Antimicrobial activity of essential oils against Escherichia coli O157:H7 in organic soil

Soil can be a significant source of preharvest contamination of produce by pathogens. Demand for natural pesticides such as essential oils for organic farming continues to increase. We examined the antimicrobial activity of several essential oils against Escherichia coli O157:H7 in soil. Two essential oils (cinnamaldehyde and eugenol), two bio-pesticides (Ecotrol and Sporan) containing essential oils, and an organic acid (acetic acid) at 0.5%, 1.0%, 1.5% and 2.0%, were mixed with organic sandy soil and inoculated with five different strains of E. coli O157:H7. Soils were incubated at room temperature (22 °C) and samples obtained at 1, 7 and 28 days were enumerated to determine survival. E. coli O157:H7 populations in soil were reduced by up to 5 log cfu/g after 24 h incubation at room temperature with 2% cinnamanaldehyde, Ecotrol, Sporan or vinegar. Reduction in E. coli O157:H7 by eugenol was not significantly different from control. Overall, E. coli O157:H7 strain 4406 was the most sensitive of all the five strains tested and cinnamaldehyde was superior to other treatments in reducing E. coli O157:H7 in soil. In general, increases in essential oil concentrations corresponded to reduced survival of E. coli O157:H7 with all oils used in this study. The results suggest that oils can reduce potential contamination of fresh organic produce inadvertently contaminated by soil.     

Antimicrobial activity of mustard essential oil against Escherichia coli O157:H7 and Salmonella typhi

The aim of this study was to investigate how mustard essential oil (EO) affected the cell membrane of Escherichia coli O157:H7 and Salmonella typhi. Intracellular pH and ATP concentration and the release of cell constituents were measured when mustard EO was in contact with E. coli and S. typhi at its minimal inhibitory concentration (MIC) and maximal tolerated concentration (MTC). The treatment with mustard EO affected the membrane integrity of bacteria and induced a decrease of the intracellular ATP concentration. Also, the extracellular ATP concentration increased and a reduction of the intracellular pH was observed in both bacteria. A significantly (P ? 0.05) higher release of cell constituent was observed when both bacteria cells were treated with mustard EO. Electronic microscopy observations showed that the cell membranes of both bacteria were apparently damaged by mustard EO. In conclusion, mustard EO affects the concentration of intracellular component, such as ATP in both bacteria and affects the pH suggesting that cytoplasmic membrane is involved in the antimicrobial action of mustard EO. Mustard EO can be used as an effective antimicrobial agent. We have demonstrated that mustard EO affected the cell membrane integrity, resulting in a loss of cell homeostasis.     

The efficacy of grape seed extract,citric acid and lactic acid on the inactivation of Vibrio parahaemolyticus in shucked oysters

The efficacy of grape seed extract (GE), citric acid (CA) or lactic acid (LA) on the inactivation of Vibrio parahaemolyticus in shucked oysters was studied. The minimum inhibitory concentration (MIC) of GE, CA or LA against V. parahaemolyticus in TSB-1% NaCl was also determined. The shucked oysters were artificially inoculated with V. parahaemolyticus, the inoculated shucked oysters (25 g) were then dipped in solution of GE (0.0, 10.0, 20.0, 50.0, 100, 200, 300 and 500 mg mL⁻¹), CA (0.0, 5.0, 10.0, 15.0, 20.0, 50.0, 100, 200 and 300 mg mL⁻¹) or LA (0.0, 1.0, 5.0, 10.0, 15.0, 20.0, 50.0, 100 and 150 mg mL⁻¹) for 10 min. The population of V. parahaemolyticus in shucked oysters was determined. The MICs of GE, CA or LA against V. parahaemolyticus were 10.0, 5.0 or 1.0 mg mL⁻¹, respectively. A 500, 300 or 150 mg mL⁻¹ GE, CA or LA solutions were needed to reduce the population of V. parahaemolyticus to below the detection level (1.0 log g⁻¹) in shucked oysters.     

