ABSTRACT Background: Global publications on Q fever have increased after the 2007 epidemic in the Netherlands. However, the epidemiology of Q fever/coxiellosis in Brazil is still poorly understood. Accordingly, there have been few studies investigating the presence of Coxiella burnetii in dairy products around the world, especially in Brazil, where consumption of fresh cheese made from raw-milk is very high. Objective: This study was a random survey to assess the prevalence of C. burnetii by PCR in traditional Minas artisanal cheese from the Serro microregion, Brazil, which is manufactured from bovine raw-milk. Methods: DNA extracted from 53 cheese samples were analyzed by nested PCR with C. burnetii-specific primers and the products confirmed by DNA sequencing. Results: Out of the 53 cheese samples five (9.43%) were C. burnetii DNA-positive, each coming from one of the respective randomly selected manufacturing agroindustries.Based on our results, it is estimated that 1.62 tons/day of ready-to-eat cheese made from raw-milk from a total of 16.2 tons produced daily in the study region are contaminated with C. burnetii. Conclusion: To our knowledge, this is the first report of highly heat-resistant zoonotic pathogen in raw-milk Brazilian artisanal cheese. This food safety hazard has been completely neglected in ready-to-eat raw-milk Brazilian artisanal cheese and could imply potential threats to consumers, since C. burnetii survives in artisanal cheese submitted to long ripening periods. Thus, this work established random and representative baseline prevalence of C. burnetii in this food product in Brazil. Further epidemiological studies, monitoring trends and setting control targets are warranted. Finally, these results point out the importance of including C. burnetii in animal and public health surveillance programs.

RESUMO Objetivo Verificar a ocorrência de resistência a fármacos antituberculose e a associação com características sociodemográficas e clínicas de pacientes de um hospital referência. Métodos Estudo transversal, com dados de pacientes que tiveram a cultura de micobactérias identificada e o respectivo perfil de sensibilidade aos antimicrobianos definido (junho de 2014 a fevereiro de 2016). Foram realizados a análise estatística descritiva e o teste exato de Fisher, para comparação de proporções. Resultados O estudo envolveu 104 pacientes, e todos tiveram resultados para Mycobacterium tuberculosis . A baciloscopia atingiu alta positividade (93,3%), e 15 pacientes (14,4%) apresentaram linhagens resistentes, sendo 6 (5,6%) multirresistentes. As características sociodemográficas e clínicas não foram associadas à resistência. Conclusão A pesquisa permitiu conhecer melhor o perfil dos pacientes com tuberculose e constitui ferramenta para elaboração de políticas públicas específicas. Os pacientes diagnosticados com tuberculose resistente devem ser submetidos à maior supervisão.

ABSTRACT Objective To determine the occurrence of anti-tuberculosis drug resistance and its association with sociodemographic and clinical characteristics of patients in a referral hospital. Methods This was a cross-sectional study based on data from patients who had mycobacterial culture identified and defined antimicrobials sensitivity profile (June 2014 to February 2016). The descriptive statistical analysis and Fisher’s exact test were used to compare proportions. Results The study included 104 patients who had positive results for Mycobacterium tuberculosis . Bacilloscopy had high positivity (93.3%). A total of 15 patients (14.4%) had resistant strains and six (5.6%) multidrug-resistant. The sociodemographic and clinical characteristics were not related with resistance. Conclusion This study contributed to further the understandings about the tuberculosis patients’ profile, the study also served as a tool for development of specific public policies. Patients diagnosed with resistant tuberculosis must be under greater supervision.

RESUMO Objetivo Comparar o desempenho do método de Ogawa-Kudoh ao de Petroff modificado no diagnóstico da tuberculose pulmonar. Métodos Utilizaram-se 205 amostras de escarro de 166 pacientes com suspeita clínica ou controle de tuberculose pulmonar atendidos em um hospital público terciário, entre os meses de julho de 2014 a julho de 2016. Todas as amostras foram processadas simultaneamente pelos métodos de descontaminação Ogawa-Kudoh e Petroff modificado, seguindo as recomendações do Ministério da Saúde. Na análise estatística, foi empregado o teste de McNemar, para comparação de proporções, e o índice Kappa, para verificar o grau de concordância entre os dados. Resultados Os métodos Ogawa-Kudoh e Petroff modificado mostraram-se eficientes na detecção de micobactérias, não sendo verificadas discordâncias significativas tanto nas comparações de pares de resultados (p=0,549), como na taxa de contaminação das culturas (p=0,065). O grau de concordância das técnicas foi considerado excelente (índice Kappa de 0,877), e o Ogawa-Kudoh, em relação ao Petroff modificado, apresentou 90,4% de sensibilidade, 96,6% de especificidade, 94,3% de valor preditivo positivo e 94,2% de valor preditivo negativo. Conclusão O método de Ogawa-Kudoh revelou-se suficientemente sensível e específico para o diagnóstico da tuberculose pulmonar e, portanto, adequado para a aplicação na rotina laboratorial. Por ser mais simples, de baixo custo e com menores exigências técnicas de biossegurança e capacitação profissional, o Ogawa-Kudoh apresenta-se como alternativa para gestores e profissionais da área promoverem a ampliação da cobertura diagnóstica bacteriológica da tuberculose pulmonar.

