scholarly journals Characterisation of Early Positive mcr-1 Resistance Gene and Plasmidome in Escherichia coli Pathogenic Strains Associated with Variable Phylogroups under Colistin Selection

Antibiotics ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1041
Author(s):  
Guerrino Macori ◽  
Scott V. Nguyen ◽  
Ankita Naithani ◽  
Daniel Hurley ◽  
Li Bai ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Single Molecule ◽  
Bovine Mastitis ◽  
Antibiotic Resistance Genes ◽  
Genomic Context ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Faecal Samples ◽  
Long Read

An antibiotic susceptibility monitoring programme was conducted from 2004 to 2010, resulting in a collection of 143 Escherichia coli cultured from bovine faecal samples (diarrhoea) and milk-aliquots (mastitis). The isolates were subjected to whole-genome sequencing and were distributed in phylogroups A, B1, B2, C, D, E, and G with no correlation for particular genotypes with pathotypes. In fact, the population structure showed that the strains belonging to the different phylogroups matched broadly to ST complexes; however, the isolates are randomly associated with the diseases, highlighting the necessity to investigate the virulence factors more accurately in order to identify the mechanisms by which they cause disease. The antimicrobial resistance was assessed phenotypically, confirming the genomic prediction on three isolates that were resistant to colistin, although one isolate was positive for the presence of the gene mcr-1 but susceptible to colistin. To further characterise the genomic context, the four strains were sequenced by using a single-molecule long read approach. Genetic analyses indicated that these four isolates harboured complex and diverse plasmids encoding not only antibiotic resistant genes (including mcr-1 and bla) but also virulence genes (siderophore, ColV, T4SS). A detailed description of the plasmids of these four E. coli strains, which are linked to bovine mastitis and diarrhoea, is presented for the first time along with the characterisation of the predicted antibiotic resistance genes. The study highlighted the diversity of incompatibility types encoding complex antibiotic resistance elements such as Tn6330, ISEcp1, Tn6029, and IS5075. The mcr-1 resistance determinant was identified in IncHI2 plasmids pCFS3273-1 and pCFS3292-1, thus providing some of the earliest examples of mcr-1 reported in Europe, and these sequences may be a representative of the early mcr-1 plasmidome characterisation in the EU/EEA.

scholarly journals Mechanisms of Resistance in Multiple-Antibiotic-Resistant Escherichia coli Strains of Human, Animal, and Food Origins

2004 ◽  
Vol 48 (10) ◽  
pp. 3996-4001 ◽  
Author(s):  
Yolanda Sáenz ◽  
Laura Briñas ◽  
Elena Domínguez ◽  
Joaquim Ruiz ◽  
Myriam Zarazaga ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Amino Acid ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Mechanisms Of Resistance ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Class 1 ◽  
Human Animal

ABSTRACT Seventeen multiple-antibiotic-resistant nonpathogenic Escherichia coli strains of human, animal, and food origins showed a wide variety of antibiotic resistance genes, many of them carried by class 1 and class 2 integrons. Amino acid changes in MarR and mutations in marO were identified for 15 and 14 E. coli strains, respectively.

scholarly journals Antibiotic-resistant Escherichia coli and Salmonella spp. associated with dairy cattle and farm environment having public health significance

2019 ◽  
Vol 12 (7) ◽  
pp. 984-993 ◽  
Author(s):  
Md. Abdus Sobur ◽  
Abdullah Al Momen Sabuj ◽  
Ripon Sarker ◽  
A. M. M. Taufiqur Rahman ◽  
S. M. Lutful Kabir ◽  
...  
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
One Health ◽  
Dairy Farm ◽  
Antibiotic Resistance Genes ◽  
Salmonella Spp ◽  
Antibiotic Resistant ◽  
E Coli

