Rapid identification of Escherichia coli O157:H7 in bovine feces using the antibody-direct epifluorescent filter technique (Ab-DEFT)

1996 ◽  
Vol 51 (3-4) ◽  
pp. 343-349 ◽  
Author(s):  
M.L. Tortorello ◽  
D.S. Stewart ◽  
W.C. Cray
Keyword(s):  
Escherichia Coli ◽  
Rapid Identification ◽  
Escherichia Coli O157 ◽  
Filter Technique ◽  
Direct Epifluorescent Filter Technique ◽  
Bovine Feces

Antibody-Direct Epifluorescent Filter Technique and Immunomagnetic Separation for 10-h Screening and 24-h Confirmation of Escherichia coli O157:H7 in Beef

1996 ◽  
Vol 59 (10) ◽  
pp. 1072-1075 ◽  
Author(s):  
LAWRENCE RESTAINO ◽  
H. JEFFREY CASTILLO ◽  
DIANA STEWART ◽  
MARY LOU TORTORELLO
Keyword(s):  
Escherichia Coli ◽  
Membrane Filtration ◽  
Immunomagnetic Separation ◽  
Rapid Screening ◽  
Epifluorescence Microscopy ◽  
Objective Lens ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Filter Technique ◽  
Direct Epifluorescent Filter Technique

Rapid screening of beef for the presence of Escherichia coli O157:H7 was shown to be feasible using a 10-h enrichment in modified buffered peptone water and the antibody-direct epifluorescent filter technique (Ab-DEFT). The Ab-DEFT involved membrane filtration, fluorescent antibody staining and epifluorescence microscopy and was accomplished in less than 1 h. The procedure allowed detection of the pathogen artificially inoculated into beef patties at 0.1 CFU/g. The 10-h nonselective enrichment broth supported rapid growth, which provided sufficient numbers of cells for a positive determination by the Ab-DEFT after fewer than 10 microscope fields were scanned using a 40× objective lens. Immunomagnetic separation using anti-E. coli O157 Dynabeads® was used to confirm presumptively positive cultures within 24 h. The ease and rapidity of the Ab-DEFT may provide a substantial time and cost savings to the beef industry for screening beef for the presence of E. coli O157:H7.

scholarly journals Factors Involved in the Persistence of a Shiga Toxin-Producing Escherichia coli O157:H7 Strain in Bovine Feces and Gastro-Intestinal Content

2018 ◽  
Vol 9 ◽  
Author(s):  
Audrey Segura ◽  
Pauline Auffret ◽  
Delphine Bibbal ◽  
Marine Bertoni ◽  
Alexandra Durand ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Intestinal Content ◽  
Escherichia Coli O157 ◽  
Bovine Feces

Sensitivity of bacteriologic culture for detection of Escherichia coli O157:H7 in bovine feces.

1995 ◽  
Vol 33 (10) ◽  
pp. 2616-2619 ◽  
Author(s):  
M W Sanderson ◽  
J M Gay ◽  
D D Hancock ◽  
C C Gay ◽  
L K Fox ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Escherichia Coli O157 ◽  
Bovine Feces

scholarly journals Development and Characterization of a Fluorescent-Bacteriophage Assay for Detection of Escherichia coli O157:H7

1999 ◽  
Vol 65 (4) ◽  
pp. 1397-1404 ◽  
Author(s):  
Lawrence Goodridge ◽  
Jinru Chen ◽  
Mansel Griffiths
Keyword(s):  
Escherichia Coli ◽  
Flow Cytometry ◽  
Immunomagnetic Separation ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Lower Detection ◽  
Unreliable Method ◽  
Direct Epifluorescent Filter Technique ◽  
Bacterial Concentrations

ABSTRACT In this paper we describe evaluation and characterization of a novel assay that combines immunomagnetic separation and a fluorescently stained bacteriophage for detection of Escherichia coliO157:H7 in broth. When it was combined with flow cytometry, the fluorescent-bacteriophage assay (FBA) was capable of detecting 104 cells/ml. A modified direct epifluorescent-filter technique (DEFT) was employed in an attempt to estimate bacterial concentrations. Using regression analysis, we calculated that the lower detection limit was between 102 and 103cells/ml; however, the modified DEFT was found to be an unreliable method for determining bacterial concentrations. The results of this study show that the FBA, when combined with flow cytometry, is a sensitive technique for presumptive detection of E. coliO157:H7 in broth cultures.

