scholarly journals AMR Escherichia coli and its temporal and spatial variability within the aquatic environment

2020 ◽  
Vol 2 (7A) ◽  
Author(s):  
Amelia Odell ◽  
Paul Eady ◽  
Ronald Dixon
Keyword(s):  
Escherichia Coli ◽  
Aquatic Environment ◽  
Spatial Dynamics ◽  
Pcr Analysis ◽  
Terrestrial Environment ◽  
E Coli ◽  
Wwtp Effluent ◽  
Temporal And Spatial Variability ◽  
Temporal And Spatial ◽  
Genotypic Identification

Phenotypic and genotypic identification methods have been used to determine the temporal and spatial dynamics of AMR-Escherichia coli in a mainly rural watercourse that receives WWTP-effluent compared to a parallel river which does not. We aimed to investigate the incidence of plasmid-mediated mcr-1and β-lactamase-genes in E. coli recovered from both water and Asellus aquaticus samples throughout two-calendar-years. Samples of the water and A. aquaticus were recovered from the relevant locations each month. CHROMagar ESBL agar was used throughout to isolate and identify ESBL-E. coli. The presence of AMR-genes was confirmed using the ‘BSAC’ antibiotic-disk-synergy method and PCR analysis to confirm the presence of mcr-1and ESBL-genes. The CHROMagar ESBL agar was found to be 99.7% (n=578) accurate when confirmed with a PCR analysis of the ESBL-genes. Seventy-six-point-six percent (n=449) of the isolated ESBL-E. coli were correctly identified as ESBL-producing organisms using the ‘BSAC’ method. Interestingly 61.9% (n=358) of the ESBL-E. coli were also found to carry the mcr-1 gene. Our data shows that AMR levels were highest at the WWTP-effluent throughout the two-years for both water and A. aquaticus samples. The incidence of AMR-E. coli 1km downstream of the effluent discharge was equivalent to the parallel river sites, suggesting that the dispersal of AMR from the WWTP-effluent is limited, although AMR-E. coli were found in relatively high numbers at the WWTP-effluent. We argue that the presence of AMR in the freshwater invertebrate A. aquaticus could represent an important route by which AMR can spread from the aquatic environment to the terrestrial environment.

scholarly journals Molecular surveillance of shiga toxigenic Escherichia coli in selected beef abattoirs in Osun State Nigeria

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Femi Ayoade ◽  
Judith Oguzie ◽  
Philomena Eromon ◽  
Omolola E. Omotosho ◽  
Tosin Ogunbiyi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Sequence Analysis ◽  
Block Design ◽  
Latex Agglutination ◽  
Pcr Analysis ◽  
Accession Number ◽  
E Coli ◽  
Representative Isolate ◽  
Randomized Block Design ◽  
Toxigenic Strains

AbstractShiga toxigenic strains of E. coli (STEC) known to be etiological agents for diarrhea were screened for their incidence/occurrence in selected abattoirs sources in Osogbo metropolis of Osun State, Nigeria using a randomized block design. Samples were plated directly on selective and differential media and E. coli isolates. Multiplex PCR analysis was used to screen for the presence of specific virulence factors. These were confirmed serologically as non-O157 STEC using latex agglutination serotyping kit. Sequence analysis of PCR products was performed on a representative isolate showing the highest combination of virulence genes using the 16S gene for identification purposes only. Results showed that the average cfu/cm2 was significantly lower in the samples collected at Sekona-2 slaughter slab compared with those collected at Al-maleek batch abattoir and Sekona-1 slaughter slab in ascending order at P = 0.03. Moreover, the average cfu/cm2E. coli in samples collected from butchering knife was significantly lower when compared with that of the workers’ hand (P = 0.047) and slaughtering floor (P = 0.047) but not with the slaughter table (P = 0.98) and effluent water from the abattoir house (P = 0.39). These data suggest that the abattoir type may not be as important in the prevalence and spread of STEC as the hygiene practices of the workers. Sequence analysis of a representative isolate showed 100% coverage and 96.46% percentage identity with Escherichia coli O113:H21 (GenBank Accession number: CP031892.1) strain from Canada. This sequence was subsequently submitted to GenBank with accession number MW463885. From evolutionary analyses, the strain from Nigeria, sequenced in this study, is evolutionarily distant when compared with the publicly available sequences from Nigeria. Although no case of E. coli O157 was found within the study area, percent occurrence of non-O157 STEC as high as 46.3% at some of the sampled sites is worrisome and requires regulatory interventions in ensuring hygienic practices at the abattoirs within the study area.

