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During the early stages of aerobic composting, heat is generated and when the materials are self-insulating, extended exposure of pathogens to this heat source will lead to significant reduction, if not elimination, of the pathogens. However, when insufficient heat is applied to the composting materials, pathogens may survive. Under those conditions if the compost had contained material of animal origin or food waste, it would be considered untreated and would not be allowed in fields growing crops that may be consumed raw. However, alternative treatment processes are allowed, provided they are validated to meet the microbial standards stipulated in the Produce Safety final rule of the Food Safety Modernization Act and that the physical parameters of the process are documented to ensure that the conditions under which the process was validated have been met. Hence, this exploratory study was undertaken in a laboratory setting to determine the potential for application of aerosolized smoke to inactivate Salmonella in manure-based compost. Smoke generated from wood chips (oak or pecan) and introduced to the headspace of contaminated cow manure compost (≤3 log CFU/g) in sealed containers (35 g per container) resulted in no Salmonella detected by enrichment culture in 100% (0 of 14) of the samples after 18 to 48 h of exposure, whereas Salmonella in control samples remained at initial levels over the same time period. Shorter exposure times (6 h) to the smoke aerosols were less effective (11 of 24 samples positive by enrichment culture), and additional flushes with the wood smoke during this time failed to decrease the prevalence of contamination. Smoke aerosols generated from waste agricultural materials and held in containers with Salmonella -contaminated compost for 18 h significantly reduced the prevalence of the pathogen in samples compared with control samples ( P < 0.05). The odds of not finding Salmonella in smoke-exposed compost were 14 (pine needles and rice hulls), 23 (cocoa hulls, orange rind, and peanut hulls), and 28 (sunflower hulls) times greater compared with samples not exposed to smoke. Many other variables remain to be examined (e.g., compost composition, compost maturity, and anaerobic conditions) to determine whether this approach could be universally applied to manure-based compost. Validation under field conditions will be required and may entail use of this approach in combination with suboptimal thermal conditions (<55°C).

Outbreaks associated with fresh-cut leafy greens continue to occur despite efforts to implement horticultural practices that minimize introduction of enteric pathogens to the crop. The experimental trials in this study were designed to examine the efficacy of an acetic acid (AA)- and chitosan-based spray treatment, applied 1 day prior to harvest, for reducing the prevalence of Escherichia coli O157:H7 (O157) and Salmonella in field-grown leafy greens contaminated at levels detectable only through enrichment culture. Responses to the treatment solution were variable and depended on the type of leafy green (leafy lettuce, spinach, or cabbage), cultivar, pathogen, and AA concentration (0.3 to 0.7%). No significant differences in E. coli O157 prevalence were found for untreated and treated cabbage heads and spinach plants ( P > 0.05). In contrast, treatment significantly affected Salmonella on 'Bravo F1' green cabbage and '7-Green' spinach ( P < 0.05), with odds ratios of 2.2 and 3.3 for finding the pathogen on untreated versus treated greens, respectively. Salmonella was also 7.1 times more likely to be found on an untreated lettuce plant than on a lettuce plant sprayed with a 0.7% AA treatment solution (95% confidence interval [CI], 4.1 to 12.2; P < 0.0001). In studies addressing the efficacy of chitosan (0.1 or 0.3%), this chemical failed to reduce the prevalence of either pathogen on lettuce ( P > 0.05). Similarly, spraying with 0.3% AA did not affect the prevalence of Salmonella on lettuce plants ( P > 0.05); however, treatment solutions with 0.4% AA reduced the likelihood of detecting Salmonella in treated versus untreated plants by 6.6 times (95% CI, 2.1 to 20.9; P = 0.0007). After the lettuce was harvested and hand washed, consumers failed to distinguish either visually or organoleptically between untreated lettuce and lettuce sprayed with an acetic acid solution ( P > 0.05). These results indicate that acetic acid could be used to reduce the microbiological risk of preharvest leafy greens.

Background: Cabbage may become contaminated with enteric pathogens during cultivation. Using multiple cabbage cultivars at two maturity stages (small plants or plants with small heads) in growth chamber studies, the fate (internalization or surface survival) of Salmonella and Escherichia coli O157:H7 (0157) were examined in conjunction with any potential relationships to the plant's antimicrobial content., Results: Internalized Salmonella was detected in cabbage within 24 h with prevalence ranging from 62% (16 of 26) for the 'Super Red 80' cultivar to 92% (24 of 26) for the 'Red Dynasty' cultivar. Surface survival of pathogens on small cabbage plants over nine days was significantly affected by cultivar with both pathogens surviving the most on the 'Farao' cultivar and Salmonella and O157 surviving the least on the 'Super Red 80' and 'Capture' cultivars, respectively (P < 0.05). Survival of O157 was slightly higher on cabbage heads for O157 than small plants suggesting that the maturity stage may affect this pathogen's fate. An inverse relationship existed between antimicrobial levels and the pathogen's surface survival on cabbage heads (P < 0.05)., Conclusions: The fate of pathogens varied with the cabbage cultivar in growth chamber studies highlighting the potential to explore cultivar in field studies to reduce the risk of microbiological contamination in this crop. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

To reduce the number of cabbage pathogen outbreaks, it is essential to understand the fate of enteric pathogens that contaminate plants in the field. To assist in that effort, two independent trials were conducted with a red cultivar (cv. Red Dynasty) and a green cultivar (cv. Bravo F1) of field-grown cabbage ( Brassica oleracea var. capitata). In the first trial, plants with small heads were sprayed with an inoculum containing both attenuated Salmonella enterica Typhimurium and Escherichia coli O157:H7 (5.0 log CFU/mL). Initial pathogen levels (ca. 3.9 log CFU per head), determined through plate count enumeration (limit of detection was 1.3 log CFU/g), dropped precipitously such that 2 days later, they could not be detected by enrichment culture in 22 to 35% of the heads. However, subsequent declines were at a slower rate; no differences were observed between red and green cabbage heads ( P > 0.05), and heads were still positive for the pathogens 22 days after being sprayed with the inoculum. As a result, the logistic model revealed that for every 2 days contaminated cabbage heads remained in the field, the probability of finding a positive sample decreased by a factor of 1.1 (95% confidence interval from 1.0 to 1.2, P = 0.0022) and 1.2 (95% confidence interval from 1.0 to 1.4, P ≤ 0.0001) for Salmonella and E. coli O157:H7, respectively. In the second trial occurring 2 weeks later, plants with medium red or green cabbage heads were sprayed with an inoculum at a dose of 3.5 log CFU/mL. A similar decay in prevalence over time occurred for green cabbage as in trial 1; however, pathogen decline in red cabbage was less in trial 2 than in trial 1. The extended persistence of pathogens in cabbage heads exhibited in both trials infers that harvest of contaminated cabbage destined for raw consumption is risky. Additional field studies are necessary to determine whether similar pathogen fates occur in other regions or climates and to clarify the effect of the maturity of red cabbage on pathogen inactivation.

