Behaviour of Escherichia coli O157:H7 during the fermentation of Datta and Awaze, traditional Ethiopian fermented condiments, and during product storage at ambient and refrigeration temperatures

The survival of E. coli O157:H7 in fermenting foods and its prolonged survival in refrigerated fermented foods is documented. This prompted the study to evaluate survival of E. coli O157:H7 during the fermentation of Datta and Awaze, traditional Ethiopian condiments. Datta was prepared by wet milling a variety of spices along with green or red chilli and fermenting it by lactic acid bacteria. Awaze is a slurry made of red pepper, garlic and ginger to which various other spices were added and fermented by lactic acid bacteria (LAB) and yeasts. The Datta or Awaze slurry was separately inoculated with three strains of E. coli O157:H7 and the fermentation was allowed to proceed at ambient (20–25°C) temperatures for 7 days. When fermenting Datta or Awaze was initially inoculated at low inoculum level (3 log cfu/g), the test strains were not recovered after 24 h of fermentation. At higher initial inoculum level (6 log cfu/g), however, counts of the test strains in Datta at day 7 were less by about 1.5 log unit than the initial inoculum level. In fermenting Awaze, all test strains were completely eliminated in 7 days. The pH of the fermenting green and red Datta was reduced from 5.2 to 4.4 and that of Awaze dropped from 4.9 to 3.8 during this time. In another experiment, the fermented products were separately inoculated with the E. coli O157:H7 test strains at levels of 6 log cfu/g and incubated at ambient and refrigeration (4°C) temperatures for 7 days. In fermented Datta, two of the three strains were not recovered by enrichment after 6 days of storage at ambient temperatures. In fermented Awaze, all strains were below countable levels at day 5, but could still be recovered by enrichment at day 7. At refrigeration storage, counts of the test strains in Datta and Awaze products were <3 log cfu/g at day 7. The inhibition of our E. coli O157:H7 test strains in Datta and Awaze may be due to the antimicrobial activity of spices and other metabolites produced by LAB which may be effective at low pH.     

Evaluation of the effects of ultraviolet light on bacterial contaminants inoculated into whole milk and colostrum,and on colostrum immunoglobulin G

Raw milk and colostrum can harbor dangerous microorganisms that can pose serious health risks for animals and humans. According to the USDA, more than 58% of calves in the United States are fed unpasteurized milk. The aim of this study was to evaluate the effect of UV light on reduction of bacteria in milk and colostrum, and on colostrum IgG. A pilot-scale UV light continuous (UVC) flow-through unit (45 J/cm²) was used to treat milk and colostrum. Colostrum and sterile whole milk were inoculated with Listeria innocua, Mycobacterium smegmatis, Salmonella serovar Typhimurium, Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, and Acinetobacter baumannii before being treated with UVC. During UVC treatment, samples were collected at 5 time points and bacteria were enumerated using selective media. The effect of UVC on IgG was evaluated using raw colostrum from a nearby dairy farm without the addition of bacteria. For each colostrum batch, samples were collected at several different time points and IgG was measured using ELISA. The UVC treatment of milk resulted in a significant final count (log cfu/mL) reduction of Listeria monocytogenes (3.2 ± 0.3 log cfu/mL reduction), Salmonella spp. (3.7 ± 0.2 log cfu/mL reduction), Escherichia coli (2.8 ± 0.2 log cfu/mL reduction), Staph. aureus (3.4 ± 0.3 log cfu/mL reduction), Streptococcus spp. (3.4 ± 0.4 log cfu/mL reduction), and A. baumannii (2.8 ± 0.2 log cfu/mL reduction). The UVC treatment of milk did not result in a significant final count (log cfu/mL) reduction for M. smegmatis (1.8 ± 0.5 log cfu/mL reduction). The UVC treatment of colostrum was significantly associated with a final reduction of bacterial count (log cfu/mL) of Listeria spp. (1.4 ± 0.3 log cfu/mL reduction), Salmonella spp. (1.0 ± 0.2 log cfu/mL reduction), and Acinetobacter spp. (1.1 ± 0.3 log cfu/mL reduction), but not of E. coli (0.5 ± 0.3 log cfu/mL reduction), Strep. agalactiae (0.8 ± 0.2 log cfu/mL reduction), and Staph. aureus (0.4 ± 0.2 log cfu/mL reduction). The UVC treatment of colostrum significantly decreased the IgG concentration, with an observed final mean IgG reduction of approximately 50%. Development of new methods to reduce bacterial contaminants in colostrum must take into consideration the barriers imposed by its opacity and organic components, and account for the incidental damage to IgG caused by manipulating colostrum.     

