First Report of the Globally Disseminated IncX4 Plasmid Carrying the mcr-1 Gene in a Colistin-Resistant Escherichia coli Sequence Type 101 Isolate from a Human Infection in Brazil.

A colistin-resistant Escherichia coli strain was recovered from a patient with a diabetic foot infection in Brazil. Whole-genome analysis revealed that the E. coli isolate belonged to the widespread sequence type (ST) 101 and harbored the mcr-1 gene on an IncX4 plasmid that was highly similar to mcr-1-bearing IncX4 plasmids that were recently identified in Enterobacteriaceae from food, animal, and human samples recovered on different continents. These results suggest that self-transmissible IncX4-type plasmids may represent promiscuous plasmids contributing to the intercontinental spread of the mcr-1 gene.

Silent dissemination of colistin-resistant Escherichia coli in South America could contribute to the global spread of the mcr-1 gene.

During a Brazilian multicentric antimicrobial resistance surveillance study, colistin resistance was investigated in 4,620 Enterobacteriaceae isolated from human, animal, food and environmental samples collected from 2000 to 2016. We present evidence that mcr-1-positive Escherichia coli has been emerging in South America since at least 2012, supporting a previous report on the possible acquisition of mcr-1-harbouring E. coli by European travellers visiting Latin American countries.

Genetic background of novel sequence types of CTX-M-8- and CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae from public wastewater treatment plants in São Paulo, Brazil.

The release of extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae to the environment is a public health issue worldwide. The aim of this study was to investigate the genetic background of genes encoding ESBLs in wastewater treatment plants (WWTPs) in São Paulo, southeastern Brazil. In 2009, during a local surveillance study, seven ESBL-producing Enterobacteriaceae strains were recovered from five WWTPs and screened for ESBL genes and mobile genetic elements. Multilocus sequence typing (MLST) was carried out, and wild plasmids were transformed into electrocompetent Escherichia coli. S1-PFGE technique was used to verify the presence of high molecular weight plasmids in wild-type strains and in bla ESBL-containing E. coli transformants. Strains harbored bla CTX-M-8, bla CTX-M-15, and/or bla SHV-28. Sequencing results showed that bla CTX-M-8 and bla CTX-M-15 genes were associated with IS26. MLST revealed new sequence types for E. coli (ST4401, ST4402, ST4403, and ST4445) and Klebsiella pneumoniae (ST1574), except for one K. pneumoniae from ST307 and Enterobacter cloacae from ST131. PCR and S1-PFGE results showed CTX-M-producing E. coli transformants carried heavy plasmids sizing 48.5-209 kb, which belonged to IncI1, IncF, and IncM1 incompatibility groups. This is the first report of CTX-M-8 and SHV-28 enzymes in environmental samples, and the present results demonstrate the plasmid-mediated spread of CTX-M-encoding genes through five WWTPs in São Paulo, Brazil, suggesting WWTPs are hotspots for the transfer of ESBL genes and confirming the urgent need to improve the management of sewage in order to minimize the dissemination of resistance genes to the environment.

Characterization of antibiotic resistance in Listeria spp. isolated from slaughterhouse environments, pork and human infections.

INTRODUCTION: Listeria species are susceptible to most antibiotics. However, over the last decade, increasing reports of multidrug-resistant Listeria spp. from various sources have prompted public health concerns. The objective of this study was to characterize the antibiotic susceptibility of Listeria spp. and the genetic mechanisms that confer resistance. METHODOLOGY: Forty-six Listeria spp. isolates were studied, and their minimal inhibitory concentrations of antibiotics were determined by microdilution using Sensititre standard susceptibility MIC plates. The isolates were screened for the presence of gyrA, parC, lde, lsa(A), lnu(A), and mprF by PCR, and the amplified genes were sequenced. RESULTS: All isolates were susceptible to penicillin, ampicillin, tetracycline, erythromycin, and carbapenems. Resistance to clindamycin, daptomycin, and oxacillin was found among L. monocytogenes and L. innocua, and all species possessed at least intermediate resistance to fluoroquinolones. GyrA, parC, and mprF were detected in all isolates; however, mutations were found only in gyrA sequences. A high daptomycin MIC, as reported previously, was observed, suggesting an intrinsic resistance of Listeria spp. to daptomycin. CONCLUSIONS: These results are consistent with reports of emerging resistance in Listeria spp. and emphasize the need for further genotypic characterization of antibiotic resistance in this genus.

A new sterile technique effective on capturing tramp ants for microbiological investigations.

