FELIPE PIEDADE GONÇALVES NEVES
@uff.br
Associate Professor, Department of Microbiology and Parasitology, Instituto Biomédico
UNIVERSIDADE FEDERAL FLUMINESE
RESEARCH INTERESTS
Bacteriology, Gram-positive cocci, antimicrobial resistance, molecular epidemiology
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Patricia Alice Knupp-Pereira, Amanda Seabra Cabral, Ítalo Moraes Dolores, Amanda Beiral da Silva, Helvécio Cardoso Correa Póvoa, and Felipe Piedade Gonçalves Neves
MDPI AG
Streptococcus pneumoniae causes serious illnesses, such as pneumonia, bacteremia, and meningitis, mainly in immunocompromised individuals and those of extreme ages. Currently, pneumococcal conjugate vaccines (PCVs) are the best allies against pneumococcal diseases. In Brazil, the 10-valent and 13-valent PCVs have been available since 2010, but the threat of antimicrobial resistance persists and has been changing over time. We conducted a systematic review of the literature with works published since 2000, generating a parallel between susceptibility data on isolates recovered from colonization and invasive diseases before and after the implementation of PCVs for routine childhood use in Brazil. This systematic review was based on the Cochrane Handbook for Systematic Reviews of Interventions and Preferred Reporting Items for Systematic Literature Reviews and Meta-Analyses (PRISMA) guidelines. Despite the inclusion of PCVs at a large scale in the national territory, high frequencies of non-susceptibility to important drugs used in pneumococcal diseases are still observed, especially penicillin, as well as increasing resistance to macrolides. However, there are still drugs for which pneumococci have a comprehensive sensitivity profile.
Beatriz O. Farias, Kaylanne S. Montenegro, Ana Paula A. Nascimento, Mariana Magaldi, Andressa S. Gonçalves-Brito, Claudia Flores, Thais C. Moreira, Felipe P. G. Neves, Kayo Bianco, and Maysa M. Clementino
Springer Science and Business Media LLC
Letícia B.D.P. Fortuna, Filipe M. Miranda, Isa M.F. Antunes, Amanda B. Silva, Amanda S. Cabral, Ítalo M. Dolores, Nayara T. Cardoso-Marques, Lúcia M. Teixeira, and Felipe P.G. Neves
Elsevier BV
Amanda Beiral da Silva, Nayara Torres Cardoso-Marques, Ítalo de Moraes Dolores, Lúcia Martins Teixeira, and Felipe Piedade Gonçalves Neves
Elsevier BV
Felipe Piedade Gonçalves Neves, Tomislav Mestrovic, and Tatiana Castro Abreu Pinto
Frontiers Media SA
COPYRIGHT © 2023 Neves, Mestrovic and Pinto. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Editorial: Drug resistance in maternal and paediatric bacterial and fungal infections: Is COVID-19 changing the landscape
Felipe P. G. Neves and Tatiana C. A. Pinto
Springer International Publishing
Gabriela Ceccon Chianca, Lívia Azeredo Alves Antunes, Pâmela Oliveira Ornellas, Felipe Piedade Gonçalves Neves, Helvécio Cardoso Corrêa Póvoa, and Natalia Lopes Pontes Póvoa Iorio
Informa UK Limited
Objective: This study aimed to search for Enterococcus faecalis in children's deep carious dentine and characterize their virulence traits.Material and Methods: Eight isolates from 15 carious molars identified by 16S rDNA species-specific PCR as E. faecalis were included. These eight isolates were subject to identification by MALDI-TOF and characterized regarding: (i) bacterial aggregation and biofilm formation on polystyrene and glass, with/without saliva, as single or dual-species (associated to Streptococcus mutans); (ii) environmental pH measurement before and after 24 h incubation; (iii) acidogenicity; (iv) gelatinase production; (v) macrophage adherence; and (vi) toxicity towards Caenorhabditis elegans. Statistical analyses were performed using two-way ANOVA/Tukey or Fisher's exact tests.Results: All isolates initially identified as E. faecalis by PCR were correctly identified as Lactobacillus by MALDI-TOF, being designated as Lactobacillus misidentified as Enterococcus (LME). These isolates produced biofilm in the presence of saliva and in the dual-species assays. Bacterial aggregation was only observed in the dual-species model. After 24 h, environmental pH dropped from 7.5 to 4.5 for seven of eight isolates, and to 4.0 in all dual-species models. LME isolates were acidogenic, none of them produced gelatinase or adhered to macrophages, but all presented toxicity towards C. elegans.Conclusions: No E. faecalis were identified in the children's caries lesions. All LME isolates presented important virulence traits, including biofilm formation and high acidogenicity, which cause enamel demineralization, that might increase the risk of dental caries in children carrying LME. Thus, the correct identification and in-depth virulence characterization of microorganisms isolated from dental caries are important to understand the dynamics of this disease.