Aim: The present study was carried out to determine load of total bacteria, Escherichia coli and Salmonella spp. in dairy farm and its environmental components. In addition, the antibiogram profile of the isolated bacteria having public health impact was also determined along with identification of virulence and resistance genes by polymerase chain reaction (PCR) under a one-health approach. Materials and Methods: A total of 240 samples of six types (cow dung - 15, milk - 10, milkers' hand wash - 10, soil - 10 water - 5, and vegetables - 10) were collected from four dairy farms. For enumeration, the samples were cultured onto plate count agar, eosin methylene blue, and xylose-lysine deoxycholate agar and the isolation and identification of the E. coli and Salmonella spp. were performed based on morphology, cultural, staining, and biochemical properties followed by PCR. The pathogenic strains of E. coli stx1, stx2, and rfbO157 were also identified through PCR. The isolates were subjected to antimicrobial susceptibility test against 12 commonly used antibiotics by disk diffusion method. Detection of antibiotic resistance genes ereA, tetA, tetB, and SHV were performed by PCR. Results: The mean total bacterial count, E. coli and Salmonella spp. count in the samples ranged from 4.54±0.05 to 8.65±0.06, 3.62±0.07 to 7.04±0.48, and 2.52±0.08 to 5.87±0.05 log colony-forming unit/g or ml, respectively. Out of 240 samples, 180 (75%) isolates of E. coli and 136 (56.67%) isolates of Salmonella spp. were recovered through cultural and molecular tests. Among the 180 E. coli isolates, 47 (26.11%) were found positive for the presence of all the three virulent genes, of which stx1 was the most prevalent (13.33%). Only three isolates were identified as enterohemorrhagic E. coli. Antibiotic sensitivity test revealed that both E. coli and Salmonella spp. were found highly resistant to azithromycin, tetracycline, erythromycin, oxytetracycline, and ertapenem and susceptible to gentamycin, ciprofloxacin, and imipenem. Among the four antibiotic resistance genes, the most observable was tetA (80.51-84.74%) in E. coli and Salmonella spp. and SHV genes were the lowest one (22.06-25%). Conclusion: Dairy farm and their environmental components carry antibiotic-resistant pathogenic E. coli and Salmonella spp. that are potential threat for human health which requires a one-health approach to combat the threat.

scholarly journals Survey and Sequence Characterization of Bovine Mastitis-Associated Escherichia coli in Dairy Herds

2020 ◽  
Vol 7 ◽  
Author(s):  
John I. Alawneh ◽  
Ben Vezina ◽  
Hena R. Ramay ◽  
Hulayyil Al-Harbi ◽  
Ameh S. James ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Bovine Mastitis ◽  
Antibiotic Resistance Genes ◽  
Tetracycline Resistance ◽  
Sequence Type ◽  
Genotypic Differences ◽  
Beta Lactam ◽  
Sequence Characterization ◽  
E Coli

Escherichia coli is frequently associated with mastitis in cattle. “Pathogenic” and “commensal” isolates appear to be genetically similar. With a few exceptions, no notable genotypic differences have been found between commensal and mastitis-associated E. coli. In this study, 24 E. coli strains were isolated from dairy cows with clinical mastitis in three geographic regions of Australia (North Queensland, South Queensland, and Victoria), sequenced, then genomically surveyed. There was no observed relationship between sequence type (ST) and region (p = 0.51). The most common Multi Locus Sequence Type was ST10 (38%), then ST4429 (13%). Pangenomic analysis revealed a soft-core genome of 3,463 genes, including genes associated with antibiotic resistance, chemotaxis, motility, adhesion, biofilm formation, and pili. A total of 36 different plasmids were identified and generally found to have local distributions (p = 0.02). Only 2 plasmids contained antibiotic resistance genes, a p1303_5-like plasmid encoding multidrug-resistance (trimethoprim, quaternary ammonium, beta-lactam, streptomycin, sulfonamide, and kanamycin) from two North Queensland isolates on the same farm, while three Victorian isolates from the same farm contained a pCFSAN004177P_01-like plasmid encoding tetracycline-resistance. This pattern is consistent with a local spread of antibiotic resistance through plasmids of bovine mastitis cases. Notably, co-occurrence of plasmids containing virulence factors/antibiotic resistance with putative mobilization was rare, though the multidrug resistant p1303_5-like plasmid was predicted to be conjugative and is of some concern. This survey has provided greater understanding of antibiotic resistance within E. coli-associated bovine mastitis which will allow greater prediction and improved decision making in disease management.

Characterization of Antibiotic Resistance E. Coli and Antibiotic Resistance Genes in Aquatic Environment of Taihu Lake, China

2013 ◽  
Vol 295-298 ◽  
pp. 630-634 ◽  
Author(s):  
Ni Ni Han ◽  
Song He Zhang ◽  
Pei Fang Wang ◽  
Chao Wang
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Surface Water ◽  
Resistance Genes ◽  
Taihu Lake ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Realtime Pcr

The aims of this study are to evaluate multiple antibiotic resistant Escherichia coli isolated from surface water and to investigate the presence and distribution antibiotic resistance genes (ARGs) in sediments of Taihu Lake. The results show that the presentence of four ARGs concentrations in the sediments of the lake was in sequence: strB>qnrB>strA>qnrS, as determined by realtime-PCR technique. The southwest and east areas of Taihu Lake were polluted seriously than other areas from all kinds of antibiotics. The screening Escherichia coli had a higher resistance to streptomycin, tetracycline and ampicillin than other four antibiotics, and had a lowest resistance to levofloxacin.