Plant genotype has been advocated to have an important role in the fate of enteric pathogens residing in lettuce foliage. This study was therefore undertaken under the premise that different pathogen responses could occur in lettuce cultivars with cultivar selection being one of several hurdles in an overall strategy for controlling foodborne pathogens on field-grown produce. Up to eight lettuce cultivars ('Gabriella', 'Green Star', 'Muir', 'New Red Fire', 'Coastal Star', 'Starfighter', 'Tropicana', and 'Two Star') were examined in these experiments in which the plants were subjected to spray contamination of their foliage with pathogens. In an experiment that addressed internalization of Salmonella, cultivar was determined to be a significant variable (P < 0.05) with 'Gabriella' and 'Muir' being the least and most likely to exhibit internalization of this pathogen, respectively. Furthermore, antimicrobials (total phenols and antioxidant capacity chemicals) could be part of the plant's defenses to resist internalization as there was an inverse relationship between the prevalence of internalization at 1 h and the levels of these antimicrobials (r = -0.75 to -0.80, P = 0.0312 to 0.0165). Internalized cells appeared to be transient residents in that across all cultivars, plants sampled 1 h after being sprayed were 3.5 times more likely to be positive for Salmonella than plants analyzed 24 h after spraying (95% CI from 1.5 to 8.2, P = 0.0035). The fate of surface-resident Salmonella and Escherichia coli O157:H7 was addressed in subsequent growth chamber and field experiments. In the growth chamber study, no effect of cultivar was manifested on the fate of either pathogen when plants were sampled up to 12 days after spray contamination of their foliage. However, in the field study, five days after spraying the plants, Salmonella contamination was significantly affected by cultivar (P < 0.05) and the following order of prevalence of contamination was observed: 'Muir' < 'Gabriella' < 'Green Star' = 'New Red Fire' < 'Coastal Star'. Nine days after spray contamination of plants in the field, no effect of cultivar was exhibited due primarily to the low prevalence of contamination observed for Salmonella (8 of 300 plant samples positive by enrichment culture) and E. coli O157 (4 of 300 plant samples positive by enrichment culture). Given the narrow window of time during which cultivar differences were documented, it is unlikely that cultivar selection could serve as a viable option for reducing the microbiological risk associated with lettuce., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Cucumbers are frequently consumed raw and have been implicated in several recent foodborne outbreaks. Because this item may become contaminated at the farm, it is vital to explore the fate of attenuated Salmonella Typhimurium or Escherichia coli O157:H7 sprayed onto foliage, flowers, and fruit in fields and determine whether pre- or postcontamination spray interventions could minimize contamination. After spraying cucumber plants with contaminated irrigation water (3.8 log CFU/mL of Salmonella Typhimurium and E. coli O157:H7), 60 to 78% of cucumber fruit were not contaminated because the plant's canopy likely prevented many of the underlying fruit from being exposed to the water. Subsequent exposure of contaminated cucumber plants to a simulated shower event did not appear to dislodge pathogens from contaminated foliage onto the fruit, nor did it appear to consistently wash either pathogen from the fruit. Spraying flowers and attached ovaries directly with a pathogen inoculum (4.6 log CFU/mL) initially led to 100% and 65 to 90% contamination, respectively. Within 3 days, 30 to 40% of the flowers were still contaminated; however, contamination of ovaries was minimal (≤10%), suggesting it was unlikely that internalization occurred through the flower to the ovary with these pathogen strains. In another study, both pathogens were found on a withered flower but not on the fruit to which the flower was attached, suggesting that this contaminated flower could serve as a source of cross-contamination in a storage bin if harvested with the fruit. Because pre- and postcontamination acetic acid-based spray treatments failed to reduce pathogen prevalence, the probability that fruit initially contaminated at 1.3 to 2.8 log CFU of Salmonella Typhimurium or E. coli O157:H7 per cucumber would be positive by enrichment culture decreased by a factor of 1.6 and 1.9 for Salmonella Typhimurium and E. coli O157:H7, respectively, for every day the fruit was held in the field ( P ≤ 0.0001). Hence, to reduce the prevalence of Salmonella Typhimurium on cucumbers below 5%, more than 1 week would be required.

Compost is organic material that has been degraded into a nutrient-stabilized humus-like substance through intense microbial activity, which can provide essential plant nutrients (nitrogen, phosphorus) to aid in the growth of fruits and vegetables. Compost can be generated from animal waste feedstocks; these can contain human pathogens, which can be inactivated through the heat and microbial competition promoted during the composting process. Outbreaks of infections caused by bacterial pathogens such as Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on fruit and vegetable commodities consumed raw emphasize the importance of minimizing the risk of pathogenic contamination on produce commodities. This review article investigates factors that affect the reduction and survival of bacterial foodborne pathogens during the composting process. Interactions with indigenous microorganisms, carbon:nitrogen ratios, and temperature changes influence pathogen survival, growth, and persistence in finished compost. Understanding the mechanisms of pathogen survival during the composting process and mechanisms that reduce pathogen populations can minimize the risk of pathogen contamination in the cultivation of fruits and vegetables.

Field workers often wear gloves harvesting ready-to-eat produce; however, fields are not sterile environments and gloves may become contaminated numerous times during a working shift. This study explored the potential for inactivation of Escherichia coli O157:H7 and Salmonella when contaminated gloves were washed in levulinic acid (LV) and sodium dodecyl sulfate (SDS) solutions. Washing nitrile gloves with increasing concentrations of LV above 1.0% led to a decreased prevalence of glove contamination by Salmonella (P = 0.0000). A higher level of prevalence occurred for solid agar-cultured pathogens than liquid broth-cultured pathogens after nitrile gloves were washed in LV/SDS (P = 0.0000). Pathogens residing on latex gloves were more likely to be completely inactivated by washing in 0.5% LV/0.1% SDS solutions than nitrile or Canners gloves that exhibited inconsistent responses dependent on the pathogen strain. However, drying after washing nitrile gloves in 0.5% LV/0.1% SDS led to additional pathogen inactivation (P = 0.0394). Pathogen transfer from gloves to produce was implied as the pathogen prevalence on cantaloupe rind handled by LV/SDS-washed gloves was not statistically different from the prevalence on gloves (P = 0.7141). Hence, the risk of produce contamination may still exist but would be reduced by washing gloves in LV/SDS., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Gloves are worn by workers harvesting ready-to-eat produce as a deterrent for contaminating the produce with enteric pathogens that may reside on their hands. As fields are not sterile environments, the probability for gloves to become contaminated still exists and therefore it is critical to understand the conditions that affect the survival of pathogens on gloves. Both Escherichia coli O157:H7 and Salmonella deposited on glove surfaces in a liquid state survived longer when the pathogen had been suspended in lettuce sap than when suspended in water. Despite this protection, pathogens deposited on clean single-use gloves were more likely to survive during drying than pathogens deposited on dirty gloves (a film of lettuce sap had been applied to the surface prior to pathogen application and soil had been ground into the gloves). Survival of both E. coli O157:H7 and Salmonella was biphasic with the greatest losses occurring during the first hour of drying followed by much slower losses in the ensuing hours. Pathogens grown in rich media (tryptic soy broth) versus minimal media (M9) as well as those cultured on solid agar versus liquid broth were also more likely to be resistant to desiccation when deposited onto gloves. Although survival of E. coli O157:H7 on nitrile gloves was in general greater than it was on latex gloves, the relative survival of Salmonella on the two glove types was inconsistent. Due to these inconsistencies, no one glove type is considered better than another in reducing the risk for contamination with enteric pathogens. In addition, the extended survival of what are generally referred to as stress-resistant pathogens suggests that gloves either be changed frequently during the day or washed in a disinfectant to reduce the risk of glove contamination that could otherwise contaminate product handled with the contaminated gloves., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Consumers' criteria for evaluating food safety have evolved recently from considering the food's potential to cause immediate physical harm to considering the potential long-term effects that consumption of artificial ingredients, including antimicrobial preservatives, would have on health. As bacteriostatic and bactericidal agents to prevent microbial spoilage, antimicrobials not only extend shelf life, but they also enhance the product's safety. Antimicrobials and their levels that may be used in foods are specified by regulatory agencies. This review addresses the safety of antimicrobials and the potential consequences of removing those that are chemically synthesized or replacing them with antimicrobials from so-called natural sources. Such changes can affect the microbiological safety and spoilage of food as well as reduce shelf life, increase wastage, and increase the occurrence of foodborne illnesses.

Poor hygiene and improper food preparation practices in consumers' homes have previously been demonstrated as contributing to foodborne diseases. To address potential cross-contamination by kitchen utensils in the home, a series of studies was conducted to determine the extent to which the use of a knife or grater on fresh produce would lead to the utensil's contamination with Escherichia coli O157:H7 or Salmonella enterica. When shredding inoculated carrots (ca. 5.3 log CFU/carrot), all graters became contaminated and the number of E. coli O157:H7 present on the utensil was significantly greater than Salmonella (p < 0.05). Contamination of knives after slicing inoculated produce (4.9-5.4 log CFU/produce item) could only be detected by enrichment culture. After slicing tomatoes, honeydew melons, strawberries, cucumbers, and cantaloupes, the average prevalence of knife contamination by the two pathogens was 43%, 17%, 15%, 7%, and 3%, respectively. No significant increase in the incidence or level of contamination occurred on the utensils when residues were present (p > 0.05); however, subsequent contamination of 7 produce items processed with the contaminated utensils did occur. These results highlight the necessity of proper sanitization of these utensils when used in preparation of raw produce., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Microbial spoilage of salmon occurs during extended refrigerated storage and is often accompanied by unpleasant aromas. When spoilage is detected, it is assumed that consumers will reject the product for consumption. Because sensory panels of trained individuals or consumers are expensive and labor intensive, identification of microbiological or chemical indicators to characterize the extent to which fish has spoiled is needed when experimental process and storage treatments are being evaluated. A consumer panel of 53 senior citizens (60 to 85 years of age) evaluated in duplicate raw salmon subjected to 10 storage conditions, and the fish quality was targeted to range from fresh to very spoiled. This population group was chosen because they would be expected to have a greater prevalence of olfactory impairments and higher odor thresholds than the general population; in turn, a shorter safety margin or time period between product rejection due to spoilage and the generation of Clostridium botulinum toxins would be likely. Low hedonic scores for aroma and overall acceptance (2 or 3 of 9), corresponding to "dislike very much" to "dislike moderately," did not equate with unwillingness to prepare the sample for consumption by up to seven panelists (13%) when the product was presumed to have already been purchased. Despite these outliers, significant models (P = 0.0000) were developed for the willingness of consumers to prepare the sample for consumption and the sample's aerobic and anaerobic microbiological populations and two volatile peaks with Kovats indices of 640 and 753. However, these models revealed that the levels of microbiological and chemical markers must be very high before some consumers would reject the sample; hence, spoilage detection by smell would likely not be an adequate safeguard against consuming salmon in which C. botulinum toxin had been generated.

