Molecular characterization of virulence genes and antibiotic resistance among fecal Escherichia coli isolated from surface water of Wadi Shueib-Jordan

Objective. Contamination of surface water with pathogenic organism is highly dangerous for people who used it for drinking or for domestic activities. Detection of Escherichia coli in water can be used as a general important indicator for fecal contamination .This study investigated the occurrence of fecal E. coli and two important toxigenic types of E. coli isolates and their antimicrobial resistance in water samples collected from a surface water source ( Wadi Shueib) in Jordan . Methods. A total of 51 water samples were collected from three different locations of wadi shueib over a three month, July through September, 2016. For each sample, 200 ml of water was collected in sterilized containers. All samples which were positive for fecal E. coli were subcultured on Eosin methylelne blue agar and incubated at 37 ºC for 24 hours. E. coli isolates were identified by API- 20 E test , and all isolates were tested for antimicrobial susceptibility and for the presence of virulence genes of ETEC and EHEC. Results. A total of 46 /51 (90%) of water samples were contaminated with fecal E. coli. These 46 E. coli isolates were resistant in the range between 4%-76% to commonly used antibiotics in the treatment of urinary tract infection in Jordan. Multidrug resistant isolates to at least three antibiotics accounted for 17/46 (37%) of the isolates. Out of 46 fecal E. coli isolates, 4(8.7%) were enterotoxigenic E. coli (ETEC) and 2 (4.3%) were enterohemorrhagic E. coli (EHEC) as detected using PCR. Conclusion. This study indicated that the surface running water of Wadi Shueib is contaminated with potential enteropathogenic E. coli , and it can be a source of causing diarrheal diseases . Fecal E. coli isolates from water also showed high level of antibiotic resistance patterns comparable to human E. coli isolates in Jordan.

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Characterisation of multidrug resistant Escherichia coli isolated from two commercial lettuce and spinach supply chains

Journal of Food Protection ◽

10.4315/jfp-21-125 ◽

2021 ◽

Author(s):

Muneiwa T. Ratshilingano ◽

Erika Margarete du Plessis ◽

Stacey Duvenage ◽

Lise Korsten

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Irrigation Water ◽

Virulence Genes ◽

Disease Outbreaks ◽

Virulence Gene ◽

Multidrug Resistant ◽

Water Sources ◽

Point Of Sale ◽

E Coli

ABSTRACT Leafy green vegetables have increasingly been reported as a reservoir of multidrug-resistant pathogenic Enterobacteriaceae; with Shiga toxin- producing Escherichia coli frequently implicated in disease outbreaks worldwide. This study aimed to determine the presence and characteristics of antibiotic resistance, diarrheagenic virulence genes and phylogenetic groupings of E. coli isolates (n=51) from commercially produced lettuce and spinach from the farm, through processing and at the point of sale. Multidrug resistance was observed in 33 of the 51 E. coli isolates (64.7%); with 35.7% (n=10/28) being generic and 100% (n=23/23) Extended Spectrum β-lactamase/AmpC- producing. Resistance of E. coli isolates was observed against neomycin (100%; n=51/51), ampicillin (70.6%; n=36/51), amoxycilin (68.6%; n=35/51), tetracycline (45%; n=23/51), trimethoprim/ sulfamethoxazole (43%; n=22/51), chloramphenicol (25.5%; n=13/51), augmentin (11.8%; n=6/51) and gentamicin (7.8%; n=4/51); with 100% (n=51/51) susceptibility to imipenem. Virulence gene eae was detected in two E. coli isolates from irrigation water sources only, while none of the other virulence genes tested for were detected. Most of the E. coli strains belonged to phylogenetic group B2 (25.5%; n=13), B1 (19.6%; n=10) and A (17.6%; n=9); with D (5.9%; n=3) less distributed. Although diarrheagenic E. coli were not detected, antibiotic resistance in E. coli prevalent in the supply chain was evident. Additionally, a clear link between E. coli isolates from irrigation water sources and leafy green vegetables through DNA fingerprinting was established which indicates the potential transfer of E. coli from irrigation water to minimally processed leafy green vegetables.

