scholarly journals Temporal Appraisal and Molecular Characterization of Escherichia coli from Oyun River, Kwara State, Nigeria

2020 ◽  
pp. 56-63
Author(s):  
Olatunji Matthew Kolawole ◽  
Oluwasanmi Anuoluwapo Adeyemi
Keyword(s):  
Escherichia Coli ◽  
Molecular Characterization ◽  
Bacterial Load ◽  
Disease Outbreaks ◽  
Coli Strain ◽  
Microbial Count ◽  
Mean Values ◽  
E Coli ◽  
Risk Variants ◽  
Different Strains

Introduction: Escherichia coli, an indicator of feacal contamination has been proven to be the cause of several disease outbreaks in countries and continents around the world. Aim: To determine the genotypic variants of Escherichia coli present in Oyun River and provide information regarding the high-risk variants of E. coli in Oyun River. Study Design: The study cuts across the two seasons of Nigeria’s tropical climate weather, being the peak of the Harmattan season and the onset of the Rainy season. Three sampling sites (Jimba Oja; Unilorin Dam and Oyun in Ilorin, Kwara State) along the River course were examined for three months (February – April). Methodology: Heterotrophic counts, coli form counts and molecular characterization via PCR   using 16 sRNA primers, of water samples were done using standard microbiological and molecular methods. Results: Bacteriological results showed monthly mean values of microbial counts, ranged from 23.5 x 106 – 45.17 x 106 cfu/mL and total coli form count ranged from 53 cfu/100 mL to 256 cfu/100 mLs, both of which exceed the WHO standards of 100 cfu/mL for total microbial count and <1 cfu/100 mLs for total coliforms. A total of forty-eight coliforms were isolated, thirty two of which were Escherichia coli. Sequencing and BLAST analysis of eleven of the isolates using NCBI’s online database revealed five different strains. They include: Escherichia coli FAP1 genome (9.1%); Escherichia coli strain ST2747 (54.5%); Escherichia coli strain EADK4 (9.1%); Escherichia coli strain ST540 (18.2%) and Escherichia coli strain NCM3722 (9.1%). Correlation of results with previous studies showed that most of the strains identified were pathogenic. The E. coli strains isolated, coupled with the bacterial load, coliform count and some physicochemical parameters of Oyun River makes it unsafe for public consumption if not treated. Efforts therefore should be made to treat the water before use, while making frantic efforts to prevent further contamination of Oyun River.

scholarly journals Transmission of Escherichia coli from Manure to Root Zones of Field-Grown Lettuce and Leek Plants

2021 ◽  
Vol 9 (11) ◽  
pp. 2289
Author(s):  
Leo van Overbeek ◽  
Marie Duhamel ◽  
Stefan Aanstoot ◽  
Carin Lombaers van der Plas ◽  
Els Nijhuis ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Plant Root ◽  
Disease Outbreaks ◽  
Coli Strain ◽  
E Coli ◽  
Root Zones ◽  
Food Borne ◽  
Metagenome Library ◽  
Fresh Vegetables ◽  
Vegetables And Fruits

Pathogenic Escherichia coli strains are responsible for food-borne disease outbreaks upon consumption of fresh vegetables and fruits. The aim of this study was to establish the transmission route of E. coli strain 0611, as proxy for human pathogenic E. coli, via manure, soil and plant root zones to the above-soil plant compartments. The ecological behavior of the introduced strain was established by making use of a combination of cultivation-based and molecular targeted and untargeted approaches. Strain 0611 CFUs and specific molecular targets were detected in the root zones of lettuce and leek plants, even up to 272 days after planting in the case of leek plants. However, no strain 0611 colonies were detected in leek leaves, and only in one occasion a single colony was found in lettuce leaves. Therefore, it was concluded that transmission of E. coli via manure is not the principal contamination route to the edible parts of both plant species grown under field conditions in this study. Strain 0611 was shown to accumulate in root zones of both species and metagenomic reads of this strain were retrieved from the lettuce rhizosphere soil metagenome library at a level of Log 4.11 CFU per g dry soil.

scholarly journals Transduction, restriction and recombination patterns in Escherichia coli.

