scholarly journals Isolation and characterization of Escherichia coli phage and enhance its bactericidal activity that collaborative with kanamycin sulfate

2020 ◽  
Author(s):  
Liming Jiang ◽  
Rui Zheng

Abstract Background: Escherichia coli is the most important and widespread bacteria in worldwide, which mainly found in contaminated food, human and animal faeces. Unfortunately, Some of E. coli strains are multidrug-resistant (MDR) pathogen leading significant public health concern globally. Biofilm is a multicellular community of microorganisms. Phages and their derivatives are ideal candidates for replacing or compensating for antibiotic problems in the future. Method: Here, we aimed to isolation and characterization of Escherichia coli phage and research its bactericidal activity that individually or collaborative with kanamycin sulfateResults: In this study, three virulent phages Flora, T4 and WJ were isolated from the laboratory and drug sample in Wuxi, China. It’s belonged to the Myoviridae family and optimum temperature is 42 ℃, optimum pH= 7, optimum MOI is 0.0001 and the genome size of Flora, T4 and WJ were 168, 909, 168903 and 168, 900 bp respectively. Flora has two exonuclease, whereas T4 and WJ have only one. Antibiotics have better bactericidal activity than phages in a low concentration medium of bacteria, nonetheless, phages have better bactericidal activity than antibiotics in a high concentration of bacteria, and that, collaboration of phages and antibiotics have better bactericidal activity effect than alone of phages or antibiotics in a low concentration medium of bacteria. Conclusion: The excellent performance of phage Flora for its therapeutic potential on clinic. The data of this study provided the strong evidence that the application of phage could reduce the growth and biofilm of E. coli that are important to maintain public health. Keywords: Escherichia coli, phage, lytic spectrum, biofilm, antibiotic

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Pouya Reshadi ◽  
Fatemeh Heydari ◽  
Reza Ghanbarpour ◽  
Mahboube Bagheri ◽  
Maziar Jajarmi ◽  
...  

Abstract Background Transmission of antimicrobial resistant and virulent Escherichia coli (E. coli) from animal to human has been considered as a public health concern. This study aimed to determine the phylogenetic background and prevalence of diarrheagenic E. coli and antimicrobial resistance in healthy riding-horses in Iran. In this research, the genes related to six main pathotypes of E. coli were screened. Also, genotypic and phenotypic antimicrobial resistance against commonly used antibiotics were studied, then phylo-grouping was performed on all the isolates. Results Out of 65 analyzed isolates, 29.23 % (n = 19) were determined as STEC and 6.15 % (n = 4) as potential EPEC. The most prevalent antimicrobial resistance phenotypes were against amoxicillin/clavulanic acid (46.2 %) and ceftriaxone (38.5 %). blaTEM was the most detected resistance gene (98.4 %) among the isolates and 26.15 % of the E. coli isolates were determined as multi-drug resistant (MDR). Three phylo-types including B1 (76.92 %), A (13.85 %) and D (3.08 %) were detected among the isolates. Conclusions Due to the close interaction of horses and humans, these findings would place emphasis on the pathogenic and zoonotic potential of the equine strains and may help to design antimicrobial resistance stewardship programs to control the dissemination of virulent and multi-drug resistant E. coli strains in the community.


1970 ◽  
Vol 18 ◽  
pp. 99-103 ◽  
Author(s):  
S Biswas ◽  
MAK Parvez ◽  
M Shafiquzzaman ◽  
S Nahar ◽  
MN Rahman

Context: Escherichia coli is shed in the feces of warm blooded animals and humans and thus potential for public health. Detection and characterization of E. coli in the ready-to-eat (RTE) foods concerns due to their presence indicates fecal contamination of the food.   Objective: To identify, characterize and RFLP pattern analysis of E. coli isolated from RTE foods vended in Islamic University campus, Kushtia.   Materials and Methods: Fifty samples from four types of consumed foods in six student halls of residence, some temporary restaurants of Islamic University, Kushtia were assessed for bacterial contamination by standard methods. Identification and characterization of E. coli isolates were performed using IMViC tests. Genomic DNA was used to perform RFLP pattern analysis.   Results: Thirty seven out of 50 (74%) examined samples of RTE foods had E. coli contamination. The highest number of E. coli was isolated from vegetable oriented RTE foods (90.90%) and fish, meat and cereals samples were also significantly E. coli positive. RFLP profiling of two E. coli isolates were observed.   Conclusion: The results of this study provide evidence that some RTE foods had unsatisfactory levels of contamination with E. coli. Thus street vended RTE food could be important potential vehicles for food-borne diseases. Molecular characterization may be exploited to identify food borne pathogen among different species.  Keywords: Ready-to-eat foods; Escherichia coli; RFLP pattern DOI: http://dx.doi.org/10.3329/jbs.v18i0.8783 JBS 2010; 18(0): 99-103


2019 ◽  
Author(s):  
Elizabeth Muligisa Muonga ◽  
Geoffrey Mainda ◽  
Mercy Mukuma ◽  
Geoffrey Kwenda ◽  
Bernard Hang'ombe ◽  
...  

