Survival of bacteria in less than thorough cooked,brine-injected steaks

Bacteria that may include enteric pathogens are introduced into muscle tissues when they are injected with brine. Consequently, steaks of such non-intact, non-comminuted tissues should be cooked to temperatures sufficient to render them microbiologically safe. Studies were conducted to identify the minimum safe temperature for cooking those meats. Suspensions of Escherichia coli and Listeria innocua at numbers about 8 log cfu/ml, in broth or in brines containing 2% or 5% of each of NaCl and sodium tripolyphosphate that were not supplemented or were supplemented with 2% soy protein or 2% emulsified sunflower oil were injected into the centres of 3 cm thick steaks. Groups of four steaks injected with each type of suspension were cooked to temperatures at the centres of 50, 55, 60, 65 or 70 °C. Bacteria were recovered from samples from the centres of steaks, on agars that did or did not allow resuscitation of injured cells of each organism. The results indicated that E. coli but not L. innocua was injured by the brines; that supplementing brines with protein or oil did not protect the organisms from injury or inactivation by heating; and that cooking to a central temperature > 60 ? 65 °C was sufficient to inactivate all bacteria in the meat. The centres of steaks were subsequently injected with 0.1 ml portions of five strain cocktails of E. coli O157:H7 or Listeria monocytogenes in broth containing a dye and the bacteria at numbers >8 log cfu/ml. Steaks were cooked to temperatures of 63, 64 or 65 °C, with holding for 0, 1 or 2 min after cooking before excision of all the dyed tissue from each steak and enumeration of bacteria from the tissue on resuscitating media. The results indicated that cooking to 65 °C without holding would be sufficient to reduce numbers of E. coli O157:H7 or L. monocytogenes in non-intact, non-comminuted steaks by ?7 log units.     

Molecular screening and characterization of Shiga toxin-producing Escherichia coli in retail foods

Shiga toxin-producing Escherichia coli (STEC) strains are one of the most important recently emerged groups of food borne pathogens. This study investigated the prevalence of molecular markers for STEC and characteristics of E. coli O157 isolates from foods sold at retail markets in Wuhan, China. A total of 489 samples (350 meat products and 139 raw vegetables) were purchased from 22 large scale markets between July of 2011 and September of 2013. The meat samples consisted of frozen chicken products, raw pork, raw beef, frozen fish products and processed duck products. The raw vegetable samples consisted of lettuce, bok choy, radish, spinach, cucumber, and tomato. Shiga toxin genes (stx1 and stx2) and an O-group marker of the seven main pathogenic STEC serogroups (O157, O26, O45, O103, O111, O121, and O145) were detected in the samples by using PCR. 100% agreement was obtained between the results of the PCR targeting for wzy_O157 and the PCR targeting for rfbE_O157 gene. The result demonstrated that PCR assay targeting for wzy_O157 gene can be employed as an effective screening method for E. coli O157 in food sample. In the study, E. coli O157 and non-O157 STEC were detected in 55 (11.2%) and 75 (15.3%) samples by PCR screening, respectively. There was significant difference in the occurrence of STEC contamination between supermarkets (19/127, 15.0%) and open markets (111/362, 30.7%) (P < 0.05). Out of 489 samples, 5 samples carried O45, 1 sample carried O145 and 1 sample carried O111. Markers for O103, O26 and O121 were not detected. This result differed from other reports. Immunomagnetic separation based cultivation technique was used to isolate E. coli O157 from 27 food samples collected in 2013. Finally 7 E. coli O157 isolates were obtained. Among the 7 isolates, the prevalent stx genotype was stx_1a and stx_2a. Four E. coli O157 strains exhibited toxic effects on Vero cells, while 3 isolates had no detectable cytotoxicity effects even though they contained stx genes. All E. coli O157 isolates were sensitive to the 12 antimicrobials tested except for roxithromycin. There are some inconsistencies between the PCR screening and culture results. Characteristics of STEC isolates should be evaluated and considered for monitoring STEC contamination in foods.     

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.     