Real-time polymerase chain reaction for the quantitative detection of tetA and tetB bacterial tetracycline resistance genes in food

A new, rapid, sensitive and specific method was developed to directly detect and quantify tetA and tetB in food. Both tet genes are two of the most frequently present tetracycline resistance genes in Gram-negative bacteria. A set of primers and Taqman probes was designed for each gene. The standard curves were performed using Escherichia coli BM13 (C600 RifR)/RP4 and E. coli NCTC 50365, which carry tetA and tetB, respectively. Meat and fish samples inoculated with these reference strains were used as a matrix to construct the standard curves for the analysis of 20 samples of chicken meat and 10 samples of hake (Merlucius merlucius). The limits of detection in pure culture were 5 cfu/mL (0.7 log cfu/mL) in the case of tetA, 50 cfu/mL (1.7 log cfu/mL) for tetB and 5 × 10² cfu/g (2.7 log cfu/g) for both genes in food samples. The results obtained by real-time quantitative polymerase chain reaction (qPCR) were compared to counts of tetracycline-resistant bacteria obtained by plating extracts of poultry and hake samples in culture media supplemented with 16 mg/L of tetracycline. Counts of tetracycline-resistant bacteria obtained by qPCR showed a positive correlation, especially interesting when compared with microbiological counts of tetracycline-resistant Enterobacteriaceae in poultry meat (r = 0.5509) and with tetracycline-resistant mesophilic aerobic bacteria in hake samples (r = 0.7146). The obtained results demonstrate that this method could be a useful tool for the direct quantification of the amount of bacterial strains that carry tetA and/or tetB genes in food samples.     

Identification of critical control points for control of microbiological contamination in processes leading to the production of ground beef at a packing plant

Carcasses, carcass portions and pieces of manufacturing beef were sampled by swabbing a 100 cm² randomly selected site on each of 25 randomly selected product units at each of several stages of the meat production processes at a large beef packing plant. After skinning, aerobes were recovered from each sample, at log mean numbers of 1.86 log cfu cm⁻²; and coliforms and Escherichia coli were recovered from <15 of 25 samples at log total numbers of 3.27 and 3.16 log cfu 2500 cm⁻². The numbers of bacteria recovered after preevisceration washing and spraying with 2% lactic acid solution were similar to the numbers recovered after skinning, and the numbers recovered from carcasses that had been eviscerated, split, vacuum/hot-water cleaned and trimmed were little different. However, the numbers of coliforms and E. coli were reduced by the washing of carcass sides; and pasteurizing followed by spraying with lactic acid reduced the numbers of aerobes to log mean numbers of 0.71 log cfu cm⁻² and coliforms and E. coli to log total numbers of 1.28 and 0.30 log cfu 2500 cm⁻², respectively. After cooling for 24 h, numbers of aerobes on carcass sides had increased to log mean numbers of 2.57 log cfu cm⁻², and log total numbers of coliforms and E. coli had increased to 3.34 and 2.18 log cfu 2500 cm⁻², respectively. Numbers had not increased after sides had been held for a further 12 h before breaking. Numbers of bacteria on forequarters increased during breaking operations on hanging sides and increased further after forequarters were dropped to conveyor belts. Numbers on hindquarters were little affected by those operations. Large numbers of bacteria were recovered from cleaned equipment that contacted forequarters. The numbers of bacteria on trimmings did not increase during conveying from breaking lines to bulk containers. The numbers of bacteria on manufacturing beef were less than the numbers on forequarters trimmings, but more than the numbers on hindquarters trimmings. The log mean numbers of bacteria in ground beef produced at the plant were 3.11 log cfu g⁻¹, and the log total numbers of coliforms and E. coli were 1.84 and 1.30 log cfu 25 g⁻¹, respectively. Similar numbers of bacteria were recovered from ground samples of manufacturing beef. It therefore appears that most of the E. coli as well as other bacteria in ground beef are deposited on the product during the initial operations for breaking forequarters. Thus, improvement of equipment cleaning rather than of production processes would be required to improve the microbiological condition of ground beef prepared at the plant. However, skinning, washing of dressed sides, pasteurizing and cooling are evidently critical control points in carcass production processes.     

