Bacteriophage-Based Bioluminescent Bioreporter for the Detection of Escherichia coli O157:H7

2007 ◽  
Vol 70 (6) ◽  
pp. 1386-1392 ◽  
Author(s):  
JENNIFER R. BRIGATI ◽  
STEVEN A. RIPP ◽  
COURTNEY M. JOHNSON ◽  
POLINA A. IAKOVA ◽  
PATRICIA JEGIER ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pathogenic Bacteria ◽  
Vibrio Fischeri ◽  
Homoserine Lactone ◽  
Escherichia Coli O157 ◽  
Recombinant Phage ◽  
E Coli ◽  
Lux Operon ◽  
Multiple Sample ◽  
New Bioassay

The rapid detection of pathogenic bacteria in food and water is vital for the prevention of foodborne illness. In this study, the lux reporter genes were used in a new bioassay that allows pathogen monitoring without multiple sample manipulations or the addition of exogenous substrate. A recombinant phage specific for Escherichia coli O157:H7 was constructed that, upon infection, catalyzes the synthesis of N-(3-oxohexanoyl)-L-homoserine lactone (OHHL). This phage PP01 derivative carries the luxI gene from Vibrio fischeri under the control of the phage promoter PL. OHHL produced by infected E. coli O157:H7 induces bioluminescence in bioreporter cells carrying the V. fischeri lux operon. The ability of phage PP01-luxI to detect several strains of E. coli O157:H7 was confirmed in a 96-well plate assay. In this assay, luxCDABE bioreporter cells capable of detecting OHHL were mixed with phage PP01-luxI and E. coli O157:H7, and luminescence was monitored. Reporter phages induced light in bioreporter cells within 1 h when exposed to 104 CFU/ml of E. coli O157:H7 and were able to detect 10 CFU/ml in pure culture with a preincubation step (total detection time, 4 h). The detection method was also applied to contaminated apple juice and was able to detect 104 CFU/ml of E. coli O157:H7 in 2 h after a 6-h preincubation.

PCR for the Specific Detection of an Escherichia coli O157:H7 Laboratory Control Strain

2015 ◽  
Vol 78 (9) ◽  
pp. 1738-1744 ◽  
Author(s):  
MICHAEL KNOWLES ◽  
DOMINIC LAMBERT ◽  
GEORGE HUSZCZYNSKI ◽  
MARTINE GAUTHIER ◽  
BURTON W. BLAIS
Keyword(s):  
Escherichia Coli ◽  
Pathogenic Bacteria ◽  
Control Measure ◽  
Positive Control ◽  
Food Microbiology ◽  
Escherichia Coli O157 ◽  
Control Strain ◽  
E Coli ◽  
Signature Sequences ◽  
Materials Used

Control strains of bacterial pathogens such as Escherichia coli O157:H7 are commonly processed in parallel with test samples in food microbiology laboratories as a quality control measure to assure the satisfactory performance of materials used in the analytical procedure. Before positive findings can be reported for risk management purposes, analysts must have a means of verifying that pathogenic bacteria (e.g., E. coli O157:H7) recovered from test samples are not due to inadvertent contamination with the control strain routinely handled in the laboratory environment. Here, we report on the application of an in-house bioinformatic pipeline for the identification of unique genomic signature sequences in the development of specific oligonucleotide primers enabling the identification of a common positive control strain, E. coli O157:H7 (ATCC 35150), using a simple PCR procedure.