Inhibitory effect of Cinnamomum cassia oil on non-O157 Shiga toxin-producing Escherichia coli

Shiga toxin-producing Escherichia coli (STEC) have caused numerous foodborne outbreaks. Compared with the most well-known STEC E. coli O157:H7, importance of non-O157 STEC has been underestimated and they have gained far less attention till increasing outbreaks recently. Using natural plant materials as antimicrobial agents is a heated area. Therefore in this study, Cinnamomum cassia, a widely used spice in cuisine, was tested for its antibacterial efficacy on CDC “top six” non-O157 STECs including O26, O45, O103, O111, O121, O145. Gas chromatography-mass spectrometry analysis showed that the major component of C. cassia oil was cinnamaldehyde (59.96%). The disk diffusion assay indicated that 20 μL 4% (v/v) C. cassia oil per disk resulted in inhibition zones of 15.0 mm, 18.5 mm, 15.7 mm, 19.3 mm, 18.8 mm, and 25.3 mm for O26:H11, O45:NM, O103:H2, O111:H2, O121:H19, and O145:NT, respectively. Minimum inhibitory concentration for all tested non-O157 STECs were 0.025% (v/v). Minimum bactericidal concentration was strain dependent, which was 0.05% (v/v) for O26:H11, O121:H19, O145:NT, while 0.1% (v/v) for O45:NM, O103:H2 and O111:H2. Growth kinetics showed that at the low inoculation of approximate 2.5 × 10⁵ CFU/mL, C. cassia oil at the concentration of 0.01875% (v/v) completely inhibited the growth of O26:H11 and O145:NT for at least 24 h, and increased the duration of lag phase of O45:NM, O103:H2, O111:H2, O121:H19 by18, 12, 6, and 16 h, respectively. Including 0.025% (v/v) C. cassia oil completely inhibited the growth of all tested non-O157 STECs for at least 24 h. At high inoculation of 5 × 10⁶ CFU/mL, inhibition effect of C. cassia oil decreased. Death curve showed that including as low as 0.05% (v/v) C. cassia oil could kill non-O157 STECs. 0.1% (v/v) C. cassia oil showed bactericidal effects on all tested non-O157 STECs within 15 min. C. cassia oil at the concentration of 0.15% (v/v) killed all O26:H11, O121:H19 and O145:NT within 30 min, while O45:NM, O103:H2 and O111:H2 at 120, 60, and 60 min, respectively. In conclusion, C. cassia oil can effectively inhibit the growth of non-O157 STECs at concentration as low as 0.025% (v/v). Our data suggest that C. cassia oil has the potential to be used as a natural antibacterial agent in food industry.     

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.     

Reuterin,lactoperoxidase, lactoferrin and high hydrostatic pressure on the inactivation of food-borne pathogens in cooked ham

The antimicrobial effect of high hydrostatic pressure (HHP) processing combined with reuterin, lactoperoxydase system (LPS) and lactoferrin (LF) on the survival of Listeria monocytogenes, Salmonella enterica subsp. enterica serovar Enteritidis and Escherichia coli O157:H7 in sliced cooked ham stored under strict refrigeration temperature (4 °C) and mild temperature abuse conditions (10 °C) was investigated. One day after treatment, L. monocytogenes counts in HHP at 450 MPa for 5 min were 0.8 log units lower, but a recovery was observed with counts not significantly different to those observed in control after 35 d. S. Enteritidis and E. coli O157:H7 levels were reduced around 5 log cfu/g by the pressure treatment (450 MPa/5 min) and the numbers of these pathogens did not increase significantly during the 35 d of storage at 4 °C. The individual application of reuterin and LPS influenced the survival of the three pathogens studied, extending the lag phase of L. monocytogenes and diminishing S. Enteritidis and E. coli levels throughout storage, whereas no effect was recorded when LF was added. When reuterin or LPS were applied in combination with HHP there was a synergistic antimicrobial effect against L. monocytogenes, avoiding at 4 °C the recovery observed with individual treatments. These combined treatments also kept the levels of S. Enteritidis and E. coli O157:H7 below the detection limit (<1 log unit) in cooked ham stored at 4 and 10 °C during 35 d. The results obtained in the present work suggest that HHP at 450 MPa for 5 min in combination with LPS or reuterin would be useful as a hurdle technology approach against L. monocytogenes, S. Enteritidis and E. coli O157:H7 in cooked ham.     

