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Article

Characterization of Uropathogenic Escherichia coli Reveals Hybrid Isolates of Uropathogenic and Diarrheagenic (UPEC/DEC) E. coli

by
Rodrigo H. S. Tanabe
1,
Regiane C. B. Dias
1,
Henrique Orsi
1,
Daiany R. P. de Lira
1,
Melissa A. Vieira
1,
Luís F. dos Santos
2,
Adriano M. Ferreira
3,
Vera L. M. Rall
1,
Alessandro L. Mondelli
4,
Tânia A. T. Gomes
5,
Carlos H. Camargo
2 and
Rodrigo T. Hernandes
1,*
1
Departamento de Ciências Químicas e Biológicas (Setor de Microbiologia e Imunologia), Instituto de Biociências, Universidade Estadual Paulista (UNESP), Botucatu 18618-689, SP, Brazil
2
Centro de Bacteriologia, Instituto Adolfo Lutz, São Paulo 01246-902, SP, Brazil
3
Hospital das Clínicas da Faculdade de Medicina de Botucatu, Botucatu 18607-741, SP, Brazil
4
Departamento de Clínica Médica, Faculdade de Medicina, Universidade Estadual Paulista (UNESP), Botucatu 18618-970, SP, Brazil
5
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo-Escola Paulista de Medicina (UNIFESP-EPM), São Paulo 04023-062, SP, Brazil
*
Author to whom correspondence should be addressed.
Microorganisms 2022, 10(3), 645; https://doi.org/10.3390/microorganisms10030645
Submission received: 8 February 2022 / Revised: 8 March 2022 / Accepted: 15 March 2022 / Published: 17 March 2022
(This article belongs to the Section Antimicrobial Agents and Resistance)
Versions Notes

Abstract

:
(1) Background: Pathogenic Escherichia coli are divided into two groups: diarrheagenic (DEC) and extraintestinal pathogenic (ExPEC) E. coli. ExPEC causing urinary tract infections (UTIs) are termed uropathogenic E. coli (UPEC) and are the most common cause of UTIs worldwide. (2) Methods: Here, we characterized 112 UPEC in terms of phylogroup, serotype, the presence of virulence factor-encoding genes, and antimicrobial resistance. (3) Results: The majority of the isolates were assigned into the phylogroup B2 (41.07%), and the serogroups O6 (12.5%) and O25 (8.9%) were the most frequent. Five hybrid UPEC (4.5%), with markers from two DEC pathotypes, i.e., atypical enteropathogenic (aEPEC) and enteroaggregative (EAEC) E. coli, were identified, and designated UPEC/aEPEC (one isolate) and UPEC/EAEC (four isolates), respectively. Three UPEC/EAEC harbored genes from the pap operon, and the UPEC/aEPEC carried ibeA. The highest resistance rates were observed for ampicillin (46.4%) and trimethoprim/sulfamethoxazole (34.8%), while 99.1% of the isolates were susceptible to nitrofurantoin and/or fosfomycin. Moreover, 9.8% of the isolates were identified as Extended Spectrum β-Lactamase producers, including one hybrid UPEC/EAEC. (4) Conclusion: Our data reinforce that hybrid UPEC/DEC are circulating in the city of Botucatu, Brazil, as uropathogens. However, how and whether these combinations of genes influence their pathogenicity is a question that remains to be elucidated.

1. Introduction

Escherichia coli, a member of the family Enterobacteriaceae, is generally a harmless commensal of the gastrointestinal tract [1]. However, there are several E. coli clones that have acquired the ability to produce virulence factors, which provides them with the potential to cause several infectious diseases in humans and animals [2,3]. Genes encoding virulence factors associated with the E. coli pathogenesis are often located in mobile genetic elements, such as plasmid, phage, and pathogenic island (PAI), that can be mobilized into different E. coli backgrounds, thus originating many distinct genetic profiles [4,5,6,7,8,9].
Pathogenic E. coli are divided into two major groups: diarrheagenic (DEC) and extraintestinal pathogenic E. coli (ExPEC), which causes diarrhea and extraintestinal infections, such as neonatal meningitis, sepsis, and urinary tract infections (UTIs), respectively [2,3,10]. Based mainly on the virulence repertory and phenotypic features, DEC is currently classified into six distinct pathotypes [2]. Atypical enteropathogenic (aEPEC), which differs from typical EPEC (tEPEC) by the absence of the EPEC adherence factor (EAF) plasmid [11], and enteroaggregative (EAEC) E. coli are the most common DEC pathotypes isolated from diarrheal patients in Brazil [12,13] as well as worldwide [14], even though aEPEC and EAEC have also been frequently isolated from healthy subjects with asymptomatic intestinal infections [13,15,16,17]. In addition, E. coli isolates harboring EPEC and EAEC markers have also been implicated as etiologic agents of extraintestinal infections, i.e., UTIs and bloodstream infections [18,19,20,21,22].
E. coli isolates causing UTIs are collectively known as uropathogenic E. coli (UPEC) and are the most common cause of this infection in both out- and inpatients worldwide [23]. Virulence strategies of UPEC include adherence to urinary tract epithelial cells, iron acquisition, toxins production, and evasion of the host’s immune system [24,25]. These virulence factors are frequently encoded by genes located in pathogenicity islands (PAIs), most often acquired by horizontal gene transfer [6,26,27].
Phylogenomic analyses have consistently demonstrated that the E. coli species is very complex and structured in eight distinct phylogroups, as follows: A, B1, B2, C, D, E, F, and the newly described G [28,29,30]. The large majority of the UPEC isolates have been assigned into phylogroup B2, in addition to isolates observed in other E. coli phylogroups [31,32].
Mainly since the mid-2000s, the constant increase of UPEC rates with resistance to several antimicrobial drugs has been undermining the effectiveness of clinical management of UTIs, thus becoming a worldwide public health concern [33]. Of particular importance we can highlight the UPEC isolates that produce extended-spectrum beta-lactamases (ESBL) enzymes and/or are multidrug-resistant (MDR) [34,35].
The main goal of this study was to characterize a collection of UPEC isolates obtained from outpatients of the Hospital of the Medical School of Botucatu, Brazil, regarding their serotype, presence of virulence factor-encoding genes, and antimicrobial resistance profiles. In addition, all the isolates were assigned in the distinct E. coli phylogroups.

2. Material and Methods

2.1. UPEC Isolates

This study analyzed 112 UPEC isolates obtained from outpatient urine samples with counts ≥105 colony-forming units (CFU) per mL of urine. The 112 outpatients were attended to at the Clinical Hospital from the Medical School–University of São Paulo State (UNESP)–Botucatu, São Paulo State, Brazil, between March, April, and May 2018.

2.2. Virulence Factor-Encoding Genes Detection

The presence of a total of 29 genes encoding proteins associated with distinct roles in the pathogenesis of ExPEC isolates, such as adhesins (fimH, ecpA, papA, papC, iha, afaBC, sfa/focDE, hra, and bamE), toxins (usp, sat, vat, hlyA, hlyF, picU, cnf1, tsh, and cdt), iron acquition (iroN, irp2, iucD, ireA, and sitA), serum resistance (tratT, ompT, iss, cva, and kpsMTII) and the invasion-encoding gene ibeA, were investigated in the UPEC isolates by DNA amplification. Polymerase Chain Reaction (PCR) was performed using Green Master Mix (Promega, Madison, WI, USA) with 0.34 μM of each primer. All primer sequences and PCR conditions used to detect virulence genes are described in the references cited in Table S1.
Moreover, genes that are frequently used for the diagnosis of some DEC pathotypes, such as EPEC (eae and bfpB), EAEC (aggR and aatA), and STEC (stx1/2), were investigated using primers described in Table S2. Positive and negative controls were included in all PCR reactions as described in previous publications from our laboratory [17,36].

2.3. Serotyping

The identification of somatic (O) and flagellar (H) antigens was performed by standard agglutination tests [37], with specific O1–O181 and H1–H56 antisera produced at the Instituto Adolfo Lutz, São Paulo, Brazil [12].

2.4. E. coli Phylogroup Classification

The UPEC isolates were assigned into the distinct E. coli phylogroups (A, B1, B2, C, D, E, and F) and clades using a quadruplex PCR [28]. Subsequently, UPEC isolates classified in the phylogroup B2, with the following genotype: arpA−, chuA+, yjaA−, and TspE4+, and all UPEC isolates from the phylogroup F were submitted to a triplex PCR [29] to confirm these isolates as B2 and F, or to reclassify them into the newly described phylogroup G.

2.5. Antimicrobial Resistance Profile and Detection of ESBL-Producing E. coli

Antimicrobial susceptibility assays were performed using the disk diffusion method, performed on Mueller-Hinton agar (OXOID, Basingstoke, UK). The inhibition zone diameter was interpreted following the Clinical and Laboratory Standards Institute [38], except for cephalothin, a first-generation cephalosporin, whose breakpoint was from a previous publication [39].
The 19 antimicrobial drugs tested were: ampicillin (AMP; 10 μg), piperacillin/tazobactam (PPT, 100/10 μg), ampicillin/sulbactam (ASB, 10/10 μg), amoxicillin/clavulanic acid (AMC; 20/10 μg), cephalotin (CFL, 30 μg), cefuroxime (CRX, 30 μg), ceftazidime (CAZ, 30 μg), cefotaxime (CTX, 30 μg), cefepime (CPM, 30 μg), ertapenem (ETP, 10 μg), meropenem (MER, 10 μg), gentamicin (GEN, 10 μg), amikacin (AMI, 30 μg), nalidixic acid (NAL, 30 μg), norfloxacin (NOR, 10 μg), ciprofloxacin (CIP, 5 μg), trimethoprim/sulfamethoxazole (SUT, 1.25/23.75 μg), nitrofurantoin (NIT, 300 μg), and fosfomycin (FOS, 200 μg). All commercial disks impregnated with antimicrobial drugs tested were obtained from Cefar Diagnóstica Ltd. (São Paulo, SP, Brazil), and the E. coli ATCC 25922 was used as quality control from the disks. Multidrug-resistant (MDR) E. coli was defined by the detecting of non-susceptible isolates (resistant or intermediate) to at least one agent of three or more of the six distinct classes of antimicrobial drugs tested, as follows: (1) β-lactams, (2) aminoglycosides, (3) quinolones/fluoroquinolones, (4) folic acid metabolism inhibitors, (5) nitrofuran, and (6) phosphonic acid derivative [40,41].
Extended-spectrum β-lactamase (ESBL) production was investigated using ceftazidime (30 μg/mL) and cefotaxime (30 μg/mL) with and without clavulanic-acid (10 μg/mL). An increase of 5 mm or more in the zone diameter, observed in the presence of clavulanic acid, was considered positive for ESBL production [38].