Genetics ◽  
1995 ◽  
Vol 139 (1) ◽  
pp. 35-43
Author(s):  
M McKane ◽  
R Milkman
Keyword(s):  
Escherichia Coli ◽  
Coli Strain ◽  
Chromosomal Dna ◽  
Bacteriophage P1 ◽  
E Coli ◽  
Sequence Patterns ◽  
Single Marker ◽  
Different Strains ◽  
Restriction Modification ◽  
Restriction Modification Systems

Abstract Chromosomal DNA from several Escherichia coli reference (ECOR) strains was transduced by bacteriophage P1 into E. coli strain K12 W3110 trpA33. Recombination patterns of the transductants were determined by restriction fragment length polymorphism over a 40-kb region centering on a single marker (trpA+) in the tryptophan operon. These experiments demonstrate that transduction between different strains of E. coli can result in recombinational replacements that are small in comparison to the entrant molecule (replacements average 8-14 kb, whereas P1 packages approximately 100 kb) often in a series of discrete segments. The transduction patterns generated resemble the natural mosaic sequence patterns of the ECOR strains described in previous work. Extensive polymorphisms in the restriction-modification systems of the ECOR strains are a possible explanation for the sequence patterns in nature. To test this possibility two transductants were back-transduced into strain K12 W3110 trpA33. The resulting patterns were strikingly different from the original transductions. The size of the replacements was greater, and no multiple replacements were observed, suggesting a role for restriction-modification systems in the transduction patterns and perhaps for the mosaic sequence patterns in nature.

scholarly journals Differential molecular approach and ESBL detection from Klebsiella pneumoniae and Escherichia coli isolated from the supraglottic region of patients undergoing mechanical ventilation in an intensive care unit

2018 ◽  
Vol 66 (4) ◽  
pp. 581-587
Author(s):  
Olga Lucía Tovar ◽  
Gloria Inés Estrada ◽  
María Cristina Florián ◽  
Alejandro Uribe ◽  
Carlos Andrés Marulanda ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Intensive Care Unit ◽  
Mechanical Ventilation ◽  
Intensive Care ◽  
Klebsiella Pneumoniae ◽  
Molecular Characterization ◽  
Coli Strain ◽  
E Coli ◽  
Infectious Origin ◽  
Inert Surfaces

Introduction: Given their ability for colonizing the supraglottic region, desiccation tolerance, resistance to β-lactam antibiotics, and adherence to both inert surfaces and epithelial cells, Klebsiella pneumoniae and Escherichia coli are potentially pathogenic microorganisms for patients undergoing mechanical ventilation in an intensive care unit (ICU).Objective: To perform a molecular characterization and detection of extended spectrum β-lactamases (ESBL) in K. pneumoniae and E. coli strains isolated from the supraglottic region of patients undergoing mechanical ventilation in an ICU.Materials and methods: A descriptive study was conducted in 18 isolates. Disk diffusion technique was used for detecting ESBL-producing bacteria. Molecular characterization was made by BOX-PCR technique, while ESBL production was confirmed by testing the isolates against cefotaxime and ceftazidime, alone and in combination with clavulanic acid.Results: a K. pneumoniae strain and another E. coli strain were confirmed as ESBL producers. A divergence greater than 50% was observed in most of the strains; besides non-infectious origin strains resistant to third generation cephalosporins were found.Conclusion: The polyclonality found in this study might indicate that most of the strains belong to each patient’s microbiota.

Transport of escherichia coli through soil to groundwater traced by randomly amplified polymorphic DNA (RAPD)

1997 ◽  
Vol 35 (11-12) ◽  
pp. 351-357 ◽  
Author(s):  
R. Rothmaier ◽  
A. Weidenmann ◽  
K. Botzenhart
Keyword(s):  
Negative Impact ◽  
Random Amplified Polymorphic Dna ◽  
Coli Strain ◽  
Soil Samples ◽  
Test Field ◽  
Liquid Manure ◽  
E Coli ◽  
Rapd Pattern ◽  
Geological Situation ◽  
Different Strains

