scholarly journals An Engineered R-Type Pyocin Is a Highly Specific and Sensitive Bactericidal Agent for the Food-Borne Pathogen Escherichia coli O157:H7

2009 ◽  
Vol 53 (7) ◽  
pp. 3074-3080 ◽  
Author(s):  
Dean Scholl ◽  
Mike Cooley ◽  
Steve R. Williams ◽  
Dana Gebhart ◽  
David Martin ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Agents ◽  
Cell Envelope ◽  
Molecular Techniques ◽  
Spike Protein ◽  
Tail Fiber ◽  
Primary Spectrum ◽  
E Coli ◽  
The Family ◽  
Tail Spike

ABSTRACT Some strains of Pseudomonas aeruginosa produce R-type pyocins, which are high-molecular-weight phage tail-like protein complexes that have bactericidal activity against other Pseudomonas strains. These particles recognize and bind to bacterial surface structures via tail fibers, their primary spectrum determinant. R-type pyocins kill the cell by contracting a sheath-like structure and inserting their hollow core through the cell envelope, resulting in dissipation of the cellular membrane potential. We have retargeted an R-type pyocin to Escherichia coli O157:H7 by fusing a tail spike protein from an O157-specific phage, φV10, to the pyocin tail fiber. The φV10 tail spike protein recognizes and degrades the O157 lipopolysaccharide. This engineered pyocin, termed AVR2-V10, is sensitive and specific, killing 100% of diverse E. coli O157:H7 isolates but no other serotypes tested. AVR2-V10 can kill E. coli O157:H7 on beef surfaces, making it a candidate agent for the elimination of this pathogen from food products. All rare AVR2-V10-resistant mutants isolated and examined have lost the ability to produce the O157 antigen and are expected to have compromised virulence. In addition, E. coli O157:H7 exposed to and killed by AVR2-V10 do not release Shiga toxin, as is often the case with many antibiotics, suggesting potential therapeutic applications. The demonstration that a novel R-type pyocin can be created in the laboratory by fusing a catalytic tail spike from the family Podoviridae to a tail fiber of a member of the family Myoviridae is evidence that the plasticity observed among bacteriophage tail genes can, with modern molecular techniques, be exploited to produce nonnatural, targeted antimicrobial agents.

scholarly journals Investigation of Pathogenic Escherichia Coli from Diarrheic Calves in Selective Area of Bangladesh

2015 ◽  
Vol 13 (1) ◽  
pp. 45-51 ◽  
Author(s):  
AKMA Islam ◽  
M Rahman ◽  
A Nahar ◽  
A Khair ◽  
MM Alam
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Agents ◽  
Biochemical Properties ◽  
Molecular Techniques ◽  
Diffusion Method ◽  
Molecular Technique ◽  
Isolation And Identification ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Diarrheic Calves

Molecular technique was used to investigate the prevalence of virulent diarrheic genes in pathogenic Escherichia coli and their antibiotic sensitivity patterns. A hundred samples from 100 different diarrheic calves from mid-north-western part of Bangladesh were screened for the presence of virulence factors associated with diarrhea. Following isolation and identification on the basis of cultural, morphological and biochemical properties, the presence of the virulence genes such as eaeA, bfpA, elt, est, stx1 and stx2 were examined using PCR. Antimicrobial susceptibility of 57 E. coli was determined by agar disk diffusion method for 8 antimicrobial agents. Out of 100 samples 57 (57%) were found to be positive for E. coli and their distribution rates according to their age, breed and sex were  66.7% ( 6 days old ), 85.7% (Sahiwal breed) and in  64.2 % (female calves) respectively. Among 57 E. coli isolates, only 16 isolates were analyzed for the detection of the said genes. Among them, only eaeA gene was detected in 2 E. coli isolates (12.5 %). Antibiotic resistance patterns revealed that Oxacillin, Rifampicin and Penicillin were  100% resistant followed by Erythromycin which was more than 80% resistant. In case of Amoxicillin and Tetracycline, about 59.65% and 61.40% were found to be resistant respectively whereas all 57 E. coli isolates showed moderately susceptible (30%) to Cefuroxime, a second generation Cephalosporin. Therefore, none of the eight antimicrobials studied can not be recommended as single best therapeutic agent for the treatment of neonatal calf diarrhea. In addition, this study indicated that diarrhea in calves in these locations can be ascribed to mainly Enteropathogenic E. coli (EPEC) which was atypical (only contained the eaeA genes but not bfpA). However, further studies are necessary to characterize the isolated eaeA gene positive E. coli by serotyping, tissue culture assay and other molecular techniques to find out the potentiality of those virulent genes contributing pathogenicity of E. coli causing diarrhea in calves.DOI: http://dx.doi.org/10.3329/bjvm.v13i1.23716Bangl. J. Vet. Med. (2015). 13 (1): 45-51

scholarly journals Phenotypic Detection of Extended-Spectrum ß-lactamase Produced by Escherichia coli Isolates from Health facilities in Otukpo, Benue State Nigeria