scholarly journals Detection of Antibiotic Resistance Genes in Some Multiple Antibiotic Resistant E. coli from Apparently Healthy Pregnant Women

2020 ◽  
pp. 23-31
Author(s):  
O. C. Adekunle ◽  
A. J. Falade- Fatila ◽  
R. Ojedele ◽  
G. Odewale
Keyword(s):  
Escherichia Coli ◽  
Drug Resistance ◽  
Antibiotic Resistance ◽  
Pregnant Women ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Multi Drug Resistance ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Apparently Healthy

The emerging drug resistance, especially among the Escherichia coli (E.coli) isolates from pregnant women, spread rapidly within the community. Urinary tract infection (UTI) is a well-known bacterial infection posing serious health problem in pregnant women. Also, multi-drug resistance is becoming rampant, and it is of serious public health concern. Treatment of E. coli is now a challenge due to continuous increase in resistance towards commonly prescribed antibiotics, thus posing a threat to treatment. Hence, the aim of the study is to determine antibiotic resistance genes in some multiple antibiotic resistant E.coli from apparently healthy pregnant women in Osun State. A cross-sectional study design was used to collect 150 mid-stream urine samples from apparently healthy pregnant women from March, 2018 to September, 2018. A well structured questionnaire and informed consent were used for data collection. Standard loop technique was used to place 0.001 ml of urine on Cysteine Lactose Electrolyte Deficient (CLED) medium, Blood agar, MacConkey agar and incubated at 37 °C for 24 h. A standard agar disc diffusion method was used to determine antimicrobial susceptibility pattern of the isolates. The molecular detection of the resistant genes was done using PCR techniques. The ages of women enrolled in this study ranges from 22 to 42 years (mean ± standard deviation = 31 ± 4.7 years). Escherichia coli showed high percentage of resistance to ampicillin and low resistance to ciprofloxacin and penicillin. All the E. coli isolates were sensitive to levofloxacin, and most were resistant to Meropenem. Multiple drug resistance was observed in all the isolates. Resistance genes in VIM 390bp, bla ctx-M 585bp and TEM 517bp were detected in some of the representative E. coli isolates profiled. This study identified the presence of Multi-drug resistance genes in E. coli associated UTI among pregnant women in Osogbo.

scholarly journals Recovering Escherichia coli plasmids in the absence of long-read sequencing data

2021 ◽  
Author(s):  
Julian A. Paganini ◽  
Nienke L. Plantinga ◽  
Sergio Arredondo-Alonso ◽  
Rob J.L. Willems ◽  
Anita C. Schurch
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Special Focus ◽  
Valuable Insight ◽  
Sequencing Data ◽  
Data Set ◽  
E Coli ◽  
Long Read

The incidence of infections caused by multidrug-resistant Escherichia coli strains has risen in the past years. Antibiotic resistance in E. coli is often mediated by acquisition and maintenance of plasmids. The study of E. coli plasmid epidemiology and genomics often requires long-read sequencing information, but recently a number of tools that allow plasmid prediction from short-read data have been developed. Here, we reviewed 25 available plasmid prediction tools and categorized them into binary plasmid/chromosome classification tools and plasmid reconstruction tools. We benchmarked six tools that aim to reliably reconstruct distinct plasmids, with a special focus on plasmids carrying antibiotic resistance genes (ARGs) such as extended-spectrum beta-lactamase genes. They use either assembly graph information (plasmidSPAdes, gplas), reference databases (MOB-Suite, FishingForPlasmids) or both (HyAsP and SCAPP) to produce plasmid predictions. The benchmark data set consisted of 240 E. coli strains, harboring 631 plasmids, which were representative for the diversity of E. coli in public databases. Notably, these strains were not used for training any of the tools. We found that two thirds (n=425, 66.3.%) of all plasmids were correctly reconstructed by at least one of the six tools, with a range of 92 (14.58%) to 317 (50.23%) correctly predicted plasmids. However, the majority of plasmids that carried antibiotic resistance genes (n=85, 57.8%) could not be completely recovered as distinct plasmids by any of the tools. MOB-suite was the only tool that was able to correctly reconstruct the majority of plasmids (n=317, 50.23%), and performed best at reconstructing large plasmids (n=166, 46.37%) and ARG-plasmids (n=41, 27.9%), but predictions frequently contained chromosome contamination (40%). In contrast, plasmidSPAdes reconstructed the highest fraction of plasmids smaller than 18 kbp (n=168, 61.54%). Large ARG-plasmids, however, were recovered with small precision values (median=0.47, IQR=0.61), indicating that plasmidSPAdes frequently merged sequences derived from distinct replicons. Additionally, only 63% of all plasmid-borne ARGs were correctly predicted by plasmidSPAdes. The remaining four tools (FishingForPlasmids, HyAsP, SCAPP and gplas) were able to correctly reconstruct a combined total of 18 plasmids that were missed by MOB-suite and plasmidSPAdes. Available bioinformatic tools can provide valuable insight into E. coli plasmids, but also have important limitations. This work will serve as a guideline for selecting the most appropriate plasmid reconstruction tool for studies focusing on E. coli plasmids in the absence of long-read sequencing data.

