Rapid quantification of Escherichia coli O157:H7 in lettuce and beef using an on-chip staining microfluidic device

Quality assurance is one of the fundamental ways of preventing infections from foodborne pathogens such as Escherichia coli O157:H7, which produces a deadly toxin. Simple, rapid, and accurate methods for the detection of foodborne pathogens are necessary for healthcare management. In the present study, we applied our microfluidic device, which uses a fluorescent staining-based detection system, to enumerate E. coli O157:H7 cells in lettuce and beef samples. E. coli O157:H7 cells spiked into lettuce or beef samples were collected using a 0.2-μm-pore-sized filter or a two-step centrifugation process. The recovery ratios of inoculated E. coli O157:H7 cells from the lettuce and beef samples counted using fluorescence microscopy were 84 (± 10)% and 90 (± 7.3)%, respectively. The counts of E. coli O157:H7 inoculated into lettuce and beef obtained using the microfluidic system were close to the counts obtained using fluorescence microscopy. Our microfluidic approach offers a semi-automated platform for the quantitative detection of microbial cells from complex food samples and facilitates quantification of microbes in food and food production lines within 1 hr.     

A rapid detection of Escherichia coli O157:H7 by competition visual antigen macroarray

Pathogenic bacterial contamination is a serious problem for the food industry and in public health. Rapid, accurate and affordable testing for pathogenic bacterial strains is desirable. In this study, a competition visual antigen macroarray (CVAM) for rapid detection of Escherichia coli O157:H7 (E. coli O157:H7) has been developed. This array was able to utilize an HRP‐labeled anti‐E. coli O157:H7 MAb at a concentration of 1:20000 while having a similar sensitivity of 10 ⁵ CFU/ml for E. coli O157:H7 detection as double antibody sandwich array and conventional ELISA. The detection time of CVAM was short as 2 h and can be examined directly by the naked eye. Moreover, there was no cross‐reactivity in the detection of tested in this study. Application of CVAM for the detection of E. coli O157:H7 artificially contaminated milk samples was feasible that showed the antigen array could be applied in the detection of food pathogen infections.     

CONDITIONS AFFECTING AUTOINDUCER‐2‐BASED QUORUM‐SENSING OF ESCHERICHIA COLI O157:H7 ON FRESH BEEF

ABSTRACT

This study evaluated whether inoculated (none, 1, 5 log colony‐forming units [cfu]/cm²) Escherichia coli O157:H7 would result in detection of autoinducer (AI)‐2‐like activity on beef. Inoculated fresh beef, containing low (LNB) or high (HNB) initial levels of natural flora, was analyzed for bacterial populations and AI‐2‐like activity during aerobic or vacuum‐packaged storage (4, 10, 25C). As expected, no growth of E. coli O157:H7 was detected at 4C, while at 10C, growth was detected only on LNB samples stored aerobically; AI‐2‐like activity was minimal (P ≥ 0.05) at both temperatures. E. coli O157:H7 showed more growth in LNB than HNB, and in aerobically than vacuum‐packaged samples inoculated with 1 log cfu/cm² of the pathogen during storage at 25C. AI‐2‐like activity was generally higher in LNB than HNB samples stored aerobically at 25C, while no significant AI‐2‐like activity was detected in samples stored in vacuum packages. The results indicated that E. coli O157:H7 may exhibit AI‐2‐like activity on aerobically stored beef in the presence of lower initial levels of natural flora, and at temperatures allowing prolific growth of the pathogen. Thus, AI‐2‐based quorum‐sensing of E. coli O157:H7 may not be of importance in beef stored at low temperatures.

PRACTICAL APPLICATIONS

This study presents evidence that Escherichia coli O157:H7 showed autoinducer (AI)‐2 activity and involved in quorum‐sensing on fresh beefcontaining low initial levels of natural flora during aerobic storage at abusive storage temperatures. Thus, AI‐2‐based quorum‐sensing of E. coli O157:H7 may not be important in beef stored at recommended low temperatures.     

