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Introduction. Group B Streptococcus (GBS) is a human commensal bacterium that is also associated with infection in pregnant and non-pregnant adults, neonates and elderly people. Gap Statement. The authors hypothesize that knowledge of regional GBS genetic patterns may allow the use of prevention and treatment measures to reduce the burden of streptococcal disease. Aim. The aim was to report the genotypic diversity and antimicrobial sensitivity profiles of invasive, noninvasive urinary and colonizing GBS strains, and evaluate the relationships between these findings. Methodology. The study included consecutive and non-duplicated GBS isolates recovered in southern Brazil from 2015 to 2017. We performed multiple-locus variable-number tandem repeat analysis (MLVA) and PCR analyses to determine capsular serotypes and identify the presence of the resistance genes mefA/E, ermB and ermA/TR, and also antibiotic susceptibility testing. Results. The sample consisted of 348 GBS strains, 42 MLVA types were identified, and 4 of them represented 64 % of isolates. Serotype Ia was the most prevalent (42.2 %) and was found in a higher percentage associated with colonization, followed by serotypes V (24.4 %), II (17.8 %) and III (7.8 %). Serotype V was associated with invasive isolates and serotypes II and III with noninvasive isolates, without significant differences. All isolates were susceptible to penicillin. GBS 2018/ hvgA was observed in 17 isolates, with 11 belonging to serogroup III. The Hunter–Gaston diversity index was calculated as 0.879. The genes mefA/E, erm/B and erm/A/TR were found in 45, 19 and 46 isolates. Conclusion. This report suggests that the circulating GBS belong to a limited number of genetic lineages. The most common genotypes were Ia/MT12 and V/MT18, which are associated with high resistance to macrolides and the presence of the genes mefA/E and ermA/TR. Penicillin remains the antibiotic of choice. Implementation of continuous surveillance of GBS infections will be essential to assess GBS epidemiology and develop accurate GBS prevention, especially strategies associated with vaccination.

Neisseria gonorrhoeae is a major concern of public health due to its extraordinary capacity to develop and acquire resistance to different antimicrobials used to treat gonorrhoea. Limited treatment options and uncontrolled transmission have raised the need to assess the antimicrobial susceptibility profile of the isolates and to establish affordable alternatives for laboratory diagnosis.This study aimed to (i) determine the susceptibility profile of 336 clinical isolates of N. gonorrhoeae to ceftriaxone, azithromycin, ciprofloxacin, spectinomycin and gentamicin by the gold standard agar dilution method; (ii) assess the agreement among agar dilution and disc diffusion results for ciprofloxacin, azithromycin, ceftriaxone, spectinomycin and gentamicin.All isolates were susceptible to ceftriaxone and spectinomycin. The levels of resistance to azithromycin and ciprofloxacin were 3.9% and 35.1%, respectively. Intermediate susceptibility to gentamicin was observed in 19.4% of isolates. There was 100% agreement between methods for spectinomycin and ceftriaxone, 99.7% for ciprofloxacin, and 85.7% for azithromycin. For gentamicin, there was 86.3% agreement between agar dilution and disc diffusion, resulting in intermediate susceptible by one method and susceptible by the other method, defined as minor errors. The discordance among agar dilution and disc diffusion results is acceptable for ciprofloxacin, ceftriaxone and spectinomycin as per CLSI M23-Ed4.Spectinomycin and gentamicin can be considered in some cases as options for the treatment of gonorrhoea in Brazil. Disc diffusion can be an alternative method in routine testing with comparable accuracy to agar dilution.