Tramp ants are an outstanding group of organisms that have a definitive and important role in carrying pathogens. The purpose of this study was to develop a microbiological sterile technique of collecting ants from contaminated areas. The traps were composed of Petri dishes containing culture media, and were tested to verify the applicability of the system. A positive correlation between the microorganism growth and the presence of ants inside or around traps was observed. The technique described demonstrated to be useful to collect ants from different environments, helping the surveillance of pathogenic microorganisms that are of public health concern.

Extended-spectrum beta-lactamases among Enterobacteriaceae isolated in a public hospital in Brazil.

Extended-spectrum beta-lactamases (ESBL) in enterobacteria are recognized worldwide as a great hospital problem. In this study, 127 ESBL-producing Enterobacteriaceae isolated in one year from inpatients and outpatients at a public teaching hospital at São Paulo, Brazil, were submitted to analysis by PCR with specific primers for bla SHV, bla TEM and bla CTX-M genes. From the 127 isolates, 96 (75.6%) Klebsiella pneumoniae, 12 (9.3%) Escherichia coli, 8 (6.2%) Morganella morganii, 3 (2.3%) Proteus mirabilis, 2 (1.6%) Klebsiella oxytoca, 2 (1.6%) Providencia rettgeri, 2 (1.6%) Providencia stuartti, 1 (0.8%) Enterobacter aerogenes and 1 (0.8%) Enterobacter cloacae were identified as ESBL producers. Bla SHV, bla TEM and bla CTX-M were detected in 63%, 17.3% and 33.9% strains, respectively. Pulsed field gel eletrophoresis genotyping of K. pneumoniae revealed four main molecular patterns and 29 unrelated profiles. PCR results showed a high variety of ESBL groups among strains, in nine different species. The results suggest the spread of resistance genes among genetically different strains of ESBL-producing K. pneumoniae in some hospital wards, and also that some strongly related strains were identified in different hospital wards, suggesting clonal spread in the institutional environment.

A new sterile technique effective on capturing tramp ants for microbiological investigations / Nova técnica estéril eficiente para captura de formigas andarilhas para investigações microbiológicas

Tramp ants are an outstanding group of organisms that have a definitive and important role in carrying pathogens. The purpose of this study was to develop a microbiological sterile technique of collecting ants from contaminated areas. The traps were composed of Petri dishes containing culture media, and were tested to verify the applicability of the system. A positive correlation between the microorganism growth and the presence of ants inside or around traps was observed. The technique described demonstrated to be useful to collect ants from different environments, helping the surveillance of pathogenic microorganisms that are of public health concern.

Formigas andarilhas são organismos excepcionais, que têm papel definitivo e importante no transporte de patógenos. O objetivo deste estudo foi desenvolver uma técnica microbiologicamente estéril para coletar formigas de áreas contaminadas. As armadilhas compostas por uma placa de Petri contendo meio de cultura foram testadas para verificar a aplicabilidade do sistema. Foi observada correlação positiva entre o crescimento de microrganismos e a presença de formigas dentro ou ao redor das armadilhas. A técnica descrita demonstrou ser útil para coletar formigas de diferentes ambientes, além de contribuir na vigilância de microrganismos patogênicos que representam problema para a saúde pública.

Extended-spectrum beta-lactamases among Enterobacteriaceae isolated in a public hospital in Brazil / Beta-lactamases de espectro estendido em Enterobacteriaceae isoladas de Hospital Público no Brasil

Extended-spectrum β-lactamases (ESBL) in enterobacteria are recognized worldwide as a great hospital problem. In this study, 127 ESBL-producing Enterobacteriaceae isolated in one year from inpatients and outpatients at a public teaching hospital at São Paulo, Brazil, were submitted to analysis by PCR with specific primers for blaSHV, blaTEM and blaCTX-M genes. From the 127 isolates, 96 (75.6 percent) Klebsiella pneumoniae, 12 (9.3 percent) Escherichia coli, 8 (6.2 percent) Morganella morganii, 3 (2.3 percent) Proteus mirabilis, 2 (1.6 percent) Klebsiella oxytoca, 2 (1.6 percent) Providencia rettgeri, 2 (1.6 percent) Providencia stuartti, 1 (0.8 percent) Enterobacter aerogenes and 1 (0.8 percent) Enterobacter cloacae were identified as ESBL producers. BlaSHV, blaTEM and blaCTX-M were detected in 63 percent, 17.3 percent and 33.9 percent strains, respectively. Pulsed field gel eletrophoresis genotyping of K. pneumoniae revealed four main molecular patterns and 29 unrelated profiles. PCR results showed a high variety of ESBL groups among strains, in nine different species. The results suggest the spread of resistance genes among genetically different strains of ESBL-producing K. pneumoniae in some hospital wards, and also that some strongly related strains were identified in different hospital wards, suggesting clonal spread in the institutional environment.

