Molecular Detection of Antimicrobial Resistance Genes and Virulence Genes in E. coli Isolated from Sheep and Goat Faecal Samples

Background: E. coli is a Gram negative bacterium commonly found in the intestine of animals and humans. Most E. coli are harmless but some can cause infection. Antimicrobial resistance is a public health problem, hence this study was carried out to estimate the antimicrobial resistance pattern and virulotyping of E. coli isolates.Methods: During the period 2018-19, a total of 100 faecal samples from goats (n=50) and sheep (n=50) were processed for isolation and identification of E.coli. All the E. coli isolates were tested for resistance against 10 different antibiotics. The prevalence of various antibiotic resistance genes viz bla TEM, tetA and tetB and virulence genes Stx1, Stx2 and hlyA in all E. coli isolates was determined by PCR.Result: Out of 100 samples, 40 isolates of E.coli (24 from goats and 16 from sheep) were isolated, identified biochemically and by PCR. Majority of the E.coli isolates were found resistant to penicillin (100%) followed by ampicillin (93%), tetracycline, (68.8%), gentamicin (37.4%), ceftriazone (18.8%), amikacin (16.7%), chloramphenicol (4.2%), co- trimoxazole (4.2%) and ofloxacin (4.2%). Out of all the isolates, 12 isolates were positive for blaTEM, 10 for tetB, 19 for tetA, five for Stx1 and hlyA and none carried Stx2 gene.

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Virulence and antimicrobial resistance genes associated with the in-vivo pathogenicity of avian pathogenic E. coli isolates

GMPC Thesis and Opinions Platform ◽

10.51585/gtop.2021.0005 ◽

2021 ◽

Vol 1 (1) ◽

pp. 17-20

Author(s):

Ahmed Abd El-Mawgoud ◽

Azza El-Sawah ◽

Soad Nasef ◽

Al-Hussien Dahshan ◽

Ahmed Ali

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Broiler Chickens ◽

Virulence Genes ◽

Virulence Gene ◽

Health Concern ◽

Pathogenicity Test ◽

E Coli ◽

Antimicrobial Resistance Genes

In the current study, ten avian pathogenic E. coli (APEC) isolates of the most predominant APEC serogroups isolated from broiler chickens in Egypt were screened for their virulence and antimicrobial resistance genes pattern using PCR. Five selected virulence gene patterns were further investigated for their in-vivo pathogenicity test. Results showed a 100% prevalence of the β-lactams and tetracyclines resistance genes. However, aminoglycoside and quinolone resistance genes were not detected. Also, 80% of the tested isolates harbored mcr-1 gene, colistin resistance gene. In-vivo pathogenic strains consistently harbored the virulence gene pattern of fimH, fimA, papC, iutA, and tsh. Additionally, the tsh gene was consistently detected with lethal APEC isolates in day-old chicks. These results highlighted the high prevalence of antimicrobial and virulence genes in APEC that potentially represent a public health concern. In this study, the virulence genes fimH, fimA, papC, iutA, and tsh were the most common virulence gene patterns associated with pathogenicity in day-old chicks.

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A Virulence and Antimicrobial Resistance DNA Microarray Detects a High Frequency of Virulence Genes in Escherichia coli Isolates from Great Lakes Recreational Waters

Applied and Environmental Microbiology ◽

10.1128/aem.00137-06 ◽

2006 ◽

Vol 72 (6) ◽

pp. 4200-4206 ◽

Cited By ~ 82

Author(s):

Katia Hamelin ◽

Guillaume Bruant ◽

Abdel El-Shaarawi ◽

Stephen Hill ◽

Thomas A. Edge ◽

...

Keyword(s):

Public Health ◽

Escherichia Coli ◽

Antimicrobial Resistance ◽

Great Lakes ◽

Dna Microarray ◽

Resistance Genes ◽

Virulence Genes ◽

Recreational Waters ◽

E Coli ◽

Antimicrobial Resistance Genes

ABSTRACT Escherichia coli is generally described as a commensal species with occasional pathogenic strains. Due to technological limitations, there is currently little information concerning the prevalence of pathogenic E. coli strains in the environment. For the first time, using a DNA microarray capable of detecting all currently described virulence genes and commonly found antimicrobial resistance genes, a survey of environmental E. coli isolates from recreational waters was carried out. A high proportion (29%) of 308 isolates from a beach site in the Great Lakes carried a pathotype set of virulence-related genes, and 14% carried antimicrobial resistance genes, findings consistent with a potential risk for public health. The results also showed that another 8% of the isolates had unusual virulence gene combinations that would be missed by conventional screening. This new application of a DNA microarray to environmental waters will likely have an important impact on public health, epidemiology, and microbial ecology in the future.

