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ABSTRACT Tuberculosis (TB) and infectious diseases caused by non-tuberculous mycobacteria (NTM) are global concerns. The development of a rapid and accurate diagnostic method, capable of detecting and identifying different mycobacteria species, is crucial. We propose a molecular approach, the BiDz-TB/NTM, based on the use of binary deoxyribozyme (BiDz) sensors for the detection of Mycobacterium tuberculosis (Mtb) and NTM of clinical interest. A panel of DNA samples was used to evaluate Mtb-BiDz, Mycobacterium abscessus/Mycobacterium chelonae-BiDz, Mycobacterium avium-BiDz, Mycobacterium intracellulare/Mycobacterium chimaera-BiDz, and Mycobacterium kansasii-BiDz sensors in terms of specificity, sensitivity, accuracy, and limit of detection. The BiDz sensors were designed to hybridize specifically with the genetic signatures of the target species. To obtain the BiDz sensor targets, amplification of a fragment containing the hypervariable region 2 of the 16S rRNA was performed, under asymmetric PCR conditions using the reverse primer designed based on linear-after-the-exponential principles. The BiDz-TB/NTM was able to correctly identify 99.6% of the samples, with 100% sensitivity and 0.99 accuracy. The individual values of specificity, sensitivity, and accuracy, obtained for each BiDz sensor, satisfied the recommendations for new diagnostic methods, with sensitivity of 100%, specificity and accuracy ranging from 98% to 100% and from 0.98 to 1.0, respectively. The limit of detection of BiDz sensors ranged from 12 genome copies (Mtb-BiDz) to 2,110 genome copies (Mkan-BiDz). The BiDz-TB/NTM platform would be able to generate results rapidly, allowing the implementation of the appropriate therapeutic regimen and, consequently, the reduction of morbidity and mortality of patients.IMPORTANCEThis article describes the development and evaluation of a new molecular platform for accurate, sensitive, and specific detection and identification of Mycobacterium tuberculosis and other mycobacteria of clinical importance. Based on BiDz sensor technology, this assay prototype is amenable to implementation at the point of care. Our data demonstrate the feasibility of combining the species specificity of BiDz sensors with the sensitivity afforded by asymmetric PCR amplification of target sequences. Preclinical validation of this assay on a large panel of clinical samples supports the further development of this diagnostic tool for the molecular detection of pathogenic mycobacteria.

Abstract Infection disease caused by nontuberculous mycobacteria (NTM) have been increasingly reported and often manifest with the same symptoms of tuberculosis (TB). The identification of the causative agent is fundamental for the determination of an appropriate therapy, since each species of mycobacteria requires a specific treatment. Thus, rapid and accurate tests to identify mycobacteria of medical interest, such as Mycobacterium tuberculosis complex (MTBC) and Mycobacterium avium complex (MAC), are necessary in the clinical routine. The present study evaluated an in-house multiplex PCR to detect, in reference and clinical strains, the genus Mycobacterium and also M. tuberculosis and MAC. To identify the Mycobacterium genus, M. tuberculosis and MAC, it was amplified a fragment of hsp65 gene, esat-6 gene, and the internal transcribed spacer between the 16S and 23S rRNA genes, respectively. In total, 87 mycobacteria strains were used, being 10 reference and 77 clinical strains, previously identified as MTBC (n = 66), M. avium (n = 8) or other NTM specie (n = 13). The hsp65 gene fragment was amplified for all mycobacteria strains evaluated (87/87). This multiplex PCR presented sensitivity of 100% and specificity of 95.2% for M. tuberculosis detection, and sensitivity of 100% and specificity of 100% for MAC detection. The multiplex PCR evaluated is an important tool for the differentiation between M. tuberculosis and NTM, as well as for the identification of MAC, a complex composed by species with high prevalence in the world.