2.6. Detection of Beta-Lactamase-Encoding Genes

Genes encoding the main ESBL groups, such as TEM (blaTEM-1), SHV (blaSHV), and CTX-M (blaCTX-M-2-group, blaCTX-M-8-group, and blaCTX-M-15-group), as well as the AmpC cephalosporinase CMY-2 (blaCMY-2), were investigated by PCR using primers as described in Table S3.

3. Results

The large majority of outpatients from which the 112 UPEC isolates were obtained was female (90.2%, 101/112), with ages ranging from 18 to 65 years (65.3%, 66/101) (Table S4). Clinically, the outpatients with community acquired UTIs were diagnosed as follows: cystitis (77.7%, 87/112), pyelonephritis (9.8%, 11/112), or asymptomatic bacteriuria (9.8%, 11/112). In addition, three outpatients (2.7%) did not have their diagnosis documented in their medical records (Table S4).
Regarding somatic antigen (O) typing, which defines the E. coli serogroups, 64 UPEC isolates (57.1%, 64/112) were identified in 31 distinct serogroups, with the serogroups O6 (12.5%, 14/112) and O25 (8.9%, 10/112) as the most frequently detected, besides the occurrence of O-non-typeable (28.6%, 32/112) or autoagglutinable (14.3%, 16/112) UPEC isolates (Table S5). Considering the O:H serotypes, O6:H1 and O25:H4 were the most frequent and equally detected in 7.1% (8/112) of the UPEC isolates studied (Table S5).
We observed that a large number of isolates were assigned into the E. coli phylogroup B2 (41.07%, 46/112), followed by isolates assigned in the phylogroups A (16.07%, 18/112), B1 (14.29%, 16/112), D (12.5%, 14/112), F (7.14%, 8/112), and G (3.57%, 4/112). Less commonly, we observed UPEC isolates assigned into the phylogroups C, E and E. clades, with a frequency of 1.79% (2/112) in each of the phylogroups (Table 1).
Of the 29 virulence factor-encoding genes investigated, the most frequently detected were fimH (98.2%, 110/112), traT (81.3%, 91/112), ecpA (77.7%, 87/112), ompT (62.5%, 70/112), irp2 (52.7%, 59/112), and sitA (51.8%, 58/112) (Table 1). Some particularities should be highlighted, such as the occurrence of some genes in just one or two phylogroups, such as sfa/focDE, cnf1, and cdt, exclusively detected in isolates from the phylogroup B2; ibeA only in isolates from the phylogroups B2 and F; and vat only in isolates from the phylogroups B2 and G (Table 1). UPEC with the highest number of virulence factor-encoding genes were observed in isolates assigned to phylogroup B2, with a mean of 9.5, containing UPEC isolates harboring a minimum of three to a maximum of 17 genes (Table 1). A high mean of virulence factor-encoding genes was also observed in the phylogroups D (8.3), G (7.8), and F (6.1) (Table 1).
Five UPEC isolates (4.5%) harbored virulence markers frequently used for diagnosis of two important DEC pathotypes, aEPEC (eae+) and EAEC (aggR+ and aatA+), and thus were designated as hybrid UPEC/aEPEC (one isolate) and UPEC/EAEC (four isolates), respectively (Table 2). The five UPEC/DEC hybrid isolates were obtained from four patients diagnosed with cystitis and one with pyelonephritis; the patients were of both genders (four female and one male), aged from 1 to 84 years (Table 3). Other relevant clinical information from the patients from which the hybrid UPEC/DEC isolates were obtained is summarized in Table 3. All five patients who had UTIs due to hybrid UPEC/DEC isolates progressed to the curing of the infection (data not shown).
These hybrid isolates were heterogeneous in terms of serotype, phylogroup classification and content of genes encoding virulence factors associated with the ExPEC pathogenesis, as shown in Table 2. All five hybrid isolates harbored the fimH, ecpA, traT, and ompT genes, three UPEC/EAEC isolates (75%, 3/4) harbored genes from the pap operon (papA and/or papC) and/or irp2, while the ibeA gene was observed only in the UPEC/aEPEC isolate from the phylogroup B2 (Table 2).
The highest resistance rates were observed for the following antimicrobial drugs: ampicillin (46.4% 52/112) and trimethoprim/sulfamethoxazole (34.8%, 39/112), as well as for the quinolones/fluoroquinolones tested, as follows: nalidixic acid (31.3%, 35/112), ciprofloxacin (31.3%; 35/112) and norfloxacin (20.5%, 23/112) (Table 4). On the other hand, 99.1% (111/112) of the UPEC isolates tested were susceptible to nitrofurantoin and/or fosfomycin (Table 4). Thirty-one isolates (27.7%, 31/112), including two hybrid UPEC/EAEC (isolates 85 and 100), were classified as MDR, with the majority of them (77.4%, 24/31) showing non-susceptibility to the following classes of antibiotics tested concomitantly: β-lactams, quinolones/fluoroquinolones and folic acid metabolism inhibitors (Table S6). Worryingly, 9.82% (11/112) were classified as an ESBL-producing UPEC (Table 5). The majority (72.73%, 8/11) harbored the blaCTX-M-15-group, with this gene detected with the AmpC cephalosporinase-encoding blaCMY-2 gene in two isolates concomitantly (Table 5).
Furthermore, an overview of the resistance profile of the five hybrid isolates identified in this study (four UPEC/EAEC and one UPEC/aEPEC) is summarized in Table 6. We observed that the UPEC/EAEC hybrid isolates were non-susceptible to antimicrobial drugs of the following classes: β-lactams (100.0%, 4/4), aminoglycosides (50.0%, 2/4), quinolones/fluoroquinolones (50.0%, 2/4), and folic acid metabolism inhibitors (50.0%, 2/4). On the other hand, the hybrid UPEC/aEPEC isolate 92 was susceptible to all antimicrobial drugs tested (Table 6). Of note, one hybrid UPEC/EAEC (isolate 85) was identified as an ESBL-producer (Table 6).

4. Discussion

In the present study, we provide an extensive molecular and phenotypic characterization of Brazilian UPEC isolates, such as the classification of UPEC isolates in the distinct E. coli phylogroups, the presence of virulence factor encoding-genes, O:H serotypes, and antimicrobial resistance profiles. Together, we observed that UPEC isolates circulating in the city of Botucatu are very heterogeneous in all features investigated, similar to what was observed in other studies carried out in Brazil, as well as in other countries [31,32,42,43]. Despite the high diversity observed, some characteristics seem to be common when comparing UPEC isolates from distinct geographic regions, such as the majority of the isolates being assigned to phylogroup B2 and serotype O25:H4 as one of the most commonly detected [31,32].
DEC and ExPEC isolates use distinct virulence factors to cause intestinal and extraintestinal diseases in the human host, respectively [2,3]. However, the identification of hybrid E. coli isolates, combining virulence factor encoding-genes from both pathogenic E. coli groups, i.e., DEC and ExPEC, is becoming more and more frequent in the literature [18,20,21,44,45,46,47]. Among the most common DEC molecular markers that have been identified in UPEC, we can highlight the locus of enterocyte effacement (LEE region), a chromosomal PAI from EPEC, and the aggregative adherence plasmid (pAA), from EAEC [18,20,45,47]. Less frequently, UPEC isolates harboring the stx1 and/or stx2 genes, which are molecular markers of Shiga toxin-producing E. coli (STEC) that encode potent cytotoxins that inhibit protein synthesis in eukaryotic cells [48], have also been detected [46,49,50].
The LEE encodes proteins that induce the formation of a histopathological lesion, termed attaching and effacement (AE), in infected epithelial cells [51]. The AE lesion is characterized by intimate adherence of bacteria to the host cell, which is mediated by the interaction between the adhesin intimin and its translocated receptor, microvillus effacement, F-actin accumulation, and the formation of a pedestal-like structure underneath adherent bacteria [52,53]. A recent study has compiled evidence that a hybrid aEPEC/UPEC-252 isolate (serotype O71:H40) harbor all 41 genes from the LEE region, produces the adhesin intimin (encoded by the eae gene located in the LEE region), adheres to both urinary (T24 and HEK 293T) and intestinal (Caco-2 and LS174T) cell lineages, and induces AE lesions in HeLa cells [54]. The latter study may be the first to demonstrate that the LEE region, present in E. coli isolates obtained from extraintestinal sites, can be functional. However, whether the proteins encoded by genes from the LEE region contribute to the establishment, or aggravation, of UTIs has yet to be determined.
The hybrid UPEC/aEPEC-92 (UPEC isolate harboring the LEE region isolated in this study) belongs to serotype O145:H34 and harbored the ibeA gene. This gene was first identified in a meningitis-associated E. coli (MNEC) K1 as part of the GimA (genetic island of meningitis E. coli containing ibeA) PAI, which is organized in four distinct operons (ptnIPKC, cglDTEC, gcxKRCI, and ibeRAT). Of note, IbeA contributes to MNEC K1 crossing the blood-brain barrier [55]. Even though this isolate was first classified as UPEC, due to the site of isolation, and then reclassified as a hybrid UEPC/aEPEC, due to the presence of the LEE region, the presence of the ibeA gene from MNEC cannot be ignored. This type of data brings to light all the complexity in the pathogenic E. coli classification, which is based on the infection site for ExPEC isolates, and in the presence of molecular markers for DEC isolates [56]. In Brazil, aEPEC of serotype O145:H34 has also been isolated from diarrheal patients [12] and, similar to the hybrid UPEC/aEPEC-92, belongs to the phylogroup B2 [57,58]. Together, these data suggest that UPEC and/or aEPEC of serotype O145:H34 may act as a heteropathogen, which can be defined as pathogenic E. coli isolates that occupy an evolutionary interface between ExPEC and DEC (hybrid ExPEC/DEC isolates), thus, being able to cause disease in the intestinal and extraintestinal sites of human hosts [45,49].
Another important genetic element from DEC that has been frequently found in UPEC isolates is the pAA [18,21,44,47], which is characteristic of EAEC isolates. EAEC is defined as E. coli isolates that produce the aggregative adherence pattern (AA) on and around epithelial cells, characterized by a bacterial arrangement that resembles stacked bricks [59]. The pAA harbors an operon that encodes the proteins necessary for the biogenesis of one of the five AAF (aggregative adherence fimbria) types (AAF/I–AAF/V), which is responsible for the establishment of the AA pattern [60,61]; besides, it carries the genes encoding other EAEC-associated virulence factors, such as a type I secretion system (aatPABCD), the anti-aggregation protein dispersin (aap), heat-stable enterotoxin EAST1 (astA), plasmid-encoded toxin (pet), shigella extracellular protein (sepA), as well as a transcriptional activator of virulence genes (aggR) [62]. Studies from the literature have shown that some hybrid UPEC/EAEC isolates can produce the AA pattern on HeLa cells, as well as form biofilm on abiotic surfaces [18,20,45,47], thus demonstrating that some hybrid UPEC/EAEC isolates preserve the ability to produce phenotypes compatible with isolates from the EAEC pathotype.
The pathogenic potential of UPEC/EAEC isolates can be illustrated by an UTI outbreak that occurred in Copenhagen, Denmark, during the year 1991, due to an E. coli strain of serotype O78:H10 that harbored several EAEC virulence-associated genes, such as sat, pic, aatA, aggR, aggA, aaiC, and aap [19]. An additional study demonstrated that C555-91 (a representative isolate from the Copenhagen UTI outbreak) causes urovirulence in a mouse model without the requirement for AAF/I. In addition, biofilm formation in urethral catheters is dependent on this fimbria production [63], thus indicating at which stage of the hybrid UPEC/EAEC infection the EAEC virulence factors are really necessary. There is also a report in the literature of a 55-year-old female patient hospitalized with persistent diarrhea and vomiting who had a UTI followed by sepsis [64]. In this patient, EAEC of serotype O176:HNT was isolated from urine and blood samples, and three types of DEC were isolated from the stool, two of which were EAEC (serotypes O176:HNT and O12:H4) and one aEPEC (O56:H6). Although it seems evident that EAEC of serotype O176:HNT caused the UTI and sepsis, the isolation of three types of DEC from the stool sample makes it challenging to identify the pathogen associated with the diarrheal disease and, consequently, the association of the EAEC O176:HNT as a heteropathogen. By associating the uropathogenic potential of these E. coli isolates (serotypes O78:H10 and O176:HNT) with the presence of EAEC markers, we can extrapolate the information from the studies mentioned above and conclude that these isolates are, in fact, hybrid UPEC/EAEC. Furthermore, EAEC with uropathogenic features are present in the stool of asymptomatic individuals and may be a source of UTI [65].
Isolates of E. coli serotype O3:H2, molecularly classified as EAEC, have been frequently isolated from stool samples from patients with diarrhea, as well as from asymptomatic individuals in studies carried out in Brazil [12,66,67,68,69]. In the present study, the hybrid UPEC/EAEC-34 of serotype O3:H2 was isolated from a UTI, thus suggesting that some bacteria of this serotype may act as a heteropathogen. Except for serotype O3:H2, the other serotypes identified among the UPEC/EAEC isolates in this study (O15:H6, O126:H10, and O90:H2) seem not to be associated with cases of diarrhea caused by EAEC in the Brazilian setting [12,68].
Usually, community-acquired UTI is more responsive to antimicrobial treatment and has lower antimicrobial resistance rates than hospital-acquired UTI [70]. In Brazil, the current consensus recommends the use of fosfomycin and nitrofurantoin as the first choices for the treatment of uncomplicated UTI in women [71], and we found that only one isolate was not susceptible to these drugs in vitro; therefore, empirical recommendations are potentially effective against UPEC in our region.
On the other hand, we detected isolates presenting the MDR phenotype, including ESBL-producing UPEC. We identified CTX-M-8 and CTX-M-15-producing isolates belonging to diversified phylogroups and serotypes, likely indicating multiple acquisitions of these determinants in different evolutionary events. CTX-M-producing E. coli with different genetic backgrounds was already reported in clinical UPEC [34] and in environmental E. coli isolates from the food chain in Brazil [72]. In addition, an increase in the prevalence of ESBL-producing or quinolone-resistant UPEC was reported in Rio de Janeiro [35], indicating that the occurrence of drug- resistant UPEC is disseminated in different regions. Although quinolones and 3rd-generation cephalosporin are not recommended to treat uncomplicated UTI, alternative drugs are needed for other complicated infections, which can represent a potential threat in terms of resistance dissemination and public health implications [35].
Data on the resistance profile of hybrid UPEC/EAEC and UPEC/aEPEC isolates are scarce in the literature. However, a very recently published article characterizing UPEC isolates obtained from 100 ambulatory patients with community-acquired UTIs in the city of Chilpancingo, Mexico, reported that 77.27% (17/22) of the UPEC/EAEC hybrid isolates studied were classified as ESBL producers, besides the high resistance rates observed for β-lactams, aminoglycosides, quinolones/fluoroquinolones, and folic acid metabolism inhibitors [73]. Moreover, a hybrid UPEC/aEPEC identified as an ESBL-producer, and harboring the blaCTX-M-15 gene, was reported as a causative agent of UTI in Brazil [47].