Shelf life of fish packaged under modified atmosphere (MA) is extended, but within the United States, commercial application of MA with impermeable packaging films is restricted due to concerns that botulinum toxin production would precede spoilage when contaminated fish are held at abusive storage temperatures. Use of semipermeable packaging films has been advocated; however, previous studies are inconclusive in determining the oxygen transmission rate (OTR) of a film that is needed to achieve an acceptable margin of safety (i.e., toxin production occurs only after spoilage). This study was conducted to determine the influence of OTR (target OTRs of 3 to 15,000) on the development of spoilage volatiles and toxin in salmon inoculated with type E Clostridium botulinum and subjected to air, vacuum, or 75:25 CO2:N2 MA and storage temperatures of 4, 8, 12, or 16°C. The most dominant headspace volatile peak that was produced during spoilage of samples at 4, 8 or 12°C was a peak, having a Kovats retention index (KI) of 753, and at which external standards of 2- or 3-methyl 1-butanol also eluted. Under anaerobic conditions, both the aerobic microbial populations and the size of the KI 753 spoilage peak were less in inoculated samples compared with uninoculated samples. C. botulinum-inoculated samples that were stored at 12 or 16°C under conditions favorable for anaerobic growth were also characterized by a KI 688 peak. Using a previously developed model that related the percentage of elderly consumers who would prepare a sample having the KI 753 spoilage peak of a specific size, it was determined that for salmon packaged with 3 or 3,000 OTR films under any atmosphere and stored at 12 or 16°C, 2 to 61% of the consumers could potentially prepare toxin-contaminated samples. Hence, when abusive storage conditions are suspected, the fish should not be consumed.

Consumers are being advised to increase their consumption of fruits and vegetables to reduce their risk of chronic disease. However, to achieve that goal, consumers must be able to implement protocols in their kitchens to reduce their risk of consuming contaminated produce. To address this issue, a study was conducted to monitor the fate of Escherichia coli O157:H7 and Salmonella on produce (cantaloupe, honeydew melon, carrots, and celery) that were subjected to brushing or peeling using common kitchen utensils. Removal of similar levels of Salmonella from carrots was accomplished by peeling and by brushing, but significantly greater removal of E. coli O157:H7 from carrots was accomplished by peeling than by brushing under running water (P < 0.05). Brushing removed significantly fewer pathogens from contaminated cantaloupes than from other produce items (P < 0.05), suggesting that the netted rind provided sites where the pathogen cells could evade the brush bristles. A Sparta polyester brush was less effective than a scouring pad for removing Salmonella from carrots (P < 0.05). In all cases, brushing and peeling failed to eliminate the pathogens from the produce items, which may be the result of contamination of the utensil during use. High incidences of contamination (77 to 92%) were found among peelers used on carrots or celery, the Sparta brush used on carrots, and the scouring pad used on carrots and cantaloupe. Of the utensils investigated, the nylon brush had the lowest incidence of pathogen transference from contaminated produce (0 to 12%). Transfer of pathogens from a potentially contaminated Sparta brush or peeler to uncontaminated carrots did not occur or occurred only on the first of seven carrots processed with the utensil. Therefore, risk of cross-contamination from contaminated utensils to uncontaminated produce may be limited.

Freshly harvested Eastern variety cantaloupes (Cucumis melo L. var. reticulatus cv. Athena) were subjected to three different harvest and wash treatments to examine conditions under which the efficacy of the sanitizer, levulinic acid (LV) plus sodium dodecyl sulfate (SDS), could be enhanced to reduce Salmonella contamination. In treatment set one, cantaloupes were spot inoculated with Salmonella enterica serovar Poona (prepared from solid or liquid media cultures) before or after a 1-min dip treatment in LV (2.5, 5.0, 7.5, or 10%) and 2.5% SDS. S. Poona initial populations on rind tissue (4.26-5.04 log CFU/sample) were reduced to detection by enrichment culture when cantaloupes were subsequently exposed to any of the LV/SDS solutions. When S. Poona was introduced after cantaloupes had been dip-treated, greater decreases in pathogen populations at the stem scar were observed when cantaloupes were treated with increasing concentrations of LV. In treatment set two, the response of S. Poona dip-treated with 5% LV/2.5% SDS was compared to a simulated commercial dump tank treatment incorporating 200 ppm chlorine as well as a two-stage treatment employing both the chlorine tank and LV/SDS dip treatments. S. Poona levels (log CFU/sample or # positive by enrichment culture/# analyzed) after treatments were 5.25, 3.07, 7/10, 5/10 (stem scar) and 3.90, 25/40, 28/40, 20/40 (rind) for non-treated, chlorine tank, LV/SDS dip, and tank plus dip treatments, respectively. In treatment set three, freshly harvested cantaloupes were first treated in the field using a needle-free stem scar injection (200 μl, 7.5% LV/1.0% SDS, 60 psi) and a cantaloupe spray (30 ml, 7.5% LV/0.5% SDS). Cantaloupe stem scar and rind tissue were then spot-inoculated with S. Poona using either a liquid or soil-based medium followed by a simulated dump tank treatment incorporating either 200 ppm chlorine or 5% LV/2% SDS. S. Poona inoculated on field-treated cantaloupe rind decreased by 4.7 and 5.31 (liquid) and 3.27 and 3.36 (soil) log CFU/sample after simulated chlorine and LV/SDS tank treatments, respectively. In the case of stem scar tissue, S. Poona populations exhibited a 1.0 log greater reduction when cantaloupes were treated with LV/SDS compared to chlorine in the dump tank (P<0.05). Based on this study, application of multiple hurdles is warranted, as additional decreases in S. Poona populations were obtained when cantaloupes were subjected to a chlorine dump tank followed by a LV/SDS dip treatment., (Copyright © 2015 Elsevier B.V. All rights reserved.)

During the past century, the microbiological safety of the US food supply has improved; however, many foodborne illnesses and outbreaks occur annually. Hence, opportunities for the food industry to improve the safety of both domestic and imported food exist through the adoption of risk-based preventive measures. Challenging food safety issues that are on the horizon include demographic changes to a population whose immune system is more susceptible to foodborne and opportunistic pathogens, climate changes that will shift where food is produced, and consumers' preferences for raw and minimally processed foods. Increased environmental and product testing and anonymous data sharing by the food industry with the public health community would aid in identifying system weaknesses and enabling more targeted corrective and preventive actions. Clinicians will continue to play a major role in reducing foodborne illnesses by diagnosing and reporting cases and in helping to educate the consumer about food safety practices., (© The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Heat is the primary mechanism by which aerobic composting inactivates zoonotic bacterial pathogens residing within animal manures, but at sublethal temperatures, the time necessary to hold the compost materials to ensure pathogen inactivation is uncertain. To determine the influence of the type of nitrogen amendment on inactivation of Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 in compost mixtures stored at sublethal temperatures, specific variables investigated in these studies included the animal source of the manure, the initial carbon/nitrogen (C:N) ratio of the compost mixture, and the age of the manure. Salmonella and L. monocytogenes were both inactivated more rapidly in chicken and swine compost mixtures stored at 20°C when formulated to an initial C:N ratio of 20:1 compared with 40:1, whereas a C:N ratio did not have an effect on inactivation of these pathogens in cow compost mixtures. Pathogen inactivation was related to the elevated pH of the samples that likely arises from ammonia produced by the indigenous microflora in the compost mixtures. Indigenous microbial activity was reduced when compost mixtures were stored at 30°C and drier conditions (<10% moisture level) were prevalent. Furthermore, under these drier conditions, Salmonella persisted to a greater extent than L. monocytogenes, and the desiccation resistance of Salmonella appeared to convey cross-protection to ammonia. Salmonella persisted longer in compost mixtures prepared with aged chicken litter compared with fresh chicken litter, whereas E. coli O157:H7 survived to similar extents in compost mixtures prepared with either fresh or aged cow manure. The different responses observed when different sources of manure were used in compost mixtures reveal that guidelines with times required for pathogen inactivation in compost mixtures stored at sublethal temperatures should be dependent on the source of nitrogen, i.e., type of animal manure, present.