Rapid detection of Escherichia coli O157:H7 in milk,bread, and jelly by lac dye coloration‐based bidirectional lateral flow immunoassay strip

Lac dye, a kind of food additive and textile dye, is extracted from lac insect secretions. The ability of lac dye to stain bacteria was firstly found. Based on the discovery, a new bidirectional lateral flow immunoassay strip (BLFIS) for detecting Escherichia coli O157:H7 (E. coli O157:H7) was successfully developed. The BLFIS has two sheets of nitrocellulose (NC) membranes separated by a sample pad. The anti‐E. coli O157:H7 monoclonal antibodies (McAb) were oppositely dispensed on the NC membranes as the control line and test line, respectively. On the side of the control line, a conjugation pad immobilized with pre‐stained E. coli O157:H7 was assembled. The detection limit of BLFIS was 10⁶ CFU/ml. Furthermore, after pre‐incubation in food sample, the detection sensitivity was significantly increased to 100 colonies of the initial bacterial count. This method may be an improvement over traditional colloidal gold lateral flow immunoassay strips, which does not require the pairing of two antibodies to complete the detection, nor the chromogenic nanomaterials to report the detection results, such as gold nanoparticles. And the BLFIS might provide technical support for detecting E. coli O157:H7 in food and ensuring food safety.     

Effectiveness of Active Packaging on Control of Escherichia Coli O157:H7 and Total Aerobic Bacteria on Iceberg Lettuce

Contaminated leafy green vegetables have been linked to several outbreaks of human gastrointestinal infections. Antimicrobial interventions that are adoptable by the fresh produce industry for control of pathogen contamination are in great demand. This study was undertaken to evaluate the efficacy of sustained active packaging on control of Escherichia coli O157:H7 and total aerobic bacteria on lettuce. Commercial Iceberg lettuce was inoculated with a 3‐strain mixture of E. coli O157:H7 at 10² or 10⁴ CFU/g. The contaminated lettuce and un‐inoculated controls were placed respectively in 5 different active packaging structures. Traditional, nonactive packaging structure was included as controls. Packaged lettuce was stored at 4, 10, or 22 °C for 3 wk and sampled weekly for the population of E. coli O157:H7 and total aerobic bacteria. Results showed that packaging structures with ClO₂ generator, CO₂ generator, or one of the O₂ scavengers effectively controlled the growth of E. coli O157:H7 and total aerobic bacteria under all storage conditions. Packaging structure with the ClO₂ generator was most effective and no E. coli O157:H7 was detected in samples packaged in this structure except for those that were inoculated with 4 log CFU/g of E. coli O157:H7 and stored at 22 °C. Packaging structures with an oxygen scavenger and the allyl isothiocyanate generator were mostly ineffective in control of the growth of the bacteria on Iceberg lettuce. The research suggests that some of the packaging structures evaluated in the study can be used to control the presence of foodborne pathogens on leafy green vegetables.     

Detection of E. coli O157:H7 from Ground Beef Using Fourier Transform Infrared (FT-IR) Spectroscopy and Chemometrics

Abstract: FT-IR spectroscopy methods for detection, differentiation, and quantification of E. coli O157:H7 strains separated from ground beef were developed. Filtration and immunomagnetic separation (IMS) were used to extract live and dead E. coli O157:H7 cells from contaminated ground beef prior to spectral acquisition. Spectra were analyzed using chemometric techniques in OPUS, TQ Analyst, and WinDAS software programs. Standard plate counts were used for development and validation of spectral analyses. The detection limit based on a selectivity value using the OPUS ident test was 10⁵ CFU/g for both Filtration-FT-IR and IMS-FT-IR methods. Experiments using ground beef inoculated with fewer cells (10¹ to 10² CFU/g) reached the detection limit at 6 h incubation. Partial least squares (PLS) models with cross validation were used to establish relationships between plate counts and FT-IR spectra. Better PLS predictions were obtained for quantifying live E. coli O157:H7 strains (R²≥ 0.9955, RMSEE ≤ 0.17, RPD ≥ 14) and different ratios of live and dead E. coli O157:H7 cells (R²= 0.9945, RMSEE = 2.75, RPD = 13.43) from ground beef using Filtration-FT-IR than IMS-FT-IR methods. Discriminant analysis and canonical variate analysis (CVA) of the spectra differentiated various strains of E. coli O157:H7 from an apathogenic control strain. CVA also separated spectra of 100% dead cells separated from ground beef from spectra of 0.5% live cells in the presence of 99.5% dead cells of E. coli O157:H7. These combined separation and FT-IR methods could be useful for rapid detection and differentiation of pathogens in complex foods.     

