PCR-based identification of Mycobacterium murphy causing Canine Leproid Granuloma Syndrome in Niterói, southeast Brazil - case report / Identificação baseada em PCR de Mycobacterium murphy causando Síndrome do Granuloma Lepróide Canino em Niterói, sudeste do Brasil ˗ Relato de Caso

Canine Leproid Granuloma Syndrome (CLGS), also known as canine leprosy, is a cutaneous nodular infectious disease caused by Mycobacterium sp.. Despite being reported worldwide, it is still quite unknown and underdiagnosed. Diagnosis may be achieved by cytopathology or histopathology of skin lesions, but identification of the infectious agent is complex, since bacterial in vitro growth is not possible, relying upon molecular techniques such as PCR to confirm Mycobacterium DNA in the sample. We report a CLGS case in Niteroi, Rio de Janeiro state, Brazil, diagnosed by cytopathology and submitted to molecular identification of the agent. PCR amplification of hsp65 gene was performed and revealed 100% genetic homology to M. murphy strain. This is the first CLGS report with molecular identification in Rio de Janeiro state, and this finding should raise awareness about CLGS as a differential diagnosis among granulomatous skin diseases in this region.(AU)

A síndrome de granuloma leproide canino (SGLC), também conhecida como lepra canina, é uma doença infecciosa cutânea nodular causada por Mycobacterium sp. Apesar de ser relatada mundialmente, ainda é bastante desconhecida e subdiagnosticada. O diagnóstico pode ser conseguido por citopatologia ou histopatologia de lesões cutâneas, mas a identificação do agente infeccioso é complexa, uma vez que o crescimento in vitro bacteriano não é possível, dependendo de técnicas moleculares como a PCR para confirmar o DNA de Mycobacterium na amostra. Relatou-se um caso da SGLC em Niterói, estado do Rio de Janeiro, Brasil, diagnosticado por citopatologia e submetido à identificação molecular do agente. Foi realizada amplificação por PCR do gene hsp65, que revelou 100% de homologia genética com a cepa M. murphy. Este é o primeiro relato da SGLC com identificação molecular no estado do Rio de Janeiro, o que mostra a importância de se acrescentar a SGLC ao diagnóstico diferencial das doenças granulomatosas de pele nessa região.(AU)

De la escrófula a la linfoadenitis micobacteriana / From scrophula to mycobacterial lymphadenitis

Resumen En el siglo XIX se pensaba que la tuberculosis y la tumefacción ganglionar cervical llamada escrófula afectaban a individuos predispuestos por una "constitución diatésica" heredada. En 1882 Robert Koch demostró que lesiones tuberculosas y escrofulosas humanas eran causadas por el bacilo Mycobacterium tuberculosis. A principios del siglo XX se estableció que Mycobacterium bovis, bacilo de la tuberculosis del ganado, podía también causar linfoadenitis cervical en humanos, especialmente en niños, por la ingestión de leche de vacas enfermas. La condición disminuyó después que se controló la infección en el ganado y se introdujo la pasteurización de la leche. En 1956 se describió la linfoadenitis cervicofacial granulomatosa necrosante y supurada causada por micobacterias no tuberculosas. Afecta principalmente a niños bajo los cinco años, especialmente en países sin endemia de tuberculosis. Las linfoadenitis cervicales tuberculosas predominan en adultos jóvenes en países con tuberculosis endémica y en individuos infectados por VIH.

In the 19th century it was widely believed that both tuberculosis and cervical lymph node swelling, known as scrophula, affected individuals predisposed to an inherited "diathetic constitution". In 1882 Robert Koch proved that human tuberculosis and scrophulous lesions were caused by the bacillus Mycobacterium tuberculosis. In the early twentieth century it was stated that Mycobacterium bovis, the bacillus of cattle tuberculosis, could also cause cervical lymphoadenitis in humans, especially in children, by the intake of milk from sick cows. The incidence of this condition decreased after the infection was controlled in cattle and pasteurization of the milk was introduced. A type of granulomatous necrotizing and suppurative cervico-facial lymphadenitis associated to non-tuberculous mycobacteria was described in 1956. It mainly affects children younger than 5 years old, particularly those born in countries with non-endemic tuberculosis. Tuberculous cervical lymphadenitis is prevalent in young adults from tuberculosis-endemic countries and in HIV-infected subjects. Infectious etiology displaced the importance of a personal disposition in the development of scrophula. Nevertheless, mutations that confer susceptibility to mycobacterial infection are currently investigated.