During the early phases of aerobic composting of animal manures, pathogens are inactivated primarily from the accumulation of heat produced by indigenous microbial activity. When compost materials are not exposed to these lethal temperatures, the required holding time needed to obtain a pathogen-free product that may be applied to fields is unknown. Consequently, a series of studies examined whether the carbon amendment (wheat straw, peanut hulls, rice hulls, and pine needles) added to animal manures affected survival of either Salmonella or E. coli O157:H7 during storage of compost mixtures at sublethal temperatures (20 to 40°C). Pathogens consistently survived for longer periods of time in compost mixtures prepared with pine needles than compost mixtures prepared with either of the other three carbon amendments. Pathogen inactivation in wheat straw- or peanut hull-amended compost mixtures was dependent on the target pathogen, moisture level, and storage temperature. Moisture levels in wheat straw-amended compost mixtures stored at 40°C had no effect on inactivation of E. coli O157:H7. In contrast, wheat straw-amended mixtures stored at 30 to 35°C and equilibrated to suboptimal moisture contents (30 to 40%) were less effective for inactivating pathogens compared with drier (25% moisture) or moister (60% moisture) mixtures. In peanut hull-amended compost mixtures, inactivation of E. coli O157:H7 was affected minimally by moisture levels, whereas Salmonella survival increased as the moisture level was decreased. The different inactivation responses of Salmonella and E. coli O157:H7 in compost mixtures prepared with wheat straw or peanut hulls and equilibrated to different moisture levels suggest that there are different mechanisms for inactivation. Hence, developing reliable guidelines relying on time-temperature for holding of compost mixtures at sublethal temperatures will be challenging and, perhaps, not possible.

Two separate studies were conducted to address the condition and the type of feedstocks used during composting of dairy manure. In each study, physical (temperature), chemical (ammonia, volatile acids, and pH), and biological (Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7) parameters were monitored during composting in bioreactors to assess the degree to which they were affected by the experimental variables and, ultimately, the ability of the chemical and physical parameters to predict the fate of pathogens during composting. Compost mixtures that contained either aged dairy manure or pine needles had reduced heat generation; therefore, pathogen reduction took longer than if fresh manure or carbon amendments of wheat straw or peanut hulls were used. Based on regression models derived from these results, ammonia concentration, in addition to heat, were the primary factors affecting the degree of pathogen inactivation in compost mixtures formulated to an initial carbon-nitrogen (C:N) ratio of 40:1, whereas, the pH of the compost mixture along with the amount of heat exposure were most influential in compost mixtures formulated to an initial C:N ratio of 30:1. Further studies are needed to validate these models so that additional criteria in addition to time and temperature can be used to evaluate the microbiological safety of composted manures.

Escherichia coli O157:H7 has been the causative agent of many outbreaks associated with leafy green produce consumption. Elucidating the mechanism by which contamination occurs requires monitoring interactions between the pathogen and the plant under typical production conditions. Intentional introduction of virulent strains into fields is not an acceptable practice. As an alternative, attenuated strains of natural isolates have been used as surrogates of the virulent strains; however, the attachment properties and environmental stabilities of these attenuated isolates may differ from the unattenuated outbreak strains. In this study, the Shiga toxin (stx1, stx2, and/or stx2c) genes as well as the eae gene encoding intimin of two E. coli O157:H7 outbreak isolates, F4546 (1997 alfalfa sprout) and K4492 (2006 lettuce), were deleted. Individual gene deletions were confirmed by polymerase chain reaction (PCR) and DNA sequencing. The mutant strains did not produce Shiga toxin. The growth kinetics of these mutant strains under nutrient-rich and minimal conditions were identical to those of their wild-type strains. Attachment to the surface of lettuce leaves was comparable between wild-type/mutant pairs F4546/MD46 and K4492/MD47. Adherence to soil particles was also comparable between the virulent and surrogate pairs, although the F4546/MD46 pair exhibited statistically greater attachment than the K4492/MD47 pair (p≤0.05). Wild-type and mutant pairs F4546/MD46 and K4492/MD47 inoculated into wet or dry soils had statistically similar survival rates over the 7-day storage period at 20°C. A plasmid, pGFPuv, containing green fluorescent protein was transformed into each of the mutant strains, allowing for ease of identification and detection of surrogate strains on plant material or soil. These pGFPuv-containing surrogate strains will enable the investigation of pathogen interaction with plants and soil in the farm production environment where the virulent pathogen cannot be used.

Reduction of enteric pathogens in animal manures before field application is essential for mitigating the risk of foodborne illness associated with produce. Aerobic composting of manures has been advocated as an effective treatment for reducing pathogen populations, and heat is a major factor contributing to pathogen inactivation. This study was initiated to determine the potential contribution of both thermal and nonthermal (pH, volatile acids, and ammonia) factors to pathogen inactivation during aerobic composting in bioreactors for mixtures containing manure from various sources (dairy, chicken, and swine). The test mixtures were formulated with an initial moisture content of 60% and a C:N ratio of 20:1, using straw and cottonseed meal as amendments. Mixtures were then inoculated with Salmonella and Listeria monocytogenes labeled with green fluorescent protein at initial populations of ca. 10(7) CFU/g. Three replicate trials of each treatment were conducted. Temperatures within the bioreactors were recorded at 30-min intervals, and duplicate samples were withdrawn daily from two sampling locations within the bioreactor. Significant regression models were derived relating decreases in pathogen populations to the degree of heat generated in the mixture (cumulative heat) and the pH of the mixture on the day before the pathogen losses were calculated (P < 0.0002). Although pathogens in swine manure compost mixtures were inactivated by the third day of composting, very little heat was generated in these mixtures, which were characterized by significantly higher levels of volatile acids compared with the other two compost mixtures. Therefore, volatile acids could help achieve pathogen inactivation when temperatures are too low such as when heat is lost too quickly at the surface of static compost piles or during winter composting.

Preharvest internalization of Escherichia coli O157:H7 into the roots of leafy greens is a food safety risk because the pathogen may be systemically transported to edible portions of the plant. In this study, both abiotic (degree of soil moisture) and biotic (E. coli O157:H7 exposure, presence of Shiga toxin genes, and type of leafy green) factors were examined to determine their potential effects on pathogen internalization into roots of leafy greens. Using field soil that should have an active indigenous microbial community, internalized populations in lettuce roots were 0.8 to 1.6 log CFU/g after exposure to soil containing E. coli O157:H7 at 5.6 to 6.1 log CFU/g. Internalization of E. coli O157:H7 into leafy green plant roots was higher when E. coli O157:H7 populations in soil were increased to 7 or 8 log CFU/g or when the soil was saturated with water. No differences were noted in the extent to which internalization of E. coli O157:H7 occurred in spinach, lettuce, or parsley roots; however, in saturated soil, maximum levels in parsley occurred later than did those in spinach or lettuce. Translocation of E. coli O157:H7 from roots to leaves was rare; therefore, decreases observed in root populations over time were likely the result of inactivation within the plant tissue. Shiga toxin-negative (nontoxigenic) E. coli O157:H7 isolates were more stable than were virulent isolates in soil, but the degree of internalization of E. coli O157:H7 into roots did not differ between isolate type. Therefore, these nontoxigenic isolates could be used as surrogates for virulent isolates in field trials involving internalization.

In the past decade, leafy greens have been implicated in several outbreaks of foodborne illness, and research has focused on contamination during preharvest operations. Concerns have been raised that internalization of pathogens into the edible tissue occurs where postharvest chemical interventions would be ineffective. This study was initiated to measure the degree and fate of Escherichia coli O157:H7 internalized in the phyllosphere tissue of leafy greens when spray conditions, inoculum level, and type of leafy green were varied. Two spraying treatments were applied: (i) spraying individual spinach or lettuce leaves on plants once with a high dose (7 to 8 log CFU/ml) of E. coli O157:H7 and (ii) spraying spinach, lettuce, or parsley plants repeatedly (once per minute) with a low dose (2.7 to 4.2 log CFU/ml) of E. coli O157:H7 over a 10- to 20-min period. With the high-dose spray protocol, no significant differences in the prevalence of internalization occurred between Shiga toxin-negative E. coli O157:H7 isolates and virulent isolates (P > 0.05), implying that the Shiga toxin virulence factors did not influence internalization or the subsequent fate of those populations under these test conditions. Significantly greater internalization of E. coli O157:H7 occurred in spinach leaves compared with lettuce leaves when leaves were sprayed once with the high-dose inoculum (P < 0.05), whereas internalization was not observed in lettuce leaves but continued to be observed in spinach and parsley leaves following repeated spraying of the low-dose inoculum. Based on these results, it is surmised that a moisture film was generated when spraying was repeated and this film assisted in the mobilization of pathogen cells to plant apertures, such as stomata. E. coli O157:H7 cells that were internalized into spinach tissue using a low-dose repeat-spray protocol were temporary residents because they were not detected 2 days later, suggesting that plant-microbe interactions may be responsible.