5. Conclusions

Our data reinforce that hybrid UPEC/EAEC and UPEC/aEPEC isolates are circulating in the city of Botucatu, Brazil, as important uropathogens. However, how and whether these specific combinations of genes can influence their pathogenicity is a question that remains to be investigated.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/microorganisms10030645/s1, Table S1: Primers used to detect virulence factor-encoding genes in the uropathogenic E. coli (UPEC) isolates studied [74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91]; Table S2: Primers used to detect virulence factor-encoding genes associated with the distinct diarrheagenic E. coli (DEC) pathotypes [92,93,94,95,96]; Table S3: Primers used to detect β-lactamase-encoding genes in the Extended Spectrum Beta-Lactamases (ESBL)-producing uropathogenic E. coli (UPEC) isolates identified in this study [97,98,99,100,101,102]; Table S4: Age, gender and clinical diagnosis of the outpatients with urinary tract infections included in this study; Table S5: Serotypes and E. coli phylogroups of the uropathogenic E. coli (UPEC) isolates studied; and Table S6: Resistance profile of the 31 uropathogenic E. coli (UPEC) isolates with multidrug-resistance (MDR) phenotype.

Author Contributions

Conceptualization, R.H.S.T., L.F.d.S., T.A.T.G., C.H.C. and R.T.H.; Data curation, R.C.B.D., D.R.P.d.L., A.L.M. and R.T.H.; Formal analysis, R.H.S.T., R.C.B.D., H.O., D.R.P.d.L., M.A.V., L.F.d.S., A.M.F., V.L.M.R., A.L.M. and C.H.C.; Funding acquisition, T.A.T.G. and R.T.H.; Investigation, R.H.S.T., R.C.B.D., H.O., D.R.P.d.L., M.A.V., L.F.d.S., A.M.F., V.L.M.R. and C.H.C.; Methodology, R.H.S.T., R.C.B.D., H.O., D.R.P.d.L., M.A.V., L.F.d.S., A.M.F., V.L.M.R., A.L.M. and C.H.C.; Project administration, L.F.d.S., C.H.C. and R.T.H.; Supervision, R.T.H.; Validation, R.H.S.T., R.C.B.D., L.F.d.S. and A.M.F.; Writing–original draft, C.H.C. and R.T.H.; Writing–review & editing, D.R.P.d.L., A.L.M., T.A.T.G., C.H.C. and R.T.H. All authors have read and agreed to the published version of the manuscript.

Funding

This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-Brasil (CAPES) - Finance Code 001; and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grants 2018/17353-7 to T.A.T.G. and 2015/26207-6 to R.T.H.).

Institutional Review Board Statement

The study was conducted in accordance with the Declaration of Helsinki, and approved by the Botucatu Medical School Ethical Committee for human experimentation (CAAE: 09994419.6.1002.5795).

Informed Consent Statement

All UPEC isolates studied were obtained from clinical routine after laboratory analysis, and clinical information of patients was obtained from anonymized medical records. No additional procedure was performed to acquire any bacterial isolate. Therefore, the study was exempted from any specific written informed consent as determined by the Brazilian National Health Council n° 466/12 and 510/16.

Data Availability Statement

The raw data of this study will be made available by the authors, without reservation, to any qualified researcher.

Acknowledgments

The authors thank Cefar Diagnóstica Ltd. for providing the antimicrobial drug disks used in this study.

Conflicts of Interest

The authors declare that they have no conflict of interest.

