Risk factors for extended-spectrum beta-lactamase (ESBL)-producing E. coli carriage among children in a food animal-producing region of Ecuador: A repeated measures observational study.

BACKGROUND: The spread of antibiotic-resistant bacteria may be driven by human-animal-environment interactions, especially in regions with limited restrictions on antibiotic use, widespread food animal production, and free-roaming domestic animals. In this study, we aimed to identify risk factors related to commercial food animal production, small-scale or "backyard" food animal production, domestic animal ownership, and practices related to animal handling, waste disposal, and antibiotic use in Ecuadorian communities. METHODS AND FINDINGS: We conducted a repeated measures study from 2018 to 2021 in 7 semirural parishes of Quito, Ecuador to identify determinants of third-generation cephalosporin-resistant E. coli (3GCR-EC) and extended-spectrum beta-lactamase E. coli (ESBL-EC) in children. We collected 1,699 fecal samples from 600 children and 1,871 domestic animal fecal samples from 376 of the same households at up to 5 time points per household over the 3-year study period. We used multivariable log-binomial regression models to estimate relative risks (RR) of 3GCR-EC and ESBL-EC carriage, adjusting for child sex and age, caregiver education, household wealth, and recent child antibiotic use. Risk factors for 3GCR-EC included living within 5 km of more than 5 commercial food animal operations (RR: 1.26; 95% confidence interval (CI): 1.10, 1.45; p-value: 0.001), household pig ownership (RR: 1.23; 95% CI: 1.02, 1.48; p-value: 0.030) and child pet contact (RR: 1.23; 95% CI: 1.09, 1.39; p-value: 0.001). Risk factors for ESBL-EC were dog ownership (RR: 1.35; 95% CI: 1.00, 1.83; p-value: 0.053), child pet contact (RR: 1.54; 95% CI: 1.10, 2.16; p-value: 0.012), and placing animal feces on household land/crops (RR: 1.63; 95% CI: 1.09, 2.46; p-value: 0.019). The primary limitations of this study are the use of proxy and self-reported exposure measures and the use of a single beta-lactamase drug (ceftazidime with clavulanic acid) in combination disk diffusion tests for ESBL confirmation, potentially underestimating phenotypic ESBL production among cephalosporin-resistant E. coli isolates. To improve ESBL determination, it is recommended to use 2 combination disk diffusion tests (ceftazidime with clavulanic acid and cefotaxime with clavulanic acid) for ESBL confirmatory testing. Future studies should also characterize transmission pathways by assessing antibiotic resistance in commercial food animals and environmental reservoirs. CONCLUSIONS: In this study, we observed an increase in enteric colonization of antibiotic-resistant bacteria among children with exposures to domestic animals and their waste in the household environment and children living in areas with a higher density of commercial food animal production operations.

Leveraging the COVID-19 pandemic as a natural experiment to assess changes in antibiotic use and antibiotic-resistant E. coli carriage in semi-rural Ecuador.

The coronavirus 2019 (COVID-19) pandemic has had significant impacts on health systems, population dynamics, public health awareness, and antibiotic stewardship, which could affect antibiotic resistant bacteria (ARB) emergence and transmission. In this study, we aimed to compare knowledge, attitudes, and practices (KAP) of antibiotic use and ARB carriage in Ecuadorian communities before versus after the COVID-19 pandemic began. We leveraged data collected for a repeated measures observational study of third-generation cephalosporin-resistant E. coli (3GCR-EC) carriage among children in semi-rural communities in Quito, Ecuador between July 2018 and September 2021. We included 241 households that participated in surveys and child stool sample collection in 2019, before the pandemic, and in 2021, after the pandemic began. We estimated adjusted Prevalence Ratios (aPR) and 95% Confidence Intervals (CI) using logistic and Poisson regression models. Child antibiotic use in the last 3 months declined from 17% pre-pandemic to 5% in 2021 (aPR: 0.30; 95% CI 0.15, 0.61) and 3GCR-EC carriage among children declined from 40 to 23% (aPR: 0.48; 95% CI 0.32, 0.73). Multi-drug resistance declined from 86 to 70% (aPR: 0.32; 95% CI 0.13; 0.79), the average number of antibiotic resistance genes (ARGs) per 3GCR-EC isolate declined from 9.9 to 7.8 (aPR of 0.79; 95% CI 0.65, 0.96), and the diversity of ARGs was lower in 2021. In the context of Ecuador, where COVID-19 prevention and control measures were strictly enforced after its major cities experienced some of the world's the highest mortality rates from SARS-CoV-2 infections, antibiotic use and ARB carriage declined in semi-rural communities of Quito from 2019 to 2021.

