Clonal Relatedness, Phylotyping and Antimicrobial Susceptibility of Extended- spectrum-beta-lactamase Producing Uropathogenic Escherichia coli Isolates from Outpatients and Inpatients

2020 ◽  
Vol 20 (5) ◽  
pp. 659-666
Author(s):  
Negar Azimzadeh ◽  
Abdollah Derakhshandeh ◽  
Mohammad Motamedifar ◽  
Zahra Naziri

Objectives: Antibiotic resistance, phylogenetic groups and Pulsed-Field Gel Electrophoresis (PFGE) patterns were evaluated in urinary tract infection (UTI) Escherichia coli (E. coli) isolates from outpatients and inpatients. Methods: In this study, antibiotic resistance to E. coli isolated from non-hospitalized and hospitalized patients (153 outpatients and 147 inpatients ) was evaluated in Shiraz County, Iran. Phylogenetic groups and Pulse Field Gel Electrophoresis (PFGE) patterns of 143 ESBLs-producing E. coli were also assessed. Results: The prevalence of ESBL-producing E. coli was shown to be 46.4% and 49% in the outpatient and inpatient UTI E. coli isolates, respectively. Most ESBL-producers were detected on patients hospitalized in clinical surgery units (66.7%) and intensive care units (62.5%). Phylogenetic group D was the dominant group in both the outpatient and inpatient isolates (67.6% and 61.1%, respectively) and also in internal, clinical surgery and ICU units. PFGE results showed more relatedness (>80% similarity) among inpatient isolates. PFGE analysis of 49 ESBL-producing inpatient E.coli in hospital units revealed 17 different pulsotypes, consisting of 11 clones and 6 single patterns. There were no clonal patterns in outpatient isolates, and similarity among the outpatient isolates and also between inpatient and outpatient isolates was less than 80% (75% and 66%, respectively). Conclusions: The results showed extreme genomic diversity among the ESBL-producing E. coli isolates in terms of the community and multiclonal dissemination of ESBL-producing E. coli isolated from hospital units.

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 262
Author(s):  
Isabel Carvalho ◽  
Nadia Safia Chenouf ◽  
Rita Cunha ◽  
Carla Martins ◽  
Paulo Pimenta ◽  
...  

The aim of the study was to analyze the mechanisms of resistance in extended-spectrum beta-lactamase (ESBL)- and acquired AmpC (qAmpC)-producing Escherichia coli isolates from healthy and sick cats in Portugal. A total of 141 rectal swabs recovered from 98 sick and 43 healthy cats were processed for cefotaxime-resistant (CTXR) E. coli recovery (in MacConkey agar supplemented with 2 µg/mL cefotaxime). The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) method was used for E. coli identification and antimicrobial susceptibility was performed by a disk diffusion test. The presence of resistance/virulence genes was tested by PCR sequencing. The phylogenetic typing and multilocus sequence typing (MLST) were determined by specific PCR sequencing. CTXRE. coli isolates were detected in seven sick and six healthy cats (7.1% and 13.9%, respectively). Based on the synergy tests, 11 of 13 CTXRE. coli isolates (one/sample) were ESBL-producers (ESBL total rate: 7.8%) carrying the following ESBL genes: blaCTX-M-1 (n = 3), blaCTX-M-15 (n = 3), blaCTX-M-55 (n = 2), blaCTX-M-27 (n = 2) and blaCTX-M-9 (n = 1). Six different sequence types were identified among ESBL-producers (sequence type/associated ESBLs): ST847/CTX-M-9, CTX-M-27, CTX-M-1; ST10/CTX-M-15, CTX-M-27; ST6448/CTX-M-15, CTX-M-55; ST429/CTX-M-15; ST101/CTX-M-1 and ST40/CTX-M-1. Three of the CTXR isolates were CMY-2-producers (qAmpC rate: 2.1%); two of them were ESBL-positive and one ESBL-negative. These isolates were typed as ST429 and ST6448 and were obtained in healthy or sick cats. The phylogenetic groups A/B1/D/clade 1 were detected among ESBL- and qAmpC-producing isolates. Cats are carriers of qAmpC (CMY-2)- and ESBL-producing E. coli isolates (mostly of variants of CTX-M group 1) of diverse clonal lineages, which might represent a public health problem due to the proximity of cats with humans regarding a One Health perspective.