Abstract Background Antimicrobial resistance (AMR) of foodborne pathogens is of public health concern, especially in developing countries like Zambia. This study was undertaken to determine the resistance profiles of Escherichia coli ( E. coli ) and Salmonella isolated from dressed broiler chickens purchased from open markets and supermarkets in Zambia.Results A total of 189 E. coli and five Salmonella isolates were isolated. Identification and confirmation of the isolates was done using Analytical Profile Index (API 20E) (Biomerieux ® ) and 16S rRNA sequencing. Antimicrobial susceptibility tests (AST) were performed using the Kirby Bauer disk diffusion technique using a panel of 10 different antibiotics and multiplex PCR was used to determine the presence of three target genes encoding for resistance: tetA, Sul1 and CTXM. AST results were entered and analyzed in WHONET 2018 software. A total of 189 E. coli and five Salmonella isolates were identified. Among the E. coli isolates, Tetracycline recorded the highest resistance of 79.4%, followed by Ampicillin 51.9%, Trimethoprim/Sulfamethoxazole 49.7%, Nalidixic Acid 24.3%, Chloramphenicol 16.4%, Cefotaxime 16.4%, Ciprofloxacin 10.1%, Colistin 7.4%, Amoxicillin/Clavulanic acid 6.9%, and Imipenem 1.1%. Two of the five Salmonella isolates were resistant to at least one antibiotic. Forty- seven (45.2%) of the isolates possessed at least one of the targeted resistance genes.Conclusion This study has demonstrated the presence of AMR E. coli and Salmonella on raw broiler chickens from both open markets and supermarkets. Such resistance is of public health concern and measures need to be put in place to regulate the use of these antimicrobials in poultry production.


1992 ◽  
Vol 283 (1) ◽  
pp. 87-90 ◽  
Author(s):  
C E Brunt ◽  
M C Cox ◽  
A G P Thurgood ◽  
G R Moore ◽  
G A Reid ◽  
...  

The cytochrome domain of flavocytochrome b2 (L-lactate dehydrogenase) was expressed in the bacterium Escherichia coli and a purification procedure was developed. When expressed in E. coli, the b2-cytochrome domain contains protohaem IX and has an electronic absorption spectrum identical with that of the cytochrome b2 ‘core’ produced by proteolytic cleavage of the enzyme isolated from yeast. The b2-cytochrome domain isolated from E. coli has an Mr of 10,500 and a redox potential of -31 +/- 2 mV. High-field n.m.r. studies indicate pKa values for the haem propionate groups to be 4.8 and 4.6, consistent with these groups being exposed to solvent rather than buried inside the protein. Using n.m.r. spectroscopy, we have determined an electron self-exchange rate constant for the b2-cytochrome domain of 2.3 x 10(6) M-1.s-1, which is more than two orders of magnitude larger than the value obtained for microsomal cytochrome b5, a homologue of b2-cytochrome domain.


2021 ◽  
Author(s):  
Kat Pick ◽  
Tracy Lyn Raivio

In this study, we describe the isolation and characterization of novel bacteriophage Kapi1 (vB_EcoP_Kapi1) isolated from a strain of commensal Escherichia coli inhabiting the gastrointestinal tract of healthy mice. We show that Kapi1 is a temperate phage integrated into tRNA argW of strain MP1 and describe its genome annotation and structure. Kapi1 shows limited homology to other characterized prophages but is most similar to the phages of Shigella flexneri, and clusters taxonomically with P22-like phages. Investigation of the lifestyle of Kapi1 shows that this phage displays unstable lysogeny and influences the growth of its host. The receptor for Kapi1 is the lipopolysaccharide O-antigen, and we further show that Kapi1 alters the structure of its hosts O-antigen in multiple ways. We hope to use MP1 and Kapi1 as a model system to explore molecular mechanisms of mammalian colonization by E. coli and ask what the role(s) of prophages in this context might be.


2020 ◽  
Vol 6 (1) ◽  
pp. 13
Author(s):  
Nelson Phiri ◽  
Geoffrey Mainda ◽  
Mercy Mukuma ◽  
Ntazana N. Sinyangwe ◽  
Luke J. Banda ◽  
...  

