scholarly journals Detection of phenotypes, virulence genes and phylotypes of avian pathogenic and human diarrheagenic Escherichia coli in Egypt

2016 ◽  
Vol 10 (06) ◽  
pp. 584-591 ◽  
Author(s):  
Hazem Ramadan ◽  
Amal Awad ◽  
Ahmed Ateya
Keyword(s):  
Virulence Genes ◽  
Virulence Gene ◽  
Health Concern ◽  
E Coli ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Shiga Toxin Genes ◽  
Potential Public Health ◽  
Human Stool

Introduction: The purpose from this study was to determine phenotypes, intestinal virulence-associated genes, and phylotypic profiling of human diarrheagenic E. coli (DEC) and avian pathogenic E. coli (APEC). Methodology: A total of 108 chicken visceral organs (liver, spleen, heart) from 36 diseased birds (three organs per each bird) and 78 human stool samples (50 diarrheic patients and 28 healthy persons) were randomly collected during the first half of 2015 in the district of Mansoura city, Egypt. Conventional culturing, serotyping, and molecular characterization of virulence genes and phylogroups were performed. Results: Sixty-five (35%) biochemically identified E. coli isolates were detected from chicken visceral (29/108; 26.9%) and human stool samples (36/78; 46.2%). Serotypes O78, O2, and O1 were the most prevalent serotypes (62%) distinguished from APEC isolates, and only two similar serotypes (O119:H4 and O26:H11) were identified from both APEC and DEC isolates. By polymerase chain reaction (PCR), the respective percentages of 100 and 35 with eae and Shiga toxin genes were detected from APEC isolates while 50%, 27.8%, and 19.4% of human DEC isolates harbored eae, stx1, and stx2 genes, respectively. Phylogrouping revealed a significantly higher occurrence of pathogenic phylogroups (D and B2) in APEC (19/29; 65.5%) than in human DEC isolates (8/36; 22.2%). Conclusions: APEC isolates shared serotypes, virulence genes, and phylotypes with human DEC isolates, which is a subsequent potential public health concern. To the best of our knowledge, this is the first report in Egypt that determines virulence gene and phylogroup coexistence between APEC and DEC isolates.

scholarly journals Molecular Characterization of Plasmid-Mediated Non-O157 Verotoxigenic Escherichia coli Isolated from Infants and Children with Diarrhea

2020 ◽  
Vol 17 (3) ◽  
pp. 0710
Author(s):  
Md Fazlul Karim Khan ◽  
Shah Samiur Rashid
Keyword(s):  
Escherichia Coli ◽  
Virulence Gene ◽  
Multidrug Resistant ◽  
Plasmid Curing ◽  
Health Issues ◽  
E Coli ◽  
Disk Diffusion Assay ◽  
Microbiological Techniques ◽  
Polymerase Chain

A significant increase in the incidence of non-O157 verotoxigenic Escherichia coli (VTEC) infections have become a serious health issues, and this situation is worsening due to the dissemination of plasmid mediated multidrug-resistant microorganisms worldwide. This study aims to investigate the presence of plasmid-mediated verotoxin gene in non-O157 E. coli. Standard microbiological techniques identified a total of 137 E. coli isolates. The plasmid was detected by Perfectprep Plasmid Mini preparation kit. These isolates were subjected to disk diffusion assay, and plasmid curing with ethidium bromide treatment. The plasmid containing isolates were subjected to a polymerase chain reaction (PCR) for investigating the presence of plasmid mediated verotoxin gene (VT1 and VT2) in non-O157 E. coli. Among the 137 E. coli isolates, 49 isolates were non-O157 E. coli while 29 (59.1%) isolates were verotoxin producing non-O157 serotypes and 26 non-O157 VTEC isolates possessed plasmids. Certain isolates harboured single sized plasmid while others had multiple plasmids with different size varied from 1.8kb to 7.6kb. A plasmid containing all (100%) the isolates was multidrug-resistant. Eight isolates changed their susceptibility patterns while three isolates were found to lose plasmid after post plasmid curing treatment and the rest of the isolates (15) remained constant. Different PCR sets characterized 3 plasmid-mediated verotoxins producing non-O157 E. coli. This current study demonstrated the occurrence of plasmid mediated verotoxin gene in non-O157 E. coli. To the best of our knowledge, this is the first report in the global literature on plasmid-mediated verotoxin gene in non-O157 E. coli. Timely diagnosis and surveillance of VTEC infections should prioritize to stop or slow down the virulence gene for dissemination by plasmid-mediated gene transfer amongst the same bacteria or other species.