Aims: Three soils that varied in their physicochemical characteristics and microbial diversity were inoculated with Escherichia coli O157:H7 and Salmonella to determine the relative impact of abiotic and biotic factors on the pathogens' survival when the soil was held at 25°C., Methods and Results: Three soils that were classified as having low, medium and high microbial diversity were divided into two batches for adjustment to 20% of water-holding capacity and to 40% of water-holding capacity. Soils were inoculated with both green fluorescent-labelled E. coli O157:H7 and red fluorescent-labelled Salmonella (5 log CFU g(-1) dry weight) and held at 25°C. Pathogens inoculated into an acidic soil died off within 9 weeks, whereas they were still detected in the other two soils by enrichment culture after 18 weeks. Moisture did not affect inactivation of E. coli O157:H7, but did affect Salmonella inactivation in soil having the greatest organic load and microbial diversity. Using multiple linear regression analysis, 98.7% of the variability in the inactivation rate for E. coli O157:H7 was explained by a model that included the variables of initial pH and electrical conductivity. Salmonella's inactivation rate was predicted by a model that included pH and initial cell numbers of copiotrophic and oligotrophic bacteria., Conclusion: This study provided evidence of specific properties that impact inactivation of E. coli O157:H7 and Salmonella in soils at 25°C., Significance and Impact of the Study: Identification of factors influential in the die-off of enteric pathogens will assist in developing guidelines for safe intervals between field contamination events and planting or harvesting of fresh-cut produce crops., (© 2013 The Society for Applied Microbiology.)

Both growth chamber and field studies were conducted to investigate the potential for Escherichia coli O157:H7 to be internalized into leafy green tissue when seeds were germinated in contaminated soil. Internalized E. coli O157:H7 was detected by enrichment in both spinach (Spinacia oleracea L.) and lettuce (Lactuca sativa L.) seedlings when seeds were germinated within the growth chamber in autoclaved and nonautoclaved soil, respectively, contaminated with E. coli O157:H7 at 2.0 and 3.8 log CFU/g, respectively. Internalized E. coli O157:H7 populations could be detected by enumeration within leafy green tissues either by increasing the pathogen levels in the soil or by autoclaving the soil. Attempts to maximize the exposure of seed to E. coli O157:H7 by increasing the mobility of the microbe either through soil with a higher moisture content or through directly soaking the seeds in an E. coli O157:H7 inoculum did not increase the degree of internalization. Based on responses obtained in growth chamber studies, internalization of E. coli O157:H7 surrogates (natural isolates of Shiga toxin-negative E. coli O157:H7 or recombinant [stx- and eae-negative] outbreak strains of E. coli O157:H7) occurred to a slightly lesser degree than did internalization of the virulent outbreak strains of E. coli O157:H7. The apparent lack of internalized E. coli O157:H7 when spinach and lettuce were germinated from seed in contaminated soil (ca. 3 to 5 log CFU/g) in the field and the limited occurrence of surface contamination on the seedlings suggest that competition from indigenous soil bacteria and environmental stresses were greater in the field than in the growth chamber. On the rare occasion that soil contamination with E. coli O157:H7 exceeded 5 log CFU/g in a commercial field, this pathogen probably would not be internalized into germinating leafy greens and/or would not still be present at the time of harvest.

Background: The survival and distribution of enteric pathogens in soil and lettuce systems were investigated in response to several practices (soil amendment supplementation and reduced watering) that could be applied by home gardeners., Results: Leaf lettuce was grown in manure compost/top soil (0:5, 1:5 or 2:5 w/w) mixtures. Escherichia coli O157:H7 or Salmonella was applied at a low or high dose (10(3) or 10(6) colony-forming units (CFU) mL(-1) ) to the soil of seedlings and mid-age plants. Supplementation of top soil with compost did not affect pathogen survival in the soil or on root surfaces, suggesting that nutrients were not a limiting factor. Salmonella populations on root surfaces were 0.7-0.8 log CFU g(-1) lower for mid-age plants compared with seedlings. E. coli O157:H7 populations on root surfaces were 0.8 log CFU g(-1) lower for mid-age plants receiving 40 mL of water compared with plants receiving 75 mL of water on alternate days. Preharvest internalization of E. coli O157:H7 and Salmonella into lettuce roots was not observed at any time., Conclusion: Based on the environmental conditions and high pathogen populations in soil used in this study, internalization of Salmonella or E. coli O157:H7 into lettuce roots did not occur under practices that could be encountered by inexperienced home gardeners., (© 2013 Society of Chemical Industry.)

Both spinach and lettuce were grown to harvest, cut, and then regrown after spraying the cut shoots with irrigation water contaminated with Escherichia coli O157:H7. Plant tissue was collected on the day of spraying and again 2 and 14 days later for analysis of total and internalized E. coli O157:H7 populations. Internalization of E. coli O157:H7 occurred on the day of spraying, and larger populations were internalized as the level in the spray increased. Tissue repair was slow and insufficient to prevent infiltration of E. coli O157:H7; internalized E. coli O157:H7 in shoots cut 5 days prior to exposure to E. coli O157:H7-contaminated water were not significantly different from levels in shoots cut on the same day of spraying with contaminated water (P > 0.05). Two days after spraying plants with a high level of E. coli O157:H7 (7.3 log CFU/ml), levels of internalized E. coli O157:H7 decreased by ca. 2.6 and 1.3 log CFU/g in Tyee and Bordeaux spinach, respectively, whereas populations of internalized E. coli O157:H7 decreased very little (ca. 0.4 log CFU/g) in lettuce plants that had been sprayed either on the same day as cutting or 1 day after cutting. When cut plants were sprayed with irrigation water at a lower contamination level (4.5 log CFU/ml), internalized E. coli O157:H7 was not detected in either spinach or lettuce plants 2 days later and therefore would not likely be of concern when the crop was harvested.

Freshly harvested Georgia-grown cantaloupes (Cucumis melo L. var. reticulatus cv. Athena and Atlantis) were spot inoculated with 100 μl of a five-strain mixture of Salmonella enterica serovar Poona (9 log CFU/ml) at the stem scar and on the netted rind and then subjected to no treatment (control) or a 6-min treatment (tank only) in water, 120 ppm of chlorine (pH 7.0), 1% levulinic acid plus 0.1% sodium dodecyl sulfate (SDS; pH 3.0), or 2% levulinic acid plus 0.2% SDS (pH 3.0). The log reduction for the tank-only treatments was 0.31, 0.59, 1.32, and 1.37 log CFU/g at the stem scar and 0.97, 1.59, 2.06. and 3.37 log CFU/g on the netted rind for water, chlorine, 1% levulinic acid plus 0.1% SDS, and 2% levulinic acid plus 0.2% SDS, respectively. A greater log reduction was observed for the cantaloupe surface tissue with the water, chlorine, and 2% levulinic acid plus 0.2% SDS treatments when additional sanitizer (2 ml) and brushing (to simulate cantaloupes tumbling over brushes on the processing line) were added to the dump tank treatment. The stem scar tissue reductions were 0.90, 1.69, and 1.53 log CFU/g, whereas the netted rind reductions were 1.56, 2.50, and 4.47 log CFU/g after treatment with water, chlorine, and 2% levulinic acid plus 0.2% SDS, respectively. These data suggest that 2% levulinic acid plus 0.2% SDS is effective for reducing Salmonella on the netted rind surface of cantaloupes. However, neither 2% levulinic acid plus 0.2% SDS nor 120 ppm of chlorine substantially reduced Salmonella on stem scar tissue.