Inactivation of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella in Cranberry, Lemon, and Lime Juice Concentrates

2003 ◽  
Vol 66 (9) ◽  
pp. 1637-1641 ◽  
Author(s):  
MARA C. L. NOGUEIRA ◽  
OMAR A. OYARZÁBAL ◽  
DAVID E. GOMBAS
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Pathogenic Bacteria ◽  
Antimicrobial Properties ◽  
Escherichia Coli O157 ◽  
Bacterial Inactivation ◽  
Lime Juice ◽  
E Coli ◽  
Log Reduction ◽  
C 32

The production of thermally concentrated fruit juices uses temperatures high enough to achieve at least a 5-log reduction of pathogenic bacteria that can occur in raw juice. However, the transportation and storage of concentrates at low temperatures prior to final packaging is a common practice in the juice industry and introduces a potential risk for postconcentration contamination with pathogenic bacteria. The present study was undertaken to evaluate the likelihood of Escherichia coli O157: H7, Listeria monocytogenes and Salmonella surviving in cranberry, lemon, and lime juice concentrates at or above temperatures commonly used for transportation or storage of these concentrates. This study demonstrates that cranberry, lemon, and lime juice concentrates possess intrinsic antimicrobial properties that will eliminate these bacterial pathogens in the event of postconcentration recontamination. Bacterial inactivation was demonstrated under all conditions; at least 5-log Salmonella inactivation was consistently demonstrated at −23°C (−10°F), at least 5-log E. coli O157:H7 inactivation was consistently demonstrated at −11°C (12°F), and at least 5-log L. monocytogenes inactivation was consistently demonstrated at 0°C (32°F).

Behavior of Escherichia coli O157:H7 on Damaged Leaves of Spinach, Lettuce, Cilantro, and Parsley Stored at Abusive Temperatures

2010 ◽  
Vol 73 (2) ◽  
pp. 212-220 ◽  
Author(s):  
ROWAIDA K. KHALIL ◽  
JOSEPH F. FRANK
Keyword(s):  
Escherichia Coli ◽  
Pathogenic Bacteria ◽  
Leaf Damage ◽  
Escherichia Coli O157 ◽  
Oxidation Reactions ◽  
Leaf Extracts ◽  
Romaine Lettuce ◽  
Growth And Survival ◽  
E Coli ◽  
Additional Information

Recent foodborne illness outbreaks associated with the consumption of leafy green produce indicates a need for additional information on the behavior of pathogenic bacteria on these products. Previous research indicates that pathogen growth and survival is enhanced by leaf damage. The objective of this study was to compare the behavior of Escherichia coli O157:H7 on damaged leaves of baby Romaine lettuce, spinach, cilantro, and parsley stored at three abusive temperatures (8, 12, and 15°C). The damaged portions of leaves were inoculated with approximately 105 CFU E. coli O157:H7 per leaf. The pathogen grew on damaged spinach leaves held for 3 days at 8 and 12°C (P < 0.05), with the population increasing by 1.18 and 2.08 log CFU per leaf, respectively. E. coli O157:H7 did not grow on damaged Romaine leaves at 8 or 12°C, but growth was observed after 8 h of storage at 15°C, with an increase of less than 1.0 log. Growth of E. coli O157:H7 on Romaine lettuce held at 8 or 12°C was enhanced when inocula were suspended in 0.05% ascorbic acid, indicating the possibility of inhibition by oxidation reactions associated with tissue damage. Damaged cilantro and Italian parsley leaves held at 8°C for 4 days did not support the growth of E. coli O157:H7. Behavior of the pathogen in leaf extracts differed from behavior on the damaged tissue. This study provides evidence that the damaged portion of a leafy green is a distinct growth niche that elicits different microbial responses in the various types of leafy greens.

Efficacy of Chemical Treatments in Eliminating Salmonella and Escherichia coli O157:H7 on Scarified and Polished Alfalfa Seeds

2001 ◽  
Vol 64 (10) ◽  
pp. 1489-1495 ◽  
Author(s):  
SARAH L. HOLLIDAY ◽  
ALAN J. SCOUTEN ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Chemical Treatment ◽  
Organic Material ◽  
Pathogenic Bacteria ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Mechanical Abrasion ◽  
Chemical Treatments ◽  
The Impact ◽  
Alfalfa Seeds