Isolates (50) of E. coli obtained from liquid manure (20 bovine, 20 porcine) were genotyped using random amplified polymorphic DNA (RAPD). Typing revealed 9 and 14 different strains in bovine and porcine liquid manure respectively with no strains in common. One porcine strain, showing a simple RAPD pattern, was subcultured and spread on a test field (1.5l/m2 at 1010 cfu/l) in a drinking water protection zone with loamy to sandy sediments in the Donauried area, Baden-Wurttemberg. Soil samples and groundwaters were collected at monthly intervals October 1994 – June 1995 during which 114 E. coli isolates were recovered. The first occurrence and maximum concentration of E. coli in soil samples taken from more than 20cm depth was in January 1995, declining rapidly with depth and time. All isolates from soil and only one from groundwater showed the RAPD pattern of the spread E. coli strain. The results could not demonstrate a severe negative impact of the spreading of liquid manure on the bacteriological quality of the groundwater in the given geological situation. The distinct strain patterns found in different kinds of liquid manure suggest that genotyping of E. coli by RAPD may be an adequate tool for tracing sources of faecal contamination.

scholarly journals Regulation of Expression of the TIR-Containing Protein C Gene of the Uropathogenic Escherichia coli Strain CFT073

Pathogens ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 549
Author(s):  
Julia Ittensohn ◽  
Jacqueline Hemberger ◽  
Hannah Griffiths ◽  
Maren Keller ◽  
Simone Albrecht ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein C ◽  
Interleukin 1 ◽  
Coli Strain ◽  
Uropathogenic Escherichia Coli ◽  
Reporter Construct ◽  
Regulation Of Expression ◽  
E Coli ◽  
Strain Cft073 ◽  
Myeloid Differentiation Factor

The uropathogenic Escherichia coli strain CFT073 causes kidney abscesses in mice Toll/interleukin-1 receptor domain-containing protein C (TcpC) dependently and the corresponding gene is present in around 40% of E. coli isolates of pyelonephritis patients. It impairs the Toll-like receptor (TLR) signaling chain and the NACHT leucin-rich repeat PYD protein 3 inflammasome (NLRP3) by binding to TLR4 and myeloid differentiation factor 88 as well as to NLRP3 and caspase-1, respectively. Overexpression of the tcpC gene stopped replication of CFT073. Overexpression of several tcpC-truncation constructs revealed a transmembrane region, while its TIR domain induced filamentous bacteria. Based on these observations, we hypothesized that tcpC expression is presumably tightly controlled. We tested two putative promoters designated P1 and P2 located at 5′ of the gene c2397 and 5′ of the tcpC gene (c2398), respectively, which may form an operon. High pH and increasing glucose concentrations stimulated a P2 reporter construct that was considerably stronger than a P1 reporter construct, while increasing FeSO4 concentrations suppressed their activity. Human urine activated P2, demonstrating that tcpC might be induced in the urinary tract of infected patients. We conclude that P2, consisting of a 240 bp region 5′ of the tcpC gene, represents the major regulator of tcpC expression.

Polyester Cloth–Based Hybridization Array System for Identification of Enterohemorrhagic Escherichia coli Serogroups O26, O45, O103, O111, O121, O145, and O157

2012 ◽  
Vol 75 (9) ◽  
pp. 1691-1697 ◽  
Author(s):  
BURTON W. BLAIS ◽  
MARTINE GAUTHIER ◽  
MYLÈNE DESCHÊNES ◽  
GEORGE HUSZCZYNSKI
Keyword(s):  
Escherichia Coli ◽  
Marker Gene ◽  
Enterohemorrhagic Escherichia Coli ◽  
Oligonucleotide Probes ◽  
E Coli ◽  
Polyester Cloth ◽  
Gene Profiles ◽  
Pcr Products ◽  
Different Strains ◽  
Immunoenzymatic Assay

A cloth-based hybridization array system (CHAS) was developed for the identification of foodborne colony isolates of seven priority enterohemorrhagic Escherichia coli (EHEC-7) serogroups targeted by U.S. food inspection programs. Gene sequences associated with intimin; Shiga-like toxins 1 and 2; and the antigenic markers O26, O45, O103, O111, O121, O145, and O157 were amplified in a multiplex PCR incorporating a digoxigenin label, and detected by hybridization of the PCR products with an array of specific oligonucleotide probes immobilized on a polyester cloth support, with subsequent immunoenzymatic assay of the captured amplicons. The EHEC-7 CHAS exhibited 100% inclusivity and 100% exclusivity characteristics with respect to detection of the various markers among 89 different E. coli strains, with various marker gene profiles and 15 different strains of non–E. coli bacteria.

