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Dietary supplementation of probiotics is growing as a scientifically valid alternative to antibiotics for enhancement of overall animal health and productivity in aquaculture. Strains of Bacillus subtilis are regarded as attractive probiotic candidates to the fish farming industry; however, there is a limited number of studies focused on the use of specific strains probiotics in tilapia, and therefore complicating replication. The objective of this study was to examine the effect of the strains NZ86 (NRRL B-50136) and O14VRQ (NRRL B-67221) of B. subtilis on various parameters of the innate immunity in Nile tilapia (Oreochromis niloticus) in a 51-day feeding trial. Supplementation of tilapia with either strain resulted in significant increases (p 0.05) in plasma lysozyme concentration of varying degrees throughout the trial. Meanwhile, alternative complement activity was significantly elevated (p 0.05) only after feeding of the NZ86 strain after 14 and 51 days. Conversely, supplementation with O14VRQ resulted in a significant increase (p 0.05) in the percent of neutrophils in the peripheral blood of tilapia by day 28. At the end of the trial, there was a trend towards increased phagocytic and respiratory burst activities observed in immune organ derived leukocytes. Feeding with either probiotic appeared to have an up-regulation on the gene expression of both pro-inflammatory cytokines in the intestine, yet only O14VRQ was significantly different than the control. Moreover, the occurrence of these results could be associated with supplementation of the probiotic strains, given that Bacillus bacteria were observed to populate the intestines of the dietary treatment groups. These results suggest the potential roles of these B. subtilis probiotic candidates to stimulate immune responses both locally and systemically in tilapia.

Salmonella serotypes linked to tomato-associated outbreaks were evaluated for survival in soil and water over a 40-day period. Salmonella enterica serotypes Anatum, Baildon, Braenderup, Montevideo, Newport, and Javiana were inoculated separately into sterile soil and water, followed by plating onto TSAYE and XLT4 at 10-day intervals. Biofilm production by Salmonella serotypes was measured on both quartz particles (soil surrogate) and glass coverslips, and was evaluated using a crystal violet dye assay. Salmonella populations in soil and water over 40 days indicated no significant differences between Salmonella serotypes tested (p > 0.05). Over a 40-day period, there was a 1.84 ± 0.22 log CFU/g and 1.56 ± 0.54 CFU/mL decrease in populations of Salmonella in soil and water, respectively. Enumeration indicated that Salmonella population fluctuated in water but decreased linearly in soil. All serotypes tested produced the "red dry and rough" morphotype on Congo Red agar. Biofilm produced by all the Salmonella serotypes tested was significantly different on quartz particles than on glass coverslips (p

The presence of dust is ubiquitous in the produce growing environment and its deposition on edible crops could occur. The potential of wind-distributed soil particulate to serve as a vehicle for S. Newport transfer to tomato blossoms and consequently, to fruits, was explored. Blossoms were challenged with previously autoclaved soil containing S. Newport (9.39log CFU/g) by brushing and airborne transfer. One hundred percent of blossoms brushed with S. Newport-contaminated soil tested positive for presence of the pathogen one week after contact (P

In May 2016, labeling of certain nonintact mechanically tenderized beef (MTB) products was mandated in the United States. MTB products should be handled differently by the consumer because pathogens can be transferred from the exterior to the interior of the meat during the tenderization process. Without labeling, it is difficult to visually distinguish between some intact beef and MTB products, which is a concern because MTB products require higher internal cooking temperatures for safety. An exploratory study was conducted to understand consumer understanding of MTB products and consumer responses to the new label. Thirteen focus groups were convened in rural and urban settings across Virginia and North Carolina between December 2015 and May 2016. Sessions were audiorecorded, transcribed verbatim, and analyzed through constant-comparison thematic analysis. Although MTB products were commonly bought, prepared, and consumed, consumer awareness of MTB products and the MTB process was limited. Generally, the label confused participants, and they did not understand the message. Specifically, terminology such as "blade tenderized" and "mechanically tenderized" were preferred over the term "needle tenderized" on labels. Once explained, many individuals wanted more information and better messaging. Through a multiprong approach, other messaging methods (e.g., in stores, through technology, and with certifications) were highly valued by consumers and may result in increased message clarity. Ultimately, the intrinsic and extrinsic properties of the beef rather than the MTB product continued to be the primary guide for purchasing and preparation. This study is the first to be conducted regarding American perceptions of MTB products. An understanding of consumer awareness of MTB products and labels is needed to develop targeted risk messaging communication tools.

Mechanically tenderized beef (MTB) products are non-intact beef products that have been tenderized using blades, needles, or other mechanical processes to improve consumer palatability. MTBs have been associated with outbreaks of Shiga toxin-producing Escherichia coli. MTB labeling was mandated in May 2016; however, consumer awareness of product and label is unclear. A national survey quantified consumer understanding and attitudes surrounding MTBs and its preparation. Many (64%) respondents were unaware that beef is mechanically tenderized; MTB label awareness was also low. Thirty-eight percent of respondents familiar with MTBs held positive attitudes towards MTBs due to price and convenience. Negative attitudes (24%) towards MTBs stemmed from concerns with the process, its “natural”-ness, and safety. Respondents regularly preparing MTBs reported using inside and/or outside color of the beef product (63%), texture (43%), juice/blood appearance (47%), smell (22%), and/or time (51%) to indicate doneness. While 20% of respondents used a thermometer, 62% reported “always” or “most of the time” preparing MTBs to 160℉. Contradictions may result from social desirability bias and optimism bias to portray selves in a positive light while believing personal preparation methods achieve safe results. Timely and targeted messaging with clear and standardized definitions, developed with consumers, are needed to bring awareness of MTBs and other food safety information to all populations.

The efficacy of ozone and ultraviolet light (UV) treatment as hurdles against Listeria monocytogenes suspended in fresh (9% NaCl, 91.86% transmittance) and spent brines (20.5% NaCl, 0.01% transmittance) was evaluated. Brines were inoculated with a cocktail of L. monocytogenes-strains N1-227, N3-031, and R2-499. Ozonation was performed by sparging gaseous ozone into brine. This was followed by UV irradiation (253.7 nm) of the brine in sterile quartz cuvettes. Enumeration was performed by spread plating on modified Oxford medium and Trypticase Soy agar supplemented with yeast extract. In fresh brines containing L. monocytogenes, 10 min of ozonation lead to a 7.44 ± 0.13 log CFU/ml mean reduction and 10 min of UV radiation caused a 1.95 ± 0.41 log CFU/ml mean reduction. Sequential exposure of 10 min of ozonation and UV resulted in >9 log CFU/ml reduction in L. monocytogenes populations in fresh brine. Sixty minutes of ozonation of spent brines resulted in a 4.85 ± 0.61 log CFU/ml mean reduction of L. monocytogenes populations. Ten minutes of UV exposure in spent brines resulted in 0.49 ± 0.14 log CFU/ml mean reduction in L. monocytogenes. A sequential treatment of 60 min ozonation and 10 min UV resulted in an excess of 5 log CFU/ml reduction in L. monocytogenes cells in spent brine. Ozonation did not cause a significant increase in the transmittance of the spent brine to aid UV penetration but resulted in color change. Sequential treatments of Ozonation and UV maybe effective in reducing L. monocytogenes in chill brines.