Virulence and antimicrobial resistance genes associated with the in-vivo pathogenicity of avian pathogenic E. coli isolates

2021 ◽  
Vol 1 (1) ◽  
pp. 17-20
Author(s):  
Ahmed Abd El-Mawgoud ◽  
Azza El-Sawah ◽  
Soad Nasef ◽  
Al-Hussien Dahshan ◽  
Ahmed Ali
Keyword(s):  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Broiler Chickens ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Health Concern ◽  
Pathogenicity Test ◽  
E Coli ◽  
Antimicrobial Resistance Genes

In the current study, ten avian pathogenic E. coli (APEC) isolates of the most predominant APEC serogroups isolated from broiler chickens in Egypt were screened for their virulence and antimicrobial resistance genes pattern using PCR. Five selected virulence gene patterns were further investigated for their in-vivo pathogenicity test. Results showed a 100% prevalence of the β-lactams and tetracyclines resistance genes. However, aminoglycoside and quinolone resistance genes were not detected. Also, 80% of the tested isolates harbored mcr-1 gene, colistin resistance gene. In-vivo pathogenic strains consistently harbored the virulence gene pattern of fimH, fimA, papC, iutA, and tsh. Additionally, the tsh gene was consistently detected with lethal APEC isolates in day-old chicks. These results highlighted the high prevalence of antimicrobial and virulence genes in APEC that potentially represent a public health concern. In this study, the virulence genes fimH, fimA, papC, iutA, and tsh were the most common virulence gene patterns associated with pathogenicity in day-old chicks.

scholarly journals Virulence gene profiles of avian pathogenic Escherichia coli isolated from chickens with colibacillosis in Bulawayo, Zimbabwe

2015 ◽  
Vol 82 (1) ◽  
Author(s):  
Joshua Mbanga ◽  
Yvonne O. Nyararai
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Economic Losses ◽  
Avian Pathogenic Escherichia Coli ◽  
Pcr Assay ◽  
E Coli ◽  
Gene Profiles ◽  
Pathogenic Escherichia Coli ◽  
Polymerase Chain

Colibacillosis, a disease caused by avian pathogenic Escherichia coli (APEC), is one of the main causes of economic losses in the poultry industry worldwide. This study was carried out in order to determine the APEC-associated virulence genes contained by E. coli isolates causing colibacillosis in chickens. A total of 45 E. coli isolates were obtained from the diagnostics and research branch of the Central Veterinary Laboratories, Bulawayo, Zimbabwe. These isolates were obtained from chickens with confirmed cases of colibacillosis after postmortem examination. The presence of the iutA, hlyF, ompT, frz, sitD, fimH, kpsM, sitA, sopB, uvrY, pstB and vat genes were investigated by multiplex polymerase chain reaction (PCR) assay. Of the 45 isolates, 93% were positive for the presence of at least one virulence gene. The three most prevalent virulence genes were iutA (80%), fimH (33.3%) and hlyF (24.4%). The kpsM, pstB and ompT genes had the lowest prevalence, having been detected in only 2.2% of the isolates. All 12 virulence genes studied were detected in the 45 APEC isolates. Virulence gene profiles were constructed for each APEC isolate from the multiplex data. The APEC isolates were profiled as 62.2% fitting profile A, 31.1% profile B and 6.7% profile C. None of the isolates had more than seven virulence genes. Virulence profiles of Zimbabwean APEC isolates are different from those previously reported. Zimbabwean APEC isolates appear to be less pathogenic and may rely on environmental factors and stress in hosts to establish infection.

scholarly journals Genotypical characterization of thermotolerant coliforms isolated from food produced by a Solidarity Economic Venture of Bahia (Brazil)

2020 ◽  
Author(s):  
J. N. Silva ◽  
M. D. Baliza ◽  
F. Freitas ◽  
E. S Cruz ◽  
V. M. A. Camilo ◽  
...  
Keyword(s):  
Virulence Genes ◽  
Food Contamination ◽  
Microbiological Analysis ◽  
Chain Reaction ◽  
E Coli ◽  
Family Farmers ◽  
Two Samples ◽  
Thermotolerant Coliforms ◽  
Polymerase Chain

Abstract Many Solidarity Economic Venture (SEV) are family farmers who seek to add value to production through artisanal processing, which can lead to food contamination. Thus, this study aimed to genotypically characterize thermotolerant coliforms (TtC) strains from food produced by local agribusinesses of SEV during January to April 2019. Samples from thirteen production units (PU) from the SEV were submitted to a microbiological analysis of thermotolerant coliforms (AFNOR 3M1/2 – 09/89), using a fast count method in Petrifilm™ dishes. The Polymerase Chain Reaction (PCR) technique was used to verify the following virulence genes (VGs) associated with Escherichia coli: stx, typical from enterohemorrhagic E. coli (EHEC); bfpA typical from entheropathogenic E. coli (EPEC) and elt and slt, typical from entherotoxigenic E. coli (ETEC). The results showed that two samples of queijadinha (typical Brazilian candy made with eggs and coconut) and one sample of cassava cake presented characteristic colonies TtC. This way, three strains were isolated in order to perform the PCR technique. However, the genes used in the reaction were not detected in the isolated strains. Therefore, it is suggested that the isolated strains are from E. coli pathotypes with different virulence genes than the ones analyzed belong other types of TtC, such as Enterobacter and Klebsiella. Although the virulence of genes has not been confirmed, the presence of TtC on food indicates hygiene flaws during production and, therefore, measurements to control and prevent contamination should be taken.