Comparison of the Microbiological Quality of Environmentally Friendly and Conventionally Grown Vegetables Sold at Retail Markets in Korea

Fresh produce is usually eaten raw without cooking or heating, which may increase the probability of foodborne infection. The microbiological quality of 11 types of fresh, raw vegetables (romaine lettuce, sesame leaves, crown daisy, garlic chives, iceberg lettuce, cabbage, broccoli, leek, chili pepper, capsicum, and zucchini) purchased at retail markets in Iksan, Korea as affected by cultivation method (environmentally friendly vegetables [organic, pesticide‐free, and low‐pesticide vegetables] and conventionally grown vegetables) and harvest season was determined. Escherichia coli O157:H7 and Salmonella were not detected in all samples of vegetables tested. Aerobic mesophiles (>6 log cfu/g) were detected in environmentally friendly romaine lettuce and crown daisy and environmentally friendly and conventionally grown garlic chives, which also contained coliforms (>3 log cfu/g). Sesame leaf and crown daisy (regardless of cultivation method), as well as conventionally grown romaine lettuce and leek, contained >1 log cfu/g of E. coli. The overall microbiological quality of environmentally friendly and conventionally grown vegetables was not significantly different (P > 0.05). However, there were seasonal effects on populations of coliforms and generic E. coli on vegetables. The greatest numbers of microorganisms were isolated from environmentally friendly or conventionally grown vegetables purchased in winter. The vegetables, regardless of cultivation method or season, should be subjected to appropriate antimicrobial treatment to enhance their microbial safety.     

Changes of microbiological quality in meatballs after heat treatment

The effects of different cooking processes (grilling, oven and microwave cooking) on the microbial flora of the raw meatballs inoculated with E. coli O157:H7 at the level 2×10⁴ cfu/ml were investigated. The meatballs were stored at 4 °C. The flora of the raw meatballs is described in this paper. While Salmonella was found in each sample, none of the samples contained C. perfringens or E. coli O157:H7. The processes of grill cooking or microwave cooking decreased the microbial flora by 2–3 log cycles. E. coli O157:H7 was completely destroyed by all cooking methods. E. coli O157:H7 count of the raw meatball samples increased for 1.5 log cycles at the end of storage compared to beginning.     

Control of Escherichia coli O157:H7 in corn silage with or without various inoculants: Efficacy and mode of action

This study aimed to evaluate the effectiveness of 3 commercial bacterial inoculants at controlling Escherichia coli O157:H7 in corn silages during ensiling and feedout phases of silage production. A second objective was to determine whether the inoculants exhibited and transferred antibacterial activity against E. coli O157:H7 to the silages. Chopped corn forage was ensiled after treatment with the following: distilled water (control); 5 × 10⁵ cfu/g of E. coli O157:H7 (EC); EC and 1 × 10⁶ cfu/g of Pediococcus pentosaceus and Propionibacterium freudenreichii (EC+BII); EC and 1 × 10⁶ cfu/g of Lactobacillus buchneri (EC+LB); and EC and 1 × 10⁶ cfu/g of L. buchneri and P. pentosaceus (EC+B500). Each treatment was ensiled in triplicate in mini silos for 3, 7, 31, and 82 d and analyzed for pH and E. coli O157:H7 counts. Samples from d 82 were also analyzed for volatile fatty acids, lactate, and aerobic stability. Antibacterial activity of inoculants and silages was determined by the Kirby-Bauer disc diffusion test. The pH of silages from all treatments decreased below 4 within 3 d of ensiling and remained low until d 82. Therefore, E. coli O157:H7 was not detected in silages after any of the ensiling durations. Applying inoculants containing L. buchneri resulted in less lactate, more acetate, and greater aerobic stability compared with the control. Applying EC+BII containing P. freudenreichii did not increase propionate or aerobic stability. Subsamples of d 82 silages were reinoculated with 1 × 10⁵ cfu/g of E. coli O157:H7 either immediately after silo opening on d 82 or after 144 h of aerobic exposure (d 88), and E. coli were enumerated 24 h later. All silages reinoculated with the pathogen on d 82 had similar, low pH values (<4) and no E. coli were detected 24 h later. Control, EC, and EC+BII silages reinoculated with the pathogen after 144 h of aerobic exposure had relatively greater pH values (4.71, 5.67, and 6.03, respectively) and E. coli counts (2.87, 6.73, and 6.87 log cfu/g, respectively) 24 h later, whereas those treated with L. buchneri had low pH values (<4) and undetectable (EC+B500) or 10-fold lower (1.97, cfu/g; EC+LB) E. coli counts. All pure cultures of commercial bacterial inoculants exhibited antibacterial activity independent of pH against E. coli O157:H7, but the pH-independent activity did not persist in the treated silages, suggesting that E. coli elimination from silages was mediated by pH reduction.     