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.     

Growth characteristics of Shiga toxin-producing Escherichia coli (STEC) stressed by chlorine,sodium chloride,acid, and starvation on lettuce and cantaloupe

Shiga toxin-producing Escherichia coli (STEC) is a major foodborne pathogen causing serious illnesses and hospitalizations in the United States. Bacteria that are exposed to environmental stresses during food processing may exhibit different growth patterns in the subsequent growth environment. The purpose of this study was to examine the effect of environmental stresses on the growth of O15H and non-O157 STEC in lettuce or cantaloupe. Strains of O157:H7 and non-O157 STEC (O26:H11, O103:H1, O104:H4, and O145:NM) were subjected to four selected stresses including 2 ppm of chlorine, a_w of 0.97 (osmotic stress), and pH 5 (acid stress) at 22 °C for 24 h, or starvation (lack of nutrients) at 22 °C for 15 d. A cocktail mix of stressed or non-stressed (control) O157 or non-O157 at 3 log CFU/g (control or stressed) was inoculated on lettuce or cantaloupe and incubated at 10 and 22 °C for four weeks. While there were significant differences (p < 0.05) in the growth of stressed and unstressed cells of non-O157 STEC, no difference was observed in the growth of stressed and unstressed O157 STEC cells. Osmotic-stressed non-O157 STEC had significantly higher cell populations than control with 2 log difference (9.0 vs. 6.8 log CFU/g) at 10 °C on lettuce and 1 log difference (9.3 vs. 8.3 log CFU/g) at 22 °C on cantaloupe after 4 weeks. Acid-stressed non-O157 STEC had significantly higher cell populations than control at 10 °C after 4 weeks with >1 log difference (7.7 vs. 6.3 log CFU/ml) on cantaloupe. Starvation-stressed non-O157 STEC showed significantly higher cell populations than control with 1 log difference (8.4 vs. 7.2 log CFU/g) at 22 °C on cantaloupe after 4 weeks. The results indicated that osmotic, acid, or starvation stress may enhance the growth of non-O157 STEC on lettuce or cantaloupe and lead to a greater safety risk.     

Occurrence,serotypes and virulence genes of non-O157 Shiga toxin-producing Escherichia coli in fresh beef,ground beef,and beef burger

This study determined the prevalence, serotypes and virulence genes distribution of non-O157 Shiga toxin-producing Escherichia coli in meat products collected from butchers shops and supermarkets in Mansoura city, Egypt. We have characterized 18 non-O157 STEC strains among the identified 100 E. coli isolates recovered from the examined 87 meat product samples. The prevalence of non-O157 STEC strains in fresh beef, ground beef and beef burger samples were 11.1% (3/27), 16.7% (5/30), and 33.3% (10/30), respectively. The eighteen non-O157 STEC isolated strains were serotyped into seven (38.9%) O111:H8, six (33.3%) O26:H11, two (11.1%) O111:H–, and one (5.56%) for each of O55:H7, O126:H5 and O128:H2. PCR assays for different virulence genes showed that nine (50%), eleven (61.1%), and nine (50%) strains carry stx1, stx2, and eae genes, respectively. The distribution of shiga toxin genes among the isolated strains indicated that seven (38.9%) strains harbored stx1 only, nine (50%) strains harbored stx2 only, and two (11.1%) strains harbored both stx1 and stx2. The eae gene was present in association with five (27.8%), three (16.7%), and one (5.6%) strains that harbored stx1 only, stx2 only, and both stx1 and stx2, respectively. This study concluded that the examined meat products, particularly beef burger, consumed in Egypt are considerably contaminated with a variety of non-O157 STEC serotypes, and hence consumption of such products may constitute a potential health risk for consumers.     

