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Nara Nóbrega Crispim Carvalho,1,2 Vinícius José Baccin Martins,3 João Modesto Filho,2 Francisco Bandeira,4 Flávia Cristina Fernandes Pimenta,5 José Luiz de Brito Alves1 1Department of Nutrition, Health Sciences Center, Federal University of Paraiba, Joao Pessoa, Brazil; 2Department of Endocrinology, Lauro Wanderley University Hospital, Federal University of Paraiba, Joao Pessoa, Brazil; 3Department of Physiology and Pathology, Federal University of Paraiba, Joao Pessoa, Brazil; 4Division of Diabetes and Endocrinology of Agamenon Magalhães Hospital, Recife, Brazil; 5Lauro Wanderley University Hospital, Federal University of Paraiba, Joao Pessoa, BrazilCorrespondence: José Luiz de Brito AlvesDepartment of Nutrition, Federal University of Paraiba, Campus I – Jd. Cidade Universitária, João Pessoa, PB 58051-900, BrazilTel/Fax +55 81 9 9845 5485Email jose_luiz_61@hotmail.com Nara Nóbrega Crispim CarvalhoDepartment of Endocrinology, Lauro Wanderley University Hospital, Federal University of Paraiba, João Pessoa, PB 58051-900, BrazilTel/Fax +55 83 9 9368 0937Email naranc@gmail.comBackground and aim: We evaluated low skeletal muscle mass (LMM) in women prior to bariatric surgery (BS) through different skeletal muscle mass indexes (MMIs) regarding body fat percentage (BFP), handgrip strength (HS), six-minute walk test (6MWT), metabolic profile and bone mineral density (BMD).Methods: Women (n=62) were allocated into two groups according to LMM: obesity with low muscle mass (OLMM) or obesity with normal muscle mass (ONMM). LMM was defined by the appendicular skeletal muscle mass (ASM) adjusted for weight (ASM/wt × 100) and ASM adjusted for body mass index (ASM/BMI), considering the lowest quintile of the indexes studied.Results: OLMM was found in 30.5% by ASM/wt × 100 and 20.3% by ASM/BMI. Using the ASM/wt × 100, OLMM group had a high BFP, low HS and BMD in L1-L4, femoral neck (FN) and total femur (TF) when compared with ONMM (p < 0.05). Using ASM/BMI, OLMM group had increased BFP, reduced HS and 6MWT in comparison to ONMM (p < 0.05). Metabolic profile was similar between OLMM and ONMM groups by the two MMIs. MMIs were negatively correlated with BFP (p < 0.05) and positively correlated with HS (p < 0.05), and none of them with 6MWT (p > 0.05). ASM/wt × 100 was positively correlated with all BMD sites assessed (p < 0.05). There was positive correlation between ASM/wt × 100 and ASM/BMI.Conclusion: OLMM identified by the ASM/wt × 100 and ASM/BMI had higher adiposity and lower HS. Using ASM/BMI, we found that OLMN had a poor physical performance, while the ASM/wt × 100 identified a lower BMD at all sites.Keywords: low muscle mass, obesity, muscle function, bone mass

Nara Nóbrega Crispim Carvalho,1,2 Francisco Antônio de Oliveira Junior,3 Gitana da Silva,4 Vinícius José Baccin Martins,3 Valdir de Andrade Braga,5 João Henrique da Costa-Silva,6 Flávia Cristina Fernandes Pimenta,4 José Luiz de Brito Alves11Department of Nutrition, Health Sciences Center, Federal University of Paraiba, João Pessoa, Brazil; 2Department of Endocrinology, Lauro Wanderley University Hospital, Federal University of Paraiba, João Pessoa, Brazil; 3Department of Physiology and Pathology, Health Sciences Center, Federal University of Paraiba, João Pessoa, Brazil; 4Lauro Wanderley Hospital, Federal University of Paraiba, João Pessoa, Brazil; 5Department of Biotechnology, Center of Biotechnology, Federal University of Paraíba, João Pessoa, Brazil; 6Department of Physical Education and Sport Sciences, Federal University of Pernambuco, Vitória De Santo Antão, PE, BrazilCorrespondence: José Luiz de Brito Alves; Nara Nóbrega Crispim CarvalhoDepartment of Nutrition, Federal University of Paraiba, Campus I – Jd. Cidade Universitária, João Pessoa, PB CEP: 58051-900, BrazilTel/fax +55 8 199 845 5485Email jose_luiz_61@hotmail.com; naranc@gmail.comBackground and aim: Obese individuals with recommendation for bariatric surgery (BS) exhibit increased cardiovascular risk. The association of obesity with comorbidities, such as arterial hypertension (HTN) and type 2 diabetes mellitus (T2DM) can worsen cardiovascular dysfunction. This study aimed to investigate the effect of HTN on cardiac autonomic function and whether diabetes exacerbates HTN-related impairment of autonomic function in obese subjects.Methods: Samples (n=63) were allocated to three groups: Obese without HTN and T2DM (n=29), Obese with HTN (OHTN, n=17) and OHTN with T2DM (OHTN+T2DM, n=17), in which anthropometric measures, body composition, blood biochemical parameters, blood pressure (BP) and heart rate variability (HRV) were evaluated.Results: The age was higher in OHTN+T2DM and OHTN than in obese groups (p

Copyright of Revista Eletrônica de Enfermagem is the property of Revista Eletronica de Enfermagem and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Eletrônica de Enfermagem is the property of Revista Eletronica de Enfermagem and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Eletrônica de Enfermagem is the property of Revista Eletronica de Enfermagem and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Eletrônica de Enfermagem is the property of Revista Eletronica de Enfermagem and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Eletrônica de Enfermagem is the property of Revista Eletronica de Enfermagem and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Revista Eletrônica de Enfermagem is the property of Revista Eletronica de Enfermagem and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Kenya introduced Synflorix™ (GlaxoSmithKline, PCV10-GSK), a 10-valent pneumococcal conjugate vaccine, in 2011, using three primary doses and, in select areas, catch-up campaigns. Surveys conducted 1-2 years post-introduction showed a stable prevalence of pneumococcal colonization, with declines in vaccine-type carriage. However, little is known about the long-term impact of PCV10-GSK in Kenya., Methods: We conducted a cross-sectional survey of pneumococcal carriage among children aged <5 years in November-December 2017 in Kibera (Nairobi informal settlement, no catch-up) and Asembo (rural western Kenya, 2-dose catch-up for children 1-4 years), using the same methods and settings as prior annual surveys from 2009 to 2013. Participants were randomly selected from an ongoing population-based surveillance platform. Nasopharyngeal swabs were frozen in skim milk-tryptone-glucose-glycerin media within 4 h and underwent culture with broth enrichment for pneumococcus. Isolates were serotyped by polymerase chain reaction and Quellung., Results: We enrolled 504 children, including 252 from each site; >90 % of participants had received 3 doses of PCV10-GSK. Pneumococcal colonization was detected in 210 (83.3 %) participants in Kibera and 149 (59.1 %) in Asembo, which was significantly lower than the prevalence observed in 2013 (92.9 % and 85.7 %, respectively). PCV10-GSK serotypes were detected in 35/252 (13.9 %) participants in Kibera and 23/252 (9.1 %) in Asembo, respectively; these prevalences were lower, but not statistically different, from vaccine-type carriage prevalences in 2013 (17.3 % and 13.3 %, respectively). In 2017 in both sites, serotypes 3, 6A, 19A, 19F, and 35B were among the most common serotypes., Conclusion: Six years post-PCV10-GSK introduction, the prevalence of pneumococcal carriage among children has decreased, and the impact of PCV10-GSK on vaccine-type carriage has plateaued. Kenya recently changed from PCV10-GSK to Pneumosil™ (Serum Institute of India), a 10-valent PCV that includes serotypes 6A and 19A; these data provide historical context for interpreting changes in vaccine-type carriage following the PCV formulation switch., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Ltd.)