2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Mourouge Saadi Alwash ◽  
Hawraa Mohammed Al-Rafyai

Surface water contamination remains a major worldwide public health concern and may contribute to the dissemination of antibiotic-resistant bacteria. The Al-Hillah River in the city of Babylon Province, Iraq, diverts flows from the Euphrates River. Because of its importance in irrigation and population density, it faces several forced and unforced changes due to anthropogenic activities. To evaluate water quality, water samples were collected from three sites with different anthropogenic pressures along the Al-Hillah River. These samples were subjected to bacteriological analyses, i.e., total coliforms, Escherichia coli, and faecal enterococci. The phylogenetic groups of the E. coli isolates (n = 61) were typed by rapid PCR-based analyses. Representatives of each isolate were tested phenotypically for resistance to six classes of antibiotics and characterized according to their phylogenetic groups. The results demonstrated the highest resistance levels were to β-lactam antibiotics, followed by fosfomycin and aminoglycosides. Escherichia coli isolates belonging to phylogenetic groups A and B2 were the most common and were characterized by a higher prevalence of antibiotic resistance. This study is important for understanding the current conditions of the Al-Hillah River, as the data reveal a high prevalence of multiresistance among E. coli isolates circulating at the three sampling sites.


2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Susan Mosquito ◽  
Maria J. Pons ◽  
Maribel Riveros ◽  
Joaquim Ruiz ◽  
Theresa J. Ochoa

Conventionally, inEscherichia coli, phylogenetic groups A and B1 are associated with commensal strains while B2 and D are associated with extraintestinal strains. The aim of this study was to evaluate diarrheagenic (DEC) and commensalE. coliphylogeny and its association with antibiotic resistance and clinical characteristics of the diarrheal episode. Phylogenetic groups and antibiotic resistance of 369E. colistrains (commensal strains and DEC from children with or without diarrhea) isolated from Peruvian children <1 year of age were determined by a Clermont triplex PCR and Kirby-Bauer method, respectively. The distribution of the 369E. colistrains among the 4 phylogenetic groups was A (40%), D (31%), B1 (21%), and B2 (8%). DEC-control strains were more associated with group A while DEC-diarrhea strains were more associated with group D(P<0.05). There was a tendency(P=0.06)for higher proportion of persistent diarrhea (≥14 days) among severe groups (B2 and D) in comparison with nonsevere groups (A and B1). Strains belonging to group D presented significantly higher percentages of multidrug resistance than the rest of the groups(P>0.01). In summary, DEC-diarrhea strains were more associated with group D than strains from healthy controls.


Water ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 3449
Author(s):  
Cristina-Mirabela Gaşpar ◽  
Ludovic Toma Cziszter ◽  
Cristian Florin Lăzărescu ◽  
Ioan Ţibru ◽  
Marius Pentea ◽  
...  

This study aimed to compare the antibiotic resistance levels of the indicator bacteria Escherichia coli in wastewater samples collected from two hospitals and two urban communities. Antimicrobial susceptibility testing was performed on 81 E. coli isolates (47 from hospitals and 34 from communities) using the disc diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology. Ten antibiotics from nine different classes were chosen. The strains isolated from the community wastewater, compared to those from the hospital wastewater, were not resistant to gentamicin (p = 0.03), but they showed a significantly higher susceptibility—increased exposure to ceftazidime (p = 0.001). Multidrug resistance was observed in 85.11% of the hospital wastewater isolates and 73.53% of the community isolates (p > 0.05). The frequency of the presumed carbapenemase-producing E. coli was higher among the community isolates (76.47% compared to 68.09%) (p > 0.05), whereas the frequency of the presumed extended-spectrum beta-lactamase (ESBL)-producing E. coli was higher among the hospital isolates (21.28% compared to 5.88%) (p > 0.05). The antibiotic resistance rates were high in both the hospital and community wastewaters, with very few significant differences between them, so the community outlet might be a source of resistant bacteria that is at least as important as the well-recognised hospitals.


2017 ◽  
Vol 11 (01) ◽  
pp. 51-57 ◽  
Author(s):  
Yandag Munkhdelger ◽  
Nyamaa Gunregjav ◽  
Altantsetseg Dorjpurev ◽  
Nishi Juniichiro ◽  
Jav Sarantuya

Introduction: The severity of urinary tract infection (UTI) produced by uropathogenic Escherichia coli (UPEC) is due to the expression of a wide spectrum of virulence genes. E. coli strains were divided into four phylogenetic groups (A, B1, B2 and D) based on their virulence genes. The present study aimed to assess the relationship between virulence genes, phylogenetic groups, and antibiotic resistance of UPEC. Methodology: A total of 148 E. coli were tested for antimicrobial resistance against 10 drugs using the disk diffusion method. The isolates were screened by polymerase chain reaction (PCR) for detection of virulence genes and categorized into the four major phylogenetic groups. Results: Phylogenetic group B2 was predominant (33.8%), followed by D (28.4%), A (19.6), and B1 (18.2%). A higher prevalence of fimH (89.9%), fyuA (70.3%), traT (66.2%), iutA (62.2%), kpsMTII (58.8%), and aer (56.1%) genes were found in UPEC, indicating a putative role of adhesins, iron acquisition systems, and protectins that are main cause of UTIs. The most common antibiotic resistance was to cephalotin (85.1%), ampicillin (78.4%) and the least to nitrofurantoin (5.4%) and imipenem (2%). In total, 93.9% of isolates were multidrug resistant (MDR). Conclusions: This study showed that group B2 and D were the predominant phylogenetic groups and virulence-associated genes were mostly distributed in these groups. The virulence genes encoding components of adhesins, iron acquisition systems, and protectins were highly prevalent among antibiotic-resistant UPEC. Although the majority of strains are MDR, nitrofurantoin is the drug of choice for treatment of UTI patients in Ulaanbaatar.