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 ascorbic acid alone or in combination with lactic acid on Escherichia coli O157:H7 in laboratory medium and carrot juice

The antimicrobial effects of ascorbic acid alone and in combination with lactic acid, against Escherichia coli O157:H7 in Brain Heart Infusion (BHI) broth and in carrot juice as a food model was investigated. In control samples, E. coli O157:H7 continued to growth from 3.98 ± 0.2 log CFU and reached to 8.88 ± 0.1 log₁₀ CFU after 8 h at 37° C; however, bacterial population was undetectable level in BHI in the presence of 0.4% ascorbic acid and 0.2% lactic acid. In carrot juice, E. coli O157:H7 continue to grow from initial population count of 4.41 ± 0.9 log₁₀ CFU to 8.75 ± 0.07 log₁₀ CFU at 37 °C for 24 h. Similarly, the bacteria population was undetectable when 0.2 or 0.4% ascorbic acid and 0.2% lactic acid applied. Our findings suggest the application of ascorbic acid, in combination with lactic acid, may have potential as preservative to inhibit the growth of E. coli O157:H7 in food.     

Effect of organic acids,hydrogen peroxide and mild heat on inactivation of Escherichia coli O157:H7 on baby spinach

Minimally processed baby spinach contaminated with Escherichia coli O157:H7 has been associated with multiple outbreaks of foodborne illnesses recently. Chlorinated water is widely used to wash vegetables commercially, but this washing procedure has limited efficacy and can lead to the formation of carcinogenic substances. This study was conducted to determine the effects of organic acids and hydrogen peroxide alone and in binary combinations with or without mild heat (40 and 50 °C) on the inactivation of Escherichia coli O157:H7 on baby spinach. Baby spinach leaves were dip-inoculated with E. coli O157:H7 to a level of 6 log CFU/g and stored at 4 °C for 24 h before treatment. Individual washing solutions (1% and 2% lactic acid [LA], citric acid [CA], malic acid [MA], tartaric acid [TA], acetic acid [AA], hydrogen peroxide [H₂O₂] as well as binary combinations of LA, CA, MA and H₂O₂ at final concentrations of 1% were used to decontaminate spinach leaves at 22, 40 or 50 °C for 2–5 min to test their efficacy in reducing E. coli O157:H7. Chlorinated water (200 ppm free chlorine) decreased the population of E. coli O157:H7 on baby spinach by only 1.2–1.6 log CFU/g, which was not significantly different from DI water washing. Washing with 1% LA at 40 °C for 5 min was the most effective treatment achieving a 2.7 log reduction of E. coli O157:H7 which is significantly higher than chlorine washing. Washing with LA + CA or LA + HP at 40 °C for 5 min was equally effective against E. coli O157:H7, resulting in a 2.7 log reduction of E. coli O157:H7. The application of mild heat significantly enhanced the efficacy of washing solutions on the inactivation of E. coli O157:H7. There was, however, no significant difference between treatments at 40 °C for 5 min and 50 °C for 2 min. The results suggested that the use of organic acids in combination with mild heat can be a potential intervention to control E. coli O157:H7 on spinach.     

Pulsed electric field inactivation of E. coli O157:H7 and non-pathogenic surrogate E. coli in strawberry juice as influenced by sodium benzoate,potassium sorbate,and citric acid

Current FDA regulations require that juice processors achieve a 5 log CFU/ml reduction of a target pathogen prior to distributing products. Whereas thermal pasteurization reduces the sensory characteristics of juice, pulsed electric field (PEF) treatments can be conducted at lower temperatures and may preserve sensory characteristics.Escherichia coli O157:H7 (ATCC 43895) and a non-pathogenic E. coli (ATCC 35218), respectively, were inoculated into single-strength strawberry juice with or without 750 ppm sodium benzoate (SB), 350 ppm potassium sorbate (PS), and 2.7% citric acid (CA). Juice was treated at outlet temperatures of 45, 50 and 55 °C at a field strength of 18.6 kV/cm for 150 μs with a laboratory-scale PEF unit. Inactivation of surrogate E. coli at 45, 50, and 55 °C were 2.86, 3.12, and 3.79 log CFU/ml, respectively, in plain juice (pH 3.4), and 2.75, 3.52, and 5.11 with the addition of benzoic and sorbic acids (pH 3.5). Inactivation of E. coli O157:H7 under the same conditions were 3.09, 4.08, and 4.71 log CFU/ml, respectively, and 2.27, 3.29, and 5.40 with antimicrobials. E. coli O157:H7 in juice with antimicrobials and 2.7% CA (pH 2.7) treated with PEF was reduced by 2.60, 4.32 and 6.95 log CFU/ml at 45, 50 and 55 °C while the surrogate E. coli decreased by 3.54, 5.69, and 7.13 log under the same conditions. When juice (pH 2.7) was held for 6 h without PEF treatment, higher numbers of E. coli 35218 (7.17 log CFU/ml) were inactivated than of acid-resistant E. coli O157:H7 (3.89 log). Slightly greater PEF inactivation of E. coli O157:H7 than of the surrogate bacterium indicates that E. coli ATCC 35218 provides a margin of safety when used as a surrogate for O157:H7 in plain strawberry juice or in juice + SB + PS at 45–50 °C, or with SB + PS and CA at 55 °C.     