Inactivation of foodborne pathogens in ready-to-eat salad using UV-C irradiation

To examine the applicability of ultraviolet (UV)-C irradiation on the inactivation of foodborne pathogen in ready-to-eat salad, it was inoculated with Escherichia coli O157:H7 and Listeria monocytogenes and then irradiated with UV-C light. Radiation dose required for 90% reduction (d _R) values of E. coli O157:H7 and L. monocytogenes were determined to be 0.21 and 2.48 J/m², respectively. Foodborne pathogen populations significantly (p<0.05) decreased with increasing UV-C irradiation. UV-C irradiation at 8,000 J/m² reduced the populations of E. coli O157:H7 and L. monocytogenes on ready-to-eat salad by 2.16 and 2.57 log CFU/g, respectively.     

Detection of Escherichia coli O157:H7 in Food Using Real-Time Multiplex PCR Assays Targeting the stx 1, stx 2, wzy O157, and the fliC h7 or eae Genes

Escherichia coli O157:H7 is an important foodborne pathogen, and foods of bovine origin and fresh produce have been linked to outbreaks. Real-time multiplex PCR assays were developed to detect E. coli O157:H7 in different foods. Apple cider and raw milk (25 ml) and ground beef and lettuce (25 g) were inoculated with 2 or 20 colony-forming units (CFU) of E. coli O157:H7 380-94 and subjected to enrichment in RapidChek E. coli O157:H7 broth at 42°C. One milliliter of the enrichments was removed at 8 and 20 h, and following DNA extraction, real-time multiplex PCR assays targeting the stx ₁, stx ₂, and wzy _O157 genes in combination with probes and primers targeting either the fliC _h7 or the eae genes were performed using OmniMix HS beads and the SmartCycler. The sensitivity of the real-time multiplex PCR assay was about 225 CFU/PCR. E. coli O157:H7 was detected (fluorescent signal generated for all gene targets) in apple cider, raw milk, lettuce and ground beef samples inoculated with 2 or 20 CFU/g or 25 ml after both 8 and 20 h of enrichment. Enrichments of uninoculated food samples were negative using the multiplex PCR targeting the stx ₁, stx ₂, wzy _O157, and eae genes; however, using the assay targeting the stx ₁, stx ₂, wzy _O157, and fliC _h7 gene combination, a positive result was always obtained for the fliC _h7 gene using uninoculated ground beef enrichments. Use of other primer sets targeting the fliC _h7 gene gave similar results. The real-time multiplex PCR assays targeting the stx ₁, stx ₂, eae, and wzy _O157 or the fliC _h7 genes are sensitive and specific and can be used for the detection of E. coli O157:H7 in food, except that the fliC _h7 gene may not be a suitable target for the detection of E. coli O157:H7 in ground beef.     

Detection and Isolation of Low Levels of E. coli O157:H7 in Cilantro by Real-Time PCR, Immunomagnetic Separation, and Cultural Methods with and without an Acid Treatment

