scholarly journals Antibiotic Resistance and Sewage-Associated Marker Genes in Untreated Sewage and a River Characterized During Baseflow and Stormflow

2021 ◽  
Vol 12 ◽  
Author(s):  
Warish Ahmed ◽  
Pradip Gyawali ◽  
Kerry A. Hamilton ◽  
Sayalee Joshi ◽  
David Aster ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Water Samples ◽  
Quantitative Polymerase Chain Reaction ◽  
River System ◽  
Marker Genes ◽  
Chain Reaction ◽  
E Coli ◽  
Untreated Sewage ◽  
Polymerase Chain

Since sewage is a hotspot for antibiotic resistance genes (ARGs), the identification of ARGs in environmental waters impacted by sewage, and their correlation to fecal indicators, is necessary to implement management strategies. In this study, sewage treatment plant (STP) influent samples were collected and analyzed using quantitative polymerase chain reaction (qPCR) to investigate the abundance and correlations between sewage-associated markers (i.e., Bacteroides HF183, Lachnospiraceae Lachno3, crAssphage) and ARGs indicating resistance to nine antibiotics (belonging to aminoglycosides, beta-lactams, sulfonamides, macrolides, and tetracyclines). All ARGs, except blaVIM, and sewage-associated marker genes were always detected in untreated sewage, and ermF and sul1 were detected in the greatest abundances. intl1 was also highly abundant in untreated sewage samples. Significant correlations were identified between sewage-associated marker genes, ARGs and the intl1 in untreated sewage (τ = 0.488, p = 0.0125). Of the three sewage-associated marker genes, the BIO-ENV procedure identified that HF183 alone best maximized correlations to ARGs and intl1 (τ = 0.590). Additionally, grab samples were collected from peri-urban and urban sites along the Brisbane River system during base and stormflow conditions, and analyzed for Escherichia coli, ARGs, the intl1, and sewage-associated marker genes using quantitative polymerase chain reaction (qPCR). Significant correlations were identified between E. coli, ARGs, and intl1 (τ = 0.0893, p = 0.0032), as well as with sewage-associated marker genes in water samples from the Brisbane River system (τ = 0.3229, p = 0.0001). Of the sewage-associated marker genes and E. coli, the BIO-ENV procedure identified that crAssphage alone maximized correlations with ARGs and intl1 in river samples (τ = 0.4148). Significant differences in E. coli, ARGs, intl1, and sewage-associated marker genes, and by flow condition (i.e., base vs. storm), and site types (peri-urban vs. urban) combined were identified (R = 0.3668, p = 0.0001), where percent dissimilarities between the multi-factorial groups ranged between 20.8 and 11.2%. Results from this study suggest increased levels of certain ARGs and sewage-associated marker genes in stormflow river water samples compared to base flow conditions. E. coli, HF183 and crAssphage may serve as potential indicators of sewage-derived ARGs under stormflow conditions, and this merits further investigation. Data presented in this study will be valuable to water quality managers to understand the links between sewage pollution and ARGs in urban environments.

scholarly journals Use of Fluorescence Quantitative Polymerase Chain Reaction (PCR) for the Detection of Escherichia coli Adhesion to Pig Intestinal Epithelial Cells

2016 ◽  
Vol 19 (3) ◽  
pp. 619-625 ◽  
Author(s):  
C.H. Dai ◽  
L.N. Gan ◽  
W.U. Qin ◽  
C. Zi ◽  
G.Q. Zhu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Relative Number ◽  
Intestinal Epithelial ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain

AbstractAn efficient and accurate method to testEscherichia coli(E. coli) adhesion to intestinal epithelial cells will contribute to the study of bacterial pathogenesis and the function of genes that encode receptors related to adhesion. This study used the quantitative real-time polymerase chain reaction (qPCR) method. qPCR primers were designed from thePILINgene ofE. coliF18ab, F18ac, and K88ac, and the pig β-ACTINgene. Total deoxyribonucleic acid (DNA) fromE. coliand intestinal epithelial cells (IPEC-J2 cells) were used as templates for qPCR. The 2−ΔΔCtformula was used to calculate the relative number of bacteria in cultures of different areas. We found that the relative numbers of F18ab, F18ac, and K88ac that adhered to IPEC-J2 cells did not differ significantly in 6-, 12-, and 24-well culture plates. This finding indicated that there was no relationship between the relative adhesion number ofE. coliand the area of cells, so the method of qPCR could accurately test the relative number ofE. coli. This study provided a convenient and reliable testing method for experiments involvingE. coliadhesion, and also provided innovative ideas for similar detection methods.