Aline R. V. Souza, Sandrine E. C. M. de Pina, Natália S. Costa, Felipe P. G. Neves, Vânia L. C. Merquior, José Mauro Peralta, Tatiana C. A. Pinto, and Lúcia M. Teixeira
Springer Science and Business Media LLC
AbstractOptochin susceptibility testing is a major assay used for presumptive identification of Streptococcus pneumoniae. Still, atypical optochin-resistant (Optr) pneumococci have been reported and this phenotype has been attributed to nucleotide substitutions in the genes coding for the F0F1ATPase. While substitutions in the atpC gene (c-subunit of ATPase) are more common and better characterized, data on mutations in the atpA (a-subunit) are still limited. We have characterized five Optr isolates presenting alterations in the atpA (Trp206Cys in four isolates and Trp206Ser in one isolate), constituting the first report of such mutations in Brazil. Most of the Optr isolates consisted of heterogeneous populations. Except for Opt MICs and the nucleotide changes in the atpA gene, Optr and Opts subpopulations originating from the same culture had identical characteristics. In addition, we compared phenotypic and genetic characteristics of these atpA mutants with those of atpC mutants previously identified in Brazil. No structural alterations were detected among predicted proteins, regardless of mutations in the coding gene, suggesting that, despite the occurrence of mutations, protein structures tend to be highly conserved, ensuring their functionalities. Phylogenetic analysis revealed that atypical Optr strains are true pneumococci and Opt resistance does not represent any apparent selective advantage for clinical isolates.
Rodrigo Poubel Vieira de Rezende, Nayara Torres Cardoso-Marques, Larissa Alexandra Silva Rodrigues, João Paulo Chevrand Latini de Almeida, Gecilmara Salviato Pillegi, Lúcia Martins Teixeira, Evandro Mendes Klumb, and Felipe Piedade Gonçalves Neves
SAGE Publications
Sir, Patients with systemic lupus erythematosus (SLE) have higher morbidity and mortality due to pneumococcal diseases than the general population. Streptococcus pneumoniae colonization in the upper respiratory tract is a prerequisite for severe pneumococcal diseases such as pneumonia, bacteraemia, and meningitis. Thus, data on the characteristics of the main circulating strains in the community setting (e.g., antimicrobial susceptibility profiles and distribution of serotypes) will be helpful in the management of pneumococcal infections and assessing the potential coverage of commercially available pneumococcal vaccines. Despite limited information on pneumococcal colonization in adults, carriage prevalence among them varies from 1% to 21% (culture-based methods), with no available data on SLE adults. Therefore, we carried out a cross-sectional study to investigate pneumococcal carriage in this population. From June 19, 2018, to January 29, 2019, we recruited 229 SLE patients aged �18 years. The participants were regularly followed up at two university hospitals in Brazil (affiliated to Universidade Federal Fluminense [n1⁄4 49] and Universidade do Estado do Rio de Janeiro [n1⁄4 180]). Nasopharyngeal and oropharyngeal swab specimens were collected in all participants. Specimens were kept at �70�C until culture could be performed. Suspected colonies were identified based on alpha-haemolysis, optochin sensitivity, and bile solubility. Capsular types were determined by multiplex polymerase chain reaction, and antimicrobial susceptibility to nine drugs was evaluated by the disk diffusion method. Minimum inhibitory concentrations of penicillin and erythromycin were determined using M.I.C.Evaluator strips (Oxoid, Basingstoke, Hants, UK). A more in-depth description of these methodologies has been previously published. The patients’ sociodemographic and clinical characteristics are shown in Supplementary table 1. Ten (4.4%) out of 229 participants were pneumococcal carriers, yielding 11 isolates (Table 1). One participant (#155) harboured two distinct strains, and three (#46, #106, #112) had the same strain in both the nasopharynx and oropharynx. Pneumococci were isolated from the oropharynx in 9 out of 10 (90%) carriers and from the nasopharynx in 4 out of 10 (40%) carriers (p1⁄4 0.01; chi-square). Except for one patient, all carriers were judged by their attending rheumatologist