Characteristics of coliphage ECP4 and potential use as a sanitizing agent for biocontrol of Escherichia coli O157:H7

Escherichia coli O157:H7 is an important pathogenic bacterium to humans because it produces various toxins, such as shiga-toxin. Coliphage ECP4, which belongs to the Siphoviridae family, was isolated from bovine feces to test its utility as a potential agent for the biocontrol of E. coli O157:H7. The burst size of coliphage ECP4 was about 80 PFU/cell, after a latent period of 30–35 min. Coliphage ECP4 was susceptible to temperatures above 70 °C; however, its stability was slightly reduced to 1–2 log PFU/ml after 30 min in 70% ethanol. In addition, the shiga toxin gene was not detected on coliphage ECP4. Coliphage ECP4 inhibited the growth of E. coli O157:H7 in vegetable juice, and was not detected in cabbage after 5 h. When coliphage ECP4 was applied to biofilm-formed E. coli O157:H7, E. coli O157:H7 was efficiently reduced. The newly identified coliphage ECP4 might effectively reduce E. coli O157:H7 or its biofilmed-form. Therefore, the coliphage ECP4 might be an efficient sanitizer for fresh produce contaminated with E. coli O157:H7 in the biofilm environment.     

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.     

Escherichia coli O26:H11 growth in co-culture with an antagonistic strain of Hafnia alvei according to pH and temperature

This study examines the combined effects of temperature (33 °C, 27 °C, 22 °C, 18 °C) and pH (5.3, 5.8, 6.3, 6.8), within the range of values encountered during cheese-making, on the growth of an Escherichia coli O26:H11 strain (21765) co-cultivated with an antagonistic strain of Hafnia alvei (B16) at different initial concentrations (4, 5, 6, 7 Log CFU/mL). Experimental designs including these parameters were applied to a liquid medium (Brain Heart Infusion) and to uncooked pressed model cheeses. In both these environments, the polynomial models gave a good prediction of E. coli O26:H11 levels after 6–24 h of incubation. In both predictive models, ANOVA analysis showed that temperature and pH both had a major positive effect on the level of E. coli O26:H11 at 6 h and the initial concentration of H. alvei had a negative effect. In BHI medium and in uncooked pressed model cheese at 24 h, temperature had a weaker positive effect when B16 was inoculated at 7 Log rather than 5 Log CFU/mL. In cheese at 24 h, pH also had a more marked positive effect on E. coli O26:H11 levels when H. alvei was inoculated at 7 Log. The predictive model established in BHI gave a good prediction of the levels of E. coli O26:H11 in uncooked pressed cheeses at 6 h but overestimated them at 24 h. H. alvei B16 inoculation at 7 Log CFU/mL associated with pH 5.3 and a temperature of 22 °C may help to control E. coli O26:H11 at 24 h in cheeses. Further studies with cheese production will be necessary to manage pH (via starter culture) in combination with temperature, without modifying the sensorial qualities of the cheese.     