Group BThe authors hypothesize that knowledge of regional GBS genetic patterns may allow the use of prevention and treatment measures to reduce the burden of streptococcal disease.The aim was to report the genotypic diversity and antimicrobial sensitivity profiles of invasive, noninvasive urinary and colonizing GBS strains, and evaluate the relationships between these findings.The study included consecutive and non-duplicated GBS isolates recovered in southern Brazil from 2015 to 2017. We performed multiple-locus variable-number tandem repeat analysis (MLVA) and PCR analyses to determine capsular serotypes and identify the presence of the resistance genesThe sample consisted of 348 GBS strains, 42 MLVA types were identified, and 4 of them represented 64 % of isolates. Serotype Ia was the most prevalent (42.2 %) and was found in a higher percentage associated with colonization, followed by serotypes V (24.4 %), II (17.8 %) and III (7.8 %). Serotype V was associated with invasive isolates and serotypes II and III with noninvasive isolates, without significant differences. All isolates were susceptible to penicillin. GBS 2018/This report suggests that the circulating GBS belong to a limited number of genetic lineages. The most common genotypes were Ia/MT12 and V/MT18, which are associated with high resistance to macrolides and the presence of the genes

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Mycobacterium tuberculosis (M.tb), the pathogen responsible for tuberculosis (TB) poses as the major cause of death among infectious diseases. The knowledge about the molecular diversity of M.tb enables the implementation of more effective surveillance and control measures and, nowadays, Whole Genome Sequencing (WGS) holds the potential to produce high-resolution epidemiological data in a high-throughput manner. Florianópolis, the state capital of Santa Catarina (SC) in south Brazil, shows a high TB incidence (46.0/100,000). Here we carried out a WGS-based evaluation of the M.tb strain diversity, drug-resistance and ongoing transmission in the capital metropolitan region. Resistance to isoniazid, rifampicin, streptomycin was identified respectively in 4.0% (n = 6), 2.0% (n = 3) and 1.3% (n = 2) of the 151 studied strains by WGS. Besides, resistance to pyrazinamide and ethambutol was detected in 0.7% (n = 1) and reistance to ethionamide and fluoroquinolone (FQ) in 1.3% (n = 2), while a single (0.7%) multidrug-resistant (MDR) strain was identified. SNP-based typing classified all isolates into M.tb Lineage 4, with high proportion of sublineages LAM (60.3%), T (16.4%) and Haarlem (7.9%). The average core-genome distance between isolates was 420.3 SNPs, with 43.7% of all isolates grouped across 22 genomic clusters thereby showing the presence of important ongoing TB transmission events. Most clusters were geographically distributed across the study setting which highlights the need for an urgent interruption of these large transmission chains. The data conveyed by this study shows the presence of important and uncontrolled TB transmission in the metropolitan area and provides precise data to support TB control measures in this region.

Mycobacterium tuberculosis has a complex cell wall containing mycolic acids (MA), which play an important role in pathogenesis, virulence, and survival by protecting the cell against harsh environments. Studies have shown that genes encoding enzymes involved in MA synthesis are essential to mycobacterial functionality. Here, we used whole-genome sequencing to evaluate mutations in genes related to MA metabolism in M. tuberculosis isolates from pulmonary tuberculosis patients of the Florianopolis Metropolitan Area, Santa Catarina, Brazil, and assessed associations with clinical, epidemiological, and genotypic data. The mutations Rv3057c Asp112Ala (104/151), Rv3720 His70Arg (104/151), and Rv3802c Val50Phe (105/151) were identified in about 69% of the isolates and were related to the LAM lineage. SIT 216/LAM5 (13.2%, 20/151) had the highest frequency and presented the mutations accD2 Lys23Glu, kasA Gly269Ser, mmaA4 Asn165Ser, otsB1 Asp617Asn, Rv3057c Asp112Ala, Rv3720 His70Arg, Rv3802c Val50Phe, and tgs4 Ala216Glu. All SIT 73/T isolates (6.6%, 10/151) showed a characteristic and exclusive gene mutation pattern: amiD Rv3376 3790075G > A, fbpA-aftB 4266941G > A, echA11 Asn220fs, and otsB2 Ser110Arg. SITs 20/LAM1, 64/LAM6, 50/H3, 137/X2, and 119/X1 were also related to specific mutations. SITs from the LAM lineage differed in mutation profile from those of the T, Haarlem, and X lineages. Isolates from patients who had treatment failure showed mutations that do not seem to have a pattern related to this outcome. It was possible to identify a broad repertoire of single-nucleotide polymorphisms in genes related to MA metabolism in M. tuberculosis isolates. This study also described, for the first time, the variability between different SITs/sublineages of Lineage 4 circulating in Florianopolis Metropolitan Area.