to be in clinical remission. None of the studied variables was associated with pneumococcal colonization in the univariate analysis (Supplementary table 1; p> 0.05). As a limitation, accurate data on the patients pneumococcal immunisation records were lacking. Seven different serotypes/serogroups were identified, while four pneumococcal strains were non-typeable (Table 1). Among the 11 isolates, 6 (54.5%) are potentially covered by the 23-valent pneumococcal polysaccharide vaccine (PPV23), and 2 (18.2%) by the 13-valent pneumococcal conjugate vaccine (PCV). All strains were sensitive to chloramphenicol, levofloxacin, rifampicin, and vancomycin. The highest resistance frequencies were observed for penicillin and tetracycline (4/11 strains; 36.4% each). We detected 3 (27.3%) multidrug-resistant (MDR) strains, i.e., resistant to at least three antimicrobial classes. In summary, we showed that 1) the pneumococcal carriage prevalence (4.4%) in our sample of SLE adults was in line with reported colonization frequencies among non-SLE adults; 2) the addition of an oropharynx swab significantly improved the diagnostic yield of S. pneumoniae; 3) MDR pneumococcal strains were not uncommon in the community, and 4) approximately half of the isolates were potentially covered by currently marketed pneumococcal vaccines, especially the PPV23. These findings have implications on clinical practice, reinforcing the need for active epidemiological surveillance and adherence to the recommendations for pneumococcal vaccination of SLE patients. Lupus
Gabriel Rezende-Pereira, Julia P Albuquerque, Monica C Souza, Barbara A Nogueira, Marlei G Silva, Raphael Hirata, Ana L Mattos-Guaraldi, Rafael S Duarte, and Felipe P G Neves
Oxford University Press (OUP)
Abstract Background Bacterial biofilm on surfaces of mammary implants is a predisposing factor for several outcomes. Because Gram-positive bacteria are potential agents of biomaterial-associated infections (BAIs), their abilities to form biofilm on breast implants should be elucidated. Objectives The aim of this study was to evaluate biofilm formation on different mammary prosthesis surfaces by major Gram-positive bacterial pathogens involved in BAIs. Methods We initially evaluated biofilm formation on polystyrene plates with and without fibrinogen or collagen for 1 reference strain and 1 clinical isolate of Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus pyogenes. We also tested the ability of clinical isolates to form biofilm on 4 different implant surfaces: polyurethane foam and smooth, microtextured, and standard textured silicone. Biofilm structure and cell viability were observed by scanning electron microscopy and confocal laser scanning microscopy. Results All strains showed strong biofilm formation on polystyrene. After fibrinogen or collagen treatment, biofilm formation varied. With fibrinogen, reference strains of S. aureus and S. pyogenes increased biofilm formation (P < 0.05). Reference strains of all species and the clinical isolate of S. pyogenes increased biofilm formation after collagen treatment (P < 0.05). In general, S. aureus showed higher capacity to produce biofilm. Scanning electron microscopy showed that biofilm attached to all surfaces tested, with the presence of extracellular polymeric substances and voids. Viable cells were more frequent for E. faecalis and S. pyogenes. Conclusions All species produced biofilm on all prosthesis surfaces and under different conditions. Micrographies indicated thicker bacterial biofilm formation on microtextured and/or standard textured silicone by all species, except E. faecalis.
P.G. Lomardo, N.T. Cardoso-Marques, L.A. Motta, T.R.S. Aguiar, F.P.G. Neves, F. Aguiar-Alves, M.D. Calasans-Maia, V. Quinelato, and P.L. Casado
Elsevier BV
AIM
This study aimed to evaluate the prevalence of S. pneumoniae colonization in three different sites in healthy adults: nasopharynx, oropharynx and gingival sulcus.
METHODS
Two-hundred and sixty five adults, aged 20-60 years, who attended dental clinics in one public university (n = 106) and one military institution (n = 159) were enrolled in this study. Pneumococcal detection was performed by direct culture (DC) and PCR for lytA gene after a broth enrichment step. Capsular types were determined by sequential multiplex PCR.
