Genotyping comparison of Mycobacterium leprae isolates by VNTR analysis from nasal samples in a Brazilian endemic region.

This study analyzed the genetic diversity by MIRU-VNTR of Mycobacterium leprae isolates from nasal cavities and related to epidemiological and clinical data. The sample consisted of 48 newly diagnosed leprosy cases that tested positive for M. leprae PCR in nasal secretion (NS) attending to the National Reference Center of Dermatology Dona Libania (CDERM), Fortaleza, Brazil. Total DNA was extracted from NS of each patient and used for amplification of four M. leprae VNTR loci. Four clusters of M. leprae isolates were formed with identical genotypes. In the spatial analysis, 12 leprosy cases presented similar genotypes organized into 4 clusters. The most common genotypes in the current study was AC8b: 8, AC9: 7, AC8a: 8, GTA9: 10, which may represent a genotype of circulating strains most often in Ceará. A minimum set of four MIRU-VNTR loci was demonstrated to study the genetic diversity of M. leprae isolates from NS.

Natural environmental water sources in endemic regions of northeastern Brazil are potential reservoirs of viable Mycobacterium leprae.

BACKGROUND: The detection of live Mycobacterium leprae in soil and animals other than humans suggests that the environment plays a role in the transmission of leprosy. OBJECTIVE: The objective of this study was to investigate the presence of viable M. leprae in natural water sources used by the local population in five municipalities in the state of Ceará, northeastern Brazil. METHODS: Samples were collected from 30 different sources. Viable bacilli were identified by reverse transcriptase polymerase chain reaction (PCR) of the M. leprae gyrA gene and sequencing of the PCR products. Physicochemical properties of each water source were also assessed. FINDINGS: M. leprae gyrA mRNA was found in 23 (76.7%) of the water sources. No association was found between depth of the water and sample positivity, nor was there any association between the type of water used by the population and sample positivity. An association between viable M. leprae and temperature and pH was found. Georeferencing showed a relation between the residences of leprosy cases and water source containing the bacterium. MAIN CONCLUSIONS: The finding of viable M. leprae in natural water sources associated with human contact suggests that the environment plays an important role in maintaining endemic leprosy in the study region.

Natural environmental water sources in endemic regions of northeastern Brazil are potential reservoirs of viable Mycobacterium leprae

BACKGROUND The detection of live Mycobacterium leprae in soil and animals other than humans suggests that the environment plays a role in the transmission of leprosy. OBJECTIVE The objective of this study was to investigate the presence of viable M. leprae in natural water sources used by the local population in five municipalities in the state of Ceará, northeastern Brazil. METHODS Samples were collected from 30 different sources. Viable bacilli were identified by reverse transcriptase polymerase chain reaction (PCR) of the M. leprae gyrA gene and sequencing of the PCR products. Physicochemical properties of each water source were also assessed. FINDINGS M. leprae gyrA mRNA was found in 23 (76.7%) of the water sources. No association was found between depth of the water and sample positivity, nor was there any association between the type of water used by the population and sample positivity. An association between viable M. leprae and temperature and pH was found. Georeferencing showed a relation between the residences of leprosy cases and water source containing the bacterium. MAIN CONCLUSIONS The finding of viable M. leprae in natural water sources associated with human contact suggests that the environment plays an important role in maintaining endemic leprosy in the study region.

Uniform multidrug therapy for leprosy patients in Brazil (U-MDT/CT-BR): Results of an open label, randomized and controlled clinical trial, among multibacillary patients.

BACKGROUND: Leprosy control is based on early diagnosis and multidrug therapy. For treatment purposes, leprosy patients can be classified as paucibacillary (PB) or multibacillary (MB), according to the number of skin lesions. Studies regarding a uniform treatment regimen (U-MDT) for all leprosy patients have been encouraged by the WHO, rendering disease classification unnecessary. METHODOLOGY AND FINDINGS: An independent, randomized, controlled clinical trial conducted from 2007 to 2015 in Brazil, compared main outcomes (frequency of reactions, bacilloscopic index trend, disability progression and relapse rates) among MB patients treated with a uniform regimen/U-MDT (dapsone+rifampicin+clofazimine for six months) versus WHO regular-MDT/R-MDT (dapsone+rifampicin+clofazimine for 12 months). A total of 613 newly diagnosed, untreated MB patients with high bacterial load were included. There was no statistically significant difference in Kaplan-Meyer survival function regarding reaction or disability progression among patients in the U-MDT and R-MDT groups, with more than 25% disability progression in both groups. The full mixed effects model adjusted for the bacilloscopic index average trend in time showed no statistically significant difference for the regression coefficient in both groups and for interaction variables that included treatment group. During active follow up, four patients in U-MDT group relapsed representing a relapse rate of 2.6 per 1000 patients per year of active follow up (95% CI [0·81, 6·2] per 1000). During passive follow up three patients relapsed in U-MDT and one in R-MTD. As this period corresponds to passive follow up, sensitivity analysis estimated the relapse rate for the entire follow up period between 2·9- and 4·5 per 1000 people per year. CONCLUSION: Our results on the first randomized and controlled study on U-MDT together with the results from three previous studies performed in China, India and Bangladesh, support the hypothesis that UMDT is an acceptable option to be adopted in endemic countries to treat leprosy patients in the field worldwide. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00669643.