2021 ◽  
Vol 19 (3) ◽  
pp. 349-362
Author(s):  
Prayuth Saekhow ◽  
◽  
Chayaphon Sriphannam ◽  
◽  

We investigated the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains in dairy farm wastewater in Chiang Mai, Thailand. We analyzed wastewater samples collected from 150 dairy farms and found that 88.7% of the farms (n = 133) were positive for ESBL-producing E. coli. Multiplex polymerase chain reaction (PCR) amplification was performed to characterize the presence of bla CTX-M, bla TEM, and blaSHV in ESBL-producing isolates. blaCTX-M was found in all isolates (n = 133), followed by blaTEM (80/133, 60.2%), whereas blaSHV was not detected in any isolate. blaCTX-M and blaTEM were present in 60.2% (80/133) of the isolates, and 39.8% (53/133) isolates carried bla CTX-M alone. Subgroup analysis showed that CTX-M-1 was the most prevalent subgroup among the isolates (129/133, 97.0%), followed by CTX-M-8 (2/133, 1.5%) and CTX-M-9 (2/133, 1.5%). The distribution of the phylogenetic groups was as follows: group A (100/133, 75.2%), followed by B1 (14/133, 10.5%), D (6/133, 4.5%), F (6/133, 4.5%), B2 (4/133, 3.0%), and E (3/133, 2.3%). Based on enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and dendrogram analysis, 24 isolates were classified into clades I (n = 21), II (n =1), and III (n =2). Minor genetic differences were found in all clade I isolates. Our data suggest that the circulating of ESBL-producing E. coli carried at least one bla gene strain distributed in dairy farm wastewater in Chiang Mai.


Antibiotics ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 161 ◽  
Author(s):  
Saskia-Camille Flament-Simon ◽  
Marie-Hélène Nicolas-Chanoine ◽  
Vanesa García ◽  
Marion Duprilot ◽  
Noémie Mayer ◽  
...  

Escherichia coli is the main pathogen responsible for extraintestinal infections. A total of 196 clinical E. coli consecutively isolated during 2016 in Spain (100 from Lucus Augusti hospital in Lugo) and France (96 from Beaujon hospital in Clichy) were characterized. Phylogroups, clonotypes, sequence types (STs), O:H serotypes, virulence factor (VF)-encoding genes and antibiotic resistance were determined. Approximately 10% of the infections were caused by ST131 isolates in both hospitals and approximately 60% of these infections were caused by isolates belonging to only 10 STs (ST10, ST12, ST58, ST69, ST73, ST88, ST95, ST127, ST131, ST141). ST88 isolates were frequent, especially in Spain, while ST141 isolates significantly predominated in France. The 23 ST131 isolates displayed four clonotypes: CH40-30, CH40-41, CH40-22 and CH40-298. Only 13 (6.6%) isolates were carriers of extended-spectrum beta-lactamase (ESBL) enzymes. However, 37.2% of the isolates were multidrug-resistant (MDR). Approximately 40% of the MDR isolates belonged to only four of the dominant clones (B2-CH40-30-ST131, B2-CH40-41-ST131, C-CH4-39-ST88 and D-CH35-27-ST69). Among the remaining MDR isolates, two isolates belonged to B2-CH14-64-ST1193, i.e., the new global emergent MDR clone. Moreover, a hybrid extraintestinal pathogenic E.coli (ExPEC)/enteroaggregative isolate belonging to the A-CH11-54-ST10 clone was identified.