Human noroviruses and hepatitis A virus are responsible for numerous outbreaks associated with handling fresh produce. In this study, physical removal of hepatitis A virus and murine norovirus, a human norovirus surrogate, from contaminated produce items (honeydew melons, cantaloupes, carrots, and celery) by scrubbing under running water with a nylon brush or scouring pad and by peeling (carrots and celery) with a peeler was investigated. The degree and extent of utensil contamination with viruses during these operations in the presence and absence of food residue also was investigated. Scrubbing or peeling produce initially inoculated with ∼5.5 log PFU of each virus resulted in significant levels of virus removal, ranging from 0.93 to 2.85 log PFU. However, utensil cross-contamination occurred, with >2 log PFU of virus transferred from a single produce item. After preparation of a contaminated produce item, utensil cross-contamination resulted in virus detection on seven successively prepared produce items. Produce residue accumulation on utensils variably impacted virus transfer to utensil surfaces. Results indicate that scrubbing and peeling produce can reduce levels of viruses on contaminated produce, but the importance of utensil sanitation to prevent cross-contamination is highlighted. Findings also provide important information for modeling virus cross-contamination during food preparation.

Fruits, vegetables, and meat are susceptible to contamination by foodborne pathogens at many points from production through preparation in the home. This review will largely highlight approaches and progress made in the last five years to address strategies to reduce pathogen contamination in animal production but will also touch on the emerging field of preharvest produce food safety. Mitigation strategies can be divided into those that address pathogen reduction in the environment and those that target reduction/elimination of pathogen contamination in animals or plants. The former strategy has been encompassed in studies evaluating sanitation treatments of facilities as well as in numerous epidemiologic risk assessment studies (both on-farm assessments and computer simulation models) that identify management practices that impact pathogen prevalence in animals. Interventions to significantly reduce pathogen exposure via feed or water are dependent on their role as a significant contributor to pathogen contamination in the animal production system. In addition, inconsistent results obtained with interventions of dietary additives or formulation modifications (grain versus forage; inclusion of distiller's grains) on pathogen prevalence in animals have been attributed to a range of factors including target organism, grain type, level of inclusion, the animal's health or stress level, and ability to survive the gastric acidic conditions. Recent attempts to microencapsulate organic acids or bacteriophage within feed have met with only marginal improvements in reducing pathogen carriage in animals but this approach may have greater potential with other antimicrobial additives (i.e., essential oils). Bacteriophage therapy, in general, can significantly reduce pathogen carriage in animals but based on its transient nature and the potential for development of phage-resistant subpopulations, this approach should be administered to animals just prior to slaughter and preferably to animals that are suspected "super-shedders". Other promising on-farm intervention approaches have included breeding for pathogen resistance, vaccines, and dietary bacteriocins. To optimize interventions on a cost basis, studies have also determined that application of dietary interventions at specific time points in the animal's production cycle is a useful strategy to reduce pathogen carriage (e.g., probiotics to fertilized eggs and acidified feed to fattening swine). In conclusion, applicable management and intervention strategies may vary depending on the type of food under production; however, it is important to consider from a holistic view how any new intervention strategies will affect the overall production system in order to maintain a successful, efficient food production environment., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Recent studies addressing the internalization of fresh produce by foodborne pathogens arose in response to the growing number of recent and high profile outbreaks involving fresh produce. Because chemical sanitizing agents used during harvest and minimal processing are unlikely to reach enteric pathogens residing within plant tissue, it is imperative that paths for pathogen entry be recognized and minimized. Using both microscopy and microbial enumeration tools, enteric pathogens have been shown to enter plant tissues through both natural apertures (stomata, lateral junctions of roots, flowers) and damaged (wounds, cut surfaces) tissue. In studies revealing preharvest internalization via plant roots or leaf stomata, experimental conditions have primarily involved exposure of plants to high pathogen concentrations (≥ 6 log g⁻¹ soil or 6 log ml⁻¹ water), but those pathogens internalized appear to have short-term persistence. Postharvest internalization of pathogens via cut surfaces may be minimized by maintaining effective levels of sanitizing agents in waters during harvesting and minimal processing.

Throughout four field trials, compost heaps covered with finished compost maintained temperatures under the physical covering that were ca. 7 to 15.5°C higher, resulting in rapid Escherichia coli O157:H7 reduction, than those of the heaps covered with fresh straw or left uncovered. Our results validated recommendations made by the U.S. Environmental Protection Agency for covering fresh compost.

The microbiological safety of cabbage, carrots, celery, and onions/scallions as well as deli (mayonnaise-based) salads that contain these items is the subject of this review. Between 2000 and 2007, the number of outbreaks in the United States associated with these raw produce items ranged from 6 (celery) to 18 (carrots). For cases with confirmed etiologies involving these 4 types of produce as well as coleslaw, chicken, seafood, and other vegetable-based salads, more than 50% of the outbreaks were attributed to viral agents. In contrast, Salmonella spp. served as the major etiological agent in outbreaks associated with potato salad. Surveys conducted on these produce items within the United States and other developed countries found either an absence or infrequent contamination with foodborne pathogens. Despite this low prevalence, experimental studies have demonstrated the potential for preharvest contamination, and this event is more likely to occur when exposure is close to harvest. Postharvest contamination of these produce items has been documented in several cases with water, equipment, and incoming product serving as the principal cross-contamination agent. Survival of contaminated product during subsequent storage is dependent on the storage temperature, produce type, and presence of mayonnaise. Chemical interventions may be relied on to reduce cross-contamination during produce washing operations but are limited in their ability to inactivate pathogens on the produce surface. In contrast, irradiation at dosages (1.0 kGy) approved for use in the United States is an effective treatment for killing pathogenic bacteria in fresh-cut cabbage, carrots, and celery., (© 2010 Institute of Food Technologists®.)

Environmental pests may serve as reservoirs and vectors of zoonotic pathogens to leafy greens; however, it is unknown whether insect pests feeding on plant tissues could redistribute these pathogens present on the surface of leaves to internal sites. This study sought to differentiate the degree of tissue internalization of Escherichia coli O157:H7 when applied at different populations on the surface of lettuce and spinach leaves, and to ascertain whether lettuce-infesting insects or physical injury could influence the fate of either surface or internalized populations of this enteric pathogen. No internalization of E. coli O157:H7 occurred when lettuce leaves were inoculated with 4.4 log CFU per leaf, but it did occur when inoculated with 6.4 log CFU per leaf. Internalization was statistically greater when spinach leaves were inoculated on the abaxial (underside) than when inoculated on the adaxial (topside) side, and when the enteric pathogen was spread after surface inoculation. Brief exposure (∼18 h) of lettuce leaves to insects (5 cabbage loopers, 10 thrips, or 10 aphids) prior to inoculation with E. coli O157:H7 resulted in significantly reduced internalized populations of the pathogen within these leaves after approximately 2 weeks, as compared with leaves not exposed to insects. Surface-contaminated leaves physically injured through file abrasions also had significantly reduced populations of both total and internalized E. coli O157:H7 as compared with nonabraded leaves 2 weeks after pathogen exposure.

Bacteriophages specific to Salmonella strains were isolated from sewage effluent and characterized. A five-strain bacteriophage mixture was applied to dairy manure compost inoculated with Salmonella enterica serotype Typhimurium. Bacteriophage treatment resulted in a greater than 2-log-unit reduction of Salmonella within 4 h at all moisture levels compared to the controls.

Numerous field studies have revealed that irrigation water can contaminate the surface of plants; however, the occurrence of pathogen internalization is unclear. This study was conducted to determine the sites of Escherichia coli O157:H7 contamination and its survival when the bacteria were applied through spray irrigation water to either field-grown spinach or lettuce. To differentiate internalized and surface populations, leaves were treated with a surface disinfectant wash before the tissue was ground for analysis of E. coli O157:H7 by direct plate count or enrichment culture. Irrigation water containing E. coli O157:H7 at 10(2), 10(4), or 10(6) CFU/ml was applied to spinach 48 and 69 days after transplantation of seedlings into fields. E. coli O157:H7 was initially detected after application on the surface of plants dosed at 10(4) CFU/ml (4 of 20 samples) and both on the surface (17 of 20 samples) and internally (5 of 20 samples) of plants dosed at 10(6) CFU/ml. Seven days postspraying, all spinach leaves tested negative for surface or internal contamination. In a subsequent study, irrigation water containing E. coli O157:H7 at 10(8) CFU/ml was sprayed onto either the abaxial (lower) or adaxial (upper) side of leaves of field-grown lettuce under sunny or shaded conditions. E. coli O157:H7 was detectable on the leaf surface 27 days postspraying, but survival was higher on leaves sprayed on the abaxial side than on leaves sprayed on the adaxial side. Internalization of E. coli O157:H7 into lettuce leaves also occurred with greater persistence in leaves sprayed on the abaxial side (up to 14 days) than in leaves sprayed on the adaxial side (2 days).

