scholarly journals Transferable Extended-Spectrum β-Lactamase (ESBL) Plasmids in Enterobacteriaceae from Irrigation Water

2020 ◽  
Vol 8 (7) ◽  
pp. 978
Author(s):  
Maria-Theresia Gekenidis ◽  
Anita Kläui ◽  
Kornelia Smalla ◽  
David Drissner

Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae are classified as serious threats to human health by the U.S. Centers for Disease Control and Prevention. Water used for irrigation of fresh produce can transmit such resistant bacteria directly to edible plant parts. We screened ESBL-producing Escherichia coli, Enterobacter cloacae, and Citrobacter freundii isolated from irrigation water for their potential to transmit resistance to antibiotic-susceptible E. coli. All strains were genome-sequenced and tested in vitro for transmission of resistance to third-generation cephalosporins on solid agar as well as in liquid culture. Of the 19 screened isolates, five ESBL-producing E. coli were able to transfer resistance with different efficiency to susceptible recipient E. coli. Transconjugant strains were sequenced for detection of transferred antibiotic resistance genes (ARGs) and compared to the known ARG pattern of their respective donors. Additionally, phenotypic resistance patterns were obtained for both transconjugant and corresponding donor strains, confirming ESBL-producing phenotypes of all obtained transconjugants.

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kaitlin S. Witherell ◽  
Jason Price ◽  
Ashok D. Bandaranayake ◽  
James Olson ◽  
Douglas R. Call

AbstractMultidrug-resistant bacteria are a growing global concern, and with increasingly prevalent resistance to last line antibiotics such as colistin, it is imperative that alternative treatment options are identified. Herein we investigated the mechanism of action of a novel antimicrobial peptide (CDP-B11) and its effectiveness against multidrug-resistant bacteria including Escherichia coli #0346, which harbors multiple antibiotic-resistance genes, including mobilized colistin resistance gene (mcr-1). Bacterial membrane potential and membrane integrity assays, measured by flow cytometry, were used to test membrane disruption. Bacterial growth inhibition assays and time to kill assays measured the effectiveness of CDP-B11 alone and in combination with colistin against E. coli #0346 and other bacteria. Hemolysis assays were used to quantify the hemolytic effects of CDP-B11 alone and in combination with colistin. Findings show CDP-B11 disrupts the outer membrane of E. coli #0346. CDP-B11 with colistin inhibits the growth of E. coli #0346 at ≥ 10× lower colistin concentrations compared to colistin alone in Mueller–Hinton media and M9 media. Growth is significantly inhibited in other clinically relevant strains, such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae. In rich media and minimal media, the drug combination kills bacteria at a lower colistin concentration (1.25 μg/mL) compared to colistin alone (2.5 μg/mL). In minimal media, the combination is bactericidal with killing accelerated by up to 2 h compared to colistin alone. Importantly, no significant red blood hemolysis is evident for CDP-B11 alone or in combination with colistin. The characteristics of CDP-B11 presented here indicate that it can be used as a potential monotherapy or as combination therapy with colistin for the treatment of multidrug-resistant infections, including colistin-resistant infections.


Pharmacia ◽  
2021 ◽  
Vol 68 (1) ◽  
pp. 1-7
Author(s):  
Hawraa Mohammed Al-Rafyai ◽  
Mourouge Saadi Alwash ◽  
Noor Salman Al-Khafaji

