Characterization by MALDI-TOF MS and 16S rRNA Gene Sequencing of Aerobic Endospore-Forming Bacteria Isolated from Pharmaceutical Facility in Rio de Janeiro, Brazil.

Bacillus and related genera are among the most important contaminants in the pharmaceutical production environment, and the identification of these microorganisms at the species level assists in the investigation of sources of contamination and in preventive and corrective decision making. The aim of this study was to evaluate three methodologies for the characterization of endospore-forming aerobic bacterial strains isolated from a pharmaceutical unit in Rio de Janeiro, Brazil. MALDI-TOF MS was performed using MALDI Biotyper® and VITEK® MS RUO systems, and complete 16S rRNA gene sequencing was performed using the Sanger methodology. The results showed the prevalence of the genera Bacillus (n = 9; 36.0%), Priestia (n = 5; 20.0%), and Paenibacillus (n = 4; 16.0%). Three (20.0%) strains showed <98.7% of DNA sequencing similarity on the EzBioCloud Database, indicating possible new species. In addition, the reclassification of Bacillus pseudoflexus to the genus Priestia as Priestia pseudoflexus sp. nov. is proposed. In conclusion, 16S rRNA and MALDI TOF/MS were not sufficient to identify all strains at the species level, and complementary analyses were necessary.

Environmental assessment of sewage contamination in the surroundings of a marine outfall combining human mastadenovirus and fecal indicator bacteria.

This study assessed the microbiological contamination of the marine area of a metropolitan region, where a marine outfall is used as a sanitary solution for domestic sewage. For human mastadenovirus (HAdV) quantification 134 water samples were concentrated by skimmed milk flocculation method and analyzed with qPCR and PMAxx-qPCR, being the latter to assess the capsid integrity viral. HAdV with intact capsids were detected in 10 % (16/102) of samples classified as suitable for bathing using at least one fecal bacterial indicator. Spatial analysis of the results showed that the drainage channels of the basin that flow into the sea are the main sources of microbiological contamination in the foreshore zone, where intact HAdV reached a concentration of up to 3 log genomic copies/L. HAdV serotypes A12, D, F40 and F41 were characterized. Our results suggest the use of intact HAdV as a complementary parameter to assess the quality of recreational waters.

Molecular Epidemiology of Haemophilus influenzae Strains with Reduced Susceptibility and Genetic Profiles of Resistance in the Postvaccination Period.

Haemophilus influenzae serotype b has been the main cause of invasive infections in children, during the prevaccination period. More than 20 years after the introduction of the conjugate vaccine against Hib, HiNT has emerged as the cause of localized infections in children and adults. The main objective of this work is to evaluate the susceptibility and resistance mechanisms of H. influenzae strains from carriers and describe the molecular epidemiology and their clonal relationships by multilocus sequence typing (MLST). Sixty-nine strains from clinical cases and asymptomatic carriers from 2009 to 2019 were analyzed, confirmed as H. influenzae, and serotyped by polymerase chain reaction. The susceptibility to antibiotics was evaluated by E-test strips. Genotyping was performed by MLST. HiNT was the most frequent in all age groups. Resistance to ampicillin, sulfamethoxazole+trimethoprim, and amoxicillin+clavulanic acid was detected, with the production of ß-lactamase being the main resistance mechanism. Among 21 HiNT strains with complete allelic MLST profiles, 19 new sequence types were described, reinforcing the already reported heterogeneity of nontypeable strains, and only one clonal complex (cc-1355) was observed. Our results show a high percentage of colonization regardless of age, increased antimicrobial resistance, and high genetic diversity, along with an increased number of cases caused by HiNT strains. These findings reinforce the need for continuous surveillance for HiNT strains as it has been reported worldwide after the introduction of the Hib conjugate vaccine.

Breastfeeding self-efficacy and postpartum depression: a cohort study.

OBJECTIVE: to evaluate breastfeeding self-efficacy, the presence of postpartum depression symptons and the association between breastfeeding self-efficacy and postpartum depression with cessation of exclusive breastfeeding. METHOD: cohort study with 83 women. The instruments used were the Breastfeeding Self-Efficacy Scale and the Edinburgh Postnatal Depression Scale. Statistical analysis was conducted using the log-rank tests, analysis of variance and the Cox survival model. RESULTS: breastfeeding self-efficacy (p = 0.315) and postpartum depression (p = 0.0879) did not show any statistical difference over time. The chances of cessation of exclusive breastfeeding decreased by 48% when self-efficacy changed from low to medium and by 80% when it changed from medium to high. Postpartum women who scored ≥10 on the Edinburgh Postnatal Depression Scale interrupt exclusive breastfeeding, on average, 10 days earlier than those with a score ≤9, whose median breastfeeding duration was 38 days postpartum. CONCLUSION: breastfeeding self-efficacy was proved to be a protective factor for exclusive breastfeeding, while postpartum depression is a risk factor.

