momento não haja evidência epidemiológica da importância da transmissão oral das micobactérias pra indivíduos saudáveis, sua importância na população imunossuprimida ainda é incerta.(AU)

Mycobacteria in Minas cheese commercialized in open fairs in São Paulo, Brazil / Mycobacteria em queijos tipo Minas comercializados em feiras-livres de São Paulo, Brasil

Mycobacterium bovis is the causative agent of bovine tuberculosis, a disease that affects dairy herds throughout the Brazilian territory, constituting a neglected zoonosis transmitted by raw milk and its derivatives. In this study, we evaluated the presence of M. bovis and other mycobacteria in Minas cheese obtained from open fairs in the city of São Paulo between 2012 and 2013. Samples (n = 133) were decontaminated using hexa-cetylpyridinium chloride and seeded on Stonebrink­Leslie medium. The isolates were submitted to molecular identification by TB Multiplex PCR targeting the 16S rRNA gene and amplicon nucleotide sequencing. From 16 cheese samples (12%), we obtained 26 putative colonies of Mycobacterium spp, none of which belonged to any of the Mycobacterium tuberculosis, Mycobacterium avium, or Mycobacterium intracellulare complexes. Phylogenetic analysis showed that sample sequences were grouped in a clade that includes only non-tuberculous mycobacteria with proximity to sequences obtained from Mycobacterium novocastrense (3 sequences), Mycobacterium holsaticum (1 sequence), and Mycobacterium elephantis (2 sequences). Although no epidemiological evidence was found regarding the importance of oral transmission of mycobacteria in healthy people, their importance in the immunosuppressed population remains uncertain.(AU)

Mycobacterium bovis é o agente da tuberculose bovina, doença que acomete o rebanho em todo território brasileiro e é uma negligenciada zoonose transmitida pelo leite e seus derivados. Este trabalho avaliou a presença de M. bovise outras micobactérias, em queijo minas meia-cura, obtidos em feiras-livres na cidade de São Paulo, entre os anos de 2012 e 2013. As amostras (n = 133) foram descontaminadas pelo método HPC (hexa-cetyl-pyridinium chloride) e semeadas em meio Stonebrink Leslie. Os isolados foram submetidos à identificação molecular por PCR TB multiplex, pesquisando-se o gene 16S rRNA, e ao sequenciamento nucleotídico. Dezesseis amostras (12%) possuiam 26 colônias sugestivas de Mycobacterium spp, mas nenhuma delas pertencia aos complexos Mycobacterium tuberculosis, Mycobacterium avium e Mycobacterium intracellulare. A análise filogenética mostrou que todas as amostras estavam agrupadas em clados que incluem apenas micobactérias não tuberculosas (MNT), sendo que algumas possuiam proximidade com sequências obtidas de Mycobacterium novocastrense (3 sequências), Mycobacterium hosaticum(1 sequência) e Mycobacterium elephantis (2 sequências). Embora no momento não haja evidência epidemiológica da importância da transmissão oral das micobactérias pra indivíduos saudáveis, sua importância na população imunossuprimida ainda é incerta.(AU)

Mycobacteria in Minas cheese commercialized in open fairs in São Paulo, Brazil / Mycobacteria em queijos tipo Minas comercializados em feiras-livres de São Paulo, Brasil

Mycobacterium bovis is the causative agent of bovine tuberculosis, a disease that affects dairy herds throughout the Brazilian territory, constituting a neglected zoonosis transmitted by raw milk and its derivatives. In this study, we evaluated the presence of M. bovis and other mycobacteria in Minas cheese obtained from open fairs in the city of São Paulo between 2012 and 2013. Samples (n = 133) were decontaminated using hexa-cetylpyridinium chloride and seeded on StonebrinkLeslie medium. The isolates were submitted to molecular identification by TB Multiplex PCR targeting the 16S rRNA gene and amplicon nucleotide sequencing. From 16 cheese samples (12%), we obtained 26 putative colonies of Mycobacterium spp., none of which belonged to any of the Mycobacterium tuberculosis, Mycobacterium avium, or Mycobacterium intracellulare complexes. Phylogenetic analysis showed that sample sequences were grouped in a clade that includes only non-tuberculous mycobacteria with proximity to sequences obtained from Mycobacterium novocastrense (3 sequences), Mycobacterium holsaticum (1 sequence), andMycobacterium elephantis (2 sequences). Although no epidemiological evidence was found regarding the importance of oral transmission of mycobacteria in healthy people, their importance in the immunosuppressed population remains uncertain.