Aquatic environment contamination remains a foremost global public health hazards, and symbolizes a significant reservoir of releasing antibiotic resistant bacteria. The survival of Escherichia coli in aquatic environments serves as a potential reservoir of antibiotic resistance, encompassing but not restricted to a plasmid-mediated quinolone resistance (PMQR) mechanism. The current study aimed to detect the presence of the PMQR-qnrA gene in quinolone-resistant E. coli isolates. Sixty-one waterborne E. coli with known phylogroups/subgroups isolated from the Al-Hillah River in Babylon Province, Iraq, were screened for the phenotypic resistance to third-generation quinolones (levofloxacin and ofloxacin) and were further analysed for the presence of the qnrA gene using polymerase chain reaction (PCR). Fifty-seven (93.4%) of 61 E. coli isolates were levofloxacin-resistant, and 55 (90.2%) were ofloxacin-resistant. Among the 57 quinolone-resistant E. coli, 40 (65.57%) isolates were found to carry the PMQR-qnrA gene. Among the 40 qnrA-positive E. coli, 22 (36.1%) isolates were in phylogroup B2, followed by 8 (13.1%) isolates in phylogroup D, 6 (9.8%) isolates in phylogroup B1, and 4 (6.6%) isolates in phylogroup A. The presence of the PMQR-qnrA gene in E. coli belonging to phylogroup B2 and D reflects the need for routine monitoring of antibiotic resistance genes (ARGs) in the Al-Hillah River.


2021 ◽  
Author(s):  
Tatsuya Nakayama ◽  
Le Thi Hien ◽  
Ngo Thanh Phong ◽  
Doan Nguyen Minh Tran ◽  
Oanh Thi Hoang Nguyen ◽  
...  

Abstract Although the spread of plasmid-mediated antibiotic-resistant bacteria is a public health concern, food contamination with plasmid-mediated antibiotic-resistant Escherichia coli has not been well investigated in Vietnam. The aim of this study was to describe the prevalence of colistin-resistant, carbapenem-resistant and endemic blaCTX−M in extended-spectrum β-lactamase (ESBL)-producing E. coli isolates. Colistin- and carbapenem-resistant ESBL-producing E. coli were isolated from chickens in Vietnam and Japan. The results showed that 52% and 93% of Vietnamese chicken was isolated with colistin-resistant and AmpC/ESBL-producing E. coli, respectively, while 52.7% of Japanese chickens were isolated with AmpC/ESBL-producing E. coli. Carbapenem-resistant E. coli has not been isolated in Vietnam or Japan. Genotyping revealed that colistin-resistant E. coli harboured mcr-1, and most of the AmpC/ESBL-related genes were blaCTX−M−55 and blaCTX−M−65 together with blaTEM in Vietnamese chickens, and blaCMY−2 in Japanese chickens. Multidrug resistance analysis showed that ESBL-producing E. coli isolates were more resistant to quinolones, streptomycin, and chloramphenicol compared with colistin-resistant E. coli isolates from Vietnam, suggesting selection in ESBL-producing E. coli for multiple antibiotic resistance genes. In conclusion, colistin-resistant E. coli was detected in about half of the chicken meat samples, the majority of which were found to harbour mcr-1. The high prevalence of ESBL-producing E. coli has remained constant across the last five years, and the predominant blaCTX−M for ESBL-producing E. coli was found to be blaCTX−M−55 or blaCTX−M−65, with the coexistence of blaTEM in Vietnam. Our results can be implemented in monitoring systems to combat the development of antimicrobial resistance.


2021 ◽  
Vol 12 ◽  
Author(s):  
Lorena Montero ◽  
Jorge Irazabal ◽  
Paul Cardenas ◽  
Jay P. Graham ◽  
Gabriel Trueba

In cities across the globe, the majority of wastewater – that includes drug resistant and pathogenic bacteria among other contaminants – is released into streams untreated. This water is often subsequently used for irrigation of pastures and produce. This use of wastewater-contaminated streams allows antibiotic-resistant bacteria to potentially cycle back to humans through agricultural products. In this study, we investigated the prevalence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolated from produce and irrigation water across 17 provinces of Ecuador. A total of 117 vegetable samples, 119 fruit samples, and 38 irrigation water samples were analyzed. Results showed that 11% of the samples were positive for E. coli including 11 irrigation water samples (29%), and samples of 13 vegetables (11%), and 11 fruits (9%). Among the 165 E. coli isolates cultured, 96 (58%) had the ESBL phenotype, and 58% of ESBL producing E. coli came from irrigation water samples, 11% from vegetables, and 30% from fruits. The blaCTX–M–55, blaCTX–M 65, and blaCTX–M 15 genes were the most frequently found gene associated with the ESBL phenotype and coincided with the blaCTX–M alleles associated with human infections in Ecuador. Three isolates had the mcr-1 gene which is responsible for colistin resistance. This report provides evidence of the potential role of irrigation water in the growing antimicrobial resistance crisis in Ecuador.


Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 850
Author(s):  
Shobha Giri ◽  
Vaishnavi Kudva ◽  
Kalidas Shetty ◽  
Veena Shetty

As the global urban populations increase with rapid migration from rural areas, ready-to-eat (RTE) street foods are posing food safety challenges where street foods are prepared with less structured food safety guidelines in small and roadside outlets. The increased presence of extended-spectrum-β-lactamase (ESBL) producing bacteria in street foods is a significant risk for human health because of its epidemiological significance. Escherichia coli and Klebsiella pneumoniae have become important and dangerous foodborne pathogens globally for their relevance to antibiotic resistance. The present study was undertaken to evaluate the potential burden of antibiotic-resistant E. coli and K. pneumoniae contaminating RTE street foods and to assess the microbiological quality of foods in a typical emerging and growing urban suburb of India where RTE street foods are rapidly establishing with public health implications. A total of 100 RTE food samples were collected of which, 22.88% were E. coli and 27.12% K. pneumoniae. The prevalence of ESBL-producing E. coli and K. pneumoniae was 25.42%, isolated mostly from chutneys, salads, paani puri, and chicken. Antimicrobial resistance was observed towards cefepime (72.9%), imipenem (55.9%), cefotaxime (52.5%), and meropenem (16.9%) with 86.44% of the isolates with MAR index above 0.22. Among β-lactamase encoding genes, blaTEM (40.68%) was the most prevalent followed by blaCTX (32.20%) and blaSHV (10.17%). blaNDM gene was detected in 20.34% of the isolates. This study indicated that contaminated RTE street foods present health risks to consumers and there is a high potential of transferring multi-drug-resistant bacteria from foods to humans and from person to person as pathogens or as commensal residents of the human gut leading to challenges for subsequent therapeutic treatments.


2011 ◽  
Vol 56 (3) ◽  
pp. 1414-1417 ◽  
Author(s):  
Jien-Wei Liu ◽  
Wen-Chien Ko ◽  
Cheng-Hua Huang ◽  
Chun-Hsing Liao ◽  
Chin-Te Lu ◽  
...  

ABSTRACTThe TigecyclineIn VitroSurveillance in Taiwan (TIST) study, initiated in 2006, is a nationwide surveillance program designed to longitudinally monitor thein vitroactivity of tigecycline against commonly encountered drug-resistant bacteria. This study compared thein vitroactivity of tigecycline against 3,014 isolates of clinically important drug-resistant bacteria using the standard broth microdilution and disk diffusion methods. Species studied included methicillin-resistantStaphylococcus aureus(MRSA;n= 759), vancomycin-resistantEnterococcus faecium(VRE;n= 191), extended-spectrum β-lactamase (ESBL)-producingEscherichia coli(n= 602), ESBL-producingKlebsiella pneumoniae(n= 736), andAcinetobacter baumannii(n= 726) that had been collected from patients treated between 2008 and 2010 at 20 hospitals in Taiwan. MICs and inhibition zone diameters were interpreted according to the currently recommended U.S. Food and Drug Administration (FDA) criteria and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. The MIC90values of tigecycline against MRSA, VRE, ESBL-producingE. coli, ESBL-producingK. pneumoniae, andA. baumanniiwere 0.5, 0.125, 0.5, 2, and 8 μg/ml, respectively. The total error rates between the two methods using the FDA criteria were high: 38.4% for ESBL-producingK. pneumoniaeand 33.8% forA. baumannii. Using the EUCAST criteria, the total error rate was also high (54.6%) forA. baumanniiisolates. The total error rates between these two methods were <5% for MRSA, VRE, and ESBL-producingE. coli. For routine susceptibility testing of ESBL-producingK. pneumoniaeandA. baumanniiagainst tigecycline, the broth microdilution method should be used because of the poor correlation of results between these two methods.