scholarly journals Quantifying within- and between-animal variation and uncertainty associated with counts of Escherichia coli O157 occurring in naturally infected cattle faeces

2008 ◽  
Vol 6 (31) ◽  
pp. 169-177 ◽  
Author(s):  
S.E Robinson ◽  
P.E Brown ◽  
E.J Wright ◽  
C.A Hart ◽  
N.P French
Keyword(s):  
Escherichia Coli ◽  
Statistical Model ◽  
Human Infection ◽  
Microbial Count ◽  
Infected Cattle ◽  
Explanatory Variables ◽  
E Coli ◽  
Fixed And Random Effects ◽  
Cattle Faeces ◽  
Multiple Samples

Cattle faeces are considered the most important reservoir for human infection with Escherichia coli O157. We have previously described shedding of E. coli O157 in the faeces of naturally infected cattle cohorts. However, the data require further investigation to quantify the uncertainty and variability in the estimates previously presented. This paper proposes a method for analysing both the presence and the quantity of E. coli O157 in cattle faecal samples, using two isolation procedures, one of which enumerates E. coli O157. The combination of these two measurements, which are fundamentally different in nature and yet measuring a common outcome, has necessitated the development of a novel statistical model for ascertaining the contribution of the various components of variation (both natural and observation induced) and for judging the influence of explanatory variables. Most of the variation within the sampling hierarchy was attributable to multiple samples from the same animal. The contribution of laboratory-level variation was found to be low. After adjusting for fixed and random effects, short periods of increased intensity of shedding were identified in individual animals. We conclude that within-animal variation is greater than between animals over time, and studies aiming to elucidate the dynamics of shedding should focus resources, sampling more within than between animals. These findings have implications for the identification of persistent high shedders and for assessing their role in the epidemiology of E. coli O157 in cattle populations. The development of this non-standard statistical model may have many applications to other microbial count data.

scholarly journals Transcriptomic Analysis of Shiga Toxin-Producing Escherichia coli during Initial Contact with Cattle Colonic Explants

2020 ◽  
Vol 8 (11) ◽  
pp. 1662
Author(s):  
Zachary R. Stromberg ◽  
Rick E. Masonbrink ◽  
Melha Mellata
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Bacterial Colonization ◽  
Coli Strain ◽  
Fold Change ◽  
Initial Contact ◽  
Lon Protease ◽  
E Coli ◽  
Log2 Fold Change ◽  
Colonization Factors

Foodborne pathogens are a public health threat globally. Shiga toxin-producing Escherichia coli (STEC), particularly O26, O111, and O157 STEC, are often associated with foodborne illness in humans. To create effective preharvest interventions, it is critical to understand which factors STEC strains use to colonize the gastrointestinal tract of cattle, which serves as the reservoir for these pathogens. Several colonization factors are known, but little is understood about initial STEC colonization factors. Our objective was to identify these factors via contrasting gene expression between nonpathogenic E. coli and STEC. Colonic explants were inoculated with nonpathogenic E. coli strain MG1655 or STEC strains (O26, O111, or O157), bacterial colonization levels were determined, and RNA was isolated and sequenced. STEC strains adhered to colonic explants at numerically but not significantly higher levels compared to MG1655. After incubation with colonic explants, flagellin (fliC) was upregulated (log2 fold-change = 4.0, p < 0.0001) in O157 STEC, and collectively, Lon protease (lon) was upregulated (log2 fold-change = 3.6, p = 0.0009) in STEC strains compared to MG1655. These results demonstrate that H7 flagellum and Lon protease may play roles in early colonization and could be potential targets to reduce colonization in cattle.

scholarly journals Efficient production of (S)-1-phenyl-1, 2-ethanediol using xylan as co-substrate by a coupled multi-enzyme Escherichia coli system

2020 ◽  
Author(s):  
Junchao Rao ◽  
Rongzhen Zhang ◽  
Guanyu Xu ◽  
Lihong Li ◽  
Yan Xu
Keyword(s):  
Escherichia Coli ◽  
Glucose Dehydrogenase ◽  
Optical Purity ◽  
Carbonyl Reductase ◽  
Cofactor Regeneration ◽  
Efficient Production ◽  
Chiral Synthesis ◽  
E Coli ◽  
Concomitant Reduction ◽  
Different Strains