Sugarcane juice is a nutritious and energetic drink. For its processing, the use of supercritical carbon dioxide (SC-CO 2 ) technology as an intervention potentially capable of rendering a high quality product can be considered. This study evaluated the combined effect of SC-CO 2 and mild temperatures, primarily aiming for the reduction of endogenous microorganisms and enzymes in sugarcane juice (pH~5.5). Pressures (P) ranging from 74 to 351 bar, temperatures (T) between 33 and 67 °C, and holding times (t) between 20 and 70 min were tested in a central composite rotational design. Seventeen trials were performed, comprising three replicates at the central points. Counts of aerobic mesophiles, molds and yeasts, lactic acid bacteria and coliforms at 45 °C, determination of polyphenol oxidase (PPO) and peroxidase (POD) activities, and measurement of color parameters in freshly extracted and processed juice's samples were carried out. The pH of fresh and processed juice varied between 4.6 and 6.0, and between 4.6 and 6.3, respectively. The number of decimal reductions achieved in mesophiles, molds and yeasts, lactic acid bacteria and coliforms varied between 0.1 and 3.9, 2.1 and 4.1, 0.0 and 2.1, and 0.3 to 2.5, respectively. The percentages of PPO reduction ranged from 3.51% to 64.18%. Regarding the POD, reductions between 0.27% and 41.42% were obtained. Color variations between fresh and processed samples varied between 2.0 and 12.3. As for mesophiles, molds and yeasts reduction, and soluble solids variation, none of the variables or their interactions were significant. In terms of polyphenol oxidase (PPO) reduction, only t was significant; however, T, t, and the interaction between them significantly affected the peroxidase (POD) reduction. In regards to pH variation, P, and the interaction between T and t were significant. P, T, t, and the interaction between T and t played a significant effect on color. The combination of mild temperatures and SC-CO 2 can be potentially used for cane juice preservation., (© 2024. The Author(s).)

Pneumococcal conjugate vaccines (PCVs) prevent nasopharyngeal colonization with vaccine serotypes of Streptococcus pneumoniae, leading to reduced transmission of pneumococci and stronger population-level impact of PCVs. In 2017 we conducted a cross-sectional pneumococcal carriage study in Indonesia among children aged <5 years before 13-valent PCV (PCV13) introduction. Nasopharyngeal swabs were collected during visits to community integrated health service posts at one peri-urban and one rural study site. Specimens were analyzed by culture, and isolates were serotyped using sequential multiplex polymerase chain and Quellung reaction. Antibiotic susceptibility was performed by broth microdilution method. We enrolled 1,007 children in Gunungkidul District, Yogyakarta (peri-urban) and 815 in Southwest Sumba, East Nusa Tenggara (rural). Pneumococcal carriage prevalence was 30.9% in Gunungkidul and 87.6% in Southwest Sumba (combined: 56.3%). PCV13 serotypes (VT) carriage was 15.0% in Gunungkidul and 52.6% in Southwest Sumba (combined: 31.8%). Among pneumococcal isolates identified, the most common VT were 6B (16.4%), 19F (15.8%), and 3 (4.6%) in Gunungkidul (N = 323) and 6B (17.6%), 19F (11.0%), and 23F (9.3%) in Southwest Sumba (N = 784). Factors associated with pneumococcal carriage were age (1-2 years adjusted odds ratio (aOR) 1.9, 95% CI 1.4-2.5; 3-4 years aOR 1.5, 95% CI 1.1-2.1; reference <1 year), other children <5 years old in the household (aOR 1.5, 95% CI 1.1-2.0), and presence of ≥1 respiratory illness symptom (aOR 1.8, 95% CI 1.4-2.2). Overall, 61.5% of the pneumococcal isolates were non-susceptible to ≥1 antibiotic class and 13.2% were multi-drug non-susceptible (MDNS) (non-susceptible to ≥3 classes of antibiotics). Among 602 VT isolates, 73.9% were non-susceptible and 19.9% were MDNS. These findings are critical to establish a pre-PCV13 carriage prevalence and demonstrate the complexity in evaluating the impact of PCV13 introduction in Indonesia given the wide variability in the carriage prevalence as shown by the two study sites., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Streptococcus pneumoniae is a cause of invasive diseases such as pneumonia, meningitis, and other serious infections among children and adults in Paraguay. This study was conducted to establish S. pneumoniae baseline prevalence, serotype distribution, and antibiotic resistance patterns in healthy children aged 2 to 59 months and adults ≥60 years of age prior to the introduction of PCV10 in the national childhood immunization program in Paraguay. Between April and July 2012, a total of 1444 nasopharyngeal swabs were collected, 718 from children aged 2 to 59 months and 726 from adults ≥60 years of age. The pneumococcal isolation, serotyping, and antibiotic susceptibility testing were performed using standard tests. Pneumococcal colonization prevalence was 34.1% (245/718) in children and 3.3% (24/726) in adults. The most frequent pneumococcal vaccine-types (VT) detected in the children were 6B (42/245), 19F (32/245), 14 (17/245), and 23F (20/245). Carriage prevalence with PCV10 serotypes was 50.6% (124/245) and PCV13 was 59.5% (146/245). Among colonized adults, prevalence of PCV10 and PCV13 serotypes were 29.1% (7/24) and 41.6% (10/24), respectively. Colonized children were more likely to share a bedroom, have a history of respiratory infection or pneumococcal infection compared to non-colonized children. no associations were found in adults. However, no significant associations were found in children and neither in adults. Vaccine-type pneumococcal colonization was highly prevalent in children and rare in adults in Paraguay prior to vaccine introduction, supporting the introduction of PCV10 in the country in 2012. These data will be useful to evaluate the impact of PCV introduction in the country., Competing Interests: The autors have declared that no competing interests exist., (Copyright: © 2023 Chamorro et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Minimally invasive tissue sampling (MITS) is an alternative to complete autopsy for determining causes of death. Multiplex molecular testing performed on MITS specimens poses challenges of interpretation, due to high sensitivity and indiscriminate detection of pathogenic, commensal, or contaminating microorganisms., Methods: MITS was performed on 20 deceased children with respiratory illness, at 10 timepoints up to 88 hours postmortem. Samples were evaluated by multiplex molecular testing on fresh tissues by TaqMan® Array Card (TAC) and by histopathology, special stains, immunohistochemistry (IHC), and molecular testing (PCR) on formalin-fixed, paraffin-embedded (FFPE) tissues. Results were correlated to determine overall pathologic and etiologic diagnoses and to guide interpretation of TAC results., Results: MITS specimens collected up to 3 days postmortem were adequate for histopathologic evaluation and testing. Seven different etiologic agents were detected by TAC in 10 cases. Three cases had etiologic agents detected by FFPE or other methods and not TAC; 2 were agents not present on TAC, and 2 were streptococci that may have been species other than those present on TAC. Result agreement was 43% for TAC and IHC or PCR, and 69% for IHC and PCR. Extraneous TAC results were common, especially when aspiration was present., Conclusions: TAC can be performed on MITS up to 3 days after death with refrigeration and provides a sensitive method for detection of pathogens but requires careful interpretation in the context of clinicoepidemiologic and histopathologic findings. Interpretation of all diagnostic tests in aggregate to establish overall case diagnoses maximizes the utility of TAC in MITS., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America.)