2021 ◽  
Vol 2 (2) ◽  
pp. 7-13
Author(s):  
PS Utulo ◽  
EU Umeh ◽  
GM Gberikon ◽  
PO Abba
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Agents ◽  
Health Facilities ◽  
Molecular Techniques ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Esbl Production ◽  
E Coli ◽  
Extended Spectrum ◽  
Microbiological Methods

Beta-lactamase production by Escherichia coli and other bacteria is one of the most important mechanism of resistance to beta-lactam antibiotics such as Penicillins and Cephalosporins which are the most commonly used antibiotics in the treatment of bacterial infections in Hospitals. Resistance to antibiotics is emerging worldwide as a threat to human health resulting in limitations of therapeutic options. Susceptibility tests of microorganisms to antimicrobial agents do not reveal production of Extended-Spectrum ß-lactamase, hence the need to detect their presence by phenotypic or molecular techniques. This study is aimed at determining the prevalence of ESBL-production by Escherichia coli isolates from clinical samples collected from some selected health facilities in Otukpo, using phenotypic detection. Four hundred (400) specimens were collected from four different health facilities in Otukpo. Specimens collected included urine, blood, stool and wound swabs from 222 females and 178 males. Standard microbiological methods were employed for isolation, identification and characterization of Escherichia coli isolates from these specimens. Antibiotic susceptibility of the E. coli isolates was determined using Kirby-Bauer disc diffusion method while double disc synergy test (DDST) was used for ESBL production. Sixty-eight, (17%) of the 400 specimens tested for ESBL production were positive. Isolates from stool 22(22.7%) had the highest prevalence of ESBL-producing E. coli, followed by isolates from blood 4(21.4%). Urine isolates had the least prevalence 31(13.8%). Isolates from female subjects, 38(17.1%) had higher positivity rate than their male counterpart 30(16.9%). Otukpo General Hospital had the highest prevalence 20(20.0%) of ESBL followed by Otia Hospital 18(18.0%). Comprehensive health center, Otukpo had the least prevalence 14(14.0%). There is a high prevalence of ESBL production by Escherichia coli isolated from health facilities in Otukpo. Routine drug resistance surveillance therefore has become necessary to guide the appropriate and judicious antibiotic use.

Antimicrobial Effects of Zataria multiflora and Ocimum basilicum on Escherichia coli O157:H7 During Ripening of Traditional Lighvan Cheese

2020 ◽  
Vol 16 (3) ◽  
pp. 373-380
Author(s):  
Mohammad B. Zendeh ◽  
Vadood Razavilar ◽  
Hamid Mirzaei ◽  
Khosrow Mohammadi
Keyword(s):  
Escherichia Coli ◽  
Essential Oils ◽  
Dairy Products ◽  
Antimicrobial Agents ◽  
Escherichia Coli O157 ◽  
Zataria Multiflora ◽  
Antimicrobial Effects ◽  
E Coli ◽  
Common Causes ◽  
Lighvan Cheese

Background: Escherichia coli O157:H7 is one of the most common causes of contamination in Lighvan cheese processing. Using from natural antimicrobial essential oils is applied method to decrease the rate of microbial contamination of dairy products. The present investigation was done to study the antimicrobial effects of Z. multiflora and O. basilicum essential oils on survival of E. coli O157:H7 during ripening of traditional Lighvan cheese. Methods: Leaves of the Z. multiflora and O. basilicum plants were subjected to the Clevenger apparatus. Concentrations of 0, 100 and 200 ppm of the Z. multiflora and 0, 50 and 100 ppm of O. basilicum essential oils and also 103 and 105 cfu/ml numbers of E. coli O157:H7 were used. The numbers of the E. coli O157:H7 bacteria were analyzed during the days 0, 30, 60 and 90 of the ripening period. Results: Z. multiflora and O. basilicum essential oils had considerable antimicrobial effects against E. coli O157:H7. Using the essential oils caused decrease in the numbers of E. coli O157:H7 bacteria in 90th days of ripening (P <0.05). Using from Z. multiflora at concentration of 200 ppm can reduce the survival of E. coli O157:H7 in Lighvan cheese. Conclusion: Using Z. multiflora and O. basilicum essential oils as good antimicrobial agents can reduce the risk of foodborne bacteria and especially E. coli O157:H7 in food products.