Application of grape seed extract in depuration for decontaminating Vibrio parahaemolyticus in Pacific oysters (Crassostrea gigas)

This study investigated the potential application of grape seed extract (GSE) in depuration to increase its efficacy in reducing Vibrio parahaemolyticus populations in Pacific oysters (Crassostrea gigas). Pacific oysters were inoculated with five clinical strains of V. parahaemolyticus to 10⁴⁻⁵ MPN/g and depurated in UV-irradiated artificial seawater (ASW) containing GSE at a level of 1.0% (1.8 mg/mL total phenolic contents as gallic acid equivalents) or 1.5% (3.1 mg/mL total phenolic contents as gallic acid equivalents) at 12.5 °C for up to 5 days. Changes of V. parahaemolyticus populations in oysters during depuration were analyzed every 24 h using the three-tube most probable number (MPN) method. The populations of V. parahaemolyticus in inoculated oysters decreased by 3.06 and 3.71 log MPN/g, respectively, after 4 and 5 days of depuration in ASW at 12.5 °C. However, populations of V. parahaemolyticus in oysters were reduced by 3.01 and 4.18 log MPN/g after 2 days of depuration at 12.5 °C in ASW containing 1.0 and 1.5% GSE, respectively. Further studies confirmed that depuration using ASW containing 1.5% GSE with 3.1 mg/mL total phenolic contents as gallic acid equivalents at 12.5 °C for two days was capable of achieving >3.52 log MPN/g reductions of V. parahaemolyticus in Pacific oysters.     

Synergetic antibacterial efficacy of cold nitrogen plasma and clove oil against Escherichia coli O157:H7 biofilms on lettuce

This study aims to evaluate the antimicrobial effects of cold nitrogen plasma (CNP) and clove oil against Escherichia coli O157:H7 (E. coli O157:H7) biofilm on lettuce. Both clove oil (1 mg/mL, 2 mg/mL and 4 mg/mL) and CNP (400–600 W) displayed significant eradication effect on E. coli O157:H7 biofilms in vitro (p < 0.001). Subsequently, the antibiofilm effect of combined treatment was studied as well. Compared with the respective treatment, combined treatment exhibited remarkable synergistic effect on eradicating E. coli O157:H7 biofilms. The confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) had also visually testified that the antibacterial effects of clove oil on E. coli O157:H7 biofilms (in vitro and on lettuce) were enhanced by CNP at 400 W for short treatment duration. The results of sensory evaluation indicated that combined treatment has mild negative effect on lettuce quality. Moreover, the synergetic antibacterial mechanism of clove oil and CNP against E. coli O157:H7 was concluded as that they could damage the bacterial cell wall and the outer membrane, leading to leakage of cellular components, such as nucleic acid and ATP.     

Antimicrobial activities of red wine-based formulations containing plant extracts against Escherichia coli O157:H7 and Salmonella enterica serovar Hadar

We evaluated the antimicrobial activities of three red wine based plant extract/plant essential oil (EO) formulations: olive extract powder/oregano EO, apple skin extract powder/lemongrass EO, and green tea extract powder/bitter almond EO, and their formula permutations against the foodborne pathogens Escherichia coli O157:H7 and Salmonella enterica serovar Hadar. The following parameters that were expected to affect antimicrobial activity were evaluated: pH (3.6 or 7.0), temperature (4, 22, and 37 °C), incubation time (0, 5, 30, and 60 min) and bacterial load (∼10⁴ or ∼10⁹ bacteria/ml). The formulations with oregano, lemongrass, or bitter almond EO inhibited the growth of both pathogens at acidic pH with bacterial loads of ∼10⁴ or ∼10⁹ bacteria/ml. At neutral pH, the formulations were less effective. Although all the 3-component formulations were fast acting; showing significant activity in less than 5 min, only the activity of the lemongrass-containing formulation markedly increased with longer incubation times from 0 to 60 min. Activity also increased for all 3-component formulations with increasing temperature from 4 °C to 37 °C, although the activity of the lemongrass formulation appeared to level off at 22 °C. Of the tested formulations, the lemongrass EO formulations, at pH 3.6, appeared to be the most effective against the tested pathogens, especially against Salmonella. The most active formulations merit evaluation for antimicrobial efficacy in liquid and solid food.     