Serological testing for Hansen's disease diagnosis: Clinical significance and performance of IgA, IgM, and IgG antibodies against Mce1A protein.

Hansen's disease (HD) is an infectious, treatable, and chronic disease. It is the main cause of infectious peripheral neuropathy. Due to the current limitations of laboratory tests for the diagnosis of HD, early identification of infected contacts is an important factor that would allow us to control the magnitude of this disease in terms of world public health. Thus, a cross-sectional study was conducted in the Brazilian southeast with the objective of evaluating humoral immunity and describing the accuracy of the immunoassay based on IgA, IgM, and IgG antibodies against surface protein Mce1A of Mycobacterium, the predictive potential of these molecules, the clinical significance of positivity, and the ability to segregate new HD cases (NC; n = 200), contacts (HHC; n = 105), and healthy endemic controls (HEC; n = 100) as compared to α-PGL-I serology. α-Mce1A levels for all tested antibodies were significantly higher in NC and HHC than in HEC (p < 0.0001). The performance of the assay using IgA and IgM antibodies was rated as highly accurate (AUC > 0.85) for screening HD patients. Among HD patients (NC), positivity was 77.5% for IgA α-Mce1A ELISA, 76.5% for IgM, and 61.5% for IgG, while α-PGL-I serology showed only 28.0% positivity. Multivariate PLS-DA showed two defined clusters for the HEC and NC groups [accuracy = 0.95 (SD = 0.008)] and the HEC and HHC groups [accuracy = 0.93 (SD = 0.011)]. IgA was the antibody most responsible for clustering HHC as compared to NC and HEC, evidencing its usefulness for host mucosal immunity and as an immunological marker in laboratory tests. IgM is the key antibody for the clustering of NC patients. Positive results with high antibody levels indicate priority for screening, new clinical and laboratory evaluations, and monitoring of contacts, mainly with antibody indexes ≥2.0. In light of recent developments, the incorporation of new diagnostic technologies permits to eliminate the main gaps in the laboratory diagnosis of HD, with the implementation of tools of greater sensitivity and accuracy while maintaining satisfactory specificity.

Evaluation of changes in antimicrobial susceptibility in bacteria infecting children and their mothers in pediatric, neonatal-intensive care unit, and gynecology/obstetrics wards of a quaternary referral hospital during the COVID-19 pandemic.

The World Health Organization released a statement warning of increased risk for the incidence of multidrug resistant microorganisms and the absence of new drugs to control such infections soon. Since the beginning of the COVID-19 pandemic, the prescription of antimicrobial agents has increased and may have accelerated the emergence of multidrug resistant (MDR) bacteria. This study aimed to evaluate maternal and pediatric infections within a hospital from January 2019 to December 2021. An observational retrospective cohort study was performed at a quaternary referral hospital in a metropolitan area of Niteroi city, Rio de Janeiro state, Brazil. A total of 196 patients' medical records were analyzed. The data from 90 (45.9%) patients were collected before the SARS-CoV-2 pandemic, 29 (14.8%) from the 2020 pandemic period, and 77 (39.3%) from the 2021 pandemic period. A total of 256 microorganisms were identified during this period. Out of those, 101 (39.5%) were isolated in 2019, 51 (19.9%) in 2020, and 104 (40.6%) in 2021. Antimicrobial susceptibility tests were performed on 196 (76.6%) clinical isolates. The exact binomial test showed that the distribution of Gram-negative bacteria was predominant. The most common microorganism was Escherichia coli (23%; n = 45), followed by Staphylococcus aureus (17.9%, n = 35), Klebsiella pneumoniae (12.8%, n = 25), Enterococcus faecalis (7.7%, n = 15), Staphylococcus epidermidis (6.6%, n = 13) and Pseudomonas aeruginosa (5.6%, n = 11). Staphylococcus aureus was the predominant species among resistant bacteria. Among the antimicrobial agents tested, the following were resistant, presented on a descending scale: penicillin (72.7%, p = 0.001, Binomial test), oxacillin (68.3%, p = 0.006, Binomial test), ampicillin (64.3%, p = 0.003, Binomial test), and ampicillin/sulbactam (54.9%, p = 0.57, Binomial test). Infections with S. aureus were 3.1 times greater in pediatrics and maternal units than in other hospital wards. Despite the global reduction in the incidence of MRSA, we observed an increase in MDR S. aureus in this study. No changes were observed in the frequency of resistance profiles of the clinical isolates after the establishment of the global SARS-CoV-2 pandemic. More comprehensive studies are needed to understand the impact of the global SARS-CoV-2 pandemic on the resistance levels of bacteria associated with neonate and pediatric patients.

A Matter of Inclusion: A Cluster-Randomized Trial to Access the Effect of Food Vouchers Versus Traditional Treatment on Tuberculosis Outcomes in Brazil.

