Mycobacterium genavense infection in two species of captive snakes

Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.

Características clínicas, factores de riesgo y perfil de susceptibilidad de las infecciones por micobacterias documentadas por cultivo, en un hospital universitario de alta complejidad en Medellín (Colombia) / Clinical features, risk factors and susceptibility profile of mycobacterial infections documented by culture in a university hospital of high complexity in Medellin (Colombia)

Introduction: Tuberculosis (TB) remains an entity of high prevalence and mortality worldwide. The rising drug resistance is a public health problem. Besides, non-tuberculosis mycobacterial (NTM) infections are described with increasing frequency in areas of high prevalence of TB. Objectives: To determine epidemiological, clinical and microbiological characteristics of mycobacterial infections documented by culture. Materials and Methods: An observational, descriptive study in hospitalized patients. Results: M. tuberculosis complex was identified in 90,9% of 187 patients; 9,1% had NTM, 64% were male and the mean age was 40 years (range 1-88 years). The main co-morbidities were HIV / AIDS (23.5%), use of corticosteroids (13.3%) and chronic kidney disease (9.6%). Clinical forms were pulmonary (56.6%), extra-pulmonary (23.9%) and disseminated (19.2 The most common extra-pulmonary compromise was nodal (7.4%) and gastrointestinal (7%). 10.6% of M. tuberculosis were multi-drugresistant (MDR) and 2.12% had extended drug resistance (XDR). Mycobacterium avium andM. abscessus were the most frequent NTM. Overall mortality was 10%. Conclusions: In our study immune suppression is the main risk factor for extrapulmonary and disseminated disease. Resistance, MDR and XDR is higher in inpatients with TB. MNT infections are not uncommon in our country.

Introducción: Tuberculosis (TBC) es aún una entidad de alta prevalencia y mortalidad en el mundo. La resistencia ascendente a fármacos es un problema de salud pública. Además se describen con mayor frecuencia infecciones por micobacterias no tuberculosas (MNT) en áreas de alta prevalencia de TBC. Objetivos: Determinar características epidemiológicas, clínicas y microbiológicas de las infecciones por micobacterias documentadas por cultivo. Materiales y Métodos: Estudio observacional, descriptivo, en pacientes hospitalizados. Resultados: De 187 pacientes, en 90,9% se identificó complejo M. tuberculosis y en 9,1% MNT; 64% fueron hombres. Edad promedio 40 años (rango 1-88 años). Las principales co-morbilidades fueron infección por VIH/SIDA (23,5%), uso de corticoesteroides (13,3%) y enfermedad renal crónica (9,6%). Las formas clínicas fueron pulmonares (56,6%), extra-pulmonares (23,9%) y diseminadas (19,2%). El compromiso extra-pulmonar más frecuente fue ganglionar (7,4%) y gastrointestinal (7%). En M. tuberculosis 10,6% fueron multidrogoresistentes (MDR) y 2,12% con resistencia extendida (XDR). Mycobacterium avium y M. abscessus fueron las MNT más frecuentes. La mortalidad general fue 10%. Conclusiones: Inmuno-supresión es el principal factor de riesgo para enfermedad extrapulmonar y/o diseminada y la resistencia a fármacos en pacientes hospitalizados con TBC es llamativa, con mayor incidencia de MDR y XDR. Las infecciones por MNT no son infrecuentes en nuestro medio.

DIFFERENTIATION BETWEEN Nocardia spp. AND Mycobacterium spp.: CRITICAL ASPECTS FOR BACTERIOLOGICAL DIAGNOSIS / Diferenciação de Nocardia spp. e Mycobacterium spp.: aspectos críticos para o diagnóstico bacteriológico