Background and objectives: to understand the impact of the COVID-19 pandemic on tuberculosis (TB) diagnosis in different settings is essential to guide the establishment of appropriate TB control strategies. This study aimed to assess the influence of COVID-19 pandemic in laboratory diagnosis of TB in patients tested and diagnosed for TB. Methods: a data survey was carried out in the database of laboratories that perform TB diagnosis for the public health system in Rio Grande city (Rio Grande do Sul, Brazil). Results: there was a decrease of 1,368 to 735 (reduction of 46.3%) in the number of patients tested for TB in public diagnostic services in 2019 and 2020, respectively, and a decrease of 197 to 119 (reduction of 39.6%) in the number of new TB cases diagnosed. In contrast, the positivity rate was 14.4% in 2019 and 16.2% in 2020. Moreover, it was observed that the laboratory that performs the diagnostic service for Primary Health Care was the most affected, when compared to Tertiary Health Care. Conclusion: as a consequence of measures to control the spread of SARS-CoV-2, there was a reduction in TB testing and in the detection of new cases, especially in Primary Health Care, where patients with less need for hospitalization are received.

Background: Due to the difficulty in the access to free-ranging birds, data regarding Aspergillus infections in wild avian species is rare compared to captive wild and domestic birds. Objective: report three cases of Aspergillus section Fumigati causing fungal disease in free-ranging aquatic birds, with the identification of the causal agent to the species level. Case reports: The diagnosis of aspergillosis was performed by macroscopic lesions found during the necropsy and confirmed by culture. Molecular identification by partial sequencing of the calM and benA genes allowed to confirm Aspergillus fumigatus sensu stricto as the etiological agent of aspergillosis in Procellaria aequinoctialis (White-chinned petrel) (n = 1), Nannopterum brasilianus (Neotropical cormorant) (n = 1) and Chroicocephalus maculipennis (Brown-hooded gull) (n = 1). Conclusion: Larger studies regarding the importance of aspergillosis in free-ranging aquatic birds are necessary, as well as it potential role in the One Heath context.

ABSTRACT Human toxocariasis consists of chronic tissue parasitosis that is difficult to treat and control. This study aimed to evaluate the action of the probiotic Lactobacillus acidophilus ATCC 4356 on larvae of Toxocara canis and the effect of IFN-γ cytokine on parasite-host in vivo (1.109 CFU) and in vitro (1.106, 1.107, 1.108, 1.109 CFU) interactions. Four groups of six BALB/c mice were formed: G1 - L. acidophilus supplementation and T. canis infection; G2 - T. canis infection; G3 - L. acidophilus supplementation; and G4 - PBS administration. Mice were intragastrically suplemented with probiotics for 15 days before inoculation and 48 h after inoculation with 100 T. canis eggs. The inoculation of T. canis was also perfomed intragastrically. The recovery of larvae took place through digestion of liver and lung tissues; the evaluation of IFN-γ gene transcription in leukocytes was performed by qPCR. The in vitro test consisted of incubating the probiotic with T. canis larvae. The supplementation of probiotics produced a reduction of 57.7% (p = 0.025) in the intensity of infection of T. canis larvae in mice, whereas in the in vitro test, there was no larvicidal effect. In addition, a decrease in the IFN-γ gene transcription was observed in both, T. canis-infected and uninfected mice, regardless of whether or not they received supplementation. The probiotic L. acidophilus ATCC 4356 reduced T. canis infection intensity in mice, however, the probiotic did not have a direct effect on larvae, demonstrating the need of interaction with the host for the beneficial effect of the probiotic to occur. Yet, the proinflammatory cytokine IFN-γ did not apparently contributed to the observed beneficial effect of probiotics.

Abstract Coronavirus disease (COVID-19) is a pandemic caused by a new coronavirus, called SARS-CoV-2. This disease was first identified in December 2019 and rapidly developed into a challenge to the public health systems around the world. In the absence of a vaccine and specific therapies, disease control and promotion of patient health are strongly dependent on a rapid and accurate diagnosis. This review describes the main laboratory approaches to making a diagnosis of COVID-19 and identifying those previously infected with SARS-CoV-2.