RESULTS
We identified 18 (6.8%) pneumococcal carriers among 265 adults by PCR, but only one (0.4%) pneumococcal strain was isolated by DC method. Oropharynx (17; 6.4%) was the main source of S. pneumoniae. Colonization of gingival sulcus and nasopharynx was found in 4 (1.5%) and 2 (0.8%) adults, respectively. Nine distinct capsular types were detected from 9 adults and co-colonization with 2 serotypes was confirmed in 4 (1.5%) subjects. Factors associated with carriage were being females, low level of schooling, non-military and regular medication. We observed a low (6.8%) pneumococcal carriage prevalence, but oropharyngeal samples yielded more sensitive results, especially by the PCR-based detection methodology.
CONCLUSION
Gingival sulcus was found to be a possible reservoir for S. pneumoniae independently of the oropharynx or nasopharynx colonization.
Laura Maria Andrade de Oliveira, Eric Cordeiro-Spinetti, Felipe Piedade Gonçalves Neves, Patricia Sanae Sujii, Rachel Leite Ribeiro, Sidcley Silva de Lyra, Tatiana Castro Abreu Pinto, and Maria Letícia Bonatelli
Journal of Microbiology and Biology Education American Society for Microbiology
DivulgaMicro is a Brazilian Science Communication and Outreach project run by three young female scientists. In 2020, due to the COVID-19 pandemic, we promoted the first virtual edition of the DivulgaMicro Workshop.
Barbara Araújo dos Santos, Jessica da Silva de Oliveira, Bruna Marcela Parmanhani-da-Silva, Rachel Leite Ribeiro, Lúcia Martins Teixeira, and Felipe Piedade Gonçalves Neves
Elsevier BV
We aimed to investigate the occurrence of CRISPR elements in the genomes of vancomycin-resistant (VRE) and vancomycin-susceptible (VSE) enterococci and their association with the presence of antimicrobial resistance and virulence genes. We analyzed 180 isolates, including 91 VRE and 89 VSE. Isolates were identified by PCR or MALDI-TOF. Antimicrobial susceptibility and MICs for vancomycin were determined by the disk-diffusion method and E-test®, respectively. The presence of resistance and virulence genes, as well as CRISPR elements, was investigated by PCR. We identified 95 (53%) E. faecalis, 78 (43%) E. faecium, five (2.8%) E. gallinarum, and one (0.6% each) E. casseliflavus and E. durans. The highest and the lowest non-susceptibility frequencies were observed for erythromycin (n = 152; 84.4%) and fosfomycin (n = 5; 2.8%), respectively. Most erythromycin-resistant isolates had the erm(B) gene (106/152; 69.7%). Of 118 (65.6%) isolates with high-level resistance to aminoglycoside, 69 (58.5%) had at least one aminoglycoside resistance gene, mostly ant(6)-Ia and aac(6')-Ie + aph(2″)-Ia. We found at least one virulence gene among 135 (75%) isolates, mostly gelE (79/180; 43.9%). Ninety-two (51.1%) isolates had at least one CRISPR element, especially CRISPR3 (62/92; 67.4%). CRISPR elements were more common among E. faecalis, in which we observed a relationship between the absence of CRISPR and the presence of the vanA resistance gene, and the hyl and esp virulence genes. Among VRE. faecium, a relationship was found between the absence of CRISPR and the hyl gene. In conclusion, we found evident associations between the lack of CRISPR elements with species, multidrug resistance, and major resistance- and virulence-associated genes.