Antimicrobial activity of malic acid against Listeria monocytogenes,Salmonella Enteritidis and Escherichia coli O157:H7 in apple,pear and melon juices

Minimal inhibitory (MIC) and minimal bactericidal (MBC) concentrations of malic acid against Listeria monocytogenes, Salmonella Enteritidis and Escherichia coli O157:H7 inoculated in apple, pear and melon juices stored at 5, 20 and 35 °C were evaluated. MICs and MBCs against L. monocytogenes, S. Enteritidis and E. coli O157:H7 were significantly affected by storage temperature, juice characteristics and type of microorganism. Malic acid was more effective at 35 and 20 °C than at 5 °C in all studied fruit juices. E. coli O157:H7 was more resistant to malic acid than S. Enteritidis and L. monocytogenes. Apple, pear and melon juices without malic acid were inhibitory to E. coli O157:H7, S. Enteritidis and L. monocytogenes at 5 °C, whereas, MBCs of 1.5% (v/v) of malic acid in apple and pear juices, and 2% (v/v) in melon juice at 5 °C were needed to reduce E. coli O157:H7, those concentrations being higher than those required to reduce S. Enteritidis and L. monocytogenes in those fruit juices. In addition, concentrations of 2%, 2.5% and 2.5% (v/v) of malic acid added to apple, pear and melon juices, respectively, were required to inactivate the three pathogens by more than 5 log cycles after 24 h of storage at 5 °C. Transmission electron microscopy showed that malic acid produced damage in the cell cytoplasm of pathogens without apparent changes in the cell membrane.     

Influences of vanillin and licorice root extract supplementations on the decimal reduction times of Escherichia coli O157:H7 in mildly heated young coconut liquid endosperm

This study determined the influences of supplementing different combinations of vanillin (0–500 ppm) and licorice root extract (LRE, 0–420 ppm) on the mild heat (55 °C) decimal reduction times (D₅₅-values) of a cocktail of Escherichia coli O157:H7 in young coconut liquid endosperm. The maximum level of supplementation per additive tested was previously established not to result in the lowering of general consumer acceptability of the liquid endosperm. The D₅₅-values ranged from 15.08 to 31.40 min. The influences of both additives on the D₅₅-value fitted significantly into a 2-factor, interaction model with only the joint effect of the additives having significant effect on D₅₅. The combined efficacies of the additives were most significant at concentrations above 250 ppm vanillin and 210 ppm LRE. The results obtained may be used in the establishment of mild heat pasteurization processes for supplemented young coconut liquid endosperm, for better control of quality and safety.     

Comparison of effects of mild heat combined with lactic acid on Shiga toxin producing Escherichia coli O157:H7, O103, O111, O145 and O26 inoculated to spinach and soybean sprout

The aim of the present study was to investigate the effect of lactic acid against Shiga toxin producing Escherichia coli (O157:H7 and non-O157 serogroups including O103, O111, O145 and O26) at different conditions. Soybean sprouts and spinach leaves inoculated with each serogroup of E. coli (∼7.00 + 1.00 log₁₀ cfu/g) were treated with the lactic acid solutions at different concentrations (0% (control), 1.5%, 2.0%, or 2.5%) and at different temperatures (20, 40, or 50 °C) for 3 min. Results indicated that regardless of the treatment temperature, no significant reduction in the numbers of any serogroup occurred in the control group (0%) (p > 0.05). However, lactic acid at concentration of 1.5%, 2% and 2.5% was found to be effective against all organisms tested. There was no significant difference (p > 0.05) between E. coli O157:H7 and non-O157 STEC serogroups at any treatment group. The highest reductions (ca. 4.00 log₁₀ cfu/g) of all serotypes in both produces were observed after immersing into 2.5% lactic acid at 50 °C. The results of this study showed that decontamination of fresh produces such as spinach and soybean sprout with lactic acid solutions prepared at mild temperatures (40 °C and 50 °C) might be an effective safety measure in preventing public health risks associated with these products contaminated with STEC.     

