scholarly journals UJI TOKSISITAS ESCHERICHIA COLI ASAL DAGING TERHADAP SEL VERO

2018 ◽  
Vol 18 (2) ◽  
Author(s):  
Widodo Suwito ◽  
Andriani Andriani
Keyword(s):  
Escherichia Coli ◽  
Vero Cell ◽  
Key Words ◽  
Vero Cells ◽  
E Coli ◽  
Traditional Market ◽  
Cattle Faeces

Abstract: Verotoxigenic Escherichia coli (VTEC) is responsible for serious human illnesses. Source of VTEC is cattle faeces which beef contamination. The aims of this study was to determine the ability of E. coli which beef contamination from traditional market to damage the vero cells monolayer. A total of 35 E. coli isolates and vero cells monolayer were used in these study. All isolates E. coli were re-indentified with biochemistry and vero cells monolayer were uesed to determination verotoxigenecity tests. None of E. coli isolates showed damage the vero cells monolayer, so there are not verotoxigenik E. coli. The study showed that all isolate E. coli which beef contamination from tradiotional market none damage the vero cells, so there are not verotoxigenic. Key words: E.coli, beef, vero cell Abstrak: Escherichia coli verotoksigenik (VTEC) menyebabkan penyakit pada manusia. Sumber VTEC adalah feses sapi yang dapat mengkontaminasi daging. Tujuan penelitian ini adalah mengetahui kemampuan E. coli yang diisolasi dari daging sapi di pasar tradisional dalam merusak sel vero monolayer. Sebanyak 35 isolat E. coli dan sel vero monolayer digunakan dalam penelitian ini. Isolat E. coli di identifikasi ulang secara biokimia dan untuk menentukan sifat verotoksigenesitasnya menggunakan sel vero monolayer. Semua isolat E. coli tidak bersifat verotoksigenik karena tidak mampu merusak sel vero. Penelitian ini menunjukkan bahwa E. coli yang mengkontaminasi daging sapi dari pasar tradisional tidak bersifat verotoksigenik.  Kata kunci: E.coli, daging, sel vero. 

Detection of Escherichia coli cytotoxins by enzyme-linked immunosorbent assays

1991 ◽  
Vol 37 (8) ◽  
pp. 650-653 ◽  
Author(s):  
Joan I. Speirs ◽  
Mumtaz Akhtar
Keyword(s):  
Escherichia Coli ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Vero Cell ◽  
Key Words ◽  
E Coli ◽  
Treated Cell ◽  
Cell Extracts ◽  
Immunosorbent Assays ◽  
Key Words Escherichia Coli

Sandwich enzyme-linked immunosorbent assays (ELISAs) were developed to detect Escherichia coli cytotoxins. Wells were coated with monoclonal antibodies from hybridomas 13C4 and (or) 11E10, and biotin conjugates of these antibodies were used for detecting verotoxin 1 and Shiga-like toxin II, respectively. Sensitivities were about 100 and 200 cytotoxic doses, respectively. Verotoxin 2 was detected by ELISA with monoclonal antibody 11E10, but at a sensitivity of only about 4000 cytotoxic doses. ELISA results of polymyxin-treated cell extracts from cultures of 67 E. coli strains were in agreement with Vero cell assay as regards the presence and type of toxin. Key words: Escherichia coli, cytotoxin, ELISA.

Comparison of colony and stool blots for detection of enteropathogens by DNA probes

1991 ◽  
Vol 37 (5) ◽  
pp. 407-410
Author(s):  
Mônica A. M. Vieira ◽  
Beatriz E. C. Guth ◽  
Tânia A. T. Gomes
Keyword(s):  
Escherichia Coli ◽  
Key Words ◽  
Sensitivity And Specificity ◽  
Dna Probes ◽  
Type I ◽  
Enteropathogenic Escherichia Coli ◽  
Key Words Dna ◽  
E Coli ◽  
Heat Stable

DNA probes that identify genes coding for heat-labile type I (LT-I) and heat-stable type 1 (ST-I) enterotoxins, enteropathogenic Escherichia coli adherence factor (EAF), and Shigella-like, invasiveness (INV) are used to evaluate the sensitivity and specificity of stool blots in comparison with the sensitivity and specificity of colony blots in detecting enteropathoghens. The sensitivities of the probes in stool blots are 91.7% for the LT-I probe, 76.9% for the ST-I probes, 78.9% for the EAF probe, and 45.5% for the INV probe. The specificity of all probes is higher than 95%. In general, the stool blot method identifies as many if not more LT-I-, ST-I-, and EAF-producing E. coli infections than the colony blots. Key words: DNA probes, stool blots, enteropathogens, diagnosis.

