Gut colonization and subsequent infection of neonates caused by extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae

The gut microbiota harbors diverse bacteria considered reservoirs for antimicrobial resistance genes. The global emergence of extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales (ESBL-PE) significantly contributes to healthcare-associated infections (HAIs). We investigated the presence of ESBL-producing Escherichia coli (ESBL-PEco) and ESBL-producing Klebsiella pneumoniae (ESBL-PKpn) in neonatal patients’ guts. Furthermore, we identified the factors contributing to the transition towards ESBL-PEco and ESBL-PKpn-associated healthcare-associated infections (HAIs). The study was conducted from August 2019 to February 2020, in a Neonatal Intensive Care Unit of the Hospital Infantil de México Federico Gómez. Rectal samples were obtained upon admission, on a weekly basis for a month, and then biweekly until discharge from the neonatology ward. Clinical data, culture results, and infection information were gathered. We conducted antimicrobial tests, multiplex PCR assay, and pulsed-field gel electrophoresis (PFGE) to determine the antimicrobial resistance profile and genetic relationships. A comparison between the group’s controls and cases was performed using the Wilcoxon and Student t-tests. Of the 61 patients enrolled, 47 were included, and 203 rectal samples were collected, identifying 242 isolates. In 41/47 (87%) patients, colonization was due to ESBL-PEco or ESBL-PKpn. And nine of them developed HAIs (22%, 9/41). ESBL-PEco resistance to cephalosporins ranged from 25.4% to 100%, while ESBL-PKpn resistance varied from 3% to 99%, and both bacteria were susceptible to carbapenems, tigecillin, and colistin. The prevalent blaCTX-M-group-1 gene accounted for 77.2% in ESBL-PEco and 82.2% in ESBL-PKpn, followed by blaTEM 50% and blaOXA-1 43.8% in ESBL-PEco and blaTEM 80.2% and blaSHV 76.2% in ESBL-PKpn. Analysis of clonality revealed identical colonizing and infection isolates in only seven patients. Significant risk factors included hospital stay duration, duration of antibiotic treatment, and invasive device usage. Our findings suggest high ESBL-PEco and ESBL-PKpn rates of colonization often lead to infection in neonates. Attention should be paid to patients with ESBL-PE

Pandrug-resistant Acinetobacter baumannii from different clones and regions in Mexico have a similar plasmid carrying the blaOXA-72 gene

BackgroundMultidrug-resistant Acinetobacter baumannii is a common hospital-acquired pathogen. The increase in antibiotic resistance is commonly due to the acquisition of mobile genetic elements carrying antibiotic resistance genes. To comprehend this, we analyzed the resistome and virulome of Mexican A. baumannii multidrug-resistant isolates.MethodsSix clinical strains of A. baumannii from three Mexican hospitals were sequenced using the Illumina platform, the genomes were assembled with SPAdes and annotated with Prokka. Plasmid SPAdes and MobRecon were used to identify the potential plasmid sequences. Sequence Type (ST) assignation under the MLST Oxford scheme was performed using the PubMLST database. Homologous gene search for known virulent factors was performed using the virulence factor database VFDB and an in silico prediction of the resistome was conducted via the ResFinder databases.ResultsThe six strains studied belong to different STs and clonal complexes (CC): two strains were ST208 and one was ST369; these two STs belong to the same lineage CC92, which is part of the international clone (IC) 2. Another two strains were ST758 and one was ST1054, both STs belonging to the same lineage CC636, which is within IC5. The resistome analysis of the six strains identified between 7 to 14 antibiotic resistance genes to different families of drugs, including beta-lactams, aminoglycosides, fluoroquinolones and carbapenems. We detected between 1 to 4 plasmids per strain with sizes from 1,800 bp to 111,044 bp. Two strains from hospitals in Mexico City and Guadalajara had a plasmid each of 10,012 bp pAba78r and pAba79f, respectively, which contained the blaOXA-72 gene. The structure of this plasmid showed the same 13 genes in both strains, but 4 of them were inverted in one of the strains. Finally, the six strains contain 49 identical virulence genes related to immune response evasion, quorum-sensing, and secretion systems, among others.ConclusionResistance to carbapenems due to pAba78r and pAba79f plasmids in Aba pandrug-resistant strains from different geographic areas of Mexico and different clones was detected. Our results provide further evidence that plasmids are highly relevant for the horizontal transfer of antibiotic resistance genes between different clones of A. baumannii

Sulfhydryl variable-5 extended spectrum β-lactamase in nosocomial enteric bacteria causing sepsis in mexican children