ABSTRACT Objective To compare the performance of the Ogawa-Kudoh method with the modified Petroff technique in diagnosis of pulmonary tuberculosis. Methods A total of 205 sputum samples from 166 patients with clinical suspicion or under pulmonary tuberculosis follow-up, seen at a public tertiary care hospital, from July 2014 to July 2016 were used. All samples were simultaneously processed using the Ogawa-Kudoh and modified Petroff decontamination methods, according to the recommendations of the Ministry of Health. In the statistical analysis, the McNemar test and the Kappa index were used, respectively, to compare proportions and verify agreement between data. Results The Ogawa-Kudoh and modified Petroff methods were efficient in mycobacteria detection, with no significant differences in results (p=0.549) and contamination rate of the cultures (p=0.065). The agreement between techniques was considered excellent (Kappa index of 0.877) and Ogawa-Kudoh, as compared to the modified Petroff technique, showed sensitivity of 90.4%, specificity of 96.6%, positive predictive value of 94.3% and negative predictive value of 94.2%. Conclusion The Ogawa-Kudoh technique proved to be sufficiently sensitive and specific for diagnosis of pulmonary tuberculosis, and, therefore, suitable for routine laboratory application. Since it is simple, low-cost and has less technical requirements for biosafety and professional training, Ogawa-Kudoh is an alternative for managers and healthcare professionals to promote the expansion of bacteriological diagnostic coverage of pulmonary tuberculosis.

BACKGROUND The World Health Organization (WHO) has classified human zoonotic tuberculosis (TB) due to Mycobacterium bovis as a neglected issue in the developing world. In a recent cross-sectional study in Brazil, three of 189 TB patients presented with a coinfection of M. bovis and M. tuberculosis and were selected as cases for this study. OBJECTIVE The aim was to evaluate risk factors (RF) for zoonotic TB in an urban area of Brazil in order to guide preventive programmes. METHODS A matched case-control study was carried out nested within a cross-sectional study. For each of the three cases, 14 age- and sex-matched controls (TB due to M. tuberculosis) were selected. FINDINGS Zoonotic potential exposures (ZE) and extrapulmonary TB (EPTB) were independently associated with zoonotic TB in multivariate analyses. CONCLUSIONS ZE by occupation and consumption of raw milk and derivative products that place individuals in direct and indirect contact with animals and their excretions/secretions increase the risk for zoonotic TB in Brazil, especially among those with EPTB. Therefore, measures such as efficient control of bovine TB, distribution of pasteurised milk and its derivative products, and the diagnosis and monitoring of zoonotic TB in humans are essential steps, especially in developing countries where bovine TB is enzootic, and further studies are necessary.

OBJETIVO Analisar casos de tuberculose e o impacto do acompanhamento direto na detecção dos desfechos dos tratamentos.MÉTODOS Estudo de coorte aberto prospectivo com 504 casos notificados ao Sistema de Informação de Agravos de Notificação, em Juiz de Fora, MG, no período de 2008 a 2009. Foram comparadas as incidências de encerramentos dos tratamentos de um grupo de casos de tuberculose detectadas por acompanhamento direto (287) por meio de contatos mensais com os pacientes durante os seus retornos, com as coletadas indiretamente (217) por meio do sistema de registro do município. Para comparar as proporções foi utilizado o teste Qui-quadrado, com nível de significância de 0,05. O risco relativo foi utilizado para avaliar a razão de incidências de cada tipo de encerramento entre os dois grupos.RESULTADOS Dos encerramentos acompanhados, direta e indiretamente, 18,5% e 3,2% o abandonaram e 3,8% e 0,5% tiveram falência de tratamento, respectivamente. As incidências de abandono e falência dos tratamentos foram maiores no grupo com acompanhamento direto (p < 0,05) (RR = 5,72; IC95% 2,65;12,34 e RR = 8,31; IC95% 1,08;63,92), respectivamente.CONCLUSÕES Houve maior incidência de abandonos e de falências na população acompanhada diretamente que, em sua maioria, permaneceu negligenciada pelo Sistema de Informação de Agravos de Notificação. São necessárias medidas efetivas para melhoria do controle da tuberculose e da qualidade dos dados.