Patricia Alice Knupp-Pereira, Nayara Torres Cardoso Marques, Lúcia Martins Teixeira, Helvécio Cardoso Corrêa Póvoa, and Felipe Piedade Gonçalves Neves
Springer Science and Business Media LLC
In 2010, the 10-valent (PCV10) and 13-valent (PCV13) pneumococcal conjugate vaccines were introduced in Brazil to immunize children, resulting in serotype replacement. We analyzed 253 carriage isolates recovered from children aged <6 years in Brazil, including 124 and 129 isolates from the pre-PCV10/13 (December 2009-July 2010) and post-PCV10/13 (September-December 2014) periods, respectively, to investigate the prevalence of PspA families and pilus islets, potential vaccine candidates. Serotypes and resistance profiles were previously characterized. We used PCR to type PspA families (Fam1-3) and pilus islets (PI-1 and PI-2). We identified the PspA family of 130 (51.4%) isolates. PspA families 1, 2, and 3 were identified in 12.2%, 38.7%, and 0.4% of the isolates, respectively. Eighteen (58.1%) Fam1 isolates were serogroup 6. Nine (81.8%) of 11 serotype 14 isolates were Fam2. Fam1 isolates resistant to penicillin (50%), erythromycin (43.7%), clindamycin (31.2%), and chloramphenicol (6.2%) were only found after PCV10/13 introduction. Resistance among Fam2 isolates was higher in the post-PCV10/13 period to erythromycin (1.8% vs. 18.6%), clindamycin (0 vs. 13.9%), and tetracycline (10.9% vs. 16.3%). PI-I was detected in 42 (16.6%) isolates. Fourteen (56%) of 25 serotype 15B/C and nine (81.8%) of 11 serotype 14 isolates had PI-1 (p < 0.01). Eight (3.2%) isolates had PI-2, and six (75%) were serogroup 19. Five (2%) serogroup 19 isolates had both PI-1 and PI-2. We found associations between serogroups/serotypes, PspA families, and pilus islets, but distribution of PspA families and pilus islets was similar in both periods. After universal vaccination, we observed higher antimicrobial resistance frequencies, regardless PspA or pilus types.
Felipe P.G. Neves, Nayara T. Cardoso, Claudete A.A. Cardoso, Lúcia M. Teixeira, and Lee W. Riley
Elsevier BV
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.
Felipe Piedade Gonçalves Neves, Mariel Asbury Marlow, Gabriel Rezende-Pereira, Marcos Gabriel Pinheiro, Allyne Fandino Martinez dos Santos, Maria de Fátima Nogueira de Freitas, Rosana Rocha Barros, Fábio Aguiar-Alves, Claudete Aparecida Araújo Cardoso, and Lee Woodland Riley
Springer Science and Business Media LLC
BackgroundStaphylococcus aureus and beta-hemolytic streptococci (BHS) diseases disproportionately affect populations in middle/low-income countries. To assess if this disparity is reflected in colonization by these organisms, we compared their colonization frequency among children from different socioeconomic status (SES) communities in a city with high income inequality.MethodsBetween May–August 2014, we collected nasal and throat swabs to investigate S. aureus and BHS colonization among children who attended private and public pediatric clinics. Patients were classified as high SES, middle/low SES, and slum residents. We investigated the antimicrobial resistance profile, the SCCmec types and the presence of PVL genes among methicillin-resistant S. aureus (MRSA). We also examined the antimicrobial resistance profile and serogroups of BHS.ResultsOf 598 children, 221 (37%) were colonized with S. aureus, of which 49 (22%) were MRSA. MRSA colonization was higher in middle/low SES (n = 18; 14%) compared with high SES (n = 17; 6%) and slum (n = 14; 8%) residents (p = 0.01). All MRSA strains were susceptible to clindamycin, nitrofurantoin, and rifampin. The highest non-susceptibility frequency (42.9%) was observed to erythromycin. SCCmec type V was only found in isolates from high SES children; types I and II were found only in middle/low SES children. Ten (20%) MRSA isolates carried PVL genes. Twenty-four (4%) children were BHS carriers. All BHS (n = 8) found in high SES children and six (67%) isolates from slum patients belonged to group A. All group B streptococci were from middle/low SES children, corresponding to five (71%) of the seven BHS isolated in this group. BHS isolates were susceptible to all drugs tested.ConclusionsChildren from different SES communities had distinct bacterial colonization profiles, including MRSA carriage. Public health officials/researchers should consider SES when assessing disease transmission and control measures.