scholarly journals Impact of Ceftiofur Injection on Gut Microbiota and Escherichia coli Resistance in Pigs

2015 ◽  
Vol 59 (9) ◽  
pp. 5171-5180 ◽  
Author(s):  
M. A. Fleury ◽  
G. Mourand ◽  
E. Jouy ◽  
F. Touzain ◽  
L. Le Devendec ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gut Microbiota ◽  
Treated Group ◽  
Conjugative Plasmid ◽  
Health Concern ◽  
Pcr Analysis ◽  
Contamination Level ◽  
Content Type ◽  
E Coli ◽  
The Impact

ABSTRACTResistance to extended-spectrum cephalosporins (ESCs) is an important health concern. Here, we studied the impact of the administration of a long-acting form of ceftiofur on the pig gut microbiota and ESC resistance inEscherichia coli. Pigs were orally inoculated with an ESC-resistantE. coliM63 strain harboring a conjugative plasmid carrying a gene conferring resistance,blaCTX-M-1. On the same day, they were given or not a unique injection of ceftiofur. Fecal microbiota were studied using quantitative PCR analysis of the main bacterial groups and quantification of short-chain fatty acids.E. coliand ESC-resistantE. coliwere determined by culture methods, and the ESC-resistantE. coliisolates were characterized. The copies of theblaCTX-M-1gene were quantified. After ceftiofur injection, the main change in gut microbiota was the significant but transitory decrease in theE. colipopulation. Acetate and butyrate levels were significantly lower in the treated group. In all inoculated groups,E. coliM63 persisted in most pigs, and theblaCTX-M-1gene was transferred to otherE. coli. Culture and PCR results showed that the ceftiofur-treated group shed significantly more resistant strains 1 and 3 days after ESC injection. Thereafter, on most dates, there were no differences between the groups, but notably, one pig in the nontreated group regularly excreted very high numbers of ESC-resistantE. coli, probably leading to a higher contamination level in its pen. In conclusion, the use of ESCs, and also the presence of high-shedding animals, are important features in the spread of ESC resistance.

Multiplex Real-Time PCR Detection of Heat-Labile and Heat-Stable Toxin Genes in Enterotoxigenic Escherichia coli †

2006 ◽  
Vol 69 (2) ◽  
pp. 412-416 ◽  
Author(s):  
MICHAEL A. GRANT ◽  
JINXIN HU ◽  
KAREN C. JINNEMAN
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Real Time Pcr ◽  
Membrane Filtration ◽  
Enterotoxigenic Escherichia Coli ◽  
Pcr Analysis ◽  
E Coli ◽  
Toxin Genes ◽  
Heat Stable ◽  
Heat Labile

A multiplex real-time PCR method was developed for detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli. Approximately 10 CFU per reaction mixture could be detected in rinsates from produce samples. Several foods representative of varieties previously shown to have caused enterotoxigenic E. coli outbreaks were spiked and enriched for 4 or 6 h. Both heat-labile and heat-stable toxin genes could be detected in the foods tested, with the exception of hot sauce, with threshold cycle values ranging from 25.2 to 41.1. A procedure using membrane filtration which would allow enumeration of the enterotoxigenic E. coli population in a food sample in less than 28 h by real-time PCR analysis of colonies picked from media highly selective for E. coli was also developed.

scholarly journals Unexpected Stress-Reducing Effect of PhaP, a Poly(3-Hydroxybutyrate) Granule-Associated Protein, in Escherichia coli

2011 ◽  
Vol 77 (18) ◽  
pp. 6622-6629 ◽  
Author(s):  
Alejandra de Almeida ◽  
Mariela V. Catone ◽  
Virgil A. Rhodius ◽  
Carol A. Gross ◽  
M. Julia Pettinari
Keyword(s):  
Escherichia Coli ◽  
Heat Shock ◽  
Stress Proteins ◽  
Protective Role ◽  
Pcr Analysis ◽  
Content Type ◽  
E Coli ◽  
Protein Levels ◽  
Stress Related Genes ◽  
Heat Stress Proteins