We assessed the effectiveness of food vouchers as a social protection strategy to enhance the adherence to tuberculosis treatment in health-care facilities in Brazil between 2014 and 2017. A cluster-randomized controlled trial was performed in four Brazilian capital cities. A total of 774 adults with newly diagnosed pulmonary tuberculosis were included in this study. Eligible participants initiated standard tuberculosis treatment per National Tuberculosis Program guidelines. Health clinics were assigned randomly to intervention groups (food voucher or standard treatment). The food voucher was provided by researchers, which could be used by subjects only for buying food. Most people with tuberculosis were poor, did receive benefits of the Bolsa Familia Program, and were unemployed. By Poisson regression analysis, with the total number of subjects included in the study, we found that individuals with tuberculosis who received food vouchers had a 1.13 greater risk of cure (95% CI, 1.03-1.21) compared with those who did not receive food vouchers. The provision of food vouchers improved outcomes of tuberculosis treatment and it should be enhanced even further as social protection for people with tuberculosis.

Serological Immunoassay for Hansen's Disease Diagnosis and Monitoring Treatment: Anti-Mce1A Antibody Response Among Hansen's Disease Patients and Their Household Contacts in Northeastern Brazil.

Hansen's disease (HD) is an ancient disease, but more than 200,000 new cases were reported worldwide in 2019. Currently, there are not many satisfactory immunoassay methods for its diagnosis. We evaluated antibodies against Mce1A as a promising new serological biomarker. We collected plasma from new cases, contacts, and endemic controls in the city of Parnaíba and treated patients at Carpina, a former HD colony in Piauí state, northeastern Brazil. Receiver operating characteristic (ROC) curves were used to assess the assay thresholds, specificity and sensitivity of the IgA, IgM, and IgG antibodies against α-Mce1A by indirect ELISA and compared it with IgM anti-PGL-I and molecular diagnosis by quantitative polymerase chain reaction (qPCR). Venn diagrams were generated to represent the overlap in the antibody positivity pattern. Multivariate analysis was performed to assess the potential predictor of antibodies for the outcome of having an HD diagnosis. IgA and IgG were positive in 92.3 and 84% of patients, respectively. IgM was negative for all treated patients. IgG had a sensitivity and specificity of 94.7 and 100%, respectively. IgM-positive individuals had a 3.6 chance of being diagnosed with HD [OR = 3.6 (95% CI = 1.1-11.6); p = 0.028], while IgA-positive individuals had a 2.3 chance [OR = 2.3 (95% CI = 1.2-4.3); p = 0.005] compared to endemic controls. We found that the Mce1A antibody profile can be an excellent diagnostic method of HD. IgA is an ideal biomarker for confirming contact with the bacillus. IgM has potential in the detection of active disease. IgG antibodies confirm the performance of these serological markers in diagnosis and therapeutic follow-up.

Whole blood mRNA expression-based targets to discriminate active tuberculosis from latent infection and other pulmonary diseases.

Current diagnostic tests for tuberculosis (TB) are not able to predict reactivation disease progression from latent TB infection (LTBI). The main barrier to predicting reactivation disease is the lack of our understanding of host biomarkers associated with progression from latent infection to active disease. Here, we applied an immune-based gene expression profile by NanoString platform to identify whole blood markers that can distinguish active TB from other lung diseases (OPD), and that could be further evaluated as a reactivation TB predictor. Among 23 candidate genes that differentiated patients with active TB from those with OPD, nine genes (CD274, CEACAM1, CR1, FCGR1A/B, IFITM1, IRAK3, LILRA6, MAPK14, PDCD1LG2) demonstrated sensitivity and specificity of 100%. Seven genes (C1QB, C2, CCR2, CCRL2, LILRB4, MAPK14, MSR1) distinguished TB from LTBI with sensitivity and specificity between 82 and 100%. This study identified single gene candidates that distinguished TB from OPD and LTBI with high sensitivity and specificity (both > 82%), which may be further evaluated as diagnostic for disease and as predictive markers for reactivation TB.

Differential Host Pro-Inflammatory Response to Mycobacterial Cell Wall Lipids Regulated by the Mce1 Operon.