Abstract: Leafy greens such as cilantro, contaminated with Escherichia coli O157:H7, have been implicated in cases of human illnesses. High levels of microflora in fresh cilantro make recovery of low numbers of E. coli O157:H7 difficult. To improve upon current methods, immunomagnetic separation (IMS) techniques in combination with real‐time PCR (RTiPCR) and selective enrichment protocols were examined. Rinsates were prepared from cilantro samples inoculated with low (～0.02 CFU/g) and slightly higher (～0.05 CFU/g) levels of E. coli O157:H7. Rinsate portions were enriched in modified buffered peptone water with pyruvate (mBPWp) for 5 h at 37 °C. After 5 h, selective agents were added to samples and further incubated at 42 °C overnight. Detection and recovery were attempted at 5 and 24 h with and without IMS. IMS beads were screened by RTiPCR for simultaneous detection of stx1, stx2, and uidA SNP. Additionally, broth cultures and IMS beads were streaked onto selective agar plates (Rainbow^®agar, R&F^®E. coli O157 Chromogenic medium, TC‐SMAC and CHROMagar? 0157) for isolation of E. coli O157:H7. Both broth cultures and IMS beads were also acid treated in Trypticase Soy Broth pH 2 prior to plating to selective media to improve upon cultural recovery. Although E. coli O157 strains were detected in most samples by PCR after 5 h enrichment, cultural recovery was poor. However, after 24 h enrichment, both PCR and cultural recovery were improved. Acidification of the broths and the IMS beads prior to plating greatly improved recovery from 24 h enrichment broths by suppressing the growth of competing microorganisms. Practical Application: Detection and recovery of Escherichia coli O157:H7 in fresh produce matrices (e.g., cilantro) can be complicated by high background microflora present in these foods. Rapid detection by molecular methods combined with effective enrichment and isolation procedures such as using immunomagnetic separation (IMS) techniques can quickly identify potential hazards to public health. Additional techniques such as acidification of enrichment broths can exploit acid resistance characteristics of pathogens such as E. coli O157:H7, facilitating their isolation in complex food matrices.     

Rapid and simultaneous quantification of viable Escherichia coli O157:H7 and Salmonella spp. in milk through multiplex real-time PCR

Escherichia coli O157:H7 and Salmonella spp. in milk are 2 common pathogens that cause foodborne diseases. An accurate, rapid, specific method has been developed for the simultaneous detection of viable E. coli O157:H7 and Salmonella spp. in milk. Two specific genes, namely, fliC from E. coli O157:H7 and invA from Salmonella spp., were selected to design primers and probes. A combined treatment containing sodium deoxycholate (SDO) and propidium monoazide (PMA) was applied to detect viable E. coli O157:H7 and Salmonella spp. only. Traditional culture methods and SDO-PMA-multiplex real-time (mRT) PCR assay were applied to determine the number of viable E. coli O157:H7 and Salmonella spp. in cell suspensions with different proportions of dead cells. These methods revealed consistent findings regarding the detected viable cells. The detection limit of the SDO-PMA-mRT-PCR assay reached 10² cfu/mL for Salmonella spp. and 10² cfu/mL for E. coli O157:H7 in milk. The detection limit of SDO-PMA-mRT-PCR for E. coli O157:H7 and Salmonella spp. in milk was significantly similar even in the presence of 10⁶ cfu/mL of 2 nontarget bacteria. The proposed SDO-PMA-mRT-PCR assay is a potential approach for the accurate and sensitive detection of viable E. coli O157:H7 and Salmonella spp. in milk.     

Efficacy of Plant‐Derived Antimicrobials as Antimicrobial Wash Treatments for Reducing Enterohemorrhagic Escherichia Coli O157:H7 on Apples

This study investigated the efficacy of 3 GRAS‐status, plant‐derived antimicrobials (PDAs), trans‐cinnamaldehyde (TC), carvacrol (CR), and β‐resorcylic acid (BR) applied as an antimicrobial wash for killing Escherichia coli O157:H7 on apples. “Red delicious” apples inoculated with a 5 strain mixture of E. coli O157:H7 were subjected to washing in sterile deionized water containing 0% PDA (control), 0.15% TC, 0.35% TC, 0.15% CR, 0.30% CR, 0.5% BR, or 1% BR for 1, 3, and 5 min at 23 °C in the presence and absence of 1% soil, and surviving pathogen populations on apples were enumerated at each specified time. All PDAs were more effective in reducing E. coli O157:H7 compared to the water wash treatment (P < 0.05) and reduced the pathogen by 4‐ to 5‐log CFU/apple in 5 min. Chlorine (1%) was the most effective treatment reducing the pathogen on apples to undetectable levels in 1 min (P < 0.05). Moreover, the antimicrobial effect of CR and BR was not affected by the presence of soil, whereas the efficacy of TC and BR was decreased in the presence of soil. Further, no bacteria were detected in the wash solution containing CR and BR; however, E. coli O157:H7 was recovered in the control wash water and treatment solutions containing TC and chlorine, in the presence of 1% soil (P < 0.05). Results suggest that the aforementioned PDAs, especially CR and BR could be used effectively to kill E. coli O157:H7 on apples when used as a wash treatment. Studies on the sensory and quality characteristics of apples treated with PDAs are needed before recommending their usage.     

