Caracterización clínico-epidemiológica de pacientes con tuberculosis en el Departamento de Caaguazú, Paraguay. 2014 a 2017

Resumen Introducción: La tuberculosis (TBC) es una de las diez principales causas de muerte en todo el mundo. Objetivo: Caracterizar clínica y epidemiológicamente los casos de TBC del Departamento de Caaguazú-Paraguay, entre los años 2014 y 2017. Pacientes y Métodos: Se realizó un estudio observacional, retrospectivo, utilizando datos secundarios del Programa Nacional de Control de la Tuberculosis (PNCT). La población: 659 casos de TBC registrados en el PNCT. Las variables: edad, sexo, grupo poblacional, tipo de TBC, co-infección TBC/VIH, y categoría de egreso. Procesamos la base de datos en Excel 2016 © usando Stata 14.0®. Resultado: El 63,3% de los participantes fue del género masculino. La edad promedio fue de 35,8 años. El 39,6% eran indígenas y 14,5% fueron personas privadas de libertad (PPL) conocidos como reclusos. El 89,6% tuvo TBC pulmonar, 2,4% tuvo co-infección TBC/VIH. La tasa de incidencia fue superior a 21,6/100.000 habts en 2014. La incidencia en indígenas fue de 76,5/100.000 habts en 2017. La incidencia en PPL fue de 2.272,1/100.000 habitantes en 2017. Conclusión: La incidencia de TBC en el Departamento de Caaguazú es baja en la población general afectando principalmente a hombres, mientras que en la población indígena y PPL es alta.

Abstract Background: Tuberculosis (TB) is one of the ten leading causes of death worldwide. Aim: To characterize the clinical and epidemiological point of view of TB cases reported in the Department of Caaguazú-Paraguay, from 2014 to 2017. Methods: Observational, descriptive, retrospective study; Population: 659 cases of TB registered in the National Tuberculosis Control Program (NTCP); variables: age, sex, population group, type of TB, TB/HIV coinfection. We procesed database in Excel 2016 © using Stata 14.0®. Results: 63.3% were of male gender, average age: 35.8 years, 39.6% were indigenous and 85.4% were liberty deprived persons known as inmates (LDP), 89.6% had pulmonary TB and 2,4% had TB/HIV coinfection. Incidence rate exceed 21.6/100,000 inhabitants in 2014. Indigenous incidence was 76.5/100,000 inhabitants in 2017, LDP incidence was 2,272.1/ 100,000 inhabitants in 2017. Conclusion: The incidence of TB in the Department of Caaguazú is low, mainly affecting men, while TB incidence in indigenous people and LDP was high.

Tuberculosis of the central nervous system in cattle in Paraíba, Brazil / Tuberculose no sistema nervoso central de bovinos na Paraíba

This paper describes six cases of tuberculosis in the central nervous system (CNS) of cattle in the state of Paraíba in northeastern Brazil. We reviewed the autopsy reports of 851 bovine necropsies performed from 2003 to 2016. Seventy-three (8.6%) cattle were diagnosed with tuberculosis and six showed lesions in the CNS. Three cases affected cattle up to two-year-old and other three affected adults. Three cattle presented exclusively nervous signs, two had respiratory signs and weight loss and one did not present any clinical signs. At necropsy, five cattle had thickening of the leptomeninges of the cerebellum, pons, obex, spinal cord and cortex, mainly, in the region near the brain basilar Willis´ circle. Another animal, presented a single focal lesion in the cerebellum. Microscopically we observed moderate to severe granulomatous meningitis and encephalitis. Five cattle presented lesions in the lungs and mediastinal lymph nodes and three of them had disseminated lesions in other organs. In all cattle acid-fast bacilli were observed in the lesions and marked positive for immunohistochemistry with polyclonal antibody anti-Mycobacterium tuberculosis. It is concluded that bovine tuberculosis of central nervous system occurs sporadically in Paraíba, in cattle of different ages, most of them with disseminate lesions in other organs. The location of the lesions suggests that the agent invaded the brain by hematogenous route through the circle of Willis.(AU)