Background Neisseria gonorrhoeae (NG) has an extraordinary ability to develop resistance to all antimicrobials used for its treatment. This study analysed molecular determinants of antimicrobial resistance and NG-MAST of 153 NG isolates collected at Florianopolis metropolitan area during 2008–2016. Methods Minimal Inhibitory Concentration (MIC) was determined by agar dilution and the molecular epidemiology was evaluated by NG-MAST. Results Resistance was observed to penicillin (PEN) (26.1%), tetracycline (TET) (41.2%), ciprofloxacin (CIP) (52.3%) and azithromycin (AZT) (5.2%). All isolates were susceptible to cefixime (CFX) and ceftriaxone (CRO). However, 8.5% of isolates had MIC=0.125 mg/L for CFX, one log below the resistance cut-off point (EUCAST). β-lactamase production was detected in 12.4% of isolates and one of them carried the blaTEM-135 allele. The American or Dutch tetM gene were carried by 5.2% of the isolates. Mutations in the QRDR were observed in 87.5% of isolates resistant to CIP. NG-MAST showed 64 different sequence types (STs), including 19 novel STs. ST225, ST2992, ST1582, ST338, ST1407, ST2202 and ST6827 were most prevalent. G225 genogroup was associated with resistance to CIP (p Conclusion This study showed high rates of resistance to PEN, TET and CIP associated with persistence and dissemination of gonococcal strains resistant to CIP and plasmid resistance to PEN and TET. Despite the high resistance profile to CIP, the treatment recommended to the south region of Brazil was the association of CIP-AZT until 2017, when the national recommendation changed it to CRO-AZT. Disclosure No significant relationships.

Healthcare-associated infections (HAI) are an important public health threat with the multidrug-resistant (MDR) gram-negative bacteria (GNB) being of particular concern. Here we present the antimicrobial resistance profile of HAI-related GNB (HAIrB) isolated from patients (PT), healthcare workers (HCW) and hospital environment (HE) in a six-month screening program. From the 180 sampling points distributed in six hospital units, a total of 1,080 swabs were collected allowing the isolation of 390 HAIrB: 50.5% from HE, 42.6% from PT and 6.9% from HCW. Among the HAIrB, 32.6% were characterized as MDR and 38.7% as extended-spectrum cephalosporins resistant (ESC-R), showing no differences in the distribution between PT, HE and HCW. Carbapenem resistance (CARB-R) was detected for 17.7% of all HAIrB, being higher among Acinetobacter spp. isolates (36.5%), followed by Enterobacteriaceae (14.5%) and Pseudomonas spp. (11.8%). Except for the ICU, that revealed higher MDR, CARB-R and ESC-R rates, HAIrB-resistant profiles were similarly detected within the hospital units. Prevalence of blaKPC-like and blaCTX-M-1 β-lactamases-resistance genes was higher in K. pneumoniae and E. cloacae complex, while blaOXA-23-like and blaSPM-like were higher in A. baumannii and P. aeruginosa, respectively. This study reveals that the spreading of HAIrB within a hospital environment is higher than predicted, indicating that healthcare workers, hospital areas and equipment are key players on dissemination of MDR gram-negative bacteria and shows that an active surveillance program can provide precise understanding and direct actions towards control of HAI.