Mycobacterium bovis é o agente da tuberculose bovina, doença que acomete o rebanho em todo território brasileiro e é uma negligenciada zoonose transmitida pelo leite e seus derivados. Este trabalho avaliou a presença de M. bovis e outras micobactérias, em queijo minas meia-cura, obtidos em feiras-livres na cidade de São Paulo, entre os anos de 2012 e 2013. As amostras (n = 133) foram descontaminadas pelo método HPC (hexa-cetyl-pyridinium chloride) e semeadas em meio Stonebrink Leslie. Os isolados foram submetidos à identificação molecular por PCR TB multiplex, pesquisando-se o gene 16S rRNA, e ao sequenciamento nucleotídico. Dezesseis amostras (12%) possuiam 26 colônias sugestivas de Mycobacterium spp., mas nenhuma delas pertencia aos complexos Mycobacterium tuberculosis, Mycobacterium avium e Mycobacterium intracellulare. A análise filogenética mostrou que todas as amostras estavam agrupadas em clados que incluem apenas micobactérias não tuberculosas (MNT), sendo que algumas possuiam proximidade com sequências obtidas de Mycobacterium novocastrense (3 sequências), Mycobacterium hosaticum (1 sequência) e Mycobacterium elephantis (2 sequências). Embora no momento não haja evidência epidemiológica da importância da transmissão oral das micobactérias pra indivíduos saudáveis, sua importância na população imunossuprimida ainda é incerta.

Isolation and identification of Mycobacterium bovis in bovines with positive reaction to the tuberculin test in the state of Paraíba, northeast Brazil / Isolamento e identificação de Mycobacterium bovis em bovinos positivos no teste de tuberculinização no estado da Paraíba, nordeste do Brasil

In areas where human tuberculosis and bovine tuberculosis coexist, differentiation between M. bovis and M. tuberculosis is important for monitoring the spread of M. bovis among cattle and from cattle to humans. The objective of this study was to isolate and identify M. bovis in bovines with positive diagnosis identified on tuberculin test in the State of Paraíba, Northeastern Brazil. Thirty-two bovines that tested positive in the comparative tuberculin test were used, from which samples of any organ with lesions suggestive of tuberculosis were collected, as well as lymph nodes, when no gross lesions were observed. Samples were submitted to histopathological exam, mycobacterial culture, Ziehl-Neelsen staining and molecular diagnosis. Twenty-one (65.6%) animals presented lesions suggestive of tuberculosis. As to body region 77.7% of lesions were found in the thoracic cavity, 12.4% in the head and 9.9% in the abdominal cavity. Among 55 samples submitted to mycobacterial culture, mycobacteria were isolated in 31 (56.4%), being 13 (41.9%) identified as M. bovis and 18 (58.1%) as Mycobacterium spp. Conclusion is that isolation and identification of M. bovis and Mycobacterium spp. in cattle suggests that humans are exposed to the risk of infection. This reinforces the need for intensification and optimization of prevention and control measures foreseen in the Brazilian National Program for the Control and Eradication of Bovine Brucellosis and Tuberculosis. Mycobacteria isolation and identification surveys are, therefore, encouraged in other Northeastern states.(AU)

Em áreas onde a tuberculose humana e a tuberculose bovina coexistem, a diferenciação entre M. bovis e M. tuberculosis é importante para monitorar a disseminação de M. bovis entre bovinos e destes para os seres humanos. Objetivou-se neste estudo isolar e identificar M. bovis em bovinos com diagnóstico positivo pelo teste de tuberculinização no estado da Paraíba, nordeste do Brasil. Foram submetidos 32 bovinos positivos ao teste de tuberculinização comparativa, dos quais foram colhidas amostras de qualquer órgão com lesões sugestivas de tuberculose, e, nos casos em que não foram observadas lesões sugestivas, foram colhidas amostras de linfonodos. As amostras foram submetidas a exame histopatológico, cultivo micobacteriológico, coloração de Ziehl-Neelsen e diagnóstico molecular. Apresentaram lesões sugestivas de tuberculose 21 animais (65,6%). Com relação à distribuição das lesões de acordo com a região corporal, 77,7% localizavam-se na cavidade torácica, 12,4% na cabeça e 9,9% na cavidade abdominal. De 55 amostras submetidas ao cultivo de micobactérias, em 31 (56,4%) foram isoladas micobactérias, sendo que em 13 (41,9%) foi identificado M. bovis, e nas 18 restantes (58,1%) foi identificado Mycobacterium spp. Conclui-se que o isolamento e a identificação de M. bovis e Mycobacterium spp. em bovinos indicam que os seres humanos estão expostos ao risco de infecção. Isso reforça a necessidade de intensificação e otimização de medidas de prevenção e controle previstas no Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose Bovina. Sugere-se a realização de estudos de isolamento e identificação de micobactérias em outros estados do Nordeste.(AU)