Salmonella, Shiga toxin-producing Escherichia coli (STEC), and Listeria monocytogenes have been isolated from low water activity foods (LWAF), where they may survive for extended periods. The ready-to-eat nature of many LWAF, such as dried fruits and nuts, warrants effective post-harvest thermal treatment for the reduction of pathogens such as low-temperature, saturated steam, also known as vacuum-assisted steam pasteurization. The objective of this study was to determine reductions of Salmonella, STEC, L. monocytogenes, and a possible surrogate (Pediococcus acidilactici) on dried apricot halves, whole macadamia nuts, and raisins after treatment with vacuum-assisted steam at three temperatures (62 °C, 72 °C, or 82 °C) and multiple time intervals. Bacterial inactivation was variable between commodities, with higher temperatures and longer times necessary to achieve comparable reductions of pathogens on apricot halves and macadamia nuts compared to raisins. Reductions of the tested pathogens were comparable; therefore, one species was not more resistant than the others. Pathogens were reduced by 5-log CFU/g on apricot halves after 20 min at 72 °C and after 5 min at 82 °C. Longer treatment times were necessary to achieve reductions of each pathogen on macadamia nuts. Pathogens were reduced by nearly 5 log CFU/g on macadamia nuts after 38 min at 72 °C (4.6–6.5 log CFU/g) and after 12 min at 82 °C (4.9–5.7 log CFU/g). Reductions of pathogens on raisins were achieved at lower temperatures than necessary for the other foods. A 5-log reduction for each of the pathogens (CFU/g) on raisins occurred after 20 min at 62 °C and after 5 min at 72 °C. Overall, the reductions of the pathogens exceeded those of P. acidilactici on both the dried fruits and macadamia nuts. Statistically significant differences, indicating greater confidence as a conservative surrogate, were observed at lower treatment temperatures. Inactivation kinetics were modeled for each pathogen on each food type and temperature. Bacterial survival was best described by the Weibull model for raisins and macadamia nuts, while the Gompertz model best described reductions on apricot halves according to Akaike information criterion (AIC) and root-mean-square error (RMSE) evaluations. Water activity and moisture content were increased due to the treatments, which could be addressed through implementation of drying steps. Thermal inactivation kinetic models and 5-log reduction parameters can help food processors design and evaluate similar vacuum-assisted steam interventions to comply with FSMA regulations and preventive control plans. However, results or model predictions should not be extrapolated to assume the safety of other types of foods.

Spices, including black pepper and cumin seeds, have been implicated in recent outbreaks of salmonellosis and prompted recalls of ready-to-eat products containing contaminated spices. Vacuum-assisted steam pasteurization is performed to improve the safety and quality of many low water activity products, however process parameters associated with inactivation on whole spices are not well described. The objective of this study was to determine the effectiveness of a lab-scale vacuum-assisted steam process for the inactivation of Salmonella enterica and its potential surrogate Enterococcus faecium ATCC 8459 inoculated onto the surface of whole peppercorns and cumin seeds. In addition, the effect of two inoculation preparation methods (growth on tryptic soy agar (TSA) or inclusion within a native microbiota biofilm), on the reduction of S. enterica serovars or E. faecium was compared on steam pasteurized whole black peppercorns. Spices were processed using steam under a vacuum to achieve a mean product temperature of 86.7 ± 2.8 °C for different dwell times. Salmonella inoculated using the TSA-grown method, required 83 s and 70 s respectively to achieve a 5-log reduction of Salmonella on peppercorns and cumin seeds. Longer time periods were needed to achieve a 5-log reduction of Salmonella when it was present in a native biofilm on whole peppercorns. Survivor estimations were best predicted by the Weibull models. The mean log reductions of E. faecium were 0.9 log CFU/g lower than Salmonella on whole black peppercorns inoculated using the TSA-grown cells (P = 0.0021). The mean log reductions of Salmonella and E. faecium prepared using the biofilm-inclusion method were not significantly different (P = 0.76). E. faecium log CFU/g reductions were not significantly different compared to Salmonella on whole cumin seeds (P = 0.42) indicating that while reductions are comparable the surrogate may not always provide a conservative indication of complete Salmonella elimination for all spices processed using vacuum-assisted steam.

Cetylpyridinium chloride (CPC) solutions (0, 0.5, or 1.0%) were applied to cantaloupe (Athena and Hale's Best Jumbo cultivars) rind plugs, either before or after inoculation with a broth culture of Salmonella Michigan (10(9) CFU/mL) and held at 37 degrees C for 1 or 24 hr. Rind plugs were diluted, shaken, and sonicated, and solutions were enumerated. Texture quality and color were evaluated over 14 days storage at 4 degrees C after 0 and 1% CPC spray applications. A 0.5 or 1.0% (vol/vol) application of CPC after Salmonella reduced the pathogen levels between 2.34 log CFU/mL and 5.16 log CFU/mL in comparison to the control (p

Propylene oxide (PPO), a chemical fumigant, has been validated to reduce Salmonella on bulk almonds but has not been evaluated for other tree nuts. There is a need to identify nonpathogenic surrogate microorganisms whose inactivation is comparable to that of Salmonella to assure effective PPO processing parameters in different packaging configurations without introducing Salmonella into the pasteurization facility. The objective of this research was to compare the reduction of Salmonella and three potential surrogate bacterial strains, Enterococcus faecium ATCC 8459, Pediococcus acidilactici ATCC 8042, or Staphylococcus carnosus ATCC 51365, on cashews and macadamia nuts processed by using PPO. Whole cashews and macadamia nuts were coinoculated with a five-strain cocktail of Salmonella and one surrogate, dried to the original water activity of 0.44 to 0.51 before being packaged in woven polypropylene bags (2.3 kg), and shipped overnight in Styrofoam containers under ambient conditions to a commercial facility for PPO treatment. Salmonella and surrogates were recovered by vigorous shaking in phosphate buffer (1:1, m/v), serial diluted, and plated onto tryptic soy agar with an overlay of xylose lysine Tergitol 4 for Salmonella or mannitol salt agar or bile esculin azide agar for each surrogate. The mean log reductions of Salmonella and each surrogate ( n = 18), within a sample and among all trials (three independent), were compared by using a matched pairs t test. Reduction in log CFU per gram of Salmonella was significantly greater than that of E. faecium on both macadamia nuts (7.3 ± 0.19 versus 6.4 ± 0.31) and cashews (5.4 ± 0.15 versus 5.1 ± 0.25) and significantly greater than P. acidilactici on both nuts (7.8 ± 0.22 versus 6.3 ± 0.33 on macadamia nuts and 4.9 ± 0.22 versus 4.1 ± 0.25 on cashews). Reduction of S. carnosus exceeded that of Salmonella. E. faecium and P. acidilactici may be considered as surrogates for Salmonella on whole macadamia nuts and cashews processed by using PPO.

