Genomic characterization of Salmonella enterica serovar Choleraesuis from Brazil reveals a swine gallbladder isolate harboring colistin resistance gene mcr-1.1.

Salmonella enterica serovar Choleraesuis (S. Choleraesuis) is a swine-adapted serovar associated to invasive infections in humans. In Brazil, data of strains of this serovar are scarce. In the present study, six S. Choleraesuis strains of animal (n = 5) and human (n = 1) origin from Brazil were screened for phenotypic antimicrobial resistance using disk-diffusion assay and using whole-genome sequencing data to search for antimicrobial resistance genes, plasmids, prophages, and Salmonella pathogenicity islands (SPIs). Its genetic relatedness was evaluated by MLST and SNP analysis. A single isolate from swine gallbladder harbored the colistin resistance gene mcr-1.1 into a IncX4 plasmid. In the six strains analyzed, resistance was found to tetracycline, nalidixic acid, ciprofloxacin, ampicillin, piperacillin, streptomycin, cefazoline, gentamycin, sulfamethoxazole-trimethoprim, and choloramphenicol, along with resistance genes aac(6')-Iaa, aac(3)-IV, aph(3'')-Ib, aph(6)-Id, aph(4)-Ia, aadA1, aph(3')-IIa, blaTEM-1A, floR, sul1, sul2, tet(B), drfA1, erm(B), mph(B), lnu(G), qacE, and gyrA point mutation Serine83 → Tyrosine and parC Threonine57 → Serine. Furthermore, IncF and IncH plasmids, ten SPIs, and seven prophage types were detected. All strains were assigned to ST145 and five belonged to a common SNP cluster of S. Choleraesuis strains from Brazil. The presence of S. Choleraesuis isolated from animals harboring relevant antimicrobial resistance profiles and virulence determinants reinforced the urge for enhanced surveillance to avoid its transmission to humans through food items.

Molecular epidemiology of carbapenem-resistant Acinetobacter baumannii from southern Brazil / Epidemiologia molecular de Acinetobacter baumannii resistente aos carbapenêmicos provenientes do sul do Brasil / Epidemiología molecular de Acinetobacter baumannii resistente a carbapenem del sur de Brasil

Background and Objectives: carbapenem resistance in Acinetobacter baumannii has reached extremely high levels worldwide, and class D OXA-type carbapenemases are the main associated mechanism. This study aimed to assess the phenotypic and molecular profile of clinical carbapenem-resistant A. baumannii (CRAb) isolates from a southern Brazilian border region. Methods: A. baumannii species was identified by the presence of the blaOXA-51 gene, and the susceptibility profile was determined by broth microdilution. The main carbapenemases were investigated by PCR and the molecular typing was performed by PFGE. Results: during the study, a total of 36 CRAb were recovered, of which 85.7% were from respiratory tract samples from ICU patients. High level resistance to were found in contrast to 100% of susceptibility for polymyxin B. The blaOXA-23 gene was present in 34 isolates and was the only one detected other than blaOXA-51. Molecular typing revealed the presence of four clonal strains, two of them endemic during the period of the study. Conclusion: to the best of our knowledge, our study brings the first data about resistance profile in Acinetobacter in the western border of southern Brazil and make aware of endemic clones of CRAb-producing-OXA-23 in this region of state, contributing for the construction of the national epidemiologic scenario of CRAb.(AU)