The Tuberculosis (TB) notification rates are 5 to 81 times higher in prisons worldwide when compared to the general population. The state of Santa Catarina (SC) has few epidemiological data regarding TB in prisons. The aim of this study was to evaluate the molecular epidemiology of circulating strains in prisons of SC. The study comprised 95 clinical samples from six prisons. Among the cases included, all subjects were male, predominantly caucasians, and young adults, with low education level. The positive smear in the TB diagnosis comprised 62.0% of cases. About 50% of subjects had some condition associated with TB. The Spoligotyping results showed that the most frequent lineages were LAM (50.7%), T (22.2%) and S (11.6%). The 12-loci MIRU generated 62 different genotypes. The MSTs showed evolutionary relationships between Mycobacterium tuberculosis spoligotypes from SC and evolutionary relationships between the prison isolates and studied parameters. This first study on TB in prison units of SC highlighted the predominance of SIT216/LAM5, and SIT34/S. Interestingly, his profile was found to be different from that observed in a previous study performed with the state's general population. This data shows the need for continued surveillance of episodes of TB occurring among prison inmates in an emerging country like Brazil.

Introduction: Neisseria gonorrhoeae (NG), initially highly susceptible to many antimicrobials, was recently assigned as a superbug due to its ability to develop resistance to all antimicrobials introduced for treatment during the last years. This study aimed to determine the prevalence and trends of NG antimicrobial resistance (AMR) during 2008–2016 in Grande Florianopolis, Santa Catarina, Brazil. Methods A total of 152 gonococcal isolates from urogenital specimens were submitted to Santa Luzia Medical Laboratory, Florianopolis, Brazil. All isolates were identified to the species level using MALDI-TOF. The minimum inhibitory concentration (MIC) was determined using agar dilution method to penicillin, tetracycline, ciprofloxacin, ceftriaxone, cefixime and azithromycin. Quality control was performed using NG WHO reference strains and ATCC 49226. Results All isolates were sensitive to ceftriaxone (MIC 0.001–0.06 µg/ml) and cefixime (MIC: 0.0005–0.125 µg/ml). Resistance to penicilin, tetracycline, ciprofloxacin and azithromycin were 26.3%, 40.8%, 52.0% and 5.2% respectively. Conclusion The study showed increased resistance to penicillin, tetracycline, ciprofloxacin and azithromycin. No resistance to ceftriaxone and cefixime was detected. Due to the high level ciprofloxacin resistance, the dual therapy currently recommended in Brazil for gonococcal infections (ciprofloxacin plus azithromycin) is no more an effective treatment option in Grande Florianopolis. Thus, ceftriaxone constitute the treatment option for gonococcal infections, as well as in the states of Rio de Janeiro, Sao Paulo and Minas Gerais, where regional studies have already detected high level ciprofloxacin resistance. AMR testing needs to be frequently performed to ensure the treatment effectiveness.

Background Tuberculosis (TB) control programs face the challenges of decreasing incidence, mortality rates, and drug resistance while increasing treatment adherence. The Brazilian TB control program recommended the decentralization of patient care as a strategy for combating the disease. This study evaluated the performance of this policy in an area with high default rates, comparing epidemiological and operational indicators between two similar municipalities. Methods This study analyzed epidemiological and operational indicators on new cases of pulmonary tuberculosis reported in the Brazilian Notifiable Diseases Information System between 2006 and 2015. In addition, to characterize differences between the populations of the two studied municipalities, a prospective cohort study was conducted between 2014 and 2015, in which patients with new cases of culture-confirmed pulmonary tuberculosis were interviewed and monitored until the disease outcome. A descriptive analysis, the chi-square test, and a Poisson regression model were employed to compare TB treatment outcomes and health care indicators between the municipalities. Results Two thousand three hundred nine cases were evaluated, of which 207 patients were interviewed. Over the 2006–2015 period, TB incidence per 100,000 population in the municipality with decentralized care was significantly higher (39%, 95% CI 27–49%) in comparison to that of the municipality with centralized care. TB treatment default rate (45%, 95% CI 12–90%) was also higher in the municipality with decentralized care. During the two-year follow-up, significant differences were found between patients in centralized care and those in decentralized care regarding treatment success (84.5 vs. 66.1%), treatment default (10.7 vs. 25.8%), illicit drug use (27.7 vs. 45.9%), and homelessness (3.6 vs. 12.9%). The operational indicators revealed that the proportion of control smear tests, medical imaging, and HIV tests were all significantly higher in the centralized care. However, a significantly higher proportion of patients started treatment in the early stages of the disease in the municipality with decentralized care. Conclusions These data showed a low success rate in TB treatment in both municipalities. Decentralization of TB care, alone, did not improve the main epidemiological and operational indicators related to disease control when compared to centralized care. Full implementation of strategies already recommended is needed to improve TB treatment success rates.