2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Sahar Besharati Zadeh ◽  
Pegah Shakib ◽  
Mohammad Reza Zolfaghari ◽  
Ahmad Farajzadeh Sheikh

Background: A major problem in the treatment of the infectious diseases healthcare centers is extended-spectrum beta-lactamase (ESBL)-producing bacteria. Objectives: The aim of present study was to identify the antibiotic sensitivity pattern and prevalence of the blaCTX, blaTEM, and blaSHV genes in Escherichia coli and Klebsiella pneumoniae strains. Methods: In this study, E. coli and K. pneumoniae specimens were collected in Shushtar hospitals, Khuzestan (southwest Iran), from March to October 2015. Sensitivity antibiotic pattern performed by disc diffusion method. Double disc synergy test (DDST) done for identifying ESBLs isolates and PCR for blaTEM, blaSHV, and blaCTX-M genes. Results: One hundred E. coli and 30 K. pneumoniae isolates were collected from different specimens. The highest rates of antibiotic resistance related to cefotaxime and aztreonam in E. coli and K. pneumoniae. ESBL-harboring K. pneumoniae and E. coli were 13.5 and 28%, respectively. Overall, bla TEM was the most prevalent ESBL gene. Conclusions: In this study, the rate of antibiotic resistance was high, and due to the carrying of coding genes on mobile genetic elements and the ability of these elements to carry genes that create resistance to other antibiotic families, identification and isolation of these isolates are essential to find effective antibiotics and eliminate the infection.


Author(s):  
Hossein Norouzian ◽  
Mohammad Katouli ◽  
Nader Shahrokhi ◽  
Sharam Sabeti ◽  
Mohammad Pooya ◽  
...  

Background and Objectives: B2 and D have been mentioned as the most common phylogenetic groups among uropatho- genic Escherichia coli. However, there is still controversy about the importance of these phylo-groups. This study was con- ducted to investigate the probable relation between these groups and antibiotic resistance patterns of E. coli isolates derived from urine and feces of the patients with acute or recurrent UTI. Materials and Methods: 10 isolates were recovered from urine and feces samples of patients with different phases of UTI in whom E. coli was causative pathogen. Biochemical fingerprinting was performed to classify the isolates and select their appropriate representatives. Phylogenetic grouping was performed using multiplex PCR, and antibiotic resistance was deter- mined by disk diffusion method. Results: Five-hundred-sixty E. coli isolates were derived from 56 UTI patients (27 acute, 29 recurrent). Among them, 261 isolates were selected using biochemical fingerprinting. All the isolates were sensitive to imipenem and nitrofurantoin. Com- pared to other phylo-groups, the isolates in group D showed considerably different frequencies in acute vs. recurrent phase of UTI, in urine vs. stool samples, in males vs. females, and in- vs. out-patients. They were more resistant to the antibiotics (except norfloxacin), and in contrast to others, this was seen more in acute UTI, especially in urine samples. Multi-drug resistance pattern was also meaningfully higher in group D. Conclusion: Although phylo-groups B2 and D of E. coli bacteria are more responsible for UTI, group D isolates seem to be more resistant and probably more virulent, even than the ones from group B2.


2021 ◽  
Vol 14 (2) ◽  
Author(s):  
Mostafa Boroumand Boroumand ◽  
Mohsen Naghmachi ◽  
Mohammad Amin Ghatee

Background: Many bacteria can cause urinary tract infections (UTIs), among which Escherichia coli is the most common causative agent. E. coli strains are divided into eight phylogenetic groups based on the new Quadroplex-PCR method, which are different in terms of patterns of resistance to antibiotics, virulence, and environmental characteristics. Objectives: This study aimed to determine the phylogenetic groups and the prevalence of drug resistance genes in E. coli strains causing UTIs. Methods: In this descriptive cross-sectional study, 129 E. coli isolates obtained from the culture of patients with UTIs were evaluated for phylogenetic groups using the new method of Clermont et al. The identification of phylogenetic groups and antibiotic resistance genes was performed using the multiplex polymerase chain reaction (PCR) method. Results: In this study, concerning the distribution of phylogenetic groups among E. coli isolates, the phylogenetic group B2 (36.4%) was the most common phylogenetic group, followed by phylogroups C (13.2%), clade I (10.1%), D (9.3%), and A (3.1%) while groups B1 and F were not observed in any of the isolates, and 20.2% had an unknown state. Also, out of 129 E. coli isolates, the total frequency of tetA, tetB, sul1, sul2, CITM, DfrA, and qnr resistance genes was 59.7%, 66.7, 69, 62, 30.2, 23.3, and 20.2%, respectively. In this study, there was a significant relationship between antibiotics (P = 0.026), cefotaxime (P = 0.003), and nalidixic acid (P = 0.044) and E. coli phylogenetic groups. No significant relationship was observed between E. coli phylogenetic groups and antibiotic resistance genes. Conclusions: The results of this study showed that strains belonging to group B2 had the highest prevalence among other phylogroups, and also, the frequency of antibiotic resistance genes and drug-resistant isolates had a higher prevalence in this phylogroup. These results show that phylogroup B2 has a more effective role in causing urinary tract infections compared to other phylogroups, and this phylogroup can be considered a genetic reservoir of antibiotic resistance.


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