Justificativa e Objetivos: a resistência aos carbapenêmicos em Acinetobacter baumannii atingiu níveis extremamente altos em todo o mundo, e as carbapenemases do tipo OXA classe D são o principal mecanismo associado. O objetivo deste estudo foi avaliar o perfil fenotípico e molecular de isolados clínicos de A. baumannii resistentes aos carbapenêmicos (CRAb) de uma região de fronteira do sul do Brasil. Métodos: a espécie A. baumannii foi identificada através da presença do gene blaOXA-51, e o perfil de sensibilidade foi determinado por microdiluição em caldo. As principais carbapenemases foram investigadas por PCR, e a tipagem dos isolados de CRAb foi realizada por PFGE. Resultados: durante o período do estudo, 36 CRAb foram recuperados, dos quais 85,7% foram provenientes de amostras do trato respiratório de pacientes de UTI. Uma elevada resistência a aminoglicosídeos e fluoroquinolonas foi encontrada em contraste com 100% de sensibilidade a polimixina B. O gene blaOXA-23 foi encontrado em 34 isolados e foi o único detectado além do blaOXA-51. A tipagem molecular revelou a presença de quatro linhagens clonais, duas delas endêmicas ao longo do período do estudo. Conclusão: nosso estudo traz os primeiros dados sobre o perfil de resistência em Acinetobacter na fronteira oeste do sul do Brasil e alerta para a presença de clones endêmicos de CRAb produtores de OXA-23 nessa região, contribuindo para a construção do cenário epidemiológico nacional de CRAb.(AU)

Justificación y Objetivos: la resistencia a carbapenémicos en Acinetobacter baumannii ha alcanzado niveles extremadamente altos en todo el mundo y las carbapenemases OXA de clase D son el principal mecanismo asociado. El objetivo de este estudio fue evaluar el perfil fenotípico y molecular de los aislados clínicos de A. baumannii resistentes a carbapenémicos (CRAb) de una región fronteriza en el sur de Brasil. Métodos: la especie A. baumannii se identificó a través de la presencia del gen blaOXA-51 y el perfil de sensibilidad se determinó por microdilución en caldo. Las principales carbapenemasas fueron investigadas por PCR y la tipificación se hizo con PFGE. Resultados: durante el período de estudio, se recuperaron 36 CRAb, 85,7% de muestras del tracto respiratorio de pacientes de la UCI. Se encontró una alta resistencia a los aminoglucósidos y las fluoroquinolonas en contraste con 100% de sensibilidad a polimixina B. El gen blaOXA-23 se encontró en 34 aislamientos y fue el único detectado además de blaOXA-51. La tipificación molecular reveló la presencia de cuatro cepas clonales, dos de ellas endémicas durante el período de estudio. Conclusiones: hasta donde sabemos, nuestro estudio trae los primeros datos sobre el perfil de resistencia en Acinetobacter en la frontera oeste del sur de Brasil y reconoce los clones endémicos de CRAb productores de OXA.(AU)

Emergent multidrug-resistant nontyphoidal Salmonella serovars isolated from poultry in Brazil coharboring blaCTX-M-2 and qnrB or blaCMY-2 in large plasmids.

The number of foodborne gastroenteritis caused by nontyphoidal Salmonella (NTS) worldwide is estimated to be 80.3 million each year. Currently, antimicrobial-resistant NTS disseminated in the animal environment increases the risk of aggravated foodborne outbreaks. Poultry are important source of foodborne NTS infections. This study was conducted to evaluate the phenotypic and genotypic characteristics of 83 NTS isolates from poultry, classified within 36 different serovars. The most prevalent serovar was S. Schwarzengrund (10/83), from which 8/10 were multidrug resistant (MDR). The antimicrobial susceptibility testing showed a total of 18 MDR isolates, from which 8/18 coharbored blaCTX-M-2 and qnrB5. The genes qnrB5, blaCTX-M-2, qnrB2, or blaCMY-2 were also found alone in other MDR isolates. All resistance genes were harbored in large plasmids, ranging from 30 to 270 kb. The pColE replicon was present in 8 MDR isolates; however it was not associated with resistance. ISCR1 and class I integron structures were always associated with blaCTX-M-2.

Endemicity of the High-Risk Clone Klebsiella pneumoniae ST340 Coproducing QnrB, CTX-M-15, and KPC-2 in a Brazilian Hospital.