References

  1. Nataro, J.P.; Kaper, J.B. Diarrheagenic Escherichia coli. Clin. Microbiol. Rev. 1998, 11, 142–201. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  2. Kaper, J.B.; Nataro, J.P.; Mobley, H.L.T.T. Pathogenic Escherichia coli. Nat. Rev. Microbiol. 2004, 2, 123–140. [Google Scholar] [CrossRef] [PubMed]
  3. Croxen, M.A.; Law, R.J.; Scholz, R.; Keeney, K.M.; Wlodarska, M.; Finlay, B.B. Recent Advances in Understanding Enteric Pathogenic Escherichia coli. Clin. Microbiol. Rev. 2013, 26, 822–880. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  4. Burland, V.; Shao, Y.; Perna, N.T.; Plunkett, G.; Sofia, H.J.; Blattner, F.R. The Complete DNA Sequence and Analysis of the Large Virulence Plasmid of Escherichia coli O157:H7. Nucleic Acids Res. 1998, 26, 4196–4204. [Google Scholar] [CrossRef] [Green Version]
  5. Perna, N.T.; Plunkett, G.; Burland, V.; Mau, B.; Glasner, J.D.; Rose, D.J.; Mayhew, G.F.; Evans, P.S.; Gregor, J.; Kirkpatrick, H.A.; et al. Genome Sequence of Enterohaemorrhagic Escherichia coli O157:H7. Nature 2001, 409, 529–533. [Google Scholar] [CrossRef] [Green Version]
  6. Welch, R.A.; Burland, V.; Plunkett, G.; Redford, P.; Roesch, P.; Rasko, D.; Buckles, E.L.; Liou, S.R.; Boutin, A.; Hackett, J.; et al. Extensive Mosaic Structure Revealed by the Complete Genome Sequence of Uropathogenic Escherichia coli. Proc. Natl. Acad. Sci. USA 2002, 99, 17020–17024. [Google Scholar] [CrossRef] [Green Version]
  7. Iguchi, A.; Thomson, N.R.; Ogura, Y.; Saunders, D.; Ooka, T.; Henderson, I.R.; Harris, D.; Asadulghani, M.; Kurokawa, K.; Dean, P.; et al. Complete Genome Sequence and Comparative Genome Analysis of Enteropathogenic Escherichia coli O127:H6 Strain E2348/69. J. Bacteriol. 2009, 191, 347–354. [Google Scholar] [CrossRef] [Green Version]
  8. Chaudhuri, R.R.; Sebaihia, M.; Hobman, J.L.; Webber, M.A.; Leyton, D.L.; Goldberg, M.D.; Cunningham, A.F.; Scott-Tucker, A.; Ferguson, P.R.; Thomas, C.M.; et al. Complete Genome Sequence and Comparative Metabolic Profiling of the Prototypical Enteroaggregative Escherichia coli Strain 042. PLoS ONE 2010, 5, e8801. [Google Scholar] [CrossRef]
  9. Cointe, A.; Birgy, A.; Mariani-Kurkdjian, P.; Liguori, S.; Courroux, C.; Blanco, J.; Delannoy, S.; Fach, P.; Loukiadis, E.; Bidet, P.; et al. Emerging Multidrug-Resistant Hybrid Pathotype Shiga Toxin–Producing Escherichia coli O80 and Related Strains of Clonal Complex 165, Europe. Emerg. Infect. Dis. 2018, 24, 2262–2269. [Google Scholar] [CrossRef] [Green Version]
  10. Russo, T.A.; Johnson, J.R. Proposal for a New Inclusive Designation for Extraintestinal Pathogenic Isolates of Escherichia coli: ExPEC. J. Infect. Dis. 2000, 181, 1753–1754. [Google Scholar] [CrossRef] [Green Version]
  11. Hernandes, R.T.; Elias, W.P.; Vieira, M.A.M.; Gomes, T.A.T. An Overview of Atypical Enteropathogenic Escherichia coli. FEMS Microbiol. Lett. 2009, 297, 137–149. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  12. Ori, E.L.; Takagi, E.H.; Andrade, T.S.; Miguel, B.T.; Cergole-Novella, M.C.; Guth, B.E.C.; Hernandes, R.T.; Dias, R.C.B.; Pinheiro, S.R.S.; Camargo, C.H.; et al. Diarrhoeagenic Escherichia coli and Escherichia albertii in Brazil: Pathotypes and Serotypes over a 6-Year Period of Surveillance. Epidemiol. Infect. 2019, 147, 1–9. [Google Scholar] [CrossRef] [Green Version]
  13. Lima, A.A.M.; Oliveira, D.B.; Quetz, J.S.; Havt, A.; Prata, M.M.G.; Lima, I.F.N.; Soares, A.M.; Filho, J.Q.; Lima, N.L.; Medeiros, P.H.Q.S.; et al. Etiology and Severity of Diarrheal Diseases in Infants at the Semiarid Region of Brazil: A Case-Control Study. PLoS Negl. Trop. Dis. 2019, 13, e0007154. [Google Scholar] [CrossRef] [PubMed]
  14. Kotloff, K.L.; Nataro, J.P.; Blackwelder, W.C.; Nasrin, D.; Farag, T.H.; Panchalingam, S.; Wu, Y.; Sow, S.O.; Sur, D.; Breiman, R.F.; et al. Burden and Aetiology of Diarrhoeal Disease in Infants and Young Children in Developing Countries (the Global Enteric Multicenter Study, GEMS): A Prospective, Case-Control Study. Lancet 2013, 382, 209–222. [Google Scholar] [CrossRef]
  15. Araujo, J.M.; Tabarelli, G.F.; Aranda, K.R.S.; Fabbricotti, S.H.; Fagundes-Neto, U.; Mendes, C.M.F.; Scaletsky, I.C.A. Typical Enteroaggregative and Atypical Enteropathogenic Types of Escherichia coli Are the Most Prevalent Diarrhea-Associated Pathotypes among Brazilian Children. J. Clin. Microbiol. 2007, 45, 3396–3399. [Google Scholar] [CrossRef] [Green Version]
  16. Bueris, V.; Sircili, M.P.; Taddei, C.R.; dos Santos, M.F.; Franzolin, M.R.; Martinez, M.B.; Ferrer, S.R.; Barreto, M.L.; Trabulsi, L.R. Detection of Diarrheagenic Escherichia coli from Children with and without Diarrhea in Salvador, Bahia, Brazil. Mem. Inst. Oswaldo Cruz 2007, 102, 839–844. [Google Scholar] [CrossRef]
  17. Dias, R.C.B.; Dos Santos, B.C.; Dos Santos, L.F.; Vieira, M.A.; Yamatogi, R.S.; Mondelli, A.L.; Sadatsune, T.; Sforcin, J.M.; Gomes, T.A.T.; Hernandes, R.T. Diarrheagenic Escherichia coli Pathotypes Investigation Revealed Atypical Enteropathogenic E. coli as Putative Emerging Diarrheal Agents in Children Living in Botucatu, São Paulo State, Brazil. APMIS 2016, 124, 299–308. [Google Scholar] [CrossRef]
  18. Abe, C.M.; Salvador, F.A.; Falsetti, I.N.; Vieira, M.A.M.; Blanco, J.; Blanco, J.E.; Blanco, M.; Machado, A.M.O.; Elias, W.P.; Hernandes, R.T.; et al. Uropathogenic Escherichia coli (UPEC) Strains May Carry Virulence Properties of Diarrhoeagenic E. coli. FEMS Immunol. Med. Microbiol. 2008, 52, 397–406. [Google Scholar] [CrossRef] [Green Version]
  19. Olesen, B.; Scheutz, F.; Andersen, R.L.; Menard, M.; Boisen, N.; Johnston, B.; Hansen, D.S.; Krogfelt, K.A.; Nataro, J.P.; Johnson, J.R. Enteroaggregative Escherichia coli O78:H10, the Cause of an Outbreak of Urinary Tract Infection. J. Clin. Microbiol. 2012, 50, 3703–3711. [Google Scholar] [CrossRef] [Green Version]
  20. Lara, F.B.M.; Nery, D.R.; de Oliveira, P.M.; Araujo, M.L.; Carvalho, F.R.Q.; Messias-Silva, L.C.F.; Ferreira, L.B.; Faria-Junior, C.; Pereira, A.L. Virulence Markers and Phylogenetic Analysis of Escherichia coli Strains with Hybrid EAEC/UPEC Genotypes Recovered from Sporadic Cases of Extraintestinal Infections. Front. Microbiol. 2017, 8, 146. [Google Scholar] [CrossRef] [Green Version]
  21. Boll, E.J.; Overballe-Petersen, S.; Hasman, H.; Roer, L.; Ng, K.; Scheutz, F.; Hammerum, A.M.; Dungu, A.; Hansen, F.; Johannesen, T.B.; et al. Emergence of Enteroaggregative Escherichia coli within the ST131 Lineage as a Cause of Extraintestinal Infections. MBio 2020, 11, e00353-20. [Google Scholar] [CrossRef] [PubMed]
  22. Mandomando, I.; Vubil, D.; Boisen, N.; Quintó, L.; Ruiz, J.; Sigaúque, B.; Nhampossa, T.; Garrine, M.; Massora, S.; Aide, P.; et al. Escherichia coli ST131 Clones Harbouring AggR and AAF/V Fimbriae Causing Bacteremia in Mozambican Children: Emergence of New Variant of fimH27 Subclone. PLoS Negl. Trop. Dis. 2020, 14, e0008274. [Google Scholar] [CrossRef] [PubMed]
  23. Foxman, B. The Epidemiology of Urinary Tract Infection. Nat. Rev. Urol. 2010, 7, 653–660. [Google Scholar] [CrossRef]
  24. Nielubowicz, G.R.; Mobley, H.L.T. Host–Pathogen Interactions in Urinary Tract Infection. Nat. Rev. Urol. 2010, 7, 430–441. [Google Scholar] [CrossRef] [PubMed]
  25. Subashchandrabose, S.; Mobley, H.L.T. Virulence and Fitness Determinants of Uropathogenic Escherichia coli. Microbiol. Spectr. 2015, 3, 235–261. [Google Scholar] [CrossRef] [Green Version]
  26. Dobrindt, U.; Blum-Oehler, G.; Nagy, G.; Schneider, G.; Johann, A.; Gottschalk, G.; Hacker, J. Genetic Structure and Distribution of Four Pathogenicity Islands (PAI I536 to PAI IV536) of Uropathogenic Escherichia coli Strain 536. Infect. Immun. 2002, 70, 6365–6372. [Google Scholar] [CrossRef] [Green Version]
  27. Lloyd, A.L.; Rasko, D.A.; Mobley, H.L.T. Defining Genomic Islands and Uropathogen-Specific Genes in Uropathogenic Escherichia coli. J. Bacteriol. 2007, 189, 3532–3546. [Google Scholar] [CrossRef] [Green Version]
  28. Clermont, O.; Christenson, J.K.; Denamur, E.; Gordon, D.M. The Clermont Escherichia coli Phylo-Typing Method Revisited: Improvement of Specificity and Detection of New Phylo-Groups. Environ. Microbiol. Rep. 2013, 5, 58–65. [Google Scholar] [CrossRef]
  29. Clermont, O.; Dixit, O.V.A.; Vangchhia, B.; Condamine, B.; Dion, S.; Bridier-Nahmias, A.; Denamur, E.; Gordon, D. Characterization and Rapid Identification of Phylogroup G in Escherichia coli, a Lineage with High Virulence and Antibiotic Resistance Potential. Environ. Microbiol. 2019, 21, 3107–3117. [Google Scholar] [CrossRef]
  30. Denamur, E.; Clermont, O.; Bonacorsi, S.; Gordon, D. The Population Genetics of Pathogenic Escherichia coli. Nat. Rev. Microbiol. 2021, 19, 37–54. [Google Scholar] [CrossRef]
  31. Campos, A.C.C.; Andrade, N.L.; Ferdous, M.; Chlebowicz, M.A.; Santos, C.C.; Correal, J.C.D.; Lo Ten Foe, J.R.; Rosa, A.C.P.; Damasco, P.V.; Friedrich, A.W.; et al. Comprehensive Molecular Characterization of Escherichia coli Isolates from Urine Samples of Hospitalized Patients in Rio de Janeiro, Brazil. Front. Microbiol. 2018, 9, 243. [Google Scholar] [CrossRef] [PubMed]
  32. Flament-Simon, S.-C.; Nicolas-Chanoine, M.-H.; García, V.; Duprilot, M.; Mayer, N.; Alonso, M.P.; García-Meniño, I.; Blanco, J.E.; Blanco, M.; Blanco, J. Clonal Structure, Virulence Factor-Encoding Genes and Antibiotic Resistance of Escherichia coli, Causing Urinary Tract Infections and Other Extraintestinal Infections in Humans in Spain and France during 2016. Antibiotics 2020, 9, 161. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  33. Tamadonfar, K.O.; Omattage, N.S.; Spaulding, C.N.; Hultgren, S.J. Reaching the End of the Line: Urinary Tract Infections. Microbiol. Spectr. 2019, 7, 83–99. [Google Scholar] [CrossRef] [PubMed]
  34. Gonçalves, L.F.; de Oliveira Martins-Júnior, P.; de Melo, A.B.F.; da Silva, R.C.R.M.; de Paulo Martins, V.; Pitondo-Silva, A.; de Campos, T.A. Multidrug Resistance Dissemination by Extended-Spectrum β-Lactamase-Producing Escherichia coli Causing Community-Acquired Urinary Tract Infection in the Central-Western Region, Brazil. J. Glob. Antimicrob. Resist. 2016, 6, 1–4. [Google Scholar] [CrossRef] [PubMed]