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Uropathogenic Escherichia coli in the Urine Samples of Iranian Dogs: Antimicrobial Resistance Pattern and Distribution of Antibiotic Resistance Genes

BioMed Research International ◽

10.1155/2017/4180490 ◽

2017 ◽

Vol 2017 ◽

pp. 1-10 ◽

Cited By ~ 1

Author(s):

Amirhossein Yousefi ◽

Saam Torkan

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Urine Samples ◽

Disk Diffusion ◽

Resistance Pattern ◽

E Coli ◽

Tract Infections

Resistant uropathogenic Escherichia coli is the most common cause of urinary tract infections in dogs. The present research was done to study the prevalence rate and antimicrobial resistance properties of UPEC strains isolated from healthy dogs and those which suffered from UTIs. Four-hundred and fifty urine samples were collected and cultured. E. coli-positive strains were subjected to disk diffusion and PCR methods. Two-hundred out of 450 urine samples (44.4%) were positive for E. coli. Prevalence of E. coli in healthy and infected dogs was 28% and 65%, respectively. Female had the higher prevalence of E. coli (P=0.039). Marked seasonality was also observed (P=0.024). UPEC strains had the highest levels of resistance against gentamicin (95%), ampicillin (85%), amikacin (70%), amoxicillin (65%), and sulfamethoxazole-trimethoprim (65%). We found that 21.50% of UPEC strains had simultaneously resistance against more than 10 antibiotics. Aac(3)-IV (77%), CITM (52.5%), tetA (46.5%), and sul1 (40%) were the most commonly detected antibiotic resistance genes. Findings showed considerable levels of antimicrobial resistance among UPEC strains of Iranian dogs. Rapid identification of infected dogs and their treatment based on the results of disk diffusion can control the risk of UPEC strains.

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Associations between Antimicrobial Resistance Phenotypes, Antimicrobial Resistance Genes, and Virulence Genes of Fecal Escherichia coli Isolates from Healthy Grow-Finish Pigs

Applied and Environmental Microbiology ◽

10.1128/aem.01253-08 ◽

2009 ◽

Vol 75 (5) ◽

pp. 1373-1380 ◽

Cited By ~ 51

Author(s):

Leigh B. Rosengren ◽

Cheryl L. Waldner ◽

Richard J. Reid-Smith

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Large Scale ◽

Virulence Genes ◽

Antimicrobial Use ◽

Phenotypic Resistance ◽

E Coli ◽

Antimicrobial Resistance Genes ◽

Antimicrobial Resistance Gene

ABSTRACT Escherichia coli often carries linked antimicrobial resistance genes on transmissible genetic elements. Through coselection, antimicrobial use may select for unrelated but linked resistance or virulence genes. This study used unconditional statistical associations to investigate the relationships between antimicrobial resistance phenotypes and antimicrobial resistance genes in 151 E. coli isolates from healthy pigs. Phenotypic resistance to each drug was significantly associated with phenotypic resistance to at least one other drug, and every association found that the probability of observing the outcome resistance was increased by the presence of the predictor resistance. With one exception, each statistical association that was identified between a pair of resistance genes had a corresponding significant association identified between the phenotypes mediated by those genes. This suggests that associations between resistance phenotypes might predict coselection. If this hypothesis is confirmed, evaluation of the associations between resistance phenotypes could improve our knowledge of coselection dynamics and provide a cost-effective way to evaluate existing data until large-scale genotypic data collection becomes feasible. This could enable policy makers and users of antimicrobials to consider coselection in antimicrobial use decisions. This study also considered the unconditional relationships between resistance and virulence genes in E. coli from healthy pigs (aidA-1, eae, elt, estA, estB, fedA1, stx1, and stx2). Positive statistical associations would suggest that antimicrobial use may select for virulence in bacteria that may contaminate food or cause diarrhea in pigs. Fortunately, the odds of detecting a virulence gene were rarely increased by the presence of an antimicrobial resistance gene. This suggests that on-farm antimicrobial use did not select for the examined virulence factors in E. coli carried by this population of healthy pigs.