Isolation and identification of Mycobacterium bovis in bovines with positive reaction to the tuberculin test in the state of Paraíba, northeast Brazil / Isolamento e identificação de Mycobacterium bovis em bovinos positivos no teste de tuberculinização no estado da Paraíba, nordeste do Brasil

In areas where human tuberculosis and bovine tuberculosis coexist, differentiation between M. bovis and M. tuberculosis is important for monitoring the spread of M. bovis among cattle and from cattle to humans. The objective of this study was to isolate and identify M. bovis in bovines with positive diagnosis identified on tuberculin test in the State of Paraíba, Northeastern Brazil. Thirty-two bovines that tested positive in the comparative tuberculin test were used, from which samples of any organ with lesions suggestive of tuberculosis were collected, as well as lymph nodes, when no gross lesions were observed. Samples were submitted to histopathological exam, mycobacterial culture, Ziehl-Neelsen staining and molecular diagnosis. Twenty-one (65.6%) animals presented lesions suggestive of tuberculosis. As to body region 77.7% of lesions were found in the thoracic cavity, 12.4% in the head and 9.9% in the abdominal cavity. Among 55 samples submitted to mycobacterial culture, mycobacteria were isolated in 31 (56.4%), being 13 (41.9%) identified as M. bovis and 18 (58.1%) as Mycobacterium spp. Conclusion is that isolation and identification of M. bovis and Mycobacterium spp. in cattle suggests that humans are exposed to the risk of infection. This reinforces the need for intensification and optimization of prevention and control measures foreseen in the Brazilian National Program for the Control and Eradication of Bovine Brucellosis and Tuberculosis. Mycobacteria isolation and identification surveys are, therefore, encouraged in other Northeastern states.(AU)

Em áreas onde a tuberculose humana e a tuberculose bovina coexistem, a diferenciação entre M. bovis e M. tuberculosis é importante para monitorar a disseminação de M. bovis entre bovinos e destes para os seres humanos. Objetivou-se neste estudo isolar e identificar M. bovis em bovinos com diagnóstico positivo pelo teste de tuberculinização no estado da Paraíba, nordeste do Brasil. Foram submetidos 32 bovinos positivos ao teste de tuberculinização comparativa, dos quais foram colhidas amostras de qualquer órgão com lesões sugestivas de tuberculose, e, nos casos em que não foram observadas lesões sugestivas, foram colhidas amostras de linfonodos. As amostras foram submetidas a exame histopatológico, cultivo micobacteriológico, coloração de Ziehl-Neelsen e diagnóstico molecular. Apresentaram lesões sugestivas de tuberculose 21 animais (65,6%). Com relação à distribuição das lesões de acordo com a região corporal, 77,7% localizavam-se na cavidade torácica, 12,4% na cabeça e 9,9% na cavidade abdominal. De 55 amostras submetidas ao cultivo de micobactérias, em 31 (56,4%) foram isoladas micobactérias, sendo que em 13 (41,9%) foi identificado M. bovis, e nas 18 restantes (58,1%) foi identificado Mycobacterium spp. Conclui-se que o isolamento e a identificação de M. bovis e Mycobacterium spp. em bovinos indicam que os seres humanos estão expostos ao risco de infecção. Isso reforça a necessidade de intensificação e otimização de medidas de prevenção e controle previstas no Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose Bovina. Sugere-se a realização de estudos de isolamento e identificação de micobactérias em outros estados do Nordeste.(AU)