scholarly journals Assessing the population dynamics of Escherichia coli in a metropolitan river after an extreme flood event

2016 ◽  
Vol 15 (2) ◽  
pp. 196-208 ◽  
Author(s):  
Nicole M. Masters ◽  
Aaron Wiegand ◽  
Jasmin M. Thompson ◽  
Tara L. Vollmerhausen ◽  
Eva Hatje ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Population Dynamics ◽  
Antibiotic Resistance ◽  
Virulence Genes ◽  
Wastewater Treatment Plants ◽  
Vero Cells ◽  
Flood Event ◽  
Typing Method ◽  
E Coli ◽  
Extreme Flood

We investigated Escherichia coli populations in a metropolitan river after an extreme flood event. Between nine and 15 of the 23 selected sites along the river were sampled fortnightly over three rounds. In all, 307 E. coli were typed using the PhP typing method and were grouped into common (C) or single (S) biochemical phenotypes (BPTs). A representative from each of the 31 identified C-BPTs was tested for 58 virulence genes (VGs) associated with intestinal and extra-intestinal E. coli, resistance to 22 antibiotics, production of biofilm and cytotoxicity to Vero cells. The number of E. coli in the first sampling round was significantly (P < 0.01) higher than subsequent rounds, whereas the number of VGs was significantly (P < 0.05) higher in isolates from the last sampling round when compared to previous rounds. Comparison of the C-BPTs with an existing database from wastewater treatment plants (WWTPs) in the same catchment showed that 40.6% of the river isolates were identical to the WWTP isolates. The relatively high number of VGs and antibiotic resistance among the C-BPTs suggests possessing and retaining these genes may provide niche advantages for those naturalised and/or persistent E. coli populations which may pose a health risk to the community.

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.

Prevalence of shiga toxin-producing Escherichia coli O157, O26 and O111 in milk, meat and faeces of cattle, sheep and pigs slaughtered in Benin

2020 ◽  
Vol 152 ◽  
pp. 15667-15675
Author(s):  
Chakirath Folakè Arikè Salifou ◽  
Cyrille Boko ◽  
Isidore Houaga ◽  
Raoul Agossa ◽  
Isabelle Ogbankotan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Virulence Genes ◽  
Animal Origin ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Milk Samples ◽  
Food Of Animal Origin ◽  
Cattle Faeces ◽  
Risk Assessment And Management

Objectives: The study aimed to search for E. coli O157 and non-O157 in milk, meat and faeces of cattle, sheep and pigs slaughtered in Cotonou. Methodology and Results: One hundred and Seventy-Five (175) samples including 25 meat, 25 faeces per species and 25 milk from cattle were analysed for E. coli O157; O26 and O111 and the virulence genes were identified by PCR. The SAS software (1998) and the bilateral Z test were used to calculate and compare the identification frequencies. E. coli O157 was identified in 4% of cattle faeces, 4% of sheep faeces, and 20% of beef and, in 20% of milk samples. E. coli O26 was identified in 12% of cattle faeces and, in 8% of beef samples. E. coli O111 was identified at frequencies of 8%, and 12% in faeces of sheep and pigs, respectively. The eae gene was detected in 4% of beef, ovine meat, milk, pig faeces and in sheep faeces. stx1 was detected in 8% of milk, and in 4% of bovine and sheep faeces. The strains possessing the gene were all of E. coli O157 with the exception of one from pig faeces identified as O111. Conclusions and application of findings: The presence of these serogroups of E. coli with virulence genes poses a real food safety problem in Benin. This study findings must be taken into account for risk assessment and management related to the consumption of food of animal origin. Keywords: Benin, E. coli O157, O26, O111, faeces, meat, milk

Use of a Shiga Toxin (Stx)-Enzyme-Linked Immunosorbent Assay and Immunoblot for Detection and Isolation of Stx-Producing Escherichia coli from Naturally Contaminated Beef

2000 ◽  
Vol 63 (9) ◽  
pp. 1167-1172 ◽  
Author(s):  
HEBA NASHED ATALLA ◽  
ROGER JOHNSON ◽  
SCOTT MCEWEN ◽  
R. W. USBORNE ◽  
C. L. GYLES
Keyword(s):  
Escherichia Coli ◽  
Vero Cell ◽  
Immunosorbent Assay ◽  
Shiga Toxin ◽  
Total Coliform ◽  
Complete Agreement ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spearman Correlation ◽  
Southern Ontario ◽  
E Coli