Salmonella enterica subsp. enterica serovar Enteritidis (SE) is currently responsible for significant losses in the poultry industry with consequences on public health. In the present study, 38 SE isolates from biological material of chicken breeders were characterized using phage typing, antimicrobial resistance profile and pulsed field gel electrophoresis (PFGE). The phage type (PT) 7 and 9 were predominant and 26.3% of the isolates were resistance to nalidixic acid (NAL). The phage typing and analysis of antimicrobial resistance profile showed high discriminatory power (0.85). The PFGE profile of 13 strains with XbaI and SpeI discriminated the SE phage types, as well characterized strains from the same serovar. The results showed the importance to associate different methods for the characterization of SE and suggest that poultry products may have been source of human salmonellosis in the Parana State during the period of study. Salmonella enterica subsp. enterica sorovar Enteritidis (SE) é responsável por prejuízos significativos à avicultura, com conseqüências importantes para saúde pública. No presente estudo foi realizada a caracterização de 38 isolados de SE provenientes de material biológico de reprodutoras destinadas à produção de frangos de corte através de fagotipagem, perfil de resistência antimicrobiana e de eletrofose de campo pulsado (PFGE). Os fagotipos (PT) predominantes foram PT7 e PT9 e 26,3% dos isolados apresentaram resistência ao ácido nalidíxico (NAL). A fagotipagem e a análise do perfil de resistência antimicrobiana juntos mostraram elevado poder discriminatório (0,85). Os perfis de PFGE de 13 cepas, com as endonucleases XbaI e SpeI permitiram discriminar os fagotipos de SE, bem como associar perfis de um mesmo fagotipo. Os resultados mostram a importância da associação de métodos para a caracterização de SE e sugerem que produtos da avicultura de corte podem ser fonte de salmonelose humana.

Objective: The aim of the present study was to analyze two DNA extraction methods for use in molecular GBS diagnostics and compare them to the results of culture method. Materials and methods: Two hundred vaginal samples were collected during the antenatal period, as per CDC recommendations, and atr gene polymerase chain reaction (PCR) was performed. Results: Comparison of the two DNA extraction methods demonstrated 45% concordance. Sensitivity and specificity for 5 M Guanidine DNA extraction were 100% and 86.5%, respectively. Sensitivity and specificity for the commercial DNA extraction kit were 50% and 95%, respectively. Conclusion: This study demonstrated that 5 M Guanidine DNA extraction was superior to the commercial kit, with PCR presenting a shorter turnaround time than culture. PCR could improve sensitivity and, therefore, may be a useful screening method. Sensitive GBS diagnosis allows for an effective treatment, with decreased newborn morbidity and mortality; therefore, cost-effectiveness studies are necessary to assess the feasibility of implementing PCR in routine laboratories, together with maternity ward collaboration.