scholarly journals Impact of Feed Supplementation with Antimicrobial Agents on Growth Performance of Broiler Chickens, Clostridium perfringens and Enterococcus Counts, and Antibiotic Resistance Phenotypes and Distribution of Antimicrobial Resistance Determinants in Escherichia coli Isolates

2007 ◽  
Vol 73 (20) ◽  
pp. 6566-6576 ◽  
Author(s):  
Moussa S. Diarra ◽  
Fred G. Silversides ◽  
Fatoumata Diarrassouba ◽  
Jane Pritchard ◽  
Luke Masson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Broiler Chickens ◽  
Antimicrobial Agents ◽  
Growth Promoters ◽  
Feed Supplementation ◽  
E Coli ◽  
Resistance Determinants ◽  
Class 1

ABSTRACT The effects of feed supplementation with the approved antimicrobial agents bambermycin, penicillin, salinomycin, and bacitracin or a combination of salinomycin plus bacitracin were evaluated for the incidence and distribution of antibiotic resistance in 197 commensal Escherichia coli isolates from broiler chickens over 35 days. All isolates showed some degree of multiple antibiotic resistance. Resistance to tetracycline (68.5%), amoxicillin (61.4%), ceftiofur (51.3%), spectinomycin (47.2%), and sulfonamides (42%) was most frequent. The levels of resistance to streptomycin, chloramphenicol, and gentamicin were 33.5, 35.5, and 25.3%, respectively. The overall resistance levels decreased from day 7 to day 35 (P < 0.001). Comparing treatments, the levels of resistance to ceftiofur, spectinomycin, and gentamicin (except for resistance to bacitracin treatment) were significantly higher in isolates from chickens receiving feed supplemented with salinomycin than from the other feeds (P < 0.001). Using a DNA microarray analysis capable of detecting commonly found antimicrobial resistance genes, we characterized 104 tetracycline-resistant E. coli isolates from 7- to 28-day-old chickens fed different growth promoters. Results showed a decrease in the incidence of isolates harboring tet(B), bla TEM, sulI, and aadA and class 1 integron from days 7 to 35 (P < 0.01). Of the 84 tetracycline-ceftiofur-resistant E. coli isolates, 76 (90.5%) were positive for bla CMY-2. The proportions of isolates positive for sulI, aadA, and integron class 1 were significantly higher in salinomycin-treated chickens than in the control or other treatment groups (P < 0.05). These data demonstrate that multiantibiotic-resistant E. coli isolates can be found in broiler chickens regardless of the antimicrobial growth promoters used. However, the phenotype and the distribution of resistance determinants in E. coli can be modulated by feed supplementation with some of the antimicrobial agents used in broiler chicken production.

Interaction of Gram-Negative Bacteria with the Lysosomal Fraction of Polymorphonuclear Leukocytes II. Changes in the Cell Envelope of Escherichia coli

1970 ◽  
Vol 1 (3) ◽  
pp. 311-318
Author(s):  
D. Friedberg ◽  
I. Friedberg ◽  
M. Shilo
Keyword(s):  
Escherichia Coli ◽  
Polymorphonuclear Leukocytes ◽  
Cytoplasmic Membrane ◽  
Morphological Changes ◽  
Cell Envelope ◽  
Gram Negative Bacteria ◽  
Intact Cells ◽  
Lysosomal Fraction ◽  
E Coli ◽  
Absorbing Material

Interaction of lysosomal fraction with Escherichia coli caused damage to the cell envelope of these intact cells and to the cytoplasmic membrane of E. coli spheroplasts. The damage to the cytoplasmic membrane was manifested in the release of 260-nm absorbing material and β-galactosidase from the spheroplasts, and by increased permeability of cryptic cells to O -nitrophenyl-β- d -galactopyranoside; damage to the cell wall was measured by release of alkaline phosphatase. Microscope observation showed morphological changes in the cell envelope.

scholarly journals Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

2007 ◽  
Vol 59 (2) ◽  
pp. 508-512 ◽  
Author(s):  
B.R. Paneto ◽  
R.P. Schocken-Iturrino ◽  
C. Macedo ◽  
E. Santo ◽  
J.M. Marin
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Raw Milk ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Polymerase Chain ◽  
Raw Milk Cheese

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).

scholarly journals Genotyping and antimicrobial resistance in Escherichia coli from pig carcasses