Use of green fluorescent protein labeled non-tuberculous mycobacteria to evaluate the activity quaternary ammonium compound disinfectants and antibiotics

Although infections with NonTuberculous Mycobacteria have become less common in AIDS patients, they are important opportunistic infections after surgical procedures, likely because they are ubiquitous and not efficiently killed by many commonly used disinfectants. In Venezuela there have recently been many non-tuberculous mycobacteria soft tissue infections after minor surgical procedures, some apparently related to the use of a commercial disinfectant based on a Quaternary Ammonium Compound. We studied the activity of this and other quaternary ammonium compounds on different non-tuberculous mycobacteria by transforming the mycobacteria with a dnaA-gfp fusion and then monitoring fluorescence to gauge the capacity of different quaternary ammonium compounds to inhibit bacterial growth. The minimum inhibitory concentration varied for the different quaternary ammonium compounds, but M. chelonae and M. abscessus were consistently more resistant than M. smegmatis, and M. terrae more resistant than M. bovis BCG.(AU)

Identification of Mycobacterium smegmatis in bovine mastitis

Background: Rapidly growing mycobacteria (RGM) are ubiquitous in the environment, can be isolated from soil and water, and demonstrate visible growth on culture media within seven days. Mycobacterium smegmatis is an acid-alcohol fast bacterium, which belong to RGM group. The diagnosis of M. smegmatis infections may be quite difficult by conventional methods; therefore, biochemistry associated to nucleic acid-based approaches provided fast and accurate identification. Although this specie may be associated to animals and humans infections, there is few cases description. Nontuberculous mycobacterial bovine mastitis is uncommon, and bovine mastitis by M. smegmatis has been reported but non-confirmed case once in the past. This paper reports M. smegmatis recovered from a cattle with relapsing pyogranulomatous mastitis. Case: Milk samples from an adult Holstein cow showing relapsing pyogranulomatous mastitis history and by pronounced glandular hardening were cultivated and analyzed accordingly to standard milk cultivation protocols. The animal had been subject to several intramammary and parenteral antibiotic therapies protocols without adequate response. After 48 h incubation, a slow and sparse growth of slightly pigmented, shiny and smooth colonies was observed on the blood agar plate. The bacterium isolated was named as strain 55/08. The morphological and biochemical profile [...]

Identification of Mycobacterium smegmatis in bovine mastitis

Background: Rapidly growing mycobacteria (RGM) are ubiquitous in the environment, can be isolated from soil and water, and demonstrate visible growth on culture media within seven days. Mycobacterium smegmatis is an acid-alcohol fast bacterium, which belong to RGM group. The diagnosis of M. smegmatis infections may be quite difficult by conventional methods; therefore, biochemistry associated to nucleic acid-based approaches provided fast and accurate identification. Although this specie may be associated to animals and humans infections, there is few cases description. Nontuberculous mycobacterial bovine mastitis is uncommon, and bovine mastitis by M. smegmatis has been reported but non-confirmed case once in the past. This paper reports M. smegmatis recovered from a cattle with relapsing pyogranulomatous mastitis. Case: Milk samples from an adult Holstein cow showing relapsing pyogranulomatous mastitis history and by pronounced glandular hardening were cultivated and analyzed accordingly to standard milk cultivation protocols. The animal had been subject to several intramammary and parenteral antibiotic therapies protocols without adequate response. After 48 h incubation, a slow and sparse growth of slightly pigmented, shiny and smooth colonies was observed on the blood agar plate. The bacterium isolated was named as strain 55/08. The morphological and biochemical profile [...](AU)

Effect of pasteurization on the decay of Mycobacterium bovis in milk cream / Efeito da pasteurização no decaimento de Mycobacterium bovis em creme de leite

Milk cream must be pasteurized in order to be sold in Brazil. However, there are no specific legal requirements for this product, and producers set their own pasteurization parameters using the ones approved for milk as a reference. Considering that fat protects bacteria from heat, that no thermal inactivation studies have been performed on Mycobacterium bovis present in cream, and that bovine tuberculosis is endemic in Brazil, the aim of this study was to evaluate the inactivation of M. bovis in milk cream subjected to commercial parameters of pasteurization. Milk cream samples were contaminated and pasteurized in a water bath at 75, 80, 85, and 90C for 5 and 15 s. M. bovis cells were plated onto Stonebrink-Leslie medium, incubated at 36C for 45 days, and quantified; the result was expressed in log CFU mL-1. The fat content of the samples ranged from 34% to 37% and the average initial load of M. bovis was 8.0 Log CFU mL-1. The average decay of the M. bovis populations was 4.0, 4.3, 4.9 and 6.7 log CFU mL-1 when the cream was incubated for 15 sec at 75, 80, 85 and 90C, respectively, showing that the efficiency of the heat treatment was improved by increasing the temperature of the process. Given the lipophilic nature of M. bovis, the cream should be subjected to more intense parameters of pasteurization than those applied to milk.(AU)

