text:
Religion: not determined yet
Name: Lorraine Miller
Classification: supporting character
First Appearance: The Human Torch (vol. 1) #30 (May 1948): "The Hyena Strikes!"
Creators: William Woolfolk, Alex Schomburg
Number of Appearances: 1
Enemy of: Mr. Lennox, The Hyena
Defended or Helped by: Human Torch, Toro
Occupation: business owner
Worked for: Miller Trucking Company
Gender: female
Note: led trucking co. after father killed by Hyena
Suggested links for further research about this character:
- http://www.comicboards.com/marvelguide/m2.html
- http://www.marvunapp.com/master/miemim.htm
- http://www.marvunapp.com/master/lehlh.htm
- http://www.marvunapp.com/master/hvhz.htm
- https://www.comics.org/issue/6725/
- http://comicbookdb.com/issue.php?ID=46939


text:
Verified email at id.uff.br
Associate Professor, Department of Microbiology and Parasitology, Instituto Biomédico
Bacteriology, Gram-positive cocci, antimicrobial resistance, molecular epidemiology
Scopus Publications
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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
Infection, Genetics and Evolution, ISSN: 15671348, eISSN: 15677257, Published: June 2020 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.
Felipe P.G. Neves, Nayara T. Cardoso, Claudete A.A. Cardoso, Lúcia M. Teixeira, and Lee W. Riley
Vaccine, ISSN: 0264410X, eISSN: 18732518, Pages: 5265-5269, Published: 23 August 2019 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
BMC Infectious Diseases, eISSN: 14712334, Published: 29 May 2019 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.
Patricia Alice Knupp-Pereira, Nayara Torres Cardoso Marques, Lúcia Martins Teixeira, Helvécio Cardoso Corrêa Póvoa, and Felipe Piedade Gonçalves Neves
Brazilian Journal of Microbiology, ISSN: 15178382, eISSN: 16784405, Published: 2019 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.
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 in Microbiology, eISSN: 1664302X, Issue: MAR, Published: 2019 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
Journal of Antimicrobial Chemotherapy, ISSN: 03057453, eISSN: 14602091, Pages: 1206-1212, Published: 1 May 2018 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
Acta Virologica, ISSN: 0001723X, Pages: 172-178, Published: 2018 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
Infection, Genetics and Evolution, ISSN: 15671348, eISSN: 15677257, Pages: 56-62, Published: November 2017 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
Epidemiology and Infection, ISSN: 09502688, eISSN: 14694409, Volume: 145, Pages: 1720-1726, Published: 1 June 2017 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
Vaccine, ISSN: 0264410X, eISSN: 18732518, Pages: 2794-2800, Published: 15 May 2017 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
Scientific Reports, eISSN: 20452322, Published: 28 March 2017 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
PLoS Neglected Tropical Diseases, ISSN: 19352727, eISSN: 19352735, Published: 23 March 2017 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
Diagnostic Microbiology and Infectious Disease, ISSN: 07328893, eISSN: 18790070, Pages: 281-285, Published: 1 March 2017 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.
Ledy Horto Santos Oliveira, Larissa Silva Santos, Carolina Oliveira Silva, Everton Faccini Augusto, and Felipe Piedade Gonçalves Neves
Brazilian Journal of Infectious Diseases, ISSN: 14138670, eISSN: 16784391, Pages: 88-91, Published: 1 January 2017 Elsevier BV
Human papillomavirus (HPV) has been found in several regions of the body, including the oral cavity. Recently, this virus has been associated with oropharyngeal cancer, but little is known about HPV transmission to the oral cavity. We carried out a study to investigate concurrent oral and cervical infections in 76 asymptomatic women attending a healthcare program. Demographic and behavior data were obtained through a structured questionnaire. Oral and cervical mucosa scrapings were collected and stored for DNA extraction. HPV DNA amplification was performed by polymerase chain reaction assay (PCR) using both primers My09/My11 and FAP59/64, followed by HPV typing with restriction fragment length polymorphism analysis (RFLP) and sequencing. The data collected revealed no risk factors for HPV infection in these 76 women. HPV prevalence of 9.2 and 5.3% was found in cervical and oral mucosa, respectively. Concurrent infections by discordant types were detected in one case only. Sequencing procedures allowed us to detect a new putative HPV 17 subtype from the Betapapillomavirus genus. Our results support the view that cervical and oral HPV infections are independent events. The observed low prevalence of both oral and cervical HPV infections could be associated with attendance in a healthcare program.
Muniqui S. Capett, Patricia Vollú-Silva, Vanessa A. Melchiades, Luciana C. Bokehi, Fernanda M. Araújo, Ianick Souto Martins, Felipe P. G. Neves, Alice G. M. Gonzalez, Eric Oswald, Geraldo R. de Paula, and Lenise A. Teixeira
Current Microbiology, ISSN: 03438651, eISSN: 14320991, Pages: 624-632, Published: 1 November 2016 Springer Science and Business Media LLC
The objective of this work was to assess the genetic characteristics of uropathogenic Escherichia coli, ciprofloxacin resistance or susceptibility, obtained from patients with gynecological cancer and urinary tract infection (UTI). Seventy-seven E. coli ciprofloxacin-resistant isolates and 38 ciprofloxacin-susceptible were analyzed by polymerase chain reaction (PCR) to determine the phylogenetic groups, virulence factors as iucC, fyuA, hlyC, cnf1 genes, and pks pathogenicity island. The presence of genes related to ciprofloxacin resistance such as qnrA, qnrB, qnrS, aac(6′)-Ib-cr, and qepA, and the sequencing of DNA gyrase genes and topoisomerase IV were determined. The genetic profile of the isolates was determined by pulsed-field gel electrophoresis (PFGE). Statistical analysis was performed using Fisher's exact test and Chi-square test. Phylogenetic group B2 was the most prevalent although a great genetic diversity was observed by PFGE. Only genes associated to siderophores were found in ciprofloxacin-resistant isolates; however, in ciprofloxacin-susceptible isolates, genes related to siderophores and toxin, were detected. Additionally qnrB was detected in both populations, ciprofloxacin resistant and susceptible. DNA mutations in gyrA were Ser-83-Leu and Asp-87-Asn and in parC were Ser-80-Ile and Glu-84-Val, Glu-84-Lys. In conclusion, it was observed a high prevalence of qnrB in the population studied; in addition, it was the first time the pks island was observed only in ciprofloxacin-susceptible isolates.