The dissemination of multiresistant Klebsiella pneumoniae carbapenemase (KPC)-2-producing Klebsiella pneumoniae isolates belonging to international high-risk clones poses a major health care threat. In this study, 48 nonduplicated, carbapenem-resistant K. pneumoniae isolated from 2011 to 2014 in a tertiary hospital were investigated. The blaKPC-2 gene was the only determinant for carbapenem resistance. The blaCTX-M-15 gene was the main determinant for the production of extended-spectrum beta-lactamase (ESBL), whereas aph(3')-Ia and qnrB were the most common genes associated with resistance to aminoglycosides and quinolones, respectively. Nine different sequence types (STs) were identified. The most common was ST340. Molecular typing by enterobacterial repetitive intergenic consensus-PCR placed 48 strains among 10 different clusters. In the studied hospital, the high-risk clone of KPC-2-producing K. pneumoniae ST340, harboring genes that codify aminoglycoside modifying enzymes, QnrB and CTX-M-15 plus CTXM-2-type ESBLs, is disseminated and acts as a major agent of infections in critically ill patients.

Extended-spectrum cephalosporin-resistant Escherichia coli isolated from chickens and chicken meat in Brazil is associated with rare and complex resistance plasmids and pandemic ST lineages.

Objectives: Brazil is the greatest exporter of chicken meat (CM) in the world. It is of utmost importance to monitor resistance to extended-spectrum cephalosporins (ESCs) in this sector because resistance to ESCs in Escherichia coli isolated from food-producing animals may contaminate humans through the food chain. Thus, the aim of this study was to characterize and compare ESC-resistant E. coli isolated from chickens and retail CM produced in south-eastern Brazil. Methods: Five CM samples and 117 chicken cloacal swabs (CCSs) were inoculated on MacConkey agar supplemented with cefotaxime. Presumptive E. coli colonies were identified and antimicrobial susceptibility was tested. Virulence and acquired blaESBL and blaAmpC genes were sought and genetic environments characterized. Isolates were typed by phylogenetic grouping, XbaI-PFGE and MLST. Results: All five CM samples and 36 CCSs (30.8%) were positive for the presence of ESC-resistant E. coli, leading to the selection of 58 resistant isolates. ESC resistance was mostly due to the presence of the chromosome-encoded blaCTX-M-2 gene, but plasmid-mediated blaCTX-M-2, blaCTX-M-8, blaCTX-M-15, blaCTX-M-55 and blaCMY-2 were also detected. Multireplicon plasmids were sporadically identified, such as IncHI2/P-blaCTX-M-2 and IncFII/N-blaCTX-M-55. Phylogroup D predominated, while PFGE and MLST revealed a high genetic diversity. Conclusions: Live Brazilian chickens and CM act as reservoirs of ESC-resistant E. coli and resistance genes are located on highly diverse genetic determinants. Potentially pathogenic strains, which may represent a threat to human health and a source of environmental contamination, were also identified. Active surveillance is therefore essential in Brazil's chicken production line.

A practical molecular identification of nonfermenting Gram-negative bacteria from cystic fibrosis

Abstract Identification of nonfermenting Gram-negative bacteria (NFGNB) of cystic fibrosis patients is hard and misidentification could affect clinical outcome. This study aimed to propose a scheme using polymerase chain reaction to identify NFGNB. This scheme leads to reliable identification within 3 days in an economically viable manner when compared to other methods.

Identification and characterization of plasmid-mediated quinolone resistance determinants in Enterobacteriaceae isolated from healthy poultry in Brazil.