2017 ◽  
Vol 37 (11) ◽  
pp. 1253-1260 ◽  
Author(s):  
Caroline Pissetti ◽  
Gabriela Orosco Werlang ◽  
Jalusa Deon Kich ◽  
Marisa Cardoso
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Antimicrobial Agents ◽  
Commensal Bacteria ◽  
Pfge Analysis ◽  
Gene Detection ◽  
E Coli ◽  
Antimicrobial Resistance Profile ◽  
Resistant Strains ◽  
Pig Carcasses

ABSTRACT: The increasing antimicrobial resistance observed worldwide in bacteria isolated from human and animals is a matter of extreme concern and has led to the monitoring of antimicrobial resistance in pathogenic and commensal bacteria. The aim of this study was to evaluate the antimicrobial resistance profile of Escherichia coli isolated from pig carcasses and to assess the occurrence of relevant resistance genes. A total of 319 E. coli isolates were tested for antimicrobial susceptibility against different antimicrobial agents. Moreover, the presence of extended-spectrum β-lactamase (ESBL) and inducible ampC-β-lactamase producers was investigated. Eighteen multi-resistant strains were chosen for resistance gene detection and PFGE characterization. The study showed that resistance to antimicrobials is widespread in E. coli isolated from pig carcasses, since 86.2% of the strains were resistant to at least one antimicrobial and 71.5% displayed multi-resistance profiles. No ampC-producing isolates were detected and only one ESBL-producing E. coli was identified. Genes strA (n=15), floR (n=14), aac(3)IVa (n=13), tetB (n=13), sul2 (n=12), tetA (n=11), aph(3)Ia (n=8) and sul3 (n=5) were detected by PCR. PFGE analysis of these multi-resistant E. coli strains showed less than 80% similarity among them. We conclude that antimicrobial multi-resistant E. coli strains are common on pig carcasses and present highly diverse genotypes and resistance phenotypes and genotypes.

scholarly journals Secondary transmissions during the outbreak of Shiga toxin-producing Escherichia coli O104 in Hesse, Germany, 2011

2011 ◽  
Vol 16 (31) ◽  
Author(s):  
A M Hauri ◽  
U Götsch ◽  
I Strotmann ◽  
J Krahn ◽  
G Bettge-Weller ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Case Definition ◽  
Case Definitions ◽  
Secondary Transmission ◽  
E Coli ◽  
Outbreak Area ◽  
The Family ◽  
Satellite Clusters ◽  
Outbreak Case

During the recent outbreak of Shiga toxin-producing Escherichia coli (STEC) O104:H4 in Germany most cases notified in the State of Hesse (6 million inhabitants) were linked to satellite clusters or had travelled to the outbreak area in northern Germany. Intensified surveillance was introduced to rapidly identify cases not linked to known clusters or cases and thus to obtain timely information on possible further contaminated vehicles distributed in Hesse, as well to describe the risk of secondary transmission among known cases. As of 2 August 2011*, 56 cases of haemolytic uraemic syndrome (HUS) including two fatal cases, and 124 cases of STEC gastroenteritis meeting the national case definitions have been reported in Hesse. Among the 55 HUS and 81 STEC gastroenteritis cases that met the outbreak case definition, one HUS case and eight STEC gastroenteritis cases may have acquired their infection through secondary transmission. They include six possible transmissions within the family, two possible nosocomial and one possible laboratory transmission. Our results do not suggest an increased transmissibility of the outbreak strain compared to what is already known about E. coli O157 and other STEC serotypes.

scholarly journals Molecular characterisation of antimicrobial resistance and virulence genes in Escherichia coli strains isolated from diarrhoeic and healthy rabbits in Tunisia

2020 ◽  
Vol 28 (2) ◽  
pp. 81
Author(s):  
Raouia Ben Rhouma ◽  
Ahlem Jouini ◽  
Amira Klibi ◽  
Safa Hamrouni ◽  
Aziza Boubaker ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Virulence Genes ◽  
Antimicrobial Agents ◽  
Antibiotic Resistance Genes ◽  
Virulence Gene ◽  
Diffusion Method ◽  
Class 1 Integron ◽  
E Coli ◽  
Class 1