  35. Da-Silva, A.P.d.S.; de Sousa, V.S.; Longo, L.G.d.A.; Caldera, S.; Baltazar, I.C.L.; Bonelli, R.R.; Santoro-Lopes, G.; Riley, L.W.; Moreira, B.M. Prevalence of Fluoroquinolone-Resistant and Broad-Spectrum Cephalosporin-Resistant Community-Acquired Urinary Tract Infections in Rio de Janeiro: Impact of Escherichia coli Genotypes ST69 and ST131. Infect. Genet. Evol. 2020, 85, 104452. [Google Scholar] [CrossRef]
  36. Dias, R.C.B.; Vieira, M.A.; Moro, A.C.; Ribolli, D.F.M.; Monteiro, A.C.M.; Camargo, C.H.; Tiba-Casas, M.R.; Soares, F.B.; dos Santos, L.F.; Montelli, A.C.; et al. Characterization of Escherichia coli Obtained from Patients Undergoing Peritoneal Dialysis and Diagnosed with Peritonitis in a Brazilian Centre. J. Med. Microbiol. 2019, 68, 1330–1340. [Google Scholar] [CrossRef]
  37. Ewing, W.H. Edwards and Ewing’s Identification of Enterobacteriaceae, 4th ed.; Elsevier Science Publishing: New York, NY, USA, 1986. [Google Scholar]
  38. CLSI. Performance Standards for Antimicrobial Susceptibility Testing, 30th ed.; CLSI Supplement M100; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2020. [Google Scholar]
  39. CLSI. Performance Standards for Antimicrobial Susceptibility Testing, 25th ed.; CLSI Supplement M100-S25; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2015. [Google Scholar]
  40. Magiorakos, A.-P.; Srinivasan, A.; Carey, R.B.; Carmeli, Y.; Falagas, M.E.; Giske, C.G.; Harbarth, S.; Hindler, J.F.; Kahlmeter, G.; Olsson-Liljequist, B.; et al. Multidrug-Resistant, Extensively Drug-Resistant and Pandrug-Resistant Bacteria: An International Expert Proposal for Interim Standard Definitions for Acquired Resistance. Clin. Microbiol. Infect. 2012, 18, 268–281. [Google Scholar] [CrossRef] [Green Version]
  41. Chattaway, M.A.; Day, M.; Mtwale, J.; White, E.; Rogers, J.; Day, M.; Powell, D.; Ahmad, M.; Harris, R.; Talukder, K.A.; et al. Clonality, Virulence and Antimicrobial Resistance of Enteroaggregative Escherichia coli from Mirzapur, Bangladesh. J. Med. Microbiol. 2017, 66, 1429–1435. [Google Scholar] [CrossRef] [Green Version]
  42. Nüesch-Inderbinen, M.T.; Baschera, M.; Zurfluh, K.; Hächler, H.; Nüesch, H.; Stephan, R. Clonal Diversity, Virulence Potential and Antimicrobial Resistance of Escherichia coli Causing Community Acquired Urinary Tract Infection in Switzerland. Front. Microbiol. 2017, 8, 2334. [Google Scholar] [CrossRef] [Green Version]
  43. Dadi, B.R.; Abebe, T.; Zhang, L.; Mihret, A.; Abebe, W.; Amogne, W. Distribution of Virulence Genes and Phylogenetics of Uropathogenic Escherichia coli among Urinary Tract Infection Patients in Addis Ababa, Ethiopia. BMC Infect. Dis. 2020, 20, 108. [Google Scholar] [CrossRef] [Green Version]
  44. Nazemi, A.; Mirinargasi, M.; Merikhi, N.; Sharifi, S.H. Distribution of Pathogenic Genes aatA, aap, aggR, among Uropathogenic Escherichia coli (UPEC) and Their Linkage with StbA Gene. Indian J. Microbiol. 2011, 51, 355–358. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  45. Toval, F.; Köhler, C.-D.; Vogel, U.; Wagenlehner, F.; Mellmann, A.; Fruth, A.; Schmidt, M.A.; Karch, H.; Bielaszewska, M.; Dobrindt, U. Characterization of Escherichia coli Isolates from Hospital Inpatients or Outpatients with Urinary Tract Infection. J. Clin. Microbiol. 2014, 52, 407–418. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  46. Gati, N.S.; Middendorf-Bauchart, B.; Bletz, S.; Dobrindt, U.; Mellmann, A. Origin and Evolution of Hybrid Shiga Toxin-Producing and Uropathogenic Escherichia coli Strains of Sequence Type 141. J. Clin. Microbiol. 2019, 58, 1–16. [Google Scholar] [CrossRef] [PubMed]
  47. Nascimento, J.A.S.; Santos, F.F.; Valiatti, T.B.; Santos-Neto, J.F.; Santos, A.C.M.; Cayô, R.; Gales, A.C.; Gomes, T.A.T. Frequency and Diversity of Hybrid Escherichia coli Strains Isolated from Urinary Tract Infections. Microorganisms 2021, 9, 693. [Google Scholar] [CrossRef]
  48. Karch, H.; Tarr, P.I.; Bielaszewska, M. Enterohaemorrhagic Escherichia coli in Human Medicine. Int. J. Med. Microbiol. 2005, 295, 405–418. [Google Scholar] [CrossRef]
  49. Bielaszewska, M.; Schiller, R.; Lammers, L.; Bauwens, A.; Fruth, A.; Middendorf, B.; Schmidt, M.A.; Tarr, P.I.; Dobrindt, U.; Karch, H.; et al. Heteropathogenic Virulence and Phylogeny Reveal Phased Pathogenic Metamorphosis in Escherichia coli O2:H6. EMBO Mol. Med. 2014, 6, 347–357. [Google Scholar] [CrossRef]
  50. Toval, F.; Schiller, R.; Meisen, I.; Putze, J.; Kouzel, I.U.; Zhang, W.; Karch, H.; Bielaszewska, M.; Mormann, M.; Müthing, J.; et al. Characterization of Urinary Tract Infection-Associated Shiga Toxin-Producing Escherichia coli. Infect. Immun. 2014, 82, 4631–4642. [Google Scholar] [CrossRef] [Green Version]
  51. McDaniel, T.K.; Jarvis, K.G.; Donnenberg, M.S.; Kaper, J.B. A Genetic Locus of Enterocyte Effacement Conserved among Diverse Enterobacterial Pathogens. Proc. Natl. Acad. Sci. USA 1995, 92, 1664–1668. [Google Scholar] [CrossRef] [Green Version]
  52. Moon, H.W.; Whipp, S.C.; Argenzio, R.A.; Levine, M.M.; Giannella, R.A. Attaching and Effacing Activities of Rabbit and Human Enteropathogenic Escherichia coli in Pig and Rabbit Intestines. Infect. Immun. 1983, 41, 1340–1351. [Google Scholar] [CrossRef] [Green Version]
  53. Knutton, S.; Baldwin, T.; Williams, P.H.; McNeish, A.S. Actin Accumulation at Sites of Bacterial Adhesion to Tissue Culture Cells: Basis of a New Diagnostic Test for Enteropathogenic and Enterohemorrhagic Escherichia coli. Infect. Immun. 1989, 57, 1290–1298. [Google Scholar] [CrossRef] [Green Version]
  54. Valiatti, T.B.; Santos, F.F.; Santos, A.C.M.; Nascimento, J.A.S.; Silva, R.M.; Carvalho, E.; Sinigaglia, R.; Gomes, T.A.T. Genetic and Virulence Characteristics of a Hybrid Atypical Enteropathogenic and Uropathogenic Escherichia coli (aEPEC/UPEC) Strain. Front. Cell. Infect. Microbiol. 2020, 10, 492. [Google Scholar] [CrossRef] [PubMed]
  55. Huang, S.-H.; Chen, Y.-H.; Kong, G.; Chen, S.H.M.; Besemer, J.; Borodovsky, M.; Jong, A. A Novel Genetic Island of Meningitic Escherichia coli K1 Containing the ibeA Invasion Gene (GimA): Functional Annotation and Carbon-Source-Regulated Invasion of Human Brain Microvascular Endothelial Cells. Funct. Integr. Genom. 2001, 1, 312–322. [Google Scholar] [CrossRef] [PubMed]
  56. Santos, A.C.M.; Santos, F.F.; Silva, R.M.; Gomes, T.A.T. Diversity of Hybrid- and Hetero-Pathogenic Escherichia coli and Their Potential Implication in More Severe Diseases. Front. Cell. Infect. Microbiol. 2020, 10, 339. [Google Scholar] [CrossRef] [PubMed]
  57. Vieira, M.A.; dos Santos, L.F.; Dias, R.C.B.; Camargo, C.H.; Pinheiro, S.R.S.; Gomes, T.A.T.; Hernandes, R.T. Atypical Enteropathogenic Escherichia coli as Aetiologic Agents of Sporadic and Outbreak-Associated Diarrhoea in Brazil. J. Med. Microbiol. 2016, 65, 998–1006. [Google Scholar] [CrossRef] [PubMed]
  58. Hernandes, R.T.; Hazen, T.H.; dos Santos, L.F.; Richter, T.K.S.; Michalski, J.M.; Rasko, D.A. Comparative Genomic Analysis Provides Insight into the Phylogeny and Virulence of Atypical Enteropathogenic Escherichia coli Strains from Brazil. PLoS Negl. Trop. Dis. 2020, 14, e0008373. [Google Scholar] [CrossRef]
  59. Nataro, J.P.; Kaper, J.B.; Robins-Browne, R.; Prado, V.; Vial, P.; Levine, M.M. Patterns of Adherence of Diarrheagenic Escherichia coli to HEp-2 Cells. Pediatr. Infect. Dis. J. 1987, 6, 829–831. [Google Scholar] [CrossRef]
  60. Jønsson, R.; Liu, B.; Struve, C.; Yang, Y.; Jørgensen, R.; Xu, Y.; Jenssen, H.; Krogfelt, K.A.; Matthews, S. Structural and Functional Studies of Escherichia coli Aggregative Adherence Fimbriae (AAF/V) Reveal a Deficiency in Extracellular Matrix Binding. Biochim. Biophys. Acta. Proteins Proteom. 2017, 1865, 304–311. [Google Scholar] [CrossRef]
  61. Boisen, N.; Østerlund, M.T.; Joensen, K.G.; Santiago, A.E.; Mandomando, I.; Cravioto, A.; Chattaway, M.A.; Gonyar, L.A.; Overballe-Petersen, S.; Stine, O.C.; et al. Redefining Enteroaggregative Escherichia coli (EAEC): Genomic Characterization of Epidemiological EAEC Strains. PLoS Negl. Trop. Dis. 2020, 14, e0008613. [Google Scholar] [CrossRef]
  62. Hebbelstrup Jensen, B.; Olsen, K.E.P.P.; Struve, C.; Krogfelt, K.A.; Petersen, A.M. Epidemiology and Clinical Manifestations of Enteroaggregative Escherichia coli. Clin. Microbiol. Rev. 2014, 27, 614–630. [Google Scholar] [CrossRef] [Green Version]
  63. Boll, E.J.; Struve, C.; Boisen, N.; Olesen, B.; Stahlhut, S.G.; Krogfelt, K.A. Role of Enteroaggregative Escherichia coli Virulence Factors in Uropathogenesis. Infect. Immun. 2013, 81, 1164–1171. [Google Scholar] [CrossRef] [Green Version]
  64. Herzog, K.; Engeler Dusel, J.; Hugentobler, M.; Beutin, L.; Sägesser, G.; Stephan, R.; Hächler, H.; Nüesch-Inderbinen, M. Diarrheagenic Enteroaggregative Escherichia coli Causing Urinary Tract Infection and Bacteremia Leading to Sepsis. Infection 2014, 42, 441–444. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  65. Nunes, K.O.; Santos, A.C.P.; Bando, S.Y.; Silva, R.M.; Gomes, T.A.T.; Elias, W.P. Enteroaggregative Escherichia coli with Uropathogenic Characteristics Are Present in Feces of Diarrheic and Healthy Children. Pathog. Dis. 2017, 75, 1–4. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  66. Zamboni, A.; Fabbricotti, S.H.; Fagundes-Neto, U.; Scaletsky, I.C.A. Enteroaggregative Escherichia coli Virulence Factors Are Found To Be Associated with Infantile Diarrhea in Brazil. J. Clin. Microbiol. 2004, 42, 1058–1063. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  67. França, F.L.S.S.; Wells, T.J.; Browning, D.F.; Nogueira, R.T.; Sarges, F.S.; Pereira, A.C.; Cunningham, A.F.; Lucheze, K.; Rosa, A.C.P.; Henderson, I.R.; et al. Genotypic and Phenotypic Characterisation of Enteroaggregative Escherichia coli from Children in Rio de Janeiro, Brazil. PLoS ONE 2013, 8, e69971. [Google Scholar] [CrossRef] [PubMed]