Uniform multidrug therapy for leprosy patients in Brazil (U-MDT/CT-BR): results of an open label, randomized and controlled clinical trial, among multibacillary patients

BACKGROUND: Leprosy control is based on early diagnosis and multidrug therapy. For treatment purposes, leprosy patients can be classified as paucibacillary (PB) or multibacillary (MB), according to the number of skin lesions. Studies regarding a uniform treatment regimen (U-MDT) for all leprosy patients have been encouraged by the WHO, rendering disease classification unnecessary. METHODOLOGY AND FINDINGS: An independent, randomized, controlled clinical trial conducted from 2007 to 2015 in Brazil, compared main outcomes (frequency of reactions, bacilloscopic index trend, disability progression and relapse rates) among MB patients treated with a uniform regimen/U-MDT (dapsone+rifampicin+clofazimine for six months) versus WHO regular-MDT/R-MDT (dapsone+rifampicin+clofazimine for 12 months). A total of 613 newly diagnosed, untreated MB patients with high bacterial load were included. There was no statistically significant difference in Kaplan-Meyer survival function regarding reaction or disability progression among patients in the U-MDT and R-MDT groups, with more than 25% disability progression in both groups. The full mixed effects model adjusted for the bacilloscopic index average trend in time showed no statistically significant difference for the regression coefficient in both groups and for interaction variables that included treatment group. During active follow up, four patients in U-MDT group relapsed representing a relapse rate of 2.6 per 1000 patients per year of active follow up (95% CI [0·81, 6·2] per 1000). During passive follow up three patients relapsed in U-MDT and one in R-MTD. As this period corresponds to passive follow up, sensitivity analysis estimated the relapse rate for the entire follow up period between 2·9- and 4·5 per 1000 people per year. CONCLUSION: Our results on the first randomized and controlled study on U-MDT together with the results from three previous studies performed in China, India and Bangladesh, support the hypothesis that UMDT is an acceptable option to be adopted in endemic countries to treat leprosy patients in the field worldwide.

Cytokine polymorphisms and susceptibility to leprosy / Polimorfismos de citocinas e susceptibilidade à hanseníase

Cytokines play a key role in the regulation of the immune response against infectious diseases. In leprosy, the polymorphisms of pro-inflammatory cytokine genes may contribute to host susceptibility to Mycobacterium leprae. The objective of this study was to investigate the association between cytokine gene polymorphisms and BCG protection against leprosy in Brazilian leprosy cases and controls. DNA samples were obtained from 46 patients with leprosy and 83 healthy controls (not leprosy contacts). All genotyping (TNF alpha, IFN gamma, IL-6, IL-10 and TGF beta) measurements were taken using sequence-specific primers (SSP) - PCR. When compared to the healthy controls, no significant associations were observed between the cytokine gene polymorphisms studied and their susceptibility to leprosy. The most frequent genotypes in this population were TNF alpha ?G? allele and G/G genotype at position -308, ?A? allele of IFN gamma at position +874, G allele in the IL-6 at position -174, G allele (codon 25) and T/C-G/G genotype in TGF beta, and ?A? allele in IL-10 at position -1082. For those individuals that had a BCG scar, the TGF beta and IFN gamma genotype polymorphisms did not show difference among leprosy patients compared to healthy controls. Polymorphisms of the cytokine genes studied were not associated with an increased occurrence of leprosy.

Mycobacterium leprae in six-banded (Euphractus sexcinctus) and nine-banded armadillos (Dasypus novemcinctus) in Northeast Brazil

Human beings are the main reservoir of the causative agent of leprosy, Mycobacterium leprae. In the Americas, nine-banded armadillos (Dasypus novemcinctus) also act as a reservoir for the bacillus. In the state of Ceará (CE), which is located in Northeast Brazil and is an endemic area of leprosy, there are several species of armadillos, including D. novemcinctus and Euphractus sexcinctus (six-banded armadillo). Contact between humans and armadillos occur mainly through hunting, cleaning, preparing, cooking and eating. This study identified M. leprae DNA in the two main species of armadillos found in Northeast Brazil. A total of 29 wild armadillos (27 D. novemcinctus and 2 E. sexcinctus) were captured in different environments of CE countryside. Samples from the ear, nose, liver and spleen from each of these animals were tested by a nested M. leprae-specific repetitive element polymerase chain reaction assay. The samples that tested positive were confirmed by DNA sequencing. M. leprae was detected in 21% (6/29) of the animals, including five D. novemcinctus and one E. sexcinctus. This is the first Brazilian study to identify the presence of a biomarker of M. leprae in wild armadillos (D. novemcinctus and E. sexcinctus) in a leprosy hyperendemic area where there is continuous contact between humans and armadillos.

Mycobacterium leprae in six-banded (Euphractus sexcinctus) and nine-banded armadillos (Dasypus novemcinctus) in Northeast Brazil.