New methodologies were developed for the identification of Nocardia but the initial diagnosis still requires a fast and accurate method, mainly due to the similarity to Mycobacterium, both clinical and bacteriologically. Growth on Löwenstein-Jensen (LJ) medium, presence of acid-fast bacilli through Ziehl-Neelsen staining, and colony morphology can be confusing aspects between Nocardia and Mycobacterium. This study describes the occurrence of Nocardia spp. in a mycobacterial-reference laboratory, observing the main difficulties in differentiating Nocardia spp. from Mycobacterium spp., and correlating isolates with nocardiosis cases. Laboratory records for the period between 2008 and 2012 were analyzed, and the isolates identified as Nocardia sp. or as non-acid-fast filamentous bacilli were selected. Epidemiological and bacteriological data were analyzed as well. Thirty-three isolates identified as Nocardia sp. and 22 as non-acid-fast bacilli were selected for this study, and represented 0.12% of isolates during the study period. The presumptive identification was based on macroscopic and microscopic morphology, resistance to lysozyme and restriction profiles using the PRA-hsp65 method. Nocardia spp. can grow on media for mycobacteria isolation (LJ and BBL MGIT™) and microscopy and colony morphology are very similar to some mycobacteria species. Seventeen patients (54.8%) were reported and treated for tuberculosis, but presented signs and symptoms of nocardiosis. It was concluded that the occurrence of Nocardia sp. during the study period was 0.12%. Isolates with characteristics of filamentous bacilli, forming aerial hyphae, with colonies that may be pigmented, rough and without the BstEII digestion pattern in PRA-hsp65 method are suggestive of Nocardia spp. For a mycobacterial routine laboratory, a flow for the presumptive identification of Nocardia is essential, allowing the use of more accurate techniques for the correct identification, proper treatment and better quality of life for patients.

Novas metodologias têm sido desenvolvidas para a identificação de Nocardia spp. mas o diagnóstico inicial ainda necessita de método rápido e preciso, principalmente devido à similaridade com o gênero Mycobacterium, clínica e bacteriologicamente. O crescimento em meio de Löwenstein Jensen (LJ), a presença de bacilos corados pela coloração de Ziehl Neelsen e colônias com características diferentes podem ser fatores de confusão entre nocardias e micobactérias. Este estudo descreve a ocorrência de Nocardia spp. em laboratório de referência em micobacteriologia, observando-se as principais dificuldades em diferenciar Nocardia spp. e Mycobacterium spp., correlacionando isolados com casos de nocardiose. Os registros laboratoriais dos anos 2008 a 2012 foram analisados e os isolados identificados como Nocardia sp. ou como bacilos não álcool - ácido resistentes (NBAAR) foram selecionados. Os dados epidemiológicos e bacteriológicos foram analisados. Trinta e três isolados identificados como Nocardia sp. e 22 como NBAAR foram selecionados para este estudo, perfazendo 0,12% do total de isolados identificados no período estudado. A identificação presuntiva foi baseada na morfologia macroscópica e microscópica, resistência à lisozima e perfis de restrição pelo método PRA-hsp65. Nocardia spp. pode crescer em meios de isolamento para micobactérias (LJ e BBL MGIT™) e microscopia de morfologia e as colônias são muito semelhantes a algumas espécies de micobactérias. Dezessete pacientes (54,8%) foram notificados e tratados para tuberculose, mas apresentaram sinais e sintomas para nocardiose. Concluimos que a ocorrência de Nocardia sp. no período estudado foi de 0,12%. Os isolados com características de bacilos filamentosos, formadores de hifas aéreas, com colônias que podem ter pigmento, rugosas e que não possuem padrão de digestão para BstEII no método PRA-hsp65 são sugestivos de Nocardia spp. Para um laboratório de rotina de Micobactérias, um fluxo de identificação presuntiva para Nocardia spp. é essencial para permitir que esses isolados sejam identificados com técnicas mais precisas, para que seja oferecido o tratamento adequado e qualidade de vida aos pacientes.

Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria

The identification of mycobacteria is essential because tuberculosis (TB) and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA) that was designed for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteria.

Mycobacterium neoaurum causing prosthetic valve endocarditis: a case report and review of the literature

Mycobacterium neoaurum is a rare cause of bacteremia, and infection usually occurs in an immunocompromised host in the setting of an indwelling catheter. Prosthetic valve endocarditis due to non-tuberculous mycobacteria typically carries a dismal prognosis; we report a case ofM. neoaurum prosthetic valve endocarditis with favorable response to antimicrobial therapy without surgical intervention.