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Diversity of P1 phage-like elements in multidrug resistant Escherichia coli

Scientific Reports ◽

10.1038/s41598-019-54895-4 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 12

Author(s):

Carola Venturini ◽

Tiziana Zingali ◽

Ethan R. Wyrsch ◽

Bethany Bowring ◽

Jonathan Iredell ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Multidrug Resistance ◽

Clinical Isolate ◽

Insertion Sequence ◽

Virulence Genes ◽

Multidrug Resistant ◽

Antibiotic Resistant ◽

E Coli ◽

Resistance Markers

AbstractThe spread of multidrug resistance via mobile genetic elements is a major clinical and veterinary concern. Pathogenic Escherichia coli harbour antibiotic resistance and virulence genes mainly on plasmids, but also bacteriophages and hybrid phage-like plasmids. In this study, the genomes of three E. coli phage-like plasmids, pJIE250-3 from a human E. coli clinical isolate, pSvP1 from a porcine ETEC O157 isolate, and pTZ20_1P from a porcine commensal E. coli, were sequenced (PacBio RSII), annotated and compared. All three elements are coliphage P1 variants, each with unique adaptations. pJIE250-3 is a P1-derivative that has lost lytic functions and contains no accessory genes. In pTZ20_1P and pSvP1, a core P1-like genome is associated with insertion sequence-mediated acquisition of plasmid modules encoding multidrug resistance and virulence, respectively. The transfer ability of pTZ20_1P, carrying antibiotic resistance markers, was also tested and, although this element was not able to transfer by conjugation, it was able to lysogenize a commensal E. coli strain with consequent transfer of resistance. The incidence of P1-like plasmids (~7%) in our E. coli collections correlated well with that in public databases. This study highlights the need to investigate the contribution of phage-like plasmids to the successful spread of antibiotic resistant pathotypes.

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Presence of β-Lactamase-producing Enterobacterales and Salmonella Isolates in Marine Mammals

International Journal of Molecular Sciences ◽

10.3390/ijms22115905 ◽

2021 ◽

Vol 22 (11) ◽

pp. 5905

Author(s):

Olivia M. Grünzweil ◽

Lauren Palmer ◽

Adriana Cabal ◽

Michael P. Szostak ◽

Werner Ruppitsch ◽

...

Keyword(s):

Escherichia Coli ◽

Marine Mammals ◽

Virulence Genes ◽

Multidrug Resistant ◽

Healthcare Sector ◽

Whole Genome ◽

E Coli ◽

High Prevalence ◽

Lactamase Gene ◽

Sequence Types

Marine mammals have been described as sentinels of the health of marine ecosystems. Therefore, the aim of this study was to investigate (i) the presence of extended-spectrum β-lactamase (ESBL)- and AmpC-producing Enterobacterales, which comprise several bacterial families important to the healthcare sector, as well as (ii) the presence of Salmonella in these coastal animals. The antimicrobial resistance pheno- and genotypes, as well as biocide susceptibility of Enterobacterales isolated from stranded marine mammals, were determined prior to their rehabilitation. All E. coli isolates (n = 27) were screened for virulence genes via DNA-based microarray, and twelve selected E. coli isolates were analyzed by whole-genome sequencing. Seventy-one percent of the Enterobacterales isolates exhibited a multidrug-resistant (MDR) pheno- and genotype. The gene blaCMY (n = 51) was the predominant β-lactamase gene. In addition, blaTEM-1 (n = 38), blaSHV-33 (n = 8), blaCTX-M-15 (n = 7), blaOXA-1 (n = 7), blaSHV-11 (n = 3), and blaDHA-1 (n = 2) were detected. The most prevalent non-β-lactamase genes were sul2 (n = 38), strA (n = 34), strB (n = 34), and tet(A) (n = 34). Escherichia coli isolates belonging to the pandemic sequence types (STs) ST38, ST167, and ST648 were identified. Among Salmonella isolates (n = 18), S. Havana was the most prevalent serotype. The present study revealed a high prevalence of MDR bacteria and the presence of pandemic high-risk clones, both of which are indicators of anthropogenic antimicrobial pollution, in marine mammals.

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Assessing the population dynamics of Escherichia coli in a metropolitan river after an extreme flood event

Journal of Water and Health ◽

10.2166/wh.2016.285 ◽

2016 ◽

Vol 15 (2) ◽

pp. 196-208 ◽

Cited By ~ 1

Author(s):

Nicole M. Masters ◽

Aaron Wiegand ◽

Jasmin M. Thompson ◽

Tara L. Vollmerhausen ◽

Eva Hatje ◽

...