Salmonellosis has been increasingly associated with contaminated spices. Identifying inoculation and stabilization methods for Salmonella on whole spices is important for development of validated inactivation processes. The objective of this study was to examine the effects of inoculation preparation on the recoverability of Salmonella enterica from dried whole peppercorns and cumin seeds. Whole black peppercorns and cumin seeds were inoculated with S. enterica using one dry transfer method and various wet inoculation methods: immersion of spice seeds in tryptic soy broth (TSB) plus Salmonella for 24 h (likely leading to inclusion of Salmonella in native microbiota biofilms formed around the seeds), application of cells grown in TSB, and/or application of cells scraped from tryptic soy agar (TSA). Postinoculation seeds were dried to a water activity of 0.3 within 24 h and held for 28 days. Seeds were sampled after drying (time 0) and periodically during the 28 days of storage. Salmonella cells were enumerated by serial dilution and plated onto xylose lysine Tergitol (XLT4) agar and TSA. Recovery of Salmonella was high after 28 days of storage but was dependent on inoculation method, with 4.05 to 6.22 and 3.75 to 8.38 log CFU/g recovered from peppercorns and cumin seeds, respectively, on XLT4 agar. The changes in surviving Salmonella (log CFU per gram) from initial inoculation levels after 28 days were significantly smaller for the biofilm inclusion method (z0.142pepper, z0.186cumin) than for the other inoculation methods (20.425pepper, 22.029cumin for cells grown on TSA; 20.641pepper, 20.718cumin for dry transfer; 21.998pepper for cells grown in TSB). In most cases, trends for reductions of total aerobic bacteria were similar to those of Salmonella. The inoculation method influenced the recoverability of Salmonella from whole peppercorns and cumin seeds after drying. The most stable inoculum strategies were dry transfer, 24-h incubation of Salmonella and spices in TSB (i.e., potential inclusion of Salmonella within native microbiota biofilms), and inoculation of Salmonella cells grown on TSA subsequent to drying. However, with the dry transfer method it was difficult to obtain the large amount of inoculum needed for inactivation studies. Published version This article was submitted using Open Access

Listeria monocytogenes is a foodborne pathogen that represents a high risk for consumers because it can grow under refrigeration conditions and can also develop acid tolerance. Fresh blueberries are hand-picked, packed, and transported under refrigeration without receiving a microbial inactivation treatment. The aim of this work was to study the survival of L. monocytogenes in fresh highbush blueberries stored at 4 or 12 °C under different controlled atmosphere conditions, including air (control); 5% O2, 15% CO2, 80% N2 (controlled atmosphere storage [CAS]); or ozone gas (O3), 4 ppm at 4 °C or 2.5 ppm at 12 °C, at high relative humidity (90 to 95%) for a total of 10 days. Fresh blueberries inside a plastic clamshell were spot inoculated with the bacteria and were stored at 4 or 12 °C in isolated cabinets under air, CAS, and O3 atmospheric conditions. Samples were evaluated on days 0, 1, 4, 7, and 10 for microbial growth using modified Oxford agar. CAS did not delay or inhibit L. monocytogenes growth in fresh blueberries after 10 days. O3 achieved 3- and 2-log reductions when compared with air treatment at 4 and 12 °C, respectively. Low concentrations of O3 together with proper refrigeration temperature can ensure product safety throughout transportation. O3 is a strong antimicrobial that safely decomposes to oxygen and water without leaving residues and can be used as an alternative method to prevent bacterial growth during a long transport period.

Postprocessing contamination of the products in a processing plant has been identified as one of the major reasons for food contamination with Listeria; brining is one such postprocessing area. Our previous study has shown that the combinations of UV and antimicrobials reduces the number of this organism significantly in fresh brine, but brine is usually recycled from days to weeks depending on its use. Therefore, this study is focused on the reduction of L. monocytogenes in recycled chill brine (obtained from a frankfurter processor) using the combinations of UV and antimicrobial agents, such as citric acid (CA), hydrogen peroxide (HP) and dimethyl dicarbonate (DMDC). Results show that the combinations of UV and 2000 ppm and 4000 ppm HP were the most effective treatments in reducing the Listeria population with the total processing time of 120 min. Both of these treatments were found to be more effective than UV or HP alone. Additionally, all other treatments, such as the combinations of UV and CA (0.2 and 0.5%) and UV and DMDC (250 and 500 ppm) were comparatively less effective. This may be due to the presence of organic matter in spent brine, which may have reduced the penetration of UV and availability of antimicrobials for microbial interaction. PRACTICAL APPLICATIONS The brine solutions are generally recycled in industries for up to 4 weeks as per United States Department of Agriculture (USDA) guide. Thus, it is very important to study various treatments to process recycled brine that was used to process ready-to-eat (RTE) meat products to prevent foodborne diseases or outbreaks. The use of UV light and antimicrobial agents to control L. monocytogenes in recycled brines to product cooling in the RTE meat industry is described. The study provides information on exposure time to UV light and level of antimicrobial agent that result in inactivation of the pathogen. Industry may benefit from our research in their validation efforts to control L. monocytogenes for recycled brine in recirculating chill brine systems through the use of UV light and antimicrobial agents.

Chill brine used during ready-to-eat meat processing is an important source of postprocessing contamination by Listeria monocytogenes. The efficacy of UV in combination with citric acid (CA; 0.2 and 0.5%), dimethyl dicarbonate (DMDC; 250 and 500 ppm) or hydrogen peroxide (2,000 and 4,000 ppm) was determined to reduce L. monocytogenes in chill brine to below detectable levels after enrichment. Fresh brine solution was inoculated with L. monocytogenes and exposed to UV and/or antimicrobial agent at −1C in a recirculating UV treatment unit. When L. monocytogenes was no longer detectable via direct plating on MOX, enrichment in brain–heart infusion broth was performed, and suspect colonies were confirmed using API Listeria. The combinations of UV + 0.5% CA and UV + 500-ppm DMDC were found to be the most effective, where L. monocytogenes was undetectable via enrichment at 45 and 60 min of treatment, respectively. CA (0.5%) when used in the absence of UV resulted in nondetection of L. monocytogenes. However, the reduction rate was higher when UV was used concurrent with CA. This work indicates that combinations of UV and antimicrobials may be more effective than either of the treatments alone for the reduction of L. monocytogenes in fresh brines. PRACTICAL APPLICATIONS This paper describes the use of UV light and antimicrobial agents to control L. monocytogenes in chilled brines used for product cooling in the ready-to-eat meat industry. The data include time of exposure to UV light and level of antimicrobial agent, which result in inactivation of the pathogen. These data may support industry efforts to validate the use of UV light and antimicrobial agents to control L. monocytogenes in recirculating chill brine systems.