Aims: The purpose of this study was to determine whether the methods used in compost operations of small and medium-sized poultry farms resulted in the production of an amendment free of foodborne pathogens., Methods and Results: Nine compost heaps on five South Carolina poultry farms were surveyed at different stages of the composting process. Compost samples were analysed for coliforms and enriched for Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes. The waste materials and composting practices differed among the surveyed farms. On two farms, new materials were added to heaps that had previously completed the active composting phase. Five compost heaps did not reach an internal temperature of 55 degrees C, and c. 62% of all internal samples in the first composting phase contained moisture contents <40%. Escherichia coli was detected in 63% of the surface samples (n = 38) and 9.8% of the internal samples (n = 82) from the first composting phase, as compared with 16.7% of the surface samples (n = 12) and 0% internal samples (n = 24) from the second composting phase. Salmonella was detected in 26 and 6.1% of all surface and internal samples collected from heaps in the first composting phase, respectively, but was absent in all compost samples undergoing a second composting phase. The predominant Salmonella serotypes were Thompson, Montevideo and Anatum. Neither E. coli O157:H7 nor L. monocytogenes was detected in any of the samples., Conclusions: Our results indicate that the conditions at the compost surface are suitable for pathogen survival, and the complete composting process can result in the elimination of pathogens in poultry wastes., Significance and Impact of the Study: This research provides information regarding the effectiveness of the composting practices and microbiological quality of poultry compost produced by small- and medium-sized farms. Ensuring the safety of compost that may be applied to soils should be an integral part of preharvest food safety programme.

Several sources of contamination of fresh produce by Escherichia coli O157:H7 (O157) have been identified and include contaminated irrigation water and improperly composted animal waste; however, field studies evaluating the potential for internalization of O157 into leafy greens from these sources have not been conducted. Irrigation water inoculated with green fluorescent plasmid-labeled Shiga toxin-negative strains (50 ml of 10(2), 10(4), or 10(6) CFU of O157 per ml) was applied to soil at the base of spinach plants of different maturities in one field trial. In a second trial, contaminated compost (1.8 kg of 10(3) or 10(5) CFU of O157 per g) was applied to field plots (0.25 by 3.0 m) prior to transplantation of spinach, lettuce, or parsley plants. E. coli O157:H7 persisted in the soil up to harvest (day 76 posttransplantation) following application of contaminated irrigation water; however, internalized O157 was not detected in any spinach leaves or in roots exposed to O157 during the early or late growing season. Internalized O157 was detected in root samples collected 7 days after plants were contaminated in mid-season, with 5 of 30 samples testing positive for O157 by enrichment; however, O157 was not detected by enrichment in surface-disinfected roots on days 14 or 22. Roots and leaves from transplanted spinach, lettuce, and parsley did not internalize O157 for up to 50 days in the second trial. These results indicate that internalization of O157 via plant roots in the field is rare and when it does occur, O157 does not persist 7 days later.

The fate of manure-borne pathogen surrogates (gfp-labeled Escherichia coli O157:H7 and Listeria innocua and avirulent Salmonella Typhimurium) in the field was monitored at both sub-surface (30 cm from surface) and surface sites of static composting piles (3.5-m base diameter) composed of chicken litter and peanut hulls. Despite exposure to elevated temperatures, Salmonella was detected by enrichment culture in sub-surface samples following 14 days of composting. In surface samples, pathogen surrogates were detected in the summer after 4 days of composting by enrichment culture only, whereas E. coli O157:H7 and L. innocua remained detectable by direct plating (>2 log(10)cfu/g) up to 28 days in piles composted during the fall and winter. All three types of bacteria remained detectable by enrichment culture in surface samples composted for 56 days during the winter.

Aerobic composting is a common management practice to inactivate pathogens in manure; however, additional research on the role of compost composition in pathogen inactivation is needed. The objective of this study was therefore to determine the effect of the carbon:nitrogen (C:N) ratio and the presence of ammonium sulfate on inactivation of Salmonella spp. in cow manure-based mixtures composted in a bioreactor under controlled conditions. Compost preparations with an initial C:N ratio of 20:1 required a maximum of 4 days of storage before Salmonellae were inactivated by 7 log(10), whereas preparations with C:N ratios of 30:1 and 40:1 C:N required more than 5 and 7 days of storage, respectively. The pH values of both the 20:1 and 30:1 C:N preparations decreased during the onset of composting before increasing to >8. In contrast, pH values of 40:1 C:N preparations increased immediately to >8, generally within the first day of storage. Maximum temperatures observed in 20:1 C:N preparations for inactivation of pathogens were less than 50 degrees C, and the cumulative heat exposure required for pathogen inactivation in 20:1 C:N preparations was 15-fold less than in 40:1 C:N preparations. Supplementation of compost mixtures with 0.08% ammonium sulfate resulted in slightly higher temperatures; however, these higher temperatures did not translate into more rapid rates of pathogen inactivation.

Studies were done to determine the effect of heat stress on internalization of Escherichia coli O157:H7 in lettuce subjected to different watering practices during growth. Iceberg and romaine lettuce were grown in sandy soil in an environmental chamber at 23 degrees C during the day and 7 degrees C at night, with a 12-h photoperiod. Thirty days after transplanting seedlings, potting soil was inoculated with a five-strain mixture of green fluorescent protein-labeled E. coli O157:H7 at populations of 4 and 6 log CFU/g of soil. Lettuce plants were exposed to one of two temperature stress regimes: 36 degrees C during the day and 15 degrees C at night for 2 days, or 32 degrees C during the day and 15 degrees C at night for 3 days, both with a 12-h photoperiod. Control plants were held at 23 degrees C during the day and 7 degrees C at night for 3 days. Plants were either watered daily or not watered during the heat stress and control treatments. E. coli O157:H7 was detected by enrichment in all inoculated soil and rhizosphere samples from plants grown in inoculated soil. Less E. coli O157:H7 was detected in inoculated heat-stressed soil than in control soil. From inoculated pots, all leaf surfaces and macerated leaves that had been surface sanitized were negative for E. coli O157:H7. All surface-sanitized macerated roots from control samples and from 143 of 144 samples of inoculated samples were negative for E. coli O157:H7. Heat stress during growth of lettuce did not promote or enhance internalization of E. coli O157:H7, regardless of the moisture content in the soil.

Survival and internalization characteristics of Escherichia coli O157:H7 in iceberg, romaine, and leaf lettuce after inoculation of leaf surfaces and soil were determined. A five-strain mixture of E. coli O157:H7 in water and cow manure extract was used as an inoculum for abaxial and adaxial sides of leaves at populations of 6 to 7 log and 4 log CFU per plant. The five strains were individually inoculated into soil at populations of 3 and 6 log CFU/g. Soil, leaves, and roots were analyzed for the presence and population of E. coli O157:H7. Ten (4.7%) of 212 samples of leaves inoculated on the adaxial side were positive for E. coli O157:H7, whereas 38 (17.9%) of 212 samples inoculated on the abaxial side were positive. E. coli O157:H7 survived for at least 25 days on leaf surfaces, with survival greater on the abaxial side of the leaves than on the adaxial side. All 212 rhizosphere samples and 424 surface-sanitized leaf and root samples from plants with inoculated leaves were negative for E. coli O157:H7, regardless of plant age at the time of inoculation or the location on the leaf receiving the inoculum. The pathogen survived in soil for at least 60 days. Five hundred ninety-eight (99.7%) of 600 surface-sanitized leaf and root samples from plants grown in inoculated soil were negative for E. coli O157:H7. Internalization of E. coli O157:H7 in lettuce leaves and roots did not occur, regardless of the type of lettuce, age of plants, or strain of E. coli O157:H7.

The field-core (cut and core) harvesting technique used for iceberg lettuce was evaluated as a potential means of cross-contamination with Escherichia coli O157:H7. Chlorinated water treatment was evaluated for its efficacy in removing or inactivating the pathogen on the blade portion of the field coring device and on cored lettuce. Field coring devices inoculated by immersing blades in soil containing E. coli O157:H7 at 3.74 or 6.57 log CFU/g contained 3.13 and 4.97 log CFU per blade, respectively. Treatment of inoculated field coring device blades by immersing in chlorinated water (200 microg/ml total chlorine) for 10 s resulted in a reduction of 1.56 log CFU per blade, which was 1.42 log CFU per blade greater than that achieved using water, but insufficient to eliminate the pathogen on blades. Field coring devices inoculated by contacting soil containing E. coli O157:H7 at 2.72 and 1.67 log CFU/g, then repeatedly used to cut and core 10 lettuce heads, transferred the pathogen to 10 and 5 consecutively processed heads, respectively. Lettuce cores remained positive for the pathogen after spraying with 100 microg/ml free chlorine for 120 s at 2.81 kg/cm2 (40 lb/in2), regardless of the inoculum level. The number of E. coli O157:H7 recovered from inoculated lettuce cores treated for 10 s with chlorine was significantly (P < or = 0.05) different from the number recovered from tissues treated with water. Dipping contaminated field coring devices in chlorinated water may not be effective in killing the pathogen and controlling cross-contamination from head to head. Spraying contaminated lettuce with chlorinated or untreated water reduces but does not eliminate E. coli O157:H7.