Tatiana Castro Abreu Pinto, Felipe Piedade Gonçalves Neves, Aline Rosa Vianna Souza, Laura Maria Andrade Oliveira, Natália Silva Costa, Luciana Fundão Souza Castro, Cláudia Rezende de Vieira Mendonça-Souza, José Mauro Peralta, and Lúcia Martins Teixeira
Frontiers Media SA
Streptococcus pneumoniae is a major cause of community-acquired pneumonia and meningitis, and it is also found as a commensal, colonizing the human upper respiratory tract of a portion of the human population. Its polysaccharide capsule allows the recognition of more than 90 capsular types and represents the target of the currently available pneumococcal conjugate vaccines (PCVs), such as the 10-valent (PCV10) and the 13-valent (PCV13). Penicillin non-susceptible pneumococci (PNSP) have been listed as one of the current major antimicrobial-resistant pathogen threats. In Brazil, the emergence of PNSP was initially detected in the mid 1990s and PCV10 has been part of the National Immunization Program since 2010. Here, we investigated the distribution of capsular types and penicillin susceptibility profiles of 783 pneumococcal strains isolated in Brazil between 1990 and 2014 to assess the evolution of penicillin non-susceptibility among pneumococci associated with asymptomatic carriage and invasive pneumococcal disease (IPD). The most common serotypes among carriage isolates were 19F, 6B, 6C, 23F, and 14. Among IPD isolates, the most frequent types were 14, 3, 6B, 5, 19F, and 4. We detected 21 types exclusively associated with IPD isolates, whereas non-typeable (NT) isolates were only detected in carriage. Nearly half of the isolates belonged to PCV10 serotypes, which remarkably decreased in occurrence (by nearly 50%) after PCV10 introduction (2011–2014), while non-PCV10 serotypes increased. PNSP frequency and levels were much higher among carriage isolates, but PNSP belonging to PCV10 serotypes were more common in IPD. While the occurrence of PNSP has decreased significantly among IPD isolates since 2011, it kept increasing among carriage strains. Such a difference can be attributed to the serotypes that emerged in each clinical source after PCV10 usage. PNSP with multidrug resistance profiles that emerged within carriage isolates comprised mostly serotypes 6C and 35B, as well as NT isolates. In turn, penicillin-susceptible capsular types 3, 20, and 8 have risen among IPD. Overall, our results reinforce the relevance of PNSP surveillance over a long period of time to better understand the dynamics of antimicrobial resistance in response to PCV introduction and may also contribute to improve control measures toward drug-resistant pneumococci.
Felipe P G Neves, Nayara T Cardoso, Aline R V Souza, Robert E Snyder, Mariel M Marlow, Tatiana C A Pinto, Lúcia M Teixeira, and Lee W Riley
Oxford University Press (OUP)
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.
L. OLIVEIRA, L. SANTOS, and F. NEVES
AEPress, s.r.o.
Human papillomavirus (HPV) exhibits epithelial and mucosal tropism. HPV type 17 belongs to the Betapapillomavirus genus and molecular cloning experiments have identified two subtypes (17a and 17b) isolated from epidermodysplasia verruciformis (EV). HPV subtypes are characterized by dissimilarities from 2 to 10% at the nucleotide level from their referenced HPV. The aim of this study was to characterize the L1, E6, E7 and LCR sequences from an isolate, which was recovered from the oral mucosa of an asymptomatic 63 year-old woman. The whole late gene 1 (L1) was amplified using several sets of primers. The complete early genes 6 and 7 (E6, E7) and the long control region (LCR) were amplified using specific primers. Potential binding sites for transcriptional factors within the LCR were also investigated. Within these sets, the DNA sequence was altered at 91 positions (68 in L1, 13 in E6, 8 in E7, and 2 in LCR sequences). L1 analysis showed high dissimilarity compared with the HPV 17 prototype, reaching 4% of nucleotide substitutions and leading to a probable third 17 subtype. The E6 oncoprotein presented the highest modification among the sequences studied, with four amino acid changes in comparison with the prototype isolate. One amino acid was modified at a position 62 (S-T), a zinc-binding domain (CxxC(C)29 CxxC). Our findings provide data on genetic variations seen in this genotype, reaching to dichotomic branching and pointing to an evolutionary process.