Infecção sistêmica por Mycobacterium avium em cão: relato de caso / Systemic Mycobacterium avium infection in a dog: case report

Infecções sistêmicas causadas pelo complexo Mycobacterium avium em cães são consideradas raras. Em cães e gatos, a infecção resulta da ingestão de carne ou do contato com solo ou fômites contaminados. As manifestações clínicas de cães infectados por M. avium tendem a ser vagas ou ausentes, logo o diagnóstico in vivo torna-se difícil. A suspeita de infecção sistêmica por micobacteriose ocorreu, neste relato, após a identificação de bacilos álcool-ácido resistentes na amostra de medula óssea, os quais foram identificados como Mycobacterium avium pelo método molecular de reação em cadeia da polimerase com análise de restrição (PCR-PRA). Este animal apresentava uma aplasia de medula óssea em decorrência de Erhlichia canis, corroborando a maioria dos relatos na literatura em que se associa essa infecção a pacientes imunossuprimidos.

Systemic infections caused by Mycobacterium avium complex are considered rare in dogs. In dogs and cats, the infection comes from eating meat or being in contact with contaminated soil or fomites. Clinical manifestations of dogs infected with M. avium tend to be vague or absent, so the diagnosis "in vivo" becomes difficult. Systemic mycobacterial infection was suspected in this report, after the identification of acid-alcohol resistant bacilli in a bone marrow sample which was identified as Mycobacterium avium by the molecular method Polymerase Chain Reaction - PCR Restriction Analysis (PCR-PRA). This animal had a bone marrow aplasia due to Erhlichia canis corroborating with most reports in the literature that associate this infection with immunosuppressed patients.

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.

First case report of chronic pulmonary lung disease caused by Mycobacterium abscessus in two immunocompetent patients in Saudi Arabia

Mycobacterium abscessus is a rapidly growing non-tuberculous mycobacterium that is a common water contaminant, and an exclusive opportunistic pathogen in immunosuppressed patients; however, it occasionally causes lung infections in immunocompetent persons as well. We report two cases of chronic pulmonary infection caused by M abscessus in two young migrant workers; they were nonreactive to HIV antigen, without any history of pulmonary disorders and no evidence of any immunocompromized status. Both were initially suspected as having pulmonary tuberculosis, and antituberculosis drugs were administered. The preliminary culture found the isolate as a rapidly growing nontuberculous mycobacteria and later by advanced molecular genotyping of the isolates revealed it as M abscessus. The patients were treated with multiple drugs including clarithromycin. The symptoms resolved slowly, the smears and culture became negative, and they recovered completely. This is the first case of its type to be reported from Saudi Arabia

Micobactérias: epidemiologia e diagnóstico / Mycobacteria: epidemiology and diagnosis

Estima-se que um terço da população mundial esteja infectada com Mycobacterium tuberculosis, oque resulta em 2 milhões de mortes anualmente. No mundo, são registrados mais de 8 milhões de novos casos de tuberculose (TB) por ano e o Brasil ocupa o décimo nono lugar dentre os 23 países detentores da maior carga de TB. Os fatores determinantes para o controle desta doença incluem a detecção rápida, a terapia adequada e os meios para que sejam evitadas futuras transmissões. O diagnósticoconvencional (baciloscopia e cultura do microrganismo) apresenta limitações quanto ao tempo de execução e à operacionalidade, visto que o resultado pode levar até 60 dias para ser liberado.Portanto, a importância da detecção precoce do bacilo se torna fundamental para o bloqueio da cadeia de transmissão da TB. Já as micobacterioses, doenças causadas pelas micobactérias não tuberculosas, também estão provocando um importante impacto em razão do aumento dos surtos de infecções cirúrgicas. A identificação rápida e específica destes microrganismos é importante para o diagnóstico e a consequente escolha do tipo de tratamento do paciente, que está diretamente relacionada com a espécie. Portanto, o conhecimento dos agentes etiológicos das doenças causadaspelas micobactérias e o diagnóstico sensível e específico permitem o tratamento adequado e,consequentemente, o bloqueio da cadeia de transmissão da TB e o controle dos surtos relacionados às micobactérias não tuberculosas.