Abstract Background: ( S )-1-phenyl-1,2-ethanediol is an important chiral intermediate in the synthesis of liquid crystals and chiral biphosphines.(S)-carbonyl reductase II from Candida parapsilosis catalyzes the conversion of 2-hydroxyacetophenone to ( S )-1-phenyl-1,2-ethanediol with NADPH as a cofactor. Glucose dehydrogenase with a Ala258Phe mutation is able to catalyze the oxidation of xylose with concomitant reduction of NADP + to NADPH, while endo-β-1,4-xylanase 2 catalyzes the conversion of xylan to xylose. In the present work, the Ala258Phe glucose dehydrogenase mutant and endo-β-1,4-xylanase 2 were introduced into the ( S )-carbonyl reductase II-mediated chiral pathway to strengthen cofactor regeneration by using xylan as a naturally abundant co-substrate. Results: We constructed several coupled multi-enzyme systems by introducing ( S )-carbonyl reductase II, the A258F glucose dehydrogenase mutant and endo-β-1,4-xylanase 2 into Escherichia coli . Different strains were produced by altering the location of the encoding genes on the plasmid. Only recombinant E. coli /pET-G-S-2 expressed all three enzymes, and this strain produced ( S )-1-phenyl-1,2-ethanediol from 2-hydroxyacetophenone as a substrate and xylan as a co-substrate. The optical purity was 100% and the yield was 98.3% (6 g/L 2-HAP) under optimal conditions of 35°C, pH 6.5 and a 2:1 substrate-co-substrate ratio. The introduction of A258F glucose dehydrogenase and endo-β-1,4-xylanase 2 into the ( S )-carbonyl reductase II-mediated chiral pathway caused a 54.6% increase in yield, and simultaneously reduced the reaction time from 48 h to 28 h. Conclusions: This study demonstrates efficient chiral synthesis using a pentose as a co-substrate to enhance cofactor regeneration. This provides a new approach for enantiomeric catalysis through the inclusion of naturally abundant materials.

scholarly journals Removal of Methylene blue, Escherichia coli and Pseudomonus aeruginosa by Adsorption Process of Activated Carbon Produced from Moringa oleifera Bark

2021 ◽  
Author(s):  
Md. Shahin Azad ◽  
Syaza Azhari ◽  
Mohd Sukri Hassan
Keyword(s):  
Escherichia Coli ◽  
Methylene Blue ◽  
Activated Carbon ◽  
Moringa Oleifera ◽  
Adsorption Process ◽  
Diffusion Mechanism ◽  
Bacterial Load ◽  
Batch Adsorption ◽  
Maximum Adsorption Capacity ◽  
E Coli

The utilization of biopolymer derived from Moringa oleifera bark using ZnCl2 and H2SO4 as activating agents for eliminating Methylene blue, Escherichia coli and Pseudomonas aeruginosa from producing wastewater. In this study, Methylene blue and both bacteria were effectively adsorbed by activated carbon with lowest dosage. The activated carbon was prepared from natural-by product of Moringa oleifera bark by pyrolysis in a furnace at 700°C for 1 h. The characteristics of activated carbon have been determined using Scanning Electron Microscopy (SEM), Brunauer-Emmett-Teller (BET), pHzpc (zero point charge), and FTIR spectroscopy. The obtained result were closely fitted with Freundlich isotherm model and adsorption kinetics follow the pseudo-second order model with the highest value of correlation coefficient (R2~1). Adsorption quantity was dose dependent and bacteria were maximum adsorbed using 10 mg of activated carbon as well as 25mg for methylene blue. The maximum adsorption capacity showed within 1 hour. The bacterial load was reduced by 98% for E. coli, 96% for P. aeruginosa as well as methylene blue reduced 94.2% from aqueous solution using batch adsorption methods. Adsorption process controlled by film diffusion mechanism. These result proposed that the activated carbon of Moringa oleifera can be used as a good adsorbent for the removal of Methylene blue, E. coli and P. aeruginosa.

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