scholarly journals Isolation and molecular characterization of multidrug-resistant Escherichia coli from chicken meat

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Md. Masudur Rahman ◽  
Asmaul Husna ◽  
Hatem A. Elshabrawy ◽  
Jahangir Alam ◽  
Nurjahan Yasmin Runa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Broiler Chicken ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Chicken Meat ◽  
The Other ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Layer Chicken

AbstractAntibiotic-resistant Escherichia coli (E. coli) are common in retail poultry products. In this study, we aimed to isolate and characterize multidrug resistant (MDR) E. coli in raw chicken meat samples collected from poultry shops in Sylhet division, Bangladesh, as well as to determine correlation between resistance phenotype and genotype. A total of 600 chicken meat swabs (divided equally between broiler and layer farms, n = 300 each) were collected and the isolates identified as E. coli (n = 381) were selected. Disc diffusion antimicrobial susceptibility assay showed resistance of these isolates to ampicillin, erythromycin, tetracycline, streptomycin, trimethoprim-sulfamethoxazole, chloramphenicol, and gentamicin. Polymerase chain reaction (PCR) identified several antibiotic resistance genes (ARGs) in our isolates. Among these ARGs, the prevalence of tetA (for tetracycline) was the highest (72.58%) in broiler chicken isolates, followed by sul1 (for sulfonamide; 44.16%), aadA1 (for streptomycin; 33.50%), ereA (for erythromycin; 27.41%), aac-3-IV (for gentamicin; 25.38%), and the two genes cmlA (24.87%) and catA1 (8.63%) for chloramphenicol. On the other hand, the respective prevalence in layer chicken isolates were 82.06%, 47.83%, 35.87%, 35.33%, 23.91%, 19.02%, and 5.43%. Furthermore, 49.23% of the isolates from broiler chicken were MDR, with the presence of multiple antibiotic resistance genes, including 3 (40.11%) and 4 (9.13%) genes. On the other hand, 51.09% of layer chicken E. coli isolates were MDR, with 3, 4 or 5 ARGs detected in 36.41%, 14.13%, and 0.54% of the isolates, respectively. We also found that 12.8% of broiler chicken E. coli isolates and 7.61% of layer chicken isolates carried genes coding for extended-spectrum SHV beta-lactamases. Lastly, we report the presence of the AmpC beta-lactamase producing gene (CITM) in 4.56% and 3.26% of broiler and layer chicken E. coli isolates, respectively. We found significant correlations between most of the antimicrobial resistant phenotypes and genotypes observed among the investigated E. coli isolates. Our findings highlight the need for the prudent use of antimicrobials in chickens to minimize the development of antibiotic-resistant bacterial strains.

scholarly journals Effect of Urban Wastewater Discharge on the Abundance of Antibiotic Resistance Genes and Antibiotic-Resistant Escherichia coli in Two Italian Rivers

2020 ◽  
Vol 17 (18) ◽  
pp. 6813 ◽  
Author(s):  
Fabrizio Pantanella ◽  
Itziar Lekunberri ◽  
Antonella Gagliardi ◽  
Giuseppe Venuto ◽  
Alexandre Sànchez-Melsió ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Treated Wastewater ◽  
Flow Volume ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Absolute Concentrations ◽  
The Impact