Impact of voltage and pulse delivery mode on the efficacy of pulsed light for the inactivation of Listeria

Listeria innocua inactivation by pulsed light (PL) was evaluated at different settings and voltages, to establish the best treatment conditions and post-treatment handling for further implementation of PL in the food industry. Fluences up to 0.2 J/cm² were applied to superficially inoculated TSA agar plates (4.5–5 log cfu/cm²). Inactivation was calculated, and log-linear and Weibull models were applied. A fluence of 0.2 J/cm² applied in a single pulse inactivated 3.8 log cfu/cm², while sequential application of this fluence yielded an inactivation between 1.5 and 2.5 log cfu/cm² depending on the delivery mode (consecutive flashing or with 5 min-holding times under ambient light or in the dark). Data from consecutive PL treatment were fitted with the Weibull model. No photoreactivation following PL was observed after 120-min exposure to ambient light in any of the conditions assayed. This study showed that flashing with a single pulse at higher voltage would offer the highest inactivation of Listeria.Industrial relevanceThis study offered information of practical interest to establish pulsed light processing and post-processing conditions for the control of Listeria spp. in the food industry, for instance in ready-to-eat (RTE) products. The use of higher voltages provided higher inactivation and allowed to minimise the number of flashes. If sequential treatments are to be applied, the treatment is more effective if short holding times are kept between pulses. The post-processing illumination conditions do not influence the efficacy of PL treatment.     

Influence of organic acid concentration on survival of Listeria monocytogenes and Escherichia coli 0157:H7 in beef carcass wash water and on model equipment surfaces

Acid-adapted cultures of Escherichia coli O157:H7 and Listeria monocytogenes were inoculated in meat decontamination spray-washing runoff fluids in order to evaluate their survival and potential to form biofilms on stainless steel coupons. The cultures (10⁷ cfu ml⁻¹) and stainless steel coupons were exposed to mixtures of water and organic acid washings (composites of each of 2% acetic acid or lactic acid washings with water washings from meat decontamination in proportions of 1/9, 1/49, 1/99 [vol/vol]) or to water washings for up to 14 days at 15°C. E. coli O157:H7 formed biofilms and remained detectable (1.3 log cfu cm⁻²) on stainless steel for up to 4 d in the 1/9 dilution (pH 3.17–3.77) of the organic acid washings, and persisted throughout storage (14 d) in the 1/49 (pH 3.96–4.33) and 1/99 (pH 4.34–6.86) dilution of the organic acid washings. L. monocytogenes populations were unable to form detectable (<1.3 log cfu cm⁻²) biofilms in the 1/9 and 1/49 dilutions of both organic acid washings for up to 14 d; however, by day-14 in the 1/99 dilution of the washings, the pathogen was able to attach at detectable levels (2.7 to 3.4 logs). The pH effects of lower concentrations (1/49 or 1/99) of acidic washings decreased over time due to the formation of amine compounds produced by the natural meat flora, allowing resuscitation of the acid-stressed pathogen survivors. The resuscitation of acid-stressed pathogens may potentially enhance their survival and prevalence in biofilms and thus more attention should be focused on avoiding or minimizing the collection of decontamination runoff fluids on food contact equipment surfaces.     

Impact of inulin and okara on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product and probiotic survival under in vitro simulated gastrointestinal conditions

The effect of inulin and/or okara flour on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product (FSP) and on probiotic survival under in vitro simulated gastrointestinal conditions were investigated throughout 28 days of storage at 4 °C. Employing a 2² design, four FSP trials were produced from soymilk fermented with ABT-4 culture (La-5, Bb-12, and Streptococcus thermophilus): FSP (control); FSP-I (with inulin, 3 g/100 mL of soymilk); FSP-O (with okara, 5 g/100 mL); FSP-IO (with inulin + okara, ratio 3:5 g/100 mL). Probiotic viabilities ranged from 8 to 9 log cfu/g during the 28 days of storage, and inulin and/or okara flour did not affect the viability of La-5 and Bb-12. Bb-12 resistance to the artificial gastrointestinal juices was higher than for La-5, since the Bb-12 and La-5 populations decreased approximately 0.6 log cfu/g and 3.8 log cfu/g, respectively, throughout storage period. Even though the protective effect of inulin and/or okara flour on probiotic microorganisms was not significant, when compared to a fresh culture, the FSP matrix improved Bb-12 survival on day 1 of storage and may be considered a good vehicle for Bb-12 and could play an important role in probiotic protection against gastrointestinal juices.     