The purpose of this study was to evaluate an enzyme-linked immunosorbent assay (ELISA) and an immunoblot procedure for detection and isolation of Shiga toxin-producing Escherichia coli (STEC) from beef, and to correlate the presence of STEC in beef with E. coli and total coliform counts. A total of 120 samples of boneless beef supplied to a meat processor in southern Ontario were tested for the presence of STEC, E. coli, and total coliforms. Following enrichment in modified tryptic soy broth, samples were screened for Shiga toxin (Stx) by a Stx-ELISA and a Vero cell assay (VCA). Samples that were positive in the Stx-ELISA were subjected to the Stx-immunoblot for STEC isolation. Overall, 33.3% of samples were positive in the VCA, and 34.2% were positive in the Stx-ELISA. There was almost complete agreement between the Stx-ELISA and the VCA results (kappa = 0.98). The sensitivity and specificity of the Stx-ELISA with respect to the VCA were 100% and 98.75%, respectively. STEC were isolated by the Stx-immunoblot from 87.8% of the samples that were positive in the Stx-ELISA. The STEC isolates belonged to 19 serotypes, with serotype O113:H21 accounting for 10 of 41 isolates. No STEC of serotype O157:H7 were isolated. There was a significant correlation between E. coli counts and total coliform counts (Spearman correlation coefficient = 0.68, P < 0.01). The E. coli count was positively correlated with detection of STEC by both the Stx-ELISA and the VCA (P < 0.01).

The effect of fasting and diet on fecal shedding of Escherichia coli O157:H7 by cattle

2000 ◽  
Vol 80 (4) ◽  
pp. 741-744 ◽  
Author(s):  
S. J. Buchko ◽  
R. A. Holley ◽  
W. O. Olson ◽  
V. P. J. Gannon ◽  
D. M. Veira
Keyword(s):  
Escherichia Coli ◽  
Key Words ◽  
Escherichia Coli O157 ◽  
Fecal Shedding ◽  
E Coli ◽  
Feed Withdrawal ◽  
Alfalfa Silage ◽  
Key Words Escherichia Coli

Cattle naturally infected with Escherichia coli O157:H7 were used to assess the effects of diet and feed withdrawal on the fecal shedding of E. coli O157:H7. Animals were fed an 80% concentrate diet (80% barley and 20% alfalfa silage), fasted for 48 h, fed a 100% forage diet (alfalfa silage), fasted for 48 h, and subsequently re-fed 100% forage (alfalfa silage). There were no differences in the numbers of animals positive for the shedding of E. coli O157:H7 when fed an 80% barley diet or an all-forage diet (P > 0.05) or during the fasting periods following each diet (P > 0.05). Upon re-feeding an all-forage diet following a 48-h fast, animals positive for E. coli O157:H7 shedding increased (P < 0.05), with 42.5% of the animals shedding the pathogen after 5 d. Re-feeding 100% forage following fasting appeared to have increased the number of animals shedding E. coli O157:H7 in their feces, which may have been influenced by diet in addition to fasting. Key words: Escherichia coli O157:H7, fasting, diet, cattle, fecal shedding

scholarly journals Contribution of the Twin Arginine Translocation System to the Virulence of Enterohemorrhagic Escherichia coli O157:H7

2003 ◽  
Vol 71 (9) ◽  
pp. 4908-4916 ◽  
Author(s):  
Nathalie Pradel ◽  
Changyun Ye ◽  
Valérie Livrelli ◽  
Jianguo Xu ◽  
Bernard Joly ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Immunoblot Analysis ◽  
Vero Cells ◽  
Soft Agar ◽  
Enterohemorrhagic Escherichia Coli ◽  
E Coli ◽  
Twin Arginine Translocation ◽  
Tat System ◽  
K 12