M.S. Xavier, C.N. Fonseca, R.F. Ferreira, T.X. Castro, F.P.G. Neves, A.V. Barbosa, A.M.F. Cerqueira, and N.R.P. Almosny
Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, ISSN: 01020935, eISSN: 16784162, Pages: 1197-1200, Published: 2015 FapUNIFESP (SciELO)
This report aimed to study the interference in molecular testing for Ehrlichia canis and Anaplasma platys in blood of 155 dogs from the coastal region of Rio de Janeiro. Five Anaplasmataceae positive samples but negative for E. canis and A. platys, from microfilaremic animals, were chosen for sequencing. These sequences, when compared to Gen et Bank database, showed 88% to 100% similarity with Wolbachia spp. denoting an interference in the detection of DNA from other members of Anaplasmataceae, possibly due to a high concentration of Wolbachia spp. DNA.
Glauber P Arêas, Rôde BB Schuab, Felipe PG Neves, and Rosana R Barros
Memorias do Instituto Oswaldo Cruz, ISSN: 00740276, eISSN: 16788060, Volume: 109, Pages: 935-939, Published: 1 November 2014 FapUNIFESP (SciELO)
Streptococcus pyogenes is responsible for a variety of infectious diseases and immunological complications. In this study, 91 isolates of S. pyogenes recovered from oropharynx secretions were submitted to antimicrobial susceptibility testing, emm typing and pulsed-field gel electrophoresis (PFGE) analysis. All isolates were susceptible to ceftriaxone, levofloxacin, penicillin G and vancomycin. Resistance to erythromycin and clindamycin was 15.4%, which is higher than previous reports from this area, while 20.9% of the isolates were not susceptible to tetracycline. The macrolide resistance phenotypes were cMLSB (10) and iMLSB (4). The ermB gene was predominant, followed by the ermA gene. Thirty-two emm types and subtypes were found, but five (emm1, emm4, emm12, emm22, emm81) were detected in 48% of the isolates. Three new emm subtypes were identified (emm1.74, emm58.14, emm76.7). There was a strong association between emm type and PFGE clustering. A variety of PFGE profiles as well as emm types were found among tetracycline and erythromycin-resistant isolates, demonstrating that antimicrobial resistant strains do not result from the expansion of one or a few clones. This study provides epidemiological data that contribute to the development of suitable strategies for the prevention and treatment of such infections in a poorly studied area.
Renata Fernandes Ferreira, Aloysio de Mello Figueiredo Cerqueira, Tatiana Xavier de Castro, Eliane de Oliveira Ferreira, Felipe Piedade Gonçalves Neves, André Victor Barbosa, Daniel de Barros Macieira, and Nádia Regina Pereira Almosny
Revista Brasileira de Parasitologia Veterinaria, ISSN: 0103846X, Pages: 301-308, Published: 2014 FapUNIFESP (SciELO)
The aim of this study was to characterize Ehrlichia canis strains from naturally infected dogs in Rio de Janeiro, Brazil. In addition, all the clinical and hematological findings observed in these dogs were reported. PCR targeting the 16S rRNA gene was used for diagnostic purposes, and the TRP19 and TRP36 genes were sequenced to evaluate the genetic diversity. Fifteen samples were positive for E. canis. The polymerase chain reaction for the TRP19 gene resulted in 11 amplicons (11/15), which were cloned into the pGEM-T easy vector for sequencing. The complete sequence of TRP19 gene was compared to those in the GenBank, revealing high identicalness. Phylogenetic analysis on the TRP36 gene sequences demonstrated two distinct strains from two dogs, named 56C and 70C. The 56C strain was grouped with the strain Cuiaba 16, which is a hybrid strain formed by Brazilian and US genogroups; and the 70C strain was grouped with other strains of the US genogroup, thus suggesting that there are at least two genogroups of E. canis in Rio de Janeiro (US and Brazilian). Those animals, in which the 70C and 56C strains were isolated, showed distinct clinical and hematological manifestations of 1the disease. The appearance of different genotypes may express new phenotypes, thus resulting in different forms of presentation of the disease and making its diagnosis more complex.
Danielle Ferreira Lima, Nathalia Brito Veloso Brazão, Tania Wrobel Folescu, Felipe Piedade Neves, Alex Guerra Ferreira, Erica Aparecida Santos, Elizabeth Andrade Marques, and Robson Souza Leão
Diagnostic Microbiology and Infectious Disease, ISSN: 07328893, eISSN: 18790070, Pages: 59-62, Published: January 2014 Elsevier BV
The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) is increasing in patients with cystic fibrosis (CF). We report a molecular characterization, antimicrobial resistance, and Panton-Valentine leukocidin (PVL) toxin gene detection of MRSA strains from 28 Brazilian pediatric CF patients (1 strain per patient). A significant proportion (50%) of MRSA SCCmec IV isolates was observed. Nearly half of MRSA strains harboring the PVL genes distributed in all SCCmec types detected. Multilocus sequence typing (MLST) analyses showed majority (57.1%) of the isolates belonged to known epidemic lineages, such as UK/EMRSA-3, Pediatric/USA 800, Southwest Pacific clone, and Brazilian/Hungarian clone. To our knowledge, this is the first Brazilian study of molecular epidemiology based on MLST and SCCmec typing and the first description of PVL genes in MRSA from CF patients.