Growth and survival of Escherichia coli O157:H7 during the fermentation and storage of diluted cultured milk drink

The fate of various Escherichia coli O157:H7 strains including 933, A8993-C32, MF6707, 18731A, EK250 and EF304 during the fermentation and storage of diluted cultured milk drink fermented with Lactobacillus casei ssp.casei CCRC 11197 or L. delbrueckii ssp. bulgaricus CCRC 14009 were investigated in this study. E. coli O157:H7, regardless of strains, grew rapidly in skim milk and reached a maximum population of c. 8·0–9·0 log cfu ml⁻¹after c. 24 h of cultivation in mixed cultures with L. casei ssp. casei or L. delbrueckii ssp. bulgaricus. However, the population of E. coli O157:H7, depending on the strain and the lactic acid bacteria present, declined as cultivation proceeded further. With the inoculation of c. 5·0 log cfu ml⁻¹E. coli O157:H7, viable cells of this pathogen reduced to non-detectable level in the non-sugar-added cultured drink (pH 3·5) prepared with L. delbrueckii ssp.bulgaricus after one day storage at 7°C. Depending on the strains, E. coli O157:H7 survived in the non-sugar-added cultured drink prepared with L. casei ssp. casei for a period of <1–4 days. Adding sugar to cultured drink extended the survival period of E. coli O157:H7. The extent of the sugar-protective effect varied with different strains of E. coli O157:H7 and the amount of sugar added to the drink.     

The effect of different cooking procedures on microbiological and chemical quality characteristics of Tekirdaǧ meatballs

In this research, the effects of different cooking processes (grilling, oven, and microwave cooking) on microbial flora and chemical composition of the raw and cooked meatballs as consumed in Tekirdaǧ were investigated. Microbial flora of the raw meatballs was as follows: total bacteria, 6.02×10⁶ cfu/g; psychrophilic bacteria, 1.3×10⁵ cfu/g; yeast and mould, 2.4×10⁵ cfu/g; coliforms, 1.1×10⁵ cfu/g; Escherichia coli, 1.0×10² cfu/g; total staphylococcae, 3.3×10² cfu/g; Staphylococcus aureus, 85 cfu/g. While Salmonella was found in only one sample, none of the samples contained Clostridium perfringens. The cooking processes clearly decreased the microbial flora (2–3 log cycles in grilling (71°C) and oven‐cooked (79°C( 3–4 log cycles in microwave (97°C) heating) of the meatballs. However, because of the crust formation and high moisture losses from the meatball surface in microwave heating, some sensorial defects were observed in the final product. Also, fat and moisture losses were higher in microwave cooking compared to the other cooking processes. In conclusion, it is advised to use slightly higher temperatures than used in the grilling or conventinal cooking procedures to increase microbial quality of the meatballs studied in this research.     

Reduction of Escherichia coli O157:H7 in Biofilms Using Bacteriophage BPECO 19

Biofilm formation is a growing concern in the food industry. Escherichia coli O157:H7 is one of the most important foodborne pathogens that can persists in food and food‐related environments and subsequently produce biofilms. The efficacy of bacteriophage BPECO 19 was evaluated against three E. coli O157:H7 strains in biofilms. Biofilms of the three E. coli O157:H7 strains were grown on abiotic (stainless steel, rubber, and minimum biofilm eradication concentration [MBEC^TM] device) and biotic (lettuce) surfaces at different temperatures. The effectiveness of bacteriophage BPECO 19 in reducing preformed biofilms on these surfaces was further evaluated by treating the surfaces with a phage suspension (10⁸ PFU/mL) for 2 h. The results indicated that the phage treatment significantly reduced (P  < 0.05) the number of adhered cells in all the surfaces. Following phage treatment, the viability of adhered cells was reduced by ≥3 log CFU/cm², 2.4 log CFU/cm², and 3.1 log CFU/peg in biofilms grown on stainless steel, rubber, and the MBEC^TM device, respectively. Likewise, the phage treatment reduced cell viability by ≥2 log CFU/cm² in biofilms grown on lettuce. Overall, these results suggested that bacteriophages such as BPECO 19 could be effective in reducing the viability of biofilm‐adhered cells.     