Alfalfa seeds are sometimes subjected to a scarification treatment to enhance water uptake, which results in more rapid and uniform germination during sprout production. It has been hypothesized that this mechanical abrasion treatment diminishes the efficacy of chemical treatments used to kill or remove pathogenic bacteria from seeds. A study was done to compare the effectiveness of chlorine (20,000 ppm), H2O2 (8%), Ca(OH)2 (1%), Ca(OH)2 (1%) plus Tween 80 (1%), and Ca(OH)2 (1%) plus Span 20 (1%) treatments in killing Salmonella and Escherichia coli O157:H7 inoculated onto control, scarified, and polished alfalfa seeds obtained from two suppliers. The influence of the presence of organic material in the inoculum carrier on the efficacy of sanitizers was investigated. Overall, treatment with 1% Ca(OH)2 was the most effective in reducing populations of the pathogens. Reduction in populations of pathogens on seeds obtained from supplier 1 indicate that chemical treatments are less efficacious in eliminating the pathogens on scarified seeds compared to control seeds. However, the effectiveness of chemical treatment in removing Salmonella and E. coli O157:H7 from seeds obtained from supplier 2 was not markedly affected by scarification or polishing. The presence of organic material in the inoculum carrier did not have a marked influence on the efficacy of chemicals in reducing populations of test pathogens. Additional lots of control, scarified, and polished alfalfa seeds of additional varieties need to be tested before conclusions can be drawn concerning the impact of mechanical abrasion on the efficacy of chemical treatment in removing or killing Salmonella and E. coli O157:H7.

Inactivation of Pathogenic Bacteria by Cucumber Volatiles (E,Z)-2,6-Nonadienal and (E)-2-Nonenal†

2004 ◽  
Vol 67 (5) ◽  
pp. 1014-1016 ◽  
Author(s):  
M. J. CHO ◽  
R. W. BUESCHER ◽  
M. JOHNSON ◽  
M. JANES
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Bactericidal Activity ◽  
Pathogenic Bacteria ◽  
Brain Heart Infusion ◽  
Escherichia Coli O157 ◽  
Infusion Solution ◽  
E Coli ◽  
Log Reduction

The effects of (E,Z)-2,6-nonadienal (NDE) and (E)-2-nonenal (NE) on Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium were investigated. A suspension of each organism of 6 to 9 log CFU/ml was incubated for 1 h at 37° C in brain heart infusion solution that contained 0 to 500 or 1,000 ppm of NDE or NE. Depending on concentration, exposure to either NDE or NE caused a reduction in CFU of each organism. Treatment with 250 and 500 ppm NDE completely eliminated viable B. cereus and Salmonella Typhimurium cells, respectively. L. monocytogenes was the most resistant to NDE, showing only about a 2-log reduction from exposure to 500 ppm for 1 h. Conversely, this concentration of NDE caused a 5.8-log reduction in E. coli O157:H7 cells. NE was also effective in inactivating organisms listed above. A higher concentration of NE, 1,000 ppm, was required to kill E. coli O157:H7, L. monocytogenes, or Salmonella Typhimurium compared with NDE. In conclusion, both NDE and NE demonstrated an apparent bactericidal activity against these pathogens.

scholarly journals Isolation and Identification of Escherichia coli O157:H7 Lytic Bacteriophage from Environment Sewage

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Tessa Sjahriani ◽  
Eddy Bagus Wasito ◽  
Wiwiek Tyasningsih
Keyword(s):  
Escherichia Coli ◽  
Host Specificity ◽  
Pathogenic Bacteria ◽  
Statistical Tests ◽  
Plaque Morphology ◽  
Escherichia Coli O157 ◽  
Lytic Bacteriophage ◽  
Foodborne Disease ◽  
Isolation And Identification ◽  
E Coli