Abstract (1) Background: The Commercial Kit SIRE Nitratase® PlastLabor, is a drug susceptibility test kit used to detect Mycobacterium tuberculosis resistance to first-line TB treatment drugs. The present study aimed at evaluating its performance in a multicenter study. (2) Methods: To determine its accuracy, the proportion methods in Lowenstein Jensen medium or the BACTECTMMGITTM960 system was used as a gold standard. (3) Results: The study revealed that the respective accuracies of the kit with 190 M. tuberculosis clinical isolates, using the proportion methods in Lowenstein Jensen medium or BACTECTMMGITTM960 system as a gold standard, were 93.9% and 94.6%, 96.9% and 94.6%, 98.0% and 97.8%, and 98.0% and 98.9%, for streptomycin, isoniazid, rifampicin, and ethambutol, respectively. (4) Conclusion: Thus, the kit can rapidly screen resistance to streptomycin, isoniazid, rifampicin, and ethambutol. Additionally, it does not require sophisticated equipment; hence, it can be easily used in the laboratories of low and middle income countries.

ABSTRACT Objective: Nontuberculous mycobacteria (NTM) are a heterogeneous group of bacteria that are widely distributed in nature and associated with opportunistic infections in humans. The aims of this study were to identify NTM in patients with suspected tuberculosis who presented positive cultures and to evaluate the genetic diversity of strains identified as Mycobacterium avium. Methods: We studied pulmonary and extrapulmonary samples obtained from 1,248 patients. The samples that tested positive on culture and negative for the M. tuberculosis complex by molecular identification techniques were evaluated by detection of the hsp65 and rpoB genes and sequencing of conserved fragments of these genes. All strains identified as M. avium were genotyped using the eight-locus mycobacterial interspersed repetitive unit-variable-number tandem-repeat method. Results: We found that NTM accounted for 25 (7.5%) of the 332 mycobacteria isolated. Of those 25, 18 (72%) were M. avium, 5 (20%) were M. abscessus, 1 (4%) was M. gastri, and 1 (4%) was M. kansasii. The 18 M. avium strains showed high diversity, only two strains being genetically related. Conclusions: These results highlight the need to consider the investigation of NTM in patients with suspected active tuberculosis who present with positive cultures, as well as to evaluate the genetic diversity of M. avium strains.

Abstract INTRODUCTION: Previous studies that detected Paracoccidioides spp. DNA in soil taken from rural areas have shown this to be a valuable tool for ecological and epidemiological studies. This study reports the detection of Paracoccidioides spp. DNA in soil samples from an urban area of southern Brazil. METHODS: Sixteen soil samples were submitted to nested-PCR and the amplicons of a representative number of positive samples were sequenced. RESULTS: Paracoccidioides spp. DNA was found in 44% of samples. Four DNA amplicons were sequenced, showing 100% homology with P. brasiliensis. CONCLUSIONS: The southern Brazilian urban population is commonly exposed to the Paracoccidioides fungus.

O aumento de casos de esporotricose zoonótica tornou-se um problema de saúde pública em áreas hiperendêmicas para a doença em felinos. O envolvimento dos gatos domésticos nesta transmissão e o contato contínuo destes animais com o solo sugerem que o ambiente possa ser uma importante fonte e/ou reservatório de fungos Sporothrix spp. Assim, este estudo objetivou avaliar a presença de fungos do complexo Sporothrix em amostras de solo provenientes de locais de acesso de felinos infectados no sul do Brasil. Foram coletadas amostras de solo de residências com felinos com esporotricose, de locais próximos a estas residências e também de locais públicos, totalizando 101 amostras. Estas foram processadas pela técnica de plaqueamento direto e incubadas a 25°C por até 15 dias. Dezessete amostras foram paralelamente submetidas a extração de DNA (kitNORGEN BIOTEK CORP(r)) e técnica de PCR espécie-específico e Nested-PCR. Embora em nenhuma amostra avaliada tenha sido detectada a presença de Sporothrix spp., seja por cultivo ou por biologia molecular, este estudo não descarta o papel do ambiente na tríade do processo infeccioso (solo - animal - humano), sendo necessários outros estudos ampliando área de abrangência, volume, tipo e método de coleta das amostras, bem como técnicas de detecção.Palavras-chave: Ambiente; geofílico; S. brasiliensis; S. schenckii.