Background: Wastewater treatment plants (WWTPs) are microbial factories aimed to reduce the amount of nutrients and pathogenic microorganisms in the treated wastewater before its discharge into the environment. We studied the impact of urban WWTP effluents on the abundance of antibiotic resistance genes (ARGs) and antibiotic-resistant Escherichia coli (AR-E. coli) in the last stretch of two rivers (Arrone and Tiber) in Central Italy that differ in size and flow volume. Methods: Water samples were collected in three seasons upstream and downstream of the WWTP, at the WWTP outlet, and at sea sites near the river mouth, and analyzed for the abundance of ARGs by qPCR and AR-E. coli using cultivation followed by disk diffusion assays. Results: For all studied genes (16S rRNA, intI1, sul1, ermB, blaTEM, tetW and qnrS), absolute concentrations were significantly higher in the Tiber than in the Arrone at all sampling sites, despite their collection date, but the prevalence of target ARGs within bacterial communities in both rivers was similar. The absolute concentrations of most ARGs were also generally higher in the WWTP effluent with median levels between log 4 and log 6 copies per ml but did not show differences along the studied stretches of rivers. Statistically significant site effect was found for E. coli phenotypic resistance to tetracycline and ciprofloxacin in the Arrone but not in the Tiber. Conclusions: In both rivers, diffuse or point pollution sources other than the studied WWTP effluents may account for the observed resistance pattern, although the Arrone appears as more sensitive to the wastewater impact considering its lower flow volume.

scholarly journals Predominance of blaTEM and tetA genes in antibiotic-resistant Escherichia coli isolates from Laguna Lake, Philippines

2021 ◽  
Author(s):  
Daile Meek Salvador-Membreve ◽  
Windell L. Rivera
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
The Philippines ◽  
Resistant Bacteria ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Laguna Lake ◽  
Antibiotic Genes

Abstract Lakes are one of the sinks of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs); however, information on ARB and ARGs in lakes in the Philippines is scarce. In this study, Escherichia coli was isolated from the largest freshwater lake in the Philippines, Laguna Lake, to detect antibiotic resistance and the presence of ARGs. Broth microdilution assay (BMA) and molecular identification of five environmentally prevalent ARGs (strA, blaCTX-M, blaSHV, blaTEM, and tetA) were performed. The majority (75.70%) of the isolates harbored at least one of the targeted antibiotic genes. Multiplex PCR detected about 49.07% of the isolates had genes for extended-spectrum β-lactamases (ESBL), which were mostly represented by blaTEM (47.66%). The genes strA and tetA were observed in this study with detection frequencies of 29.91 and 45.33%, respectively. About 95.69% of thermotolerant E. coli isolates were non-susceptible to six different antibiotics using BMA. Nearly 37% of the isolates were found to be multidrug-resistant (MDR) with most isolates resistant to ampicillin (81.72%). Furthermore, the occurrence of ESBL genes was significantly correlated with tetA genes (P = 0.013). To date, this study is the first to report on the presence of MDR and thermotolerant E. coli in Laguna Lake, Philippines.

scholarly journals Phenotypic and genotypic characterization of antibiotic-resistant in Escherichia coli isolates from patients with diarrhea

2019 ◽  
Author(s):  
Mojtaba Bonyadian ◽  
Sara Barati ◽  
Mohammad Reza Mahzounieh
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Diffusion Method ◽  
Main Concern ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Resis Tance ◽  
Stool Samples

Background and Objectives: Escherichia coli is a common enteric pathogen of human and livevestock. Antibiotic resis- tance is the main concern of public health. The aim of this study was to detect this bacterium in stool samples of diarrheal patients and identify the phenotypic and genotypic characterizations of antibiotic-resistant isolates such as dfrA1, sul1, citm, tetA, qnr, aac(3)-IV in Shahrekord. Materials and Methods: Two hundred fifty diarrheal stool samples from patients were collected. Microbiological and biochemical examinations were done to detect E. coli. Phenotypic and genotypic antibiotic resistance of the isolates were determined using dick diffusion method and polymerase chain reaction (PCR), respectively. Results: Among 114 E. coli isolates, the least resistance was for gentamicin (0%) and the most resistance was for trimetho- prim (79.8%). The resistance to sulfamethoxazole, ciprofloxacin, ampicillin, and tetracycline were 71.05%, 10.5%, 52.63%, and 3.5% respectively. The results of PCR assay revealed that 10 isolates contain sul1, 49 isolates harbor citm, 8 isolates tetA, 36 isolates dfrA1, 11 isolates qnr genes but there was no isolate with aac(3)-IV gene. In comparison between phenotypic and genotypic of the isolates revealed that citm, tetA, dfrA1, qnr, sul1, aac(3)-IV genes covered 42.98%, 7.01%, 31.57%, 9.64%, 8.7%, 0% of the antibiotic resistance, respectively. Conclusion: Our results revealed that all isolates harbor one or more antibiotic resistance genes and that the PCR is a fast practical and appropriate method to determine the presence of antibiotic resistance genes.

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