It is estimated that one third of the world population is infected with Mycobacterium tuberculosis, resulting in 2 million deaths annually. More than 8 million new cases of tuberculosis (TB) are registered per year worldwide, and Brazil ranks 19th among the top 23 countries with the highest rates of TB. The determining factors for the control of this disease include early detection, appropriate therapy and measuresfor avoiding transmission. The conventional diagnoses (smear and microorganism culture) have time limitations for implementation and operation, since the result may take up to 60 days to be released. Therefore, early detection is critical for blockingthe chain of TB transmission. Mycobacterial diseases caused by nontuberculous mycobacteria are also having a major impact due to increased outbreaks of surgical infections. Thereby, the rapid and specific identification of microorganisms is important for the diagnosis, which will determine the type of treatment (treatment according to species). Knowledge of etiologic agents of mycobacterial diseases, as well as sensitive and specific diagnosis allows proper treatment by blocking the chain of TB transmission and controlling nontuberculous mycobacteria outbreaks.

Hepatite granulomatosa em bovino causada por Mycobacterium avium subsp. paratuberculosis / Granulomatous hepatitis in cattle caused by Mycobacterium avium subsp. paratuberculosis

Samples from intestines, liver, and lymph nodes were collected from a dairy steer with clinical suspicion of paratuberculosis. The samples were processed for histologic examination with hematoxylin-eosin and Zihel-Neelsen (ZN) staining for the detection of acid-fast bacilli (AFB), and submitted to immunohistochemistry (IHC). Macroscopic changes were observed in the small intestines, with thickening and corrugation of the mucosa. The main microscopic changes were found in small intestines, lymph vessels in the mesentery, and mesenteric lymph nodes characterized by enteritis, lymphangiectasia, and lymphadenitis. Liver presented with granulomatous hepatitis, an uncommon histopathological feature for paratuberculosis. The clinical features associated with positive culture of Mycobacterium avium subsp. paratuberculosis and detection of AFB by ZN and IHC in the cytoplasm of macrophages (epithelioid) in the intestinal mucosa and submucosa, lymph nodes, and liver were important to confirm the diagnosis of paratuberculosis.

Correlación entre PCR en exudado nasal y la reacción de tuberculina para la detección de organismos del complejo Mycobacterium tuberculosis en bovinos / Correlation between PCR and tuberculin test for detection of Mycobacterium tuberculosis complex organisms in bovine cattle

En México, la tuberculosis (TB) es motivo de regionalización del país de acuerdo a la prevalencia, por tal motivo, la comercialización y/o exportación de bovinos, así como su movilización, es restringida. El diagnóstico oficial de la TB en bovinos en campo se realiza con la prueba de tuberculina, y posmorten con histopatología y/o por aislamiento del agente etiológico. Aunque la prueba de la tuberculina tiene baja especificidad, es suficiente para enviar animales reactores a sacrificio, lo que no garantiza que el animal esté infectado. Lo anterior, hace patente la necesidad de métodos más sensibles y específicos. En la presente investigación se analizaron muestras de exudado nasal de bovinos, para detectar microorganismos del complejo Mycobacterium tuberculosis mediante la prueba de reacción en cadena de la polimerasa (PCR). Todos los animales positivos a la tuberculina se consideraron como animales infectados y éstos fueron comparados con los positivos a PCR. Se muestrearon 420 animales provenientes de zonas de alta y baja prevalencia. El ADN de los exudados nasales se obtuvo con el método de extracción bromuro de cetiltrimetilamonio, y éste se utilizó en una PCR anidada para amplificar dos fragmentos (583 pb y 200 pb) de la secuencia IS6110 del complejo M. tuberculosis. La prueba de PCR logró detectar un mayor número de muestras positivas en las zonas de alta prevalencia (57 por ciento), comparada con las zonas de baja prevalencia (20 por ciento). La sensibilidad de la prueba de PCR fue del 90,5 por ciento y la especificidad del 58,3 por ciento, comparada con la prueba de tuberculina. Quedó demostrado que la prueba de PCR en secreciones nasales puede utilizarse en situaciones de compra-venta o en exposiciones ganaderas como un análisis de tamizaje para prevenir el contagio y la introducción de la enfermedad a rebaños sanos.