Antimicrobial activity of Yerba Mate (Ilex paraguariensis) aqueous extracts against Escherichia coli O157:H7 and Staphylococcus aureus

Abstract: Bioactive compounds from natural plant sources are becoming increasingly important to the food industry. Ilex paraguariensis is used in the preparation of a widely popular tea beverage (Yerba Mate) in the countries of Uruguay, Paraguay, Argentina, and Brazil. In this study, extracts of 4 brands of commercial tea, derived from the holly plant species, Ilex paraguariensis, were evaluated for their ability to inhibit or inactivate bacterial foodborne pathogens. The ultimate goal was to evaluate potential use of the extracts in commercial applications. Dialyzed aqueous extracts were screened for antimicrobial activity against Escherichia coli O157:H7 and Staphylococcus aureus. S. aureus was found to be the more sensitive to extracts than E. coli O157:H7. Minimum bactericidal concentrations (MBCs) were determined to be approximately 150 to 800 μg/mL and 25 to 50 μg/mL against E. coli O157:H7 and S. aureus, respectively. A Uruguayan brand had reduced activity against E. coli O157:H7 compared to the Argentinean brands tested. It was concluded that Yerba Mate could be used as a potential antimicrobial in foods and beverages against these pathogenic bacteria. Practical Application: Soluble extracts from Yerba Mate are natural antimicrobials that can be incorporated into food products to achieve longer shelf life.     

Development of an Effective Treatment for A 5‐Log Reduction of Escherichia coli in Refrigerated Pickle Products

Refrigerated cucumber pickle products cannot be heat processed due to the loss of characteristic sensory attributes. Typically brined refrigerated pickles contain less than 100 mM acetic acid with pH values of 3.7 to 4.0. Refrigeration (4 to 10 °C) helps to inhibit the growth of spoilage bacteria and maintain flavor, texture, and appearance of the pickles. Previous research has shown that pathogenic Escherichia coli strains are unusually acid resistant and survive better in refrigerated acid solutions than at higher temperatures. We found that E. coli O157:H7 can survive for 1 mo or longer at 4 °C in brines typical of commercial refrigerated pickles. Our objective was to develop methods to assure a 5‐log reduction of pathogenic E. coli in these types of products, while maintaining the sensory characteristics. A novel brine formulation was developed, based on current commercial refrigerated pickle brines, which contained 25 mM fumaric acid, 5 mM benzoic acid, 70 mM acetic acid, and 342 mM (2%) sodium chloride, with a pH of 3.8. Sensory data indicate that this formulation did not affect flavor or other sensory attributes of the product, compared to traditional formulations. We achieved a 5‐log reduction of E. coli O157:H7 at 30 °C for 1.52 ± 0.15 d, at 20 °C for 3.12 ± 0.34 d, or at 10 °C for 8.83 ± 0.56 d. Growth of lactic acid bacteria was also inhibited. These results can be used by manufacturers to assure a 5‐log reduction in cell numbers of E. coli O157:H7 and Salmonella without a heat process during the manufacture of refrigerated pickle products. Practical Application : While refrigerated acidified vegetable products are exempt from the acidified foods regulations, we have shown that the vegetative microbial pathogens E. coli O157:H7 can survive for up to 1 mo in these products, given current commercial production practices. To improve the safety of refrigerated pickle products, a brine formulation with reduced acetic acid, but containing fumaric acid, was developed to assure a 5‐log reduction in cell numbers of E. coli O157:H7 without a heat process. The formulation can be used to assure the safety of refrigerated pickled vegetables without altering sensory characteristics.     

Growth inhibition of foodborne pathogens by Oenococcus oeni

Abstract: To explore the possibility of using Oenococcus oeni to inhibit foodborne pathogens, and to characterize antimicrobial compounds produced by O. oeni, 24 strains of O. oeni were tested for their ability to inhibit growth of foodborne pathogens, Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes by using the spot‐on‐lawn method. Of the 24 strains, 17 strains were able to inhibit all 3 pathogens in this study. Proteases, catalase, and buffer solutions were used for determining the type of inhibitory compounds produced from 4 selected strains with stronger inhibitory activity. Antimicrobial activity of 2 strains against the pathogens was completely inactivated by buffer solution, and other 2 strains against E. coli O157:H7 were partially removed. The antimicrobial compound was not sensitive to selected proteases and catalase. Practical Application: There is little information available about using O. oeni for human pathogens control. The results of this study revealed such discovery and potential applications for pathogen control.     