Objective: Salmonella species and Escherichia coli are major bacterial enteropathogens of worldwide public health importance that cause devastating foodborne diseases, thereby contributing to increased human morbidity and mortality. Both pathogens have also been found to contribute towards the spread of antimicrobial resistance through the food chain, especially in poultry. This study aimed to determine the occurrence of antibiotic-resistant Salmonella spp. and E. coli in broiler chickens at farm level, abattoirs, and open markets in selected districts of Zambia.Methods: A cross-sectional study was undertaken in seven districts of Zambia to determine the resistance profiles of Salmonella spp. and E. coli obtained from broiler chickens at farms, abattoirs, and open markets. A total of 470 samples were collected which include; litter, cloacal swabs, and carcass swabs. Samples were inoculated into buffered peptone water and incubated for 24 hours then sub-cultured onto MacConkey and Xylose Lysine Deoxycholate agar plates. Identification of Salmonella spp. and E. coli was done using the API-20E kit and confirmation by 16S rDNA sequencing. Confirmed isolates were tested against a panel of 09 antibiotics using the Kirby-Bauer disc diffusion method and interpreted according to the Clinical Laboratory Standards Institute guidelines. Data analysis of the antibiotic sensitivity test results was done using WHONET 2018 software.Results: Overall, 4 Salmonella spp. and 280 E. coli were isolated. One of the Salmonella spp. was resistant to ampicillin (25%), amoxicillin/clavulanic acid (25%), and cefotaxime (25%). E. coli antibiotic resistance was highest to tetracycline (81.4%) and 100% susceptibility to imipenem. The antibiotic susceptibility profile revealed 75.7% (237/280) multidrug-resistant (MDR). The highest MDR profile was observed in 8.2% (23/280) isolates in which 6 out of the 9 classes of antibiotics tested were resistant. Out of the 280 isolates, 11.4% (32/280) exhibited Extensive Drug resistance (XDR).Conclusion: The study found antimicrobial resistance to E. coli and Salmonella spp. in market-ready broiler chickens which were resistant to important antibiotics and is of public health concern.


2021 ◽  
Vol 22 (2) ◽  
pp. 223-233
Author(s):  
I.H. Igbinosa ◽  
C. Chiadika

Background: Most Escherichia coli strains are harmless commensals, but some serotypes can cause serious food poisoning in their hosts, and are infrequently responsible for product recalls due to food contamination. The present study was carried out to determine the occurrence of E. coli O157:H7 and other E. coli strains from raw and fermented (nono) milk in Benin City, Nigeria.Methodology: A total of 66 (33 raw and 33 nono) milk samples were obtained from retailers from 3 different stations in Aduwawa market, Benin City, Nigeria between January and June, 2017. Samples were analysed by cultural methods for faecal coliforms using M-Fc agar, E. coli using Chromocult coliform agar, and E. coli O157:H7 using sorbitol MacConkey agar supplemented with cefixime and potassium tellurite. Presumptive E. coli andE. coli O157:H7 isolates were confirmed by polymerase chain reaction (PCR) assay using specific primers. Antimicrobial susceptibility profile of confirmed isolates was performed using the Kirby-Bauer disk diffusion method, with zones of inhibition interpreted according to the guidelines of Clinical and Laboratory Standards Institute (CLSI). Data were  analysed using the SPSS version 21.0.Results: From the 66 nono and raw milk samples assessed in this study, all (100%) were phenotypically positive for E. coli O157:H7. A total of 19 E. coli O157:H7 and 41 other strains of E. coli were confirmed by PCR. The resistance profile of the 19 E. coli O157:H7 isolates showed 100% (19/19) resistance to penicillin G and ampicillin; 94.7% (18/19) to chloramphenicol; 89.5% (17/19) to erythromycin; and 78.9% (15/19) to sulfamethoxazole and oxytetracycline, while the sensitivity profile showed that 100% (19/19) E. coli O157:H7 isolates were sensitive to gentamicin and ofloxacin. The resistance profile of other 41 E. coli isolates showed 100% (41/41) resistance to penicillin G and ampicillin; 97.6% (40/41) to chloramphenicol; and 92.7% (38/41) to erythromycin, while 97.6% (40/41) were sensitive to  gentamicin and kanamycin. Ten E. coli O157:H7 isolates (52.6%) showed extensive drug resistance pattern to 11 antibiotics in 7  antimicrobial classes with multiple antibiotic resistance (MAR) index of 0.46.Conclusion: Findings from the present study clearly indicated that the safety and quality of fresh and fermented milk were not satisfactory and could be of public health concern. Key words: Nono, Escherichia coli; Pathotypes, Resistance index, Public health, Milk


Infectio ◽  
2017 ◽  
Vol 21 (2) ◽  
Author(s):  
Eman Fathi Sharafa ◽  
Iman I. Shabanaa