Fresh and fresh-cut tomatoes have been associated with numerous outbreaks of salmonellosis in recent years. One effective post harvest treatment to reduce Salmonella enterica in tomatoes may be high pressure processing (HPP). The objectives of the study were to determine the potential for HPP to reduce S. enterica serovars Newport, Javiana, Braenderup and Anatum in tryptic soy broth (TSB) and to determine the effect of HPP to reduce the most pressure resistant of the four serovars from fresh diced and whole tomatoes. To evaluate pressure resistance, TSB containing 8 log CFU/ml of one of the four serovars was packaged in sterile stomacher bags and subjected to one of three different pressures (350, 450 or 550MPa) for 120s. The most pressure resistant S. enterica serovar evaluated was Braenderup. Subjecting the broth culture to 350, 450 and 550MPa resulted in a 4.53, 5.74 and 7.09 log reduction in S. Braenderup, respectively. Diced tomatoes (150g) and whole red round tomatoes (approximately 150g) were inoculated with 0.1ml of 9.1 log CFU/ml S. Braenderup, and subjected to the same pressure treatments (350, 450 or 550MPa). Significant reductions of S. Braenderup concentrations in diced tomatoes (P0.05) were seen after processing at 350 (0.46CFU/g), 450 (1.44 log CFU/g), and 550MPa (3.67 log CFU/g). In whole tomatoes, significant reductions (P0.05) were also seen at 350 (1.41 log CFU/g), 450 (2.25 log CFU/g) and 550MPa (3.35 log CFU/g). HPP may be an effective post harvest strategy to reduce low levels of S. enterica contamination in whole and diced tomatoes.

Environmental factors encountered during growing and harvesting may contribute to Escherichia coli O157:H7 contamination of lettuce. Limited nutrients and extended exposure to water may cause E. coli O157:H7 to shed its O antigen. Absence of the O157-polysaccharide antigen could affect the cell's physicochemical properties (hydrophobicity and cell charge) and ultimately influence its attachment to surfaces. The objectives of this study were to evaluate the effect of the E. coli O157:H7 O-antigen on the cell's overall hydrophobicity, charge and ability to attach to cut edge and whole leaf iceberg lettuce surfaces. Three strains of E. coli O157:H7 (86-24 wild type; F-12, mutant lacking the O-antigen and pRFBE, plasmid for O157 gene reintroduced) were examined for their hydrophobicity, overall charge and ability to attach to lettuce. Overall, E. coli O157:H7 attached at higher levels to cut surfaces over whole leaf surfaces (P=0.008) for all strains and treatments. Additionally, the strain lacking the O-antigen (F12)-attached significantly less to lettuce (P=0.015) than the strains expressing the antigen (WT and pRFBE). Cells lacking the O antigen (strain F-12) were also significantly more hydrophobic than strains 86-24 or pRFBE (P≤0.05). Surface charge differed among the strains tested (P≤0.05); however, it did not appear to influence bacterial attachment to lettuce surfaces. The charge was not fully restored in the pRFBE strain (expression of O-antigen reintroduced), therefore, no conclusions can be made pertaining to the effect of charge on attachment in this study. Results indicate that E. coli O157:H7 cells which lack the O-antigen have greater hydrophobicity and attach at lower concentrations than cells expressing the O-antigen, to iceberg lettuce surfaces.

The effects of packaging treatments, postharvest cooling delay and storage duration on color, texture, ascorbic acid content, weight loss and glucosinolate retention in crown-cut heads of broccoli were studied. Broccoli stored in shrink wrap film lost 3.7% of its original weight, whereas ice packaging resulted in 17.4% weight loss during storage. Long postharvest cooling delay and storage duration negatively affected broccoli color. Broccoli heads stored in shrink wrap packaging retained firmness longer than broccoli stored in ice. Ascorbic acid retention was improved in shrink wrapped broccoli, but retention decreased as postharvest cooling delay and storage duration lengthened. Shrink wrapped broccoli exhibited improved retention of the glucosinolate, glucoraphanin. During storage, “Gypsy” broccoli maintained better quality than “Everest” with respect to color, ascorbic acid retention and weight loss. However, “Everest” retained texture (firmness) better after 35 days of storage. Shrink wrap packaging and shorter postharvest cooling delays preserve broccoli quality and increase shelf life. PRACTICAL APPLICATIONS Broccoli has the potential to be a high value crop if the shrink wrap packaging technology can be disseminated to the broccoli growers. Typically, farmers use ice for cooling broccoli heads after harvest, which is an expensive and difficult practice. Without icing requirement, broccoli marketing will be easy and opens up significant potential for broccoli producers who can afford relatively inexpensive film wrapping machines (as compared with ice equipment) in conjunction with onsite cooling. Sometimes onsite cooling is not possible and delays may be involved before the harvested produce is cooled. The conclusions reached in this study shows that shrink wrap packaging can be used to improve quality, retain phytochemical content and prolong the shelf life of crown-cut broccoli.

This study investigated the effects of packaging and storage temperature on the spinach phylloepiphytic bacterial community and fate of Escherichia coli O157:H7. Freshly harvested spinach was rinsed and/or disinfected, packaged and stored under typical retail conditions (4 degrees C) or under temperature abuse conditions (10 degrees C) for a period of 15 days. The final population size of culturable epiphytic bacteria after 15 days of storage was not affected by the temperature of storage or the presence of E. coli O157:H7. However, analysis of the bacterial community using denaturing gradient gel electrophoresis of 16s rDNA revealed changes with time of storage and the presence of E. coli O157:H7. Excision and sequencing of prominent DGGE bands identified that the majority of sequences belonged to the phyla Actinobacteria, Bacteroidetes, Firmicutes and Alphaprotebacteria. After 10 days of storage at 4 degrees C or 10 degrees C the population became more dominated by psychrotrophic bacteria. Removal of the epiphytic bacteria resulted in significant increases in numbers of E coli O157:H7 at 10 degrees C and was associated with decreased expression of E. coli O157:H7 virulence (stxA, curli, eaeA) and stress response (rpoS, sodB) genes. In conclusion, storage temperature and time of storage of packaged spinach affected the diversity of the epiphytic spinach microbiota which influenced the growth, establishment, physiology and potentially virulence of E. coli O157:H7.

Cinnamic acid (CA), a naturally occurring organic acid found in fruits and spices, has antimicrobial activity against spoilage and pathogenic bacteria, but low aqueous solubility limits its use. The purpose of this study was to determine the effectiveness of solubility-enhancing alpha-cyclodextrin-CA inclusion complexes against Escherichia coli O157:H7 and Salmonella enterica serovars suspended in apple cider or orange juice at two different incubation temperatures (4 and 26 degrees Celsius). Two concentrations (400 and 1,000 mg/liter) of alpha-cyclodextrin-CA inclusion complex were aseptically added to apple cider inoculated with E. coli O157:H7 (7 log CFU/ml) and orange juice inoculated with a cocktail of six Salmonella enterica serovars (7 log CFU/ml). Samples were extracted at 0 min, at 2 min, and at 24-h intervals for 7 days, serially diluted in 0.1 % peptone, spread plated in duplicate onto tryptic soy agar, and incubated at 35 degrees Celsius for 24 h. Populations of E. coli O157:H7 in apple cider were significantly reduced (Por = 0.05) during the 7-day sampling period in all solutions regardless of temperature. Compared with the controls, populations were significantly reduced by the addition of 400 and 1,000 mg/liter inclusion complex, but reductions were not significantly different (Por = 0.05) between the two treatment groups (400 and 1,000 mg/liter). Salmonella was significantly reduced in all solutions regardless of temperature. There were significant differences between the control and each inclusion complex concentration at 4 and 26 degrees Celsius. Coupled with additional processing steps, alpha-cyclodextrin-CA inclusion complexes may provide an alternative to traditional heat processes.