OBJECTIVE To analyze the cases of tuberculosis and the impact of direct follow-up on the assessment of treatment outcomes.METHODS This open prospective cohort study evaluated 504 cases of tuberculosis reported in the Sistema de Informação de Agravos de Notificação (SINAN – Notifiable Diseases Information System) in Juiz de Fora, MG, Southeastern Brazil, between 2008 and 2009. The incidence of treatment outcomes was compared between a group of patients diagnosed with tuberculosis and directly followed up by monthly consultations during return visits (287) and a patient group for which the information was indirectly collected (217) through the city’s surveillance system. The Chi-square test was used to compare the percentages, with a significance level of 0.05. The relative risk (RR) was used to evaluate the differences in the incidence rate of each type of treatment outcome between the two groups.RESULTS Of the outcomes directly and indirectly evaluated, 18.5% and 3.2% corresponded to treatment default and 3.8% and 0.5% corresponded to treatment failure, respectively. The incidence of treatment default and failure was higher in the group with direct follow-up (p < 0.05) (RR = 5.72, 95%CI 2.65;12.34, and RR = 8.31, 95%CI 1.08;63.92, respectively).CONCLUSIONS A higher incidence of treatment default and failure was observed in the directly followed up group, and most of these cases were neglected by the disease reporting system. Therefore, effective measures are needed to improve the control of tuberculosis and data quality.

OBJETIVO Analisar casos de tuberculose e o impacto do acompanhamento direto na detecção dos desfechos dos tratamentos.MÉTODOS Estudo de coorte aberto prospectivo com 504 casos notificados ao Sistema de Informação de Agravos de Notificação, em Juiz de Fora, MG, no período de 2008 a 2009. Foram comparadas as incidências de encerramentos dos tratamentos de um grupo de casos de tuberculose detectadas por acompanhamento direto (287) por meio de contatos mensais com os pacientes durante os seus retornos, com as coletadas indiretamente (217) por meio do sistema de registro do município. Para comparar as proporções foi utilizado o teste Qui-quadrado, com nível de significância de 0,05. O risco relativo foi utilizado para avaliar a razão de incidências de cada tipo de encerramento entre os dois grupos.RESULTADOS Dos encerramentos acompanhados, direta e indiretamente, 18,5% e 3,2% o abandonaram e 3,8% e 0,5% tiveram falência de tratamento, respectivamente. As incidências de abandono e falência dos tratamentos foram maiores no grupo com acompanhamento direto (p < 0,05) (RR = 5,72; IC95% 2,65;12,34 e RR = 8,31; IC95% 1,08;63,92), respectivamente.CONCLUSÕES Houve maior incidência de abandonos e de falências na população acompanhada diretamente que, em sua maioria, permaneceu negligenciada pelo Sistema de Informação de Agravos de Notificação. São necessárias medidas efetivas para melhoria do controle da tuberculose e da qualidade dos dados.

OBJECTIVE To analyze the cases of tuberculosis and the impact of direct follow-up on the assessment of treatment outcomes.METHODS This open prospective cohort study evaluated 504 cases of tuberculosis reported in the Sistema de Informação de Agravos de Notificação (SINAN – Notifiable Diseases Information System) in Juiz de Fora, MG, Southeastern Brazil, between 2008 and 2009. The incidence of treatment outcomes was compared between a group of patients diagnosed with tuberculosis and directly followed up by monthly consultations during return visits (287) and a patient group for which the information was indirectly collected (217) through the city’s surveillance system. The Chi-square test was used to compare the percentages, with a significance level of 0.05. The relative risk (RR) was used to evaluate the differences in the incidence rate of each type of treatment outcome between the two groups.RESULTS Of the outcomes directly and indirectly evaluated, 18.5% and 3.2% corresponded to treatment default and 3.8% and 0.5% corresponded to treatment failure, respectively. The incidence of treatment default and failure was higher in the group with direct follow-up (p < 0.05) (RR = 5.72, 95%CI 2.65;12.34, and RR = 8.31, 95%CI 1.08;63.92, respectively).CONCLUSIONS A higher incidence of treatment default and failure was observed in the directly followed up group, and most of these cases were neglected by the disease reporting system. Therefore, effective measures are needed to improve the control of tuberculosis and data quality.

Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method.

In this cross-sectional study, mycobacteria specimens from 189 tuberculosis (TB) patients living in an urban area in Brazil were characterised from 2008-2010 using phenotypic and molecular speciation methods (pncA gene and oxyR pseudogene analysis). Of these samples, 174 isolates simultaneously grew on Löwenstein-Jensen (LJ) and Stonebrink (SB)-containing media and presented phenotypic and molecular profiles of Mycobacterium tuberculosis, whereas 12 had molecular profiles of M. tuberculosis based on the DNA analysis of formalin-fixed paraffin wax-embedded tissue samples (paraffin blocks). One patient produced two sputum isolates, the first of which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, and the second of which only grew on SB media and presented phenotypic profiles of Mycobacterium bovis. One patient provided a bronchial lavage isolate, which simultaneously grew on LJ and SB media and presented phenotypic and molecular profiles of M. tuberculosis, but had molecular profiles of M. bovis from paraffin block DNA analysis, and one sample had molecular profiles of M. tuberculosis and M. bovis identified from two distinct paraffin blocks. Moreover, we found a low prevalence (1.6%) of M. bovis among these isolates, which suggests that local health service procedures likely underestimate its real frequency and that it deserves more attention from public health officials.

A cross-sectional analysis of stored Ziehl-Neelsen (ZN)-stained sputum smear slides (SSS) obtained from two public tuberculosis referral laboratories located in Juiz de Fora, Minas Gerais, was carried out to distinguish Mycobacterium bovis from other members of the Mycobacterium tuberculosis complex (MTC). A two-step approach was used to distinguish M. bovis from other members of MTC: (i) oxyR pseudogene amplification to detect MTC and, subsequently, (ii) allele-specific sequencing based on the polymorphism at position 285 of this gene. The oxyR pseudogene was successfully amplified in 100 of 177 (56.5%) SSS available from 99 individuals. No molecular profile of M. bovis was found. Multivariate analysis indicated that acid-fast bacilli (AFB) results and the source laboratory were associated (p < 0.05) with oxyR pseudogene amplification. SSS that were AFB++ SSS showed more oxyR pseudogene amplification than those with AFB0, possibly due to the amount of DNA. One of the two source laboratories presented a greater chance of oxyR pseudogene amplification, suggesting that differences in sputum conservation between laboratories could have influenced the preservation of DNA. This study provides evidence that stored ZN-SSS can be used for the molecular detection of MTC.

O presente trabalho descreve inicialmente um cão com sintomas característicos de leishmaniose visceral. Amostra de soro desse cão foi positiva por imunofluorescência indireta (IFI) conduzida no IgG total anti-Leishmania em 1999. Além disso, tecidos desse cão foram positivos por reação em cadeia pela polimerase (PCR) conduzida em 2004, identificando DNA de Leishmania no cerebelo, fígado, rim e intestino. Esta é a primeira vez que um cão com leishmaniose visceral autóctone foi descrito no Município de Bom Sucesso, Minas Gerais, Brasil. O achado desse cão reagente à IFI levou a uma investigação epidemiológica nesse município. Essa investigação foi conduzida de março de 1999 a dezembro de 2005. Vinte e dois de um total de 734 (3%) cães examinados foram reagentes à IFI. Entre os 22 cães IFI reagentes, seis apresentaram sintomas característicos de leishmaniose visceral canina. Os resultados desta investigação epidemiológica foram enviados às autoridades locais e estaduais de saúde pública requerendo medidas preventivas e de controle para leishmaniose visceral de forma a interromper a transmissão da doença e evitar a ocorrência de casos humanos.

The article begins by describing a dog with characteristic symptoms of visceral leishmaniasis. A serum sample from this animal was positive by indirect immunofluorescence (IIF) performed in anti-Leishmania total IgG in 1999. Tissues from the same dog were also positive by polymerase chain reaction (PCR) in 2004, identifying Leishmania DNA in the cerebellum, liver, kidney, and intestine. This is the first report of a dog with autochthonous visceral leishmaniasis in the county of Bom Sucesso, Minas Gerais State, Brazil. The finding of this IIF-positive dog led to a canine visceral leishmaniasis epidemiological investigation in the county. The investigation was conducted from March 1999 to December 2005. IIF was positive for Leishmania in 22 (3%) of 734 examined dogs. Among the 22 IIF-positive dogs, six presented characteristic symptoms of canine visceral leishmaniasis. The results of this epidemiological investigation were sent to local and State public health authorities, requesting visceral leishmaniasis control and preventive measures to interrupt transmission of the disease and avoid the occurrence of human cases.