Descrevem-se seis casos de tuberculose no sistema nervoso central (SNC) em bovinos, no semiárido da Paraíba. Foram revisados os laudos de um total de 851 necropsias de bovinos realizadas no período de 2003 a 2016. Destes, 73 (8,6%) foram diagnosticados com tuberculose e seis apresentavam lesões no SNC. Três casos ocorreram em bovinos de até dois anos de idade e três em bovinos adultos. Três bovinos apresentaram exclusivamente sinais nervosos, dois tinham sinais respiratórios e perda de peso e um não apresentava nenhum sinal clínico. Macroscopicamente, em cinco bovinos, havia espessamento das leptomeninges do cerebelo, medula espinhal, ponte, obex, colículos e córtex, principalmente, na região basilar encefálica próxima ao polígono de Willis. Em apenas um bovino houve a presença de tubérculo único no cerebelo. Microscopicamente observou-se moderada a acentuada meningite e encefalite granulomatosa. Cinco bovinos apresentaram lesões pulmonares e nos gânglios mediastínicos e três deles tinham lesões disseminadas em outros órgãos. Em todos os bovinos foram encontrados bacilos ácido-álcool resistentes intralesionais e todos tiveram marcação positiva na técnica de imuno-histoquímica com anticorpo policlonal anti-Mycobacterium tuberculosis. Conclui-se que a tuberculose do sistema nervoso central de bovinos ocorre de forma esporádica na Paraíba, principalmente em bovinos com lesões disseminadas em outros órgãos. Sugere-se que a disseminação do agente ocorre pela via hematógena, possivelmente através do polígono de Willis.(AU)

Complete genome sequence of a phthalic acid esters degrading Mycobacterium sp. YC-RL4

ABSTRACT Mycobacterium sp. YC-RL4 is capable of utilizing a broad range of phthalic acid esters (PAEs) as sole source of carbon and energy for growth. The preliminary studies demonstrated its high degrading efficiency and good performance during the bioprocess with environmental samples. Here, we present the complete genome of Mycobacterium sp. YC-RL4, which consists of one circular chromosome (5,801,417 bp) and one plasmid (252,568 bp). The genomic analysis and gene annotation were performed and many potential genes responsible for the biodegradation of PAEs were identified from the genome. These results may advance the investigation of bioremediation of PAEs-contaminated environments by strain YC-RL4.

Granuloma leproide canino: estudo de 27 casos / Canine leproid granuloma: study of 27 cases

O granuloma leproide canino (GLC) é uma doença micobacteriana que cursa com lesão nodular, cutânea ou subcutânea, tipicamente auto limitante, decorrente de infecção pelo Mycobacterium. É uma doença dermatológica rara, usualmente relatada em países de clima tropical. O objetivo deste trabalho é caracterizar as alterações microscópicas e epidemiológicas de casos de GLC. Além disso, utilizar exames complementares para demonstração do Mycobacterium sp. na lesão analisada utilizando coloração de Ziehl-Neelsen, imuno-histoquímica e PCR para classificação e sequenciamento da espécie de Mycobacterium. Foram analisados 27 casos de GLC diagnosticados no período compreendido de 2005 a 2014 (21 exames histológicos e seis citológicos). A raça mais acometida foi Boxer (33,3%), seguida da Dachshund (11,1%). Foi frequente o acometimento de caninos de grande porte (64%) e de pelame curto (96%). Não foi possível estabelecer qualquer padrão de sazonalidade do GLC. As lesões se localizaram anatomicamente no pavilhão auricular na maioria dos casos (93%), constituindo-se de nódulos únicos ou múltiplos, ulcerados ou não. Em nenhum dos casos foi observado acometimento sistêmico. Lesões histológicas seguiram três padrões que variaram de nodulares a difusos, com infiltrado granulomatoso/piogranulomatoso com envolvimento de derme superficial e profunda. A observação de bacilos pela coloração de Ziehl-Neelsen confirmou o diagnóstico. Este teste demonstrou grande variação na quantidade de bacilos nas lesões, não relacionado estatisticamente ao tipo da lesão nem ao período evolutivo. A imuno-histoquímica foi positiva em 90% dos casos, e em alguns casos facilitou a identificação de bacilos, entretanto, em dois casos ocorreram resultados negativos mesmo havendo marcação pela coloração de Ziehl-Neelsen. Na técnica de PCR realizada em 18 amostras, 44% dos casos foram positivos, duas sequências inespecíficas, e demonstrou ainda, que Mycobacterium murphy foi responsável por 2 casos e Mycobacterium hodleri em um caso. Em outros três casos não se obteve 100% de semelhança entre o produto sequenciado e os demais Mycobacterium.(AU)