P. H. C. Candido, L. d. S. Nunes, E. A. Marques, T. W. Folescu, F. S. Coelho, V. C. N. de Moura, M. G. da Silva, K. M. Gomes, M. C. d. S. Lourenco, F. S. Aguiar, F. Chitolina, D. T. Armstrong, S. C. Leao, F. P. G. Neves, F. C. d. Q. Mello, and R. S. Duarte
Journal of Clinical Microbiology, ISSN: 00951137, eISSN: 1098660X, Pages: 2990-2997, Published: August 2014 American Society for Microbiology
Worldwide, nontuberculous mycobacteria (NTM) have become emergent pathogens of pulmonary infections in cystic fibrosis (CF) patients, with an estimated prevalence ranging from 5 to 20%. This work investigated the presence of NTM in sputum samples of 129 CF patients (2 to 18 years old) submitted to longitudinal clinical supervision at a regional reference center in Rio de Janeiro, Brazil. From June 2009 to March 2012, 36 NTM isolates recovered from 10 (7.75%) out of 129 children were obtained. Molecular identification of NTM was performed by using PCR restriction analysis targeting the hsp65 gene (PRA-hsp65) and sequencing of the rpoB gene, and susceptibility tests were performed that followed Clinical and Laboratory Standards Institute recommendations. For evaluating the genotypic diversity, pulsed-field gel electrophoresis (PFGE) and/or enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) was performed. The species identified were Mycobacterium abscessus subsp. bolletii (n = 24), M. abscessus subsp. abscessus (n = 6), Mycobacterium fortuitum (n = 3), Mycobacterium marseillense (n = 2), and Mycobacterium timonense (n = 1). Most of the isolates presented resistance to five or more of the antimicrobials tested. Typing profiles were mainly patient specific. The PFGE profiles indicated the presence of two clonal groups for M. abscessus subsp. abscessus and five clonal groups for M. abscesssus subsp. bolletii, with just one clone detected in two patients. Given the observed multidrug resistance patterns and the possibility of transmission between patients, we suggest the implementation of continuous and routine investigation of NTM infection or colonization in CF patients, including countries with a high burden of tuberculosis disease.
Viviane C. Souza, Fabíola C. O. Kegele, Selma R. Souza, Felipe P. G. Neves, Geraldo R. de Paula, and Rosana R. Barros
Scandinavian Journal of Infectious Diseases, ISSN: 00365548, eISSN: 16511980, Pages: 780-785, Published: October 2013 Informa UK Limited
BACKGROUND Streptococcus agalactiae is known to be the major cause of neonatal infections and also causes complications during pregnancy. METHODS One hundred and six strains of Streptococcus agalactiae recovered from clinical specimens of newborns (n = 18) and pregnant women (n = 88) were submitted to antimicrobial susceptibility testing and investigation of genetic determinants of macrolide resistance, capsular type, and virulence factors. Genetic diversity was evaluated by pulsed-field gel electrophoresis (PFGE) analysis. RESULTS Strains were susceptible to ceftriaxone, levofloxacin, penicillin G, and vancomycin and resistant to tetracycline (85.8%) and erythromycin (4.7%). Erythromycin-resistant strains presented iMLSB phenotype, harbored the ermA gene, and were closely related by PFGE. Both bac and bca genes were found in low frequencies. PFGE analysis yielded 11 DNA restriction profiles among 35 selected isolates. The major clonal group, designated as A, was composed predominantly of strains belonging to capsular type Ia. Clonal group B was composed predominantly of strains with capsular type V, including all erythromycin-resistant isolates. CONCLUSIONS Although low levels of erythromycin resistance have been observed, this is a fact of concern because this phenotype also confers resistance to clindamycin, an alternative agent for intrapartum prophylaxis. Despite the diversity of capsular types, Ia and V were among the most common and were significantly associated with distinct clonal groups. In a few cases, different capsular types were clustered into a single clonal group, which may be related to capsular switching.
T. C. A. Pinto, A. R. V. Souza, S. E. C. M. de Pina, N. S. Costa, A. A. Borges Neto, F. P. G. Neves, V. L. C. Merquior, C. A. G. Dias, J. M. Peralta, and L. M. Teixeira
Journal of Clinical Microbiology, ISSN: 00951137, eISSN: 1098660X, Pages: 3242-3249, Published: October 2013 American Society for Microbiology
Optochin (Opt) susceptibility is used largely for the identification of Streptococcus pneumoniae in diagnostic laboratories. Opt-resistant (Opt(r)) S. pneumoniae isolates have been reported, however, indicating the potential for misidentification of this important pathogen. Point mutations in the atpC gene have been associated with the emergence of Opt(r) S. pneumoniae, but data on the characterization of such atypical variants of S. pneumoniae are still limited. The present report describes the results of a polyphasic approach to identifying and characterizing 26 Opt(r) S. pneumoniae isolates recovered from patients or carriers living in Brazil. Sixteen isolates consisted of heterogeneous populations, and 10 isolates were homogeneously Opt(r). The isolates had different serotypes and antimicrobial susceptibility profiles. They also presented diverse genetic characteristics, as indicated by pulsed-field gel electrophoresis (PFGE), multilocus variable-number tandem-repeat analysis (MLVA), and pspA gene typing. Except for Opt MICs (4- to 64-fold higher among Opt(r) variants), Opt(r) and Opt-susceptible (Opt(s)) subpopulations originating from the same culture had identical characteristics. Sequencing of the atpC gene of the Opt(r) variants revealed 13 different nucleotide changes distributed among eight different codons. Changes in codon 49 were the most frequent, suggesting that this might be a hot spot for optochin resistance-conferring mutations. On the other hand, five novel types of mutations in the atpC gene (Met13Ile, Gly18Ser, Gly20Ala, Ala31Val, and Ala49Gly) were identified. In silico prediction modeling indicated that the atpC gene mutations corresponded to alterations in the transmembrane region of the ATPase, leading to a higher hydrophobicity profile in α-helix 1 and to a lower hydrophobicity profile in α-helix 2.