The expression of plasmid-mediated quinolone resistance (PMQR) genes confers low-level quinolone and fluoroquinolones resistance alone. However, the association to chromosomal resistance mechanisms determines an expressively higher resistance in Enterobacteriaceae. These mechanisms are horizontally disseminated within plasmids and have contributed to the emergence of bacteria with reduced susceptibility or resistant to therapies worldwide. The epidemiological characterization of PMQR dissemination is highly relevant in the scientific and medical context, to investigate the dissemination within enterobacteria, from different populations, including humans and food-producing animals. In the present study, 200 Enterobacteriaceae isolates were harvested from poultry with cloacal swabs and identified as Escherichia coli (90.5%), Escherichia fergusonii (5.5%), Klebsiella oxytoca (2.5%) and Klebsiella pneumoniae (1.5%). Among isolates evaluated, 46 (23%) harboured PMQR genes including qnrB (43/200), qnrS (2/200) and aac(6')-Ib-cr (1/200). All isolates carrying PMQR genes showed multidrug-resistance phenotype. The 36 E. coli isolates showed 18 different PFGE types. All E. fergusonii isolates showed the same PFGE type. The two Klebsiella oxytoca belonged to two different PFGE types. The phylogenetic groups A, B1, and D were found among the E. coli harboring PMQR genes. Based on the phylogenetic analysis and PFGE, the population structure of E. coli isolates was diverse, even within the same farm. All isolates carrying qnrB and qnrS genes also harboured ColE-like plasmids. The Southern blot hybridization using the S1-PFGE revealed that the qnrB genes were located on low molecular weight plasmids, smaller than 10Kb. Resistance plasmids were sequenced and showed 100% identity with plasmid pPAB19-3. The association of PMQR genes with mobile genetic elements, such as transferable plasmids, favours the selection and dissemination of (fluoro) quinolones resistant bacteria among food-producing animals, and may play an important role in the current increased prevalence of resistant bacteria in different environments reported worldwide.

A practical molecular identification of nonfermenting Gram-negative bacteria from cystic fibrosis.

Identification of nonfermenting Gram-negative bacteria (NFGNB) of cystic fibrosis patients is hard and misidentification could affect clinical outcome. This study aimed to propose a scheme using polymerase chain reaction to identify NFGNB. This scheme leads to reliable identification within 3 days in an economically viable manner when compared to other methods.

Antimicrobial susceptibility of Salmonella Gallinarum and Salmonella Pullorum isolated from ill poultry in Brazil / Suscetibilidade a antimicrobianos de Salmonella Gallinarum e Salmonella Pullorum isolados de aves doentes no Brasil

ABSTRACT: Salmonella Gallinarum (S. Gallinarum) and Salmonella Pullorum (S. Pullorum) are poultry host-specific, agents of fowl typhoid and pullorum disease, respectively. These biovars cause septicemic infections, resulting in high mortality. Outbreaks are frequently reported worldwide, causing losses due to the elimination of infected flocks and treatments. The use of antimicrobial agents is frequent in poultry farms to prevent or treat gastrointestinal infections. In the present research it was evaluated the antimicrobial susceptibility of 50 S. Gallinarum and S. Pullorum isolates, from outbreaks that occurred between 1987 to 1991 and 2006 to 2013. The comparison of the susceptibility profiles showed that all isolates were susceptible to β-lactams. All isolates from 1987-1991 were susceptible to all antibiotics tested except NAL and CIP (78%). The susceptibility profile of S. Gallinarum (2006 - 2013 period) was the following NAL (58%), CIP (63%), ENR (67%), TET (92%), FFC (96%) and SXT (96%). S. Pullorum isolates (2006 - 2013 period) showed the following susceptibility rates to NAL (65%), CIP (71%), ENR (94%) and TET (94%). All isolates were susceptible to β-lactams tested, however, resistance to quinolones and fluoroquinolones increased over time. Furthermore, low levels of resistance to other antibiotics were found in recent isolates, such as tetracyclines.