Group B Streptococcus (GBS) is a bacterial pathogen which is a leading cause of neonatal infection. Currently, there are limited GBS data available from the Indonesian population. In this study, GBS colonization, serotype distribution and antimicrobial susceptibility profile of isolates were investigated among pregnant women in Jakarta, Indonesia. Demographics data, clinical characteristics and vaginal swabs were collected from 177 pregnant women (mean aged: 28.7 years old) at 29-40 weeks of gestation. Bacterial culture identification tests and latex agglutination were performed for GBS. Serotyping was done by conventional multiplex PCR and antibiotic susceptibility testing by broth microdilution. GBS colonization was found in 53 (30%) pregnant women. Serotype II was the most common serotype (30%) followed by serotype III (23%), Ia and IV (13% each), VI (8%), Ib and V (6% each), and one non-typeable strain. All isolates were susceptible to vancomycin, penicillin, ampicillin, cefotaxime, daptomycin and linezolid. The majority of GBS were resistant to tetracycline (89%) followed by clindamycin (21%), erythromycin (19%), and levofloxacin (6%). The serotype III was more resistant to erythromycin, clindamycin, and levofloxacin and these isolates were more likely to be multidrug resistant (6 out of 10) compared to other serotypes. This report provides demographics of GBS colonization and isolate characterization in pregnant women in Indonesia. The results may facilitate preventive strategies to reduce neonatal GBS infection and improve its treatment., Competing Interests: The authors have declared that no competing interests exist.

The polysaccharide capsule is a key virulence factor of Streptococcus pneumoniae There are numerous epidemiologically important pneumococcal capsular serotypes, and recent findings have demonstrated that several of them are commonly found among nonpathogenic commensal species. Here, we describe 9 nonpneumococcal strains carrying close homologs of pneumococcal capsular biosynthetic ( cps ) loci that were discovered during recent pneumococcal carriage studies of adults in the United States and Kenya. Two distinct Streptococcus infantis strains cross-reactive with pneumococcal serotype 4 and carrying cps4 -like capsular biosynthetic ( cps ) loci were recovered. Opsonophagocytic killing assays employing rabbit antisera raised against S. infantis US67cps4 revealed serotype 4-specific killing of both pneumococcal and nonpneumococcal strains. An S. infantis strain and two Streptococcus oralis strains, all carrying cps9A- like loci, were cross-reactive with pneumococcal serogroup 9 strains in immunodiffusion assays. Antiserum raised against S. infantis US64cps9A specifically promoted killing of serotype 9A and 9V pneumococcal strains as well as S. oralis serotype 9A strains. Serotype-specific PCR of oropharyngeal specimens from a recent adult carriage study in the United States indicated that such nonpneumococcal strains were much more common in this population than serotype 4 and serogroup 9 pneumococci. We also describe S. oralis and S. infantis strains expressing serotypes identical or highly related to serotypes 2, 13, and 23A. This study has expanded the known overlap of pneumococcal capsular serotypes with related commensal species. The frequent occurrence of nonpneumococcal strains in the upper respiratory tract that share vaccine and nonvaccine capsular serotypes with pneumococci could affect population immunity to circulating pneumococcal strains. IMPORTANCE The distributions and frequencies of individual pneumococcal capsular serotypes among nonpneumococcal strains in the upper respiratory tract are unknown and potentially affect pneumococcal serotype distributions among the population and immunity to circulating pneumococcal strains. Repeated demonstration that these nonpneumococcal strains expressing so-called pneumococcal serotypes are readily recovered from current carriage specimens is likely to be relevant to pneumococcal epidemiology, niche biology, and even to potential strategies of employing commensal live vaccines. Here, we describe multiple distinct nonpneumococcal counterparts for each of the pneumococcal conjugate vaccine (PCV) serotypes 4 and 9V. Additional data from contemporary commensal isolates expressing serotypes 2, 13, and 23A further demonstrate the ubiquity of such strains. Increased focus upon this serological overlap between S. pneumoniae and its close relatives may eventually prove that most, or possibly all, pneumococcal serotypes have counterparts expressed by the common upper respiratory tract commensal species Streptococcus mitis , Streptococcus oralis , and Streptococcus infantis ., (Copyright © 2021 Gertz et al.)

Background: Mozambique introduced 10-valent pneumococcal conjugate vaccine (PCV10) in 2013 with doses at ages 2, 3, and 4 months and no catch-up or booster dose. We evaluated PCV10 impact on the carriage of vaccine-type (VT), non-VT, and antimicrobial non-susceptible pneumococci 3 years after introduction., Methods: We conducted cross-sectional carriage surveys among HIV-infected and HIV-uninfected children aged 6 weeks to 59 months: 1 pre-PCV10 (2012-2013 [Baseline]) and 2 post-PCV10 introductions (2014-2015 [Post1] and 2015-2016 [Post2]). Pneumococci isolated from nasopharyngeal swabs underwent Quellung serotyping and antimicrobial susceptibility testing. Non-susceptible isolates (intermediate or resistant) were defined using Clinical and Laboratory Standards Institute 2018 breakpoints. We used log-binomial regression to estimate changes in the pneumococcal carriage between survey periods. We compared proportions of non-susceptible pneumococci between Baseline and Post2., Results: We enrolled 720 children at Baseline, 911 at Post1, and 1208 at Post2. Baseline VT carriage was similar for HIV-uninfected (36.0%, 110/306) and HIV-infected children (34.8%, 144/414). VT carriage was 36% (95% confidence interval [CI]: 19%-49%) and 27% (95% CI: 11%-41%) lower in Post1 vs baseline among HIV-uninfected and HIV-infected children, respectively. VT carriage prevalence declined in Post2 vs Post1 for HIV-uninfected but remained stable for HIV-infected children. VT carriage prevalence 3 years after PCV10 introduction was 14.5% in HIV-uninfected and 21.0% in HIV-infected children. Pneumococcal isolates non-susceptible to penicillin declined from 66.0% to 56.2% (P= .0281) among HIV-infected children., Conclusions: VT and antimicrobial non-susceptible pneumococci carriage dropped after PCV10 introduction, especially in HIV-uninfected children. However, VT carriage remained common, indicating ongoing VT pneumococci transmission., (Published by Oxford University Press on behalf of The Journal of the Pediatric Infectious Diseases Society 2020.)