scholarly journals Prevalence and Virulence Gene Profiles of Escherichia coli O157 from Cattle Slaughtered in Buea, Cameroon

2020 ◽  
Author(s):  
Elvis Achondou Akomoneh ◽  
Seraphine Nkie Esemu ◽  
Achah Jerome Kfusi ◽  
Roland N. Ndip ◽  
Lucy M. Ndip
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Foodborne Pathogen ◽  
Virulence Gene ◽  
Latex Agglutination ◽  
Health Concern ◽  
E Coli ◽  
Uremic Syndrome ◽  
Human Infections ◽  
Pcr Targeting

ABSTRACTBackgroundEscherichia coli O157 is an emerging foodborne pathogen of great public health concern. It has been associated with bloody diarrhoea, haemorrhagic colitis and haemolytic uremic syndrome in humans. Most human infections have been traced to cattle and the consumption of contaminated cattle products. In order to understand the risk associated with the consumption of cattle products, this study sought to investigate the prevalence and identify virulence genes in E. coli O157 from cattle in Cameroon.MethodA total of 512 rectal samples were obtained and analysed using conventional bacteriological methods (enrichment on modified Tryptone Soy Broth and selective plating on Cefixime-Tellurite Sorbitol Mac-Conkey Agar) for the isolation of E. coli O157. Presumptive E. coli O157 isolates were confirmed serologically using E. COLIPRO™ O157 latex agglutination test and molecularly using PCR targeting the rfb gene in the isolates. Characterisation of the confirmed E. coli O157 strains was done by amplification of stx1, stx2, eaeA and hlyA virulence genes using both singleplex and multiplex PCR.ResultsE. coli O157 was detected in 56 (10.9%) of the 512 samples examined. The presence of the virulence genes stx2, eaeA and hylA was demonstrated in 96.4% (54/56) of the isolates and stx1 in 40 (71.4%) of the 54. The isolates exhibited three genetic profiles (I-III) with I (stx1, stx2, eaeA and hlyA) being the most prevalent (40/56; 71.4%) while two isolates had none of the virulence genes tested.ConclusionA proportion of cattle slaughtered in abattoirs in Buea are infected with pathogenic E. coli O157 and could be a potential source of human infections. We recommend proper animal food processing measures and proper hygiene be prescribed and implemented to reduce the risk of beef contamination.

scholarly journals Phylogenetic group determination and plasmid virulence gene profiles of colistin-resistant Escherichia coli originated from the broiler meat supply chain in Bogor, Indonesia

2020 ◽  
Vol 13 (9) ◽  
pp. 1807-1814
Author(s):  
Irma Rahayuningtyas ◽  
Agustin Indrawati ◽  
I Wayan Teguh Wibawan ◽  
Maria Fatima Palupi ◽  
Istiyaningsih Istiyaningsih
Keyword(s):  
Escherichia Coli ◽  
Supply Chain ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Phylogenetic Group ◽  
Health Concern ◽  
Small Scale ◽  
Broiler Meat ◽  
E Coli ◽  
Gene Profiles

Background and Aim: Pathogenic Escherichia coli contamination along the broiler meat supply chain is a serious public health concern. This bacterial infection with multidrug-resistant can lead to treatment failure. Several studies have revealed that avian pathogenic E. coli (APEC) and human extraintestinal pathogenic E. coli (ExPEC) showed a close genetic relationship and may share virulence genes. This study aimed to determine the phylogenetic group and virulence gene profiles in colistin-resistant E. coli obtained from the broiler meat supply chain in Bogor, West Java, Indonesia. Materials and Methods: Fifty-eight archive isolates originated from the cloacal swab, litter, drinking water, inside plucker swab, fresh meat at small scale poultry slaughterhouses, and traditional markets were used in this study. All the isolates were characterized by a polymerase chain reaction to determine the phylogenetic group (A, B1, B2, or D) and virulence gene profiles with APEC marker genes (iutA, hlyF, iss, iroN, and ompT). Results: Phylogenetic grouping revealed that the isolates belong to A group (34.48%), D group (34.48%), B1 group (17.24%), and B2 group (13.79%). The virulence gene prevalence was as follows: iutA (36%), hlyF (21%), ompT (21%), iroN (10%), and iss (9%). The B2 group presented with more virulence genes combinations. iroN, hlyF, and ompT genes were positively associated with the B2 group (p≤0.05). Conclusion: Our results highlight the role of colistin-resistant E. coli originated from the broiler meat supply chain as a potential reservoir for human ExPEC virulence genes.