Rapid tests for the detection of the Mycobacterium abscessus subsp. bolletii strain responsible for an epidemic of surgical-site infections in Brazil

A single strain of Mycobacterium abscessus subsp. bolletii, characterised by a particular rpoB sequevar and two highly related pulsed field gel electrophoresis patterns has been responsible for a nationwide outbreak of surgical infections in Brazil since 2004. In this study, we developed molecular tests based on polymerase chain reaction restriction-enzyme analysis (PRA) and sequencing for the rapid identification of this strain. Sequences of 15 DNA regions conserved in mycobacteria were retrieved from GenBank or sequenced and analysed in silico. Single nucleotide polymorphisms specific to the epidemic strain and located in enzyme recognition sites were detected in rpoB, the 3' region of the 16S rDNA and gyrB. The three tests that were developed, i.e., PRA-rpoB, PRA-16S and gyrB sequence analysis, showed 100%, 100% and 92.31% sensitivity and 93.06%, 90.28% and 100% specificity, respectively, for the discrimination of the surgical strain from other M. abscessus subsp. bolletii isolates, including 116 isolates from 95 patients, one environmental isolate and two type strains. The results of the three tests were stable, as shown by results obtained for different isolates from the same patient. In conclusion, due to the clinical and epidemiological importance of this strain, these tests could be implemented in reference laboratories for the rapid preliminary diagnosis and epidemiological surveillance of this epidemic strain.

Lung Infection Caused by Mycobacterium riyadhense Confused with Mycobacterium tuberculosis: The First Case in Korea

A slowly growing, non-chromogenic mycobacterial strain was isolated from sputum and bronchial lavage fluid samples of a patient presenting with productive cough, blood-tinged sputum, low-grade fever, and weakness. A positive acid-fast bacilli sputum smear result prompted the initiation of an anti-tuberculosis regimen. Multiplex real-time PCR showed a negative result for Mycobacterium tuberculosis complex and a positive result for nontuberculous mycobacteria. The DNA chip test confirmed this organism as a member of the genus Mycobacterium, but could not specify the species. Interestingly, the mycolic acid patterns obtained by HPLC nearly overlapped with those of M. simulans. The sequences of the Mycobacterium 16S rRNA gene and 16S-23S internal transcribed spacer region were unique and were found to have 100% similarity with those of M. riyadhense. After a review of the literature, we report this case as the first Korean case of M. riyadhense lung infection.

Chronic Pulmonary Disease Due to Mycobacterium monacense Infection: The First Case from Iran

We herein report a case in which the recently characterized species Mycobacterium monacense was isolated from the sputum of an Iranian patient. This case represents the first isolation of M. monacense from Iran. The isolate was identified by conventional and molecular techniques. Our findings show that M. monacense infection is not restricted to developed countries.

Molecular identification and typing of Mycobacterium massiliense isolated from postsurgical infections in Brazil

OBJECTIVE: One hundred thirty-one cases of postsurgical infections were reported in Southern Region of Brazil between August 2007 and January 2008. Thirty-nine (29.8 percent) cases were studied; this report describes epidemiological findings, species identification, antimicrobial susceptibility and clonal diversity of rapidly growing mycobacteria isolated in this outbreak. METHODS: All 39 isolates were analyzed by Ziehl-Nielsen stained smear, bacterial culture and submitted to rpoB partial gene sequencing for identification. The isolates were also evaluated for their susceptibility to amikacin, cefoxitin, clarithromycin, ciprofloxacin, doxycycline, tobramycin and sulfamethoxazole. RESULTS: Thirty-six isolates out of the confirmed cases were identified as Mycobacterium massilienseand the remaining three were identified as Mycobacterium abscessus, Mycobacterium chelonae and Mycobacterium fortuitum. All M. massiliense isolates were susceptible to amikacin (MIC90 = 8 µg/mL) and clarithromycin (MIC90 = 0.25 µg/mL) but resistant to cefoxitin, ciprofloxacin, doxycycline, tobramycin and sulfamethoxazole. Molecular analysis by pulsed-field gel electrophoresis clustered all 36 M. massiliense isolates and showed the same pattern (BRA 100) observed in three other outbreaks previously reported in Brazil. CONCLUSIONS: These findings suggest a common source of infection for all patients and reinforce the hypotheses of spread of M. massiliense BRA100 in Brazilian hospital surgical environment in recent years.

Enfermedad por Mycobacterium simiae y "Mycobacterium sherrisii" en la Argentina / Disease due to Mycobacterium simiae and "Mycobacterium sherrisii" in Argentina

Se presenta información reunida retrospectivamente sobre casos de micobacteriosis originados por Mycobacterium simiae (n = 4) y "M. sherrisii" (n = 6). Los casos ocurrieron entre pacientes con sida (n = 6), historia de silicosis (n = 2) o tuberculosis previa (n = 1). Un caso se perdió luego de diagnosticado y nueve fueron tratados con esquemas terapéuticos basados en claritromicina, etambutol y quinolonas. La respuesta fue muy pobre: cinco pacientes fallecieron (cuatro eran HIV positivos), tres permanecieron crónicos y sólo uno curó. Estas micobacterias originaron 2.1% de los casos de micobacteriosis registrados en un período de ocho años. La distinción de estas micobacterias raras de otras más frecuentes por métodos moleculares rápidos, parece ser clínicamente útil para advertir sobre la dificultad que puede presentar el tratamiento. Sin embargo, la diferenciación genotípica entre M. simiae y "M. sherrisii" parecería no ser clínicamente relevante, dado que no quedaron expuestas características que distingan a los pacientes afectados por los dos microorganismos tan estrechamente relacionados.