Barbara A. Santos, Jéssica S. Oliveira, Nayara T. Cardoso, André V. Barbosa, Silvana V. Superti, Lúcia M. Teixeira, and Felipe P.G. Neves
Elsevier BV
Cancer and hematological malignancies constitute major comorbidities in enterococcal infections, but little is known about the characteristics of enterococci affecting cancer patients. The aim of this study was to characterize 132 enterococcal clinical isolates obtained from cancer patients attending a Cancer Reference Center in Brazil between April 2013 and March 2014. Susceptibility to 17 antimicrobial agents was assessed by disk diffusion method. Resistance and virulence genes were investigated by PCR. Multilocus sequence typing (MLST) was performed for selected Enterococcus faecalis and Enterococcus faecium isolates. The predominant species was E. faecalis (108 isolates), followed by E. faecium (18), Enterococcus gallinarum (3), Enterococcus avium (2) and Enterococcus durans (1). Multidrug-resistant (MDR) isolates made up 44.7%, but all isolates were susceptible to fosfomycin, linezolid and glycopeptides. The most prevalent genes associated with erythromycin- and tetracycline-non susceptible isolates were erm(B) (47/71; 66.2%) and tet(M) (24/68; 35.3%), respectively. High-level resistance (HLR) to gentamicin was found in 22 (16.7%) isolates and 13 (59.1%) of them carried the aac(6')-Ie-aph(2″)-Ia gene. HLR to streptomycin was detected in 34 (25.8%) isolates, of which 15 (44.1%) isolates had the ant(6')-Ia gene. The most common virulence genes were gelE (48.9%), esp (30.5%) and asa1 (29.8%). MLST performed for 26 E. faecalis isolates revealed 18 different sequence-types (STs), with seven corresponding to novel STs (625, 626, 627, 628, 629, 630, and 635). On the other hand, nine of 10 E. faecium isolates analyzed by MLST belonged to a single clonal complex, comprised of mostly ST412, which emerged worldwide after mid-2000s, but also two novel STs (963 and 964). We detected major globally disseminated E. faecalis and E. faecium clonal complexes along with novel closely related STs, indicating the fitness and continuous evolution of these hospital-adapted lineages.
H. G. RODRIGUES, T. C. A. PINTO, R. R. BARROS, L. M. TEIXEIRA, and F. P. G. NEVES
Cambridge University Press (CUP)
SUMMARYWe performed two different approaches (broth enrichment step prior to culture (BEC) and PCR (BEPCR)) for detecting Streptococcus pneumoniae from nasopharyngeal specimens collected from 242 children aged <6 years attending one hospital (n = 140) and one childcare centre (n = 102) in a major urban area in Brazil. These specimens were collected immediately before the introduction of the 10-valent pneumococcal conjugate vaccine (PCV10) and the 13-valent vaccine (PCV13) for routine use in Brazil. Results were compared with previous findings obtained with direct culture (DC) on a selective medium. Colonisation prevalence was 58·3% (n = 141), being higher among children attending the childcare centre (62·7% vs. 55%). The culture-based methods (DC and BEC) enabled the detection of S. pneumoniae in 119 (49·2%) and 115 (47·5%) children, respectively. The PCR-based method (BEPCR) was more sensitive and 137 (56·6%) carriers were identified. Twenty-six serogroups/serotypes were identified, predominantly 6B, 19F, 14, 6A, 15C and 23F. Multiple colonisation was observed in 13 (5·4%) children. The estimated serotypes coverage of available PCVs was 40·4% for the 10-valent (included in the Brazilian immunisation programme) and 55·8% for the 13-valent (only available in private clinics). The use of robust approaches to obtain a more realistic insight about the asymptomatic carrier status is of paramount importance to estimate and assess the impact of vaccine implementation. The combination between culture-based and molecular methods constitutes a suitable strategy.
Felipe P.G. Neves, Nayara T. Cardoso, Robert E. Snyder, Mariel A. Marlow, Claudete A.A. Cardoso, Lúcia M. Teixeira, and Lee W. Riley
Elsevier BV
BACKGROUND
In 2010, the 10-valent pneumococcal conjugate vaccine (PCV10) was introduced free of charge in Brazil as part of the public immunization program. Here we investigated the carriage prevalence, colonization risk factors, capsular types, and antimicrobial resistance among pneumococcal isolates obtained from children in Brazil four years after routine PCV10 use.
METHODS
Between September and December 2014, we conducted a cross-sectional study among children<6years old who attended one public and two private clinics in Niterói, RJ, Brazil to evaluate pneumococcal nasopharyngeal carriage. Antimicrobial susceptibility and capsular types were determined for all isolates.
RESULTS
Of 522 children, 118 (22.6%) were pneumococcal carriers. Being≥2years old, attending childcare center, presenting with any symptoms, having acute or chronic respiratory disease, and residing in a slum were associated with pneumococcal carriage. The most prevalent capsular types were 6C (14.5%), 15B/C (11.5%), 11A/D (9.2%), and 6A (7.6%). PCV10 serotypes represented 2.5%. All isolates were susceptible to levofloxacin, rifampicin, and vancomycin. Penicillin non-susceptible pneumococci (PNSP) comprised 39%, with penicillin and ceftriaxone MICs ranging from 0.12-8.0μg/ml and 0.012-1.0μg/ml, respectively. The 33 (28%) erythromycin-resistant isolates (MICs of 1.5 to >256μg/ml) displayed the cMLSB (72.7%) or M (27.3%) phenotypes, harboring the erm(B) and/or mef(A/E) genes. High non-susceptibility rates (>20%) to clindamycin, erythromycin, penicillin, and tetracycline were largely explained by the prevalence of multidrug resistant (MDR) serotype 6C isolates.