The purpose of this study was to identify <em>Escherichia coli</em> isolates in diarrhoeic and healthy rabbits in Tunisia and characterise their virulence and antibiotic resistance genes. In the 2014-2015 period, 60 faecal samples from diarrhoeic and healthy rabbits were collected from different breeding farms in Tunisia. Susceptibility to 14 antimicrobial agents was tested by disc diffusion method and the mechanisms of gene resistance were evaluated using polymerase chain reaction and sequencing methods. Forty <em>E. coli</em> isolates were recovered in selective media. High frequency of resistance to tetracycline (95%) was detected, followed by different levels of resistance to sulphonamide (72.5%), streptomycin (62.5%), trimethoprim-sulfamethoxazole (60%), nalidixic acid (32.5%), ampicillin (37.5%) and ticarcillin (35%). <em>E. coli</em> strains were susceptible to cefotaxime, ceftazidime and imipenem. Different variants of bla<sub>TEM</sub>, <em>tet</em>, <em>sul</em> genes were detected in most of the strains resistant to ampicillin, tetracycline and sulphonamide, respectively. The presence of class 1 integron was studied in 29 sulphonamide-resistant <em>E. coli</em> strains from which 15 harboured class 1 integron with four different arrangements of gene cassettes, <em>dfrA17</em>+<em>aadA5</em> (n=9), <em>dfrA1</em> + <em>aadA1</em> (n=4), <em>dfrA12</em> + <em>addA2</em> (n=1), <em>dfrA12</em>+<em>orf</em>+<em>addA2</em> (n=1). The <em>qnrB</em> gene was detected in six strains out of 13 quinolone-resistant <em>E. coli</em> strains. Seventeen <em>E. coli</em> isolates from diarrhoeic rabbits harboured the enteropathogenic eae genes associated with different virulence genes tested (<em>fimA</em>, <em>cnf1</em>, <em>aer</em>), and affiliated to B2 (n=8) and D (n=9) phylogroups. Isolated <em>E. coli</em> strains from healthy rabbit were harbouring <em>fim A</em> and/or <em>cnf1</em> genes and affiliated to A and B1 phylogroups. This study showed that <em>E. coli</em> strains from the intestinal tract of rabbits are resistant to the widely prescribed antibiotics in medicine. Therefore, they constitute a reservoir of antimicrobial-resistant genes, which may play a significant role in the spread of antimicrobial resistance. In addition, the eae virulence gene seemed to be implicated in diarrhoea in breeder rabbits in Tunisia.

In vitro activity of antimicrobial agents with and without sulbactam, EDTA and sulbactam-EDTA on ESBLs-producing Escherichia coli isolated from healthy Tibetan yaks

2020 ◽  
Author(s):  
Baoguang Liu ◽  
Xiaoling Yuan ◽  
Yiheng Chen ◽  
Xiaoshen Li ◽  
Ming Bai ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Nasal Swab ◽  
Antimicrobial Agents ◽  
Synergistic Effects ◽  
E Coli ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Third Generation Cephalosporins ◽  
Enhance Activity

Abstract Background The spread of ESBLs-producing bacteria has been strikingly rapid in many regions of the world and it causes therapeutic difficulties in everyday practice. The aims of this study were to investigate the prevalence and susceptibilities of ESBLs-producing Escherichia coli isolates from healthy Tibetan yaks in China, to evaluate the activity of drug combinations on ESBLs-producing E. coli isolates. Methods From July 2018 to August 2019, a total of 750 nasal swab samples were tested for the presence of E. coli and ESBLs-producing strains. The MICs of 11 antimicrobial agents alone and combinations with sulbactam, EDTA or sulbactam-EDTA against 240 ESBLs-producing E.coli strains were determined by the broth microdilution method. Results Overall, 59.87% (n = 449) of the samples were positive for E. coli, 240 (53.45%) of 449 E. coli isolates were confirmed to be ESBLs-producing. The addition of sulbactam to the third generation cephalosporins, amikacin and fosfomycin for all isolates resulted in low MICs, increasing the level of susceptibility from 0, 0 and 0% to 50 ~ 87.5, 4.2 and 100% respectively. The addition of EDTA to fluoroquinolones, doxycycline, florfenicol, amikacin and fosfomycin, showed improved activities and resulted in low MICs, increasing the level of susceptibility from 0, 0, 8.3, 0 and 0% to 4.2 ~ 29.2, 33.3, 33.3, 66.7 and 45.8%, respectively. All other antibacterials (except fluoroquinolones, doxycycline and florfenicol), when combined with sulbactam-EDTA, were found to be more active than combinations only with sulbactam or with EDTA against most of isolates, with lower MIC50s and MIC90s. Conclusion In conclusion, ESBLs-producing E. coli isolates were widespread in healthy Tibetan yaks in China. ESBLs-producing E. coli isolates exhibited varying degrees of multidrug resistance. This study these findings suggested that sulbactam can enhance activity of β-lactams and some non-β-lactams of antimicrobial agents and had a synergistic effects with EDTA in improving activities of some families of antimicrobials.

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