Helicobacter pylori-induced gastric mucosal inflammation is mediated by proinflammatory and anti-inflammatory cytokines. Polymorphisms in genes that code cytokines influence cytokine secretion levels and appear to contribute to the risk of gastric diseases. In this sense, we performed this study to identify the polymorphisms in the IL-6, IL-8, and IL-10 genes and their associations with H. pylori infection and gastric pathologies. Methods: Gastric biopsy samples of 151 patients infected with H. pylori and 76 uninfected individuals were used. Helicobacter pylori infection was diagnosed by histological examination and the detection of the ureA and glmM genes. The polymorphisms in the IL-6 (at position −174), IL-8 (at position −251), and IL-10 (at position −819) were detected by polymerase chain reaction–restriction fragment length polymorphism. Results: Among the genetic polymorphisms studied, we observed that only the presence of the A allele at position −251 of the IL-8 gene was significantly associated with H. pylori infection. In addition, patient carriers of the A/A genotype at position −251 of the IL-8 gene and carriers of the T allele at position −819 of the IL-10 gene had an increased risk of peptic ulcer disease in the presence of H. pylori infection. We did not find a correlation between polymorphisms in the IL-6, IL-8, and IL-10 genes and a higher risk of gastric carcinoma. Conclusion: We demonstrated that polymorphisms in the IL-8 gene was significantly associated with H. pylori infection. Furthermore, polymorphisms in the IL-8 and IL-10 genes were associated with an enhanced risk of peptic ulcer disease in H. pylori-positive patients.

Background and Objectives: Helicobacter pylori is linked to gastroduodenal pathologies. To determine the frequency and potential risk factors of the H. pylori infection. Material and methods: A cross-sectional study was conducted, including 227 patients, submitted to upper gastrointestinal endoscopy. A questionnaire was applied to the patients, before endoscopy. The biopsy specimens were obtained from the antrum and gastric body for histology and PCR. The chi-square test was used for the categorical data analysis. P-values0.05). However, we observed a significant association between the number of persons per household and presence of H. pylori (p=0.04). A statistically significant relation also was found between H. pylori and the patient’s age (p=0.04) and between the histological and endoscopic diagnoses and the H. pylori infection (p≤0.01). Conclusions: We found a significant relation between household crowding and presence of H. pylori, which seems facilitate the person-to-person transmission H. pylori within families. Our results also suggest a cohort phenomenon. The increase in the frequency of H. pylori infection according to age may be due the acquisition of bacterium predominantly in childhood, when the sanitary conditions were deficient, and not during adulthood. Once acquired and untreated, the persistent H. pylori infection might have led to the development of severe gastroduodenal diseases.