UV-C inactivation of Escherichia coli at different temperatures

The influence of treatment parameters (dose and temperature), treatment medium characteristics (absorption coefficient, pH and water activity) and microbiological factors (strain, growth phase and UV damage and repair capacity) on Escherichia coli UV-C resistance has been investigated. UV-C doses to inactivate at 25 °C 99.99% of the initial population (4D) of five strains of E. coli in McIlvaine buffer of pH 7.0 with tartrazine added (absorption coefficient of 10.77 cm⁻¹) were 16.60, 14.36, 14.36, 13.22, 11.18 J/mL for strains E. coli STCC 4201, STCC 471, STCC 27325, O157:H7 and ATCC 25922, respectively. The entrance in the stationary growth phase increased the 4D value of the most resistant strain, E. coli STCC 4201, from 13.09 to 17.23 J/mL. Survivors to UV treatments showed neither oxidative damages nor injuries in cell envelopes. On the contrary, the photoreactivation by the incubation of plates for 60 min below visible light (11.15 klx) increased the dose to 18.97 J/mL. The pH and the water activity of the treatment medium did not affect the UV tolerance of E. coli STCC 4201, but the lethal effect of the treatments decreased exponentially (Log₁₀4D = − 0.0628α + 0.624) by increasing the absorption coefficient (α). A treatment of 16.94 J/mL reached 6.35, 4.35, 2.64, 1.93, 1.63, 1.20, 1.02 and 0.74 Log₁₀ cycles of inactivation with absorption coefficients of 8.56, 10.77, 12.88, 14.80, 17.12, 18.51, 20.81 and 22.28 cm⁻¹. The temperature barely changed the UV resistance up to 50.0 °C. Above this threshold, inactivation rates due to the combined process synergistically increased with the temperature. The magnitude of the synergism decreased over 57.5 °C. An UV treatment of 16.94 J/mL in media with an absorption coefficient of 22.28 cm⁻¹ reached 1.23, 1.64, 2.36, 4.01 and 6.22 Log₁₀ cycles of inactivation of E. coli STCC 4201 at 50.0, 52.5, 55.5, 57.5 and 60.0 °C, respectively.

Industrial relevance

Results obtained in this investigation show that UV light applied at mild temperatures (57.5 to 60 °C) could be an alternative to heat treatments for 5-Log₁₀ reductions of E. coli in liquid foods. Since microbial resistance to UV-C light did not depend on the pH and water activity (a_w) of the treatment media, eventual advantages of UV light for pasteurization purposes will be higher in low a_w foods. E. coli STCC 4201 could be considered as a target when UV light processing of foods.     

Microbiological Quality and Safety of Fresh Fruits and Vegetables at Retail Levels in Korea

The aim of this study was to evaluate the microbiological quality and safety of fresh produce at retail level in Korea in order to periodically update information and establish available risks associated with consumption of fresh fruits and vegetables. The samples from different markets located in 3 provinces of South Korea were collected. The protocol in the Korean Food Standards Codex was applied and generic Escherichia coli, coliforms, aerobic mesophilic bacteria (AMB), and yeast and mold (YM) in 360 packaged and unpackaged fresh fruits and vegetables were analyzed. Presence of pathogens was examined using real‐time polymerase chain reaction (q‐PCR) after enrichment of samples. For all, the microbial counts ranged from 1.7 to 10.6 log cfu/g for AMB, 2.2 to 7.9 log cfu/g for coliforms, and 5.5 to 7.9 log cfu/g for YM. Three lettuce samples were contaminated by E. coli with a bacterial load ranging from 2 to 4 log cfu/g. Salmonella spp. were not detected in any fresh produce. Listeria monocytogenes, E. coli O157:H7, and Staphylococcus aureus were found in 1 (0.6%), 3 (0.8%), and 5 (1.4%) fresh produce samples, respectively. Bacillus cereus (50.3%) and Clostridium perfringens (13.3%) had the highest prevalence. These results indicate the need for employing strict control measures and developing preventive strategies to improve the quality and safety of fresh produce in Korea.     