In Mexico, the bovine tuberculosis (TB) is associated with both regionalization of the country, considering the levels of prevalence of the disease and restrictions in commercialization and/or exportation of cattle. In Mexico, the diagnosis of the bovine tuberculosis is conducted by using the tuberculin test in live animals, bacteriologic isolation and histopathological analysis in postmortem. Although the low sensibility of tuberculin test, is used to sent the animal reactors to slaughter and maybe the animal is not infected. Considering that some diagnosis methods are not sufficiently reliable and specific to detect the presence of the casual agent of this disease, the use of the polymerase chain reaction (PCR) test was evaluated from bovine nasal mucus. For the detection of the Mycobacterium tuberculosis complex, PCR of nasal mucus was better than Caudal-fold Tuberculin Test (CTT). The sensibility of PCR compared with CTT was 90.5% and specifity was 58.3%. The PCR of nasal mucus can use in buy and sell operations and cattle expositions to prevent the contagious and the introduction the pathogens in health cattle.

Enfermedad de Hansen: a propósito de un caso / Hansen`s disease: a purpose of a case

La lepra o enfermedad de Hansen es una enfermedad infecto contagiosa producida por el bacilo Mycobacterium leprae. Presenta dos tipos principales: la lepra tuberculoide, que produce grandes manchas hiperestésicas posteriormente anestésicas, y la lepra lepromatosa, que origina grandes nódulos en la piel. El tratamiento, hoy en día, varía entre seis meses y dos años, y se basa en la administración de sulfonas. Presentamos caso de masculino de 25 años de edad ingresado en el servicio de Medicina Interna del Hospital Sor Juana Inés de la Cruz con el diagnóstico de lepra lepromatosa confirmado posterior a reporte de biopsia de piel.

Resistencia inicial a drogas antituberculosas: Chile, 1997 / Initial resistance to antituberculous drugs in Chile, 1997

Con el objetivo de actualizar la prevalencia de la resistencia inicial (RI) a drogas antituberculosas en Chile, en 1997 se realizó la última determinación nacional de acuerdo a un protocolo estándar recomendado por la Organización Mundial de la Salud (OMS). Se programó estudiar el 50 por ciento de la incidencia de ese año de casos nunca tratados, de tuberculosis pulmonar con baciloscopía positiva. Las pruebas de sensibilidad se realizaron en el Laboratorio de Referencia Nacional de Micobacterias del Instituto de Salud Pública, por el método de las proporciones. Se recibieron 908 cepas, que correspondieron a igual número de enfermos, provenientes de los 26 Servicios de Salud (SS) del país. Luego de excluir 27 cepas por diversas causas, quedaron 881 casos útiles, 732 (83,1 por ciento) nuevos y 149 (16,9 por ciento) con tratamiento previo. La prevalencia global de RI fue de 9,3 por ciento cifra inferior a la de las últimas cuatro determinaciones; su distribución por drogas y asociaciones resultó semejante a la de los estudios previos; la multirresistencia (MR) se encontró en 0,4 por ciento de los casos. La inclusión de 19 pacientes con serología positiva al VIH, no modificó significativamente las cifras. En los enfermos antes tratados se encontró un porcentaje de 18,1 por ciento de resistencia adquirida, semejante a la determinada para ese mismo año en la totalidad de las recaídas y abandonos. La muestra estudiada para RI resultó numérica y cualitativamente representativa de la situación nacional, lográndose así el objetivo de mantener la vigilancia actualizada de su prevalencia en todo el país

Mycobacterium chelonae y Mycobacterium abscessus: patógenos emergentes / Mycobacterium chelonae y Mycobacterium abscessus: emergents pathogens