  68. Dias, R.C.B.; Tanabe, R.H.S.; Vieira, M.A.; Cergole-Novella, M.C.; dos Santos, L.F.; Gomes, T.A.T.; Elias, W.P.; Hernandes, R.T. Analysis of the Virulence Profile and Phenotypic Features of Typical and Atypical Enteroaggregative Escherichia coli (EAEC) Isolated From Diarrheal Patients in Brazil. Front. Cell. Infect. Microbiol. 2020, 10, 144. [Google Scholar] [CrossRef] [Green Version]
  69. de Lira, D.R.P.; Cavalcanti, A.M.F.; Pinheiro, S.R.S.; Orsi, H.; dos Santos, L.F.; Hernandes, R.T. Identification of a Hybrid Atypical Enteropathogenic and Enteroaggregative Escherichia coli (aEPEC/EAEC) Clone of Serotype O3:H2 Associated with a Diarrheal Outbreak in Brazil. Braz. J. Microbiol. 2021, 52, 2075–2079. [Google Scholar] [CrossRef]
  70. Horcajada, J.P.; Shaw, E.; Padilla, B.; Pintado, V.; Calbo, E.; Benito, N.; Gamallo, R.; Gozalo, M.; Rodríguez-Baño, J. Healthcare-Associated, Community-Acquired and Hospital-Acquired Bacteraemic Urinary Tract Infections in Hospitalized Patients: A Prospective Multicentre Cohort Study in the Era of Antimicrobial Resistance. Clin. Microbiol. Infect. 2013, 19, 962–968. [Google Scholar] [CrossRef] [Green Version]
  71. de Rossi, P.; Cimerman, S.; Truzzi, J.C.; Cunha, C.A.d.; Mattar, R.; Martino, M.D.V.; Hachul, M.; Andriolo, A.; Vasconcelos Neto, J.A.; Pereira-Correia, J.A.; et al. Joint Report of SBI (Brazilian Society of Infectious Diseases), FEBRASGO (Brazilian Federation of Gynecology and Obstetrics Associations), SBU (Brazilian Society of Urology) and SBPC/ML (Brazilian Society of Clinical Pathology/Laboratory Medicine): Recommendations for the clinical management of lower urinary tract infections in pregnant and non-pregnant women. Braz. J. Infect. Dis. 2020, 24, 110–119. [Google Scholar] [CrossRef]
  72. Botelho, L.A.B.; Kraychete, G.B.; Costa e Silva, J.L.; Regis, D.V.V.; Picão, R.C.; Moreira, B.M.; Bonelli, R.R. Widespread Distribution of CTX-M and Plasmid-Mediated AmpC β-Lactamases in Escherichia coli from Brazilian Chicken Meat. Mem. Inst. Oswaldo Cruz 2015, 110, 249–254. [Google Scholar] [CrossRef]
  73. Martínez-Santos, V.I.; Ruíz-Rosas, M.; Ramirez-Peralta, A.; Zaragoza García, O.; Resendiz-Reyes, L.A.; Romero-Pineda, O.J.; Castro-Alarcón, N. Enteroaggregative Escherichia coli Is Associated with Antibiotic Resistance and Urinary Tract Infection Symptomatology. PeerJ 2021, 9, e11726. [Google Scholar] [CrossRef]
  74. Johnson, J.R.; Stell, A.L. Extended Virulence Genotypes of Escherichia coli Strains from Patients with Urosepsis in Relation to Phylogeny and Host Compromise. J. Infect. Dis. 2000, 181, 261–272. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  75. Le Bouguenec, C.; Archambaud, M.; Labigne, A. Rapid and Specific Detection of the pap, afa, and sfa Adhesin-Encoding Operons in Uropathogenic Escherichia coli Strains by Polymerase Chain Reaction. J. Clin. Microbiol. 1992, 30, 1189–1193. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  76. Hernandes, R.T.; Velsko, I.; Sampaio, S.C.F.; Elias, W.P.; Robins-Browne, R.M.; Gomes, T.A.T.; Girón, J.A. Fimbrial Adhesins Produced by Atypical Enteropathogenic Escherichia coli Strains. Appl. Environ. Microbiol. 2011, 77, 8391–8399. [Google Scholar] [CrossRef] [Green Version]
  77. Szalo, I.M.; Goffaux, F.; Pirson, V.; Piérard, D.; Ball, H.; Mainil, J. Presence in Bovine Enteropathogenic (EPEC) and Enterohaemorrhagic (EHEC) Escherichia coli of Genes Encoding for Putative Adhesins of Human EHEC Strains. Res. Microbiol. 2002, 153, 653–658. [Google Scholar] [CrossRef]
  78. Ewers, C.; Li, G.; Wilking, H.; Kießling, S.; Alt, K.; Antáo, E.M.; Laturnus, C.; Diehl, I.; Glodde, S.; Homeier, T.; et al. Avian Pathogenic, Uropathogenic, and Newborn Meningitis-Causing Escherichia coli: How Closely Related Are They? Int. J. Med. Microbiol. 2007, 297, 163–176. [Google Scholar] [CrossRef] [PubMed]
  79. Yamamoto, S.; Terai, A.; Yuri, K.; Kurazono, H.; Takeda, Y.; Yoshida, O. Detection of Urovirulence Factors in Escherichia coli by Multiplex Polymerase Chain Reaction. FEMS Immunol. Med. Microbiol. 1995, 12, 85–90. [Google Scholar] [CrossRef]
  80. Tennant, S.M.; Tauschek, M.; Azzopardi, K.; Bigham, A.; Bennett-Wood, V.; Hartland, E.L.; Qi, W.; Whittam, T.S.; Robins-Browne, R.M. Characterisation of Atypical Enteropathogenic E. coli Strains of Clinical Origin. BMC Microbiol. 2009, 9, 117. [Google Scholar] [CrossRef] [Green Version]
  81. Boisen, N.; Ruiz-Perez, F.; Scheutz, F.; Krogfelt, K.A.; Nataro, J.P. High Prevalence of Serine Protease Autotransporter Cytotoxins among Strains of Enteroaggregative Escherichia coli. Am. J. Trop. Med. Hyg. 2009, 80, 294–301. [Google Scholar] [CrossRef] [Green Version]
  82. Momtaz, H.; Karimian, A.; Madani, M.; Safarpoor Dehkordi, F.; Ranjbar, R.; Sarshar, M.; Souod, N. Uropathogenic Escherichia coli in Iran: Serogroup Distributions, Virulence Factors and Antimicrobial Resistance Properties. Ann. Clin. Microbiol. Antimicrob. 2013, 12, 8. [Google Scholar] [CrossRef] [Green Version]
  83. Parham, N.J.; Srinivasan, U.; Desvaux, M.; Foxman, B.; Marrs, C.F.; Henderson, I.R. PicU, a Second Serine Protease Autotransporter of Uropathogenic Escherichia coli. FEMS Microbiol. Lett. 2004, 230, 73–83. [Google Scholar] [CrossRef]
  84. Ewers, C.; Janßen, T.; Kießling, S.; Philipp, H.C.; Wieler, L.H. Molecular Epidemiology of Avian Pathogenic Escherichia coli (APEC) Isolated from Colisepticemia in Poultry. Vet. Microbiol. 2004, 104, 91–101. [Google Scholar] [CrossRef] [PubMed]
  85. Johnson, T.J.; Siek, K.E.; Johnson, S.J.; Nolan, L.K. DNA Sequence of a ColV Plasmid and Prevalence of Selected Plasmid-Encoded Virulence Genes among Avian Escherichia coli Strains. J. Bacteriol. 2006, 188, 745–758. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  86. Johnson, J.R.; Russo, T.A.; Tarr, P.I.; Carlino, U.; Bilge, S.S.; Vary, J.C.; Stell, A.L. Molecular Epidemiological and Phylogenetic Associations of Two Novel Putative Virulence Genes, iha and iroNE. coli, among Escherichia coli Isolates from Patients with Urosepsis. Infect. Immun. 2000, 68, 3040–3047. [Google Scholar] [CrossRef] [Green Version]
  87. Czeczulin, J.R.; Whittam, T.S.; Henderson, I.R.; Navarro-Garcia, F.; Nataro, J.P. Phylogenetic Analysis of Enteroaggregative and Diffusely Adherent Escherichia coli. Infect. Immun. 1999, 67, 2692–2699. [Google Scholar] [CrossRef] [Green Version]
  88. Dezfulian, H.; Batisson, I.; Fairbrother, J.M.; Lau, P.C.K.; Nassar, A.; Szatmari, G.; Harel, J. Presence and Characterization of Extraintestinal Pathogenic Escherichia coli Virulence Genes in F165-Positive E. coli Strains Isolated from Diseased Calves and Pigs. J. Clin. Microbiol. 2003, 41, 1375–1385. [Google Scholar] [CrossRef] [Green Version]
  89. Rodriguez-Siek, K.E.; Giddings, C.W.; Doetkott, C.; Johnson, T.J.; Nolan, L.K. Characterizing the APEC Pathotype. Vet. Res. 2005, 36, 241–256. [Google Scholar] [CrossRef] [Green Version]
  90. Johnson, J.R.; O’Bryan, T.T.; Low, D.A.; Ling, G.; Delavari, P.; Fasching, C.; Russo, T.A.; Carlino, U.; Stell, A.L. Evidence of Commonality between Canine and Human Extraintestinal Pathogenic Escherichia coli Strains That Express PapG Allele III. Infect. Immun. 2000, 68, 3327–3336. [Google Scholar] [CrossRef] [Green Version]
  91. Ewers, C.; Janßen, T.; Kießling, S.; Philipp, H.C.; Wieler, L.H. Rapid Detection of Virulence-Associated Genes in Avian Pathogenic Escherichia coli by Multiplex Polymerase Chain Reaction. Avian Dis. 2005, 49, 269–273. [Google Scholar] [CrossRef]
  92. Hernandes, R.T.; De la Cruz, M.A.; Yamamoto, D.; Girón, J.A.; Gomes, T.A.T. Dissection of the Role of Pili and Type 2 and 3 Secretion Systems in Adherence and Biofilm Formation of an Atypical Enteropathogenic Escherichia coli Strain. Infect. Immun. 2013, 81, 3793–3802. [Google Scholar] [CrossRef] [Green Version]
  93. Müller, D.; Greune, L.; Heusipp, G.; Karch, H.; Fruth, A.; Tschäpe, H.; Schmidt, M.A. Identification of Unconventional Intestinal Pathogenic Escherichia coli Isolates Expressing Intermediate Virulence Factor Profiles by Using a Novel Single-Step Multiplex PCR. Appl. Environ. Microbiol. 2007, 73, 3380–3390. [Google Scholar] [CrossRef] [Green Version]
  94. Yamasaki, S.; Lin, Z.; Shirai, H.; Terai, A.; Oku, Y.; Ito, H.; Ohmura, M.; Karasawa, T.; Tsukamoto, T.; Kurazono, H.; et al. Typing of Verotoxins by DNA Colony Hybridization with Poly- and Oligonucleotide Probes, a Bead-Enzyme-Linked Immunosorbent Assay, and Polymerase Chain Reaction. Microbiol. Immunol. 1996, 40, 345–352. [Google Scholar] [CrossRef] [Green Version]
  95. Schmidt, H.; Knop, C.; Franke, S.; Aleksic, S.; Heesemann, J.; Schmidt, H.; Knop, C.; Franke, S.; Aleksic, S.; Heesemann, R.; et al. Development of PCR for Screening of Enteroaggregative Escherichia coli. J. Clin. Microbiol. 1995, 33, 701–705. [Google Scholar] [CrossRef] [Green Version]
  96. Ratchtrachenchai, O.-A.; Subpasu, S.; Ito, K. Investigation on Enteroaggregative Escherichia coli Infection by Multiplex PCR. Bull. Dep. Med. Sci. 1997, 39, 211–220. [Google Scholar]
  97. Yagi, T.; Kurokawa, H.; Shibata, N.; Shibayama, K.; Arakawa, Y. A Preliminary Survey of Extended-Spectrum β-Lactamases (ESBLs) in Clinical Isolates of Klebsiella Pneumoniae and Escherichia coli in Japan. FEMS Microbiol. Lett. 2000, 184, 53–56. [Google Scholar] [CrossRef]
  98. Kruger, T.; Szabo, D.; Keddy, K.H.; Deeley, K.; Marsh, J.W.; Hujer, A.M.; Bonomo, R.A.; Paterson, D.L. Infections with Nontyphoidal Salmonella Species Producing TEM-63 or a Novel TEM Enzyme, TEM-131, in South Africa. Antimicrob. Agents Chemother. 2004, 48, 4263–4270. [Google Scholar] [CrossRef] [Green Version]