Felipe Piedade Gonçalves Neves, Tatiana Castro Abreu Pinto, Mariane Alves Corrêa, Roberta dos Anjos Barreto, Laís de Souza Gouveia Moreira, Havana Gomes Rodrigues, Claudete Araújo Cardoso, Rosana Rocha Barros, and Lúcia Martins Teixeira
BMC Infectious Diseases, eISSN: 14712334, Published: 13 July 2013 Springer Science and Business Media LLC
BackgroundStreptococcus pneumoniae remains a major cause of childhood morbidity and mortality worldwide. Nasopharyngeal colonization plays an important role in the development and transmission of pneumococcal diseases, and infants and young children are considered to be the main reservoir of this pathogen. The aim of this study was to evaluate the rates and characteristics associated with nasopharyngeal carriage, the distribution of serotypes and the antimicrobial resistance profiles of Streptococcus pneumoniae among children in a large metropolitan area in Brazil before the introduction of the 10-valent pneumococcal conjugate vaccine.MethodsBetween March and June 2010, nasopharyngeal swabs were collected from 242 children aged <6 years attending one day care center and the emergency room of a pediatric hospital. Pneumococcal isolates were identified by conventional methods and serotypes were determined by a sequential multiplex PCR assay and/or the Quellung reaction. The antimicrobial susceptibilities of the pneumococci were assessed by the disk diffusion method. MICs for erythromycin and penicillin were also performed. Erythromycin resistance genes were investigated by PCR.ResultsThe overall colonization rate was 49.2% and it was considerably higher among children in the day care center. Pneumococcal carriage was more common among day care attenders and cohabitants with young siblings. The most prevalent serotypes were 6B, 19F, 6A, 14, 15C and 23F, which accounted for 61.2% of the isolates. All isolates were susceptible to clindamycin, levofloxacin, rifampicin and vancomycin. The highest rate of non-susceptibility was observed for sulphamethoxazole-trimethoprim (51.2%). Penicillin non-susceptible pneumococci (PNSP) accounted for 27.3% of the isolates (MICs of 0.12-4 μg/ml). Penicillin non-susceptibility was strongly associated with serotypes 14 and 23F. Hospital attendance and the presence of respiratory or general symptoms were frequently associated with PNSP carriage. The two erythromycin-resistant isolates (MICs of 2 and 4 μg/ml) belonged to serotype 6A, presented the M phenotype and harbored the mef(A/E) gene.ConclusionsCorrelations between serotypes, settings and penicillin non-susceptibility were observed. Serotypes coverage projected for the 10-valent pneumococcal conjugate vaccine was low (45.5%), but pointed out the potential reduction of PNSP nasopharyngeal colonization by nearly 20%.
Karla Rodrigues Miranda, Felipe Piedade Gonçalves Neves, Joaquim dos Santos-Filho, Geraldo Renato de Paula, Leandro Araújo Lobo, Walter Martin Roland Oelemann, and Regina Maria Cavalcanti Pilotto Domingues
Anaerobe, ISSN: 10759964, eISSN: 10958274, Pages: 58-61, Published: February 2013 Elsevier BV
In the past few years, many studies revealed a remarkable genetic variability in Bacteroides fragilis species, and the existence of two divisions was proposed according to presence or absence of the cfiA (metallo-β-lactamase/carbapenemase) gene. The aim of this study was to evaluate the use of DNA sequence analysis for glutamate dehydrogenase (gdh), phosphoglucomutase (pgm) and esterase (est) metabolic genes, in comparison to RNA polymerase β subunit (rpoB) and 16S ribosomal RNA (rrs) gene sequencing, to identify the presence of these two groups in seventeen B. fragilis strains. Based on phylogenetic trees, only the est gene sequences generated a classification similar to rrs- and rpoB-genes. On the other hand, the genes pgm and gdh did not allow the discrimination of these divisions. The est gene sequence can be suggested as an additional tool for differentiation of the two groups in B. fragilis, providing highly reproducible and reliable data in B. fragilis taxonomy.
Vânia Lúcia Carreira Merquior, Adriana Rocha Faria, Filomena Soares Pereira da Rocha, Jaqueline Martins Morais, Felipe Piedade Gonçalves Neves, and Lúcia Martins Teixeira
Memorias do Instituto Oswaldo Cruz, ISSN: 00740276, eISSN: 16788060, Volume: 107, Pages: 557-560, Published: June 2012 FapUNIFESP (SciELO)
Here we describe the detection and characterisation of three isolates of vancomycin-resistant VanB-type Enterococcus faecalis. Sequence analysis suggested that these isolates harboured the vanB1 gene. The isolates were susceptible to the majority of antimicrobial agents tested, with the exception of chloramphenicol, erythromycin and vancomycin, and showed distinct profiles of high-level resistance to aminoglycosides. Analysis of the clonal relatedness of the vanB E. faecalis isolates showed similar pulsed-field gel electrophoresis profiles. To our knowledge, this is the first report of the occurrence of enterococcal strains carrying vanB genes in Brazil.
Ledy H.S. Oliveira, Larissa S. Santos, Everton F. Augusto, and Felipe P.G. Neves
Infection, Genetics and Evolution, ISSN: 15671348, eISSN: 15677257, Pages: 71-76, Published: January 2012 Elsevier BV
Human papillomavirus type 53 (HPV 53), which belongs to genus Alpha, species A6, has spread among women worldwide. Although it is classified as a probably high risk type, the association between HPV 53 and the development of neoplastic cervical disease is unclear, and HPV 53 is known to be genomically diverse. We investigated 15 cases of HPV 53 genital infection in women living in the state of Rio de Janeiro that were not associated with severe intraepithelial cervical neoplasia. To trace HPV 53 variants in this geographic area, we characterized the L1, E6 and E7 genes from these isolates, and undertook a phylogenetic analysis based on multiple alignment of their L1 sequences. After amplification and sequence analysis, we identified seven different L1-E6-E7 variants and a L1 co-infected isolate, which taken together had base pair changes at 29 different positions. The co-infected sample presented overlapped peaks at two positions. We also detected two new E6 genomic variants. Several base pair changes in the E6 region resulted in amino acid changes, three of which were non-conservative. The E7 gene was the most conserved sequence among those studied; in contrast, the E6 sequence reached a maximum difference of 2.79%. None of the HPV 53 isolates corresponded to the reference type. Dichotomical branching characteristic of HPV 53 was observed in all of the trees constructed, as well as in the concatenated phylogenetic tree. Probably, these variants pointing to evolutionary process, but they not appear to keep an increasing of pathogenesis Despite the limited number of samples analyzed in our work, we noticed that the same variant was found in more than one woman. Therefore, it is possible that such variants have been circulating in the female population in the state of Rio de Janeiro for a longer time or that due to host or genetic viral factors, these variants can spread more rapidly than others.