RESUMO: Salmonella Gallinarum (S. Gallinarum) e Salmonella Pullorum (S. Pullorum) são patógenos hospedeiro-específico de aves, agentes do tifo aviário e pulorose, respectivamente. Estes biovares causam infecções septicêmicas, resultando em alta mortalidade. Surtos são frequentemente relatados em diversos países, causando prejuízos devido à eliminação de lotes infectados e tratamentos. Agentes antimicrobianos são utilizados frequentemente em granjas avícolas para prevenir ou tratar infecções gastrointestinais. No presente trabalho, foi avaliada a susceptibilidade antimicrobiana de 50 isolados de S. Gallinarum e S. Pullorum, obtidos durante surtos que ocorreram entre 1987 a 1991 e entre 2006 a 2013. A comparação dos perfis de sensibilidade mostrou que todas as amostras são sensíveis aos β-lactâmicos. Todos os isolados de 1987-1991 foram sensíveis a todos os antibióticos testados, exceto NAL e CIP (78%). O perfil de susceptibilidade de S. Gallinarum (surtos de 2006 a 2013) foi NAL (58%), CIP (63%), ENR (67%), TET (92%), FFC (96%) e SXT (96%). Isolados de S. Pullorum (surtos de 2006 a 2013) apresentaram as seguintes taxas de sensibilidade: NAL (65%), CIP (71%), ENR (94%) e TET (94%). Todas as amostras foram sensíveis ao β-lactâmicos testados, no entanto, a resistência às quinolonas e fluoroquinolonas aumentou ao longo do tempo. Além disso, baixos níveis de resistência a outros antibióticos foram encontrados em isolados recentes, tais como as tetraciclinas.

Evaluation of polymorphisms in pbp4 gene and genetic diversity in penicillin-resistant, ampicillin-susceptible Enterococcus faecalis from hospitals in different states in Brazil.

The aim of the present study was to verify whether penicillin-resistant, ampicillin-susceptible Enterococcus faecalis (PRASEF) occurred in Brazil prior to the beginning of the 21st century, and to verify whether ampicillin susceptibility can predict susceptibility to other ß-lactams in E. faecalis with this inconsistent phenotype. The presence of polymorphisms in the pbp4 gene and genetic diversity among the isolates were investigated. Of 21 PRASEF analyzed, 5 (23.8%) and 4 (19.0%) were imipenem and piperacillin resistant simultaneously by disk diffusion and broth dilution respectively, contradicting the current internationally accepted standards of susceptibility testing. Sequencing of pbp4 gene revealed an amino acid substitution (Asp-573→Glu) in all PRASEF isolates but not in the penicillin-susceptible, ampicillin-susceptible E. faecalis. Most PRASEF (90.5%) had related pulsed-field gel electrophoresis profiles, but were different from other PRASEF described to date. Results demonstrate that penicillin-resistant, ampicillin-susceptible phenotype was already a reality in the 1990s in E. faecalis isolates in different Brazilian states, and some of these isolates were also imipenem- and piperacillin-resistant; therefore, internationally accepted susceptibility criteria cannot be applied to these isolates. According to pbp4 gene sequencing, this study suggests that a specific amino acid substitution in pbp4 gene found in all PRASEF analyzed is associated with penicillin resistance.

Prior oropharyngeal colonization and ventilator-associated pneumonia.

This study evaluated the relationship between previous colonization of the oropharynx and development of ventilator-associated pneumonia through the classification of genomic fingerprint pattern by pulsed-field gel electrophoresis of both oxacillin-resistant and oxacillin-susceptible Staphylococcus aureus isolates obtained from hospitalized patients in an intensive care unit.

Penicillin-resistant, ampicillin-susceptible Enterococcus faecalis of hospital origin: pbp4 gene polymorphism and genetic diversity.