We compared pneumococcal isolation rates and evaluated the benefit of using oropharyngeal (OP) specimens in addition to nasopharyngeal (NP) specimens collected from adults in rural Kenya. Of 846 adults, 52.1% were colonized; pneumococci were detected from both NP and OP specimens in 23.5%, NP only in 22.9%, and OP only in 5.7%. Ten-valent pneumococcal conjugate vaccine strains were detected from both NP and OP in 3.4%, NP only in 4.1%, and OP only in 0.7%. Inclusion of OP swabs increased carriage detection by 5.7%; however, the added cost of collecting and processing OP specimens may justify exclusion from future carriage studies among adults., (© The Author(s) 2020. Published by Oxford University Press on behalf of Infectious Diseases Society of America.)

Background: Dried blood spots (DBS) have been proposed as potentially tool for detecting invasive bacterial diseases., Methods: We evaluated the use of DBS for S. pneumoniae and H. influenzae detection among children in Mozambique. Blood for DBS and nasopharyngeal (NP) swabs were collected from children with pneumonia and healthy aged < 5 years. Bacterial detection and serotyping were performed by quantitative PCR (qPCR) (NP and DBS; lytA gene for pneumococcus and hpd for H. influenzae) and culture (NP). Combined detection rates were compared between children with pneumonia and healthy., Results: Of 325 children enrolled, 205 had pneumonia and 120 were healthy. Pneumococci were detected in DBS from 20.5 and 64.2% of children with pneumonia and healthy, respectively; NP specimens were positive for pneumococcus in 80.0 and 80.8%, respectively. H. influenzae was detected in DBS from 22.9% of children with pneumonia and 59.2% of healthy; 81.4 and 81.5% of NP specimens were positive for H. influenzae, respectively., Conclusion: DBS detected pneumococcal and H. influenzae DNA in children with pneumonia and healthy. Healthy children were often DBS positive for both bacteria, suggesting that qPCR of DBS specimens does not differentiate disease from colonization and is therefore not a useful diagnostic tool for children.

Purpose: Physical exercise is associated with reduced blood pressure (BP). Moderate-intensity continuous exercise (MCE) promotes post-exercise hypotension (PEH), which is highly recommended to hypertensive patients. However, recent studies with high-intensity interval exercise (HIIE) have shown significant results in cardiovascular disease. Thus, this study aimed to analyze PEH in hypertensive subjects submitted to HIIE and compare it to post MCE hypotension., Methods: 20 hypertensive adults (51 ± 8 years), treated with antihypertensive medications, were submitted to two different exercise protocols and a control session. The MCE was performed at 60-70% of VO 2 reserve, while HIIE was composed of five bouts of 3 min at 85-95% VO 2 reserve with 2 min at 50% of VO 2 reserve. The following variables were evaluated during exercise, pre- and post-session: clinical BP, heart rate (HR), double product, perception of effort, body mass, height and body mass index., Results: Systolic BP decreased after exercise in both sessions, showing greater decrease after HIIE (- 7 ± 10 and - 11 ± 12 mmHg, after MCE and HIIE, respectively, p ≤ 0.01). Diastolic BP also decreased after both sessions, but there were no significant differences between the two sessions (- 4 ± 8 and - 7 ± 8 mmHg, after MCE and HIIE, respectively)., Conclusion: Both exercise sessions produced PEH, but HIIE generated a greater magnitude of hypotension. The HIIE protocol performed in this study caused a greater cardiovascular stress during exercise; however, it was safe for the studied population and efficient for reducing BP after exercise.

The bone marrow aspiration procedure is used in hematological diseases and consists of a painful, invasive procedure causing anxiety-associated symptoms. The present study assessed the effect of Citrus aurantium L. essential oil on the treatment of anxiety, in the moment that precedes the collection of medullary material in patients with chronic myeloid leukemia (CML). Volunteers from both sexes were divided into groups receiving either the C. aurantium essential oil through inhalation, diazepam (10 mg), or the placebo. The evaluation was performed through psychometric scales [State-Trait Anxiety Inventory (STAI)] and physiological measurements (blood pressure and cardiac and respiratory frequency). Inhalation of C. aurantium was associated with a decrease in the STAI-S scores, suggesting an anxiolytic effect. In support of these results, a change in all the physiological measurements was observed in the group exposed to C. aurantium. In the diazepam group, only the diastolic pressure decreased, and no effect was observed in the placebo group. Therefore, the results showed that C. aurantium exhibits an anxiolytic effect and reduces the signs and symptoms associated with anxiety in patients with CML., (Copyright © 2016 John Wiley & Sons, Ltd.)

Polymerase chain reaction-based surveillance for bacterial meningitis including 841 children revealed 246 with bacterial DNA in cerebrospinal fluid samples of which 53% were Streptococcus pneumoniae, 19% Neisseria meningitidis, and 16% Haemophilus influenzae type b. The most common S. pneumoniae serotypes/serogroups were 1, 19F, 6A/6B, 23F, 5, 14, 18 and 19A. Among 47 meningococci, 86% were serogroup B, 6% serogroup C, 3% serogroup A, 3% serogroup X and 3% serogroup W.

Background: 10-valent conjugate pneumococcal vaccine/PCV10 was introduced in the Brazilian National Immunization Program along the year of 2010. We assessed the direct effectiveness of PCV10 vaccination in preventing nasopharyngeal/NP pneumococcal carriage in infants., Methods: A cross-sectional population-based household survey was conducted in Goiania Brazil, from December/2010-February/2011 targeting children aged 7-11 m and 15-18 m. Participants were selected using a systematic sampling. NP swabs, demographic data, and vaccination status were collected from 1,287 children during home visits. Main outcome and exposure of interest were PCV10 vaccine-type carriage and dosing schedules (3p+0, 2p+0, and one catch-up dose), respectively. Pneumococcal carriage was defined by a positive culture and serotyping was performed by Quellung reaction. Rate ratio/RR was calculated as the ratio between the prevalence of vaccine-types carriage in children exposed to different schedules and unvaccinated for PCV10. Adjusted RR was estimated using Poisson regression. PCV10 effectiveness/VE on vaccine-type carriage was calculated as 1-RR*100., Results: The prevalence of pneumococcal carriage was 41.0% (95%CI: 38.4-43.7). Serotypes covered by PCV10 and PCV13 were 35.2% and 53.0%, respectively. Vaccine serotypes 6B (11.6%), 23F (7.8%), 14 (6.8%), and 19F (6.6%) were the most frequently observed. After adjusted for confounders, children who had received 2p+0 or 3p+0 dosing schedule presented a significant reduction in pneumococcal vaccine-type carriage, with PCV10 VE equal to 35.9% (95%CI: 4.2-57.1; p = 0.030) and 44.0% (95%CI: 14.-63.5; p = 0.008), respectively, when compared with unvaccinated children. For children who received one catch-up dose, no significant VE was detected (p = 0.905)., Conclusion: PCV10 was associated with high protection against vaccine-type carriage with 2p+0 and 3p+0 doses for children vaccinated before the second semester of life. The continuous evaluation of carriage serotypes distribution is likely to be useful for evaluating the long-term effectiveness and impact of pneumococcal vaccination on serotypes reduction.