Canine leproid granuloma (CLG) is cutaneous diseases caused by mycobacteria that course with nodular cutaneous or subcutaneous lesion, typically self-limiting, due to Mycobacterium infection. It is an unusual dermatological disease, usually reported in tropical countries. The objective of the present study is to describe the epidemiological and histopathological findings of CLG. Furthermore, use additional tests for demonstration of Mycobacterium sp. in the lesion were performed, e.g. Ziehl-Neelsen staining, immunohistochemical and polymerase chain reaction (PCR) for classification and genetic sequencing of Mycobacterium species. Twenty seven cases of CLG diagnosed between 2005 and 2014 were studied (21 cases by histological examination and six by cytological examination). Boxer dogs were the breed most affected (33.3%) followed by Dachshund dog breed (11.1%). The disease affected dogs with mainly large size (64%) and short-coated (96%). There was no evidence predilection for occurrence in any season. The lesions were localized, in most cases, at the pinna (93%), comprise one or multiple nodules, ulcerated or not. None of the cases was observed with systemic involvement. Histopathological findings included three patterns of nodular to diffuse, with inflammatory infiltration granulomatous/pyogranulomatous that involved superficial and deep dermis. Definitive diagnosis was made by the presence of acid-fast bacilli by Ziehl-Neelsen stain, showing wide variation in the number of bacilli in the lesions, not being statistically correlated to the type of injury or the evolution period. Immunohistochemistry was effective, and in some cases facilitated the identification of bacilli, but in two cases were negative even with marking by Ziehl-Neelsen staining. The immunohistochemistry was positive in 90% cases, and in some cases facilitated the identification of bacilli, but in two cases were negative even with marking by Ziehl-Neelsen staining. The PCR technique was positive in 44% of cases, demonstrating that Mycobacterium murphy was responsible for two cases of GLC and Mycobacterium hodleri by a one case. In other three PCR-positive cases if not got 100% similarity between the sequenced and other Mycobacterium.(AU)

Infecção por Mycobacterium sp. em herbívoros selvagens de cativeiro no Rio Grande do Sul: estudo retrospectivo e detecção imuno-histoquímica (2003-2015) / Mycobacterium sp. in captive-reared wild herbivores in Rio Grande do Sul: a retrospective study and immunohistochemical detection (2003-2015)