The cell wall of wild-type (WT) Mycobacterium tuberculosis (Mtb), an etiologic agent of tuberculosis (TB) and a Mtb strain disrupted in a 13-gene operon mce1 (Δmce1) varies by more than 400 lipid species. Here, we examined Mtb lipid-induced response in murine macrophage, as well as in human T-cell subpopulations in order to gain an insight into how changes in cell wall lipid composition may modulate host immune response. Relative to WT Mtb cell wall lipids, the non-polar lipid extracts from Δmce1 enhanced the mRNA expression of lipid-sense nuclear receptors TR4 and PPAR-γ and dampened the macrophage expression of genes encoding TNF-α, IL-6, and IL-1ß. Relative to untreated control, WT lipid-pre-stimulated macrophages from healthy individuals induced a higher level of CD4-CD8- double negative T-cells (DN T-cells) producing TNF-α. Conversely, compared to WT, stimulation with Δmce1 lipids induced higher mean fluorescence intensity (MFI) in IL-10-producing DN T cells. Mononuclear cells from TB patients stimulated with WT Mtb lipids induced an increased production of TNF-α by CD8+ lymphocytes. Taken together, these observations suggest that changes in mce1 operon expression during a course of infection may serve as a strategy by Mtb to evade the host pro-inflammatory responses.

Serological biomarkers for monitoring response to treatment of pulmonary and extrapulmonary tuberculosis in children and adolescents.

Key measures to halt the spread of tuberculosis (TB) include early diagnosis, effective treatment, and monitoring disease management. We sought to evaluate the use of serum immunoglobulin levels against antigens present in cell envelope of Mycobacterium tuberculosis to monitor TB treatment response in children and adolescents with pulmonary (PTB) or extrapulmonary TB (EPTB). Blood samples were collected prior to and one, two, and six months following treatment initiation. Serum immunoglobulin levels against cardiolipin, sulfatide, mycolic acid and Mce1A protein were measured by ELISA. Serum from 53 TB patients and 12 healthy participants were analyzed. After six months of successful treatment, there was a significant decrease (p < 0.0001) in IgM levels against cardiolipin, sulfatide, mycolic acid and Mce1A protein and IgG levels against Mce1A protein when compared to baseline immunoglobulin levels. There was no significant variation in antibody levels during follow-up between participants with PTB and EPTB, confirmed and unconfirmed TB diagnosis, and HIV infection status. Antibody levels in control participants without TB did not decrease during follow-up. These results suggest that immunoglobulin responses to mycobacterial cell wall products may be a useful tool to monitor treatment response in children and adolescents with PTB or EPTB.

Genotypic distribution of Staphylococcus aureus colonizing children and adolescents in daycare centers, an outpatient clinic, and hospitals in a major Brazilian urban setting.

Staphylococcus aureus nasal colonization is a major risk factor for infection. Studies have suggested an epidemiologic shift in the methicillin-resistant S. aureus (MRSA) strains that circulate in Brazil. We conducted cross-sectional studies of MRSA carriage among 1) children and adolescents in community daycare centers, 2) an outpatient clinic, and 3) hospitals in a large Brazilian metropolitan setting. There were 1.500 study subjects, 500 from each locale: 768 (51.2%) carried S. aureus whereas 150 (10%) of these were colonized with MRSA. The most common lineages were the Southwest Pacific (SWP) and the Pediatric clones in all three groups. Roughly 50% of SWP carried Panton-Valentine leukocidin (PVL) (p < 0.01) genes while 63.3% of the Pediatric clones were resistant or intermediately resistant to erythromycin (p < 0.01). This study describes a clonal change of the Brazilian epidemic clone (BEC) to the Pediatric and SWP lineages in Brazil. This finding has implications for clinical management of MRSA infections.

A novel plasmid-encoded mcr-4.3 gene in a colistin-resistant Acinetobacter baumannii clinical strain.

OBJECTIVES: To identify the molecular mechanism of colistin resistance in an MDR Acinetobacter baumannii clinical strain isolated in 2008 from a meningitis case in Brazil. METHODS: Long- and short-read WGS was used to identify colistin resistance genes in A. baumannii strain 597A with a colistin MIC of 64 mg/L. MS was used to analyse lipid A content. mcr was cloned into pET-26b (+) and transformed into Escherichia coli BL21(λDE3)pLysS for analysis. RESULTS: A novel plasmid (pAb-MCR4.3) harbouring mcr-4.3 within a Tn3-like transposon was identified. The A. baumannii 597A lipid A MS spectra showed a main molecular ion peak at m/z=2034, which indicated the addition of phosphoethanolamine to the lipid A structure. E. coli BL21 transformed with pET-26b-mcr-4.3 gained colistin resistance with a colistin MIC of 8 mg/L. CONCLUSIONS: Colistin resistance in A. baumannii 597A was correlated with the presence of a novel plasmid-encoded mcr-4.3 gene.

[Variations in the detection of congenital Zika syndrome associated with changes in protocolsVariación de la detección del síndrome congénito por el virus del Zika en función de los cambios de los protocolos]. / Variação na detecção da síndrome congênita do Zika em função de alterações em protocolos.