Survival of Escherichia coli O157:H7 during Manufacture and Storage of White Brined Cheese

Escherichia coli O157:H7 is a major foodborne pathogen that causes severe disease in humans. Survival of E. coli O157:H7 during processing and storage of white brined cheese was investigated. Cheeses were prepared using pasteurized milk inoculated with a 4 strain E. coli O157:H7 cocktail (7 log₁₀ CFU/g) with or without yogurt starter culture (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus) and stored in 10% or 15% NaCl brine at 10 and 21 ºC for 28 d. NaCl concentration, water activity (a_w), pH, and numbers of E. coli O157:H7 and lactic acid bacteria (LAB) were determined in cheese and brine. E. coli O157:H7 was able to survive in cheese stored in both brines at 10 and 21 ºC regardless of the presence of starter LAB, although the latter significantly enhanced E. coli O157:H7 reduction in cheese or its brine at 10 ºC. E. coli O157:H7 numbers were reduced by 2.6 and 3.4 log₁₀ CFU/g in cheese stored in 10% and 15% NaCl brine, respectively, in the presence of starter LAB and by 1.4 and 2.3 log₁₀ CFU/g, respectively, in the absence of starter LAB at 10 ºC. The pathogen survived, but at lower numbers in the brines. The salt concentration of cheese stored in 10% brine remained about 5% during ripening, but in 15% brine, the NaCl level increased 1.6% to 8.1% (w/w) by 28 d. Values of pH and a_w slightly decreased 1 d after exposure to brine and reached 5.5 to 6.6 and 0.88 to 0.94, respectively, in all treatments.     

Antimicrobial Effects of Quillaja saponaria Extract Against Escherichia coli O157:H7 and the Emerging Non‐O157 Shiga Toxin‐Producing E. coli

Natural alternate methods to control the spread of Shiga toxin‐producing Escherichia coli (STEC) are important to prevent foodborne outbreaks. Quillaja saponaria aqueous bark extracts (QE), cleared by the U.S. Food and Drug Administration as a natural flavorant, contain bioactive polyphenols, tannins, and tri‐terpenoid saponins with anti‐inflammatory and antimicrobial activity. The objective of this study was to determine the effects of commercial QE against E. coli O157:H7 and non‐O157 strains over 16 h at 37 °C and RT. Overnight cultures of 4 E. coli O157:H7 strains and 6 non‐O157 STECs in Tryptic Soy Broth (TSB) were washed and resuspended in phosphate‐buffered saline (PBS, pH 7.2), and treated with QE and controls including citric acid (pH 3.75), sodium benzoate (0.1% w/w), acidified sodium benzoate (pH 3.75) or PBS for 6 h or 16 h. Recovered bacteria were enumerated after plating on Tryptic Soy Agar, from duplicate treatments, replicated thrice and the data were statistically analyzed. The 4 QE‐treated E. coli O157:H7 strains from initial ～7.5 log CFU had remaining counts between 6.79 and 3.5 log CFU after 16 h at RT. QE‐treated non‐O157 STECs showed lower reductions with remaining counts ranging from 6.81 to 4.55 log CFU after 16 h at RT.  Incubation at 37 °C caused reduction to nondetectable levels within 1 h, without any significant reduction in controls. Scanning electron microscopy studies revealed damaged cell membranes of treated bacteria after 1 h at 37 °C. QE shows potential to control the spread of STECs, and further research in model food systems is needed.     