Despite the spread of penicillin-resistant, ampicillin-susceptible Enterococcus faecalis (PRASEF) isolates in diverse countries, the mechanisms leading to this unusual resistance phenotype have not yet been investigated. The aim of this study was to evaluate whether polymorphism in the pbp4 gene is associated with penicillin resistance in PRASEF isolates and to determine their genetic diversity. E. faecalis isolates were recovered from different clinical specimens of hospitalized patients from February 2006 to June 2010. The ß-lactam minimal inhibitory concentrations (MICs) were determined by E-test®. The PCR-amplified pbp4 gene was sequenced with an automated sequencer. The genetic diversities of the isolates were established by PFGE (pulsed-field gel electrophoresis) and MLST (multilocus sequencing typing). Seventeen non-producing ß-lactamase PRASEF and 10 penicillin-susceptible, ampicillin-susceptible E. faecalis (PSASEF) strains were analyzed. A single-amino-acid substitution (Asp-573→Glu) in the penicillin-binding domain was significantly found in all PRASEF isolates by sequencing of the pbp4 gene but not in the penicillin-susceptible isolates. In contrast to the PSASEF isolates, a majority of the PRASEFs had similar PFGE profiles. Six representative PRASEF isolates were resolved by MLST into ST9 and ST524 and belong to the globally dispersed clonal complex 9 (CC9). In conclusion, it appears quite likely that the amino acid alteration (Asp-573→Glu) found in the PBP4 of the Brazilian PRASEF isolates may account for their reduced susceptibility to penicillin, although other resistance mechanisms remain to be investigated.

Prior oropharyngeal colonization and ventilator-associated pneumonia

This study evaluated the relationship between previous colonization of the oropharynx and development of ventilator-associated pneumonia through the classification of genomic fingerprint pattern by pulsed-field gel electrophoresis of both oxacillin-resistant and oxacillin-susceptible Staphylococcus aureus isolates obtained from hospitalized patients in an intensive care unit.

Molecular characterization of Klebsiella pneumoniae carbapenemase-producing isolates in southern Brazil.

Carbapenem-resistant Enterobacteriaceae have been frequently reported worldwide. They represent a serious concern because of the limited therapeutic options. The aim of this study was to investigate the molecular epidemiology of 14 Klebsiella pneumoniae carbapenemase (KPC) producers among 345 clinical isolates of Enterobacteriaceae with reduced susceptibility to carbapenems recovered from 11 separate hospitals in southern Brazil. The blaKPC-2 gene was detected in 14 isolates (4 %): six Enterobacter cloacae, five K. pneumoniae and three Serratia marcescens. Most of these isolates exhibited high-level resistance against ß-lactams and ciprofloxacin, while the most active drugs were polymyxin B and amikacin. Genetic environment analysis, based on the classical Tn4401 structure, revealed six distinct platforms. Plasmids carrying blaKPC-2 were not typable and most were approximately 20 kb. Only KPC carbapenemases were found among the isolates studied, highlighting the local relevance of these enzymes in acquired resistance to carbapenems in Enterobacteriaceae. Our results contribute to the understanding of carbapenem resistance in Enterobacteriaceae and to the molecular characterization of KPC-2-producing isolates in Brazil.

Comparison of three molecular typing methods to assess genetic diversity for Mycobacterium tuberculosis.

This study describes the comparison of three methods for genotyping of Mycobacterium tuberculosis, namely MIRU-VNTR (mycobacterial interspersed repetitive units-variable number of tandem repeats), spoligotyping and, for the first time, MLST (Multilocus Sequence Typing). In order to evaluate the discriminatory power of these methods, a total of 44 M. tuberculosis isolates obtained from sputum specimens of patients from Brazil were genotyped. Among the three methods, MLST showed the lowest discriminatory power compared to the other two techniques. MIRU-VNTR showed better discriminatory power when compared to spoligotyping, however, the combination of both methods provides the greatest level of discrimination and therefore this combination is the most useful genotyping tool to be applied to M. tuberculosis isolates.

Dissemination of blaKPC-2 by the spread of Klebsiella pneumoniae clonal complex 258 clones (ST258, ST11, ST437) and plasmids (IncFII, IncN, IncL/M) among Enterobacteriaceae species in Brazil.

This article reports the spread of bla(KPC-2) in the Sao Paulo and Rio de Janeiro states, facilitated by globally spread K. pneumoniae clonal complex 258 (CC258) clones (ST258, ST11, and ST437) and a diversity of plasmids (IncFII, IncN, and IncL/M, two untypeable plasmids carrying Tn4401a or Tn4401b) successfully disseminated among species of the Enterobacteriaceae (Enterobacter cloacae, Serratia marcescens, and Citrobacter freundii). It also constitutes the first description of sequence type 258 (ST258) in Brazil, which was associated with a nosocomial hospital outbreak in Ribeirao Preto city.