In 2015, Brazil faced a Zika virus epidemic that spread to other countries in the world. As a result, recommendations regarding reporting criteria for congenital Zika syndrome (CZS) were issued in the form of protocols. The frequent changes in these recommendations may have affected clinical management and the access to post-diagnostic support by children who were affected by CZS, but who ended up not being identified. In the present study, 39 cases of CZS reported in the state of Espírito Santo, Brazil, from 2015 to 2016 were re-classified in terms of diagnosis using the current protocol, which is different from the protocol used in 2015. According to this re-classification, only eight out of 36 cases would be confirmed, based on the criterion of two or more signs or symptoms of CZS with or without microcephaly plus positive serologic results. Also, considering the decrease in the head circumference cut-off point defining microcephaly, 17 cases would no longer meet the definition for this condition. Even though the current protocol does not rely on head circumference alone for CZS reporting and confirmation, it should be noted that this is still the main sign considered by health care teams, and therefore the decrease in the cut-off point might have compromised early CZS detection. A review of "ruled out" cases would be advisable in moments of protocol transition to determine whether these cases have been correctly classified.

En el 2015, Brasil enfrentó una epidemia de infección por el virus del Zika que se propagó por varios países del mundo. Posteriormente, se divulgaron recomendaciones acerca de los criterios de notificación de casos del síndrome congénito por el virus del Zika (SCZ) por medio de protocolos. Los cambios frecuentes de esas recomendaciones podrían haber afectado el manejo clínico y el acceso al apoyo posterior al diagnóstico de los niños afectados, pero no identificados. En el presente estudio, se reclasificó el diagnóstico de 39 casos del SCZ notificados en el estado de Espírito Santo en el período 2015-2016, de acuerdo con el protocolo vigente en la actualidad, que es distinto del que regía en el 2015. Por causa de la reclasificación, se confirmaron únicamente ocho de los 36 casos, con observancia del criterio de dos o más signos o síntomas del SCZ acompañados o no de microcefalia y con confirmación serológica. Además, por la disminución del perímetro cefálico que define la microcefalia, 17 casos no correspondieron a esa afección. A pesar de que en el protocolo vigente no se utiliza solamente el perímetro cefálico como criterio para la notificación y confirmación del SCZ, cabe resaltar que este hallazgo es, con todo, la mayor señal para los equipos de salud, puesto que indica un riesgo de falta de detección temprana del SCZ. Convendría examinar los casos "descartados" en el momento de la transición entre protocolos, con el fin de determinar si se clasificaron correctamente.

Variação na detecção da síndrome congênita do Zika em função de alterações em protocolos

[RESUMO]. Em 2015, o Brasil enfrentou uma epidemia de infecção pelo vírus Zika que se propagou por países do mundo. Posteriormente, recomendações acerca dos critérios de notificação de casos de síndrome congênita do Zika (SCZ) foram divulgadas através de protocolos. As mudanças frequentes nessas recomendações podem ter afetado o gerenciamento clínico e o acesso a suporte pós-diagnóstico por crianças afetadas mas não identificadas. No presente estudo, 39 casos de SCZ notificados no estado do Espírito Santo no período de 2015 a 2016 foram reclassificados quanto ao seu diagnóstico de acordo com o protocolo atualmente vigente, diferente daquele que vigorava em 2015. Pela reclassificação, apenas oito dos 36 casos seriam confirmados, respeitando o critério de dois ou mais sinais ou sintomas da SCZ com ou sem microcefalia e confirmação sorológica. Ainda, pela diminuição no perímetro cefálico que define microcefalia, 17 casos passaram a não se enquadrar nessa condição. Apesar de o protocolo vigente não utilizar somente o perímetro cefálico como critério para notificação e confirmação da SCZ, cabe ressaltar que este achado ainda é o maior sinalizador para as equipes de saúde, indicando um risco da não detecção precoce da SCZ. Seria prudente uma revisão dos casos “descartados” no momento de transição entre protocolos, a fim de avaliar se foram corretamente classificados.

[ABSTRACT]. In 2015, Brazil faced a Zika virus epidemic that spread to other countries in the world. As a result, recommendations regarding reporting criteria for congenital Zika syndrome (CZS) were issued in the form of protocols. The frequent changes in these recommendations may have affected clinical management and the access to post-diagnostic support by children who were affected by CZS, but who ended up not being identified. In the present study, 39 cases of CZS reported in the state of Espírito Santo, Brazil, from 2015 to 2016 were re-classified in terms of diagnosis using the current protocol, which is different from the protocol used in 2015. According to this re-classification, only eight out of 36 cases would be confirmed, based on the criterion of two or more signs or symptoms of CZS with or without microcephaly plus positive serologic results. Also, considering the decrease in the head circumference cut-off point defining microcephaly, 17 cases would no longer meet the definition for this condition. Even though the current protocol does not rely on head circumference alone for CZS reporting and confirmation, it should be noted that this is still the main sign considered by health care teams, and therefore the decrease in the cut-off point might have compromised early CZS detection. A review of “ruled out” cases would be advisable in moments of protocol transition to determine whether these cases have been correctly classified.