O presente trabalho descreve inicialmente um cão com sintomas característicos de leishmaniose visceral. Amostra de soro desse cão foi positiva por imunofluorescência indireta (IFI) conduzida no IgG total anti-Leishmania em 1999. Além disso, tecidos desse cão foram positivos por reação em cadeia pela polimerase (PCR) conduzida em 2004, identificando DNA de Leishmania no cerebelo, fígado, rim e intestino. Esta é a primeira vez que um cão com leishmaniose visceral autóctone foi descrito no Município de Bom Sucesso, Minas Gerais, Brasil. O achado desse cão reagente à IFI levou a uma investigação epidemiológica nesse município. Essa investigação foi conduzida de março de 1999 a dezembro de 2005. Vinte e dois de um total de 734 (3%) cães examinados foram reagentes à IFI. Entre os 22 cães IFI reagentes, seis apresentaram sintomas característicos de leishmaniose visceral canina. Os resultados desta investigação epidemiológica foram enviados às autoridades locais e estaduais de saúde pública requerendo medidas preventivas e de controle para leishmaniose visceral de forma a interromper a transmissão da doença e evitar a ocorrência de casos humanos.

The article begins by describing a dog with characteristic symptoms of visceral leishmaniasis. A serum sample from this animal was positive by indirect immunofluorescence (IIF) performed in anti-Leishmania total IgG in 1999. Tissues from the same dog were also positive by polymerase chain reaction (PCR) in 2004, identifying Leishmania DNA in the cerebellum, liver, kidney, and intestine. This is the first report of a dog with autochthonous visceral leishmaniasis in the county of Bom Sucesso, Minas Gerais State, Brazil. The finding of this IIF-positive dog led to a canine visceral leishmaniasis epidemiological investigation in the county. The investigation was conducted from March 1999 to December 2005. IIF was positive for Leishmania in 22 (3%) of 734 examined dogs. Among the 22 IIF-positive dogs, six presented characteristic symptoms of canine visceral leishmaniasis. The results of this epidemiological investigation were sent to local and State public health authorities, requesting visceral leishmaniasis control and preventive measures to interrupt transmission of the disease and avoid the occurrence of human cases.

Neste estudo, foi comparada a contagem de células somáticas (CCS) de amostras pareadas de leite de 86 cabras pelo método eletrônico (Somacount 300) calibrado com padrão de vaca, com a técnica padrão de contagem microscópica direta utilizando corante pyronina Y - verde de metila. As cabras eram das raças Saanen e Toggenburg de um criatório localizado na Zona da Mata de Minas Gerais. Em complementação, foi avaliado o efeito do conservante Bronopol® no leite de cabras sobre a CCS. Para as 86 amostras de leite sem conservante, a média das leituras de CCS determinadas com o microscópio foi menor e diferiu (p <FONT FACE=Symbol>£</FONT> 0,05) da média de CCS lidas pelo Somacount 300 padronizado com leite de vaca. Porém, a média de leituras de CCS das 86 amostras com Bronopol® lidas no microscópio não apresentou diferença significativa (p>0,05) em relação à obtida com o Somacount 300; e a curva de estimação de CCS no microscópio em função do Somacount foi significativa em nível de 95% com alta correlação, demonstrando a possibilidade de se utilizar o Somacount 300 padronizado com leite de vaca para leituras de CCS de leite de cabras conservado com Bronopol® dentro dos limites de CCS analisados neste estudo.

In this study, there was a comparison of SCC from paired milk samples of 86 goats by the electronic method (Somacount 300) calibrated with cow milk standard, with the pyronin Y-methyl green stain direct microscopic method. The goats were of Saanen and Toggenburg breeds from a farm located at the Zona da Mata of the Minas Gerais State. In addition, was evaluated the effect of Bronopol® on the SCC in goat milk was evaluated. For the unpreserved 86 milk samples, the SCC mean determined with the microscopic method was smaller and differed (p <FONT FACE=Symbol>£</FONT> 0.05) from that obtained with the Somacount 300 calibrated with cow milk. However, the SCC mean of the 86 milk samples with Bronopol® did not differ between the two methods (p>0.05). The estimated SCC curve for the microscopic in function to the Somacount significant with high correlation, demonstrating the possibility of using the Somacount 300 calibrated with cow milk for counting somatic cells in goat milk preserved with Bronopol® within the range of SCC analyzed in this study.