Susceptibility of Escherichia coli O157 to chitosan‐arginine in beef liquid purge is affected by bacterial cell growth phase

Here, we evaluated the impact of bacterial growth stage on the effect of chitosan‐arginine (Ch‐arg) on Escherichia coli O157:H7 cell numbers and metabolic activity within contaminated beef juice held at room temperature. Using a lux‐marked metabolic reporter strain of E. coli O157:H7, the results showed that Ch‐arg was most bioactive against cells in the lag phase and exponential phase. In comparison, there was a reduced, although still significant, inhibitory effect of Ch‐arg on the viability and metabolic activity of E. coli O157 held in stationary phase. Ch‐arg reduced, but did not eliminate E. coli O157 growth in the meat juice over 48 h. Based on the evidence presented here and elsewhere, we conclude that Ch‐arg can limit the growth and activity of food spoilage bacteria; however, it cannot completely eliminate bacterial contaminants originally present. Ch‐arg should therefore be viewed as a potentially protective measure rather than a biocidal agent that completely eliminates the risk of pathogen transfer in the food chain.     

Growth Inhibitory Effect of Chicken Egg Yolk Antibody (IgY) on Escherichia coli O157:H7

Escherichia coli O157:H7‐specific antibodies (immunoglobulin Y [IgY]) were isolated by the water‐dilution method from the egg yolk of chickens that were immunized with E. coli O157:H7 whole cells. The specific‐binding activity of IgY against E. coli O157:H7 as determined by the enzyme immuno assay showed high levels of activity against bacterial whole cells. IgY binding activity was further demonstrated to have an inhibitory effect on E. coli O157:H7 growth in a liquid medium. The antibacterial function of IgY appeared to result from the interaction of IgY with surface components of E. coli O157:H7, as proven from observation of immunofluorescence and immunoelectron microscopy.     

Production of biofilm and quorum sensing by Escherichia coli O157:H7 and its transfer from contact surfaces to meat,poultry, ready-to-eat deli,and produce products

Multistate outbreaks of Escherichia coli O157:H7 infections through consumption of contaminated foods including produce products have brought a great safety concern. The objectives of this study were to determine the effect of biofilm and quorum sensing production on the attachment of E. coli O157:H7 on food contact surfaces and to evaluate the transfer of the pathogen from the food contact to various food products. E. coli O157:H7 produced maximum levels of AI-2 signals in 12 h of incubation in tested meat, poultry, and produce broths and subsequently formed strong biofilm in 24 h of incubation. In general, E. coli O157:H7 formed stronger biofilm on stainless steel than glass. Furthermore, E. coli O157:H7 that had attached on the surface of stainless steel was able to transfer to meat, poultry, ready-to-eat deli, and produce products. Strong attachment of the transferred pathogen on produce products (cantaloupe, lettuce, carrot, and spinach) was detected (>10³ CFU/cm²) even after washing these products with water. Our findings suggest that biofilm formation by E. coli O157:H7 on food contact surfaces can be a concern for efficient control of the pathogen particularly in produce products that require no heating or cooking prior to consumption.     

Efficacy of Slightly Acidic Electrolyzed Water and UV‐Ozonated Water Combination for Inactivating Escherichia Coli O157:H7 on Romaine and Iceberg Lettuce during Spray Washing Process

Spray washing is a common sanitizing method for the fresh produce industry. The purpose of this research was to investigate the antimicrobial effect of spraying slightly acidic electrolyzed water (SAEW) and a combination of ozonated water with ultraviolet (UV) in reducing Escherichia coli O157:H7 on romaine and iceberg lettuces. Both romaine and iceberg lettuces were spot inoculated with 100 μL of a 3 strain mixture of E. coli O157:H7 to achieve an inoculum of 6 log CFU/g on lettuce. A strong antimicrobial effect was observed for the UV‐ozonated water combination, which reduced the population of E. coli by 5 log CFU/g of E. coli O157:H7 on both lettuces. SAEW achieved about 5 log CFU/g reductions in the bacterial counts on romaine lettuce. However, less than 2.5 log CFU/g in the population of E. coli O157:H7 was reduced on iceberg lettuce. The difference may be due to bacteria aggregation near and within stomata for iceberg lettuce but not for romaine lettuce. The UV light treatment may stimulate the opening of the stomata for the UV‐ozonated water treatment and hence achieve better bacterial inactivation than the SAEW treatment for iceberg lettuce. Our results demonstrated that the combined treatment of SAEW and UV‐ozonated water in the spray washing process could more effectively reduce E. coli O157:H7 on lettuce, which in turn may help reduce incidences of E. coli O157:H7 outbreaks.