ABSTRACTPhasins (PhaP) are proteins normally associated with granules of poly(3-hydroxybutyrate) (PHB), a biodegradable polymer accumulated by many bacteria as a reserve molecule. These proteins enhance growth and polymer production in natural and recombinant PHB producers. It has been shown that the production of PHB causes stress in recombinantEscherichia coli, revealed by an increase in the concentrations of several heat stress proteins. In this work, quantitative reverse transcription (qRT)-PCR analysis was used to study the effect of PHB accumulation, and that of PhaP fromAzotobactersp. strain FA8, on the expression of stress-related genes in PHB-producingE. coli. While PHB accumulation was found to increase the transcription ofdnaKandibpA, the expression of these genes and ofgroES,groEL,rpoH,dps, andyfiDwas reduced, when PhaP was coexpressed, to levels even lower than those detected in the non-PHB-accumulating control. These results demonstrated the protective role of PhaP in PHB-synthesizingE. coliand linked the effects of the protein to the expression of stress-related genes, especiallyibpA. The effect of PhaP was also analyzed in non-PHB-synthesizing strains, showing that expression of this heterologous protein has an unexpected protective effect inE. coli, under both normal and stress conditions, resulting in increased growth and higher resistance to both heat shock and superoxide stress by paraquat. In addition, PhaP expression was shown to reduce RpoH protein levels during heat shock, probably by reducing or titrating the levels of misfolded proteins.

scholarly journals Sab, a Novel Autotransporter of Locus of Enterocyte Effacement-Negative Shiga-Toxigenic Escherichia coli O113:H21, Contributes to Adherence and Biofilm Formation

2009 ◽  
Vol 77 (8) ◽  
pp. 3234-3243 ◽  
Author(s):  
Sylvia Herold ◽  
James C. Paton ◽  
Adrienne W. Paton
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Gastrointestinal Disease ◽  
Pcr Analysis ◽  
Systemic Complications ◽  
E Coli ◽  
Acid Protein ◽  
Life Threatening ◽  
Uremic Syndrome ◽  
Enterocyte Effacement

ABSTRACT Shiga-toxigenic Escherichia coli (STEC) strains cause serious gastrointestinal disease, which can lead to potentially life-threatening systemic complications such as hemolytic-uremic syndrome. Although the production of Shiga toxin has been considered to be the main virulence trait of STEC for many years, the capacity to colonize the host intestinal epithelium is a crucial step in pathogenesis. In this study, we have characterized a novel megaplasmid-encoded outer membrane protein in locus of enterocyte effacement (LEE)-negative O113:H21 STEC strain 98NK2, termed Sab (for STEC autotransporter [AT] contributing to biofilm formation). The 4,296-bp sab gene encodes a 1,431-amino-acid protein with the features of members of the AT protein family. When expressed in E. coli JM109, Sab contributed to the diffuse adherence to human epithelial (HEp-2) cells and promoted biofilm formation on polystyrene surfaces. A 98NK2 sab deletion mutant was also defective in biofilm formation relative to its otherwise isogenic wild-type parent, and this was complemented by transformation with a sab-carrying plasmid. Interestingly, an unrelated O113:H21 STEC isolate that had a naturally occurring deletion in sab was similarly defective in biofilm formation. PCR analysis indicated that sab is present in LEE-negative STEC strains belonging to serotypes/groups O113:H21, O23, and O82:H8. These findings raise the possibility that Sab may contribute to colonization in a subset of LEE-negative STEC strains.

scholarly journals Extended-spectrum beta-lactamase (ESBL)-positive Escherichia coli presence in urban aquatic environments in Kanpur, India

2020 ◽  
Vol 18 (5) ◽  
pp. 849-854
Author(s):  
Ann Johnson ◽  
Olivia Ginn ◽  
Aaron Bivins ◽  
Lucas Rocha-Melogno ◽  
Sachchida Nand Tripathi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Aquatic Environment ◽  
Significant Risk ◽  
Uttar Pradesh ◽  
Environmental Water ◽  
Antibiotic Consumption ◽  
Beta Lactamase ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum

Abstract In India, high rates of antibiotic consumption and poor sanitation infrastructure combine to pose a significant risk to the public through the environmental transmission of antimicrobial resistance (AMR). The WHO has declared extended-spectrum beta-lactamase (ESBL)-positive Escherichia coli a key indicator for the surveillance of AMR worldwide. In the current study, we measured the prevalence of AMR bacteria in an urban aquatic environment in India by detecting metabolically active ESBL-positive E. coli. Water samples were collected in duplicate from 16 representative environmental water sources including open canals, drains, and rivers around Kanpur, Uttar Pradesh. We detected culturable E. coli in environmental water at 11 (69%) of the sites. Out of the 11 sites that were positive for culturable E. coli, ESBL-producing E. coli was observed at 7 (64%). The prevalence of ESBL-producing E. coli detected in the urban aquatic environment suggests a threat of AMR bacteria to this region.

scholarly journals Threats to aquatic ecosystems caused by antibiotic-resistant isolate of Escherichia coli from sewage

2016 ◽  
Vol 48 (4) ◽  
pp. 341-351
Author(s):  
Magdalena Frąk ◽  
Zuzanna Kozerska
Keyword(s):  
Escherichia Coli ◽  
Aquatic Environment ◽  
Aquatic Ecosystems ◽  
Resistant Isolate ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Low Concentrations ◽  
Resistance To Penicillin

Abstract The occurrence of Escherichia coli isolate resistant to penicillin and streptomycin in sewage discharged into the environment was tested. Thirty three Escherichia coli isolate were isolated from sewage samples showed different susceptibility to tested antibiotics. All tested isolate show higher resistance to penicillin than streptomycin. Twenty four tested E. coli isolate showed resistance only to low concentrations of penicillin. Five E. coli isolate showed resistance to higher concentrations of penicillin as well (120 μg·dm−3). Five E. coli isolate showed resistance to penicillin and streptomycin. Discharging sewage that contains bacteria isolate resistant to antibiotics into the aquatic environment causes their spreading and increases threats to aquatic ecosystems.

scholarly journals Genomic Insights of Multidrug-Resistant Escherichia coli From Wastewater Sources and Their Association With Clinical Pathogens in South Africa

2021 ◽  
Vol 8 ◽  
Author(s):  
Joshua Mbanga ◽  
Daniel G. Amoako ◽  
Akebe L. K. Abia ◽  
Mushal Allam ◽  
Arshad Ismail ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Treatment Plant ◽  
Multidrug Resistant ◽  
Genomic Diversity ◽  
Health Concern ◽  
Comparative Genomic ◽  
Phylogenomic Analysis ◽  
E Coli ◽  
Wwtp Effluent

There is limited information on the comparative genomic diversity of antibiotic-resistant Escherichia coli from wastewater. We sought to characterize environmental E. coli isolates belonging to various pathotypes obtained from a wastewater treatment plant (WWTP) and its receiving waters using whole-genome sequencing (WGS) and an array of bioinformatics tools to elucidate the resistomes, virulomes, mobilomes, clonality, and phylogenies. Twelve multidrug-resistant (MDR) diarrheagenic E. coli isolates were obtained from the final effluent of a WWTP, and the receiving river upstream and downstream of the WWTP were sequenced on an Illumina MiSeq machine. The multilocus sequence typing (MLST) analysis revealed that multiple sequence types (STs), the most common of which was ST69 (n = 4) and ST10 (n = 2), followed by singletons belonging to ST372, ST101, ST569, ST218, and ST200. One isolate was assigned to a novel ST ST11351. A total of 66.7% isolates were positive for β-lactamase genes with 58.3% harboring the blaTEM1B gene and a single isolate the blaCTX−M−14 and blaCTX−M−55 extended-spectrum β-lactamase (ESBL) genes. One isolate was positive for the mcr-9 mobilized colistin resistance gene. Most antibiotic resistance genes (ARGs) were associated with mobile genetic support: class 1 integrons (In22, In54, In191, and In369), insertion sequences (ISs), and/or transposons (Tn402 or Tn21). A total of 31 virulence genes were identified across the study isolates, including those responsible for adhesion (lpfA, iha, and aggR), immunity (air, gad, and iss), and toxins (senB, vat, astA, and sat). The virulence genes were mostly associated with IS (IS1, IS3, IS91, IS66, IS630, and IS481) or prophages. Co-resistance to heavy metal/biocide, antibiotics were evident in several isolates. The phylogenomic analysis with South African E. coli isolates from different sources (animals, birds, and humans) revealed that isolates from this study mostly clustered with clinical isolates. Phylogenetics linked with metadata revealed that isolates did not cluster according to source but according to ST. The occurrence of pathogenic and MDR isolates in the WWTP effluent and the associated river is a public health concern.