The purpose of this research was to establish the dose of UV light (253.7 nm) needed to inactivate Listeria monocytogenes in distilled water, fresh brine (9% NaCl), spent brine, and diluted (5, 35, and 55%) spent brine, using uridine as a chemical actinometer. Strains N1-227 (isolated from hot dog batter), N3-031 (isolated from turkey franks), and R2-499 (isolated from meat) were mixed in equal proportions and suspended in each solution prepared so as to contain 10 ―4 M uridine. Samples were irradiated in sterile quartz cells for 0, 5, 10, 15, 20, 25, or 30 min. Inactivation was evaluated by serially diluting samples in 0.1 % peptone, by surface plating in duplicate onto modified Oxford agar and Trypticase soy agar with yeast extract, and by enrichment in brain heart infusion broth, followed by incubation at 37°C for 24 to 48 h. For dose measurements, the absorbance (262 nm) was measured before and after imadiation. Differences were observed in population estimates depending on the solution (P ≤ 0.05). Reductions were as follows from greatest to least: water > fresh brine > 5% spent brine > 35% spent brine > 55% spent brine > undiluted spent brine. UV light did not significantly reduce populations suspended in spent brine solutions. L. monocytogenes decreased to below the detection limit (1 log CFU/ml) at doses greater than 33.2 mJ/cm 2 in water and at doses greater than 10.3 mJ/cm 2 in fresh brine. Knowledge of UV dosing required to control L. monocytogenes in brines similar to those used for ready-to-eat meat processing will aid manufacturers in establishing appropriate food safety interventions for these products.

Tomatoes have been linked to outbreaks of salmonellosis, demonstrating the need to identify sources of contamination. Objectives of this study included determining the ability for Salmonella enterica serovar Montevideo to be internalized into tomatoes from contaminated irrigation water and seed stock, and establishing whether Salmonella Montevideo can survive in fertilizer solutions. Six treatment groups (five plants per group) were irrigated with 350 ml of 7 log CFU/ml of Salmonella Montevideo every 14 days for 70 days, each group receiving an increased number of contaminated water events progressively: group 1 received one contaminated watering at day 0, and group 6 received a total of six contaminated waterings. Group 7 was a control, and group 8 was grown from seeds soaked in 8 log CFU/ml of Salmonella Montevideo for 24 h. All plants were watered daily with uncontaminated water. Three replications were completed. Fruit from every plant, and roots, stems, and leaves of one plant per treatment were sampled. All tomatoes were negative for Salmonella Montevideo; five root samples tested positive. For fertilizer studies, a commercially available fertilizer, two custom mixed and 1.0% dilutions of each (total of six solutions), and sterile water were inoculated with 8 log CFU/ml of Salmonella Montevideo and stored at 25 degrees C. Solutions were sampled at 24, 48, and 72 h. There were no differences (Por = 0.05) between survival of Salmonella Montevideo in diluted fertilizers and the control. Results indicate Salmonella Montevideo is unable to contaminate tomato fruit via irrigation water and seed stock but can survive in fertilizer solutions.

Ready-to-eat meat products have been implicated in several foodborne listeriosis outbreaks. Microbial contamination of these products can occur after thermal processing when products are chilled in salt brines. The objective of this study was to evaluate UV radiation on the inactivation of Listeria monocytogenes and lactic acid bacteria in a model brine chiller system. Two concentrations of brine (7.9% [wt/wt] or 13.2% [wt/wt]) were inoculated with a approximately 6.0 log CFU/ml cocktail of L. monocytogenes or lactic acid bacteria and passed through a UV treatment system for 60 min. Three replications of each bacteria-and-brine combination were performed and resulted in at least a 4.5-log reduction in microbial numbers in all treated brines after exposure to UV light. Bacterial populations were significantly reduced after 5 min of exposure to UV light in the model brine chiller compared with the control, which received no UV light exposure (P0.05). The maximum rate of inactivation for both microorganisms in treated brines occurred between minutes 1 and 15 of UV exposure. Results indicate that in-line treatment of chill brines with UV light reduces the number of L. monocytogenes and lactic acid bacteria.

The effect of high pressure on the log reduction of six strains of Escherichia coli O157:H7 and five serovars of Salmonella enterica was investigated in tryptic soy broth, sterile distilled water, and commercially sterile orange juice (for Salmonella) and apple cider (for E. coli). Samples were subjected to high-pressure processing treatment at 300 and 550 MPa for 2 min at 6 degrees C. Samples were plated onto tryptic soy agar directly after pressurization and after being held for 24 h at 4 degrees C. At 300 MPa, little effect was seen on E. coli O157:H7 strains, while Salmonella serovars varied in resistance, showing reductions between 0.26 and 3.95 log CFU/ml. At 550 MPa, E. coli O157:H7 strains exhibited a range of reductions (0.28 to 4.39 log CFU/ml), while most Salmonella populations decreased beyond the detection limit (5-log CFU/ml reduction). The most resistant strains tested were E. coli E009 and Salmonella Agona. Generally, bacterial populations in fruit juices showed larger decreases than did populations in tryptic soy broth and distilled water. E. coli O157:H7 cultures held for 24 h at 4 degrees C after treatment at 550 MPa showed a significant log decrease as compared with cultures directly after treatment (Por = 0.05), while Salmonella serovars did not show this significant decrease (P0.05). All Salmonella serovars tested in orange juice treated at 550 MPa for 2 min at 6 degrees C and held for 24 h showed a5-log decrease, while E. coli O157:H7 strains require a higher pressure, higher temperature, longer pressurization, or a chemical additive to achieve a 5-log decrease.

Curli fibers are produced by some Escherichia coli cells in response to environmental stimuli. These extracellular proteins enhance the cell's ability to form biofilms on various abiotic surfaces. E. coli 0157:H7 cells readily attach to a variety of fruit and vegetable surfaces. It is not known whether the expression of curli influences the cell's ability to attach to produce surfaces. In this study, the effect of curli expression on the cell's overall hydrophobicity, charge, and ability to attach to cut and whole iceberg lettuce surfaces was examined. All strains, regardless of curli expression, attached preferentially to the cut edges of lettuce (P < 0.05). The curli-producing cells of E. coli 0157:H7 strain E0018 attached in significantly greater numbers to both cut and whole lettuce pieces than did the non-curli-producing E0018 cells (P < 0.05); however, no significant attachment differences were observed between the curli-producing and non-curli-producing cells of E. coli 0157:H7 strains 43894 and 43895. All curli-producing E. coli 0157:H7 strains were significantly more hydrophobic (P < 0.01); however, no association between the cells' hydrophobic characteristics and lettuce attachment was observed. Overall surface charge of the cells did not differ among strains or curli phenotypes. Results indicate that overall hydrophobicity and cell charge in E. coli 0157:H7 strains do not influence attachment to iceberg lettuce surfaces. The presence of curli may not have any influence on attachment of E. coli 0157:H7 cells to produce items. Additional factorsmay influence the attachment of E. coli 0157:H7 to plant surfaces and should be further examined.