Justificativa e objetivos: As fluoroquinolonas representam uma classe de antimicrobianos que é frequentemente prescrita no tratamento de infecções do trato urinário (ITU), tanto de origem hospitalar como originárias da comunidade. Este estudo tem como objetivo,determinar a frequência de resistência à ciprofloxacina e levofloxacina, além de seus fatores associados em bacilos Gram negativos (BGN) isolados de pacientes com ITU em um hospital no Sul do Brasil. Métodos: foi realizado um estudo transversal e analítico, baseado em casos de infecções do trato urinário causadas por bactérias gram-negativas provenientes de pacientes atendidos no Hospital Universitário Dr. Miguel Riet Correa Jr. em Rio Grande/RS, de agosto de 2012 a julho 2013. Foram analisadas variáveis independentes como idade, sexo, origem da infecção e características dos isolados clínicos bacterianos. Resultados:Dos 562 BGN isolados e analisados, foi encontrado uma frequência de resistência à ciprofloxacina e à levofloxacina de 25,5 e 23,3%, respectivamente, 62,6% de origem comunitária e 59% de origem hospitalar. Os fatores de risco associados à resistência a ciprofloxacina e levofloxacina, foram pacientes do gênero masculino, infecções adquiridas no ambiente hospitalar, commaior tempo de internação, e apresença de β-lactamases de espectro estendido (ESBL) nos isolados clínicos. Conclusões:Os resultados mostraram uma forte associação da resistência bacteriana de BGN com a permanência no ambiente hospitalar e a presença de ESBL. A fim de controlar a resistência antimicrobiana e a redução nos custos em saúde pública torna-se necessário que os hospitais tenham uma forte política de vigilância contínua do uso e da resistência de antibióticos.

Introducción. Los informes sobre Mycobacterium tuberculosis resistente a múltiples fármacos están aumentando a nivel mundial y los laboratorios toman cada vez más conciencia sobre la necesidad de realizar pruebas de sensibilidad a fármacos. Debido al tiempo prolongado que se requiere para hacerlas, se han propuesto nuevos enfoques para la detección rápida de resistencia a los medicamentos, tales como la prueba de la nitrato reductasa. Objetivo. En este estudio se propuso una prueba de sensibilidad a fármacos colorimétrica, rápida y directa, en un formato de microplacas utilizando medio sólido. Materiales y métodos. La precisión del diagnóstico se evaluó mediante la comparación del método de las proporciones con la prueba de la nitrato reductasa directa en placas. Se descontaminaron muestras positivas congeladas de esputo y se procesaron mediante el método de Petroff. La suspensión ya descontaminada se utilizó para hacer la prueba de la nitrato reductasa directa en medio 7H11, utilizando 1 μg/ml de rifampicina, 0,2 μg/ml de isoniacida, 2 μg/ml de ofloxacina, 6 μg/ml de de kanamicina, 2 μg/ ml de amicacina y 10 μg/ml de capreomicina. Se analizaron 84 muestras y los resultados del 69 % de ellas estuvieron disponibles en 21 días. Resultados. La sensibilidad y la especificidad de la prueba comparada con el método de las proporciones fueron de 98,5 y 100 % para la isoniacida, de 98,3 y 96,2 % para la rifampicina, de 91,7 y 100 para la kanamicina, de 78,8 y 97,3 % para la ofloxacina y de 100 y 100 % para la amikacina y la capremicina, respectivamente. Conclusión. Este nuevo formato de la prueba de la nitrato reductasa directa es un método preciso, rápido y económico para determinar el perfil de sensibilidad de M. tuberculosis y puede convertirse en una alternativa para los países con recursos limitados.

Background and objectives: Despite its use as a genotyping method for different microorganisms, few studies have reported the use of Multiple-Locus Variable Number of Tandem Repeats Analysis (MLVA) for the analysis of the clonal diversity of Helicobacter pylori. The aim of this study was to determine the genetic variability of H. pylori strains by MLVA in southern Brazil. Methods: 95 H. pylori DNA samples were obtained from the gastric biopsies of H. pylori-positive patients from two cities in southern Brazil and their clonal analysis was evaluated by MLVA method using electrophoresis in agarose gel. For selection of loci to be analyzed in this study, an in silico analysis of 12 loci previously described in the literature was performed. Results: From the in silico analysis, only four loci were viable for the genotypic analysis of these H. pylori strains, resulting in 90 strains distributed in eight different groups and five orphan strains. Conclusion: Despite MLVA method allow make inferences about the genotypic diversity of a population, our results showed that H. pylori genotyping methods should be critically evaluated prior to their use in this region of Brazil.

