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Introduction Proteus mirabilis is one of the main pathogens that cause urinary tract infections. Therefore, the aim of this study was to analyze and compare the genetic profile of 36 clinical isolates of P. mirabilis that carry and do not carry the blaKPC and blaNDM gene with respect to virulence factors (mrpG, pmfA, ucaA, nrpG and pbtA) and antimicrobial resistance (blaVIM, blaIMP, blaSPM, blaGES, blaOXA-23-like, blaOXA-48-like, blaOXA-58-like and blaOXA-10-like). Methods The virulence and resistance genes were investigated by using PCR and sequencing. Results ERIC-PCR typing showed that the isolates showed multiclonal dissemination and high genetic variability. The gene that was most found blaOXA-10-like (n = 18), followed by blaKPC (n = 10) and blaNDM (n = 8). To our knowledge, this is the first report of blaOXA-10 in P. mirabilis in Brazil, as well as the first report of the occurrence of P. mirabilis co-carrying blaOXA-10/blaKPC and blaOXA-10/blaNDM. The blaNDM or blaKPC carrier isolates showed important virulence genes, such as ucaA (n = 8/44.4%), pbtA (n = 10/55.5%) and nrpG (n = 2/11.1%). However, in general, the non-carrier isolates of blaKPC and blaNDM showed a greater number of virulence genes when compared to the carrier group. Conclusion Clinical isolates of P. mirabilis, in addition to being multi-drug resistant, presented efficient virulence factors that can establish infection outside the gastrointestinal tract.

Objectives Multidrug-resistant Klebsiella pneumoniae carrying blaNDM-1 and blaKPC-2 genes are a worldwide concern for which combination antimicrobial therapy may be the only viable option. The aim of this study was to investigate the in vitro activity of combinations of polymyxin B (PMB) with meropenem (MEM), amikacin (AMK) and gentamicin (GEN) at subinhibitory concentrations against two K. pneumoniae clinical isolates co-harbouring blaNDM-1, blaKPC-2 and aminoglycoside-modifying enzymes and resistant to PMB. Methods Synergy and bactericidal activity were evaluated by chequerboard and time–kill assays against two PMB-resistantK. pneumoniae clinical isolates carrying the blaNDM-1, blaKPC-2, aac(3)-IIa, aac(6ʹ)-Ib, aph(3ʹ)-VI and ant(2ʹʹ)-Ia genes. Five combinations of PMB, MEM, AMK and GEN were evaluated. Results The PMB/MEM and PMB/AMK combinations proved to be the best options against isolate K7R2, mainly because they demonstrated bactericidal activity when using subinhibitory concentrations of these antimicrobials. However, none of the studied combinations was bactericidal against isolate K11R2. Conclusion The combinations used in this study showed synergy against NDM-and KPC-producing isolates but, given their bactericidal activity, the combinations of PMB/MEM and PMB/AMK were the most active against one isolate. It can also be concluded that the antimicrobials to which the bacteria were resistant could form part of combination therapy.

Objectives Carbapenemase-producing Enterobacterales are frequently involved in healthcare-associated infections worldwide. The objectives of this study were to investigate (i) the frequency of the main genes encoding carbapenemases, 16S rRNA methylases and aminoglycoside-modifying enzymes (AMEs) as well as the mcr gene and (ii) the clonal relationship of enterobacteria isolates resistant to carbapenems and aminoglycosides from colonisation and infection in patients from hospitals in northeastern Brazil. Methods Antimicrobial susceptibility was determined using an automated VITEK®2 system. Presence of carbapenemase, AME and 16S rRNA methylase genes as well as the mcr gene was determined by PCR and amplicon sequencing. Genetic variability was determined by ERIC-PCR. Results A total of 35 isolates resistant to carbapenems and aminoglycosides were selected for this study. Klebsiella pneumoniae was most common (45.7%), followed by Proteus mirabilis (28.6%) and Serratia marcescens (25.7%). AME genes were found in 97.1% of isolates, most commonly aph(3ʹ)-VI and aac(6')-Ib. The blaNDM-1 and blaKPC-2 genes were detected in 25.7% and 88.6% of isolates, respectively; five isolates harboured these genes concomitantly. According to the literature, this is the first report of the association of blaNDM-1 and blaKPC-2 in P. mirabilis and S. marcescens in Brazil. The isolates showed a multiclonal profile by ERIC-PCR. Conclusion The emergence of blaNDM-1 associated with blaKPC-2 and AME genes in K. pneumoniae, P. mirabilis and S. marcescens isolates with a multiclonal profile is of concern as this limits therapeutic options. These results should alert medical authorities to establish rigorous detection methods to reduce the spread of these antimicrobial resistance genes.

INTRODUCTION: Carbapenemase-resistant enterobacteria that produce the bla NDM gene are found worldwide. However, this is the first report of blaNDM in Klebsiella aerogenes in Brazil. METHODS: The identification of bacterial species was performed using anautomated system and confirmed by biochemical tests, 16S rRNA gene sequencing, and detection of resistance genes. RESULTS: The clinical isolate showed minimum inhibitory concentration resistance to meropenem and polymyxin B at 8mg/L and 4mg/L, respectively. Only the blaNDM gene was detected. CONCLUSIONS: The current report of the blaNDM gene in isolated MDR enterobacteria indicates that this gene can spread silently in a hospital setting.

INTRODUCTION: This study investigated the genetic environment of bla KPC-2 in Klebsiella pnemoniae multi-drug resistant clinical isolates. METHODS: Four carbapenemase gene isolates resistant to carbapenems, collected from infected patients from two hospitals in Brazil, were investigated using polymerase chain reaction and plasmid DNA sequencing. RESULTS: The bla KPC-2 gene was located between ISKpn6 and a resolvase tnpR in the non-Tn4401 element (NTEKPC-IId). It was detected on a plasmid belonging to the IncQ1 group. CONCLUSIONS To our knowledge, this is the first report of the presence of the bla KPC-2 gene in the NTEKPC-IId element carried by plasmid IncQ1 from infections in Brazil.