scholarly journals Diverse Escherichia coli pathovars of phylogroups B2 and D isolated from animals in Tunisia

2017 ◽  
Vol 11 (07) ◽  
pp. 549-556 ◽  
Author(s):  
Hajer Kilani ◽  
Mohamed Salah Abbassi ◽  
Sana Ferjani ◽  
Rakia Ben Salem ◽  
Riadh Mansouri ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Virulence Genes ◽  
Chain Reaction ◽  
E Coli ◽  
Class 1 ◽  
Genes Encoding ◽  
Polymerase Chain ◽  
Relationship Of

Introduction: The virulent Escherichia coli strains responsible for extraintestinal infections were mainly belonged to B2 and D phylogroups. However, no past studies have determinate via the presence of virulence genes the frequency of E. coli pathovars recovered from animals housed in farms in Tunisia. The aims of this study were to investigate 26 E. coli isolated from healthy and diarrheic animals and to determinate via the presence of virulence genes the frequency of pathovars. Methodology: Twenty-six E. coli isolates of phylogroups B2 (n = 14), B22 (n = 9), B23 (n = 5), and D2 (n = 12) were characterized. Genes encoding virulence factors (fimH,eaeA,aggC,papC, papG allele III, hlyA, east1, cnf1, exhA,stx1, stx2, iutA, fyuA, ibeA,and ipaH), and antibiotic resistance as well as class 1 and 2 integrons were searched by polymerase chain reaction (PCR). The genetic relationship of isolates was done by PFGE. Results: According to the occurrence of specific genes the 26 isolates were classified as:9 EAEC, 2 EHEC, 4 UPEC, 3 EPEC/EHEC and 1 NTEC. Therefore, 2 Ex-PEC and 5 APEC were presented amongst our strains. Some isolates (12) were clonal and the remaining was unrelated. Conclusions: Higher diversity of pathovars which carried diverse combinations of virulence genes in healthy isolates. In addition, it seems that the infections were caused by different mechanisms.

scholarly journals Plant-Derived Antimicrobials ReduceE. coliO157:H7 Virulence Factors Critical for Colonization in Cattle Gastrointestinal TractIn Vitro

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Sangeetha Ananda Baskaran ◽  
Kumar Venkitanarayanan
Keyword(s):  
Polymerase Chain Reaction ◽  
Epithelial Cells ◽  
Rumen Fluid ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
E Coli ◽  
Intestinal Contents ◽  
Polymerase Chain

This study investigated the effect of subinhibitory concentrations (SIC) of five plant-derived antimicrobials (PDAs), namely, trans cinnamaldehyde, eugenol, carvacrol, thymol, andβ-resorcylic acid, onE. coliO157:H7 (EHEC) attachment and invasion of cultured bovine colonic (CO) and rectoanal junction (RAJ) epithelial cells. In addition, PDAs’ effect on EHEC genes critical for colonization of cattle gastrointestinal tract (CGIT) was determined in bovine rumen fluid (RF) and intestinal contents (BICs). Primary bovine CO and RAJ epithelial cells were established and were separately inoculated with three EHEC strains with or without (control) SIC of each PDA. Following incubation, EHEC that attached and invaded the cells were determined. Furthermore, the expression of EHEC genes critical for colonization in cattle was investigated using real-time, quantitative polymerase chain reaction in RF and BICs. All the PDAs decreased EHEC invasion of CO and RAJ epithelial cells (P<0.05). The PDAs also downregulated (P<0.05) the expression of EHEC genes critical for colonization in CGIT. Results suggest that the PDAs could potentially be used to control EHEC colonization in cattle; however follow-upin vivostudies in cattle are warranted.

scholarly journals Development of an Escherichia coli K12-specific quantitative polymerase chain reaction assay and DNA isolation suited to biofilms associated with iron drinking water pipe corrosion products

2014 ◽  
Vol 12 (4) ◽  
pp. 763-771 ◽  
Author(s):  
Jingrang Lu ◽  
Tammie L. Gerke ◽  
Helen Y. Buse ◽  
Nicholas J. Ashbolt
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Drinking Water ◽  
Polymerase Chain Reaction Assay ◽  
Quantitative Polymerase Chain Reaction ◽  
The Novel ◽  
Water Pipe ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain

A quantitative polymerase chain reaction assay (115 bp amplicon) specific to Escherichia coli K12 with an ABITM internal control was developed based on sequence data encoding the rfb gene cluster. Assay specificity was evaluated using three E. coli K12 strains (ATCC W3110, MG1655 & DH1), 24 non-K12 E. coli and 23 bacterial genera. The biofilm detection limit was 103 colony-forming units (CFU) E. coli K12 mL−1, but required a modified protocol, which included a bio-blocker Pseudomonas aeruginosa with ethylenediaminetetraacetic acid buffered to pH 5 prior to cell lysis/DNA extraction. The novel protocol yielded the same sensitivity for drinking water biofilms associated with Fe3O4 (magnetite)-coated SiO2 (quartz) grains and biofilm-surface iron corrosion products from a drinking water distribution system. The novel DNA extraction protocol and specific E. coli K12 assay are sensitive and robust enough for detection and quantification within iron drinking water pipe biofilms, and are particularly well suited for studying enteric bacterial interactions within biofilms.

scholarly journals Antibiotic Resistance ofEscherichia coliSerotypes from Cochin Estuary

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Divya P. Sukumaran ◽  
Srinivasan Durairaj ◽  
Mohamed Hatha Abdulla
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Antimicrobial Agents ◽  
Antibiotic Sensitivity ◽  
Cochin Estuary ◽  
Chain Reaction ◽  
Monthly Basis ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Polymerase Chain

This study aimed at detecting the prevalence of antibiotic-resistant serotypes ofEscherichia coliin Cochin estuary, India.E. colistrains were isolated during the period January 2010–December 2011 from five different stations set at Cochin estuary. Water samples from five different stations in Cochin estuary were collected on a monthly basis for a period of two years. Isolates were serotyped, antibiogram-phenotyped for twelve antimicrobial agents, and genotyped by polymerase chain reaction foruidgene that codes forβ-D-glucuronidase. TheseE. colistrains from Cochin estuary were tested against twelve antibiotics to determine the prevalence of multiple antibiotic resistance among them. The results revealed that more than 53.33% of the isolates were multiple antibiotic resistant. Thirteen isolates showed resistance to sulphonamides and two of them contained thesul1 gene. Class 1 integrons were detected in twoE. colistrains which were resistant to more than seven antibiotics. In the present study, O serotyping, antibiotic sensitivity, and polymerase chain reaction were employed with the purpose of establishing the present distribution of multiple antibiotic-resistant serotypes, associated withE. coliisolated from different parts of Cochin estuary.

scholarly journals Identification, characterization and molecular epidemiology of Escherichia coli isolated from lamb and goat kids with diarrhoea

2013 ◽  
Vol 82 (4) ◽  
pp. 357-362 ◽  
Author(s):  
Süheyla Türkyılmaz ◽  
Seza Eskiizmirliler ◽  
Serra Tunaligil ◽  
Bulent Bozdogan
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Small Ruminants ◽  
Multi Drug Resistance ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
E Coli ◽  
Neonatal Diarrhoea ◽  
Polymerase Chain

Neonatal diarrhoea is a serious health problem on commercial farms. EnterovirulentEscherichia coliis a significant aetiological agent of neonatal diarrhoea. In this work, identification and classification ofE. coliisolates obtained from lambs and goat kids with diarrhoea were studied along with antibiotic resistance and clonal relationships of enterovirulent strains. A total of 107E. colistrains isolated from animals on 43 farms were investigated. Specific virulence genes were determined by multiplex and uniplex polymerase chain reaction. Testing of antibiotic susceptibility was carried out by the Vitek II compact system. The relationship ofE. coliisolates was determined by enterobacterial repetitive intergenic consensus polymerase chain reaction. A total of 39 (36.4%) enterovirulentE. colistrains were identified and of this 19 (48.7%) were shiga toxigenic, 12 (30.8%) enterotoxigenic and 8 (20.5%) enteropathogenic. Three isolates (7.7%) were found to be positive for extended spectrum beta lactamase; 10 (25.6%) isolates showed multi-drug resistance to antimicrobials. A total of 28 types were detected by enterobacterial repetitive intergenic consensus polymerase chain reaction. Twenty strains had distinct types while 5 types were common for 2 strains and 3 types were common for 3 strains. This is the first current determination of types, clonality and antibiotic resistance of enterovirulentE. coliisolated from small ruminants with diarrhoea. The results of this study showed that the rates of shiga toxigenic, enterotoxigenic and enteropathogenic isolates ofE. coliare high in the western part of Turkey. Although these isolates were not clonal, presence of multidrug resistant isolates may cause public health problems.

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