The ability of Listeria monocytogenes to attach to various food contact surfaces, such as stainless steel, polypropylene, and rubber compounds, is well documented. The retention of these or other pathogenic bacteria on food contact surfaces increases the risk of transmission to food products. The objective of this study was to compare several methods for quantitative recovery of Listeria monocytogenes from stainless steel surfaces. A cocktail of 4 serotypes of Listeria monocytogenes mixed in equivalent concentrations was inoculated onto type 304 stainless steel coupons in a 2 2 cm area. After 1 h exposure, coupons were sampled by one of the following methods: (1) swabbing with a premoistened Dacron swab; (2) rinsingwith phosphate-buffered saline; (3) direct contact onto tryptic soy agar containing 0.6% yeast extract (TSA + YE) plates for 10 s; (4) sonication in an ultrasonic water bath (40 kHz); (5) contact with the bristles of a sonicating brush head for 1 min; and (6) indirect contact (24 mm distance) with a sonicating brush head for 1 min. The 3 sonication methods yielded higher recovery than the other 3 methods (P < 0.05). Brushing the coupons with the sonicating brush head (contact or noncontact) yielded a recovery level of about 60%. The lowest cell recovery (about 20%) was observed with the swab and direct agar contact methods. After a 12 h exposure, recoveries ranged from 17.4 (brush contact method) to 2% (swab method).

Environmental sampling has become increasingly important in the food industry for monitoring the presence of specific pathogens such as Listeria monocytogenes and Salmonella enterica. Several microbiological media are available for storage and transport of environmental samples from the processing plant to the test laboratory. In this study, we quantified the survival of L. monocytogenes, S. Typhimurium, S. Enteritidis, and S. Typhi in environmental sampling media over several time and temperature combinations to determine optimum conditions for transport and storage. A cocktail of L. monocytogenes strains and Salmonella serotypes was separately added to tubes of Dey-Engley (D/E) Neutralizing Broth, Copan SRK solution, and Neutralizing Buffer and incubated at either –4, 4, 10, or 15C. Counts were made of the bacterial load after 0, 12, 24, and 48 h. Neutralizing Buffer and Copan SRK solution were best at maintaining bacterial concentrations at all temperatures. D/E Neutralizing Broth, at 10 and 15C, allowed significant bacterial growth. This study helped validate the use of these 3 media for environmental sample transport and storage at cold holding temperatures and demonstrated that, at elevated temperatures (>4C), it is preferable to use Neutralizing Buffer or Copan SRK solution for quantifying microbial recovery. E nvironmental sampling is a very important tool for studying the source, level, and dispersion of microbiological contamination in a defined area or facility. Two major microbial pathogens of concern in the food industry are Listeria monocytogenes and Salmonella spp. These 2 pathogens can be found in food processing facilities and are major concerns for public health officials. In the food industry, environmental sampling has become a common method for detecting microbial pathogens such as

Fresh blueberries are commonly stored and transported by refrigeration in controlled atmospheres to protect shelf life for long periods of storage. Ozone is an antimicrobial gas that can extend shelf life and protect fruit from microbial contamination. Shelf life of fresh highbush blueberries was determined over 10-day storage in isolated cabinets at 4°C or 12°C under different atmosphere conditions, including air (control); 5% O2 : 15% CO2 : 80% N2(controlled atmosphere storage (CAS)); and ozone gas (O3) 4 ppm at 4°C or 2.5 ppm at 12°C, at high relative humidity (90–95%). Samples were evaluated for yeast and molds growth, weight loss, and firmness. CAS and O3did not delay or inhibit yeast and molds growth in blueberries after 10 days at both temperatures. Fruit stored at 4°C showed lower weight loss values compared with 12°C. Blueberries stored under O3atmosphere showed reduced weight loss at 12°C by day 10 and loss of firmness when compared to the other treatments. Low concentrations of ozone gas together with proper refrigeration temperature can help protect fresh blueberries quality during storage.

Flavor characteristics of roasted peanuts over short-term storage were explored through sensory and chemical analyses. The volatiles from freshly roasted peanuts were evaluated over short-time (21 d) storage using gas chromatography, chemosensory techniques, and a sensory panel to quantify and identify pyrazines and hexanal over a 21-d storage period. A significant decrease (P 0.05) was noted in the 2-methylpyrazine and 2-ethylpyrazine concentrations. The hexanal concentration significantly increased (P < 0.05) over the 21-d period. The peroxide values and sensory analysis agreed with these results. A significant increase (P < 0.05) in peroxide value was seen at days 14 and 21, and a significant decrease (P < 0.05) in fresh roasted peanuty flavor from day 0 to 21 and significant increases (P < 0.05) in painty, cardboardy, and bitter flavors from day 7 to 21 with the sensory analysis. The electronic nose successfully separated day 0 and 21 samples from day 7 and 14 samples, which were also separated but with some overlap.

Inactivation of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice treated with ozone in combination with antimicrobials was evaluated. E. coli O157:H7 or Salmonella was suspended in cider and orange juice, and ozone was pumped into juices (4°C) containing dimethyl dicarbonate (DMDC; 250 or 500 ppm) or hydrogen peroxide (300 or 600 ppm) for up to 90 min (study 1) or 60 min followed by 24-h storage at 4°C (study 2). Study 1: No combination of treatments resulted in a 5-log colony-forming units (CFU) /mL reduction of either pathogen. Study 2: All combinations of antimicrobials plus ozone treatments, followed by refrigerated storage, caused greater than a 5-log CFU/mL reduction, except ozone/DMDC (250 ppm) treatment in orange juice. Ozone treatment in combination with DMDC or hydrogen peroxide followed by refrigerated storage may provide an alternative to thermal pasteurization to meet the 5-log reduction standard in cider and orange juice.

Inactivation of Escherichia coli O157:H7 suspended in salt solutions (Na(2)PO(4), NaCl, NaNO(3), NH(4)Cl, CaCl(2), and AlCl(3)) at concentrations ranging from 0 to 5% (w/v) by high-intensity ultrasound at two different temperature conditions (ice water bath, 40 degrees C water bath) and three ultrasonic intensity levels (9.5, 21.8, 49.2 W/cm(2)) was determined. Increases in sonication treatment time, intensity, and temperature led to increased lethality of E. coli O157:H7. However, cell lethality, as a function of solute concentration, varied depending upon type of salt. CaCl(2), NaCl, NaNO(3), Na(2)PO(4), and NH(4)Cl had little or no effect on survival of E. coli O157:H7 (1-log reduction) regardless of other treatment conditions at 9.5 W/cm(2). Concentrations of0.5% of CaCl(2), NaNO(3), Na(2)PO(4), and NH(4)Cl adversely affected survival (1.0- to 1.6-log reduction) with treatment of 21.8 W/cm(2) in an ice bath and showed greater inactivation (1.2-4.0-log reduction) in the 40 degrees C water bath. Treatment of 49.2 W/cm(2) showed the greatest impact regardless of other parameters. In the 40 degrees C water bath, treatment for 10 min at 49.22 W/cm(2) led to total inactivation for cells suspended in 0.5% NH(4)Cl, 1% CaCl(2), 2% NaCl, 5% Na(2)PO(4), and all concentrations of AlCl(3). Complete inactivation also occurred in an ice bath for 5% AlCl(3), which was shown to be the most effective salt. The most efficient treatment combination was in the 40 degrees C water bath for 10 min at 49.2 W/cm(2) with 5% AlCl(3). Inactivation of E. coli O157:H7 is attributed to cavitation-induced shear forces, reaction of cavitation-generated hydrogen peroxide with microbial cell wall constituents and electrostatic interactions of dissociated salts with cell membranes.