Genotypic characterization of Shiga toxin-producing Escherichia coli O157:H7 isolates in food products from china between 2005 and 2010

Shiga toxin-producing Escherichia coli (STEC) O157 is an important foodborne pathogen. The aims of this study were to determine genetic relatedness of STEC O157 isolated from foods in China. STEC O157 isolates from food were characterized by virulence gene typing, antibiotyping, Multi-locus sequence typing (MLST), pulsed-field gel electrophoresis analysis (PFGE), Multi-locus variable number tandem repeat analysis (MLVA), and the single nucleotide polymorphism (SNP) clade typing. Of the 30 STEC O157 isolates analyzed, all isolates harbored eae, exhA, stx1 and/or stx2 genes with stx2c subtype predominating. By MLST, they were relatively homogenous with only 4 STs. PFGE and MLVA generated 22 pulsotyples and 23 patterns, respectively, which showed considerable diversity. Only one clade 8 isolate was detected. These results indicate that STEC O157 isolates from foods in China were heterogeneous. There was no correlation between genotypic characteristics and sources of isolates. Since different subtyping methods often give different discriminatory powers, the use of more than one subtyping approach is necessary in providing a more accurate picture of the genetic diversity of STEC O157. Four isolates were from ready-to-eat meat or salads, underscores the risk of infections. There is a need for surveillance of STEC O157 in foods and clinically and implementation of prevention strategies to prevent outbreaks of STEC O157 infections in China.     

Assessment of sanitation efficacy against Escherichia coli O157:H7 by rapid measurement of intracellular oxidative stress,membrane damage or glucose active uptake

To reduce cross contamination with foodborne pathogens during washing procedures, sodium hypochlorite (NaOCl) and hydrogen peroxide (H₂O₂) are widely used in the fresh produce industry. To ensure food safety and minimize excessive use of sanitizers and water during sanitation, it is critical to develop a rapid method to assess sanitation efficacy. This study examines the potential for employing oxidative stress, membrane damage and glucose uptake measurements to assess the antimicrobial efficacy of NaOCl (0–70.4 ppm) and H₂O₂ (0–1.6% v/v) toward Escherichia coli O157:H7. The relative amount of intracellular reactive oxygen species (ROS) was measured using Aminophenyl fluorescein (APF) and 2′,7′- dichlorodihydrofluorescein diacetate (H₂DCFDA) for cells treated with NaOCl and H₂O₂ respectively. Results from these ROS sensitive probes revealed a limited correlation between these oxidative stress measurements and inactivation of bacteria measured using the plate counting method. Sanitation experiments, conducted with contaminated pre-cut lettuce leaves in water, were also carried out. Measurement of the bacterial membrane integrity was assessed using the membrane permeable probe propidium iodide (PI) and by evaluating effects on active transport by the glucose transport system using 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG). The results demonstrate that the relative increase in membrane permeability using PI correlated with NaOCl, but did not correlate with H₂O₂ induced reduction of bacterial survival quantified by the plate counting method. In contrast to the other approaches, monitoring the loss of glucose transport system function by measuring the uptake of 2-NBDG displayed strong correlation with the reduction of bacterial survival for both NaOCl and H₂O₂ treatment. Overall, the study demonstrated potential of glucose uptake measurements with 2-NBDG to serve as surrogate method for the traditional plate counting, which is the current gold standard used by the Food and Drugs Administration, to provide a faster analysis of sanitation processes.     