Keyword(s):

Escherichia Coli ◽

Population Dynamics ◽

Antibiotic Resistance ◽

Virulence Genes ◽

Wastewater Treatment Plants ◽

Vero Cells ◽

Flood Event ◽

Typing Method ◽

E Coli ◽

Extreme Flood

We investigated Escherichia coli populations in a metropolitan river after an extreme flood event. Between nine and 15 of the 23 selected sites along the river were sampled fortnightly over three rounds. In all, 307 E. coli were typed using the PhP typing method and were grouped into common (C) or single (S) biochemical phenotypes (BPTs). A representative from each of the 31 identified C-BPTs was tested for 58 virulence genes (VGs) associated with intestinal and extra-intestinal E. coli, resistance to 22 antibiotics, production of biofilm and cytotoxicity to Vero cells. The number of E. coli in the first sampling round was significantly (P < 0.01) higher than subsequent rounds, whereas the number of VGs was significantly (P < 0.05) higher in isolates from the last sampling round when compared to previous rounds. Comparison of the C-BPTs with an existing database from wastewater treatment plants (WWTPs) in the same catchment showed that 40.6% of the river isolates were identical to the WWTP isolates. The relatively high number of VGs and antibiotic resistance among the C-BPTs suggests possessing and retaining these genes may provide niche advantages for those naturalised and/or persistent E. coli populations which may pose a health risk to the community.

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The population genomics of increased virulence and antibiotic resistance in human commensal Escherichia coli over 30 years in France

10.1101/2021.06.24.449745 ◽

2021 ◽

Author(s):

Julie Marin ◽

Olivier Clermont ◽

Guilhem Royer ◽

Melanie Mercier-Darty ◽

Jean-Winoc Decousser ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

High Risk ◽

Virulence Genes ◽

Population Genomics ◽

E Coli ◽

O Antigen ◽

Similar Region ◽

Intestinal Infections ◽

Evolution Of Virulence

Escherichia coli is a commensal species of the lower intestine, but also a major pathogen causing intestinal and extra-intestinal infections. Most studies on genomic evolution of E. coli used isolates from infections, and/or focused on antibiotic resistance, but neglected the evolution of virulence. Here instead, we whole-genome sequenced a collection of 436 E. coli isolated from fecal samples of healthy adult volunteers in France between 1980 and 2010. These isolates were distributed among 159 sequence types (STs), the five most frequent being ST10 (15.6%), ST73 (5.5%) and ST95 (4.8%), ST69 (3.7%) and ST59 (3.7%), and 230 O:H serotypes. ST and serotype diversity increased over time. Comparison with 912 E. coli bacteremia isolates from similar region and time showed a greater diversity in commensal isolates. The O1, O2, O6 and O25-groups used in bioconjugate O-antigen vaccine were found in only 63% of the four main STs associated with a high risk of bacteremia (ST69, ST73, ST95 and ST131). In commensals, STs associated with a high risk of bacteremia increased in frequency. Both extra-intestinal virulence-associated genes and resistance to antibiotics increased in frequency. Evolution of virulence genes was driven by both clonal expansion of STs with more virulence genes, and increases in frequency within STs, whereas the evolution of resistance was dominated by increases in frequency within STs. This study provides a unique picture of the phylogenomic evolution of E. coli in its human commensal habitat over a 30-year period and suggests that the efficacy of O-antigen vaccines would be threatened by serotype replacement.

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Antibiotic Resistance Profile of Commensal Escherichia coli Isolated from Broiler Chickens in Qatar

Journal of Food Protection ◽

10.4315/0362-028x.jfp-17-191 ◽

2017 ◽

Vol 81 (2) ◽

pp. 302-307 ◽

Cited By ~ 13

Author(s):

Nahla O. Eltai ◽

Elmoubasher A. Abdfarag ◽

Hamad Al-Romaihi ◽

Eman Wehedy ◽

Mahmoud H. Mahmoud ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Broiler Chickens ◽

Susceptibility Testing ◽

Multidrug Resistant ◽

Test Method ◽

Health Concern ◽

E Coli ◽

Extended Spectrum ◽

Stewardship Program

ABSTRACT Antibiotic resistance (AR) is a growing public health concern worldwide, and it is a top health challenge in the 21st century. AR among Enterobacteriaceae is rapidly increasing, especially in third-generation cephalosporins and carbapenems. Further, strains carrying mobilized colistin resistance (mcr) genes 1 and 2 have been isolated from humans, food-producing animals, and the environment. The uncontrolled use of antibiotics in food-producing animals is a major factor in the generation and spread of AR. No studies have been done to evaluate AR in the veterinary sector of Qatar. This study aimed at establishing primary baseline data for the prevalence of AR among food-producing animals in Qatar. Fecal samples (172) were obtained from two broiler farms and one live bird market in Qatar, and 90 commensal Escherichia coli bacteria were isolated and subjected to susceptibility testing against 16 clinically relevant antibiotics by using the E-test method. The results found that 81 (90%) of 90 isolates were resistant to at least one antibiotic, 14 (15.5%) of 90 isolates were colistin resistant, 2 (2.2%) of 90 isolates were extended-spectrum β-lactamase producers, and 2 (2.2%) of 90 isolates were multidrug resistant to four antibiotic classes. Extended-spectrum β-lactamase–producing E. coli and colistin-resistant isolates were confirmed by using double-disc susceptibility testing and PCR, respectively. Such a high prevalence of antibiotic-resistant E. coli could be the result of a long application of antibiotic treatment, and it is an indicator of the antibiotic load in food-producing animals in Qatar. Pathogens carrying AR can be easily transmitted to humans through consumption of undercooked food or noncompliance with hygiene practices, mandating prompt development and implementation of a stewardship program to control and monitor the use of antibiotics in the community and agriculture.