We performed culture-based and PCR-based tests for pneumococcal identification and serotyping from carriage specimens collected in rural and urban Kenya. Nasopharyngeal specimens from 237 healthy children <5 years old (C-NPs) and combined nasopharyngeal/oropharyngeal specimens from 158 adults (A-NP/OPs, 118 HIV-positive) were assessed using pneumococcal isolation (following broth culture enrichment) with Quellung-based serotyping, real-time lytA-PCR, and conventional multiplexed PCR-serotyping (cmPCR). Culture-based testing from C-NPs, HIV-positive A-NP/OPs, and HIV-negative A-NP/OPs revealed 85.2%, 40.7%, and 12.5% pneumococcal carriage, respectively. In contrast, cmPCR serotypes were found in 93.2%, 98.3%, and 95.0% of these sets, respectively. Two of 16 lytA-negative C-NPs and 26 of 28 lytA-negative A-NP/OPs were cmPCR-positive for 1-10 serotypes (sts) or serogroups (sgs). A-NP/OPs averaged 5.5 cmPCR serotypes/serogroups (5.2 in HIV-positive, 7.1 in HIV-negative) and C-NPs averaged 1.5 cmPCR serotypes/serogroups. cmPCR serotypes/serogroups from lytA-negative A-NP/OPs included st2, st4, sg7F/7A, sg9N/9L, st10A, sg10F/10C/33C, st13, st17F, sg18C/18A/18B/18F, sg22F/22A, and st39. Nine strains of three non-pneumococcal species (S. oralis, S. mitis, and S. parasanguinis) (7 from A-OP, 1 from both A-NP and A-OP, and 1 from C-NP) were each cmPCR-positive for one of 7 serotypes/serogroups (st5, st13, sg15A/15F, sg10F/10C/33C, sg33F/33A/37, sg18C/18A/18B/18F, sg12F/12A/12B/ 44/46) with amplicons revealing 83.6-99.7% sequence identity to pneumococcal references. In total, 150 cmPCR amplicons from carriage specimens were sequenced, including 25 from lytA-negative specimens. Amplicon sequences derived from specimens yielding a pneumococcal isolate with the corresponding serotype were identical or highly conserved (>98.7%) with the reference cmPCR amplicon for the st, while cmPCR amplicons from lytA-negative specimens were generally more divergent. Separate testing of 56 A-OPs and 56 A-NPs revealed that ∼94% of the positive cmPCR results from A-NP/OPs were from OP microbiota. In contrast, A-NPs yielded >2-fold more pneumococcal isolates than A-OPs. Verified and suspected non-pneumococcal cmPCR serotypes/serogroups appeared to be relatively rare in C-NPs and A-NPs compared to A-OPs. Our findings indicate that non-pneumococcal species can confound serotype-specific PCR and other sequence-based assays due to evolutionarily conserved genes most likely involved in biosynthesis of surface polysaccharide structures.

We developed and validated a real-time PCR assay consisting of 7 triplexed reactions to identify 11 individual serotypes plus 10 small serogroups representing the majority of disease-causing isolates of Streptococcus pneumoniae. This assay targets the 13 serotypes included within the 13-valent conjugate vaccine and 8 additional key serotypes or serogroups. Advantages over other serotyping assays are described. The assay will be expanded to 40 serotypes/serogroups. We will provide periodic updates at our protocol website.

The traditional use of essential oils in aromatherapy has offered numerous health benefits. However, few scientific studies have been conducted with these oils to confirm their therapeutic efficacy. (+)-Limonene is a chemical constituent of various bioactive essential oils. The present study reports on the anxiolytic-like effects of (+)-limonene in an elevated maze model of anxiety in mice. At concentrations of 0.5% and 1.0%, (+)-limonene, administered to mice by inhalation, significantly modified all the parameters evaluated in the elevated plus maze test. The pharmacological effect of inhaled (+)-limonene (1%) was not blocked by flumazenil. Analysis of (+)-limonene using gas chromatography-mass spectrometry (GC-MS) showed its volatility to be high. These data suggest possible connections between the volatility of (+)-limonene and its anxiolytic-like effect on the parameters evaluated in the elevated plus maze test. The data indicate that (+)-limonene could be used in aromatherapy as an antianxiety agent., (Copyright © 2012 Elsevier Inc. All rights reserved.)

A child's death due to pneumococcal meningitis after contracting the disease in an after-school programme prompted an investigation to assess nasopharyngeal (NP) carriage among her contacts. The serotype of the meningitis case isolate was determined, together with the serotypes of the NP specimens of contacts, comprising the case patient's brother, the case patient's after-school programme contacts and the brother's day-care centre (DCC) contacts. NP swabs from 155 children and 69 adults were obtained. Real-time PCR and conventional multiplex PCR (CM-PCR) assays were used to detect pneumococcal carriage and determine serotypes. Broth-enriched culture of NP specimens followed by pneumococcal isolation and Quellung-based serotyping were also performed. DNA extracts prepared from cerebrospinal fluid of the index case and from the NP strain isolated from the brother and from one attendee of the brother's DCC were subjected to genotyping. Pneumococcal carriage assessed by real-time PCR and culture was 49.6 and 36.6%, respectively (P<0.05). Twenty-three serotypes were detected using CM-PCR, with serotypes 6A/6B, 14, 19F, 6C/6D, 22F/22A, 23F and 11A/11D being the most frequent. All eight serotype 22F/22A NP specimens recovered were from children attending the brother's DCC. The meningitis case isolate and the NP carriage isolate from the patient's brother were both serotype 22F and shared the same new multilocus sequence type (ST6403) with the attendee of the brother's DCC. CM-PCR proved to be useful for assessing carriage serotype distribution in a setting of high-risk pneumococcal transmission. The causal serotype appeared to be linked to the brother of the case patient and attendees of his DCC., (© 2012 SGM)

This study investigated the presence of the black-pigmented bacteria Prevotella nigrescens and Prevotella intermedia, the non-black-pigmented bacteria Actinomyces spp and particularly the cariogenic pathogen Streptococcus mutans in the dental biofilms of patients with or without black extrinsic tooth stains, using the multiplex polymerase chain reaction (PCR) technique. Analysis of the dental biofilms of patients with (n=26) or without (n=26) black tooth stains was performed using duplex PCR for the 16S ribosomal RNA gene (P. nigrescens, P. intermedia, Actinomyces spp) and glucosyltransferase-I gene for S. mutans. P. nigrescens and S. mutans were the most frequent bacteria detected in both groups. The least frequently detected were P. intermedia and Actinomyces spp. The similar bacterial composition of dental biofilms of black tooth stains and healthy tooth surfaces indicates that black tooth stains are not free of cariogenic bacteria.

The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Norway is low, compared to other European and American countries. The health system includes mandatory case reporting and has written guidelines for prevention and control. This communication describes the national public policies related to MRSA obtained from documents and academic experience gained during a doctoral fellowship in Oslo, Norway. The painstaking procedures used for investigating suspected cases, including health professionals, decolonization and case monitoring, could be important tools to be used by countries with a high prevalence of MRSA.