Beta-lactamases de espectro estendido (ESBL) em enterobactérias são reconhecidas mundialmente como um grande problema hospitalar. Neste estudo, 127 Enterobacteriaceae produtoras de ESBL isoladas por um ano, de pacientes internados e ambulatoriais de um hospital público de ensino em São Paulo, Brasil, foram submetidas à análise pela PCR com iniciadores específicos para os genes blaSHV, blaTEM e blaCTX-M. Dos 127 isolados, 96 (75,6 por cento) K. pneumoniae, 12 (9,3 por cento) E. coli, 8 (6,2 por cento) M. morganii, 3 (2,3 por cento) Proteus mirabilis, 2 (1,6 por cento) Klebsiella oxytoca, 2 (1,6 por cento) Providencia rettgeri, 2 (1,6 por cento) Providencia stuartti, 1 (0,8 por cento) Enterobacter aerogenes e 1 (0,8 por cento) Enterobacter cloacae foram identificados como produtores de ESBL. BlaSHV, blaTEM e blaCTX-M foram detectados em 63 por cento, 17,3 por cento e 33,9 por cento das cepas, respectivamente. A genotipagem de K. pneumoniae por eletroforese em campo pulsado revelou quatro padrões moleculares principais e 29 perfis não relacionados. Os resultados da PCR demonstraram alta variedade de grupos de ESBL entre as cepas, em nove espécies diferentes. Os resultados sugerem a disseminação de genes de resistência entre cepas geneticamente diferentes de K. pneumoniae produtoras de ESBL em algumas unidades do hospital, e também que algumas cepas fortemente relacionadas foram identificadas em unidades hospitalares diferentes, sugerindo disseminação clonal no ambiente da instituição.

Characterization of Pseudoalteromonas citrea and P. nigrifaciens isolated from different ecological habitats based on REP-PCR genomic fingerprints.

DNA primers corresponding to conserved repetitive interspersed genomic motifs and PCR were used to show that REP, ERIC and BOX-like DNA sequences are present in marine, oxidative, gram-negative Pseudoalteromonas strains. REP, ERIC and BOX-PCR were used for rapid molecular characterization of both the type species of the genus and environmental strains isolated from samples collected in different geographical areas. PCR-generated genomic fingerprint patterns were found to be both complex and strain specific. Analysis of the genotypic structure of phenotypically diverse P. citrea revealed a geographic clustering of Far Eastern brown-pigmented, agar-digesting strains of this species. Marine isolates of P. nigrifaciens with 67-70% DNA relatedness generated genomic patterns different from those of the type strain and formed a separate cluster. It is concluded that REP, ERIC and BOX-PCR are effective in generating strain specific patterns that can be used to elucidate geographic distribution, with these genomic patterns providing a valuable biogeographic criterion.

Distribution of Potentially Pathogenic Vibrios in Oysters from a Tropical Region.

Oysters ( Crassostrea gigas ) originating from the southern coast of the State of São Paulo-Brazil were analyzed for Vibrio species. Most Probable Numbers (MPN/100 g) were obtained for Vibrio alginolyticus (<3-1,500), Vibrio parahaemolyticus (<3-1,200), Vibrio fluvialis (<3-150), Vibrio cholerae non 01 (<3-40), Vibrio furnissii (<3-40), Vibrio mimicus (<3-40) and Vibrio vulnificus (<3-30). The highest incidence was observed for V. alginolyticus (81%), followed by V. parahaemolyticus (77%), V. cholerae non 01 (31 %), V. fluvialis (27%), V. furnissii (19%), V. mimicus (12%), and V. vulnificus (12%). Forty-eight percent of the isolates tested were positive for enterotoxins in the rabbit ileal loop (RIL) test and 11.1% in the suckling mice test. Vibrio parahaemolyticus (1.1 %) was positive in the Kanagawa test. Vibrio vulnificus (25%) showed lethality in young adult mice. During the field inspection it was observed that generally the conditions of storage were not adequate as 65% of the samples were maintained in temperatures ranging from 25 to 40°C, 19% were frozen, 12% refrigerated and 4% was immersed in contaminated seawater. These results emphasize the great potential for food poisoning by inadequately preserved seafood, and the necessity to upgrade the standards for food quality assessment.