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Virulence Characteristics and Antibiotic Resistance Profiles of Shiga Toxin-Producing Escherichia coli Isolates from Diverse Sources

Antibiotics ◽

10.3390/antibiotics9090587 ◽

2020 ◽

Vol 9 (9) ◽

pp. 587

Author(s):

Momna Rubab ◽

Deog-Hwan Oh

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Shiga Toxin ◽

Multidrug Resistant ◽

Control Measures ◽

Diffusion Method ◽

Shiga Toxins ◽

E Coli ◽

And Control ◽

High Degree

Shiga toxin-producing Escherichia coli (STEC) is an enteric pathogen that causes several gastrointestinal ailments in humans across the world. STEC’s ability to cause ailment is attributed to the presence of a broad range of known and putative virulence factors (VFs) including those that encode Shiga toxins. A total of 51 E. coli strains belonging to serogroups O26, O45, O103, O104, O113, O121, O145, and O157 were tested for the presence of nine VFs via PCR and for their susceptibility to 17 frequently used antibiotics using the disc diffusion method. The isolates belonged to eight different serotypes, including eight O serogroups and 12 H types. The frequency of the presence of key VFs were stx1 (76.47%), stx2 (86.27%), eae (100%), ehxA (98.03%), nleA (100%), ureC (94.11%), iha (96.07%), subA (9.80%), and saa (94.11%) in the E. coli strains. All E. coli strains carried seven or more distinct VFs and, among these, four isolates harbored all tested VFs. In addition, all E. coli strains had a high degree of antibiotic resistance and were multidrug resistant (MDR). These results show a high incidence frequency of VFs and heterogeneity of VFs and MDR profiles of E. coli strains. Moreover, half of the E. coli isolates (74.5%) were resistant to > 9 classes of antibiotics (more than 50% of the tested antibiotics). Thus, our findings highlight the importance of appropriate epidemiological and microbiological surveillance and control measures to prevent STEC disease in humans worldwide.

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Virulence factors analysis and antibiotic resistance of uropathogenic Escherichia coli isolated from patients in northeast of Iran

Iranian Journal of Microbiology ◽

10.18502/ijm.v12i3.3240 ◽

2020 ◽

Author(s):

Mahdis Ghavidel ◽

Tahere Gholamhosseini-Moghadam ◽

Kimiya Nourian ◽

Kiarash Ghazvini

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Virulence Genes ◽

High Rate ◽

Resistance Pattern ◽

Antibiotics Resistance ◽

Disk Diffusion Test ◽

E Coli ◽

Genes Encoding ◽

Tract Infections

Background and Objectives: Escherichia coli is known to be the pathogen commonly isolated from those infected with uri- nary tract infections (UTIs). The aim of this study was to investigate the presence of E. coli virulence genes and antibiotics’ resistance pattern among clinical isolates in the Northeast of Iran. Relationships between virulence genes and antimicrobial resistances were studied as well. Materials and Methods: Three hundred isolates of E. coli were isolated from patients with UTIs that referred to Ghaem and Imam Reza hospitals (Mashhad, Iran) during August 2016 to February 2017. A multiplex PCR was employed to amplify the genes encoding pyelonephritis associated pili (pap), S-family adhesions (sfa), type1fimbriae (fimH) and aerobactin (aer). Disk diffusion test was performed to test the susceptibility of isolates to β-lactams, aminoglycosides, cephalosporins, quino- lone, fluoroquinolones, carbapenems and trimethoprim-sulfamethoxazole. Results: The PCR results identified the fimH in 78.4%, aer in 70.5%, sfa in 13.6% and the pap in 8.2% of isolates. The rates of antibiotic resistance of the isolates were as follows: 64.7% resistant to cephalosporins, 34% to trimethoprim-sul- famethoxazole, 31% to fluoroquinolones, 15.3% to aminoglycosides, 13.3% to β-lactams, 7.8% to quinolones and 4.4% to carbapenems. Significant relationships existed between pap and aer, pap and sfa, aer and fluoroquinolones also pap and cephalosporins. Conclusion: fimH and aer were found in > 50% of isolates suggesting the importance of both genes in UPEC. The majority of isolates had fimH as adhesion factor for colonization. Determining antibiotic resistance patterns in specific geographical areas is necessary for appropriate treatment of urinary tract infection. The high rate of resistance to cephalosporins is most likely due to incorrect drug administration