A pasteurização do creme de leite é obrigatória no Brasil, mas não há parâmetros legais específicos para esse produto, de forma que as empresas estabelecem seus próprios parâmetros tendo como referência mínima os aprovados para a pasteurização do leite. Assim, considerando que a gordura tem efeito termo protetor para as bactérias, que não há estudos de inativação térmica do Mycobacterium bovis em creme de leite e que a tuberculose bovina é endêmica no Brasil, o objetivo deste estudo foi avaliar a inativação de M. bovis em creme de leite, submetido aos parâmetros comerciais de pasteurização. Amostras de creme de leite foram contaminadas e pasteurizadas em Banho-Maria a 75C, 80C, 85C e 90C, por até 15 s. O M. bovis foi quantificado por semeadura em meio Stonebrink-Leslie, incubado a 36ºC por 45 dias, e o resultado expresso em Log UFC mL-1. O teor de gordura das amostras variou de 34 a 37% e a carga inicial média de M. bovis foi de 8,0 Log UFC mL-1. O decaimento médio da população de M. bovis foi de 4,0, 4,3, 4,9 e 6,7 Log UFC mL-1, quando expostos respectivamente às temperaturas estudadas por 15 s, demonstrando que a eficácia do tratamento térmico melhorou com o aumento da temperatura do processo. Devido ao caráter lipofílico do M. bovis, o creme de leite deveria ser submetido a parâmetros de pasteurização mais intensos do que aqueles aplicados ao leite.(AU)

Effect of pasteurization on the decay of Mycobacterium bovis in milk cream / Efeito da pasteurização no decaimento de Mycobacterium bovis em creme de leite

Milk cream must be pasteurized in order to be sold in Brazil. However, there are no specific legal requirements for this product, and producers set their own pasteurization parameters using the ones approved for milk as a reference. Considering that fat protects bacteria from heat, that no thermal inactivation studies have been performed on Mycobacterium bovis present in cream, and that bovine tuberculosis is endemic in Brazil, the aim of this study was to evaluate the inactivation of M. bovis in milk cream subjected to commercial parameters of pasteurization. Milk cream samples were contaminated and pasteurized in a water bath at 75, 80, 85, and 90C for 5 and 15 s. M. bovis cells were plated onto Stonebrink-Leslie medium, incubated at 36C for 45 days, and quantified; the result was expressed in log CFU mL-1. The fat content of the samples ranged from 34% to 37% and the average initial load of M. bovis was 8.0 Log CFU mL-1. The average decay of the M. bovis populations was 4.0, 4.3, 4.9 and 6.7 log CFU mL-1 when the cream was incubated for 15 sec at 75, 80, 85 and 90C, respectively, showing that the efficiency of the heat treatment was improved by increasing the temperature of the process. Given the lipophilic nature of M. bovis, the cream should be subjected to more intense parameters of pasteurization than those applied to milk.

A pasteurização do creme de leite é obrigatória no Brasil, mas não há parâmetros legais específicos para esse produto, de forma que as empresas estabelecem seus próprios parâmetros tendo como referência mínima os aprovados para a pasteurização do leite. Assim, considerando que a gordura tem efeito termo protetor para as bactérias, que não há estudos de inativação térmica do Mycobacterium bovis em creme de leite e que a tuberculose bovina é endêmica no Brasil, o objetivo deste estudo foi avaliar a inativação de M. bovis em creme de leite, submetido aos parâmetros comerciais de pasteurização. Amostras de creme de leite foram contaminadas e pasteurizadas em Banho-Maria a 75C, 80C, 85C e 90C, por até 15 s. O M. bovis foi quantificado por semeadura em meio Stonebrink-Leslie, incubado a 36ºC por 45 dias, e o resultado expresso em Log UFC mL-1. O teor de gordura das amostras variou de 34 a 37% e a carga inicial média de M. bovis foi de 8,0 Log UFC mL-1. O decaimento médio da população de M. bovis foi de 4,0, 4,3, 4,9 e 6,7 Log UFC mL-1, quando expostos respectivamente às temperaturas estudadas por 15 s, demonstrando que a eficácia do tratamento térmico melhorou com o aumento da temperatura do processo. Devido ao caráter lipofílico do M. bovis, o creme de leite deveria ser submetido a parâmetros de pasteurização mais intensos do que aqueles aplicados ao leite.