  99. de Oliveira Garcia, D.; Doi, Y.; Szabo, D.; Adams-Haduch, J.M.; Vaz, T.M.I.; Leite, D.; Padoveze, M.C.; Freire, M.P.; Silveira, F.P.; Paterson, D.L. Multiclonal Outbreak of Klebsiella Pneumoniae Producing Extended-Spectrum β-Lactamase CTX-M-2 and Novel Variant CTX-M-59 in a Neonatal Intensive Care Unit in Brazil. Antimicrob. Agents Chemother. 2008, 52, 1790–1793. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  100. Fernandes, S.A.; Camargo, C.H.; Francisco, G.R.; Bueno, M.F.C.; Garcia, D.O.; Doi, Y.; Casas, M.R.T. Prevalence of Extended-Spectrum β-Lactamases CTX-M-8 and CTX-M-2-Producing Salmonella Serotypes from Clinical and Nonhuman Isolates in Brazil. Microb. Drug Resist. 2017, 23, 580–589. [Google Scholar] [CrossRef] [PubMed]
  101. Pitout, J.D.D.; Hossain, A.; Hanson, N.D. Phenotypic and Molecular Detection of CTX-M-β-Lactamases Produced by Escherichia coli and Klebsiella Spp. J. Clin. Microbiol. 2004, 42, 5715–5721. [Google Scholar] [CrossRef] [Green Version]
  102. Cejas, D.; Fernández Canigia, L.; Quinteros, M.; Giovanakis, M.; Vay, C.; Lascialandare, S.; Mutti, D.; Pagniez, G.; Almuzara, M.; Gutkind, G.; et al. Plasmid-Encoded AmpC (pAmpC) in Enterobacteriaceae: Epidemiology of Microorganisms and Resistance Markers. Rev. Argent. Microbiol. 2012, 44, 182–186. [Google Scholar] [PubMed]
Table
Table 1. Occurrence of virulence factor-encoding genes in 112 uropathogenic E. coli (UPEC) isolates identified in the distinct E. coli phylogroups.
Table 1. Occurrence of virulence factor-encoding genes in 112 uropathogenic E. coli (UPEC) isolates identified in the distinct E. coli phylogroups.
Genes Investigated aNo. (%) of UPEC Isolates Identified in the Distinct Phylogroups
A
(n = 18)		B1
(n = 16)		B2
(n = 46)		C
(n = 2)		D
(n = 14)		E
(n = 2)		F
(n = 8)		G
(n = 4)		E. Clades
(n = 2)		Total
(n = 112)
Adhesins
fimH16 (88.9)16 (100)46 (100)2 (100)14 (100)2 (100)8 (100)4 (100)2 (100)110 (98.2)
ecpA11 (61.1)14 (87.5)36 (78.3)2 (100)12 (85.7)2 (100)6 (75)4 (100)087 (77.7)
papC1 (5.6)2 (12.5)18 (39.1)010 (71.4)02 (25)1 (25)1 (50)35 (31.3)
iha2 (11.1)1 (6.3)13 (28.3)08 (57.1)01 (12.5)1 (25)0 (0)26 (23.2)
papA1 (5.6)1 (6.3)10 (21.7)08 (57.1)03 (37.5)1 (25)1 (50)25 (22.3)
sfa/focDE0019 (41.3)00000019 (17)
afaBC1 (5.6)03 (6.5)00001 (25)05 (4.5)
hra00001 (7.1)01 (12.5)002 (1.8)
Invasin
ibeA0016 (34.8)0001 (12.5)0017 (15.2)
Toxins
usp0033 (71.7)01 (7.1)02 (25)0036 (32.1)
sat1 (5.6)1 (6.3)11 (23.9)09 (64.3)03 (37.5)1 (25)026 (23.2)
vat0024 (52.2)00001 (25)025 (22.3)
hlyA1 (5.6)08 (17.4)01 (7.1)000010 (8.9)
picU1 (5.6)06 (13)01 (7.1)01 (12.5)1 (25)010 (8.9)
hlyF03 (18.8)3 (6.5)1 (50)0001 (25)08 (7.1)
cnf1006 (13)0000006 (5.4)
cdt002 (4.3)0000002 (1.8)
Iron Acquisition
irp24 (22.2)3 (18.8)35 (76.1)1 (50)9 (64.3)03 (37.5)4 (100)059 (52.7)
sitA4 (22.2)5 (31.3)34 (73.9)2 (100)9 (64.3)03 (37.5)1 (25)058 (51.8)
iucD1 (5.6)7 (43.8)20 (43.5)1 (50)6 (42.9)04 (50)2 (50)041 (36.6)
ireA008 (17.4)01 (7.1)001 (25)010 (8.9)
iroN01 (6.3)4 (8.7)0000005 (4.5)
Serum Resistance
traT11 (61.1)13 (81.3)38 (82.6)2 (100)14 (100)2 (100)6 (75)4 (100)1 (50)91 (81.3)
ompT6 (33.3)7 (43.8)38 (82.6)1 (50)11 (78.6)1 (50)4 (50)2 (50)070 (62.5)
iss1 (5.6)2 (12.5)5 (10.9)2 (100)0000010 (8.9)
cva03 (18.8)1 (2.2)00001 (25)05 (4.5)
kpsMTII002 (4.3)01 (7.1)01 (12.5)004 (3.6)
Range of VFs0–112–93–176–84–123–42–115–112–30–17
Mean of VFs3.44.99.57.08.33.56.17.82.57.2
a The tsh and bmaE genes were not detected in any of the UPEC isolates studied. VFs: Virulence factor-encoding genes.
Table
Table 2. Serotype, E. coli phylogroup classification, and molecular characteristics of the hybrid uropathogenic/diarrheogenic E. coli (UPEC/DEC) isolates identified in this study.
Table 2. Serotype, E. coli phylogroup classification, and molecular characteristics of the hybrid uropathogenic/diarrheogenic E. coli (UPEC/DEC) isolates identified in this study.
Hybrid UPEC IdentificationSerotypePhylogroupDEC a MarkersExPEC Related Virulence
Factor-Encoding Genes
EPECEAEC
escNbfpBaatAaggR
34 O3:H2A--++fimH, ecpA, papA/papC, iha, afaBC, hlyA, sat, irp2, traT, ompT
69 O15:H6D--++fimH, ecpA, papA/papC, iha, hlyA, sat, irp2, traT, ompT, picU
85 O126:H10B1--++fimH, ecpA, irp2, traT, ompT, sitA, iucD
92 O145:H34B2+---fimH, ecpA, ibeA, traT, ompT
100 O90:H2D--++fimH, ecpA, papC, traT, ompT, sitA
a Diarrheagenic Escherichia coli (DEC) pathotypes: EPEC: enteropathogenic E. coli; EAEC: enteroaggregative E. coli. None of the UPEC isolates studied harbored the stx1 and/or stx2 genes.
Table
Table 3. Age, gender, type of urinary tract infection (UTI) and additional relevant clinical information from patients diagnosed with UTIs from which hybrid isolates of uropathogenic and diarrheagenic (UPEC/DEC) E. coli were obtained.
Table 3. Age, gender, type of urinary tract infection (UTI) and additional relevant clinical information from patients diagnosed with UTIs from which hybrid isolates of uropathogenic and diarrheagenic (UPEC/DEC) E. coli were obtained.
Hybrid UPEC/DEC
Identification		Type of
Hybrid a		Patient Data:
Age
(Years)		Gender bType of
UTI		Additional
Information
34UPEC/EAEC11FCystitisAutistic Spectrum Disorder
69UPEC/EAEC22F cPyelonephritisFever
85UPEC/EAEC1MCystitisPrunc Belly Syndrome
92UPEC/aEPEC84FCystitisDiabetes melitus, hypertension and coronary artery disease
100UPEC/EAEC40FCystitisRenal transplantation and use of immunosuppressants
a UPEC: uropathogenic E. coli, EAEC: enteroaggregative E. coli, and aEPEC: atypical enteropathogenic E. coli. b F: female and M: male. c Ten weeks of pregnancy.
Table
Table 4. Antimicrobial resistance of the uropathogenic E. coli (UPEC) isolates studied.
Table 4. Antimicrobial resistance of the uropathogenic E. coli (UPEC) isolates studied.
Antimicrobial Drugs TestedNo. (%) of UPEC Isolates Classified in Each Category:
SusceptibleIntermediate aResistant
β-lactams
Ampicillin (AMP)60 (53.6)052 (46.4)
Piperacillin/Tazobactam (PPT)109 (97.3)3 (2.7)0
Ampicillin/Sulbactam (ASB)95 (84.8)11 (9.8)6 (5.4)
Amoxicillin/clavulanic acid (AMC)93 (83)15 (13.4)4 (3.6)
Cephalothin (CFL)51 (45.5)40 (35.7)21 (18.8)
Cefuroxime (CRX)100 (89.3)012 (10.7)
Ceftazidime (CAZ)105 (93.8)1 (0.9)6 (5.4)
Cefotaxime (CTX)101 (90.2)011 (9.8)
Cefepime (CPM)100 (89.3)2 (1.8) a10 (8.9)
Ertapenem (ETP)112 (100)0 (0)0 (0)
Meropenem (MER)112 (100)0 (0)0 (0)
Aminoglycosides
Gentamicin (GEN)96 (85.7)11 (9.8)5 (4.5)
Amikacin (AMI)105 (93.8)7 (6.3)0
Quinolones/Fluoroquinolones
Nalidixic acid (NAL)69 (61.6)8 (7.1)35 (31.3)
Norfloxacin (NOR)87 (77.7)2 (1.8)23 (20.5)
Ciprofloxacin (CIP)63 (56.3)14 (12.5)35 (31.3)
Folic acid metabolism inhibitors
Trimethoprim/Sulfamethoxazole (SUT)73 (65.2)039 (34.8)
Nitrofuran
Nitrofurantoin (NIT)111 (99.1)1 (0.9)0
Phosphonic acid derivative
Fosfomycin (FOS)111 (99.1)01 (0.9)
a Or susceptible-dose dependent (SDD), if appropriate.
Table
Table 5. Phenotypic and molecular features of the eleven Extended Spectrum Beta-Lactamase (ESBL)-producing uropathogenic E. coli (UPEC) isolates identified in this study.
Table 5. Phenotypic and molecular features of the eleven Extended Spectrum Beta-Lactamase (ESBL)-producing uropathogenic E. coli (UPEC) isolates identified in this study.
UPEC
Identification		SerotypePhylogroupResistance Profile to β-Lactams:β-Lactamase-Encoding Genes
Penicillins a,dCephalosporins b,dCephalosporinase cAdditional Genes
1st Gen.2nd Gen.3rd Gen.4th Gen.
10 ONT:HNME. cladesAMP (R)CFL (R)CRX (R)CTX (R)CPM (R)blaCTX-M-8blaTEM-1
20 ONT:HNMAAMP (R)CFL (R)CRX (R)CAZ (R), CTX (R)CPM (R)blaCTX-M-15-
30 ONT:HNMAAMP (R)CFL (R)CRX (R)CAZ (I), CTX (R)CPM (R)blaCTX-M-15-
55 OR:H51B1AMP (R), PPT (I), AMC (R)CFL (R)CRX (R)CAZ (R), CTX (R)CPM (R)blaCTX-M-15, blaCMY-2blaTEM-1
60 ONT:HNME. cladesAMP (R), ASB (I), AMC (R)CFL (R)CRX (R)CAZ (R), CTX (R)CPM (R)blaCTX-M-15-
85 O126:H10B1AMP (R), ASB (R), AMC (I)CFL (R)CRX (R)CTX (R)CPM (R)blaCTX-M-8blaTEM-1
94 OR:H4B1AMP (R), AMC (R)CFL (R)CRX (R)CAZ (R), CTX (R)CPM (R)blaCTX-M-15, blaCMY-2-
109 O101:HNMAAMP (R), ASB (I)CFL (R)CRX (R)CTX (R)CPM (R)blaCTX-M-8blaTEM-1
112 ONT:H4AAMP (R), ASB (I), AMC (I)CFL (R)CRX (R)CAZ (R), CTX (R)CPM (R)blaCTX-M-15-
114 O25:H4B2AMP (R), ASB (I), AMC (I)CFL (R)CRX (R)CAZ (R), CTX (R)CPM (R)blaCTX-M-15blaTEM-1
120 OR:H6FAMP (R)CFL (R)CRX (R)CTX (R)CPM (I)blaCTX-M-15-
a Penicillins: ampicillin (AMP), Piperacillin/Tazobactam (PPT), ampicillin/sulbactam (ASB), amoxicillin/clavulanic acid (AMC). b Cephalosporins: cephalothin (CFL), cefuroxime (CRX), ceftazidime (CAZ), cefotaxime (CTX), and cefepime (CPM). c Cephalosporinase CTX-M-genes refer to groups. d Letters in parentheses indicate: resistant (R) or intermediate (I).
Table
Table 6. Resistance profile of hybrid uropathogenic/diarrheogenic E. coli (UPEC/DEC) isolates identified in the present study.
Table 6. Resistance profile of hybrid uropathogenic/diarrheogenic E. coli (UPEC/DEC) isolates identified in the present study.
Hybrid UPEC
Identification		Type of
Hybrid E. coli a		ESBL-Producing UPECClasses of Antimicrobial Drugs b,c:
β-LactamAminoglycosideQuinolone/FluoroquinoloneFolic Acid
Metabolism
Inhibitor
34UPEC/EAEC-AMP (R), ASB (R), AMC (I), CFL (I)---
69UPEC/EAEC-AMP (R), ASB (I), AMC (I)---
85UPEC/EAEC+AMP (R), ASB (R), AMC (I), CFL (R), CRX (R), CTX (R), CPM (R)GEN (I)NAL (I), CIP (R)SUT (R)
92UPEC/aEPEC-----
100UPEC/EAEC-AMP (R), CFL (I)GEN (I)NAL (I)SUT (R)
a UPEC: uropathogenic E. coli, EAEC: enteroaggregative E. coli, and aEPEC: atypical enteropathogenic E. coli. b Classes of antimicrobial drugs: β-Lactams: ampicillin (AMP), ampicillin/sulbactam (ASB), amoxicillin/clavulanic acid (AMC), cephalothin (CFL), cefuroxime (CRX), cefotaxime (CTX), cefepime (CPM); Aminoglycoside: gentamicin (GEN); Quinolones: nalidixic acid (NAL), ciprofloxacin (CIP); Folic acid metabolism inhibitors: trimethoprim/sulfamethoxazole (SUT). c Letters in parentheses indicate: resistant (R) or intermediate (I).
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Tanabe, R.H.S.; Dias, R.C.B.; Orsi, H.; de Lira, D.R.P.; Vieira, M.A.; dos Santos, L.F.; Ferreira, A.M.; Rall, V.L.M.; Mondelli, A.L.; Gomes, T.A.T.; et al. Characterization of Uropathogenic Escherichia coli Reveals Hybrid Isolates of Uropathogenic and Diarrheagenic (UPEC/DEC) E. coli. Microorganisms 2022, 10, 645. https://doi.org/10.3390/microorganisms10030645