A total of 41 Salmonella Enteritidis strains, including phago-types (PTs) PT4 and PT9, were characterized by antimicrobial resistance profiles and PFGE. Of these strains, 34 were isolated from patients and foods, and 7 were of poultry origin. All strains were susceptible to ampicillin, chloramphenicol, cefotaxime, ciprofloxacin and trimethoprim/sulfamethoxazole, and 41.5 % (n = 17) were resistant to nalidixic acid. PFGE analysis using XbaI and SpeI restriction enzymes resulted in X1S1 as the prevalent pattern, which was present in 48.8 % (n = 20) of epidemic strains and in one strain isolated from discarded hatching eggs. Distinct patterns were found for the other strains isolated from poultry (X3S1, X8S8, X11S12, X11S13, X16S1 and X13S15). The S. Enteritidis PT9 strains associated with outbreaks of salmonellosis were highly similar (≥0.90), suggesting clonality. The PFGE genotypes were related to the PTs, and it was possible to differentiate strains isolated from patients with salmonellosis from other strains of non-epidemic origin. The PFGE results suggested that the S. Enteritidis strains of poultry origin were a possible source of human salmonellosis during the study period.

One-hundred sixty-eight group B streptococcal (GBS) isolates from a Brazilian hospital were phenotypically and genotypically characterized. Isolates were recovered from human sources from April 2006 to May 2008 and classified as either invasive, noninvasive, or colonizing isolates. Classical methods for serotyping and antibiotic resistance profiling were employed. Clonal groups were also defined by pulsed-field gel electrophoresis (PFGE). Results showed that susceptibility to beta-lactam antimicrobials was predominant among the isolates. Only 4.7% were resistant to erythromycin and clindamycin. The erm (B) gene was widely detected in our GBS isolates, according to our phenotypic results (constitutive macrolide-lincosamide-streptogramin B [cMLS B ] resistance phenotype), and the erm (A) gene was also detected in some isolates. MLS B resistance was restricted to strains isolated from patients with noninvasive infections and carriers. Serotype Ia was predominant (38.1%), serotype IV isolates were found at a high frequency (13.1%), and few isolates of serotype III were identified (3%). Pulsed-field gel electrophoresis results revealed a variety of types, reflecting the substantial genetic diversity among GBS strains, although a great number of isolates could be clustered into two major groups with a high degree of genetic relatedness. Three main PFGE clonal groups were found, and isolates sharing the same PFGE type were grouped into different serotypes. Furthermore, in a few cases, isolates from the same patients and possessing the same PFGE type were of different serotypes. These findings could be related to the occurrence of capsular switching by horizontal transfer of capsular genes.

Background In the last few years, carbapenem-resistant Acinetobacter baumannii isolates (CR-AB) have been identified worldwide. The first description of OXA-23–producing A baumannii in Brazil was from the city of Curitiba in 2003. The aim of the present study was to evaluate the persistence and dissemination of the first OXA-23–producing A baumannii clone isolated from patients in Hospital de Clinicas, Curitiba, Brazil. Methods An antimicrobial susceptibility profile of the isolates was determined by the standard agar dilution method. Molecular detection of β-lactamase genes was done by polymerase chain reaction. The clonal relationship of the isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Epidemiologic and clinical features were evaluated as well. Results Genotypic analysis of 172 CR-AB isolates by PFGE identified 3 distinct major PFGE clusters (A, B, and C, accounting for 36, 69, and 65 isolates, respectively). All isolates carried the bla OXA-23 –like gene and were multidrug-resistant, but were susceptible to tigecycline and polymixin B. The mortality rate related to CR-AB infection was 45.4%, and ventilator-associated pneumonia and bloodstream infections were the most frequent clinical manifestations. Conclusions The presence of 3 clones among the CR-AB isolates suggests that cross-transmission was the main mechanism responsible for dissemination of OXA-23 producers. PFGE pattern A was genotypically similar to that of the first OXA-23–producing A baumannii clone identified in Curitiba in 1999. This clone persisted in the same hospital until April 2004. The presence of the bla OXA- 23–like gene was the main mechanism associated with carbapenem resistance among the isolates studied.

Universidade Federal de Sao Paulo, UNIFESP, Disciplina Infectol, Dept Med, Sao Paulo, SP, Brazil