Several outbreaks of Salmonella and Escherichia coli O157:H7 infections have been associated with consumption of leafy greens. Questions remain concerning the ability of these pathogens to become internalized within lettuce and spinach tissues. An effective validated surface disinfection method for lettuce is needed before factors affecting internalization of pathogens can be studied. The objective of this study was to develop a surface disinfection method for lettuce leaves and roots. Iceberg lettuce (Lactuca sativa L.) leaves cut into pieces (3 by 3 cm) and lettuce roots were inoculated by immersing in suspensions of five-strain mixtures of green fluorescent protein-labeled E. coli O157:H7, Salmonella, or Listeria monocytogenes at populations of 7 to 8 log CFU/ml for 10 min at 20 +/- 1 degrees C. Inoculated samples were placed in a laminar flow biosafety cabinet for 30 min before treating with disinfectants. Thirteen surface disinfection methods were compared for their efficacy in killing E. coli O157:H7 on lettuce leaf and root surfaces. E. coli O157:H7 initially at 5.8 or 6.8 log CFU/leaf piece or root was not detected by enumeration (< 0.6 log CFU per leaf piece) on samples treated for 20 min with 10,000 microg/ml sodium hypochlorite (NaHCIO) or in solutions containing ethanol and mercuric chloride (HgCl2). With all other methods, E. coli O157:H7 populations ranged from 2.8 to 4.4 CFU per leaf piece or root after treatment. Trends in leaf and root print and enrichment culture results were consistent with enumeration results. Dipping in 80% ethanol for 10 s followed by immersion in 0.1% HgCl2 for 10 min was determined to be the most effective surface disinfection method for inactivating E. coli O157:H7 on lettuce leaves and roots and was also validated for inactivating Salmonella and L. monocytogenes.

In Fall 2006, four separate outbreaks of foodborne illness associated with the consumption of fresh produce occurred in the United States. In follow-up investigations, spinach, lettuce, and tomatoes were identified as the vehicles of illness. Epidemiologic investigations subsequently focused on finding the specific growing regions using traceback records. While the areas most likely involved in the outbreaks have been identified, the specific mode of contamination remains unconfirmed. Suspected risk factors in these cases include: proximity of irrigation wells and surface waterways exposed to faeces from cattle and wildlife; exposure in fields to wild animals and their waste materials; and improperly composted animal manure used as fertilizer. Difficulty in deciphering these and other on-farm routes of contamination is due to the sporadic nature of these events. Hence, evidence to support these contamination modes is based largely on experimental studies in the laboratory and field. Still at issue is the relevance of internalization of pathogens, whether this occurs through the roots and plant vascular tissues of vegetables and fruits or through plant surfaces into cracks and crevices. Potential for these events, conditions under which the events occur, and pathogen survival following these events, are questions that still need to be answered. Answers to these questions will ultimately affect the type of interventions needed for application postharvest. Currently, many chemical and biological interventions can reduce surface pathogens and minimize cross-contamination, however, they are largely ineffective on internalized pathogens. In the event internalization is a significant route of contamination in the field, physical interventions (irradiation and high pressure) may be needed to minimize risk. Ultimately, risk assessment studies will be useful tools in developing risk management strategies for the produce industry.

Studies were conducted to determine the fate of Escherichia coli O157:H7 in dairy manure-based compost in a field setting. Two trials were performed involving duplicate compost heaps constructed at an outdoor fenced site. The compost heaps were composed of dairy manure, old hay, feed waste, a mixture of sawdust and calf feces, and fresh hay. Samples of the composting mixture were inoculated with stx-negative E. coli O157:H7 B6914 at initial concentrations of 10(7) and 10(5) CFU/g for trial 1 and trial 2, respectively. Individual sample bags were placed on the surface and at three locations (top, center, and bottom) within each heap. Although the compost heaps achieved temperatures of 50 degrees C or above at all internal locations for at least 7 days, temperature stratification was observed. In trial 1, E. coli O157:H7 was detected by enrichment through 14 days within the heaps. When inoculated with 10(5) CFU/g in trial 2, E. coli O157:H7 was detected only through days 2, 2, and 5 at the top, center, and bottom locations, respectively. For both trials, the pathogen survived at the heap's surface for up to 4 months. The indicator commensal E. coli and coliforms were inactivated at a rate similar to that for E. coli O157:H7. Results indicate that composting, with periodic heap turning, can be a practical approach to inactivating E. coli O157:H7 in cattle wastes on the farm. Our data also suggest when compost heaps are not turned, E. coli O157:H7 may survive for months at the heap surface.

Contaminated food continues to be the principal vehicle for transmission of Escherichia coli O157:H7 and other Shiga toxin-producing E. coli (STEC) to humans. A large number of foods, including those associated with outbreaks (alfalfa sprouts, fresh produce, beef, and unpasteurized juices), have been the focus of intensive research studies in the past few years (2003 to 2006) to assess the prevalence and identify effective intervention and inactivation treatments for these pathogens. Recent analyses of retail foods in the United States revealed E. coli O157:H7 was present in 1.5% of alfalfa sprouts and 0.17% of ground beef but not in some other foods examined. Differences in virulence patterns (presence of both stx1 and stx2 genes versus one stx gene) have been observed among isolates from beef samples obtained at the processing plant compared with retail outlets. Research has continued to examine survival and growth of STEC in foods, with several models being developed to predict the behavior of the pathogen under a wide range of environmental conditions. In an effort to develop effective strategies to minimize contamination, several influential factors are being addressed, including elucidating the underlying mechanism for attachment and penetration of STEC into foods and determining the role of handling practices and processing operations on cross-contamination between foods. Reports of some alternative nonthermal processing treatments (high pressure, pulsed-electric field, ionizing radiation, UV radiation, and ultrasound) indicate potential for inactivating STEC with minimal alteration to sensory and nutrient characteristics. Antimicrobials (e.g., organic acids, oxidizing agents, cetylpyridinium chloride, bacteriocins, acidified sodium chlorite, natural extracts) have varying degrees of efficacy as preservatives or sanitizing agents on produce, meat, and unpasteurized juices. Multiple-hurdle or sequential intervention treatments have the greatest potential to minimize transmission of STEC in foods.

Protozoan parasites can survive under ambient and refrigerated storage conditions when associated with a range of substrates. Consequently, various treatments have been used to inactivate protozoan parasites (Giardia, Cryptosporidium, and Cyclospora) in food, water, and environmental systems. Physical treatments that affect survival or removal of protozoan parasites include freezing, heating, filtration, sedimentation, UV light, irradiation, high pressure, and ultrasound. Ozone is a more effective chemical disinfectant than chlorine or chlorine dioxide for inactivation of protozoan parasites in water systems. However, sequential inactivation treatments can optimize existing treatments through synergistic effects. Careful selection of methods to evaluate inactivation treatments is needed because many studies that have employed vital dye stains and in vitro excystation have produced underestimations of the effectiveness of these treatments.

Avian influenza viruses and antibiotic-resistant pathogens have become topics of current public health interest. This paper will focus on the significance of these pathogens to the meat industry as well as other emerging microbiological food safety topics likely to impact the meat industry. These include surveillance of foodborne pathogens, microbial source tracking, risk assessment, and human populations at increased risk of infection by foodborne microbes. These emerging issues will likely lead to even greater challenges to producing microbiologically safe meat products than the industry has ever experienced. However, accompanying such challenges will be innovative solutions that provide even greater public health protection to meat-containing foods.

Several foodborne pathogens, including Salmonella species and campylobacters, are common contaminants in poultry and livestock. Typically, these pathogens are carried in the animal's intestinal tract asymptomatically; however, they can be shed in feces in large populations and be transmitted by other vectors from feces to animals, produce, or humans. A wide array of interventions has been developed to reduce the carriage of foodborne pathogens in poultry and livestock, including genetic selection of animals resistant to colonization, treatments to prevent vertical transmission of enteric pathogens, sanitation practices to prevent contamination on the farm and during transportation, elimination of pathogens from feed and water, feed and water additives that create an adverse environment for colonization by the pathogen, and biological treatments that directly or indirectly inactivate the pathogen within the host. To successfully reduce the carriage of foodborne pathogens, it is likely that a combination of intervention strategies will be required.