Efficacy of heat treatment in the reduction of salmonellae and Escherichia coli O157:H– on alfalfa, mung bean and radish seeds used for sprout production

We studied the effect of hot-water treatment at various time/temperature regimes to design a decontamination process which is consistent with the recommendation of the National Advisory Committee on Microbiological Criteria for Foods (NACMCF) to reduce pathogens on seeds by 5log cfu/g. Alfalfa, mung bean and radish seeds were inoculated by immersion with more than 10⁷ cfu/g of enterobacteria (Salmonella Senftenberg W775, S. Bovismorbificans and Escherichia coli O157:H^–), dried and stored at 2 °C. The numbers of salmonellae and E. coli O157:H^– on these seeds remained unchanged during storage for 8 weeks. To achieve sprouting rates of more than 95%, time-temperature regimes were defined. The thermal treatment of contaminated mung bean (2–20 min for 55–80 °C), radish and alfalfa seeds 0.5–8 min (53–64 °C) reduced all pathogens by more than 5log cfu/g. For S. Senftenberg W775 on radish seeds, D values of 3.2, 1.9 and 0.6 min were determined for exposure at 53, 55 and 58 °C and a z value of 6.2 °C was calculated. For alfalfa seeds, the respective D values were 3.0, 1.6, and 0.4 min and the z value was the same as that determined for radish seeds.     

Ultraviolet and Pulsed Electric Field Treatments Have Additive Effect on Inactivation of E. coli in Apple Juice

ABSTRACT: Apple juice inoculated with Escherichia coli ATCC 23472 was processed continuously using either ultraviolet (UV), high‐voltage pulsed electric field (PEF), or a combination of the PEF and UV treatment systems. Apple juice was pumped through either of the systems at 3 flow rates (8, 14, and 20 mL/min). E. coli was reduced by 3.46 log CFU/mL when exposed in a 50 cm length of UV treatment chamber at 8 mL/min (2.94 s treatment time with a product temperature increase of 13 °C). E. coli inactivation of 4.87 log CFU/mL was achieved with a peak electric field strength of 60 kV/cm and 11.3 pulses (average pulse width of 3.5 μs, product temperature increased to 52 °C). E. coli reductions resulting from a combination treatment of UV and PEF applied sequentially were evaluated. A maximum E. coli reduction of 5.35 log CFU/mL was achieved using PEF (electrical field strength of 60 kV/cm, specific energy of 162 J/mL, and 11.3 pulses) and UV treatments (length of 50 cm, treatment time of 2.94 s, and flow rate of 8 mL/min). An additive effect was observed for the combination treatments (PEF and UV), regardless of the order of treatment (P > 0.05). E. coli reductions of 5.35 and 5.30 log CFU/mL with PEF treatment (electrical field strength of 60 kV/cm, specific energy of 162 J/mL, and 11.3 pulses) followed by UV (length of 30 cm, treatment time of 1.8 s, and flow rate of 8 mL/min) and UV treatment followed by PEF (same treatment conditions), respectively. No synergistic effect was observed.     

An innovative water-assisted UV-C disinfection system to improve the safety of strawberries frozen under cryogenic conditions

Strawberries inoculated with Salmonella enterica, Listeria monocytogenes (10⁸ CFU/mL, 50 μL) and murine norovirus (MNV-1; 10⁶ TCID₅₀/mL, 50 μL), were washed for 2 min in a water-assisted UV-C light tank (WUVC) combined or not with 40 mg/L of peracetic acid (WUVC+PA), and 200 mg/L of free chlorine solution (NaClO) with the UV-C lamps switched off. Moreover, a ‘conventional’ dry UV-C treatment (DUVC) was also tested. After 2-min exposure, washing sanitization with chemical agents gave the highest reduction for both bacteria (ca. ≥ 3.3 log CFU/g) and MNV-1 (≥1.8 log TCID₅₀/mL). DUVC treatment proved to be the least effective technology (≤0.6 log CFU/g for bacteria and 1.5 log TCID₅₀/mL for MNV-1). Regarding wash water, no presence of L. monocytogenes and S. enterica were reported with WUVC+PA and NaClO sanitization. After disinfection, samples were frozen at −70 ± 2 °C in a cryogenic freezing cabinet with liquid nitrogen (N₂). For both pathogens, frozen storage after washing substantially enhanced their inactivation in the first 3 days (1.1–4.9 log UFC/g) compared to the reductions obtained the following sampling points (0.0–0.8 log UFC/g). After 90 days, L. monocytogenes and S. enterica were not detected on the samples treated with water-assisted methodologies (WUVC, WUVC+PA and NaClO treatments), whilst MNV-1 was little affected. Further studies are needed to improve norovirus inactivation on frozen strawberries.Industrial relevanceThe present work provides relevant information to the frozen food industry regarding a suitable decontamination alternative to chlorine sanitation. Low-dose immersion-assisted UV-C allows inactivation and inhibition of pathogenic microbiota while generates non-toxic byproducts and allows reusing the process water, contributing to the so-called “smart green growth” attended to provide a more innovative and sustainable future for the food industry.     