Pneumococcal nasopharyngeal carriage isolates recovered from Brazilian children attending day-care centres in 2005 were assessed for serotype, genotype and penicillin susceptibility phenotype. As 124 of the 253 isolates (49 %) were characterized previously with respect to serotype and penicillin susceptibility, the primary objectives were to examine clonal associations and penicillin susceptibility within major serotypes and to assess the suitability of conventional multiplex PCR for deducing carriage serotypes within this population. Using a combination of PCR-based serotyping and the Quellung reaction, serotypes were identified for 81 % (205/253) of the isolates, with serogroups or types 14, 6, 23F, 19F and 18 being predominant. Included within the 205 isolates successfully serotyped by PCR were 28 isolates that had become non-viable. Forty-eight isolates were non-typable using both the PCR method and the Quellung reaction. Penicillin non-susceptibility was observed within 16 of the 18 multilocus sequence types detected. Thus, this study provides further evidence from a diverse collection of pneumococcal clones that PCR-based serotype deduction is useful for providing supportive evidence for pneumococcal conjugate vaccine implementation.

Introduction: The increasing prevalence of enterococci strains in hospitals, particularly among isolates of vancomycin-resistant enterococci (VRE), poses important problems because of the limited effect of antimicrobial therapy for enterococcal infections., Methods: This work presents a retrospective investigation of quantitative in vitro susceptibility data for the range of antimicrobials against Enterococcus spp. isolates and evaluation of the association of resistance between antimicrobial agents recommended as the treatment of choice for infections caused by VRE through calculation of the relative risk., Results: Of the 156 enterococci isolates, 40 (25.6%) were resistant to 3 or more antimicrobials, including 7.7% (n = 12/156) vancomycin resistant. The association of elevated resistance was more pronounced among VRE isolates against alternative and primary antimicrobials for the treatment of infections caused by these pathogens, including ampicillin (100%, RR = 7.2), streptomycin (90.9%, RR = 4.9), rifampin (91.7%, RR = 3.1) and linezolid (50%, RR = 11.5), despite high susceptibility to this drug (94.9%)., Conclusions: The significant associated resistance to alternative and first choice antimicrobials used in the treatment of serious infections of strains with the VRE phenotype and that require a combined therapeutic regime, revealed even more limited therapeutic alternatives in the institution analyzed.

This cross-sectional study aimed to investigate the presence of Staphylococcus aureus in the saliva of the nursing team of a teaching hospital in the interior of São Paulo State. Three saliva samples were collected from 351 individuals with an interval of two months between each collection. All ethical aspects were considered. In 867 (82.3%) cultures there was no identification of Staphylococcus aureus in the saliva, in 88 (17.7%) cultures Staphylococcus aureus was isolated, 26 (2.5%) of which were resistant to methicillin. The prevalence of professionals colonized by Staphylococcus aureus was 41.0% (144/351), of which 7.1% (25/351) were characterized as methicillin-resistant Staphylococcus aureus. Transient carriers represented 81.2% and persistent carriers 18.8%. Resistance to mupirocin was 73.1% of MRSA and 9.3% of MSSA. The results demonstrate that it is the nurse and nursing technician that are the professional categories most susceptible to MRSA. Broader discussion on the thematic and interventions are needed.

PspA is one of the most well studied pneumococcal proteins and a promising candidate for a future protein-based anti-pneumococcal vaccine. Nevertheless, its structural and serological variability suggests the inclusion of more than one PspA molecule in order to broaden protection. Since different PspAs exhibit variable levels of cross-reactivity, the selection of the protein combination with the highest coverage potential is an essential step for PspA-based vaccine development. This work investigated the level of cross-reactivity within family 1 PspAs, and established a complement based antibody mediated opsonophagocytic assay for measuring the level of cross-protection. Among a panel of ten family 1 PspA molecules, two of them, one belonging to clade 1 and another from clade 2, induced antibodies capable of enhancing complement deposition and mediating the phagocytic killing by mouse peritoneal macrophages of all pneumococci bearing PspA family 1 strains tested, regardless of their serotype. Therefore, we suggest the inclusion of either one in a PspA-based vaccine, as a representative of family 1. Furthermore, our results suggest that opsonophagocytosis by mouse peritoneal cells can be an efficient means of evaluating the induction of protective immune responses in mice across a large number of strains., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The aims of this study were to evaluate the incidence of mutans streptococci (MS - sessile form) on complete maxillary dentures after use of a specific denture paste, and to determine the minimum inhibitory concentration (MIC) and maximum inhibitory dilution (MID) of 3 oral mouthrinses: Cepacol, Plax and Periogard. Seventy-seven complete denture wearers were randomly assigned into 2 groups, according to the product used for denture cleaning: Control group - conventional dentifrice (Kolynos-Super White); and Test group: experimental denture cleaning paste. Denture biofilm was collected at baseline and after 90 and 180 days after treatment by brushing the dentures with saline solution. After decimal serial dilution, samples were seeded onto agar sucrose bacitracin to count colonies with morphological characteristics of MS. MS identification was performed by the sugar fermentation tests. After this procedure, brain heart infusion broth (BHI) was added to oral mouthrinses (Plax, Cepacol e Periogard) and seeded on Petri dishes. The colonies were seeded using the Steers multiplier and, after the incubation, the MIC and MID of the mouthrinses were calculated. The results showed an incidence of 74.0% (n=57) of MS in the 77 complete dentures examined in the study, being 76.3% (n=29) of the Control group (conventional dentifrice) and 71.8% (28) of the Test group (experimental denture cleaning paste). In both groups, the number of positive cases for MS decreased from day 0 to day 180. In the Test group there was a slight decrease in the incidence of Streptococcus mutans 90 days after use of the experimental denture cleaning paste, which was not observed in the Control group. As regards to mouthrinses, for both groups, Periogard showed antimicrobial action with the highest dilution, followed by Cepacol and Plax. In conclusion, the incidence of MS in complete dentures was high and Periogard was the mouthrinse with the strongest antimicrobial action against MS. The experimental denture cleaning paste showed a slight action against S. mutans after 90 days of treatment.

This study assessed the prevalence of colonization by Staphylococcus aureus in hospital housekeepers, and their knowledge and beliefs regarding this problem. Three saliva samples were collected and a questionnaire regarding knowledge and beliefs was applied. Of the 92 workers, 63 (68.5%) participated in the study; 20 were not and 43 were colonized; 13 by methicillin resistant Staphylococcus aureus and 30 by methicillin sensitive Staphylococcus aureus. Persistent carrier status of methicillin resistant Staphylococcus aureus was detected in 15.4% of cases. Low knowledge and perception of occupational risk were observed. The mouth was identified as an important reservoir of methicillin resistant Staphylococcus aureus. Analyzing knowledge and beliefs, as well as the state of carrier, is an important strategy to be added to educational actions for the prevention of workers' colonization.