Breastfeeding self-efficacy and postpartum depression: a cohort study / Autoeficácia para amamentação e depressão pós-parto: estudo de coorte / Autoeficacia para el amamantamiento y depresión post-parto: estudio de cohorte

ABSTRACT Objective: to evaluate breastfeeding self-efficacy, the presence of postpartum depression symptons and the association between breastfeeding self-efficacy and postpartum depression with cessation of exclusive breastfeeding. Method: cohort study with 83 women. The instruments used were the Breastfeeding Self-Efficacy Scale and the Edinburgh Postnatal Depression Scale. Statistical analysis was conducted using the log-rank tests, analysis of variance and the Cox survival model. Results: breastfeeding self-efficacy (p = 0.315) and postpartum depression (p = 0.0879) did not show any statistical difference over time. The chances of cessation of exclusive breastfeeding decreased by 48% when self-efficacy changed from low to medium and by 80% when it changed from medium to high. Postpartum women who scored ≥10 on the Edinburgh Postnatal Depression Scale interrupt exclusive breastfeeding, on average, 10 days earlier than those with a score ≤9, whose median breastfeeding duration was 38 days postpartum. Conclusion: breastfeeding self-efficacy was proved to be a protective factor for exclusive breastfeeding, while postpartum depression is a risk factor.

RESUMO Objetivo: avaliar a autoeficácia para amamentação, a presença de sintomas de depressão no período pós-parto e a associação entre autoeficácia na amamentação e depressão pós-parto, com a interrupção do aleitamento materno exclusivo. Método: estudo de coorte realizado com 83 mulheres. Os instrumentos utilizados foram a Escala de Autoeficácia para amamentação e a Escala de Depressão Pós-natal de Edimburgo. A análise estatística foi feita por meio dos testes log-rank, análise de variância e modelo de sobrevivência de Cox. Resultados: os níveis de autoeficácia para amamentação (p=0,315) e depressão pós-parto (p=0,0879), ao longo do tempo, não evidenciaram diferença estatística. As chances de interrupção do aleitamento materno exclusivo diminuem em 48% com a melhora do nível baixo de autoeficácia para médio e em 80% de médio para alto. Puérperas com escore ≥10 na Escala de Depressão Pós-natal de Edimburgo interrompem o aleitamento exclusivo, em média, 10 dias antes do que aquelas com escore ≤9, cuja mediana de aleitamento materno foi de 38 dias após o parto. Conclusão: a autoeficácia para amamentação revelou-se como fator de proteção para o aleitamento materno exclusivo, enquanto a depressão pós-parto configura-se como fator de risco.

RESUMEN Objetivo: evaluar la autoeficacia para amamantar, la presencia de síntomas de depresión en el período postparto y la asociación entre autoeficacia del amamantamiento y la depresión postparto, con la interrupción de la lactancia materna exclusiva. Método: se trata de un estudio de cohorte realizado entre 83 mujeres. Los instrumentos utilizados fueron la Escala de Autoeficacia en Lactancia Materna y la Escala de Depresión Postparto de Edimburgo, mediante análisis estadístico con las pruebas logrank, análisis de varianza y el modelo de supervivencia de Cox. Resultados: los niveles de autoeficacia para el amamantamiento (p=0,315) y depresión postparto (p=0,0879), a lo largo del tiempo, no demostraron diferencia estadística. Las oportunidades de interrupción de la lactancia materna exclusiva disminuyen el 48%, mejorando la autoeficacia de bajo a medio y el 80% de medio hacia alto. Puérperas con puntuación ≥10 en la Escala de Depresión Postparto de Edimburgo interrumpen la lactancia exclusiva, como promedido,10 días antes que aquellas con puntuación ≤9, cuya mediana de lactancia materna fue de 38 días después del parto. Conclusión: la autoeficacia para el amamantamiento se reveló como factor de protección para la lactancia materna exclusiva, mientras la depresión postparto se configura como un factor de riesgo.

