BackgroundKlebsiella pneumoniae is an important opportunistic pathogen associated with nosocomial and community-acquired infections. A wide repertoire of virulence and antimicrobial resistance genes is present in K. pneumoniae genomes, which can constitute extra challenges in the treatment of infections caused by some strains. K. pneumoniae Kp13 is a multidrug-resistant strain responsible for causing a large nosocomial outbreak in a teaching hospital located in Southern Brazil. Kp13 produces K. pneumoniae carbapenemase (KPC-2) but is unrelated to isolates belonging to ST 258 and ST 11, the main clusters associated with the worldwide dissemination of KPC-producing K. pneumoniae. In this report, we perform a genomic comparison between Kp13 and each of the following three K. pneumoniae genomes: MGH 78578, NTUH-K2044 and 342.ResultsWe have completely determined the genome of K. pneumoniae Kp13, which comprises one chromosome (5.3 Mbp) and six plasmids (0.43 Mbp). Several virulence and resistance determinants were identified in strain Kp13. Specifically, we detected genes coding for six beta-lactamases (SHV-12, OXA-9, TEM-1, CTX-M-2, SHV-110 and KPC-2), eight adhesin-related gene clusters, including regions coding for types 1 (fim) and 3 (mrk) fimbrial adhesins. The rmtG plasmidial 16S rRNA methyltransferase gene was also detected, as well as efflux pumps belonging to five different families. Mutations upstream the OmpK35 porin-encoding gene were evidenced, possibly affecting its expression. SNPs analysis relative to the compared strains revealed 141 mutations falling within CDSs related to drug resistance which could also influence the Kp13 lifestyle. Finally, the genetic apparatus for synthesis of the yersiniabactin siderophore was identified within a plasticity region. Chromosomal architectural analysis allowed for the detection of 13 regions of difference in Kp13 relative to the compared strains.ConclusionsOur results indicate that the plasticity occurring at many hierarchical levels (from whole genomic segments to individual nucleotide bases) may play a role on the lifestyle of K. pneumoniae Kp13 and underlie the importance of whole-genome sequencing to study bacterial pathogens. The general chromosomal structure was somewhat conserved among the compared bacteria, and recombination events with consequent gain/loss of genomic segments appears to be driving the evolution of these strains.

Extraintestinal pathogenic Escherichia coli (ExPEC) isolates are responsible for many bloodstream infections. The aim of this study was to characterize E. coli isolated from the bloodstreams of patients ( n = 48) at the University Hospital in Brazil. Epidemiological data were obtained through the analysis of medical records and laboratory tests. By PCR analysis, we investigated the presence of virulence factors (VFs), pathogenicity islands (PAIs), extended-spectrum β-lactamase (ESBL), phylogenetic classifications (A, B1, B2, C, D, E, and F) and molecular genotype by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The mortality analysis showed that 33.3% of the deaths were associated with bacteraemia due to E. coli infections; in addition, an age between 60 and 75 years ( p < 0.001; OR = 6.3[2.1–18.9]) and bacteraemia with an abdominal origin ( p = 0.02; OR = 5[1.2–20.5]) were risk factors for the severity of the infection. Additionally, the presence of the afa gene was associated with mortality due to E. coli bacteraemia ( p = 0.027; OR = 11.4[1.5–85.7]). Immunosuppression (27.1%), intestinal diseases (25.0%) and diabetes (18.8%), were prevalent among patients, and most of the bacteraemia cases were secondary to urinary tract infections (50.0%). The serum resistance gene tra T was present in 77.1% of isolates, group capsular 2 ( kpsMT II) was present in 45.8% and the K5 capsule was present in 20.8% of isolates. The isolates also showed a high prevalence for the siderophore yersiniabactina ( fyu A) (70.8%) and PAI IV 536 (77.1%). Phylogenetic analysis showed that group B2 (45.8%) was the most prevalent, and was the phylogroup that had a higher prevalence of VFs and PAIs. However, in this study, a considerable number of isolated bacteria were classified as group B1 (18.8%) and as group E (14.6%). Eight (16.7%) isolates were resistant to third and fourth generation cephalosporin and group CTX-M-1 (CTX-M-15) was the most prevalent ESBL type. The molecular genotyping showed two clonal lineages and several isolates that were not related to each other. This study provides additional information on the epidemiological and molecular characteristics of E. coli bloodstream infections in Brazil.

New Delhi metallo-β-lactamase 1 (NDM-1) was first identified in Brazil in Enterobacter hormaechei and Providencia rettgeri in 2013. Here, we describe the first case of NDM-1-producing Acinetobacter baumannii sequence type 25 isolated from the urinary tract of a 71-year-old man who died of multiple complications, including A. baumannii infection. The NDM-1 gene was detected by quantitative PCR, and its sequence confirmed its presence in an ∼ 100-kb plasmid.

Microbiological contamination in commercial poultry production has caused concerns for human health because of both the presence of pathogenic microorganisms and the increase in antimicrobial resistance in bacterial strains that can cause treatment failure of human infections. The aim of our study was to analyze the profile of antimicrobial resistance and virulence factors of E. coli isolates from chicken carcasses obtained from different farming systems (conventional and free-range poultry). A total of 156 E. coli strains were isolated and characterized for genes encoding virulence factors described in extraintestinal pathogenic E. coli (ExPEC). Antimicrobial susceptibility testing was performed for 15 antimicrobials, and strains were confirmed as extended spectrum of β-lactamases- (ESBLs-) producing E. coli by phenotypic and genotypic tests. The results indicated that strains from free-range poultry have fewer virulence factors than strains from conventional poultry. Strains from conventionally raised chickens had a higher frequency of antimicrobial resistance for all antibiotics tested and also exhibited genes encoding ESBL and AmpC, unlike free-range poultry isolates, which did not. Group 2 CTX-M and CIT were the most prevalent ESBL and AmpC genes, respectively. The farming systems of poultries can be related with the frequency of virulence factors and resistance to antimicrobials in bacteria.