Natalia Lopes Pontes Iorio, Roberta Ferreira Caboclo, Milena Borgo Azevedo, Ariane Guimarães Barcellos, Felipe Piedade Gonçalves Neves, Regina Maria Cavalcanti Pilotto Domingues, and Kátia Regina Netto dos Santos
Diagnostic Microbiology and Infectious Disease, ISSN: 07328893, eISSN: 18790070, Pages: 32-40, Published: January 2012 Elsevier BV
Staphylococcus epidermidis is a leading cause of hospital-acquired infections, mostly associated with the use of medical devices in seriously ill or immunocompromised patients. Currently, the characteristics of methicillin-resistant S. epidermidis (MRSE) isolates from Rio de Janeiro hospitals are unknown. In this study, staphylococcal chromosomal cassette mec (SCCmec) types, antimicrobial susceptibility profiles, biofilm formation genes, and multilocus sequence types (MLST) were investigated in 35 MRSE clinical isolates. The collection of isolates was previously well characterized by pulsed-field gel electrophoresis (PFGE) into 2 main genotypes (A and B, 22 isolates) and 10 sporadic genotypes (13 isolates). MLST revealed a total of 8 different sequence types (STs), but ST2 and ST23, which were icaAB-positive, represented the majority (71.4%) of MRSE isolates tested. Almost all isolates (91.4%) belonged to clonal complex 2. SCCmec types III and IV were identified among 71.4% of the isolates, while the remaining was nontypeable. The predominant MRSE genotypes were defined as SCCmec type III/ST2 (PFGE type A) and SCCmec type IV/ST23 (PFGE type B) isolates, which were both associated with high antimicrobial resistance and presence of biofilm-related genes.
A. A. Uehara, E. L. T. Amorin, M. d. F. Ferreira, C. F. Andrade, M. B. M. Clementino, I. de Filippis, F. P. G. Neves, T. d. C. A. Pinto, L. M. Teixeira, M. Giambiagi-deMarval, and S. E. L. Fracalanzza
Journal of Clinical Microbiology, ISSN: 00951137, eISSN: 1098660X, Pages: 4208-4212, Published: December 2011 American Society for Microbiology
Despite the rapid spread of antibiotic resistance among gonococci worldwide, limited reports are available from Brazilian locations. In the present study, 25 quinolone-resistant Neisseria gonorrhoeae (QRNG) strains isolated in Rio de Janeiro, Brazil, were characterized by phenotypic and molecular methods, including analysis of mutations in the gyrA and parC genes. They represented 16.5% of the N. gonorrhoeae isolates obtained during a survey performed from 2006 to 2010. A trend for increasing resistance to ciprofloxacin was observed in the period investigated. The most prevalent pattern of mutation observed among QRNG isolates, Ser-91 to Phe and Asp-95 to Gly in gyrA and Ser-87 to Arg in parC, was detected in 40% of the isolates exhibiting MICs ranging from 4 to >32 μg/ml. Rare types of mutations were found in the gyrA gene (Gln-102 to His [12%] and Asp-95 to Tyr [4%]) and in the parC gene (Ser-88 to Thr [4%]). The genetic relationship of the QRNG isolates, evaluated by pulsed-field gel electrophoresis, suggested that the increase in the frequencies of the QRNG isolates in Rio de Janeiro, Brazil, may have arisen as a result of simultaneous spread of two clonal groups. The results also indicate that fluoroquinolones may no longer be used as first line antibiotics for the treatment of gonorrhea in Rio de Janeiro, and that programs for antimicrobial susceptibility surveillance of N. gonorrhoeae should also be implemented in other regions of Brazil.
Priscila AM Nakamura, Rôde Beatriz B Schuab, Felipe PG Neves, Cláudio FA Pereira, Geraldo R de Paula, and Rosana R Barros
Memorias do Instituto Oswaldo Cruz, ISSN: 00740276, eISSN: 16788060, Volume: 106, Pages: 119-122, Published: March 2011 FapUNIFESP (SciELO)
In this study, 100 clinical isolates of Streptococcus agalactiae recovered from genitourinary tract specimens of non-pregnant individuals living in Rio de Janeiro were submitted for antimicrobial susceptibility testing, detection of macrolide resistance genes and evaluation of the genetic diversity of erythromycin-resistant isolates. By agar diffusion method, all isolates were susceptible to ceftazidime, penicillin and vancomycin. Isolates were resistant to levofloxacin (1%), clindamycin (5%), erythromycin (11%) and tetracycline (83%) and were intermediated to erythromycin (4%) and tetracycline (6%). Erythromycin-resistant and intermediated isolates presented the following phenotypes: M (n = 3), constitutive macrolide-lincosamide-streptogramin B (MLS B, n = 5) and inductive MLS B (n = 7). Determinants of macrolide resistance genes, erm and mef, were detected in isolates presenting MLS B and M phenotypes, respectively. Randomly amplified polymorphic DNA profiles of erythromycin-resistant isolates were clustered into two major groups of similarity.
PLoS Pathogens, ISSN: 15537366, eISSN: 15537374, Pages: 1-16, Published: April 2010
Brazilian Journal of Infectious Diseases, ISSN: 14138670, Pages: 54-59, Published: Jan. / Feb. 2010
Felipe P.G. Neves, Rachel L. Ribeiro, Rafael S. Duarte, Lucia M. Teixeira, and Vânia L.C. Merquior
International Journal of Antimicrobial Agents, ISSN: 09248579, Pages: 211-215, Published: March 2009 Elsevier BV
We describe the characteristics of seven unusual isolates of vancomycin-resistant enterococci (VRE) carrying both the vanC1 and vanA genes that were detected during a 3-month survey carried out to investigate the occurrence of faecal carriage of VRE. The isolates were identified as Enterococcus gallinarum and showed high-level resistance to both vancomycin and teicoplanin (minimum inhibitory concentrations >256 microg/mL and 64-96 microg/mL, respectively). All seven isolates were also resistant to chloramphenicol, erythromycin and high levels of gentamicin, and showed intermediate susceptibility to both quinolones tested (ciprofloxacin and norfloxacin). Susceptibility to fosfomycin, rifampicin and tetracycline varied among isolates. High-level resistance to gentamicin was associated with the aac(6')-aph(2'') gene, and resistance to erythromycin was associated with the erm(B) gene. The seven vanA-carrying E. gallinarum isolates had similar pulsed-field gel electrophoresis (PFGE) profiles. The emergence of multiple antimicrobial resistance, including high-level resistance to glycopeptides, among E. gallinarum points out the need to increase awareness for detection and proper characterisation of these microorganisms, as they may represent potential reservoirs of transmissible, clinically significant resistance genes in nosocomial settings.