Escherichia coli O157:H7 is one of the pathogenic bacteria causing foodborne disease. The use of lytic bacteriophages can be a good solution to overcome the disease. This study is aimed at isolating lytic bacteriophages from environmental sewage with E. coli O157:H7 bacterial cells. The sample used in this study was eight bacteriophages, and the technique used in identifying E. coli O157:H7 carriers of the stx1 and stx2 genes was PCR. The double layer plaque technique was used to classify bacteriophages. Plaque morphology, host specificity, and electron micrograph were used to identify the bacteriophages. The result obtained plaque morphology as a clear zone with the largest diameter size of 3.5 mm. Lytic bacteriophage could infect E. coli O157:H7 at the highest titer of 10 × 10 8   PFU / mL . Bacteriophages have been identified as Siphoviridae and Myoviridae. Phage 3, phage 4, and phage 8 could infect Atypical Diarrheagenic E. coli 1 (aDEC1) due to their host specificity. The Friedman statistical tests indicate that lytic bacteriophage can significantly lyse E. coli O157:H7 ( p = 0.012 ). The lysis of E. coli O157:H7 by phage 1, phage 2, phage 3, and phage 5 bacteriophages was statistically significant, according to Conover’s posthoc test ( p < 0.05 ). The conclusion obtained from this study is that lytic bacteriophages from environmental sewage could lyse E. coli O157:H7. Therefore, it could be an alternative biocontrol agent against E. coli O157:H7 that contaminates food causing foodborne disease.

scholarly journals Transcriptomic Analysis of Viable but Non-Culturable Escherichia coli O157:H7 Formation Induced by Low Temperature

2019 ◽  
Vol 7 (12) ◽  
pp. 634 ◽  
Author(s):  
Junliang Zhong ◽  
Xihong Zhao
Keyword(s):  
Escherichia Coli ◽  
Low Temperature ◽  
Differentially Expressed Genes ◽  
Pathogenic Bacteria ◽  
Differentially Expressed ◽  
Transmembrane Transport ◽  
Pathway Enrichment Analysis ◽  
Escherichia Coli O157 ◽  
Vbnc State ◽  
E Coli

Escherichia coli O157:H7 is one of the most common pathogenic bacteria that pose a threat to food safety. The aim of this study was to investigate the mechanisms of the formation of viable but non-culturable (VBNC) E. coli O157:H7 induced by low temperature (−20 °C) using RNA sequencing (RNA-Seq) transcriptomics analysis. The results of the present investigation revealed the presence of 2298 differentially expressed genes in VBNC cells, accounting for 46.03% of the total number of genes. Additionally, GO function and KEGG pathway enrichment analysis were performed to investigate the functional and related metabolic pathways of the differentially expressed genes. We found that the ion transport, protein synthesis, and protein transmembrane transport activities were significantly improved in the VBNC cells, indicating that E. coli O157:H7 cells synthesized a considerable amount of protein to maintain the levels of their functional metabolic processes and life activities in the VBNC state. In conclusion, we suggest that the increased synthesis of proteins such as SecY, FtsY, and Ffh might indicate that they are the key proteins involved in the improvement of the transmembrane transport activities in VBNC E. coli O157:H7 cells, maintaining their functional metabolism in the VBNC state and enhancing their survival ability under low temperatures.

Proteome Analysis of Virulence Factor Regulated by Autoinducer-2–like Activity in Escherichia coli O157:H7

2007 ◽  
Vol 70 (2) ◽  
pp. 300-307 ◽  
Author(s):  
YOUNGHOON KIM ◽  
SANGNAM OH ◽  
EUN YOUNG AHN ◽  
JEE-YOUNG IMM ◽  
SEJONG OH ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Proteome Analysis ◽  
Protein Translation ◽  
Escherichia Coli O157 ◽  
Dependent Manner ◽  
Two Dimensional Gel Electrophoresis ◽  
E Coli ◽  
Autoinducer 2

Many pathogenic bacteria, including Escherichia coli O157:H7, can control gene expression in a cell density–dependent manner by producing small signaling molecules (autoinducers) in a process known as quorum sensing. In this study, the effects of the autoinducer-2–like activity on the expression of proteins, including virulence factors, in E. coli O157:H7 were characterized by proteomic analysis. Compared with the control, E. coli O157:H7 strains in the presence of autoinducer-2–like activity exhibited elevated virulence by more rapidly forming cell aggregates on epithelial cells and rapidly killing the nematode Caenorhabditis elegans, the surrogate host. Two-dimensional gel electrophoresis revealed 18 proteins that were upregulated by autoinducer-2–like activity and 4 proteins that were down-regulated. These proteins were further characterized by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry and are involved in the metabolic process, adaptation and protection, cell motility, secretion, envelope biogenesis, and protein translation. These results indicate that the newly identified proteins are associated with the control of virulence in E. coli O157:H7 and that these proteins can be potential targets for the development of antibiotics and other antimicrobial agents.