Mycobacterium chelonae es el nombre correcto de la micobacteria aislada en 1903 de los pulmones enfermos de una tortuga marina. En una especie distinta de Mycobacterium fortuitum, aislado de ranas en 1905, y de Mycobacterium abscessus, considerado actualmente como una subespecie de M. chelonae. Estas tres epecies son las únicas patógenas para el hombre dentro del grupo de micobacterias ambientales o atípicas, de crecimiento rápido, las cuales se caracterizan por formar colonias en cultivo en menos de siete días. Son agetes etiológicos de nódulos y abscesos cutáneos, localizados y diseminados, de lesiones postoperatorias, usualmente en la cicatriz quirúrgica, de lesiones pulmonares y de linfadenitis granulomatosa, de osteomielitis y de queratitis entre otras. Las lesiones cutáneas y de los tejidos blandos son las más frecuentes y resultan generalmente de la inoculación traumática de esta micobacteria. Histológicamente, los nódulos y abscesos muestran un proceso inflamatorio, supurativo y granulomatoso, mixto, en el que en la cuarta parte de los casos pueden demostrarse conglomerados de bacilos ácidos alcohol resistentes, que tienden a estar situados en una vacuola en el centro del absceso. En colombia, se han descrito tres brotes de abscesos subcutáneos producidos por bacterias ambientales, secundarios a la aplicación de inyecciones contaminadas con el germen causal: en 1981, en Bucaramanga, luego de la aplicación de la vacuna contra la fiebre amarilla, en 50 personas, la mayoría niños; en 1989, en Medellín, por la inyección subcutánea de alergenos, en 13 personas; y, en 1993, en varias ciudades de la costa atlántica, luego de aplicaciones subcutáneas de xilocaína, como tratamiento bionergético, en 297 pacientes. Existen otros informes aislados de casos post-traumáticos

Differential identification of pathogenic mycobacteria isolated from patients clinically diagnosed as pulmonary tuberculosis

By staining sputum samples from 121 patients diagnosed clinically to have pulmonary tuberculosis, before and after treating, sputum with digestion decontamination procedure [concentration], acid fast bacilli were detected in 10.7% and 22.36% of the cases respectively with statistically significant difference. By culture on L.J. agar, 38 strains [31.4%] were isolated. Comparing the results of staining, direct smear before and after concentration with that of culture, it was found that Z. N. staining of sputum after concentration is more sensitive and can be relied upon in diagnosing tuberculous infection. Applying the differential identification biochemical reactions, the 38 strains were: 32 [84.2%] Mycobacterium tuberculosis, 2 [5.3%] M. avium, one [2.6%] M. Kansasii, one [2.6%] M. zenopi and 2 strains that could not be identified. All the strains were sensitive to rifampin, 28.9% were resistant to isoniazid, 13.1% were resistant to ethambutol, 7. 9% were resistant to streptomycin and 5.3 were resistant to kanamycin

Mycobacterias potencialmente patogenas de intéres médico

Este estudio de revisión bibliográfico sobre el género mycobacterium, puntualiza avances en la identificación en los aspectos básicos del comportamiento microbiológico, epidemiológico y las características clínico-patológicas principalmente de las especies de mycobacterias potencialmente patógenas de interés médico.

Microbial factors in the pathogenesis of tuberculosis and their potential influence on the protective effect of B.C.G.

This study included 163 tuberculous patients and 43 persons of their healthy contacts. Seventy one strains of mycobacteria were isolated from sputum and only one strain from urine samples. Out of these; 46 strains have been identified as M. tuberculosis and 11 strains as M. bovis based on the results of niacin production, nitrate reduction and TCH susceptibility tests. Ten strains of M. tuberculosis were subjected to the invivo virulence assay of Mitchison. All tested strains were found to be highly virulent. Thus the prevalence of south Indian or low virulent variants of M. tuberculosis in Egypt is not a possibility. The results of tuberculin testing with PPD-B, in the patients and contacts under study, showed a very low prevalence of sensitization to environmental mycobacteria [26. 9% among patients and 27. 9% among contacts]. Accordingly. these findings suggest that the two hypotheses proposed to be among the important causes of failure of protective effect of BCG in the south Indian trial are not present in the Egyptian population sample studied