A tuberculose é uma enfermidade infectocontagiosa, debilitante, causada por bacilos álcool-ácido resistentes (BAAR), pertencentes ao complexo Mycobacterium tuberculosis (CMT). As micobacterioses têm importância em Medicina Veterinária devido ao seu potencial zoonótico e sua distribuição mundial, afetando todas as classes de vertebrados. Em animais selvagens as micobacterioses têm sido um problema relatado principalmente em cativeiro. Contudo, há relatos de sua ocorrência também em animais de vida livre, colocando em risco e dificultando os programas de erradicação da tuberculose em animais de produção. O diagnóstico nas espécies selvagens em geral é post mortem, uma vez que o teste de tuberculina não está padronizado para essas espécies, assim como não é confiável para triagem. São consideradas para o diagnóstico lesões de necropsia, observação microscópica de BAAR na coloração de Ziehl-Neelsen (ZN) e, principalmente, isolamento e identificação do agente. No entanto, apenas os achados morfológicos macro e microscópicos não permitem distinguir a espécie de Mycobacterium envolvida. A técnica de imuno-histoquímica (IHQ) com anticorpo policlonal anti-M. tuberculosis confirma a infecção pelo CMT, mas não é específica, pois pode ocorrer marcação de outras micobactérias. As características histológicas, os achados na coloração de ZN e na IHQ de 13 casos de herbívoros selvagens diagnosticados com tuberculose no Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (SPV-UFRGS) no período de 2003 a 2015 são descritos. A partir das amostras em blocos de parafina foram confeccionadas novas lâminas histológicas, coradas com hematoxilina e eosina (HE) e ZN. Cortes foram submetidos à técnica de IHQ para detecção do CMT. Todos os animais eram adultos, provenientes de cativeiro e incluíram lhama (5/13), cervo sambar (4/13), camelo (1/13), cervo vermelho (1/13), anta brasileira (1/13) e antílope Nilgai (1/13). Na IHQ observou-se imunomarcação acentuada (4/13), moderada (4/13) ou discreta (4/13), exceto em um caso, em que não havia quantidade suficiente de material. As características histológicas, bem como os achados na coloração de ZN e na técnica de IHQ confirmaram o diagnóstico de infecção por Mycobacterium sp. e foram considerados métodos rápidos e eficientes, de forma que podem ajudar na prevenção da disseminação da doença em animais.(AU)

Tuberculosis is a debilitating infecto-contagious disease, caused by an acid-fast bacillus (AFB) that belong to different species of the Mycobacterium tuberculosis complex (MTC). Mycobacteriosis are important diseases in veterinary medicine because of their zoonotic potential and worldwide distribution, affecting all classes of vertebrates. In wild animals the mycobacteriosis have been reported mainly as a problem in captivity. There are also reports in free-ranging wildlife, endangering and hampering tuberculosis erradication programs in animal production. The diagnosis of the disease in wildlife is usually postmortem, because the tuberculin test is not standardized for wildlife species, and also it is not reliable for screening. The postmortem diagnosis is based on macroscopic findings, microscopic detection of AFB at Ziehl-Neelsen staining (ZN), and mainly isolation and identification of the agent. However, only gross and microscopic exams do not allow to distinguish the species of Mycobacterium involved. The immunostaining with polyclonal anti-Mycobacterium tuberculosis confirms tuberculosis infection, but is not specific; there may be marking of other mycobacteria. The aim of this study was to describe the histologic findings, of ZN staining and immunohistochemistry technique (IHC) of 13 cases of wildlife herbivores diagnosed with tuberculosis in the Setor de Patologia Veterinária of the Universidade Federal do Rio Grande do Sul (SPV-UFRGS) during the 2003- 2015 period. Formalin fixed paraffin embedded tissues were recut, stained with hematoxylin and eosin and ZN, and samples were submitted to the IHC (anti-M. tuberculosis marking, streptavidin-biotin peroxidase method). All animals included were adults living in captivity and belonged to the following species: llama (5/13), sambar deer (4/13), camel (1/13), red deer (1/13), Brazilian tapir (1/13) and Nilgai antelope (1/13). The IHC revealed immunostaining of accentuated (4/13), moderate (4/13) and discrete (4/13) intensity, except in a case with insufficient material. The histological features, findings in ZN staining and IHC in the wild herbivores with tuberculosis lesions allowed the diagnosis of infection with Mycobacterium sp., turning into fast and efficient methods of diagnosis, which can help to prevent the spread of this disease in animals from the same herd.(AU)