Prevalence and Numbers of Escherichia coli O157 on Bovine Hides at a Beef Slaughter Plant

2005 ◽  
Vol 68 (4) ◽  
pp. 660-665 ◽  
Author(s):  
S. B. O[apos]BRIEN ◽  
G. DUFFY ◽  
E. CARNEY ◽  
J. J. SHERIDAN ◽  
D. A. McDOWELL ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Early Stage ◽  
Immunomagnetic Separation ◽  
Pcr Analysis ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Hemolysin Gene ◽  
Peptone Water ◽  
Encoding Gene ◽  
Genetic Profiles

In this study, we investigated the prevalence and numbers of Escherichia coli O157 on bovine hides. Samples (n = 1,500) were collected over a 17-month period (30 samples per week) by sponge swabbing approximately 122-cm2 areas of the bovine rump of slaughtered cattle at an early stage of carcass processing (first legging). Sponge samples (n = 1,500) were stomached in buffered peptone water supplemented with novobiocin, directly plated on sorbitol MacConkey with Cefixime tellurite (SMAC-CT), enriched for 24 h, extracted by immunomagnetic separation, and plated onto SMAC-CT agar. Presumptive E. coli O157 colonies from SMAC-CT plates were confirmed by PCR for the presence of eaeA, hlyA, fliCh7, vt1, vt2, and portions of the rfb (O-antigen encoding) region of E. coli O157. Overall, E. coli O157 was recovered from 109 samples (7.3%) at concentrations ranging from less than 0.13 to 4.24 log CFU/100 cm2. PCR analysis revealed a wide diversity of genetic profiles among recovered isolates of verocytotoxigenic E. coli. Of the isolates recovered, 99 of 109 contained the attaching and effacing gene (eaeA) and the hemolysin gene (hlyA), and 78 of 109 had the flagellar H7 antigen–encoding gene (fliCh7). Only 6 of 109 isolates contained both verotoxin-producing genes (vt1 and vt2); 91 of 109 contained the vt2 gene only, whereas 1 of 109 contained the vt1 gene only. The remaining 11 of 109 contained neither vt1 nor vt2.

scholarly journals Inversions over the Terminus Region in Salmonella and Escherichia coli: IS200s as the Sites of Homologous Recombination Inverting the Chromosome of Salmonella enterica Serovar Typhi

2002 ◽  
Vol 184 (22) ◽  
pp. 6190-6197 ◽  
Author(s):  
Suneetha Alokam ◽  
Shu-Lin Liu ◽  
Kamal Said ◽  
Kenneth E. Sanderson
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enterica ◽  
Genome Structure ◽  
Enteric Bacteria ◽  
Genomic Rearrangements ◽  
Pcr Analysis ◽  
E Coli ◽  
Salmonella Enterica Serovar Typhi ◽  
Serovar Typhimurium ◽  
Type Strains

ABSTRACT Genomic rearrangements (duplications and inversions) in enteric bacteria such as Salmonella enterica serovar Typhimurium LT2 and Escherichia coli K12 are frequent (10−3 to 10−5) in culture, but in wild-type strains these genomic rearrangements seldom survive. However, inversions commonly survive in the terminus of replication (TER) region, where bidirectional DNA replication terminates; nucleotide sequences from S. enterica serovar Typhimurium LT2, S. enterica serovar Typhi CT18, E. coli K12, and E. coli O157:H7 revealed genomic inversions spanning the TER region. Assuming that S. enterica serovar Typhimurium LT2 represents the ancestral genome structure, we found an inversion of 556 kb in serovar Typhi CT18 between two of the 25 IS200 elements and an inversion of about 700 kb in E. coli K12 and E. coli O157:H7. In addition, there is another inversion of 500 kb in E. coli O157:H7 compared with E. coli K12. PCR analysis confirmed that all S. enterica serovar Typhi strains tested, but not strains of other Salmonella serovars, have an inversion at the exact site of the IS200 insertions. We conclude that inversions of the TER region survive because they do not significantly change replication balance or because they are part of the compensating mechanisms to regain chromosome balance after it is disrupted by insertions, deletions, or other inversions.

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