[RESUMEN]. En el 2015, Brasil enfrentó una epidemia de infección por el virus del Zika que se propagó por varios países del mundo. Posteriormente, se divulgaron recomendaciones acerca de los criterios de notificación de casos del síndrome congénito por el virus del Zika (SCZ) por medio de protocolos. Los cambios frecuentes de esas recomendaciones podrían haber afectado el manejo clínico y el acceso al apoyo posterior al diagnóstico de los niños afectados, pero no identificados. En el presente estudio, se reclasificó el diagnóstico de 39 casos del SCZ notificados en el estado de Espírito Santo en el período 2015-2016, de acuerdo con el protocolo vigente en la actualidad, que es distinto del que regía en el 2015. Por causa de la reclasificación, se confirmaron únicamente ocho de los 36 casos, con observancia del criterio de dos o más signos o síntomas del SCZ acompañados o no de microcefalia y con confirmación serológica. Además, por la disminución del perímetro cefálico que define la microcefalia, 17 casos no correspondieron a esa afección. A pesar de que en el protocolo vigente no se utiliza solamente el perímetro cefálico como criterio para la notificación y confirmación del SCZ, cabe resaltar que este hallazgo es, con todo, la mayor señal para los equipos de salud, puesto que indica un riesgo de falta de detección temprana del SCZ. Convendría examinar los casos “descartados” en el momento de la transición entre protocolos, con el fin de determinar si se clasificaron correctamente.

Direct effect of the 13-valent pneumococcal conjugate vaccine use on pneumococcal colonization among children in Brazil.

BACKGROUND: The 13-valent pneumococcal conjugate vaccine (PCV13) has been commercially available in Brazil since 2010. We investigated the carriage prevalence, capsular types, and antimicrobial resistance among pneumococci isolated from children immunized with PCV13 in Brazil. METHODS: We analyzed 500 children < 6 years old attending public (n = 270) and private (n = 230) clinics in Niterói/RJ, Brazil, in 2014. We determined the antimicrobial susceptibility and capsular types for all isolates. RESULTS: Thirty-eight (7.6%) of 500 children had received at least one PCV13 dose. Since only two (0.7%) of 270 children at the public clinic were vaccinated with PCV13, major analyses focused on 36 (15.7%) of 230 children attending private clinics. Nine (25%) of 36 children were pneumococcal carriers. Characteristics associated with carriage were age ≥ 2 years, cough/expectoration, and childcare center attendance (p ≤ 0.01). The capsular types found were 15B/C (n = 2), 6C, 11A/D, 16F, 23A, and 23F. Two isolates were non-typeable (NT). Three (33.3%) isolates were multidrug resistant. We found four (44.4%) penicillin non-susceptible pneumococci, with penicillin and ceftriaxone MICs ranging from 0.12 to 4.0 µg/ml and 0.023-0.5 µg/ml, respectively. We also detected two (22.2%) erythromycin-resistant isolates (MICs of 3.0 and 256 µg/ml). CONCLUSIONS: Colonization with PCV13 serotype was rare among the vaccinated children. Increasing PCV13 coverage might help reduce the frequency of major serotypes currently associated with invasive pneumococcal diseases in Brazil, such as 3 and 19A. The isolation of multidrug-resistant serotype 6C and NT isolates in carriage, however, requires close monitoring.

Effect of the Bolsa Familia Programme on the outcome of tuberculosis treatment: a prospective cohort study.