The low water activity (a(w) 0.3) of peanut butter prohibits the growth of Salmonella in a product; however, illnesses are reported from peanut butter contaminated with very small doses, suggesting the food matrix itself influences the infectious dose of Salmonella, potentially by improving Salmonella's survival in the gastrointestinal tract. The purpose of our study was to quantify the survival of a peanut butter outbreak-associated strain of Salmonella enterica serotype Tennessee when inoculated into peanut butters with different fat contents and a(w) (high fat, high a(w); high fat, low a(w); low fat, high a(w); low fat, low a(w)) and then challenged with a simulated gastrointestinal system. Exposures to increased fat content and decreased a(w) both were associated with a protective effect on the survival of Salmonella Tennessee in the simulated gastric fluid compared with control cells. After a simulated intestinal phase, the populations of Salmonella Tennessee in the control and low-fat formulations were not significantly different; however, a 2-log CFU/g increase occurred in high-fat formulations. This study demonstrates that cross-protection from low-a(w) stress and the presence of high fat results in improved survival in the low pH of the stomach. The potential for interaction of food matrix and stress adaptations could influence the virulence of Salmonella and should be considered for risk analysis.

Exposure to sublethal processing treatments can stimulate bacterial stress responses. The purpose of this research was to determine whether adaptation to common food processing stresses encountered during the preparation of ready-to-eat foods affects the dose of UV light required to significantly reduce Listeria monocytogenes populations in sterile distilled water and a 9% NaCl solution, using uridine as a chemical actinometer. L. monocytogenes strains N1-227 (from hot dog batter), N3-031 (from turkey franks), and R2-499 (from ready-to-eat meat) were acid stressed for 3 h at 35 degrees C in Trypticase soy broth with yeast extract acidified to pH 5.0, heat shocked for 1 h at 48 degrees C in brain heart infusion broth (BHIB), and selected for sulfanilamide resistance (512 microg/ml). These strains were then mixed in equal proportions and suspended in water and 9% NaCl solution, each containing 10(-4) M uridine. Samples were exposed to UV light (253.7 nm) for 0, 5, 10, 15, 20, 25, or 30 min. Inactivation was evaluated by surface plating onto modified Oxford agar and Trypticase soy agar with yeast extract and by enrichment in BHIB followed by incubation at 37 degrees C for 24 h. The absorbance of each sample was measured before and after irradiation to calculate the dose of UV light. There were no significant differences between population estimates based on medium or suspension solution. There were no population differences between acid-stressed and antibiotic-resistant or unstressed and heat-shocked L. monocytogenes strains. However, acid-stressed and antibiotic-resistant strains were significantly more resistant to UV light than were unstressed and heat-shocked strains (P < or = 0.05).

Listeria monocytogenes is the pathogen of concern in ready-to-eat (RTE) meat products. Salt brines are used to chill processed meats. L. monocytogenes and lactic acid bacteria (LAB) can grow under saline conditions, and may compete with each other for nutrients. The objective of this study was to determine the effect of lactic acid bacteria (Enterococcus faecalis, Carnobacterium gallinarum, and Lactobacillus plantarum) on the survival of L. monocytogenes and Listeria innocua in brines stored under low temperatures for 10 d. Sterile tap water (STW) and 2 brine solutions (7.9% and 13.2% NaCl) were inoculated with 1 of 5 cocktails (L. monocytogenes, L. innocua, LAB, L. monocytogenes+ LAB, or L. innocua+ LAB) at initial concentrations of 7 log CFU/mL. Brines were stored for 10 d at 4 or 12 degrees C. Three replications of each brine concentration/cocktail/temperature combination were completed. No significant reductions of L. monocytogenes occurred in 7.9%[w/v] or 13.2%[w/v] brines when LAB were present; however, there were significant reductions after 10 d of L. monocytogenes in the STW solution when LAB were present (1.43 log CFU/mL at 4 degrees C and 3.02 log CFU/mL at 12 degrees C). L. innocua was significantly less resilient to environmental stresses of the brines than L. monocytogenes, both with and without LAB present (P< or = 0.05). These strains of lactic acid bacteria are not effective at reducing L. monocytogenes in brines at low temperatures. Furthermore, use of L. innocua as a model for L. monocytogenes is not appropriate under these environmental conditions.

The effect of high pressure processing in conjunction with the chemical antimicrobials, dimethyl dicarbonate (DMDC), hydrogen peroxide, cinnamic acid, potassium sorbate, and sodium benzoate (NaB) on E. coli O157:H7 strain E009 and Salmonella enterica serovar Agona was investigated in apple juice and orange juice, respectively. Juices were inoculated with approximately 10(6) CFU/ml and subjected to pressures of 550 MPa (E. coli O157:H7 samples) and 400 MPa (Salmonella Agona samples) for 2 min at 6 degrees C (initial temperature). Populations of each pathogen were determined before pressurization, immediately after pressurization, and after samples had been held after treatment for 24 h at 4 degrees C. The most effective treatment for E. coli O157:H7, as determined by plating immediately after pressurization, was 125 ppm of DMDC, which caused a >4.98-log reduction. Other treatments that were significantly different from the sample with no added antimicrobial were 62.5 ppm of DMDC, 300 ppm of hydrogen peroxide, and 500 ppm of NaB, which produced 4.97-, 5.79-, and 3.91-log total reductions, respectively. After 24 h at 4 degrees C, E. coli O157:H7 was undetectable in all treatment groups (and controls). In samples inoculated with Salmonella, the most effective treatment was 62.5 ppm of DMDC, which produced a 5.96-log decrease immediately after pressure treatment. The results for 1,000 ppm of NaB, which produced a 3.26-log decrease, also were significantly different from those for the sample containing no antimicrobials. After 24 h at 4 degrees C, all samples with added antimicrobials had near or more than a 5-log total reduction of Salmonella Agona.