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Genes Encoding the Virulence and the Antimicrobial Resistance in Enterotoxigenic and Shiga-toxigenic E. coli Isolated from Diarrheic Calves

Toxins ◽

10.3390/toxins12060383 ◽

2020 ◽

Vol 12 (6) ◽

pp. 383 ◽

Cited By ~ 6

Author(s):

Abdelazeem M. Algammal ◽

Ali El-Kholy ◽

Emad M. Riad ◽

Hossam E. Mohamed ◽

Mahmoud M. Elhaig ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Virulence Genes ◽

Antimicrobial Agents ◽

Economic Losses ◽

Shiga Toxins ◽

E Coli ◽

Calf Diarrhea ◽

Antimicrobial Resistance Genes ◽

Diarrheic Calves

Calf diarrhea is one of the considerable infectious diseases in calves, which results in tremendous economic losses globally. To determine the prevalence of Shiga-toxigenic E. coli (STEC) and Enterotoxigenic E. coli (ETEC) incriminated in calf diarrhea, with special reference to Shiga- toxins genes (stx1 and stx2) and enterotoxins genes (lt and sta) that govern their pathogenesis, as well as the virulence genes; eaeA (intimin) and f41(fimbrial adhesion), and the screening of their antibiogram and antimicrobial resistance genes; aadB, sul1, and bla-TEM, a total of 274 fecal samples were collected (April 2018–Feb 2019) from diarrheic calves at different farms in El-Sharqia Governorate, Egypt. The bacteriological examination revealed that the prevalence of E. coli in diarrheic calves was 28.8%. The serotyping of the isolated E. coli revealed 7 serogroups; O26, O128, O111, O125, O45, O119 and O91. Furthermore, the Congo red binding test was carried out, where 89.8% of the examined strains (n = 71) were positive. The antibiogram of the isolated strains was investigated; the majority of E. coli serotypes exhibit multidrug resistance (MDR) to four antimicrobial agents; neomycin, gentamycin, streptomycin, and amikacin. Polymerase chain reaction (PCR) was used to detect the prevalence of the virulence genes; stx1, stx2 lt, sta, f41 and eaeA, as well as the antimicrobial resistance genes; aadB, sul1, and bla-TEM. The prevalence of STEC was 20.2% (n = 16), while the prevalence of ETEC was 30.4% (n = 24). Briefly, the Shiga toxins genes; stx1 and stx2, are the most prevalent virulence genes associated with STEC, which are responsible for the pathogenesis of the disease and helped by the intimin gene (eaeA). In addition, the lt gene is the most prevalent enterotoxin gene accompanied by the ETEC strains, either alone or in combination with sta and/or f41 genes. The majority of pathogenic E. coli incriminated in calf diarrhea possesses the aadB resistance gene, followed by the sul1 gene. Enrofloxacin, florfenicol, amoxicillin-clavulanic acid, and ampicillin-sulbactam, are the most effective antimicrobial agents against the isolated STEC and ETEC strains.

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Virulence Properties of mcr-1-Positive Escherichia coli Isolated from Retail Poultry Meat

Microorganisms ◽

10.3390/microorganisms9020308 ◽

2021 ◽

Vol 9 (2) ◽

pp. 308

Author(s):

Michaela Kubelová ◽

Ivana Koláčková ◽

Tereza Gelbíčová ◽

Martina Florianová ◽

Alžběta Kalová ◽

...

Keyword(s):

Escherichia Coli ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Poultry Meat ◽

Human Populations ◽

E Coli ◽

Antimicrobial Resistance Genes ◽

Mlst Type ◽

On Line ◽

Sequence Types

The great plasticity and diversity of the Escherichia coli genome, together with the ubiquitous occurrence, make E. coli a bacterium of world-wide concern. Of particular interest are pathogenic strains and strains harboring antimicrobial resistance genes. Overlapping virulence-associated traits between avian-source E. coli and human extraintestinal pathogenic E. coli (ExPEC) suggest zoonotic potential and safety threat of poultry food products. We analyzed whole-genome sequencing (WGS) data of 46 mcr-1-positive E. coli strains isolated from retail raw meat purchased in the Czech Republic. The investigated strains were characterized by their phylogroup—B1 (43%), A (30%), D (11%), E (7%), F (4%), B2 (2%), C (2%), MLST type, and serotype. A total of 30 multilocus sequence types (STs), of which ST744 was the most common (11%), were identified, with O8 and O89 as the most prevalent serogroups. Using the VirulenceFinder tool, 3 to 26 virulence genes were detected in the examined strains and a total of 7 (15%) strains met the pathogenic criteria for ExPEC. Four strains were defined as UPEC (9%) and 18 (39%) E. coli strains could be classified as APEC. The WGS methods and available on-line tools for their evaluation enable a comprehensive approach to the diagnosis of virulent properties of E. coli strains and represent a suitable and comfortable platform for their detection. Our results show that poultry meat may serve as an important reservoir of strains carrying both virulence and antibiotic resistance genes for animal and human populations.