V. L. C. Merquior, F. P. Goncalves Neves, R. L. Ribeiro, R. S. Duarte, E. de Andrade Marques, and L. M. Teixeira
Journal of Medical Microbiology, ISSN: 00222615, Pages: 244-245, Published: February 2008 Microbiology Society
A case of a post-surgical patient who developed a fatal bloodstream infection caused by high-level vancomycin-resistant Enterococcus gallinarum is reported. The isolate was found to carry both the vanC1 and vanA genes. This is the first report of an invasive infection associated with a vanA E. gallinarum isolate in Brazil.

subset: commoncrawl


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I. INTRODUCTION. This is the first segment of a two-part article on measuring the performance of law enforcement agencies.
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Manchester City eSports is delighted to announce the two sleeve partners for its first bespoke eSports kit, Etisalat and Nexen Tire.
The partnership will see Etisalat and Nexen Tire support Manchester City's professional FIFA players, Shaun "Shellzz" Springette & Ryan Pessoa during the FIFA 20 Global Series season, including being present for all live events.
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Ryan and Shellzz' first matches in the new kit will kick off around 2.45pm CET today. You can follow them on FIFA's channels including FIFA.com, Facebook, Twitter, Twitch and Youtube.


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Beautiful quality sparkling diamond Tiara to compliment her hair for any special occasion. Available in 3 different sizes at different prices. Small $4.99 : 3.0cm X 2.0cm (QAT-01) Medium $14.99 : 7.0cm X 4.0cm (QAT-02) XLarge $24.99 : 10.0cm X 5.0cm (QAT-04) Regular Price: $24.99
Flower Girls Basket Creme/White/Pink QL-01QL - 01 $28.00
Flower girl's basket, beautifully wrapped with pink silky satin. Hand stitched pearls and ribbons to finish. Available in Creme, White and Pink.
FLOWER-01 Beautiful Silky Satin Carnation Hair Clip and Brooch in Many ColoursFLOWER-01
Beautiful satin carnation hair clips are the perfect accessories for your little one's outfit. Can also be pinned onto dress as a decorative brooch. Many colours available for all your matching needs. Flowers are approximately 7cm in diameter.
Headband with Jacquard Print QHB-55GDFREEQHB-55GDFREE $18.00
Pretty girls headband with gold jacquard design. It looks so simple but sweet. Matching with our girls winter jacquard dress QWDS815. Price for one headband only.
Princess For the Day Tiara Headband QAT-12QAT-12 $28.00
Pretty girls headband with Tiara. Silver-tone finish. Crystal embellishments. Set on a slim band. Flexible arms for an adjustable fit.
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QAH-06 Beautiful Flower Crown in White or IvoryQAH-06 $16.00
Sweet and elegant flower crown for your little one. The perfect accompaniment to any formal dress. Adorned with satin roses and pretty pearls. Two sash leading off the back. Comes in White or Ivory.
QAH-07 Beautiful Wedding Style Veil mid-length in White or IvoryQAH-07
Classic elegant veil for your little one. The perfect accompaniment to any formal dress. Mid-length, goes just below the shoulders. Pretty satin rose adorns the top. Comb fastening. Comes in White or Ivory.
QHB-52W Pure White Headband Bow for Girls Rose Swirl TextureQHB-52
Pretty girls headband with huge bow, rose swirl material. Simple, elegant and sweet. Perfect for pairing with our selection of pure white formal dresses. Quality guaranteed. You won't be disappointed with our products. Combine headbands to your order to save on postage fees. Price for one headband only.
UQBG Baby Girl Cute Flower Silky Carnation HeadbandQBG
Cute baby girl headband with silky carnation flower design. Lovely lace elastic band cotton made silky soft for your little darlings. Comes in an assortment of colours, the white one is ideal for baptism. Quality guaranteed. Price is only for 1 headband. Add headbands to your order of shoes/clothes and save on postage fee!


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Office Hours 10:30a - 8:00p Monday - SundayJune 1 - Sep 7, 2020
Centennial Campground is open for the 2020 season by reservation only. Advanced reservations can be made online, day of reservations can be made by calling 343-6986. No walk-in camping will be allowed at this time. Day of reservations can be made until 7pm.
Our regular showers and bathroom are currently closed due to COVID-19 precautions. We're working to open them when possible. We have four regularly cleaned portable toilets for use at the campground.
The Municipality of Anchorage offers rustic camping opportunities during the summer months at Centennial Campground. Find yourself surrounded by the forest right in the heart of Anchorage. Visit our historic log cabin office, toast some s'mores over the fire, and let us welcome you to Alaska.
Centennial Campground, located in Centennial Park, offers 84 camp sites; 21 RV only sites with electric (sites 1-21), 20 RV only sites without electricity, 42 RV or Tent sites without electricity and 2 group sites.
Centennial Park is located less than 15 minutes from Downtown Anchorage and is close to hiking trails, bus lines, restaurants and other recreational activities.
You can reserve online at ActiveNet. Reservations will be accepted 180 days in advance and sites close for online reservations 1 days before the date of arrival.
For the benefit of all campers we ask that patrons obey the following rules:
Overnight camping is allowed, with a maximum stay of 14 days. Camping spots are on a first come first serve and reservation basis.
For more information call (907) 343-6986.
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8400 Starview Dr.
Phone: 343-6986 Fax: 337-0158
e-mail: firstname.lastname@example.com
Assistant Manager: Will Daye-mail: firstname.lastname@example.com
Recreation Supervisor: Zach Moss
e-mail: firstname.lastname@example.com


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A breakdown of how each country and continent voted on 'The Best' award
The Best 2019 Messi beat Van Dijk by eight points
Leo Messi picked up The Best FIFA Men's Player award in Milan on Monday night, surpassing Virgil van Dijk by eight votes and Cristiano Ronaldo by ten.
The Dutch defender was the initial favourite but the Barcelona superstar ended up with the most acclaim from his peers.
Just off podium were Liverpool forwards Mohamed Salah and Sadio Mane who occupied the fourth and fifth positions respectively.
Further down in terms of votes were Kylian Mbappe, Frenkie de Jong, Eden Hazard, Matthijs de Ligt and Harry Kane.