Abstract: INTRODUCTION: The World Health Organization endorses the BACTEC Mycobacterial Growth Indicator Tube (MGIT)(tm) system as a rapid, sensitive, and specific method to diagnostic of tuberculosis. Here, we compared the performance of this system against Ogawa-Kudoh cultures and microscopy. METHODS: A total of 927 samples were obtained between December 2011 and December 2013 from 652 cases of suspected tuberculosis at the School Hospital of the Federal University of Rio Grande in Brazil. RESULTS: The MGIT system confirmed tuberculosis in more cases in less time. CONCLUSIONS: The MGIT system is an effective tool for early diagnosis of tuberculosis, especially in patients with HIV/AIDS.

Numerous studies show efflux as a universal bacterial mechanism contributing to antibiotic resistance and also that the activity of the antibiotics subject to efflux can be enhanced by the combined use of efflux inhibitors. Nevertheless, the contribution of efflux to the overall drug resistance levels of clinical isolates of Mycobacterium tuberculosis is poorly understood and still is ignored by many. Here, we evaluated the contribution of drug efflux plus target-gene mutations to the drug resistance levels in clinical isolates of M. tuberculosis. A panel of 17 M. tuberculosis clinical strains were characterized for drug resistance associated mutations and antibiotic profiles in the presence and absence of efflux inhibitors. The correlation between the effect of the efflux inhibitors and the resistance levels was assessed by quantitative drug susceptibility testing. The bacterial growth/survival vs. growth inhibition was analyzed through the comparison between the time of growth in the presence and absence of an inhibitor. For the same mutation conferring antibiotic resistance, different MICs were observed and the different resistance levels found could be reduced by efflux inhibitors. Although susceptibility was not restored, the results demonstrate the existence of a broad-spectrum synergistic interaction between antibiotics and efflux inhibitors. The existence of efflux activity was confirmed by real-time fluorometry. Moreover, the efflux pump genes mmr, mmpL7, Rv1258c, p55, and efpA were shown to be overexpressed in the presence of antibiotics, demonstrating the contribution of these efflux pumps to the overall resistance phenotype of the M. tuberculosis clinical isolates studied, independently of the genotype of the strains. These results showed that the drug resistance levels of multi- and extensively-drug resistant M. tuberculosis clinical strains are a combination between drug efflux and the presence of target-gene mutations, a reality that is often disregarded by the tuberculosis specialists in favor of the almost undisputed importance of antibiotic target-gene mutations for the resistance in M. tuberculosis.

BACKGROUND:Trichomoniasis is the most prevalent non-viral sexually transmitted disease (STD) in the world; however, it remains a neglected parasitic disease. This study aimed to determine the prevalence of trichomoniasis and its associated epidemiological factors among women treated at a hospital in southern Brazil. METHODOLOGY/PRINCIPAL FINDINGS:A cross-sectional study was performed to determine the prevalence of this infection in women treated at Hospital Universitário (HU) in Rio Grande, Rio Grande do Sul, Brazil, between January 2012 and January 2015. This study consisted a self-administered questionnaire regarding demographic, clinical, and behavioural data and a molecular diagnosis with polymerase chain reaction (PCR) using the TVK3/7 primer set, which was confirmed with sequence analysis. Of the 345 women surveyed, the overall prevalence of Trichomonas vaginalis (T. vaginalis) was 4.1% (14/345). The prevalence rates were 5.9% among pregnant women, 8.5% among HIV-positive women, and 10.1% among HIV-positive pregnant women. The rates for groups with other significant demographic and clinical features were as follows: 6.6% among women with white skin, 12.3% among women with an income below the minimum monthly wage, 7.4% among women with a vaginal pH greater than or equal to 4.6, and 7.9% among women with a comorbid STD. The multivariate analysis confirmed that pregnant women who were HIV-positive (p = 0.001) and had low incomes (p = 0.026) were the most likely to have this infection. CONCLUSIONS:A multivariate analysis confirmed that HIV-positive pregnant women with low incomes were the participants most likely to have trichomoniasis. These results are important because this Brazilian region presents a high prevalence of HIV-1 subtype C, which is associated with greater transmissibility. Additionally, low family income reveals a socioeconomic fragility that might favour the transmission of this STD.