Sublethal injury and recovery of Escherichia coli O157:H7 and K-12 after exposure to lactic acid

Lactic acid can induce sublethal injury of Escherichia coli. When conditions become favorable, injured E. coli can recover physiological function and fully virulence, which is a great concern in the field of food safety. The injury and recovery of E. coli O157:H7 and K-12 by lactic acid were investigated in this study. Sublethally injured E. coli cells widely persisted after a 60-min exposure to lactic acid with different pH values (E. coli O157:H7: pH 3.0–4.6, E. coli K-12: pH 3.4–5.0). The sublethally injured ratio of E. coli O157:H7 and K-12 by lactic acid decreased as incubation temperature decreasing. Both sublethally injured E. coli O157:H7 and K-12 induced by lactic acid could be completely recovered in trypticase soy broth at 37 °C within 60 min. For both E. coli O157:H7 and K-12, sodium pyruvate, Tween 80, or certain cations (Mn, Fe, or Zn) could significantly increase the recovery ratio. The recovery ratios of injured E. coli K-12 and O157:H7 were only 74.2% and 92.6% after 80 min incubation in minA, respectively. But they can both be completely recovered within 80 min in minA with sodium pyruvate, with Tween 80, or with cations (Mn, Fe, or Zn). But Mg and Ca cations did not affect the recovery time. The understanding of injury and recovery of E. coli could contribute to develop effective decontaminating treatment by lactic acid, and develop techniques for detecting sublethally injured E. coli cells.     

Antimicrobial efficacy of grape seed extract against Escherichia coli O157:H7 growth,motility and Shiga toxin production

Escherichia coli O157:H7 produces Shiga toxin (Stx) which is heat stable and causes Hemolytic Uremic Syndrome (HUS), a serious disease associated with bloody diarrhea and even death. To ensure food safety, both live E. coli O157:H7 and its toxin production in food products need to be controlled. Natural ingredients with inhibitory effects on E. coli O157:H7 growth and toxin production are top choices of antimicrobials for the food industry. The objectives of this study were to evaluate efficacy of grape seed extract (GSE) against the growth, swimming motility and Stx production of E. coli O157:H7. The disc diffusion assay indicated that 3.2 mg GSE per disc resulted in an inhibition zone of 14.8 ± 0.21 mm. The minimal inhibitory concentration of GSE against E. coli O157: H7 was 4.0 mg/ml. At high inoculation level (1 × 10⁷ CFU/ml), including GSE at 0.25–2.0 mg/ml reduced Stx production without inhibiting E. coli O157:H7 growth. At 5 × 10⁵ CFU/ml inoculation level, 2.0 and 4.0 mg/ml GSE effectively inhibited the growth of E. coli O157:H7 for at least 72 h, however, a low level of GSE (0.125–1.0 mg/ml) enhanced E. coli O157:H7 growth and Stx2 production. At 4 mg/ml, GSE completely abolished Stx2 production in addition to it bactericidal effect against E. coli O157:H7. In addition, GSE at concentration as low as 0.125% blocked the swimming motility, which is important for E. coli O157:H7 surface adherence. In conclusion, GSE is effective in inhibiting the motility of E. coli O157:H7, GSE shows potential to be used as a natural antimicrobial to control E. coli O157:H7.     

Effects of processing and storage variables on penetration and survival of Escherichia coli O157:H7 in fresh-cut packaged carrots

The effects of severity of slicing and peeling, and of storage atmosphere and temperature on the survival of Escherichia coli O157:H7 at and below the cut surfaces of fresh-cut carrot discs were determined. Slicing with a blunt machine blade enhanced penetration of E. coli O157:H7 and its subsequent survival during storage at 8 °C. Significantly (P < 0.05) higher numbers of cells (0.5 log CFU g⁻¹) penetrated deeper (475–500, 725–750 and 975–1000 μm) into carrot tissue sliced with a blunt machine blade compared to those sliced with a razor blade. Counts on Day 3 and Day 5 of storage remained higher in carrot tissue sliced with a blunt machine blade at all depths sampled. There were no significant effects of peeling method on penetration and survival. Storage of carrot slices in sub-optimal atmospheres (20%CO₂/1%O₂ resulted in increased survival of cells at the surface and within the tissue compared to storage in more optimal atmospheres (5% CO₂/3% O₂). Increasing storage temperature from 4 to 10 °C resulted in growth of E. coli O157:H7 at all sample depths. For all experiments, E. coli O157:H7 cells colonising the surface generally survived better than cells that penetrated into the tissue. The data are relevant to improving microbial safety in the fresh-cut sector by demonstrating that cutting with sharp blades (e.g. during harvesting and processing) reduces the depth of potential contamination by E. coli O157:H7. This may also have implications for more efficient anti-microbial dipping and reduced pathogen survival during storage.