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Antibiotic resistance and virulence genes of extraintestinal pathogenic Escherichia coli from tropical estuary, south India

The Journal of Infection in Developing Countries ◽

10.3855/jidc.5627 ◽

2015 ◽

Vol 9 (05) ◽

pp. 496-504 ◽

Cited By ~ 1

Author(s):

Divya Sukumaran ◽

Abdulla A Mohamed Hatha

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Multidrug Resistance ◽

Virulence Factor ◽

Virulence Genes ◽

Antimicrobial Agents ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Virulence Factor Genes

Introduction: Escherichia coli strains can cause a variety of intestinal and extraintestinal diseases. Extraintestinal pathogenic E. coli (ExPEC) strains have the ability to cause severe extraintestinal infections. Multidrug resistance among ExPEC could complicate human infections. Methodology: Escherichia coli strains were isolated during the period of January 2010 to December 2012 from five different stations set at Cochin estuary. Susceptibility testing was determined by the disk-diffusion method using nine different antimicrobial agents. A total of 155 strains of Escherichia coli were screened for the presence of virulence factor genes including papAH, papC, sfa/focDE, iutA,and kpsMT II associated with ExPEC. Results: Among the 155 E. coli isolates, 26 (16.77%), carried two or more virulence genes typical of ExPEC. Furthermore, 19.23% of the ExPEC isolates with multidrug resistance were identified to belong to phylogenetic groups B2 and D. Statistically significant association of iutA gene in ExPEC was found with papC (p < 0.001) and kpsMT II (p < 0.001) genes. ExPEC isolates were mainly resistant to ampicillin (23.07%), tetracycline (19.23%), co-trimoxazole (15.38%), and cefotaxime (15.38%). The adhesion genes papAH and sfa/focDE were positively associated with resistance to gentamicin, chloramphenicol, and cefotaxime (p < 0.05). Conclusions: Co-occurrence of virulence factor genes with antibiotic resistance among ExPEC poses considerable threat to those who use this aquatic system for a living and for recreation.

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Genetic Investigation of Beta-Lactam Associated Antibiotic Resistance Among Escherichia Coli Strains Isolated from Water Sources

The Open Microbiology Journal ◽

10.2174/1874285801711010203 ◽

2017 ◽

Vol 11 (1) ◽

pp. 203-210 ◽

Cited By ~ 6

Author(s):

Reza Ranjbar ◽

Mehrdad Sami

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Surface Water ◽

Water Resources ◽

Water Sources ◽

Resistant Bacteria ◽

Beta Lactam ◽

E Coli ◽

Surface Water Resources ◽

Extended Spectrum Beta Lactamases

Background: Antimicrobial resistance is an important factor threatening human health. It is widely accepted that antibiotic resistant bacteria such as Escherichia coli (E. coli) released from humans and animals into the water sources, can introduce their resistance genes into the natural bacterial community. Objective: The aim of this study was to investigate the prevalence of blaTEM, blaCTX, blaSHV, blaOXA and blaVEB associated-antibiotic resistance among E. coli bacteria isolated from different water resources in Iran. Methods: The study contained all E. coli strains segregated from different surface water sources. The Kirby-Bauer method and combined discs method was determined in this study for testing antimicrobial susceptibility and strains that produced Extended-Spectrum Beta Lactamases (ESBL), respectively. DNA extraction kit was applied for genomic and plasmid DNA derivation. Finally the frequency of resistant genes including blaTEM, blaCTX, blaSHV, blaOXA and blaVEB in ESBL producing isolates were studied by PCR. Results: One hundred E. coli strains were isolated and entered in the study. The highest antibiotic resistance was observed on clindamycin (96%). Moreover, 38.5% isolates were ESBL producers. The frequency of different ESBLs genes were 37%, 27%, 27%, and 25% for blaTEM, blaCTX, blaSHV, and blaOXA, respectively. The blaVEB wasn’t found in any isolates. Conclusion: The study revealed a high prevalence of CTX-M, TEM, SHV and OXA genes among E. coli strains in surface water resources. In conclusion, these results raised a concern regarding the presence and distribution of these threatening factors in surface water sources and its subsequent outcomes.

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