Purpose: To analyze the presence of microorganisms in fluorescein eyedrops used in a reference eye center in Recife-PE. Methods: This real-life and masked study evaluated fluorescein eyedrops used at the Altino Ventura Foundation in May 2019. Cultures were performed according to exposure times; I) three eyedrop bottles were analyzed after one day of use, II) three eyedrop bottles after 4 d of use, III) three eyedrop bottles after 8 d of use, and IV) three unopened bottles used as control. Samples were collected from the bottle’s tip, instilled drop, and residual fluid. After incubation, all colonies were analyzed and identified through biochemical tests. Results: The contamination rate of the fluorescein eyedrop bottles in this study was 55.5% (5/9 vials). There was no contamination in the control group. The highest contamination was seen in one day exposed eyedrops, in 100% of the bottles. The bottle’s tip had a higher rate of contamination compared to the drop and residual fluid. Gram-positive bacteria were isolated in 7/27 (25.9%) samples. Growth of fungi or gram-negative bacteria was not observed. Conclusion: The identification of gram-positive bacteria predominantly on the tip of the fluorescein eyedrop bottles suggests inadequate handling as the main cause of contamination. RESUMO Objetivo: Analisar a presença de microrganismos nos colírios de fluoresceína utilizados em um centro oftalmológico de referência em Recife-PE. Métodos: Este estudo de vida real e mascarado avaliou colírios de fluoresceína utilizados na Fundação Altino Ventura em maio/2019. As culturas foram realizadas de acordo com os diferentes tempos de exposição: I - três frascos de colírio foram analisados após 1 dia de uso; II - três frascos de colírio após 4 dias de uso; III - três frascos de colírio após 8 dias de uso; IV - três garrafas fechadas foram usadas como grupo controle. As amostras foram coletadas da ponta do frasco, da gota instilada e do líquido residual interior. Após incubação, todas as colônias foram analisadas e identificadas através de testes bioquímicos. Resultados: A taxa de contaminação dos frascos de colírio de fluoresceína neste estudo foi de 55,5% (5/9 frascos). Não houve contaminação no grupo controle. A maior contaminação foi observada os colírios expostos de um dia - 100% dos frascos. A ponta da garrafa teve uma maior taxa de contaminação em comparação com as culturas de gota e de fluido residual inferior. Bactérias gram-positivas foram isoladas em 7/27 amostras (25,9%). Não houve crescimento de fungos ou bactérias Gram-negativas. Conclusão: A identificação de bactérias Gram-positivas predominantemente na ponta dos frascos de colírio de fluoresceína sugere manuseio inadequado como a principal causa de contaminação de colírios multidose.

A resistência a carbapenemases em bacilos gram-negativas (BGN) tornou-se um problema mundial, elevando a taxa de morbidade e mortalidade. O método automatizado foi utilizado para identificação e suscetibilidade dos isolados. E o método molecular foi aplicado na detecção de bacilos gram-negativas (BGN) produtoras de carbapenemase. Um total de 205 BGN foram recuperados a partir de diferentes amostras clínicas, resistentes a carbapenemases. Os genes de resistência blaKPC (57,5%), blaVIM (30,2%), blaGES (17%), blaNDM (15%) e blaSPM (2,4%) foram recuperados a partir de isolados clínicos. O gene blaIMP não foi detectado em nenhum dos isolados. É preocupante que Pseudomonas aeruginosa e Acinetobacter baumannii representem a maioria dos isolados (61,6%) recuperados de infecções de diferentes sítios, com alta resistência a carbapenêmicos, carregando simultaneamente os genes blaKPC, blaGES e blaVIM, assim como encontrados também em Enterobactericeae. A carbapenemase mais prevalente foi blaKPC e blaVIM, seguida de blaNDM dentre os isolados multidroga resistentes. Esses resultados são um ameaça à saúde pública, configurando altas taxas de resistência, limitando as opções terapêuticas.

INTRODUCTION: The emergence of New Delhi metallo-β-lactamase (NDM) is concernig because it reduces the antibiotic therapy options for bacterial infections. METHODS: Resistant and virulent genes from an isolate of Klebsiella pneumoniae derived from a patient with sepsis in a hospital in Recife-PE, Brazil, were investigated using PCR and DNA sequencing. RESULTS: bla NDM-1, aac(6’)-Ib-cr and acrB resistance genes, and cps and mrkD virulence genes were detected. CONCLUSIONS To our knowledge, this is the first report on bla NDM-1 in Recife-PE. This detection alerts researchers to the need to control the spread of bla NDM-1 resistance gene by this bacterium in Brazil.

INTRODUCTION: The objective of this study was to characterize genes of aminoglycoside modifying enzymes (AMEs) in colonizing and infecting isolates of E. aerogenes harboring bla KPC from patients at a public hospital in Recife-PE, Brazil. METHODS: We analyzed 29 E. aerogenes clinical isolates resistant to aminoglycosides. AMEs genes were investigated by PCR and sequencing. RESULTS: Colonizing and infecting isolates mainly presented the genetic profiles aac(3)-IIa/aph(3’)-VI or ant(2”)-IIa/aph(3’)-VI. This is the first report of aph(3’)-VI in E. aerogenes harboring bla KPC in Brazil. CONCLUSIONS: The results highlight the importance in establishing rigorous methods for the surveillance of resistance genes, especially in colonized patients.