Depressão pós-parto e autoeficácia materna para amamentar: prevalência e associação / Postpartum depression and maternal self-efficacy for breastfeeding: prevalence and association

Resumo Objetivo Identificar a prevalência de sintomas de depressão pós-parto e o nível de autoeficácia para amamentar, entre puérperas atendidas num Centro de Incentivo ao Aleitamento Materno, e analisar possíveis associações. Métodos Estudo transversal com 208 mulheres, até 60 dias pós-parto, submetidas à Escala de Depressão Pós-parto de Edinburgo e à Escala de Autoeficácia para Amamentar. Resultados Sintomas de depressão pós-parto estiveram presentes em 31,25% das mulheres, que apresentaram níveis de autoeficácia para amamentar médio (39,9%) e alto (36,06%). Ter média ou alta autoeficácia diminui em 27,4% ou 38,8%, respectivamente, o escore de depressão, enquanto a elevada pontuação na escala de depressão pós-parto reduz em 11,84 pontos o escore da autoeficácia na amamentação. Conclusão Prevalência elevada de sintomas de depressão pós-parto e de autoeficácia para amamentar foram evidenciados na população estudada. Os níveis de sintomas de depressão pós-parto e de autoeficácia revelaram associação de causa e efeito entre si.

Abstract Objective To identify the prevalence of postpartum depression symptoms and the level of self-efficacy to breastfeed among puerperal women assisted at a Breastfeeding Incentive Center, and to analyze possible associations. Methods A cross-sectional study with 208 women up to 60 days postpartum submitted to the Edinburgh Postpartum Depression Scale and the Self-Efficacy Scale for Breastfeeding. Results Postpartum depression symptoms were present in 31.25% of women who presented medium (39.9%) and high (36.06%) levels of breastfeeding self-efficacy. The medium or high self-efficacy decreased the depression score by 27.4% or 38.8% respectively, while the high score on the postpartum depression scale reduced the self-efficacy score in breastfeeding by 11.84 points. Conclusion The high prevalence of postpartum depression symptoms and self-efficacy for breastfeeding were evident in the studied population. The levels of postpartum depression symptoms and self-efficacy revealed a cause and effect association between themselves.

Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples

Abstract Background Several in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans. Methods A total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae. Results All used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains. Conclusions The in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.

Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples.

BACKGROUND: Several in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans. METHODS: A total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae. RESULTS: All used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains. CONCLUSIONS: The in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.

Haemophilus influenzae serotype b and a capsule-deficient type mutant (b-) invasive disease in a partially vaccinated child in Brazil.

We report a rare case of infection by two different types of Haemophilus influenzae strains in a child who received only one dose of the H. influenzae serotype b (Hib) conjugate vaccine (DTwP+Hib). The strains were recovered from blood and cerebrospinal fluid (CSF) and were phenotypically identified as Hib and non-typable H. influenzae, respectively, after serological tests. The two strains were characterized by PCR capsular typing, multilocus sequence typing and PFGE. Our results suggest that the infection was caused by the bloodstream invasion by a single Hib strain, followed by the diffusion of the bacteria across the blood-brain barrier and into the CSF. The strain recovered from the CSF, however, was identified as a capsule-deficient type mutant (b(-)) strain. Despite the high efficacy of the Hib conjugate vaccine, the increase in the numbers of strains able to escape the immune system of the vaccinated population advocates continued surveillance.

Changes in Haemophilus influenzae capsule locus: possible emergence of novel variants in Brazil.

A total of 28 strains of Haemophilus influenzae (Hi) a and b isolated from clinical samples before and after the introduction of the Hib conjugate vaccine in Brazil were analyzed to determine variants of the capsular gene. Our results suggest the occurrence of new variants closely related to types I and II previously described elsewhere. Eleven Hib strains belonged to type I, 8 were type II, and 3 Hia strains were type II. Six strains showed negative results after polymerase chain reaction targeting capsule locus; the variable regions were sequenced and compared with types I and II. Phylogenetic analysis showed that 5 Hib strains were actually subtypes of type I (type I-A), whereas 1 Hia strain was a subtype of type II (type II-A). Types I and II strains were present in both periods of vaccination. This study suggests that a gradual change in the capsule genes of H. influenzae is probably occurring, and novel variants might be emerging among Brazilian isolates.