Packaging fishery products under vacuum atmosphere packaging (VAC) and modified atmosphere packaging (MAP) conditions can significantly extend the shelf life of raw, refrigerated fish products. There is considerable commercial interest in marketing VAC and MAP refrigerated (never frozen) raw fish fillets. The objective of this study was to determine if Clostridium botulinum toxin development precedes microbiological spoilage in raw, refrigerated flounder fillets. Aquacultured flounder (Paralichthys dentatus) individual fish fillets either were packed with a film having an oxygen transmission rate (OTR) of 3,000 cm3 m-2 24 h-1 at 22.8°C or were vacuum packaged or packaged under 100% CO2 with a film having an OTR of 7.8 cm3 m-2 24 h-1 at 21.1°C and were stored at 4 and 10°C. Samples were analyzed by aerobic plate count (APC) for spoilage and qualitatively for botulinum toxin with a mouse bioassay. The results demonstrate that flounder fillets (4°C) packaged with a film having an OTR of 3,000 were microbiologically spoiled (APC, >107 CFU/g) on day 15, but there was no toxin formation, even after 35 days of storage. However, at 10°C, toxin production occurred (day 8), but it was after microbial spoilage and absolute sensory rejection (day 5). Vacuum-packaged fillets and 100% CO2 fillets (4°C) packaged with a film having an OTR of 7.8 were toxic on days 20 and 25, respectively, with microbial spoilage (APC, >107 CFU/g) not occurring during the tested storage period (i.e., >35 days). At 10°C, in vacuum-packaged flounder, toxin formation coincided with microbiological spoilage (days 8 to 9). In the 100% CO2-packaged fillets, toxin formation occurred on day 9, with microbial spoilage occurring on day 15. This study indicates that films with an OTR of 3,000 can be used for refrigerated fish fillets and still maintain the safety of the product. Published version

The fate of Yersinia enterocolitica was investigated during the manufacture and storage of yogurt produced using commercial starter cultures YC470 (rapid acid-producer), YC180 (slow acid-producer), both containing Streptococcus thermophilus and Lactobacillus bulgaricus, and a “home-use” culture that contained these bacteria as well as Lactobacillus acidophilus. Preparations were fermented at 44°C and stored at 4°C. During the first few hours of fermentation, Y. enterocolitica populations increased in all preparations. Y. enterocolitica populations were reduced to undetectable levels after 72 and 96h in YC470 and YC180 yogurt, respectively, but survived up to 144h in “home-use” yogurt. The home-use starter produced lactic acid in greater amounts and more rapidly (p

Inactivation of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice treated with ozone was evaluated. A five-strain mixture of E. coli O157:H7 or a five-serovar mixture of Salmonella was inoculated (7 log CFU/ml) into apple cider and orange juice. Ozone (0.9 g/h) was pumped into juices maintained at 4 degrees C, ambient temperature (approximately 20 degrees C), and 50 degrees C for up to 240 min, depending on organism, juice, and treatment temperature. Samples were withdrawn, diluted in 0.1% peptone water, and surface plated onto recovery media. Recovery of E. coli O157:H7 was compared on tryptic soy agar (TSA), sorbitol MacConkey agar, hemorrhagic coli agar, and modified eosin methylene blue agar; recovery of Salmonella was compared on TSA, bismuth sulfite agar, and xylose lysine tergitol 4 (XLT4) agar. After treatment at 50 degrees C, E. coli O157:H7 populations were undetectable (limit of 1.0 log CFU/ml; a minimum 6.0-log CFU/ml reduction) after 45 min in apple cider and 75 min in orange juice. At 50 degrees C, Salmonella was reduced by 4.8 log CFU/ml (apple cider) and was undetectable in orange juice after 15 min. E. coli O157:H7 at 4 degrees C was reduced by 4.8 log CFU/ml in apple cider and by 5.4 log CFU/ml in orange juice. Salmonella was reduced by 4.5 log CFU/ml (apple cider) and 4.2 log CFU/ml (orange juice) at 4 degrees C. Treatment at ambient temperature resulted in population reductions of less than 5.0 log CFU/ml. Recovery of E. coli O157:H7 and Salmonella on selective media was substantially lower than recovery on TSA, indicating development of sublethal injury. Ozone treatment of apple cider and orange juice at 4 degrees C or in combination with mild heating (50 degrees C) may provide an alternative to thermal pasteurization for reduction of E. coli O157:H7 and Salmonella in apple cider and orange juice.

The effect of package atmosphere on survival of uninjured and sublethally heat-injured Listeria monocytogenes, inoculated onto tryptose phosphate agar containing 0.85% lactic acid and 2% NaCl (TPALAS) was investigated. Inoculated TPALAS plates were packaged in air, 100% N2 (N2), 30% CO2-70% N2 (CO2-N2), and vacuum and stored at 4 and 20 degrees C for up to 31 days. Recovery of L. monocytogenes from TPALAS was influenced by the injury status (i.e., injured and uninjured) of the inoculum, storage atmosphere (air, N2, CO2-N2, and vacuum), storage temperature (4 and 20 degrees C), and recovery media [tryptose phosphate agar (TPA) and modified Oxford agar (MOX)] (P0.05). Overall, storage at 4 degrees C supported greater survival than storage at 20 degrees C (P0.05). Uninjured L. monocytogenes stored at 4 degrees C was recovered on TPA better than sublethally heat-injured L. monocytogenes stored at 40 degrees C (P0.05). Recovery of sublethally heat-injured L. monocytogenes stored at 4 degrees C followed the order N2CO2-N2airvacuum (P0.05), whereas recovery of uninjured L. monocyrogenes stored at 4 degrees C followed the order N2CO2-N2vacuumair (P0.05). Air and vacuum atmospheres supported greater survival of uninjured and heat-injured L. monocytogenes than N2 and CO2-N2 atmospheres at 20 degrees C (P0.05). Recovery of sublethally heat-injured L. monocytogenes stored at 20 degrees C followed the order vacuumairCO2-N2 = N2 (P0.05), whereas recovery of uninjured L. monocytogenes stored at 20 degrees C followed the order vacuumairCO2-N2N2 (P0.05). Uninjured L. monocytogenes stored under N2 at 4 degrees C was recovered best, whereas sublethally heat-injured L. monocytogenes stored under N2 at 20 degrees C was recovered poorest (P0.05). Factors such as package atmosphere and storage temperature, involved in the production, storage, and distribution of fermented foods must be thoroughly evaluated when determining strategies for control and detection of L. monocytogenes in such products.

The effect of atmospheric composition and storage temperature on growth and survival of uninjured and sublethally heat-injured Escherichia coli O157:H7, inoculated onto brain heart infusion agar containing 0.3% beef extract (BEM), was determined. BEM plates were packaged in barrier bags in air, 100% CO2, 100% N2, 20% CO2 : 80% N2, and vacuum and were stored at 4, 10, and 37°C for up to 20 days. Package atmosphere and inoculum status (i.e., uninjured or heat-injured) influenced (P < 0.01) growth and survival of E. coli O157:H7 stored at all test temperatures. Growth of heat-injured E. coli O157:H7 was slower (P < 0.01) than uninjured E. coli O157:H7 stored at 37°C. At 37°C, uninjured E. coli O157:H7 reached stationary phase growth earlier than heat-injured populations. Uninjured E. coli O157:H7 grew during 10 days of storage at 10°C, while heat-injured populations declined during 20 days of storage at 10°C. Uninjured E. coli O157:H7 stored at 10°C reached stationary phase growth within approximately 10 days in all packaging atmospheres except CO2. Populations of uninjured and heat-injured E. coli O157:H7 declined throughout storage for 20 days at 4°C. Survival of uninjured populations stored at 4°C, as well as heat-injured populations stored at 4 and 10°C, was enhanced in CO2 atmosphere. Survival of heat-injured E. coli O157:H7 at 4 and 10°C was not different (P > 0.05). Uninjured and heat-injured E. coli O157:H7 are able to survive at low temperatures in the modified atmospheres used in this study.Key words: E. coli O157:H7, sublethal injury, modified atmosphere packaging.