Decline in Mycobacterium bovis and Brucella abortus populations during the maturation of experimentally contaminated parmesan-type cheese / Decaimento do Mycobacterium bovis e da Brucella abortus durante a cura de queijo tipo parmesão experimentalmente contaminado

Brazilian legislation allows the manufacture of raw milk cheese with a maturation exceeding 60 days at room temperature above 5C, but there is a lack of solid scientific evidence on the efficacy of this maturation process in inactivating important pathogens that may be present in milk, such as Mycobacterium bovis and Brucella abortus. Thus, the objectives of this study were to produce parmesan-type cheese experimentally contaminated with M. bovis and B. abortus and evaluate the survival of these pathogens along 2-month maturation. Parmesan-type cheese was manufactured in the laboratory using whole pasteurized milk with or without inoculation with M. bovis (SB1033) or B. abortus (1119-3) and matured at 18C for up to 63 days. M. bovis was inoculated in Stonebrink-Leslie medium supplemented with antibiotics and incubated at 37C for 45 days, and B. abortus was incubated in Farrel medium at 36C for 3 days. The average D18C value, weighted by variance, was 37.5 ± 5.3 days for M. bovis and 5.9 ± 0.7 days for B. abortus. The average physicochemical parameters in the cheese at the end of the study were as follows: pH = 4.89, water activity = 0.976, and moisture percentage = 43.1%. The pH might have contributed to the reduction in the population of B. abortus but seems not to have influenced the population of M. bovis. We conclude that the duration of the maturation process influences the size of the surviving populations of M. bovis and B. abortus, and that the shortening of the maturation duration might not ensure a decline in pathogen levels to safe levels. Thus, complementary studies considering the effect of several other technological aspects on the survival of these pathogens are required, including the effect of the lactic acid bacterial population, salt content, and temperature of maturation.(AU)

A legislação brasileira permite a fabricação queijos de leite cru com maturação superior a 60 dias em temperatura acima de 5C, mas falta evidência científica sólida sobre a eficácia do processo de maturação na inativação de importantes patógenos que podem estar presentes no leite, como o Mycobacterium bovis e a Brucella abortus. Assim, os objetivos desse estudo foram produzir o queijo tipo parmesão experimentalmente contaminado com M. bovis e B. abortus e avaliar a sobrevivência desses patógenos ao longo de 2 meses de maturação. Queijo do tipo parmesão foi fabricado em laboratório com leite pasteurizado integral com ou sem inoculo de M. bovis (SB1033) ou B. abortus (1119-3) e maturado a 18C por até 63 dias. M. bovis foi semeado em meio Stonebrink-Leslie, acrescido de antibióticos, e incubado a 37C por 45 dias e a B. abortus foi semeada em meio Farrel a 36C por 3 dias. O valor D18C médio, ponderado pela variância, foi de 37,5 ± 5,3 dias para o M. bovis e de 5,9 ± 0,7 dias para a B. abortus. Os parâmetros físico-químicos médios do queijo no final do estudo foram como se segue: pH = 4,89, atividade de água = 0,976 e umidade percentual = 43,1%. O pH pode ter contribuído para redução da população de B. abortus mas parece não ter influenciado a população de M. bovis. Concluiu-se que a duração do processo de maturação influencia o tamanho da população sobrevivente de M. bovis e de B. abortus e que o encurtamento do período de maturação pode não garantir um declínio desses patógenos para níveis seguros. Assim, estudos complementares que considerem o efeito de vários outros aspectos tecnológicos na sobrevivência desses patógenos são necessários, incluindo o efeito da população de bactérias ácido láticas, teor de sal e temperatura de maturação.(AU)