The basis of drug resistance in Mycobacterium abscessus is still poorly understood. Nevertheless, as seen in other microorganisms, the efflux of antimicrobials may also play a role in M. abscessus drug resistance. Here, we investigated the role of efflux pumps in clarithromycin resistance using nine clinical isolates of M. abscessus complex belonging to the T28 erm(41) sequevar responsible for the inducible resistance to clarithromycin. The strains were characterized by drug susceptibility testing in the presence/absence of the efflux inhibitor verapamil and by genetic analysis of drug-resistance-associated genes. Efflux activity was quantified by real-time fluorometry. Efflux pump gene expression was studied by RT-qPCR upon exposure to clarithromycin. Verapamil increased the susceptibility to clarithromycin from 4- to ≥64-fold. The efflux pump genes MAB_3142 and MAB_1409 were found consistently overexpressed. The results obtained demonstrate that the T28 erm(41) polymorphism is not the sole cause of the inducible clarithromycin resistance in M. abscessus subsp. abscessus or bolletii with efflux activity providing a strong contribution to clarithromycin resistance. These data highlight the need for further studies on M. abscessus efflux response to antimicrobial stress in order to implement more effective therapeutic regimens and guidance in the development of new drugs against these bacteria.

KEYWORDS: Helicobacter pylori. Polymerase Chain Reaction. Molecular Diagnostic Techniques

Backgound and Objectives: This study is justified by the high TB prevalence in prisons, which constitutes a public health problem and aims to estimate the prevalence of active tuberculosis (TB) and determine the variables associated with respiratory symptoms in a prison in Brazil. Methods: This is a descriptive study of 262 inmates divided into respiratory symptomatic and asymptomatic groups. Samples were evaluated by microscopy following the cultivation of the sputum from symptomatic individuals. Associated epidemiological variables were also evaluated. Results: Among the 262 inmates included, 178 (68%) were considered symptomatic, and of these, 25 (14%) were diagnosed with active TB. The contribution of culturing in the detection of TB cases was 48%. The prevalence of active TB was 9,542/100.000. Low educational level, use of drugs and alcohol, prison recidivism, and previous TB and HIV-positive status were associated with the presence of respiratory symptoms. Being male, single, black, a prison recidivist, an alcoholic and HIV-seropositive was associated with the development of TB. The rate of TB/HIV co infection was 60%. The outcome was death in 12% of patients. Drug therapy interruption was reported by 96% of patients. Conclusions: The studied population showed a high prevalence of TB and TB/HIV co-infection. In addition, the rates of drug therapy interruption and mortality were alarmingly elevated. KEYWORDS: Epidemiology. Tuberculosis. Coinfections. HIV infection. Prisons.