Mutation breeding of high 9α-hydroxy-androst-4-ene-3,17- dione transforming strains from phytosterols and their conversion process optimization / 生物工程学报

In order to improve transformation efficiency of phytosterols into 9α-hydroxylation of 4-androstene-3,17-dione (9α-OH-AD) by Mycobacterium sp. LY-1, we studied the strains breeding using atmospheric and room temperature plasma (ARTP) technology and optimized their conversion process. A high production strain named C33 with a good genetic stability was selected and the product molar yield reached to 15.5%, 34.8% higher than that of original strain with 15 g/L phytosterols. Furthermore, the fermentation medium was optimized through the design of orthogonal experiment. Besides, oil-water bidirectional transformation system was set up to improve the 9α-OH-AD molar yield of mutant strain C33. With adding 12 mL soybean oil to each 1 g phytosterols, the molar yield of 9α-OH-AD reached 47.0%, which increased twice than that of control (15.5%).

Enhancing tuberculosis diagnosis by polymerase chain reaction: an experience at a tertiary hospital

Introduction: Tuberculosis (TB) persists as a severe global public health issue. The aim of the present study was to evaluate the performance of an in-house TB PCR (polymerase chain reaction) in sputum. Methods: DNA from sputum specimens were submitted to a nested-PCR protocol for the IS6110 region detection. PCR results were compared to those of the traditional methods for TB diagnosis, i.e., acid-fast bacilli (AFB) smear microscopy and culture. We analyzed sputum samples obtained from 133 patients. Results: A total of 48 (36%) cultures yielded indeterminate results due to contamination. This high contamination rate may be explained by the fact that samples from fibrocystic patients were included in this study. Additionally, other five samples were positive for nontuberculous mycobacteria (NTM). Therefore, it was possible to compare 80 patients for M. tuberculosis detection. We found 14 positive samples: five presented positive results in the three methods (5/14; 35.7%), two were positive in culture and PCR (2/14; 14.3%), one was positive in AFB and PCR (1/14; 7.1%), five were positive only in PCR (5/14; 35.7%) and 1 was positive only in culture (1/14; 7.1%). Thus, positivity rates for each technique were: 7.5% for AFB (6/80), 10% for culture (8/80) and 16.25% for PCR (13/80). Among the 48 patients who had indeterminate results in sputum culture, two samples were positive in PCR. Conclusion: Considering the limitations of the traditional methods, the use of PCR as a molecular technique could be advantageous for TB diagnosis.

Surgical resection of a cutaneous nodule in the left foot caused by mycobacteria / Exérese cirúrgica de nódulo cutâneo no pé esquerdo causado por uma micobactéria

Nontuberculous mycobacteria are etiologic agents of opportunistic human infections. Although they usually affect superficial tissues, infections in bones and joints have been described. The contamination is associated with increased environmental exposure. With appropriate therapy, the cases usually progress to complete recovery of the patient. This study reports the case of a patient who developed a cutaneous nodule in her left foot acquired when her skin was punctured by a fish. The anatomopathological examination revealed chronic central suppurative granulomatous dermo-hypodermal inflammation. Furthermore, the screening for resistant acid-fast bacilli was positive.

Micobactérias não tuberculosas são agentes causadores de infecções oportunistas no homem. Embora, geralmente, afetem tecidos superficiais, infecções em ossos e articulações têm sido descritas. O contágio está associado ao aumento da exposição do homem ao meio ambiente. Diante da terapêutica adequada, os casos normalmente evoluem com recuperação total do indivíduo. Este estudo descreve o caso clínico de um paciente que apresentou nódulo cutâneo no pé esquerdo após traumatismo com um peixe. O exame anatomopatológico revelou inflamação crônica granulomatosa centralmente supurativa dermo-hipodérmica e a pesquisa de bacilos álcool-ácido resistentes (BAAR) foi positiva.