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In 2019, citrus production in Florida declined by more than 70%, mostly because of Huanglongbing (HLB), which is caused by the bacterium 'Candidatus Liberibacter asiaticus' (CLas). Thermotherapy for HLB-affected trees was proposed as a short-term management solution to maintain field productivity. It was hypothesized that thermotherapy could eliminate HLB from affected branches; therefore, the study objectives were to show which time-temperature combinations eliminated CLas from woody tissues. Hardening, rounded Valencia twigs collected from HLB-affected field trees were treated in a steam chamber at different time-temperature combinations (50°C for 60 s; 55°C for 0, 30, 60, 90, and 120 s; 60°C for 30 s; and an untreated control). Three independent repetitions of 13 branches per treatment were grafted onto healthy rootstocks and tested to detect CLas after 6, 9, and 12 months. For the RNA-based CLas viability assay, three branches per treatment were treated and bark samples were peeled for RNA extraction and subsequent gene expression analyses. During the grafting study, at 12 months after grafting, a very low frequency of trees grafted with twigs treated at 55%#176;C for 90 s and 55°C for 120 s had detectable CLas DNA. In the few individuals with CLas, titers were significantly lower (P < 0.0001) and could have been remnants of degrading DNA. Additionally, there was a significant decrease (P > 0.0001) in CLas 16S rRNA expression at 55°C for 90 s, 55°C for 120 s, and 60°C for 30 s (3.4-fold change, 3.4-fold change, and 2.3-fold change, respectively) in samples 5 days after treatment. Heat injury, not total CLas kill, could explain the limited changes in transcriptional activity; however, failed recovery and eventual death of CLas resulted in no CLas detection in most of the grafted trees treated with the highest temperatures or longest durations. [ABSTRACT FROM AUTHOR]

Agricultural water is considered as one of the main contamination source for produce prior to harvest. The purpose of study was to evaluate the fate of Shiga toxin-producing Escherichia coli (STEC), and generic E. coli in Central Florida agricultural surface water at different temperatures and the potential use of EPA Worst Case water as a standardized media. Cocktails of STEC (O145, O104, O111, O103, O157), and generic E. coli K-12 were inoculated into agricultural surface water samples (non-sterile and sterilized) and EPA Worst Case water, and enumerated for up to 168 days. E. coli was held at 15 and 25 ± 1 °C. Tested microorganisms decreased most rapidly in non-sterile surface water. At day 168, E. coli populations decreased to ≤2.5 log CFU/100 ml in non-sterile surface water and were 4.8 ≤ and ≤ 8.5 log CFU/100 ml in sterile surface water and EPA Worst Case water. Populations were significantly (P ≤.05) higher in sterile surface water and EPA Worst Case water at all sampling points starting from Day 28. Rate of declines in non-sterile surface waters were between 32.8 and 50 days at both tested temperatures and microorganisms. Addition of cycloheximide to non-sterile surface waters resulted in no significant effect on behavior of E. coli populations. Monitoring generic E. coli (represented by K-12) population changes is a reasonable indicator of STEC survival in agricultural water. EPA Worst Case water is a suitable standard control for surface water microcosms. Unlabelled Image • Survival of STEC, and generic E. coli was studied in surface waters. • STEC and generic E. coli stayed alive in non-sterile microcosms for 168 days. • STEC and generic E. coli showed similar survival trends at all conditions. • EPA Worst Case water offers fair performance as a standard to compare studies. [ABSTRACT FROM AUTHOR]

Abstract The US Produce Safety Rule allows for use of water that does not meet its microbial standards if corrective measures are employed. This research was initiated to determine the suitability of nonpathogenic Escherichia coli as a surrogate for Salmonella during citrus washing, and to evaluate the removal of E. coli from grapefruit on two pilot packinglines (CREC and IRREC) as corrective measures. Whole grapefruit were inoculated with either E. coli or Salmonella and dried, and exposed to a variety of treatments on a lab-scale brush wash system. Individual processes were evaluated on the pilot packinglines with E. coli only. In all lab-scale brush wash system treatments, bacterial population reductions between E. coli and Salmonella were not significantly different (P ≤ 0.05). On pilot packinglines, E. coli populations were reduced by 3.59 to >5.11 log CFU/grapefruit at the CREC packingline, and by 3.30 to >5.13 log CFU/grapefruit at the IRREC packingline. Treatment of fruit through complete packingline processing at both locations reduced E. coli populations to levels below the detection limit (<1 log CFU/grapefruit). The studies indicate E. coli is an appropriate surrogate for Salmonella under tested conditions, and that standard citrus packingline processes can be used as a corrective measure. Highlights • E. coli was a suitable surrogate for Salmonella on grapefruit packinglines. • Reductions of E. coli and background occurred using common postharvest treatments. • Packingline processes reduced E. coli populations from 2.1 to >5.0 log CFU/fruit. • Background populations were reduced from 0.6 to 4.0 log CFU/grapefruit. [ABSTRACT FROM AUTHOR]