Decline in Mycobacterium bovis and Brucella abortus populations during the maturation of experimentally contaminated parmesan-type cheese / Decaimento do Mycobacterium bovis e da Brucella abortus durante a cura de queijo tipo parmesão experimentalmente contaminado

Brazilian legislation allows the manufacture of raw milk cheese with a maturation exceeding 60 days at room temperature above 5C, but there is a lack of solid scientific evidence on the efficacy of this maturation process in inactivating important pathogens that may be present in milk, such as Mycobacterium bovis and Brucella abortus. Thus, the objectives of this study were to produce parmesan-type cheese experimentally contaminated with M. bovis and B. abortus and evaluate the survival of these pathogens along 2-month maturation. Parmesan-type cheese was manufactured in the laboratory using whole pasteurized milk with or without inoculation with M. bovis (SB1033) or B. abortus (1119-3) and matured at 18C for up to 63 days. M. bovis was inoculated in Stonebrink-Leslie medium supplemented with antibiotics and incubated at 37C for 45 days, and B. abortus was incubated in Farrel medium at 36C for 3 days. The average D18C value, weighted by variance, was 37.5 ± 5.3 days for M. bovis and 5.9 ± 0.7 days for B. abortus. The average physicochemical parameters in the cheese at the end of the study were as follows: pH = 4.89, water activity = 0.976, and moisture percentage = 43.1%. The pH might have contributed to the reduction in the population of B. abortus but seems not to have influenced the population of M. bovis. We conclude that the duration of the maturation process influences the size of the surviving populations of M. bovis and B. abortus, and that the shortening of the maturation duration might not ensure a decline in pathogen levels to safe levels. Thus, complementary studies considering the effect of several other technological aspects on the survival of these pathogens are required, including the effect of the lactic acid bacterial population, salt content, and temperature of maturation.

A legislação brasileira permite a fabricação queijos de leite cru com maturação superior a 60 dias em temperatura acima de 5C, mas falta evidência científica sólida sobre a eficácia do processo de maturação na inativação de importantes patógenos que podem estar presentes no leite, como o Mycobacterium bovis e a Brucella abortus. Assim, os objetivos desse estudo foram produzir o queijo tipo parmesão experimentalmente contaminado com M. bovis e B. abortus e avaliar a sobrevivência desses patógenos ao longo de 2 meses de maturação. Queijo do tipo parmesão foi fabricado em laboratório com leite pasteurizado integral com ou sem inoculo de M. bovis (SB1033) ou B. abortus (1119-3) e maturado a 18C por até 63 dias. M. bovis foi semeado em meio Stonebrink-Leslie, acrescido de antibióticos, e incubado a 37C por 45 dias e a B. abortus foi semeada em meio Farrel a 36C por 3 dias. O valor D18C médio, ponderado pela variância, foi de 37,5 ± 5,3 dias para o M. bovis e de 5,9 ± 0,7 dias para a B. abortus. Os parâmetros físico-químicos médios do queijo no final do estudo foram como se segue: pH = 4,89, atividade de água = 0,976 e umidade percentual = 43,1%. O pH pode ter contribuído para redução da população de B. abortus mas parece não ter influenciado a população de M. bovis. Concluiu-se que a duração do processo de maturação influencia o tamanho da população sobrevivente de M. bovis e de B. abortus e que o encurtamento do período de maturação pode não garantir um declínio desses patógenos para níveis seguros. Assim, estudos complementares que considerem o efeito de vários outros aspectos tecnológicos na sobrevivência desses patógenos são necessários, incluindo o efeito da população de bactérias ácido láticas, teor de sal e temperatura de maturação.

Antimycobacterial and cytotoxicity activity of microcystins

Background The present work aimed to evaluate the antimycobacterial activity and cytotoxicity of Microcystis aeruginosa toxins, the MC-LR variant and purified extract of [D-Leu1] microcystin-LR. Methods The antimicrobial activity of M. aeruginosa extract and microcystin was evaluated by resazurin microtiter assay against Mycobacterium tuberculosis, M. terrae, M. chelonae and M. kansasii. The cytotoxicity assay was performed by trypan blue exclusion against the HTC cell line. Results Antimicrobial activity was observed in the hexanic extract of M. aeruginosa (RST 9501 strain) against M. tuberculosis, including sensitive and resistant strains with minimal inhibitory concentrations (MIC) between 1.93 M and 0.06 M. The high activity of M. aeruginosa hexanic extract could be attributed to the major presence of the toxins MC-LR and [D-Leu1] MC-LR that showed activity at MIC between 53 and 0.42 M against tested mycobacterial strains. Even at the highest concentration tested, no toxicity of M. aeruginosa extracts was identified against HTC cells. Conclusions These preliminary results suggest that [D-Leu1] MC-LR is a promising candidate for the development of a new antimycobacterial agent.(AU)