2015 ◽  
Vol 7 (3) ◽  
pp. 272-280 ◽  
Author(s):  
Ifeoma Chinyere UGWU ◽  
Madubuike Umunna ANYANWU ◽  
Chidozie Clifford UGWU ◽  
Ogbonna Wilfred UGWUANYI

This study was conducted to isolate generic extended-spectrum β-lactam (ESBL)-resistant enterobacteria from pigs reared in Enugu State Southeast, Nigeria and determine the antibacterial resistance profile of the isolates. Rectal swabs were collected from 190, randomly selected, apparently healthy pigs. Isolation of ESBL-resistant enterobacteria was done using Mac Conkey agar supplemented with 2 µg/ml of cefotaxime. Phenotypic characterization of the isolates to generic level was done following standard biochemical methods. Phenotypic resistance of the isolates to antibacterial agents was determined using the disc diffusion method. Out of 46 ESBL-resistant enterobacterial isolates, 4 (8.7%) were Escherichia coli, 11 (23.9%) were Salmonella species, while 31 (67.4%) were Klebsiella species. Resistance of the Salmonella isolates was 45.5% to ciprofloxacin, 36.4% to ofloxacin and levofloxacin, 9.1% to norfloxacin, amikacin and gentamicin, 27.3% to streptomycin, 72.7% to chloramphenicol and 90.9% to tetracycline. Resistance of the Klebsiella isolates was 93.5% to ampicillin, 12.9% to ciprofloxacin, 19.4% to ofloxacin and levofloxacin, 9.7% to norfloxacin and streptomycin, 64.5% to chloramphenicol and 38.7% to tetracycline. Resistance of the E. coli isolates was 100% to gentamicin, 75% to ampicillin and streptomycin, 50% to ciprofloxacin, norfloxacin, chloramphenicol and tetracycline, and 25% to ofloxacin, levofloxacin and amikacin. All the isolates were resistant to ceftriaxone, cefotaxime, ceftazidime, cefepime, cefpodoxime, amoxicillin/clavulanic acid and aztreonam. Resistance of the isolates to more than 3 classes of antibacterial agents tested was 54.8% for Klebsiella, 90.9% for Salmonella and 100% for E. coli, respectively. This study has shown that pigs reared in Enugu State Southeast, Nigeria, are colonized by ESBL-resistant Enterobactericeae and are potential reservoirs and disseminators of these organisms.


2021 ◽  
Author(s):  
Farhan Yusuf ◽  
Kimberley Gilbride

Bacterial isolates found in aquatic ecosystems often carry antibiotic resistance genes (ARGs). These ARGs are often found on plasmids and transposons, which allows them to be proliferate throughout bacterial communities via horizontal gene transfer (HGT) causing dissemination of multidrug resistance. The increase in antibiotic resistance has raised concerns about the ability to continue to use these drugs to fight infectious diseases. Novel synthetic antibiotics like ciprofloxacin that are not naturally found in the environment were developed to prevent resistances. However, ciprofloxacin resistance has occurred through chromosomal gene mutations of type 2 topoisomerases or by the acquisition of plasmid-mediated quinolone resistances (PMQR). A particular PMQR, qnr genes, encoding for pentapeptide repeat proteins that confer low levels of quinolone resistance and protect DNA gyrase and topoisomerase IV from antibacterial activity. These qnr genes have been identified globally in both clinical and environmental isolates. The aim of this study was to determine the prevalence of ciprofloxacin-resistant bacteria in aquatic environments in the Greater Toronto Area and the potential dissemination of ciprofloxacin resistance. With the selective pressure of ciprofloxacin, we hypothesize that ciprofloxacin-resistant bacteria (CipR) in the environment may carry PMQR mechanisms while the sensitive population (CipS) would not carry PMQR genes. Isolates were tested for resistance to an additional 12 different antibiotics and identified using Sanger sequencing PCR products of the 16S rRNA gene. To determine which genes are responsible for ciprofloxacin resistance, multiplex PCR of associated qnr genes, qnrA, qnrB, and qnrS, was carried out on 202 environmental isolates. Our data demonstrate a similar prevalence of qnr genes was found in CipR (19%) and CipS (14%) populations suggesting that the presence of these genes was not necessarily correlated with the phenotypic resistance to the antibiotic. Furthermore, ciprofloxacinresistant bacteria were found in all locations at similar frequencies suggesting that resistance genes are widespread and could possibly arise through HGT events. Overall, determining the underlying cause and prevalence of ciprofloxacin resistance could help re-establish the effectiveness of these antimicrobial compounds.