A revision of mycobacterial disease due to M simiae (n = 4) and "M. sherrisii" (n = 6) identified during an eight-year period is presented. Cases occurred among patients with AIDS (n = 6), previous history of silicosis (n = 2) or tuberculosis (n = 2). One case was lost to follow-up and the remaining nine responded poorly to chemotherapy based on clarithromycin, ethambutol and fluoroquinolones. Five patients died of whom four were HIV-positive, three remained chronic and one was cured. These microorganisms originated 2.1% of mycobacterioses cases detected in an eight-year period. Timely identification of this group of uncommon mycobacteria by molecular methods seems to be clinically relevant in order to warn of difficulties inherent to the treatment. However, the distinction between both closely related microorganisms might not be crucial for case management as no distinctive characteristics were evident among patients affected by M. simiae or "M. sherrisii".

Infecções por micobactérias de crescimento rápido resistentes a desinfetantes: uma problemática nacional? / Infections by rapidly growing mycobacteria resistant to disinfectants: a national matter?

Rapidly growing mycobacteria (RGM) are opportunistic microorganisms and widely distributed into aqueous environment and soil. Human RGM infections are usually associated with contaminated solutions or medical instruments used during invasive procedures. RGM postsurgical infections have recently emerged in Brazil and have caused national alert, considering the risk factors and epidemiological aspects. This study aimed at analysing the main factors linked to the recent RGM outbreaks, with focus on the national epidemic of Mycobacterium massiliense infections related to the BRA100 strains resistant to 2 percent glutaraldehyde commercial solutions commonly used for preoperative high-level disinfection. Based on previous studies and laboratorial results of assays and colaborations, it has been observed that the cases have been associated with videolaparoscopy for different applications and elective esthetic procedures, such as lipoaspiration and mammary prosthesis implant. Furthermore, outbreaks between 2004 and 2008 and the epidemic in Rio de Janeiro state may be considered particular Brazilian events. Although there are a few epidemiological published studies, some hypotheses based on common aspects related to most national nosocomial occurrences are possible, such as lack of protocols for cleaning and high-level disinfection, use of 2 percent glutaraldehyde as high-level disinfectant for surgical instruments, and dissemination of M. massiliense BRA100 by unknown mechanisms.

Infecção por micobactérias de crescimento rápido após procedimentos videocirúrgicos: a hipótese do glutaraldeído / Rapidly growing mycobacteria infection after videosurgical procedures: the glutaraldehyde hypothesis

Between August 2006 and February 2007, in the state of Rio de Janeiro, Brazil, a massive outbreak of RGM infections after video laparoscopy was mainly associated to the recently described Mycobacterium massiliense species. All confirmed and probable cases reports described the use of high-level disinfection of medical devices by using 2 percent glutaraldehyde (2 percent GA) for 30 min before the surgical procedures. We investigated the susceptibility of the M. massiliense isolates recovered during the outbreak to high-level disinfection after 30 min, 1h, 6h and 10h of exposure to the commercial disinfectants. Reference strains for official mycobactericidal tests such as Mycobacterium abscessus, Mycobacterium bovis, Mycobacterium chelonae, Mycobacterium neoaurum and Mycobacterium smegmatis were included as controls. Although all the reference strains were eliminated in 30 min of exposure to 2 percent GA, we observed the recovery of all M. massiliense clinical isolates even after 10h of exposure. This study suggests that failures in high-level disinfection and the high tolerance of these M. massiliense clinical strains to the 2 percent GA were strongly associated to the magnitude of the outbreak.