Lepromatous leprosy and perianal tuberculosis: a case report and literature review

Leprosy is a chronic infectious disease caused by Mycobacterium leprae, a microorganism that usually affects skin and nerves. Although it is usually well-controlled by multidrug therapy (MDT), the disease may be aggravated by acute inflammatory reaction episodes that cause permanent tissue damage particularly to peripheral nerves. Tuberculosis is predominantly a disease of the lungs; however, it may spread to other organs and cause an extrapulmonary infection. Both mycobacterial infections are endemic in developing countries including Brazil, and cases of coinfection have been reported in the last decade. Nevertheless, simultaneous occurrence of perianal cutaneous tuberculosis and erythema nodosum leprosum is very rare, even in countries where both mycobacterial infections are endemic.(AU)

Lepromatous leprosy and perianal tuberculosis: a case report and literature review

Leprosy is a chronic infectious disease caused by Mycobacterium leprae, a microorganism that usually affects skin and nerves. Although it is usually well-controlled by multidrug therapy (MDT), the disease may be aggravated by acute inflammatory reaction episodes that cause permanent tissue damage particularly to peripheral nerves. Tuberculosis is predominantly a disease of the lungs; however, it may spread to other organs and cause an extrapulmonary infection. Both mycobacterial infections are endemic in developing countries including Brazil, and cases of coinfection have been reported in the last decade. Nevertheless, simultaneous occurrence of perianal cutaneous tuberculosis and erythema nodosum leprosum is very rare, even in countries where both mycobacterial infections are endemic.(AU)

Lepromatous leprosy and perianal tuberculosis: a case report and literature review

Leprosy is a chronic infectious disease caused by Mycobacterium leprae, a microorganism that usually affects skin and nerves. Although it is usually well-controlled by multidrug therapy (MDT), the disease may be aggravated by acute inflammatory reaction episodes that cause permanent tissue damage particularly to peripheral nerves. Tuberculosis is predominantly a disease of the lungs; however, it may spread to other organs and cause an extrapulmonary infection. Both mycobacterial infections are endemic in developing countries including Brazil, and cases of coinfection have been reported in the last decade. Nevertheless, simultaneous occurrence of perianal cutaneous tuberculosis and erythema nodosum leprosum is very rare, even in countries where both mycobacterial infections are endemic.

Occurrence of Mycobacterium bovis and non-tuberculous mycobacteria (NTM) in raw and pasteurized milk in the northwestern region of Paraná, Brazil

Milk is widely consumed in Brazil and can be the vehicle of agent transmission. In this study, was evaluated the occurrence of Mycobacterium bovis and non-tuberculous mycobacteria (NTM) in raw and pasteurized milk consumed in the northwestern region of Paraná, Brazil. Fifty-two milk samples (20 pasteurized and 32 raw) from dairy farms near the municipality of Maringa, Parana State, Brazil were collected. Milk samples were decontaminated using 5% oxalic acid method and cultured on Lowenstein-Jensen and Stonebrink media at 35 °C and 30 °C, with and without 5-10% CO2. Mycobacteria isolates were identified by morphological features, PCR-Restriction Fragment Length Polymorphism Analysis (PCR-PRA) and Mycolic acids analysis. Thirteen (25%) raw and 2 (4%) pasteurized milk samples were positive for acid fast bacilli growth. Nine different species of NTM were isolated (M. nonchromogenicum, M. peregrinum, M. smegmatis, M. neoaurum, M. fortuitum, M. chelonae, M. flavescens, M. kansasii and M. scrofulaceum). M. bovis was not detected. Raw and pasteurized milk may be considered one source for NTM human infection. The paper reinforces the need for intensification of measures in order to avoid the milk contamination and consequently prevent diseases in the south of Brazil.