scholarly journals Molecular Detection, Serotyping, and Antibiotic Resistance of Shiga Toxigenic Escherichia coli Isolated from She-Camels and In-Contact Humans in Egypt

Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1021
Author(s):  
Mohamed Said Diab ◽  
Reda Tarabees ◽  
Yasser F. Elnaker ◽  
Ghada A. Hadad ◽  
Marwa A. Saad ◽  
...  
Keyword(s):  
Antibiotic Susceptibility ◽  
Multiple Drug Resistance ◽  
Significant Risk ◽  
Virulence Gene ◽  
Molecular Techniques ◽  
E Coli ◽  
Milk Samples ◽  
Stool Samples ◽  
Human Stool ◽  
Semi Arid

This study aims to determine the prevalence of STEC in she-camels suffering from mastitis in semi-arid regions by using traditional culture methods and then confirming it with Serological and molecular techniques in milk samples, camel feces, as well as human stool samples for human contacts. In addition, an antibiotic susceptibility profile for these isolates was investigation. Mastitic milk samples were taken after California Mastitis Test (CMT) procedure, and fecal samples were taken from she-camels and human stool samples, then cultured using traditional methods to isolate Escherichiacoli. These isolates were initially classified serologically, then an mPCR (Multiplex PCR) was used to determine virulence genes. Finally, both camel and human isolates were tested for antibiotic susceptibility. Out of a total of 180 she-camels, 34 (18.9%) were mastitic (8.3% clinical and 10.6% sub-clinical mastitis), where it was higher in camels bred with other animals. The total presence of E. coli was 21.9, 13.9, and 33.7% in milk, camel feces, and human stool, respectively, whereas the occurrence of STEC from the total E. coli isolates were 36, 16, and 31.4% for milk, camel feces, and stool, respectively. Among the camel isolates, stx1 was the most frequently detected virulence gene, while hlyA was not detected. The most detected virulence gene in human isolates was stx2 (45.5%), followed by stx1. Camel STEC showed resistance to Oxytetracycline only, while human STEC showed multiple drug resistance to Amoxicillin, Gentamycin, and Clindamycin with 81.8, 72.7, and 63.6%, respectively. Breeding camels in semi-arid areas separately from other animals may reduce the risk of infection with some bacteria, including E. coli; in contrast, mixed breeding with other animals contributes a significant risk factor for STEC emergence in camels.

scholarly journals Detection of Virulence Genes and Biofilm Forming Capacity of Diarrheagenic E. coli Isolated from Different Water Sources

Coatings ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1544
Author(s):  
Sadaf Tariq ◽  
Sobia Tabassum ◽  
Sadia Aslam ◽  
Mika Sillanpaa ◽  
Wahidah H. Al-Qahtani ◽  
...  
Keyword(s):  
Drinking Water ◽  
Water Samples ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Water Sources ◽  
Health Concern ◽  
Pcr Analysis ◽  
E Coli ◽  
Drinking Water Samples ◽  
Different Water Sources

Diarrheagenic Escherichia coli (DEC) are associated with frequent incidences of waterborne infections and pose health risk to individuals who contact water for domestic or recreational uses. Detection of DEC pathotypes in drinking water can be used as an indicator of fecal contamination. This study aimed to investigate the occurrence of DEC pathotypes and their capacity to form biofilms in drinking water samples collected from different water sources. In this study, PCR analysis was used to determine the occurrence of four clinically significant virulence genes of diarrheagenic E. coli, eaeA (Enteropathogenic E. coli), stx1, stx2 (Enterohemorrhagic E. coli) and sth (Enterotoxigenic E. coli), in drinking water samples (n = 35) by using specific primers and conditions. PCR amplicons were visualized by using agarose gel electrophoresis. A total of 12/35 (34%) samples were detected as positive for at least one of the four DEC virulence genes and 11/12 (91%) E. coli isolates harbored virulence gene while 1/12 (8%) E. coli isolates harbored none. The eaeA and sth genes were the most detected genes (75%), while stx1 and stx2 genes were least detected genes (66%). Biofilm assay confirmed that ETEC pathotypes can cause damage in enteric walls by attaching and effacing to persist diarrheal conditions. This study indicated that drinking water of different sources is contaminated with potential DEC pathotypes and it can be a source of diarrheal diseases. The amplification of four virulence genes associated with DEC pathotypes (EPEC, EHEC and ETEC) in drinking water demonstrates that potentially virulent DEC pathotypes are distributed in water sources and may be a cause of health concern. There is, therefore, an urgent need to monitor DEC pathotypes in drinking water.

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