Background: A second-generation 13-valent pneumococcal conjugate vaccine, PCV13, was recently licensed. Although PCV13 includes serotype 6A, the usefulness of that antigen may be limited by the emergence of a new serotype, 6C, which was identified among isolates initially characterized (Quellung reaction) as serotype 6A. The epidemiology of serotype 6C prior to and after 7-valent PCV (PCV7) introduction is incompletely understood., Methods: We analyzed conventionally serotyped 6A (CS6A) pneumococci from invasive disease case patients of all ages and carriage isolates from children and adults obtained in population-based studies among Navajo and White Mountain Apache communities during 1994-2009. Samples were tested by triplex polymerase chain reaction to resolve serotypes 6C and 6A., Results: A total of 74 invasive CS6A episodes occurred. All were retyped by polymerase chain reaction; 40 (54.1%) were serotype 6C. The mean annual incidence of serotype 6C invasive disease was 0.3 (95% confidence interval, 0.03-0.9), 0.7 (95% confidence interval, 0.2-1.3), and 1.5 (95% confidence interval, 1.0-2.1) cases per 100,000 population in the years prior to the PCV7 efficacy trial, during the time the PCV7 trial was conducted, and following PCV7 introduction and routine use, respectively (P = .01). In the routine vaccination era, 76% of invasive CS6As were serotype 6C; nearly all cases occurred in adults. The proportion of serotype 6C among CS6A carriage isolates increased from 42% to 61% to 94% in the prevaccine, early vaccine, and routine vaccination eras, respectively., Conclusion: In the PCV7 routine use era, virtually all serogroup 6 invasive pneumococcal disease and carriage strains among Navajo and White Mountain Apache communities are 6C. Monitoring and evaluation of this and other emerging serotypes among invasive disease and carriage isolates is warranted.

Background: During previous influenza pandemics, many deaths were associated with secondary bacterial infection. In April 2009, a previously unknown 2009 influenza A virus (2009 H1N1) emerged, causing a global influenza pandemic. We examined the relationship between circulating 2009 H1N1 and the occurrence of secondary bacterial parapneumonic empyema in children., Methods: Children hospitalized with parapneumonic empyema from August 2004 to July 2009, including a period when the 2009 H1N1 circulated in Utah, were identified using International Classification of Diseases, Ninth Revision codes. We compared the average number of children diagnosed with influenza A and the number of admissions for empyema per month for the previous 4 seasons to rates of empyema during the 2009 H1N1 outbreak. We identified causative bacteria using culture and polymerase chain reaction (PCR)., Results: We observed an increase in hospitalization of children with pneumonia complicated by empyema during a severe outbreak of 2009 H1N1 during the spring and summer of 2009, compared with historical data for the previous 4 seasons. Streptococcus pneumoniae and Streptococcus pyogenes were the predominant bacteria identified., Conclusions: Similar to previous pandemics, secondary bacterial infection with S. pneumoniae and S. pyogenes were associated with the 2009 H1N1 outbreak. There is an urgent need to better understand bacterial complications of pandemic influenza. In the interim, influenza vaccines, antiviral agents, and pneumococcal vaccines should be used to prevent cases of secondary bacterial pneumonia whenever possible.

Context: The endoscopic procedure safety depends on the use of an adequately reprocessed device which quality is related to each of its operational steps., Objective: To characterize the reprocessing of endoscopes using glutaraldehyde in endoscopy services, Methods: Study was conducted by observing the reprocessing of 60 endoscopes from 20 medical practices of the municipality of Goiânia, GO, central area of Brazil., Results: This study showed failure in all reprocessing steps. The pre-washing was performed in 24 (40.0%) of the endoscope. In the cleaning steps, was identify the improper use of enzymatic detergent, and in 27 (45.0%) cases, the brushing of internal channels was not performed. All 60 endoscopes were submitted to this disinfectant. However, for 33 (55.0%) of the cases the internal channels was not filled. The total immersion of endoscope in the glutaraldehyde was not performed in 39 (65.0%) cases. The recommended minimum total immersion time for exposure to 2% glutaraldehyde solution was followed only for 12 (20.0%) endoscopes. There was no filter for water treatment used in the rinse of most endoscopes 54 (90.0%) and to dry the internal channels only 6 (10.0%) of them used compressed air. Adequate storing conditions were identified., Conclusion: Considering the particularities of the endoscope and its reprocessing, it is imperative to establish protocols to ensure the quality of the disinfection and the prevention of cross-contamination.

Objective: The aim of this study was to detect the prevalence of selected bacterial species in intraoral sites of patients with chronic periodontitis (CP) using multiplex polymerase chain reaction (PCR)., Methodology: Samples were collected from the tongue dorsum, buccal mucosa, supragingival and subgingival plaque and saliva of 30 patients with untreated CP. Multiplex PCR was used to determine prevalence rates, which were then compared using a chi-square test. Significance level was set at p<0.05. Mean and standard deviation values were used to evaluate variations in prevalence according to site., Results: The prevalence of S. mutans was 70% in saliva; 60% in samples collected from the tongue dorsum; 50% in samples collected from the buccal mucosa; 56.5% in the supragingival plaque; and 53.5% in the subgingival plaque. The prevalence of E. faecalis ranged from 3.5% to 13.5% in all intraoral microenvironment. The highest prevalence of P. gingivalis was found in subgingival plaque (53.5%), and of P. intermedia in supragingival plaque (33.5%), subgingival plaque (30%) and tongue dorsum (33.5%). The prevalence of bacteria did not vary significantly among the intraoral sites., Conclusions: All studied bacteria were identified in intraoral sites. S. mutans, P. gingivalis and P. intermedia had high prevalence rates, but the prevalence of E. faecalis was low. Multiplex PCR proved to be an adequate method for epidemiological studies.

The measurement of pneumococcal carriage in the nasopharyngeal reservoir is subject to potential confounders that include low-density and multiple-strain colonization. To compare different methodologies, we picked a random sampling of 100 nasopharyngeal specimens recovered from infants less than 2 years of age who were previously assessed for pneumococcal carriage and serotypes by a conventional method that used direct plating from the transport/storage medium (50 specimens were culture negative and 50 specimens were culture positive for pneumococci). We used a broth enrichment approach and a conventional PCR approach (with and without broth enrichment) to determine pneumococcal carriage and serotypes, and the results were compared to the initial conventional culture-based results. Additionally, we used a lytA-targeted real-time PCR for pneumococcal detection. Broth enrichment for both the culture-based and the PCR-based methods enhanced the isolation of pneumococci and detection of serotype diversity, with the most effective serotype deduction method being one that used broth enrichment prior to sequential multiplex PCR. Similarly, we also found that broth enrichment followed by the lytA-specific real-time PCR was the most sensitive for the detection of apparent pneumococcal carriage. The broth enrichment, conventional multiplex PCR, and real-time PCR approaches used in this study were effective in detecting pneumococcal carriage in the 50 specimens that were negative by conventional direct plating from transport medium (range of numbers of positive specimens, 8/50 to 22/50 [16 to 44%]), and the three different serotyping approaches that used broth enrichment increased the number of serotype identifications from the 100 specimens (12 to 29 additional serotype identifications to be positive). A PCR-based approach that employed a broth enrichment step appeared to best enhance the detection of mixed serotypes and low-density pneumococcal carriage.

According to population-based invasive pneumococcal surveillance in the United States during 2007, 898 (26%) of 3,511 isolates were penicillin nonsusceptible. Non-7-valent pneumococcal conjugate vaccine (PCV7) serotypes other than 19A accounted for 40% of these penicillin-nonsusceptible isolates; of these, serotypes 15A (11%), 23A (8%), 35B (8%), and 6C (5%) were most common (cumulatively 32% of penicillin-nonsusceptible isolates). Each except 6C represented a single serotype and clonal complex combination that predated the introduction of PCV7. We evaluated the genetic characteristics and nonsusceptibility to penicillin of non- PCV7 serotypes, and we found increased proportions of specific penicillin-nonsusceptible clones in serotypes 15A, 23A, 35B, and 6C, which potentially indicates a basic change of population structure within these individual serotypes.