Specific Identification of Escherichia coli O157 by an Immunostick Method Using Commercially Available Antibodies

1996 ◽  
Vol 59 (6) ◽  
pp. 670-673 ◽  
Author(s):  
SHIU W. HUANG ◽  
TSUNG C. CHANG
Keyword(s):  
Escherichia Coli ◽  
Rapid Identification ◽  
Escherichia Coli O157 ◽  
Commercial Preparation ◽  
E Coli ◽  
Specificity And Sensitivity ◽  
Sandwich Enzyme Immunoassay ◽  
Antigen Antibody ◽  
Phosphate Buffered Saline ◽  
Somatic Antigens

A sandwich enzyme-immunoassay performed on plastic sticks was developed to specifically identify Escherichia coli O157. Colonies of test bacteria grown on tryptic soy agar were suspended in phosphate-buffered saline, heated in a 100°C water bath for 15 min, and incubated with the plastic sticks coated with a commercial preparation of anti-E. coli O157 antibodies at 37°C for 1.5 h. After incubation, the same antibodies labeled with peroxidase were used to produce the signal of antigen-antibody reaction. For 35 strains of E. coli O157 (among them 34 were E. coli O157:H7) tested, all produced strong reactions by the immunoassay. For 162 strains of E. coli with somatic antigens other than O157 and 38 strains of other genera tested, only one strain (Salmonella bietri) produced a false-positive reaction. The specificity and sensitivity of the immunostick assay were 100% (35/35) and 99.5% (199/200), respectively. The detection limit of the assay for E. coli O157:H7 (CCRC 15991) was about 105 CFU/ml. The method, which can be carried out within 3 h, is useful for rapid identification of suspect E. coli O157 isolated on selective media.

Thermal Inactivation of Escherichia coli O157:H7 Isolated from Ground Beef and Bovine Feces, and Suitability of Media for Enumeration

1998 ◽  
Vol 61 (3) ◽  
pp. 285-289 ◽  
Author(s):  
M. ROCELLE S. CLAVERO ◽  
LARRY R. BEUCHAT ◽  
MICHAEL P. DOYLE
Keyword(s):  
Escherichia Coli ◽  
Thermal Inactivation ◽  
Ground Beef ◽  
Treatment Temperature ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Polyethylene Bags ◽  
D Values ◽  
Bovine Feces

Rates of thermal inactivation of five strains of Escherichia coli O157:H7 isolated from ground beef implicated in outbreaks of hemorrhagic colitis and five strains isolated from bovine feces were determined. Ground beef (22% fat, 10 g), inoculated with individual test strains at populations ranging from 6.85 to 7.40 log10 CFU g−1 of beef, was formed into patties (0.3 cm thick and 8.0 cm in diameter) and sealed in polyethylene bags. For each strain and treatment temperature (54.4, 58.9, 62.8, 65.6, or 68.3°C), 6 bags were simultaneously immersed into a recirculating water bath. Viable cells in patties heated for various lengths of time were enumerated by plating diluted samples on sorbitol MacConkey agar supplemented with 4-methylumbelliferyl-β-d-glucuronide (MSMA) and modified eosin methylene blue (MEMB) agar. Regardless of strain or treatment temperature, higher numbers of E. coli O157:H7 cells were generally recovered on MEMB agar than on MSMA, indicating the inferiority of MSMA as a recovery medium for quantitative determination of E. coli O157:H7 cells in heat-processed ground beef. Significantly (P ≤ 0.05) higher D values when enumeration was done using MEMB agar compared with MSMA. Mean D values for combined strain data at 54.4, 58.9, 62.8, and 65.6°C from cultures on MEMB agar were 123.90, 6.47, 0.62, and 0.20 min, respectively, whereas D values of 25.5, 5.21, 0.57, and 0.18 min were obtained at the same temperatures from cultures on MSMA. Results suggest that cooking ground beef patties to an internal temperature of 68.3°C for 40 s will inactivate at least 99.99% of E. coli O157:H7 cells; z values of 4.0 and 5.1°C were calculated from mean D values obtained from MEMB agar and MSMA, respectively, as recovery media. Differences in D values and z values existed among strains but rates of thermal inactivation do not appear to be correlated with the sources of the isolates.

scholarly journals Efficacy of Octenidine Hydrochloride for Reducing Escherichia coli O157:H7, Salmonella spp., and Listeria monocytogenes on Cattle Hides

2012 ◽  
Vol 78 (12) ◽  
pp. 4538-4541 ◽  
Author(s):  
Sangeetha Ananda Baskaran ◽  
Abhinav Upadhyay ◽  
Indu Upadhyaya ◽  
Varunkumar Bhattaram ◽  
Kumar Venkitanarayanan
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Escherichia Coli O157 ◽  
Salmonella Spp ◽  
Content Type ◽  
Presence And Absence ◽  
Bovine Feces

ABSTRACTThe efficacy of octenidine hydrochloride (OH; 0.025, 0.15, and 0.25%) for inactivatingEscherichia coliO157:H7,Salmonellaspp., andListeria monocytogeneson cattle hides was investigated at 23°C in the presence and absence of bovine feces. All tested concentrations of OH were effective in decreasing more than 5.0 log CFU of bacteria/cm2in 5 min (P< 0.01). The results suggest that OH could be used to decontaminate cattle hides; however, further studies under commercial settings are necessary to validate these results.

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