Cloth-based hybridization array system for the identification of Escherichia coli O157:H7

A simple cloth-based hybridization array system (CHAS) utilizing polyester cloth as the solid phase for DNA hybridizations has been adapted as a tool for the identification of presumptive verotoxigenic Escherichia coli O157:H7 colonies isolated from foods by standard culture techniques. Key indicator genes for this organism, rfbO157, fliCH7, vt1 and vt2 (encoding the O and flagellar antigenic determinants and verotoxin genes, respectively) were amplified in a multiplex PCR incorporating digoxigenin-dUTP, followed by hybridization of the amplicons with an array of specific oligonucleotide probes immobilized on polyester cloth, and subsequent immunoenzymatic assay of the bound digoxigenin label. This system includes a simple internal amplification control (IAC) to gauge PCR inhibition based on the incorporation of a primer pair with complementary 3′ ends, resulting in the generation of a unique “primer-dimer” detectable by hybridization with a specific capture probe immobilized on polyester cloth. The CHAS provided sensitive and specific detection of the E. coli O157:H7gene markers, exhibiting the expected patterns of reactivity with a panel of various target and non-target organisms.     

Adherence of cold-adapted Escherichia coli O157:H7 to stainless steel and glass surfaces

The effects of previous cold-induced cell elongation on adherence of Escherichia coli O157:H7 to glass slides and stainless steel surfaces was evaluated at 4 °C for ≤48 h. Planktonic E. coli O157:H7 with and without cold adaptation were prepared at 15 and 37 °C, respectively, and planktonic E. coli O157:H7 containing elongated (>4 ≤ 10 μm) and filamentous (>10 μm) cells were prepared at 6 °C. Despite morphological differences in planktonic E. coli O157:H7 preparations, all three cell types attached to a greater extent to glass than to the stainless steel surfaces. E. coli O157:H7 cells adapted to growth at 15 °C attached better to both glass and stainless steel surfaces (3.2 and 2.6 log cfu/cm², respectively) than cells of the other treatments at ≥24 h. Cells adapted at 6 °C attached to glass slides and stainless steel coupons at levels of 3.0 and 1.8 log cfu/cm², respectively, while E. coli O157:H7 cells grown at 37 °C attached to these surfaces at levels of 2.0 and 1.7 log cfu/cm², respectively. No further attachment of cells from any of the treatments was noted between 24 and 48 h at 4 °C. These results suggest that E. coli O157:H7 cells adapted at 6 °C–15 °C have greater potential to attach to food contact surfaces than those grown at higher temperature. The enhanced biofilm-forming ability of 6 °C or 15 °C-adapted, elongated and filamentous E. coli O157:H7 cells did not appear to be related to the greater entanglement of longer cells within biofilm matrices.     

Prevalence and characterization of Escherichia coli O157:H7 from samples along the production line in Chinese beef-processing plants

This work investigated the prevalence of Escherichia coli O157:H7 and the antibiotic susceptibility, genetic diversity and virulence genes of isolated strains from four beef slaughter plants in China. A total of 510 samples (feces, hide, carcasses and raw meat) were tested for E. coli O157:H7 using enrichment, immunomagnetic separation, and plating on selective media. The prevalence of E. coli O157:H7 in the feces, hide, pre-evisceration carcass, post-evisceration carcass, post-washing carcass, chilled carcass, and raw meat samples was 1.43%, 1.43%, 0%, 0%, 0%, 1.25%, and 0%, respectively. Multiplex PCR assays were used for serotype confirmation and virulence gene detection. stx₂, eaeA and EHEC-hlyA genes were present in all six of the strains, and the stx₁ gene was not present. Fluorescence amplified fragment length polymorphism (FAFLP) was used to determine the genetic diversity of E. coli O157:H7 and revealed a high similarity between the strains isolated from feces and those isolated from carcasses. None of the isolated strains were found to be resistant to sixteen commonly used antimicrobial agents. The results of this study indicate that although E. coli O157:H7 contamination in the Chinese beef industry is sporadic and not as common as reported in other counties, all of the isolates contained three major virulence genes, presenting a high risk of disease for humans. The current research provides baseline information on E. coli O157:H7 prevalence and character profiles in Chinese beef-processing plants that can be used for future studies.     