Shiga toxin-producing Escherichia coli (STEC) strains have emerged as important foodborne pathogens of global public health concern, causing life-threatening diseases. Sheep and their products have been documented as important reservoirs for STECs, especially E. coli O157. The aim of this study was to investigate STECs from diarrheal human and sheep in Al-Madinah Al-Munawarah, Saudi Arabia. Fecal samples were collected between June and August, 2015 from diarrheal humans (n = 134) and sheep (n = 87). Presumptive E. coli human-and sheep-isolated strains were identified for their serotypes, the associated virulence genes (Shiga toxin [stx1 , stx2 ], haemolysin [ehxA] and intimin [eae]) by polymerase chain reaction and their susceptibility to antibiotics. Pulsed-field gel electrophoresis (PFGE) was used to demonstrate the genetic relatedness between Serotype O157:H7 human- and sheep-isolated strains. Forty eight (48/221; 21.7%) STECs were recovered from both human and sheep, their serotypes were as follows: O157:H7, O26:H11, O157:HNM, O26:HNM, O128:H2, O48:HNM, O111:HNM and OUT:HUT. Various virulence profiles and multiple antibiotic resistance were observed among the isolates. Twenty eight O157:H7 serotypes (17 human isolates and 11 sheep isolates) were identified in 13 PFGE pulsotypes, where human and sheep isolates were highly related. PFGE banding profiles together with serotypes and genotypes afford proof that human and sheep can be colonized and infected with similar E. coli O157:H7 strains. Our findings highlight the importance of epidemiological and microbiological surveillance of STECs; as well as the development of control measures to decrease risks associated with zoonotic O157:H7.


2020 ◽  
Author(s):  
Elizabeth Muligisa Muonga ◽  
Geoffrey Mainda ◽  
Mercy Mukuma ◽  
Geoffrey Kwenda ◽  
Bernard Hang'ombe ◽  
...  

Abstract BackgroundAntimicrobial resistance (AMR) of foodborne pathogens is of public health concern, especially in developing countries such as Zambia. This study was undertaken to determine the antimicrobial resistance profiles of Escherichia coli ( E. coli ) and Salmonella isolated from raw retail broiler chicken carcasses purchased from open and supermarkets in Zambia.ResultsA total of 189 E. coli and five Salmonella isolates were isolated. Identification and confirmation of the isolates were done using Analytical Profile Index (API 20E) (Biomerieux ® ) and 16S rRNA sequencing. Antimicrobial susceptibility tests (AST) were performed using the Kirby Bauer disk diffusion technique using a panel of 10 antibiotics. Multiplex PCR was used to determine the presence of three target genes encoding for resistance: tet A, Sul 1 and bla CTX-M . WHONET 2018 software was used to analyse AST results. The E. coli isolates were mostly resistant to tetracycline (79.4%), ampicillin (51.9%), and trimethoprim/sulfamethoxazole (49.7%). Two of the five Salmonella isolates were resistant to at least one antibiotic. Forty- seven (45.2%) of the 104 isolates that were screened for the presence of the resistant genes possessed at least one of the targeted resistance genes.ConclusionThis study has demonstrated the presence of AMR E. coli and Salmonella on raw retail broiler chicken carcasses from open and supermarkets, which is of public health concern.


Author(s):  
Asinamai Athliamai Bitrus ◽  
Peter Anjili Mshelia ◽  
Iliya Dauda Kwoji ◽  
Mohammed Dauda Goni ◽  
Saleh Mohammed Jajere

Antimicrobial resistance has gained global notoriety due to its public health concern, the emergence of multiple drug-resistant bacteria, and lack of new antimicrobials. Extended-spectrum beta-lactamase (ESBL)/ampicillin Class C (AmpC)- producing Escherichia coli and other zoonotic pathogens can be transmitted to humans from animals either through the food chain, direct contact or contamination of shared environments. There is a surge in the rate of resistance to medically important antibiotics such as carbapenem, ESBL, aminoglycosides, and fluoroquinolones among bacteria of zoonotic importance. Factors that may facilitate the occurrence, persistence and dissemination of ESBL/AmpC-Producing E. coli in humans and animal includes; 1). o ral administration of antimicrobials to humans primarily (by physician and health care providers) and secondarily to animals, 2). importation of parent stock and day-old chickens, 3). farm management practice and lack of water acidification in poultry, 4). contamination of feed, water and environment, 5). contamination of plants with feces of animals. Understanding these key factors will help reduce the level of resistance, thereby boosting the therapeutic effectiveness of antimicrobial agents in the treatment of animal and human infections. This review highlights the occurrence, risk factors, and public health importance of ESBL/AmpC-beta-lactamase producing E. coli isolated from livestock.


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