Human beings are the main reservoir of the causative agent of leprosy, Mycobacterium leprae. In the Americas, nine-banded armadillos (Dasypus novemcinctus) also act as a reservoir for the bacillus. In the state of Ceará (CE), which is located in Northeast Brazil and is an endemic area of leprosy, there are several species of armadillos, including D. novemcinctus and Euphractus sexcinctus (six-banded armadillo). Contact between humans and armadillos occur mainly through hunting, cleaning, preparing, cooking and eating. This study identified M. leprae DNA in the two main species of armadillos found in Northeast Brazil. A total of 29 wild armadillos (27 D. novemcinctus and 2 E. sexcinctus) were captured in different environments of CE countryside. Samples from the ear, nose, liver and spleen from each of these animals were tested by a nested M. leprae-specific repetitive element polymerase chain reaction assay. The samples that tested positive were confirmed by DNA sequencing. M. leprae was detected in 21% (6/29) of the animals, including five D. novemcinctus and one E. sexcinctus. This is the first Brazilian study to identify the presence of a biomarker of M. leprae in wild armadillos (D. novemcinctus and E. sexcinctus) in a leprosy hyperendemic area where there is continuous contact between humans and armadillos.

A demenda de um Centro de Referência Nacional para Hanseníase no Nordeste Brasileiro: por que o excesso de pacientes? / The need for a national reference center in Brazil's northeastern region: why the excess of patients?

Até 2000, o controle de hanseníase no Brasil foi verticalizado. Depois desta data, o processo de descentralização desta doença deu ínicio a ações que priorizaram o nível primário de atenção à saúde. Entretanto, a assistência ainda permanece centralizada em algumas unidades, como o Centro Nacional de Referência em Dermatologia Sanitária D. Libânia (CDERM), em Fortaleza, Ceará. Este centro responde por 84% da detecção do município. O objetivo deste estudo foi investigar os fatores associados à demanda excessiva de casos em nível secundário de atenção representado por este centro de referência. Um estudo transversal foi realizado com 600 usuários selecionados aleatoriamente nos ambulatórios do CDERM. Foram coletados dados: sócio-econômico e demográficos, sobre conhecimento da doença e percepção dos serviços. Oitenta e dois por cento dos participantes tinham baixa situação socio-econômica, 90% vieram encaminhados por outras unidades de saúde e 87% tinham a forma multibacilar. Sessenta e nove por cento receberam atendimento prévio em outras Unidades de Saúde, 49% jamais ouviram falar de hanseníase, 24% referiram sentir medo da doença ou terem sofrido discriminação, 39% dos usuários referiram que o atendimento ininterrupto no horário do almoço favorece a permanência no CDERM, 57% e 27%, respectivamente, referiram que a medicação complementar nunca faltou no CDERM e nas Unidades Básicas de Saúde (UBS). Sessenta e um por cento considerou que o compromisso dos profissionais no CDERM foi ótimo contra 14% nas UBS. Os fatores relatados pelos usuários, especialmente logísticos e de qualidade de atendimento e dos profissionais, poderiam explicar a concentração de usuários neste centro de referência.

A demanda de um centro de referência nacional para hanseníase no nordeste brasileiro: por que o excesso de pacientes? / The need for a national reference center in Brazil's northeastern region: why the excess of patients?

Até 2000, o controle da hanseníase no Brasil foi verticalizado. Depois desta data, o processo de descentralização desta doença deu início a ações que priorizaram o nível primário de atenção à saúde. Entretanto, a assistência ainda permane centralizada em algumas unidades, como o Centro Nacional de Referência em Dermatologia Sanitária D. Libânia (CDERM), em Fortaleza, Ceará. Este centro responde por 84 por cento da detecção do município. O objetivo deste estudo foi investigar os fatores associados à demanda excessiva de casos em nível secundário de atenção representado por este centro de referência. Um estudo transversal foi realizado com 600 usuários selecionados aleatoriamente nos ambulatórios do CDERM. Foram coletados dados: sócio econômico e demográficos, sobre conhecimento da doença e percepção dos serviços. Oitenta e dois por cento dos participantes tinham baixa situação sócio econômica, 90 por cento vieram encaminhados por outras unidades de saúde e 87 por cento tinham a forma multibacilar. Sessenta e nove por cento receberam atendimento prévio em outras Unidades de Saúde, 49 por cento jamais ouviram falar de hanseníase, 24 por cento referiram sentir medo da doença ou terem sofrido discriminação, 39 por cento dos usuários referiram que o atendimento ininterrupto no horário do almoço favorece a permanência no CDERM, 57 por cento e 27 por cento, respectivamente, referiram que a medicação complementar nunca faltou no CDERM e nas Unidades Básicas de Saúde (UBS). Sessenta e um por cento considerou que o compromisso dos profissionais no CDERM foi ótimo contra 14 por cento nas UBS. Os fatores relatados pelos usuários, especialmente logísticos e de qualidade de atendimento e dos profissionais, poderiam explicar a concentração de usuários neste centro de referência.