BACKGROUND: Social protection interventions might improve tuberculosis outcomes and could help to control the epidemic in Brazil. The aim of this study was to evaluate the independent effect of the Bolsa Familia Programme (BFP) on tuberculosis treatment outcomes in Brazil. METHODS: We prospectively recruited and followed up individuals (aged ≥18 years) who initiated tuberculosis treatment at 42 health-care centres across seven cities in Brazil, between March 1, 2014, and April 30, 2017. Patients were interviewed at health-care centres and information about individual characteristics, socioeconomic status, living conditions, lifestyle, and comorbidities was recorded. Patients were separated into two groups according to BFP beneficiary status: BFP (exposed) or non-BFP (not exposed). Treatment outcome (cure, dropout, death, or development of drug-resistant tuberculosis or treatment failure) was recorded after 6 months of therapy. Pearson's χ2 test and ANOVA were used to compare tuberculosis treatment outcomes between the two groups, and we estimated the propensity score of being a beneficiary of the BFP using a logit model. We used multinomial regression models to evaluate the effect of the BFP on tuberculosis treatment outcomes. FINDINGS: 1239 individuals were included in the study, of whom 196 (16%) were beneficiaries of the BFP and 1043 (84%) were not. After 6 months of treatment, 912 (87%) of 1043 patients in the non-BFP group and 173 (88%) of 196 patients in the BFP group were cured of tuberculosis, 103 (10%) patients in the non-BFP group and 17 (9%) patients in the BFP group had dropped out, and 25 (3%) patients in the non-BFP group and six (3%) patients in the BFP group had died. Three (<1%) of 1043 patients in the non-BFP group developed drug-resistant tuberculosis. Being a BFP beneficiary had a positive effect for cure (average effect 0·076 [95% CI 0·037 to 0·11]) and a negative effect for dropout (-0·070 [-0·105 to 0·036]) and death (-0·002 [-0·021 to 0·017]). INTERPRETATION: BFP alone had a direct effect on tuberculosis treatment outcome and could greatly contribute to the goals of the WHO End TB Strategy. FUNDING: Brazilian National Council for Scientific and Technological Development (CNPq) and Brazilian Ministry of Health Department of Science and Technology (DECIT).

Population structure of Streptococcus pneumoniae colonizing children before and after universal use of pneumococcal conjugate vaccines in Brazil: emergence and expansion of the MDR serotype 6C-CC386 lineage.

Objectives: To determine the population structure and change in drug resistance of pneumococci colonizing children before and after the introduction of the 10-valent and 13-valent pneumococcal conjugate vaccines (PCV10/13) in Brazil. Methods: We used MLST to analyse 256 pneumococcal isolates obtained from children aged <6 years before (2009-10; n = 125) and after (2014; n = 131) the introduction of the PCV10 and PCV13. Antimicrobial susceptibility and capsular types were previously determined. Results: We identified 97 different STs. Ninety (35.2%) isolates were related to international clones. The most frequent lineages were serogroup 6-CC724 (where CC stands for clonal complex) and the MDR serotype 6C-CC386 in the pre- and post-PCV10/13 periods, respectively. Penicillin-non-susceptible pneumococci (PNSP) formed 24% and 38.9% of the pre- and post-PCV10/13 isolates, respectively (P = 0.01). In the pre-PCV10/13 period, serotype 14-ST156 was the predominant penicillin-non-susceptible lineage, but it was not detected in the post-PCV10/13 period. Serotype 14-ST156 and serotype 19A-ST320 complex isolates had the highest penicillin and ceftriaxone MICs in the pre- and post-PCV10/13 periods, respectively. In turn, serotype 6C-CC386 comprised almost 30% of the PNSP and over 40% of the erythromycin-resistant isolates (MIC >256 mg/L) in the post-PCV10/13 period. Conclusions: Although PNSP strains were polyclonal, most resistant isolates belonged to a single genotype from each period. Higher erythromycin resistance prevalence (42%) in the post-PCV10/13 period was mainly attributed to MDR serotype 6C-CC386. Ongoing surveillance of pneumococcal clonal composition is important to evaluate PCV use outcomes and to identify factors other than PCVs that drive pneumococcal drug resistance evolution.

Risk factors associated with cluster size of Mycobacterium tuberculosis (Mtb) of different RFLP lineages in Brazil.

BACKGROUND: Tuberculosis (TB) transmission is influenced by patient-related risk, environment and bacteriological factors. We determined the risk factors associated with cluster size of IS6110 RFLP based genotypes of Mycobacterium tuberculosis (Mtb) isolates from Vitoria, Espirito Santo, Brazil. METHODS: Cross-sectional study of new TB cases identified in the metropolitan area of Vitoria, Brazil between 2000 and 2010. Mtb isolates were genotyped by the IS6110 RFLP, spoligotyping and RDRio. The isolates were classified according to genotype cluster sizes by three genotyping methods and associated patient epidemiologic characteristics. Regression Model was performed to identify factors associated with cluster size. RESULTS: Among 959 Mtb isolates, 461 (48%) cases had an isolate that belonged to an RFLP cluster, and six clusters with ten or more isolates were identified. Of the isolates spoligotyped, 448 (52%) were classified as LAM and 412 (48%) as non-LAM. Our regression model found that 6-9 isolates/RFLP cluster were more likely belong to the LAM family, having the RDRio genotype and to be smear-positive (adjusted OR = 1.17, 95% CI 1.08-1.26; adjusted OR = 1.25, 95% CI 1.14-1.37; crude OR = 2.68, 95% IC 1.13-6.34; respectively) and living in a Serra city neighborhood decrease the risk of being in the 6-9 isolates/RFLP cluster (adjusted OR = 0.29, 95% CI, 0.10-0.84), than in the others groups. Individuals aged 21 to 30, 31 to 40 and > 50 years were less likely of belonging the 2-5 isolates/RFLP cluster than unique patterns compared to individuals < 20 years of age (adjusted OR = 0.49, 95% CI 0.28-0.85, OR = 0.43 95% CI 0.24-0.77and OR = 0. 49, 95% CI 0.26-0.91), respectively. The extrapulmonary disease was less likely to occur in those infected with strains in the 2-5 isolates/cluster group (adjustment OR = 0.45, 95% CI 0.24-0.85) than unique patterns. CONCLUSIONS: We found that a large proportion of new TB infections in Vitoria is caused by prevalent Mtb genotypes belonging to the LAM family and RDRio genotypes. Such information demonstrates that some genotypes are more likely to cause recent transmission. Targeting interventions such as screening in specific areas and social risk groups, should be a priority for reducing transmission.