This study quantifies the cross-contamination rates between fresh-cut produce and hands using a nalidixic acid–resistant nonpathogenic Enterobacter aerogenes and cocktails of rifampin-resistant Salmonella or Escherichia coli O157:H7 strains. Volunteers performed the E. aerogenes experiments (n = 20), and one of the authors performed the Salmonella and E. coli O157:H7 experiments multiple times (n =15 and n =10, respectively). Each participant handled 25 g of fresh-cut carrots, celery, or cantaloupe in two different scenarios. In the first scenario, gloved hands were inoculated with 6 log CFU per hand of the bacteria, and in the second scenario, five 25-g pieces of fresh produce were inoculated to a concentration of 6 log CFU/25 g. The glove juice method was used to quantify the bacterial concentration on the gloved hands. About 30% of E. aerogenes on gloved hands was transferred to the carrots and celery, and 18% of E. aerogenes on gloved hands was transferred to the cantaloupe. When carrots or cantaloupe was inoculated with E. aerogenes , 1% was transferred to gloved hands; from inoculated celery, about 0.3% of E. aerogenes was transferred to gloved hands. There was not a significant difference between E. aerogenes and Salmonella cross-contamination rates (P > 0.05). When gloved hands were contaminated with E. coli O157:H7, about 30% was transferred to carrots, about 10% to celery, and about 3% to cantaloupe. When carrots and celery were inoculated with E. coli O157:H7, about 1% was transferred to gloved hands, but from inoculated cantaloupe only about 0.3% was transferred. Direction of transfer (to versus from produce), difference in type of produce, and differences among the bacterial species all had significant effects on the transfer rate. Understanding transfer rates to and from fresh-cut produce will allow for better risk assessment and management of microbial food safety risk related to fresh-cut produce. [ABSTRACT FROM AUTHOR]

All published D- values for Shiga toxin-producing Escherichia coli (STEC), Salmonella , and Listeria monocytogenes in orange juice were obtained using strain cocktails. The objective of this study was to evaluate the heat resistance of individual strain of stationary phase non-adapted and acid adapted STEC, Salmonella spp., and L. monocytogenes in orange juice. Three STEC and Salmonella isolates were grown in TSB, and three L. monocytogenes strains grown in BHI, supplemented with 1% glucose for acid adaption. Sealed microcapillary tubes with inoculated single-strength pasteurized orange juice without pulp were treated at 56, 58, and 60 °C for STEC and L. monocytogenes and at 55, 58, and 60 °C for Salmonella . Thermal tolerance was increased significantly ( P < 0.05) for acid adapted STEC strains, however, no improvement was observed for Salmonella spp., and L. monocytogenes strains at most temperatures tested. Salmonella serotypes are less heat resistant, at all temperatures tested, than L. monocytogenes and STEC. STEC, especially strain O111, are the most heat resistant at 56 and 58 °C; L. monocytogenes strains are the most thermal tolerance at 60 °C. Combining individual results of all pathogens tested, the formula of log D = 8.2167 − 0.1356 T (°C) was used to calculate a general process for orange juice at 71.1 °C. Using this equation, a 5-log reduction of all three pathogens in single strength orange juice requires 11 s at 71.1 °C, with a z -value of 7.1 °C. [ABSTRACT FROM AUTHOR]

