Virulence genes from different E. coli pathotypes are blended in hybrid strains. E. coli strains with hybrid enteroaggregative/uropathogenic (EAEC/UPEC) genotypes have sporadically emerged causing outbreaks of extraintestinal infections, however their association with routine infections is yet underappreciated. We assessed 258 isolates of E. coli recovered from 86 consecutive cases of extraintestinal infections seeking EAEC and hybrid genotype (EAEC/UPEC) strains. Extensive virulence genotyping was carried out to detect 21 virulence genes, including molecular predictors of EAEC and UPEC strains. Phylogenetic groups and sequence types (STs) were identified, as well as it was performed phylogenetic analyses in order to evaluate whether hybrid EAEC/UPEC strains belonged to intestinal or extraintestinal lineages of E. coli. Adhesion assays were performed to evaluate the biofilm formation by hybrid strains in human urine and cell culture medium (DMEM). Molecular predictors of UPEC were detected in more than 70% of the strains (chuA in 85% and fyuA in 78%). Otherwise, molecular predictors of EAEC (aatA and aggR) were detected in only 3.4% (9/258) of the strains and always along with the UPEC predictor fyuA. Additionally, the pyelonephritis-associated pilus (pap) gene was also detected in all of the hybrid EAEC/UPEC strains. EAEC/UPEC strains were recovered from two cases of community-onset urinary tract infections (UTI) and from a case of bacteremia. Analyses revealed that hybrid EAEC/UPEC strains were phylogenetically positioned in two different clades. Two representative strains, each recovered from UTI and bacteremia, were positioned into a characteristic UPEC clade marked by strains belonging to phylogenetic group D and ST3 (Warwick ST 69). Another hybrid EAEC/UPEC strain was classified as phylogroup A-ST478 and positioned in a commensal clade. Hybrid EAEC/UPEC strains formed biofilms at modest, but perceptible levels either in DMEM or in urine samples. We showed that different lineages of E. coli, at least phylogenetic group A and D, can acquire and gather EAEC and UPEC virulence genes promoting the emergence of hybrid EAEC/UPEC strains.
Background
Aquatic matrices impacted by sewage may shelter carbapenem-resistant (CR) Gram-negative bacilli (GNB) harboring resistance genes of public health concern. In this study, sewage treatment plants (STPs) servicing well-defined catchment areas were surveyed for the presence of CR-GNB bearing carbapenemase genes (blaKPC or blaNDM).
Results
A total of 325 CR-GNB were recovered from raw (RS) and treated (TS) sewage samples as well as from water body spots upstream (UW) and downstream (DW) from STPs. Klebsiella-Enterobacter (KE) group amounted to 116 isolates (35.7%). CR-KE isolates were recovered from TS, DW (35.7%) and RS samples (44.2%) (p = 0.001); but not from UW samples. KE isolates represented 65.8% of all blaKPC or blaNDM positive strains. The frequency of blaKPC-or-NDM strains was positively associated with the occurrence of district hospitals located near STPs, as well as with the number of hospitalizations and of sewer connections serviced by the STPs. blaKPC-or-NDM strains were recovered from ST samples in 7 out of 14 STPs, including four tertiary-level STPs; and from 6 out of 13 DW spots whose RS samples also had blaKPC-or-NDM strains.
Conclusions
Clinically relevant GNB bearing blaKPC-or-NDM resist sewage treatments and spread into environmental aquatic matrices mainly from STPs impacted by hospital activities.
Human enteric viruses, such as enteric adenoviruses (HAdV), are known to be involved with gastrointestinal disorders, especially acute gastroenteritis. Several studies have used HAdV as an indicator of water quality, since they are considered highly stable and widely distributed viruses in water matrices. The aim of this study was to detect and genotype HAdVs in water matrices impacted by discharges of treated effluents from wastewater treatment plants (WWTPs). Wastewater treatment plants from the sanitary system of the Brazilian Federal District were assessed in 2018 and 2019. Samples were collected upstream and downstream from discharge points for each WWTP. Viral concentration based on adsorption-elution and conventional PCR was used for molecular detection, and positive samples were sequenced for phylogenetic analysis. Pluviosity data for the period in which the samples were collected were obtained. Our results demonstrated the presence of HAdVs in 27.2% (61/224) of the samples. The positivity was significantly higher in downstream samples compared to upstream. Moreover, the HAdV positivity was higher in downstream samples collected from receiving water bodies impacted by secondary-level WWTPs in comparison with those impacted by tertiary-level WWTPs. Phylogenetic analysis demonstrated the presence of genotypes 40 and 41, with prevalence of HAdV genotype 41. Despite the predominance of HAdV-41, an increasing frequency of the HAdV-40 was associated with higher pluviosity. In conclusion, this study is the first documentation in the Brazilian Federal District dealing with the prevalence and diversity of HAdVs in several WWTP, along with their correlation with rainfall index.
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