Efficacy of Cetylpyridinium Chloride in Immersion Treatment for Reducing Populations of Pathogenic Bacteria on Fresh-Cut Vegetables

2001 ◽  
Vol 64 (12) ◽  
pp. 2071-2074 ◽  
Author(s):  
HONG WANG ◽  
YANBIN LI ◽  
MICHAEL F. SLAVIK
Keyword(s):  
Escherichia Coli ◽  
High Performance Liquid Chromatography ◽  
Salmonella Typhimurium ◽  
High Performance ◽  
Pathogenic Bacteria ◽  
Room Temperature ◽  
Cetylpyridinium Chloride ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Fresh Cut

The efficacy of cetylpyridinium chloride (CPC) immersion to reduce the numbers of three pathogenic bacteria (Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7) on three different fresh-cut vegetables (broccoli, cauliflower, and radishes) was studied. The fresh-cut vegetables were inoculated with one of the three pathogenic bacteria at a concentration of 105 CFU/ml for 1 h at room temperature and then treated with 0.1 or 0.5% CPC immersion for 1 min. Both Salmonella Typhimurium and E. coli O157:H7 plates were incubated from 48 to 72 h at 37°C, and L. monocytogenes plates were incubated from 72 to 96 h before being counted. The results of three experiments showed that for the average of the three vegetables treated with 0.1 and 0.5% CPC, L. monocytogenes was reduced by 2.85 and 3.70 log CFU/g, Salmonella Typhimurium by 2.37 and 3.15 log CFU/g, and E. coli O157:H7 by 1.01 and 1.56 log CFU/g, respectively, in comparison with the vegetables treated with water only. The 0.5% CPC treatment was significantly different (P &lt; 0.05) from the 0.1% CPC treatment on reduction of L. monocytogenes, Salmonella Typhimurium, and E. coli O157:H7. The CPC residual on the treated vegetables and their washing solutions were evaluated by using high-performance liquid chromatography.

Death of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in Garlic Butter as Affected by Storage Temperature

2002 ◽  
Vol 65 (12) ◽  
pp. 1976-1980 ◽  
Author(s):  
BARBARA B. ADLER ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Pathogenic Bacteria ◽  
Storage Time ◽  
Storage Temperature ◽  
Rapid Decline ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Pathogen Populations ◽  
And Storage

Garlic is known to have antimicrobial activity against several spoilage and pathogenic bacteria. However, the fate of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in garlic butter has not been reported. This study was undertaken to determine the viability of these organisms in garlic butter as affected by the type of raw minced garlic added to the butter, storage temperature, and storage time. Unsalted butter at 40°C was combined with raw minced jumbo, elephant, or small-cloved garlic at a 4:1 butter/garlic ratio (wt/wt), inoculated with mixed-strain suspensions of Salmonella, E. coli O157:H7, or L. monocytogenes, and stored at 4.4, 21, or 37°C for up to 48 h. All pathogens retained their viability at 4.4°C, regardless of the presence of garlic. The addition of garlic to butter enhanced the rates of inactivation of all three pathogens at 21 and 37°C. The most rapid decline in pathogen populations was observed at 37°C. The inactivation of L. monocytogenes occurred more slowly than did that of Salmonella or E. coli O157:H7. The inactivation of Salmonella and L. monocytogenes was more rapid in jumbo garlic butter than in elephant or small-cloved garlic butter. It is concluded that Salmonella, E. coli O157:H7, and L. monocytogenes did not grow in unsalted butter, with or without garlic added (20%, wt/wt), when inoculated products were stored at 4.4, 21, and 37°C for up to 48 h.

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