First report of vancomycin-resistant Enterococcus faecalis in Uberaba, Minas Gerais state

In this study we report the first isolation of VanA-type vancomycin-resistant Enterococcus faecalis strains from two different patients hospitalized in the same intensive care unit at the hospital of Universidade Federal do Triângulo Mineiro (UFTM), Uberaba, Minas Gerais, Brazil.

Staphylococcus hominis subsp. novobiosepticus strains causing nosocomial bloodstream infection in Brazil.

OBJECTIVES: To report the isolation of six Staphylococcus hominis subsp. novobiosepticus (SHN) strains from hospitalized patients with bloodstream infections in two Brazilian hospitals and to characterize their susceptibility profile to several antimicrobials. METHODS: Species identification was performed by biochemical methods and sodA gene sequencing. The MICs of antimicrobials were determined by broth and agar dilution methods and by Etest. Isolates were typed by PFGE and PCR amplification was used to detect the ccr gene complex and the mec class. Morphometric evaluation of cell wall was performed by transmission electron microscopy (TEM). RESULTS: Susceptibility profiles indicated that the majority of isolates (five) were multidrug-resistant. Overlapping and multiplex PCR showed that five out of the six strains harboured SCCmec type III with class A mec and type 3 ccr. The initial vancomycin MIC value of 4 mg/L for these strains increased to 16-32 mg/L after growth for 10 days in BHI broth supplemented with this antimicrobial. TEM indicated that vancomycin resistance was associated with cell wall thickening and to another mechanism not fully elucidated. Only one SHN strain was oxacillin- and vancomycin-susceptible. The nosocomial infections in at least five of the patients from both hospitals were caused by a single clone of SHN. CONCLUSIONS: It is very important to consider SHN strains as the cause of nosocomial infections. The clinical implications resulting from the pattern of multidrug resistance in these strains may be complicated by the emergence of vancomycin resistance.

Molecular characterization of enterococci harboring genotype and phenotype incongruence related to glycopeptide resistance isolated in Brazilian hospitals.

Three Enterococcus faecalis and one Enterococcus faecium strains were characterized by plasmid profile, pulsed-field gel electrophoresis (PFGE) and determination of antimicrobial minimal inhibitory concentrations. VanA elements were characterized by Long PCR, overlapping PCR and DNA sequencing. Enterococcal strains showed resistance to vancomycin and harbored the vanA gene, and three these were teicoplanin susceptible while one showed intermediate resistance to teicoplanin. Two E. faecalis strains showed indistinguishable PFGE profile while the third was unrelated. E. faecalis strains showed a deletion in the right terminal region of the Tn1546-like element. The E. faecium strain showed an insertion element in the vanXY intergenic region. Mutations in VanA elements were not found. Rearrangements in the VanA element could be responsible for incongruities in genotype and phenotype in these strains.

Molecular characterization of enterococci harboring genotype and phenotype incongruence related to glycopeptide resistance isolated in Brazilian hospitals

Three Enterococcus faecalis and one Enterococcus faecium strains were characterized by plasmid profile, pulsed-field gel electrophoresis (PFGE) and determination of antimicrobial minimal inhibitory concentrations. VanA elements were characterized by Long PCR, overlapping PCR and DNA sequencing. Enterococcal strains showed resistance to vancomycin and harbored the vanA gene, and three these were teicoplanin susceptible while one showed intermediate resistance to teicoplanin. Two E. faecalis strains showed indistinguishable PFGE profile while the third was unrelated. E. faecalis strains showed a deletion in the right terminal region of the Tn1546-like element. The E. faecium strain showed an insertion element in the vanXY intergenic region. Mutations in VanA elements were not found. Rearrangements in the VanA element could be responsible for incongruities in genotype and phenotype in these strains.