PspA is an important candidate for a vaccine with serotype-independent immunity against pneumococcal infections. Based on sequence relatedness, PspA has been classified into three families comprising six clades. We have previously addressed the cross-reactivity of antibodies against PspA fragments containing the N-terminal and proline-rich regions of PspA from clades 1 to 5 (PspA1, PspA2, PspA3, PspA4, and PspA5) by Western blot analysis and reported that anti-PspA4 and anti-PspA5 were able to recognize pneumococci expressing PspA proteins from all of the clades analyzed. We have now analyzed the functional capacity of these antibodies to bind and to mediate complement deposition on intact bacteria in vitro. Our results show that both PspA4 and PspA5 elicit antibodies that are able to bind and to mediate complement deposition efficiently on pneumococcal strains bearing PspA proteins from clades 1 to 5. Moreover, mice immunized with PspA4 and PspA5 were protected against an intranasal lethal challenge with strains expressing PspA proteins from the two major families. PspA4 and PspA5 are thus able to induce antibodies with a high degree of cross-reactivity in vitro, which is reflected in cross-protection of mice. We have also analyzed the contribution of the nonproline (NonPro) block within the conserved proline-rich region to the reactivity of anti-PspA antibodies, and the results indicate that N-terminal alpha-helical region, the blocks of proline repeats, and the NonPro region can influence the degree of cross-reactivity of antibodies to PspA.

A survey of nasopharyngeal carriage of penicillin nonsusceptible pneumococcal (PNSp) isolates was conducted among 1192 children attending 62 day care centers in Brazil, where pneumococcal vaccination has not been routinely introduced. Nasopharyngeal pneumococcal carriage was detected in 686 (57.6%) infants, and 178 (25.9%) of them carried PNSp isolates. Being less than 24 months of age, hospitalization in the previous 3 months, and recurrent acute otitis media were independently associated with PNSp. Serotypes 14, 23F, 19A, 6A, 6B and 19F were the most common serotype isolated accounting for 80% of the PNSp. A high proportion (35/332) of non-(sero)typeable isolates was detected, 62.9% of them PNSp. Serotypes coverage projected for the pneumococcal conjugate vaccine (PCV) 13-valent vaccine (72%) was significantly higher compared with PCV7 (58.4%) and PCV 10-valent vaccine (59.3%).

This study aimed at determining prevalence and resistance profile of Gram-negative bacilli isolated from nasopharynx of children attending day-care centers in Goiânia (Brazil). P. aeruginosa (100.0%), E. coli (50.0%), K. pneumoniae (35.3%), and E. aerogenes (16.7%) were the most frequent multi-drug resistant microorganisms isolated. No production of ESBL was detected.

Investigations regarding Staphylococcus aureus carriage among Brazilian children are scarce. We evaluated the determinants of S. aureus and methicillin-resistant S. aureus (MRSA) nasal carriage in infants attending day care centers (DCCs) and the molecular features of the MRSA strains. A total of 1,192 children aged 2 months to 5 years attending 62 DCCs were screened for S. aureus and MRSA nasal carriage. MRSA isolates were characterized by pulsed-field gel electrophoresis, multilocus sequence typing, spa typing, staphylococcal cassette chromosome (SCC) mec typing and the presence of the Panton-Valentine leukocidin gene. Logistic regression was performed to determine risk factors associated with S. aureus and MRSA colonization. S. aureus and MRSA carriage were detected in 371 (31.1%) and 14 (1.2%) children, respectively. Variables found to be independently associated with an increased risk for S. aureus carriage included being older than 24 months (odds ratio [OR], 1.8; 95% confidence interval [CI], 1.3 to 2.6) and previous DCC attendance (OR, 1.5; 95% CI, 1.0 to 2.2). Having a mother with a high level of education was a protective factor for nasal colonization (OR, 0.4; 95% CI, 0.2 to 0.8). Moreover, we observed that more children carrying MRSA had younger siblings than children not colonized by MRSA. Among the 14 MRSA strains, three SCCmec types (IIIA, IV, and V) were detected, together with a multidrug-resistant dominant MRSA lineage sharing 82.7% genetic similarity with the Brazilian clone (ST239-MRSA-IIIA; ST indicates the sequence type determined by multilocus sequence typing). Although SCCmec type V was recovered from one healthy child who had been exposed to known risk factors for hospital-associated MRSA, its genetic background was compatible with community-related MRSA. Our data suggest that DCC attendees could be contributing to MRSA cross-transmission between health care and community settings.

Objective: This study evaluated the presence of microorganisms and analyzed microscopically the pulp of 20 traumatized human teeth with intact crowns and clinical diagnosis of pulp necrosis, based on the association of at least three of the clinical criteria: crown discoloration, negative response to thermal and electric pulp vitality tests, positive response to vertical and horizontal percussion, pain on palpation or mobility., Material and Methods: Microbiological collection was performed from the root canals to evaluate the presence of microorganisms. The pulp samples were stained with hematoxylin and eosin (H.E.) for histological evaluation of possible morphological alterations., Results: Analysis of results was performed by statistical tests (linear regression test and diagnostic analysis) and subjective analysis of the sections stained with H.E. and revealed that only 15% of the sample did not exhibit microbial development. The time elapsed between dental trauma and onset of endodontic intervention ranged from 15 days to 31 months; the percussion test presented high sensitivity (80%) for detection of microorganisms in the root canal of traumatized teeth; 3 teeth (15%) did not present pulp tissue, being characterized as complete autolysis; analysis of pulp samples was performed on the other 17 cases, among which 3 (15%) exhibited partial necrosis without possibility of repair and 14 presented complete necrosis; none of the clinical criteria employed for the diagnosis of pulp necrosis in traumatized teeth was pathognomonic., Conclusions: The present results allowed the following conclusions: with regard to microbiological findings, 85% of teeth presented microorganisms in the root canal, despite the presence of an intact crown. Concerning the microscopic findings, 100% of traumatized teeth presented pulp necrosis; the pulp vitality tests based on pulp response to heat, cold and vertical percussion were the most reliable to diagnose pulp necrosis in traumatized teeth.

Coagulase-negative staphylococci are frequently associated with nosocomial infections, and healthcare professionals can be reservoirs and spread them in hospitals and in the community. The aim of this study was to identify species of coagulase-negative staphylococci isolated from the saliva of nursing professionals, determine the resistance profile and detect the mecA gene. One hundred coagulase-negative staphylococci were selected: 41 were identified as Staphylococcus epidermidis, 25 as Staphylococcus saprophyticus, 18 as Staphylococcus haemolyticus, eight as Staphylococcus cohnii, four as Staphylococcus lugdunenses, three as Staphylococcus capitis and one as Staphylococcus simulans. Of these, 32% presented oxacillin resistance, 84.4% mupirocin resistance and 32% cefoxitin resistance, and all were vancomycin sensitive. Among the oxacillin-resistant coagulase-negative staphylococci, 93.7% developed in oxacillin agar (6microg/ml) and the mecA gene was detected in 75%. The results indicate that higher investments should be directed towards identifying coagulase-negative staphylococcus species in healthcare institutions and in the community.