AMA Style

Tanabe RHS, Dias RCB, Orsi H, de Lira DRP, Vieira MA, dos Santos LF, Ferreira AM, Rall VLM, Mondelli AL, Gomes TAT, et al. Characterization of Uropathogenic Escherichia coli Reveals Hybrid Isolates of Uropathogenic and Diarrheagenic (UPEC/DEC) E. coli. Microorganisms. 2022; 10(3):645. https://doi.org/10.3390/microorganisms10030645

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Tanabe, Rodrigo H. S., Regiane C. B. Dias, Henrique Orsi, Daiany R. P. de Lira, Melissa A. Vieira, Luís F. dos Santos, Adriano M. Ferreira, Vera L. M. Rall, Alessandro L. Mondelli, Tânia A. T. Gomes, and et al. 2022. "Characterization of Uropathogenic Escherichia coli Reveals Hybrid Isolates of Uropathogenic and Diarrheagenic (UPEC/DEC) E. coli" Microorganisms 10, no. 3: 645. https://doi.org/10.3390/microorganisms10030645

APA Style

Tanabe, R. H. S., Dias, R. C. B., Orsi, H., de Lira, D. R. P., Vieira, M. A., dos Santos, L. F., Ferreira, A. M., Rall, V. L. M., Mondelli, A. L., Gomes, T. A. T., Camargo, C. H., & Hernandes, R. T. (2022). Characterization of Uropathogenic Escherichia coli Reveals Hybrid Isolates of Uropathogenic and Diarrheagenic (UPEC/DEC) E. coli. Microorganisms, 10(3), 645. https://doi.org/10.3390/microorganisms10030645

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