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Serotype distribution, antimicrobial susceptibility, antimicrobial resistance genes and virulence genes of Salmonella isolated from a pig slaughterhouse in Yangzhou, China

AMB Express ◽

10.1186/s13568-019-0936-9 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Quan Li ◽

Jian Yin ◽

Zheng Li ◽

Zewei Li ◽

Yuanzhao Du ◽

...

Keyword(s):

Public Health ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Antimicrobial Susceptibility ◽

Virulence Genes ◽

Multidrug Resistant ◽

Resistance Rate ◽

Economic Losses ◽

Production Chain ◽

Antimicrobial Resistance Genes

AbstractSalmonella is an important food-borne pathogen associated with public health and high economic losses. To investigate the prevalence and the characteristics of Salmonella in a pig slaughterhouse in Yangzhou, a total of 80 Salmonella isolates were isolated from 459 (17.43%) samples in 2016–2017. S. Derby (35/80, 43.75%) was the most prevalent, followed by S. Rissen (16/80, 20.00%) and S. Newlands (11/80, 13.75%). The highest rates of susceptibility were observed to cefoxitin (80/80, 100.0%) and amikacin (80/80, 100.0%), followed by aztreonam (79/80, 98.75%) and nitrofurantoin (79/80, 98.75%). The highest resistance rate was detected for tetracycline (65/80, 81.25%), followed by ampicillin (60/80, 75.00%), bactrim (55/80, 68.75%), and sulfisoxazole (54/80, 67.50%). Overall, 91.25% (73/80) of the isolates were resistant to at least one antibiotic, while 71.25% (57/80) of the isolate strains were multidrug resistant in the antimicrobial susceptibility tested. In addition, 86.36% (19/22) of the 22 antimicrobial resistance genes in the isolates were identified. Our data indicated that the resistance to certain antimicrobials was significantly associated, in part, with antimicrobial resistance genes. Furthermore, 81.25% (65/80) isolates harbored the virulence gene of mogA, of which 2 Salmonella Typhimurium isolates carried the mogA, spvB and spvC virulence genes at the same time. The results showed that swine products in the slaughterhouse were contaminated with multidrug resistant Salmonella commonly, especially some isolates carry the spv virulence genes. The virulence genes might facilitate the dissemination of the resistance genes to consumers along the production chain, suggesting the importance of controlling Salmonella during slaughter for public health.

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Multi-omic surveillance of Escherichia coli and Klebsiella spp. in hospital sink drains and patients

10.1101/2020.02.19.952366 ◽

2020 ◽

Author(s):

B Constantinides ◽

KK Chau ◽

TP Quan ◽

G Rodger ◽

M Andersson ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Bacterial Infections ◽

Wide Spectrum ◽

Klebsiella Oxytoca ◽

Clinical Disease ◽

E Coli ◽

Antimicrobial Resistance Genes ◽

Wide Range

ABSTRACTEscherichia coli and Klebsiella spp. are important human pathogens that cause a wide spectrum of clinical disease. In healthcare settings, sinks and other wastewater sites have been shown to be reservoirs of antimicrobial-resistant E. coli and Klebsiella spp., particularly in the context of outbreaks of resistant strains amongst patients. Without focusing exclusively on resistance markers or a clinical outbreak, we demonstrate that many hospital sink drains are abundantly and persistently colonised with diverse populations of E. coli, Klebsiella pneumoniae and Klebsiella oxytoca, including both antimicrobial-resistant and susceptible strains. Using whole genome sequencing (WGS) of 439 isolates, we show that environmental bacterial populations are largely structured by ward and sink, with only a handful of lineages, such as E. coli ST635, being widely distributed, suggesting different prevailing ecologies which may vary as a result of different inputs and selection pressures. WGS of 46 contemporaneous patient isolates identified one (2%; 95% CI 0.05-11%) E. coli urine infection-associated isolate with high similarity to a prior sink isolate, suggesting that sinks may contribute to up to 10% of infections caused by these organisms in patients on the ward over the same timeframe. Using metagenomics from 20 sink-timepoints, we show that sinks also harbour many clinically relevant antimicrobial resistance genes including blaCTX-M, blaSHV and mcr, and may act as niches for the exchange and amplification of these genes. Our study reinforces the potential role of sinks in contributing to Enterobacterales infection and antimicrobial resistance in hospital patients, something that could be amenable to intervention.IMPORTANCEEscherichia coli and Klebsiella spp. cause a wide range of bacterial infections, including bloodstream, urine and lung infections. Previous studies have shown that sink drains in hospitals may be part of transmission chains in outbreaks of antimicrobial-resistant E. coli and Klebsiella spp., leading to colonisation and clinical disease in patients. We show that even in non-outbreak settings, contamination of sink drains by these bacteria is common across hospital wards, and that many antimicrobial resistance genes can be found and potentially exchanged in these sink drain sites. Our findings demonstrate that the colonisation of handwashing sink drains by these bacteria in hospitals is likely contributing to some infections in patients, and that additional work is needed to further quantify this risk, and to consider appropriate mitigating interventions.