Identificación de especies micobacterianas en Cuba / Identification of species mycrobacteriums in Cuba

El aumento de las infecciones producidas por micobacterias ambientales u oportunistas (MAO) coincide en muchos casos con el declive de la infección tuberculosa y el incremento de la infección por el virus de inmunodeficiencia humana (VIH). Sobre todo en los países desarrollados donde se está produciendo un aumento global de la incidencia de enfermedad por MAO, y las micobacteriosis principalmente en pacientes inmunodeficientes cada vez son más frecuentes. En este trabajo se estudiaron 80 cepas recibidas en el Laboratorio Nacional de Referencia e Investigaciones de TB y Micobacterias procedentes de diferentes Centros Provinciales de Higiene y Epidemiología, con el fin de conocer el comportamiento en nuestro país de las especies de interés clínico. Encontramos que al clasificar las micobacterias aisladas según los grupos establecidos por Runyon los siguiente: los grupos con mayor frecuencia fueron el Grupo III y el Grupo IV, por especie las de mayor por ciento de aislamiento fueron: Mycobacterium avium-intracellulare, Mycobacterium fortuitum, Mycobacterium chelonae y Mycobacerium malmoense. Estos estudios son de gran utilidad en los laboratorios de Micobacteriología, pues de esta forma se puede llegar a conocer cuales son las especies predominantes en la población y poder establecer una eficaz vigilancia sobre este tipo de infecciones sobre todo en pacientes inmunodeficientes, grupo más sensibles a estas infecciones.

Micobactérias não-tuberculosas: diversidade das espécies no estado de São Paulo / Nontuberculous mycobacteria: species diversity in São Paulo state, Brazil

O gênero Mycobacterium é constituído por espécies do complexo M. tuberculosis e outras denominadas micobactérias não-tuberculosas (MNT). Até o momento, mais de cem MNT foram descritas. Os objetivos deste estudo foram avaliar a diversidade das espécies de MNT identificadas no estado de São Paulo, no período de 1991 a 1997, que antecedeu a expansão da terapia anti-retroviral, e determinar a freqüência dos casos que atenderam alguns critérios bacteriológicos para o diagnóstico das infecções causadas pelas MNT. MATERIAL E MÉTODOS: Foram analisadas 1.892 cepas isoladas de sítios estéreis e não-estéreis de 1.248 pacientes atendidos no estado de São Paulo. RESULTADOS: Do total de pacientes, 1.199 (96,1 por cento) tiveram suas cepas identificadas e 3,9 por cento apresentaram resultados não-conclusivos. As dez espécies encontradas foram o complexo M. avium (MAC), M. kansasii, M. chelonae, M. fortuitum, M. szulgai, M. xenopi, M. marinum, M. gordonae, M. terrae e M. nonchromogenicum. Quarenta e sete (7,8 por cento) casos pulmonares tiveram diagnóstico confirmado pelo isolamento da mesma espécie em três ou mais amostras e 67 (34 por cento) pacientes tiveram o diagnóstico bacteriológico confirmado por isolamento em sítios estéreis. CONCLUSÕES: As espécies de MNT mais freqüentemente isoladas no estado de São Paulo foram MAC e M. kansasii. Uma publicação nacional com recomendações para diagnóstico e tratamento dessas infecções seria fundamental para a conduta correta no diagnóstico e no tratamento de micobacterioses.

Prevalencia de micobacteriosis y de tuberculosis en pacientes de un hospital de referencia de la provincia de Córdoba / Prevalence of mycobacteriosis and tuberculosis in a reference hospital, Cordoba province

Las micobacterias ambientales (MA) constituyen un importante grupo de especies bacterianas que se encuentran en el medio ambiente, pueden colonizar y ocasionalmente producir enfermedad enel hombre. En este trabajo se investigó la frecuencia de casos de micobacteriosis en relación con los de tuberculosis durante un período de diez años (1.991-2.000). Se estudiaron 16.700 muestras de 9.300 pacientes adultos de ambos sexos asistidos en el Hospital Regional de Tuberculosis de la Provincia de Córdoba, por consulta espontánea. Los aislamientos se realizaron por cultivo en los medios de Lowenstein Jensen y Stonebrink. Las colonias de bacilos ácidoalcohol resistentes (BAAR) se identificaron por pruebas bioquímicas y moleculares. El total de casos diagnosticados fue de 716, de los cuales 684 (95,5%) correspondieron a al complejo Mycobacterium tuberculosis y a micobacterias ambientales 32 (4,5%). Los casos de micobacteriosis se definieron por reiterados aislamientos con desarrollo representativo de una micobacteria ambiental, sospecha clínica y radiológica. De los 32 casos de micobacteriosis, el 75% del total correspondió aMycobacterium avium-intracellulare,15,6% a Mycobacterium fortuitum, 3,1% a Mycobacterium kansasii y 6,3% a Mycobacterium chelonae.Los casos de tuberculosis fueron 94,5% de localización pulmonar y 5,5% extrapulmonar.