2021 ◽  
Author(s):  
Chiara Borsetto ◽  
Sebastien Raguideau ◽  
Emma R Travis ◽  
Dae-Wi Kim ◽  
Do-Hoon Lee ◽  
...  

The continued emergence of bacterial pathogens presenting antimicrobial resistance is widely recognised as a global health threat and recent attention focused on potential environmental reservoirs of antibiotic resistance genes (ARGs). Freshwater environments such as rivers represent a potential hotspot for ARGs and antibiotic resistant bacteria as they are receiving systems for effluent discharges from wastewater treatment plants (WWTPs). Effluent also contains low levels of different antimicrobials including antibiotics and biocides. Sulfonamides are antibacterial chemicals widely used in clinical, veterinary and agricultural settings and are frequently detected in sewage sludge and manure in addition to riverine ecosystems. The impact of such exposure on ARG prevalence and diversity is unknown, so the aim of this study was to investigate the release of a sub-lethal concentration of the sulfonamide compound sulfamethoxazole (SMX) on the river bacterial microbiome using a flume system. This system was a semi-natural in vitro flume using river water (30 L) and sediment with circulation to mimic river flow. A combination of 'omics' approaches were conducted to study the impact of SMX exposure on the microbiomes within the flumes. Metagenomic analysis showed that the addition of low concentrations of SMX (<4 μg L-1) had a limited effect on the bacterial resistome in the water fraction only, with no impact observed in the sediment. Metaproteomics did not show differences in ARGs expression with SMX exposure in water. Overall, the river bacterial community was resilient to short term exposure to sub-lethal concentrations of SMX which mimics the exposure such communities experience downstream of WWTPs throughout the year.


2021 ◽  
Vol 20 (3) ◽  
Author(s):  
Hairul Aini Hamzah ◽  
Rahmatullah Sirat ◽  
Mohammed Imad A. Mustafa Mahmud ◽  
Roesnita Baharudin

 Introduction: Co-resistance to quinolones among extended spectrum β[1]lactamase (ESBL)-producing E. coli commonly occurs in clinical settings. Quinolones act on DNA gyrase and DNA topoisomerase enzymes, which are coded by gyrA and parC genes, thus any mutation to the genes may affect the drug effectiveness. The objective of the study was to characterize gyrA and parC genes in quinolone-resistant E. coli isolates and correlated the mutations with their phenotypic resistance. Materials and Methods: Thirty-two quinolone-resistant (QR) and six quinolone-sensitive (QS) ESBL-E. coli isolates were identified by antibiotic susceptibility and minimum inhibitory concentration tests. Bioinformatics analysis were conducted to study any mutations occurred in the genes and generate their codon compositions. Results: All the QR ESBL-E. coli isolates were identified as multidrug-resistant bacteria. A single point mutation in the quinolone resistance-determining region (QRDR) of gyrA, at codon 83, caused the substitution amino acid Ser83Leu. It is associated with a high level of resistance to nalidixic acid. However, double mutations Ser83Leu and Asp87Asn in the same region were significantly linked to higher levels of resistance to ciprofloxacin. Cumulative point mutations in gyrA and/or in parC were also correlated significantly (p<0.05) to increased resistance to ciprofloxacin. Conclusion: Together, the findings showed that the mutations in gyrA and parC genes handled the institution of intrinsic quinolone resistance in the ESBL-E. coli isolates. Thus, vigilant monitoring for emergence of new mutation in resistance genes may give an insight into dissemination of QR ESBL-E. coli in a particular region.


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