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An Evaluation of the Pathogenic Potential, and the Antimicrobial Resistance, of Salmonella Strains Isolated from Mussels

Microorganisms ◽

10.3390/microorganisms10010126 ◽

2022 ◽

Vol 10 (1) ◽

pp. 126

Author(s):

Antonio Lozano-León ◽

Carlos García-Omil ◽

Rafael R. Rodríguez-Souto ◽

Alexandre Lamas ◽

Alejandro Garrido-Maestu

Keyword(s):

Antimicrobial Resistance ◽

Resistance Genes ◽

Virulence Genes ◽

Health Issues ◽

Salmonella Spp ◽

Pathogenic Potential ◽

Antimicrobial Resistance Genes ◽

Important Health ◽

Main Production ◽

Production Areas

Salmonella spp. and antimicrobial resistant microorganisms are two of the most important health issues worldwide. In the present study, strains naturally isolated from mussels harvested in Galicia (one of the main production areas in the world), were genetically characterized attending to the presence of virulence and antimicrobial resistance genes. Additionally, the antimicrobial profile was also determined phenotypically. Strains presenting several virulence genes were isolated but lacked all the antimicrobial resistance genes analyzed. The fact that some of these strains presented multidrug resistance, highlighted the possibility of bearing different genes than those analyzed, or resistance based on completely different mechanisms. The current study highlights the importance of constant surveillance in order to improve the safety of foods.

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Molecular characterisation of antimicrobial resistance and virulence genes in Escherichia coli strains isolated from diarrhoeic and healthy rabbits in Tunisia

World Rabbit Science ◽

10.4995/wrs.2020.10879 ◽

2020 ◽

Vol 28 (2) ◽

pp. 81

Author(s):

Raouia Ben Rhouma ◽

Ahlem Jouini ◽

Amira Klibi ◽

Safa Hamrouni ◽

Aziza Boubaker ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Virulence Genes ◽

Antimicrobial Agents ◽

Antibiotic Resistance Genes ◽

Virulence Gene ◽

Diffusion Method ◽

Class 1 Integron ◽

E Coli ◽

Class 1

The purpose of this study was to identify Escherichia coli isolates in diarrhoeic and healthy rabbits in Tunisia and characterise their virulence and antibiotic resistance genes. In the 2014-2015 period, 60 faecal samples from diarrhoeic and healthy rabbits were collected from different breeding farms in Tunisia. Susceptibility to 14 antimicrobial agents was tested by disc diffusion method and the mechanisms of gene resistance were evaluated using polymerase chain reaction and sequencing methods. Forty E. coli isolates were recovered in selective media. High frequency of resistance to tetracycline (95%) was detected, followed by different levels of resistance to sulphonamide (72.5%), streptomycin (62.5%), trimethoprim-sulfamethoxazole (60%), nalidixic acid (32.5%), ampicillin (37.5%) and ticarcillin (35%). E. coli strains were susceptible to cefotaxime, ceftazidime and imipenem. Different variants of blaTEM, tet, sul genes were detected in most of the strains resistant to ampicillin, tetracycline and sulphonamide, respectively. The presence of class 1 integron was studied in 29 sulphonamide-resistant E. coli strains from which 15 harboured class 1 integron with four different arrangements of gene cassettes, dfrA17+aadA5 (n=9), dfrA1 + aadA1 (n=4), dfrA12 + addA2 (n=1), dfrA12+orf+addA2 (n=1). The qnrB gene was detected in six strains out of 13 quinolone-resistant E. coli strains. Seventeen E. coli isolates from diarrhoeic rabbits harboured the enteropathogenic eae genes associated with different virulence genes tested (fimA, cnf1, aer), and affiliated to B2 (n=8) and D (n=9) phylogroups. Isolated E. coli strains from healthy rabbit were harbouring fim A and/or cnf1 genes and affiliated to A and B1 phylogroups. This study showed that E. coli strains from the intestinal tract of rabbits are resistant to the widely prescribed antibiotics in medicine. Therefore, they constitute a reservoir of antimicrobial-resistant genes, which may play a significant role in the spread of antimicrobial resistance. In addition, the eae virulence gene seemed to be implicated in diarrhoea in breeder rabbits in Tunisia.

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