CONCLUSIONS
Effects of universal childhood PCV10 use on carriage were evident, with the near elimination of PCV10 serotypes. The emergence of MDR serotype 6C isolates, however, is a concern. Ongoing surveillance to monitor serotype 6C increase in invasive diseases is warranted.
Tatiana C. A. Pinto, Natalia S. Costa, Luciana F. S. Castro, Rachel L. Ribeiro, Ana Caroline N. Botelho, Felipe P. G. Neves, Jose Mauro Peralta, and Lucia M. Teixeira
Springer Science and Business Media LLC
Streptococcus pneumoniae can be classified in more than 90 capsular types, as traditionally determined by serological methods and more recently by PCR-based techniques. Such methods, however, can be expensive, laborious or unable to accurately discriminate among certain serotypes. Therefore, determination of capsular types, although extremely important for epidemiological purposes and for estimating the impact of pneumococcal conjugate vaccines, is mainly restricted to research laboratories, being rarely performed in the clinical setting. In the present study, MALDI-TOF MS was evaluated as an alternative tool to characterize 416 pneumococcal isolates belonging to serotypes 6A, 6B, 6C, 9N, 9V or 14. For MALDI-TOF MS analysis, each isolate was submitted to an extraction protocol using formic acid and acetonitrile. Measurements were performed with a Bruker Microflex LT mass spectrometer using default parameters and generating spectra in the range of 2,000–20,000 m/z. Spectra were analyzed with the BioNumerics software v7.6. Isolates were mainly distributed according to the capsular type in a Neighbor Joining tree and serotypes investigated were successfully discriminated by the presence/absence of 14 selected biomarkers. The results suggest that MALDI-TOF MS is a promising alternative for typing pneumococcal strains, highlighting its usefulness for rapid and cost-effective routine application in clinical laboratories.
Robert E. Snyder, Claire E. Boone, Claudete A. Araújo Cardoso, Fabio Aguiar-Alves, Felipe P. G. Neves, and Lee W. Riley
Public Library of Science (PLoS)
1 Division of Epidemiology, School of Public Health, University of California Berkeley, Berkeley, California, United States of America, 2 Maternal and Child Department, School of Medicine, Universidade Federal Fluminense, Niterói, Rio de Janeiro, Brazil, 3 Pathology Program, Laboratório Universitário Rodolpho Albino, Universidade Federal Fluminense, Niterói, Rio de Janeiro, Brazil, 4 Department of Microbiology and Parasitology, Biomedical Institute, Universidade Federal Fluminense, Niterói, Rio de Janeiro, Brazil, 5 Division of Infectious Diseases and Vaccinology, School of Public Health, University of California Berkeley, Berkeley, California, United States of America
Nayara Torres Cardoso, Bárbara Araújo Santos, André Victor Barbosa, Silvana Vargas Superti, Lúcia Martins Teixeira, and Felipe Piedade Gonçalves Neves
Elsevier BV
We sought to characterize pneumococcal isolates associated with bacteremia, pneumonia and meningitis in cancer patients and to estimate the coverage of the available pneumococcal vaccines. Fifty isolates recovered from 49 patients attending a cancer reference center over a 1-year period were analyzed. The prevalent serotypes were: 23F (12%), 6A (8%), 3, 4, 20, and 23A (6% each). All isolates were susceptible to chloramphenicol, levofloxacin, rifampicin, and vancomycin. Resistance or reduced susceptibility to penicillin made up 14%, and one isolate was also intermediately resistant to ceftriaxone. The three (6%) erythromycin-resistant isolates presented the M or cMLSB phenotypes and harbored the mef(A/E) gene exclusively or along with the erm(B) gene. Twenty-two (44%) isolates were closely related to 11 international clones, being strongly associated with penicillin non-susceptibility. Combined immunization with the 13-valent conjugate and the 23-valent polysaccharide vaccines might contribute to reduce (76%) the burden of the pneumococcal infections in the population investigated.