Environmental mycobacteria (EM) constitute an important group of bacteria species found in the environment. They can colonize and occasionally produce disease in man. Sixteen thousand three hundred samples from 9300 adult symptomatic patients from the Hospital Regional of Tuberculosis in Cordoba were bacteriolocally investigated. The isolations were performed by culture on Lowenstein Jensen and Stonebrink culture media. The colonies of acid fast bacilli (AFB) were identified by biochemical and molecular tests. Among 716 culture positive cases, 684 (95.5%) were due to Mycobacterium tuberculosis complex and 32 to environmental mycobacteria.Serial samples allowed the confirmation of the etiologicalagent in culture and correlated with consistent clinical and radiological abnormalities. Seventy-five percente of these patients were affected by M. avium complex, 15.6% by M. fortuitum, 3.1% Mycobacterium kansasii and 6.3% Mycobacterium chelonae. Among tuberculosis cases, 94.5% and 5.5% had pulmonary and extrapulmonary disease respectively.

Tracing of pulmonary tuberculosis cases among "goza" smokers using mycobacterial phage typing

The present study included 564 patients with chest symptoms belonging to 3 groups: 175 cigarette smokers, 160 "Goza" smokers [Hubble-bubble or water pipe] and 229 non-smokers. The objective was defining any relation between "Goza" smoking and pulmonary tuberculosis, with special utilization of the technique of Mycobaeteriophage typing in this respect. Results refer to a definite statistically significant increased incidence of pulmonary tuberculosis among "Goza" smokers [19.3%], versus cigarette smokers [5.7%] and non-smokers [4.8%]. There was also statistically significant difference in the clinical and radiological aspects between the 3 groups, namely: art increased incidence of haemoptysis [51.6%] and dyspnea [64.5%] in Goza smokers and more far-advanced extent of the disease among them [70.9%]. The resistance pattern of Mycobacteria isolated from tuberculosis "Goza smokers belonging to the same "Goza-smoking session', was the same, in each of 9 "sessions" from which the cases were collected. The same trend was obtained when the phage typing of the bacilli was correlated with the "Goza-smoking session'. The same phage type, was shown, among the patients collected from the particular session e.g. sessions I .2,3 and 4 showed only phage type A in all their tuberculous patients while session 6 showed phage type B. Phage type Ax was isolated only from session 7 and phage type C only from session 9 and phage type I from session 8. It can he concluded that the mass use of one 'Goza" in the concerned "Goza-smoking session', usually in an unwholesome atmosphere among consumers with low apprehension of symptom like cough and expectoration, is a definite risk factor for cross-infection with pulmonary tuberculosis discovered in an advanced stage. So, "Goza" smokers are a high vulnerable group, in this respect, that needs special epidemiological attention as a public health problem

Application of Infrequent-Restriction-Site Amplification for Genotyping of Mycobacterium tuberculosis and non-tuberculous Mycobacterium

Infrequent restriction site amplification (IRS-PCR) is a method of amplifying DNA sequences, which flank an infrequent restriction site, and produces a strain-specific electrophoretic pattern. We studied the use of IRS-PCR to characterize Mycobacterium tuberculosis and non-tuberculous mycobactria (NTM). One-hundred and sixteen M. tuberculosis and nine NTM isolated at Hanyang University Hospital in Seoul, Korea were used in this study. IRS-PCR using AH1 and PX-G primers produced unique patterns for reference strains, M. tuberculosis H37Rv, M. bovis BCG, M. kansasii, M. scrofulaceum, M. szulgai, M. gordonae, M. avium, M. intracellulae, M. fortuitum, and M. chelonae, respectively. Reference strains M. tuberculosis H37Rv, M. bovis, M. africanum, and all isolates of M. tuberculosis showed similar IRS-PCR patterns. The IRS-PCR patterns generated with multiple isolates of M. tuberculosis from the same patients were essentially identical. IRS-PCR revealed the greatest difference between electrophoretic DNA patterns from M. avium, M. intracellulae, and M. fortuitum that differed from each other and from the reference strains. We concluded that IRS-PCR is a useful tool for strain typing of NTM, but not for M. tuberculosis.