text: Right after researching relevant articles and performing content material evaluation on entertainment web sites for my senior project, it was concluded that broadcasts containing data potentially dangerous to the well getting of celebrities was present in both mainstream to entertainment-focused sources. Her fast wit keeps the show moving rapidly and she has build a sturdy Tv following. He is best recognized for his major roles in the Tv series What Happened in Bali (2004), That Winter, the Wind Blows (2013), and It's Okay, That is Really like (2014), as effectively as the films A Dirty Carnival (2006) and A Frozen … Read More


text: I woke up this Monday with a stiff and sore neck. I must've slept in an awkward position or something, because my neck was hurting even if I tried turning it slowly. I was like a Robogirl! So, I called in sick and went to see my physical therapist who recommended R.I.C.E. therapy and some low-impact stretches which really helped ...Read More »


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PCV system is causing it. You need to add a catch can in the line and drain it regularly.
Sent from my iPhone using Tapatalk
what is causing oil to collect in my supercharger. I have an 01 silver lightning
PCV system is causing it. You need to add a catch can in the line and drain it regularly.
Sent from my iPhone using Tapatalk
94 Black Lighting Freshly rebuilt and modded to my liking!
If you voted for Obama in 08 to prove you werent a racist
You better vote for someone else in 2012 to prove your not an Idiot.:oldtu:
Well I see we are a nation of IDIOTS!!!
The Navy seals eliminated one Muslim threat to this country its up to the voters to eliminate the other!
Must be a nation of Muslims as well!
The Gen. II Lightnings are known for this. The PCV system dumps any residual oil into the plenum behind the IAC vavle. As was said, an oil separator works to keep it out. Also there is an elbow down on the back of what is called the turkey pan, the lower intake underneath the intercooler. THis eventually will go bad on you and you will end up with P0171 and 174 codes for being too lean on the mixture. Most of us pull the supercharger at some point to clean the intercooler, because all that oil ends up accumulating on it. While the supercharger is off, remove the elbow from the back of the block and put a 3/8 NPT plug in from the inside using lots of sealant on it. Then you can delete the hose running up the back of the block, and put an oil separator in between the PCV valve and the plenum.
I did all that to mine last year when I swapped from the stock Eaton to a Magnum Powers blower. The catch can is one that Mustang owners use, available on Cramazon.
I have the phone number of a fellow that makes custom heater hoses for our Lightnings. THese are much shorter than the stock ones and allow you to clean up the engine bay on the passenger side for easier plug changes and just all around cleaner look. If you look at the pic below, you will not see those unsightly heater hoses intruding. Then delete the EGR valve on the driver's side, if you live in a state that doesn't do smog tests that is.


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07 Feb How To Hack Your Fitness to Get In Shape and Stay That Way For Life – AMA #96
Posted at 00:53h in Ask Me Anything 0 Comments
How To Hack Your Fitness to Get In Shape and Stay That Way For Life
Listen to the Podcast:
Live Ask Me Anything #96 – Topics Covered:
How To Hack Your Fitness to Get In Shape and Stay That Way For Life
Listen to the Podcast:
Topics Covered:
(Minute Markers for Facebook Video)
2:25- I'm obsessed with this topic right now, and I'm giving you the tools to be fit FOR LIFE in this episode
11:53- the KEYS to being fit forever
27:38- Aerobic threshold training: you've been missing this KEY piece of the puzzle
31:45- Commenting on Comments: This feels like something I can do! I feel like I should be doing something now that I've lost 50lbs.
33:43- Commenting on Comments: If you haven't gotten a custom nutrition plan, you're missing out!
35:02- Q: I think a few weeks back you said you don't recommend IF long term. I am a 'one meal a day' guy, am I missing something?
44:49- Commenting on Comments: I feel so judged by most health gurus who criticize that I'm overweight, you don't do that. Thank you for changing the world one person at a time!
48:24- Do things that you LIKE!
53:04- Commenting on Comments: Your brain and heart won't be magically fixed by losing weight. There's more work to be done to HEAL
57:40- There's a NEW 5 day Clovis challenge coming!
Links and Resources:
Join The Clovis Academy for Free!
I Am Clovis
The Perfect Paleo Powder
The Clovis Culture YouTube Channel
The Movement Fix Programs
**Full Medical Disclaimer:
**The opinions expressed in this video and by Justin Nault are published for educational and informational purposes only and are not intended as a diagnosis, treatment or as a substitute for professional medical advice, diagnosis, and treatment. Please consult a local physician or other health care professionals for your specific health care and/or medical needs or concerns.
Justin Nault, as well as any site or distribution channel where this video may appear, does not endorse or recommend any commercial products, medical treatments, diet plans (i.e. but not limited to terms like "paleo" or "keto"), pharmaceuticals, brand names, processes, or services, or the use of any trade, firm, or corporation name is for the information and education of the viewing public, and the mention of any of the above does not constitute an endorsement, recommendation, or favoring by Justin Nault, or Clovis LLC or any third party whatsoever. In most instances Justin Nault is speaking "off the cuff"; for that reason, all statements – even a statement such as "studies show" and the like – should be heavily scrutinized. Should you decide to take any advice in this video, and act on it in your own life, or suggest it to another, you do so at your own risk and detriment.**


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Ever hear of Teton Guitars from Idaho, USA? Maybe not but you've
probably heard of Taylor, Guild, Gibson, Martin etc... While this
guitar is priced far lower than those major builders the luthiers at
Teton have learned from the innovation of these guitar greats and made a
comparable product for far less. This American brand is standing on
the shoulders of the giants and applying their own flavor to new
technology and designs the other builders brought to the industry. I
was skeptical when I was told this also, but as I have played these
Tetons I have been amazed at the sound and quality. Everyone who plays
these has similar comments.
We can special order anything from this company!
Is there something you want we don't have in-stock? Just visit their site. Then get price and availability from us on anything in their entire product line. Order it from us, get an additional 10% Off "MAP" pricing & Free shipping on $100!
There are currently no products in stock. Please visit manufacturers website for special order requests.


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Sunstone Tumbles
Sunstone is like your daily dose of Vitamin C for the spirit! Everyone should add natural sunstone to their collection. It carries the radiance of the sun and glimmers when you hold it up to the light.
By stimulating the sacral chakra, Sunstone brings an abundance of light into dark corners of the mind, making it an excellent healing aid for clearing away negative thought patterns. It helps us to feel fresh and enthusiastic while increasing vitality and cleansing the aura and chakras.