Prevalence and risk factors for latent tuberculosis infection among primary health care workers in Brazil.

Health care workers (HCW) are at increased risk of latent tuberculosis infection (LTBI) from occupational exposure to Mycobacterium tuberculosis. The objective was to determine the prevalence of and risk factors for LTBI among primary HCW in five Brazilian cities. We conducted a cross-sectional study, from 2011 to 2013, among primary HCW, using a structured questionnaire and an evaluated for LTBI using the Quantiferon-TB Gold in-tube test. The magnitude of the associations was assessed using hierarchical logistic regression models. Among 708 HCW, the LTBI prevalence was 27% (n = 196; 95%CI: 24%-31%). We found that the following factors were positively associated with LTBI in primary HCW: age > 50 years (OR = 2.94; 95%CI: 1.44-5.99), absence of a BCG scar (OR = 2.10; 95%CI: 1.28-3.43), self-reported ex-smoker status (OR = 1.80; 95%CI: 1.04-3.11), being a nurse (OR = 2.97; 95%CI: 1.13-7.83), being a nurse technician (OR = 3.10; 95%CI: 1.26-7.60), being a community health agent (OR = 2.60; 95%CI: 1.06-6.40), and irregular use of N95 masks (OR = 2.51; 95%CI: 1.11-5.98). In contrast, HCWs who do not work in health care facilities with a TB control program were less likely to have LTBI (OR = 0.66; 95%CI: 0.45-0.97). This study demonstrated a substantial occupational risk of LTBI among primary HCW in Brazil. The Brazilian TB control program, as well as local programs, need to target these high-risk HCW with education, as well as with better personal protective equipment to prevent acquisition of new TB infection.

ELISA-based assay of immunoglobulin G antibodies against mammalian cell entry 1A (Mce1A) protein: a novel diagnostic approach for leprosy.

BACKGROUND: Leprosy is a chronic infectious disease caused by the obligate intracellular bacillus Mycobacterium leprae. Because leprosy diagnosis is complex and requires professional expertise, new tools and methodologies are needed to detect cases in early stages and prevent transmission. The M. leprae genome contains mce1A, which encodes a putative mammalian cell entry protein (Mce1A). We hypothesised that the presence of Mce1A on the cell surface could be detected by the host's immune system. OBJECTIVE: The aim of this study was to evaluate antibody responses against the Mce1A protein in leprosy patients, household contacts of patients, and the general population to present an addition tool for leprosy diagnosis. METHODS: A cross-sectional study involving 89 volunteers [55 leprosy cases, 12 household contacts (HHC) and 22 endemic controls (EC)] was conducted at Couto Maia Hospital, in Salvador, Bahia (BA), Brazil. RESULTS: The median anti-Mce1A IgA was significantly higher in multibacillary (MB) and paucibacillary (PB) cases than in EC (p < 0.0001). A similar trend was observed in IgM levels, which were significantly higher in both MB (p < 0.0001) and PB (p = 0.0006) groups compared to in EC individuals. The greatest differences were observed for IgG class-specific antibodies against Mce1A. The median levels of MB and PB were significantly higher compared to both controls HHC and EC (MB or PB vs EC, MB vs HHC p < 0.0001; PB vs HHC, p = 0.0013). Among leprosy cases, IgG enzyme-linked immunosorbent assay sensitivity and specificity were 92.7% and 97.1%, respectively. IgG positivity was confirmed in 92.1% and 94.1% of MB and PB patients, respectively. CONCLUSION: This novel diagnostic approach presents an easy, non-invasive, and inexpensive method for leprosy screening, which may be applicable in endemic areas.