The frequent use of antimicrobials in commercial poultry production has raised concerns regarding the potential impact of antimicrobials on human health due to selection for resistant bacteria. Several studies have reported similarities between extraintestinal pathogenic Escherichia coli (ExPEC) strains isolated from birds and humans, indicating that these contaminant bacteria in poultry may be linked to human disease. The aim of our study was to analyze the frequency of antimicrobial resistance and virulence factors among E. coli strains isolated from commercial chicken carcasses in Paraná, Brazil, in 2007 and 2013. A total of 84 E. coli strains were isolated from chicken carcasses in 2007, and 121 E. coli strains were isolated in 2013. Polymerase chain reaction was used to detect virulence genes (hlyF, iss, ompT, iron, and iutA) and to determine phylogenetic classification. Antimicrobial susceptibility testing was performed using 15 antimicrobials. The strains were also confirmed as extended-spectrum β-lactamase (ESBL)-producing E. coli with phenotypic and genotypic tests. The results indicated that our strains harbored virulence genes characteristic of ExPEC, with the iutA gene being the most prevalent. The phylogenetic groups D and B1 were the most prevalent among the strains isolated in 2007 and 2013, respectively. There was an increase in the frequency of resistance to a majority of antimicrobials tested. An important finding in this study was the large number of ESBL-producing E. coli strains isolated from chicken carcasses in 2013, primarily for the group 2 cefotaximase (CTX-M) enzyme. ESBL production confers broad-spectrum resistance and is a health risk because ESBL genes are transferable from food-producing animals to humans via poultry meat. These findings suggest that our strains harbored virulence and resistance genes, which are often associated with plasmids that can facilitate their transmission between bacteria derived from different hosts, suggesting zoonotic risks.

Infecções associadas aos cuidados de saúde (IRAS) constituem importante preocupação à segurança do paciente ao redor do mundo. Pneumonia associada à ventilação mecânica (VAP) é a principal causa de morte entre IRAS, com mortalidade de 15 a 70%, dependendo da população de pacientes. O Centro de Controle e Prevenção de Doenças (CDC) recomendou que todas as UTIs implementassem um bundle de para reduzir a taxa de PAV. Os objetivos do presente estudo foram analisar o efeito do pacote de intervenções de controle da infecção, a educação, as taxas de PAV na Unidade Pediátrica de Terapia Intensiva (UTIP) do Hospital Universitário de Londrina-PR. Este estudo foi realizado entre janeiro e dezembro de 2013 e consistiu em três períodos: pré-intervenção, intervenção e pós-intervenção. A intervenção educativa foi dada a 86 trabalhadores da saúde (PS), e um questionário também foi realizado no pré e pós-intervenção. No geral, foram avaliadas 135 oportunidades de atendimento ao paciente. A higiene das mãos diminuiu do período pré para o pós-intervenção, entretanto a intubação gástrica por via oral, a manutenção da cabeceira da cama entre 30-45º, a pressão do cuff endotraqueal e remoção de condensação do circuito respirador aumentaram significativamente do período pré para o pós-intervenção. A taxa de PAV foi 49,6% durante o período de pré-intervenção e 17,5% durante o período pós-intervenção demonstrando uma redução de 64,8%. Nossos resultados mostram que a implementação do pacote de intervenções de controlo da infecção foi associada com uma redução significativa na taxa de VAP. ResumoPalavras-chave: Pneumonia associada à ventilação. Bundle. Prevenção. PAV. AbstractHealthcare-associated infections (HAI) are an important patient safety concern around the globe. Ventilatorassociated pneumonia (VAP) is the leading cause of death among HAI, with attributable mortality ranging from 15 to 70% depending on the patient population. The Center for Disease Control and Prevention (CDC)

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Introduction: Extraintestinal pathogenic Escherichia coli (ExPEC) is associated with various diseases such as urinary tract infections, neonatal meningitis and septicemia. There are many virulence factors (VF) encoded by genes in ExPEC, including papC, papG, ecpA, iroN, fyuA, iutA, ompTp, tsh, hlyF, hlyA and iss. These virulence genes may be present in pathogenicity islands (PAI) or plasmids. Methodology: In this study, we analyzed the presence of VF encoding genes, PAI sequences and phylogenetic groups of 96 ExPEC strains isolated from the urine and blood of patients at the University Hospital of Londrina, and we compared them with 50 faecal commensal strains from healthy individuals. Results: The VF fyuA (65.60%) was detected in pathogenic strains and commensal strains (46%). A comparison of the distribution of ExPEC and commensal strains in the phylogenetic groups showed that more ExPEC strains belonged to group B2 whereas more of the commensal isolates belonged to group A. The distribution of the seven PAI sequences between commensal strains and ExPEC strains showed that PAI IV 536 was common in both ExPEC and commensal isolates. Conclusions: These results showed that the ExPEC strains that belonged to group B2 had more PAI sequences compared to those of the other groups, especially group B1, which had virulence genes but the lowest percentage of PAI sequences, which leads us to conclude that the virulence of ExPEC strains characterized as B2 is likely attributed to PAI encoded genes, whereas the virulence of ExPEC strains belonging to phylogenetic group B1 is likely due to plasmid encoded virulence genes.