Sunstone crystal healing properties evoke the notion of joie de vivre. Following the spirit of the time-honoured proverb, Sunstone crystal healing properties gives you the guidance to turn your face towards the sun, letting the shadows fall behind you. Call on the power of Sunstone and learn how to make peace with your past mistakes. After all, it's our imperfections that help us grow and transform.
Sunstone reminds us to keep a healthy balance between career demands and fun in the sun!
Stone Details
Size: 25-35mm
Origin: India
Listing is for 1 tumbled stone intuitively selected for you. Each piece is unique & one of a kind and has been cleansed with sage


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Take a deep breath and let it fly!.
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The flu has arrived in the Bay Area. Prepare for it like an epidemiologist
San Francisco Chronicle
Though this year's flu season has nothing on last year's — one of deadliest in the past decade — when it comes to influenza, vigilance is ever important, say infectious disease and public health experts.
Local and state reports show that the flu season is well under way in the Bay Area, with the first deaths announced this month. Now that the flu is again widely circulating, doctors, nurses and public health officials are closely watching several key surveillance reports to help them prepare for the season at hand.
"The flu is not making the news like last year's pandemic did. But it's definitely out there," said Dr. Jeffrey Silvers, medical director of infectious disease at Sutter Health. "We're still going to have a lot of people get sick and a lot of people die."


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I had two 150kg bakes of rice hulls delivered (more 'dumped' in the driveway where they'll get wet and I have NO chance of shifting them — thanks from nothing Brown Wigg I take 2 days off work waiting for delivery and it happens bang-on school pick up time) sorry – this post was actually supposed to be about how fabulous the rice hulls are so…[Read more]
Dreadful news Vickie. Clipping a wing to ensure they stay separate is definitely the go. I got some bird netting at Dahlsens the other day – I have about 15m left over if you want it to try to increase the boundary height.
Miawzen sounds like you have good neighbours. A quick call to your shire council will tell you exactly what the local laws are in your area. Some say no roosters, others require a permit.
I can't see about the last 10 characters on the end of every line in every post so I'm filling in the gaps and hopefully am getting it right!
BW do you think they could have been hunting mice?
I was cleaning around in my bedroom and heard strange rustling in the wardrobe I ignored it for a while then had to investigate – just a young pullet!? I…[Read more]
Mister Mister is a very sad sight but he did roost. Hens don't go near him. I'm trying to ensure he gets enough water to get a good dose of the antibiotics.
Tomorrow I'll give him a bath if hell let me do I can see properly how bad the wounds are. Right now he's all caked in blood. I'm waiting for my dad to get here to help me put up bird netting.
Nothing has been under or through the 2.5m high pen fence. It had to come from above which is why I'm thinking birds. Long time locals all say carrawongs. Apparently they do the cruelest things to calves and lambs so a rooster wouldnt be too daunting and definitely eat eggs.
Went out to feed the chooks this morning and came across a devestating sight. My lovely rooster has been absolutely savaged all over his head and neck.
He's lost an eye and I doubt he'll survive (although I think it would have occured yesterday so he has made it through one cold night).
2 eggs have gone from the nest and one egg shell was in…[Read more]
Jenoka: that is a LOT of nesting boxes. How many chooks do you have?
I'd also suggest planting Roman wormwood, tansy (throw it in the nesting boxes to keep parasites from moving in) and comfrey.
Purplehat: depending on how you want to use them you can skin instead of pluck. Doesn't take nearly as long, much less stressful — and you can get them in the freeze before rigamortis (Sp?) sets in so it's much less confronting.
Having said all that I have to admit I've never done either!
I w et to the vet to ask for cocci' meds (baycox or sulfas which should cost about $25) – but after looking at Simon the rooster left with a bill for $198.00. I'm stunned. Those Chooks had better be the healthiest on the planet in a few days. All I wanted was a little pre-spring clean out to ensure they were in tip top condition before breeding…[Read more]
Light Sussex Roos went for $65 at the last traralgon auction! The woman who owned them needed to sit down. She couldn't believe her good fortune. Maybe you'll be lucky too.
Love a happy ending. :tup:
Yes, they stand. Mine manage perfectly well in a mower catcher — even the big ones.
Have to disagree Ravyk:
Metal = good
Wood = bad
All looks pretty good to me — In fact the roosts look perfect. My guess is that in summer they'll use them — in the meantime leave them be. Just either extend roof line out a bit to ensure materials stay completely dry, or replace with new bedding material and push back far enough that it's out…[Read more]
Damp housing is a recipe for coccidicosis. I'd be doing whatever is necessary to ensure 100% dry inside the coop.
Awful news – I've never seen a prolapse but I'm sure you made the kindest decision Purplehat.
The mini warm spell has caused 2 to go clucky. Hopefully they continue to sit. My main rooster was standing about eyes shut today – I hope it's not the start of something heartbreaking. We love Simon.
Big gorgeous birds! Your local feed store will be loving you coming through the door.
You won't need to put them to bed manually for long — they'll catch on in a few more days.
If you put a ping pong ball or something similar where you want them to lay there's a fair chance they will comply and learn to lay there as their first preference.…[Read more]
Great feeling of elation to find eggs in laying boxes! Go Pullets.
All mine are laying now I'm busting from a broody. Never satisfied…
replied to the topic Quiet Chickens in the forum All things Chicken 8 years, 3 months ago
Yay! Are you hoarding them to admire and gaze at or are you eating them?
It's such a thrill to find that first egg! I never tire of it.
It's raining heavily again. Here comes another flood — everything is still sodden from the last one! My lovely veranda will be a mine field of chook mess. :sick:
Two of my girls have started sleeping in trees — I suspect they're also laying somewhere other than the nesting boxes. Never a dull day in chooky paradise!
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Discussion in 'Bugs & Feedback' started by Hedley Heap, Nov 24, 2014.
Have I had a brain fart, or has recent posts gone?
The font has changed too.
Someone got the spanners out again?
I think "New Posts" has a new function.
It doesn't but still lets you click a link to Recent Posts.
Someone's broken it.
And this new font
..so TNR...we don't like change
Should be okay again now, you might have to log out and back in to see the changes
T'was just me fettling
Separate names with a comma.