Introduction Helicobacter pylori infection is associated with gastro-duodenal diseases. Genes related to pathogenicity have been described for H. pylori and some of them appear to be associated with more severe clinical outcomes of the infection. The present study investigates the role of cagE as a pathogenicity biomarker of H. pylori compare it to cagA, vacA, iceA and babA2 genes and correlate with endoscopic diagnoses. Methods Were collected biopsy samples of 144 dyspeptic patients at the Hospital of the Federal University of Rio Grande, Rio Grande do Sul, Brazil. After collection, the samples were sent for histological examination, DNA extraction and detection of all putative pathogenicity genes by PCR. Results Of the 144 patients undergoing endoscopy, 57 (39.6%) presented H. pylori by histological examination and PCR by detection of the ureA gene. Based on the endoscopic diagnoses, 45.6% (26/57) of the patients had erosive gastritis, while 54.4% (31/57) had enanthematous gastritis. The genes cagA, cagE, vacAs1/m1, vacAs1/m2 and iceA1 were related to erosive gastritis, while the genes vacAs2/m2, iceA2 and babA2 were associated to enanthematous gastritis. We found a statistically significant association between the presence of cagE and the endoscopic diagnosis. However, we detect no statistically significant association between the endoscopic diagnosis and the presence of cagA, vacA, iceA and babA2, although a biological association has been suggested. Conclusions Thus, cagE could be a risk biomarker for gastric lesions and may contribute to a better evaluation of the H. pylori pathogenic potential and to the prognosis of infection evolution in the gastric mucosa.

INTRODUCTION: Tuberculosis (TB) control is linked to the availability of qualified methods for microbiological diagnostics; however, microscopy with limited sensitivity is the only method available in many locations. The objective of this study was to evaluate the introduction of culture, drug susceptibility testing (DST), and genotyping in the routine of a Municipal Program of Tuberculosis Control. METHODS: Direct microscopy of sputum and culture in Ogawa-Kudoh were performed on 1,636 samples from 787 patients. DST of positive cultures was performed by resazurin microtiter assay and genotyping by mycobacterial interspersed repetitive units-variable number tandem repeat. RESULTS: A total 91 patients with TB were identified. The culture increased case detection by 32% compared with the microscopy; acquired resistance was 3.3% and the genotyping showed high genetic diversity. CONCLUSIONS: Ogawa-Kudoh contributed significantly to the increase in case detection and is suitable for implementation in poor-resource locations. The acquired resistance rate was lower than that reported in a recent Brazilian survey. The high genetic diversity is possibly related to the high TB prevalence in the population, as well as to early detection and suitable treatment of patients. The interaction between research and health care is important for reorienting the practice, transferring technology, and improving TB control.

BACKGROUND: Multidrug resistant tuberculosis (MDR-TB) is a major threat for global tuberculosis control. The W-Beijing Mycobacterium tuberculosis genotype has been associated with drug resistance. Elucidation of the mechanisms underlying this epidemiological finding may have an important role in the control of MDR-TB. The aim of this study was to evaluate the fitness of drug-susceptible and MDR M. tuberculosis strains of the W-Beijing genotype compared with that of Non-W-Beijing strains. METHODOLOGY/PRINCIPAL FINDINGS: Fitness of M. tuberculosis strains was determined by evaluating the difference in the growth curves obtained in the MGIT960 automated system and assessing the competitive growth capacity between W-Beijing and non-W-Beijing strains. The W-Beijing MDR strains had a significant longer lag phase duration compared to the other strains but did not present a significant fitness cost. When grown in competition they had an advantage only in medium containing 0.1% Tween 80. CONCLUSIONS/SIGNIFICANCE: It was not possible to confirm a selective advantage of W-Beijing strains to grow, except for differences in their resistance to Tween 80. Further studies are needed to elucidate the putative advantage of W-Beijing strains compared to other genotypes.

ABSTRACT Background Helicobacter pylori has a worldwide distribution and is associated with the pathogenesis of various diseases of the digestive system. Treatment to eradicate this microorganism involves the use of a combination of antimicrobials, such as amoxicillin, metronidazole, clarithromycin, and levofloxacin, combined with proton pump inhibitors. Although the current therapy is effective, a high rate of treatment failure has been observed, mainly because of the acquisition of point mutations, one of the major resistance mechanisms developed by H. pylori. This phenomenon is related to frequent and/or inappropriate use of antibiotics. Conclusion This review reported an overview of the resistance to the main drugs used in the treatment of H. pylori, confirming the hypothesis that antibacterial resistance is a highly local phenomenon and genetic characteristics of a given population can influence which therapy is the most appropriate.