Repair and growth of heat- and freeze-injured Escherichia coli O157:H7 in selective enrichment broths

Two Escherichia coli O157:H7 strains, ATCC 35150 and 43894, were heat injured in a beef infusion at 53°C for 40 and 50 min, respectively (1· 5–2·0 log₁₀cfu ml⁻¹of injury) and freeze injured at −25°C for 30 days (1 log₁₀cfu ml⁻¹of injury) as determined by plating on MacConkey agar with 0·60% bile salts #3 (Mac-BS) as the selective medium and on Brain Heart Infusion agar (BHIA) as the non-selective medium. Repair of injury was measured in five selective enrichment broths [buffered peptone water supplemented with vancomycin, cefsulodin, and cefixime (BPW-VCC), modified EC broth with novobiocin (mEC+n), enterohaemorrhagic E. coli enrichment broth (EEB), double modified TSB (dmTSB), and BCM^®E. coli enrichment broth (BCM^®-EB)] versus TSB as the non-selective control broth over 3 h incubation at 37°C and 42°C. Repair was measured as the increase in cfu ml⁻¹enumerated on Mac-BS with time vs the total cfu ml⁻¹(injured and uninjured cells) enumerated on BHIA. In mEC+n, EEB, and dmTSB some death of both heat- and freeze-injured cells occurred immediately during the 3 h incubation (decrease on BHIA plates), and there was either minimal or no repair of the injured cells at both temperatures. Efficient repair of heat injury was obtained with both BPW-VCC and BCM^®-EB, but the latter produced a growth rate and final cell concentration closer to TSB. In freeze-injury repair however, BPW-VCC gave poor results while repair in BCM^®-EB was equal to TSB. Both BCM^®-EB and BPW-VCC inhibited the growth of all Gram-positive and a select number of Gram-negative bacteria tested. The ability of the selective enrichment broth BCM^®-EB to resuscitate heat- and freeze-injured E. coli O157:H7 efficiently within 3 h, warrants further testing with other types of stress in both artificially and naturally contaminated foods.     

Thermal inactivation of Escherichia coli O157:H7 and Escherichia coli isolated from morcilla as affected by composition of the product

Morcilla is a link sausage quite similar to black pudding, consisting of an inert casing stuffed with a mixture of beef blood, fat, and seasonings. Thirty samples of morcilla showed total microbial counts (6.3×10³–2.1×10⁸ Cfu/g ), molds and yeasts (8.9×10¹–6.3×10⁴ Cfu/g), sulfite-reducing microorganism (2.0×10¹–2.1×10²MPN/g); total coliforms (1.4×10¹–1.1×10³ MPN/g); fecal coliforms (7.0–1.5×10²MPN/g); Enterobactereaceae (1.6×10²–5.0×10⁵ Cfu/g). S. aureus and B. cereus were not detected. E. coli was detected in 76.6% of the samples analyzed. The thermal resistance (D and z-values) of Escherichia coli O157:H7 and E. coli isolated from morcilla were determined in nutrient broth and in a heating menstruun prepared with ground morcilla (discarding the casing) and added fat or starch. Higher fat and starch levels resulted in higher D-values (min) at 54, 58 and 62 °C for both strains. The z-values (°C) for isolated E. coli in nutrient broth (M1), ground morcilla (M2), M2+10% fat (M3), M2+20% fat (M4), M2+10% starch (M5), and M2+20% starch (M6) were 7.9, 7.8, 10.5, 10.4, 10.3, and 10.4, respectively, and for E. coli O157:H7 were 7.8, 7.4, 9.8, 10.2, 10.3, and 10.7. The composition of product affected heat lethality of the two strains of E. coli.