Role of cellulose in protecting Shiga toxin producing Escherichia coli against oxidative and acidic stress

This study assessed the role of cellulose in protecting Shiga toxin-producing Escherichia coli (STEC) against environmental stress. Cellulose producing STEC strain 19B (O5:H-) and 49B (O103:H2) as well as their cellulose deficient derivates (19D and 49D) were subjected to oxidative (10, 20 and 30 mM H₂O₂) and acidic (pH 3.0, 3.5 and 4.5) stress. Approximately 4.25, 2.46 or 1.74 log CFU ml⁻¹ of the 19B cells survived a 2.5-h treatment with 10, 20 and 30 mM H₂O₂, respectively, while none of the 19D cells (8.02 logs) survived the same treatments. The population of 49D decreased from 7.70 to <0.60 log CFU ml⁻¹, whereas that of 49B decreased from 7.70 to 2.47 log CFU ml⁻¹ by a 2-h treatment with 30 mM H₂O₂. Approximately 2.63 log CFU ml⁻¹ of the 19B cells survived at pH 3.0 for 1 h and 4.75 log CFU ml⁻¹ of the 49B cells survived at pH 3.0 for 1.5 h, but the 19D and 49D cells did not survive these treatments. Results indicated that cellulose protected the selected STEC strains against oxidative treatments with 10-30 mM of H₂O₂ and acidic treatments at pH 3.0-3.5.     

Sublethal injury and recovery of Escherichia coli O157:H7 by high pressure carbon dioxide

A study was carried out to investigate possible sublethal injury caused by high pressure carbon dioxide (HPCD) to Escherichia coli O157:H7 suspended in phosphate-buffered saline (PBS, pH 7.00). The treatment pressure was 5 MPa, treatment temperatures were 25–37 °C and treatment time was 5–70 min. Only 1.21 log₁₀ cycles reduction was obtained in the nonselective medium, while a higher reduction of 5.18 log₁₀ cycles was obtained in the selective medium after treatment at 5 MPa and 25 °C for 50 min, indicating HPCD could induce sublethal injury of E. coli cells and these cells could not recover in the selective medium. The proportion of the sublethally injured E. coli cells in the survivors increased dramatically up to 72.4% with increasing the treatment time from 20 min to 30 min by HPCD at 5 MPa and 25 °C, and this proportion increased gradually with increasing the treatment time, up to 100% at 5 MPa and 25 °C for 60 min. The effect of recovery media, inhibitors of specific metabolic processes, Mg and Ca cations on the recovery of the sublethally injured cells was also investigated. When PBS, peptone water, tryptic soy broth (TSB) and carrot juice were used as recovery media, the maximum repair rate of the sublethally injured cells was found in TSB and the minimum was in PBS. The addition of inhibitors such as sodium azide, chloramphenicol, rifampicin and penicillin G showed that repair of most sublethally injured cells required energy, protein and RNA synthesis, but was not dependent on peptidoglycan synthesis. Moreover, Mg and Ca cations were also needed in the repair.     

Effects of water hardness and pH on efficacy of chlorine-based sanitizers for inactivating Escherichia coli O157:H7 and Listeria monocytogenes