ABSTRACT Shiga toxin-producing Escherichia coli O157:H7 is a major food-borne infectious pathogen. In order to analyze the contribution of the twin arginine translocation (TAT) system to the virulence of E. coli O157:H7, we deleted the tatABC genes of the O157:H7 EDL933 reference strain. The mutant displayed attenuated toxicity on Vero cells and completely lost motility on soft agar plates. Further analyses revealed that the ΔtatABC mutation impaired the secretion of the Shiga toxin 1 (Stx1) and abolished the synthesis of H7 flagellin, which are two major known virulence factors of enterohemorrhagic E. coli O157:H7. Expression of the EDL933 stxAB 1 genes in E. coli K-12 conferred verotoxicity on this nonpathogenic strain. Remarkably, cytotoxicity assay and immunoblot analysis showed, for the first time, an accumulation of the holotoxin complex in the periplasm of the wild-type strain and that a much smaller amount of StxA1 and reduced verotoxicity were detected in the ΔtatC mutant cells. Together, these results establish that the TAT system of E. coli O157:H7 is an important virulence determinant of this enterohemorrhagic pathogen.

scholarly journals Evaluation of a Microplate Latex Agglutination Method (Verotox-F Assay) for Detecting and Characterizing Verotoxins (Shiga Toxins) in Escherichia coli

1999 ◽  
Vol 37 (2) ◽  
pp. 396-399 ◽  
Author(s):  
Mohamed A. Karmali ◽  
Martin Petric ◽  
Martina Bielaszewska
Keyword(s):  
Escherichia Coli ◽  
Vero Cell ◽  
Latex Agglutination ◽  
Shiga Toxins ◽  
E Coli ◽  
Cell Assay ◽  
Microtiter Plates ◽  
Culture Filtrates ◽  
Reference Strains

The performance of a commercial microplate latex agglutination assay, the Verotox-F assay, was compared with that of the Vero cell assay for the detection and characterization of Escherichia coli verocytotoxins (VTs). Culture filtrates of 68 VT-positiveE. coli strains (65 human isolates [33 of serotype O157:H7/H−, 32 of non-O157 serotypes] and 3 reference strains) and 104 VT-negative strains (100 human isolates and 4 reference strains) were investigated. The toxin phenotypes and genotypes of the 68 VT-positive isolates were VT1 only (18 strains), VT2 and/or VT2c (33 strains), and VT1 plus VT2 (17 strains). The Verotox-F assay involved incubation of serial dilutions of culture filtrates with equal volumes of latex particles sensitized with anti-VT1 antibody or anti-VT2 antibody in 96-well microtiter plates with appropriate controls and examination for latex agglutination after 20 to 24 h. Compared to the results of the Vero cell assay, the Verotox-F assay was 100% sensitive and 100% specific for the detection of VTs in culture filtrates and correctly identified the toxin types of all 68 VT producers. By checkerboard titration with purified toxins, the sensitivity of the Verotox-F assay was found to be 14 pg (0.7 ng/ml) for VT1, 12 pg (0.6 ng/ml) for VT2, and 350 pg (17.5 ng/ml) for VT2c; this sensitivity is comparable to that of the bioassay. The anti-VT2 latex reagent detected both VT2 and VT2c and did not cross-react with VT1. The anti-VT1 reagent showed a low-level cross-reaction with VT2c only at levels (≥4.5 μg/ml) that were about 1,000-fold higher than those found in culture filtrates. We conclude that the Verotox-F assay is highly sensitive and specific for the detection and characterization of VTs in culture filtrates of human E. coli isolates. The test is rapid, reliable, and easy to perform; its results are easy to interpret; and it should allow testing for VT to become more widely performed.

Colicins G and H and their host strains

1991 ◽  
Vol 37 (10) ◽  
pp. 751-757 ◽  
Author(s):  
D. E. Bradley
Keyword(s):  
Escherichia Coli ◽  
Key Words ◽  
Side Chains ◽  
Human Origin ◽  
Wild Type ◽  
E Coli ◽  
Molecular Weights ◽  
Protein Receptors ◽  
Type Strains ◽  
Host Strains

Escherichia coli strains CA46(pColG) and CA58(pColH) each apparently synthesized two generally similar bactericidal colicin proteins whose molecular weights were approximately 5 500 and 100 000. These proteins were more resistant to trypsin than representative colicins A, D, E1, and V. The smooth wild-type strains harbouring plasmids pColG and pColH were serotyped O169:NM and O30:NM, respectively, being typically associated with nonpathogenic E. coli of human origin. Rough and semirough variants, which were selected using resistance to novobiocin, were intrinsically insensitive to almost as many colicins (10 tested) as their parents. For this reason the wild-type strains would not be useful for identifying colicins G and H on the basis of immunity. The O antigenic side chains of both wild-type strains shielded three of the six bacteriophage protein receptors tested. Key words: colicin, protein, plasmid, O antigen, bacteriophage.

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