Introduction: Enteric bacteria causing nosocomial infections are often resistant to third-generation cephalosporins due to the production of extended-spectrum β-lactamases (ESBLs). Objective: To describe and characterize the ESBLs pattern present in Klebsiella pneumoniae and Serratia marcescens strains, isolated as causative of nosocomial sepsis in pediatric patients at Instituto Nacional de Pediatría (National Institute of Pediatrics). Material and methods: We analyzed 94 strains of K. pneumoniae and 7 of S. marcescens isolated from clinical specimens from 2002-2005, causative of sepsis in a children’s hospital. We evaluated antibiotic susceptibility and detection of ESBL phenotypes by disk diffusion methods; ceftazidime-resistant isolates were further characterized by pulsed field gel electrophoresis (PFGE); and ESBLs were phenotypically and genotypically characterized by isoelectric focusing, polymerase chain reaction (PCR) and sequencing. We also assed for presence of conjugative plasmids bearing the ESBL gene. Results: 51/94 (54%) of K. pneumoniae isolates, and 5/7 (71%) of S. marcescens isolates were resistant to ceftazidime; all carried a blaSHV-5 gene. All K. pneumoniae isolates had a distinct PFGE profile, yet all carried a ~48-Kb plasmid, that was conjugatively transferable to an Escherichia coli receptor, which expressed the resistance phenotype. On the other hand, all S. marcescens isolates had a similar PFGE profile, were unable to transfer the ceftazidime-resistance phenotype, and were isolated from the same ward in a short time-span suggesting an outbreak. Conclusions: The overall prevalence of ESBL-producing enteric bacteria in this hospital is high but similar to other Latin American reports. The sulfhydryl variable-5 (SHV-5) ESBL gene appears to reside in a highly mobile plasmid, capable of spreading among different K. pneumoniae clones and perhaps even to S. marcescens

Association of Antibiotic Resistance, Cell Adherence, and Biofilm Production with the Endemicity of Nosocomial Klebsiella pneumoniae

Klebsiella pneumoniae is a leading cause of multiple nosocomial infections, some of which are associated with high mortality. The increasing prevalence of antibiotic-resistant strains highlights their clinical importance and how complicated managing treatment can be. In this study, we investigated antimicrobial resistance, cell adherence, and biofilm production of nosocomial K. pneumoniae strains isolated from surveillance studies in a Mexican tertiary hospital and evaluated the potential association of these phenotypes with endemicity. The great majority of the clones exhibited adhesion to cultured epithelial cells and were strong biofilm producers. A direct relationship between adhesion phenotypes, biofilm production, and endemicity was not always apparent. Biofilm formation and production of ESBL did not appear to be directly associated. Notably, all the endemic strains were multidrug-resistant. This study emphasizes that while endemic strains possess various virulence-associated properties, antimicrobial resistance appears to be a determining factor of their endemicity

Risk factors for extended-spectrum b-lactamases-producing Escherichia coli urinary tract infections in a tertiary hospital

Objective. To assess the risks factors for urinary tract infections (UTIs) caused by Extended-Spectrum Beta-Lactamases (ESBLs)-producing E. coli and the molecular characterization of ESBLs. Materials and methods. A case-control study was performed to identify risk factors in consecutively recruited patients with UTIs caused by ESBLs or non-ESBLs-producing E. coli in a tertiary hospital in Mexico. Results. ESBLs-producing E. coli were isolated from 22/70 (31%) patients with E. coli UTIs over a three month period. All isolates were resistant to cephalosporins and quinolones but susceptible to carbapenems, amikacin and nitrofurantoin. Prior antibiotic treatment with more than two antibiotic families (OR=6.86; 95%CI 1.06-157.70; p=0.028), recurrent symptomatic UTIs (OR=5.60; 95%CI 1.88-17.87; p=0.001) and previous hospitalization (OR=5.06; 95%CI 1.64-17.69;p=0.002) were significant risk factors. Sixteen isolates harbored the beta-lactamase (bla)CTX-M-15 gene and five the blaTEM-1 gene. Conclusions. One of every three patients presented UTIs with ESBLs-producing beta-lactams and fluoroquinolone resistant E. coli. Risk factors and resistance patterns must be taken into account for developing antibiotic use policies in these setting

Antibiotic susceptibility for 112 nosocomial isolates of <i>Acinetobacter baumannii</i>.

<p>Antibiotic susceptibility for 112 nosocomial isolates of <i>Acinetobacter baumannii</i>.</p

Frequency analysis of <i>A</i>. <i>baumannii</i> clones during 2014.

<p>(A) Frequency of each clone isolated by ward. (B) Frequency of each clone by month of isolation.</p

Analysis of serum resistance activity by <i>A</i>. <i>baumannii</i> clones.

<p>Each <i>A</i>. <i>baumannii</i> isolate was assessed by its ability to survive in normal human serum (NHS). (A) We show the percentage of <i>A</i>. <i>baumannii</i> isolates to survive in presence of 40% of NHS. (B) We show the ability of the members of each clone to survive at 40% of NHS. The dotted lines indicate the survival rate (0–25, 26–50, 51–75 and 76–100%) in presence of 40% of NHS. Open circle indicate patients that improved and every closed circle corresponded to a patient that died. Each point corresponds to the average of two independent experiments by duplicate. Each column indicates the SD.</p

<i>Acinetobacter baumannii</i> clones and mechanisms associated with carbapenem resistance.

<p><i>Acinetobacter baumannii</i> clones and mechanisms associated with carbapenem resistance.</p