The effects of hardness and pH of water used to prepare electrolyzed oxidizing (EO) water and bleach solutions on the bactericidal activity of sanitizer prepared from the water were examined. EO water and bleach solutions were prepared with hard water of 0, 50, 100, and 200 mg/l as CaCO₃ at pH 5, 6, 7, and 8. Increased water hardness tended to increase free chlorine and oxidation–reduction potential (ORP) and decrease pH of EO water. Chlorine levels also increased with water pH. Water hardness and pH only had minor effect on the pH of bleach solutions. Increasing hardness to 50 mg/l increased antimicrobial effect of EO water against Escherichia coli O157:H7, but reduced when water hardness further increased to 100 mg/l or higher. Water pH had no effect on EO water produced against E. coli O157:H7. Water hardness had no significant effect on bactericidal activity of EO water against Listeria monocytogenes but elevated water pH decreased bactericidal activity of EO water produced against L. monocytogenes. Bleach solution prepared using hard water at 200 mg/l or at pH 7 or higher had significant lower efficacy in inactivating E. coli O157:H7, but had no effect on the inactivation of L. monocytogenes. Results indicate that increasing the hardness or pH of water used to prepare EO water or bleach solutions will decrease the bactericidal activity of sanitizers prepared from the water.     

The influence of acid stress on the growth of Listeria monocytogenes and Escherichia coli O157:H7 on cooked ham

Acid solutions are increasingly being used for decontaminating meat surfaces. On the surfaces of acid-treated meat, the population of microorganism is reduced due to the low pH of acids, and the subsequent growth of the microorganism is reduced due to the residual acids on meat surfaces. Microbial cells on meat surfaces subjected to acid treatments may cross-contaminate untreated meat surfaces, e.g., microorganisms on the surfaces of acid-treated cooked ham cross contaminate the untreated surfaces during slicing. The objective of this study was to examine this scenario in determining the subsequent growth of acid-treated Listeria monocytogenes and Escherichia coli O157:H7 on the surfaces of untreated meat. Cells of multiple-strain L. monocytogenes or E. coli O157:H7 were exposed to HCl solutions of pH 3, 4, or 5 and deionized water at room temperature for 24 h. The acid or deionized water-treated cells were inoculated separately onto cooked ham. Samples inoculated with L. monocytogenes were stored at 4 and 8 °C and samples inoculated with E. coli O157:H7 were stored at 10 and 12 °C. Populations of the pathogens on ham were enumerated during storage, and the lag phase durations (LPD, h) and growth rates (GR, log CFU/h) of the pathogens were determined. The populations of L. monocytogenes and E. coli O157:H7 in pH 5, 4, and 3 solutions were 1.2–3.1 and 0.6–2.4 log CFU/ml, respectively, lower than those in deionized water, indicating an increased acid stress on both microorganisms at lower pHs. L. monocytogenes subjected to pH 3 and pH 4 stresses and E. coli O157:H7 subjected to pH 3 stress exhibited significantly (p < 0.05) extended LPDs and reduced GRs on cooked ham. The growth of L. monocytogenes on ham was more readily reduced by acid stress than that of E. coli O157:H7. This study showed that acid treatments reduced the viability of L. monocytogenes and E. coli O157:H7and the acid stress reduced their subsequent growth ability on untreated ham. Therefore, cross-contamination of L. monocytogenes or E. coli O157:H7 cells from acid-treated meat surfaces onto untreated meat surfaces may not impose increased risk to the product.     

Development of an immunomagnetic separation method for efficient enrichment of Escherichia coli O157:H7

Immunomagnetic separation uses antibody-coated paramagnetic particles to selectively bind and purify the target organisms from a comprehensive range of complex food matrices. Aim of this study was to develop and optimize an immunomagnetic separation method based on monoclonal antibody for efficient isolation of Escherichia coli O157:H7 in food samples. The key parameters for preparing the immunomagnetic beads; the coupling rate between monoclonal antibody and magnetic beads, additive amount of immunomagnetic beads, and magnetic separation times were optimized with different concentrations of E. coli O157:H7. Under optimized conditions, the capture efficiency (CE) was greater than 98% against 10¹–10⁶ cells of E. coli O157:H7 with 0.05 mg immunomagnetic beads. The immunomagnetic beads exhibited high specific binding with E. coli O157:H7 strains (CE > 98%), and low binding with non-target bacteria (CE < 2%) except for S. aureus (CE = 23.6%). The capture efficiency of immunomagnetic beads against E. coli O157:H7 in ground beef and milk samples were 94.4% and 99.8%, respectively.