The purpose of this study was to evaluate the fate of generic Escherichia coli in fecal pats under different field conditions and to predict Salmonella and Shiga toxin-producing E. coli (STEC) survival dynamics based on developed models. Eight trials were conducted during spring and fall in both North and Central Florida. Fresh cattle feces (1g) was placed on mature green round tomatoes located inside (TIP) and outside (TOP) of the plant canopy. Fecal pats (10 g) were placed under (MUP) and distant (MDP) to tomato plants on plastic mulch. Pathogen populations were predicted based on developed models. Declines in generic E. coli populations over 7 days (Log CFU/g) in fecal pats were between 0.9 and 2.7 on TIP, 1.2 and 3.0 on TOP, 0.2 and 1.2 on MUP, and 0.4 and 1.5 on MDP in the Central Florida fall and spring trials, respectively. E. coli populations remained stable at ≤4.2 and ≤ 6.5 Log CFU/g during all North Florida trials. The concentration changes in predicted Salmonella and STEC population were less than 2.1 Log CFU/g in fecal pats for all conditions. Developed models predicted similar pathogen survival trends to generic E. coli with no dramatical impact under the field conditions. • E. coli population in cow pats on tomato and mulch stayed high in the field. • Location of fruit in the plant canopy did not affect E. coli survival in fecal pats. • Fecal pats on mulch in shade had similar E. coli population to ones under sunlight. • Models predict limited changes in pathogen populations in the field conditions. [ABSTRACT FROM AUTHOR]

This study evaluated the efficacy of aqueous (aQUAT) and isopropyl alcohol–based quaternary ammonium (ipQUAT) sanitizers for reducing Salmonella spp., Escherichia coli O157:H7, or Listeria monocytogenes populations on peanut and pistachio shell pieces. Inoculated nutshells were mixed with QUAT sanitizers, water, or 70% ethanol and enumerated immediately or after incubation at 30 °C for 48 h. None of the treatments had any immediate effect on Salmonella or E. coli O157:H7 populations on the peanut or pistachio shells. L. monocytogenes populations declined immediately on the peanut and pistachio shells treated with aQUAT or ipQUAT. After incubation, Salmonella and E. coli O157:H7 populations increased significantly on the water- or aQUAT-treated peanut and pistachio shells. L . monocytogenes populations also increased significantly on the water- or aQUAT-treated peanut shells, but levels did not change on the water-treated pistachio shells and levels were just above the limit of detection on the aQUAT-treated pistachio shells. After treatment with ipQUAT and 48-h incubation, Salmonella and E. coli O157:H7 populations decreased to or below the limit of detection on both shell types; L. monocytogenes populations remained at or below the limit of detection on both shell types. [ABSTRACT FROM AUTHOR]

Hand washing is recognized as a crucial step in preventing foodborne disease transmission by mitigating cross-contamination among hands, surfaces, and foods. This research was undertaken to establish the importance of several keys factors (soap, soil, time, and drying method) in reducing microorganisms during hand washing. A nonpathogenic nalidixic acid-resistant Enterobacter aerogenes surrogate for Salmonella was used to assess the efficacy of using soap or no soap for 5 or 20 s on hands with or without ground beef debris and drying with paper towel or air. Each experiment consisted of 20 replicates, each from a different individual with ~6 log CFU/ml E. aerogenes on their hands. A reduction of 1.0 ± 0.4 and 1.7 ± 0.8 log CFU of E. aerogenes was observed for a 5-s wash with no soap and a 20-s wash with soap, respectively. When there was no debris on the hands, there was no significant difference between washing with and without soap for 20 s (P > 0.05). Likewise, there was no significant difference in the reductions achieved when washing without soap, whether or not debris was on the hands (P > 0.05). A significantly greater reduction (P < 0.05) in E. aerogenes (0.5 log CFU greater reduction) was observed with soap when there was ground beef debris on the hands. The greatest difference (1.1 log CFU greater average reduction) in effectiveness occurred when ground beef debris was on the hands and a 20-s wash with water was compared with a 20-s wash with soap. Significantly greater (P < 0.05) reductions were observed with paper towel drying compared with air (0.5 log CFU greater reductions). Used paper towels may contain high bacterial levels (>4.0 log CFU per towel) when hands are highly contaminated. Our results support future quantitative microbial risk assessments needed to effectively manage risks of foodborne illness in which food workers' hands are a primary cause. [ABSTRACT FROM AUTHOR]

Salmonella accounts for approximately 50% of produce-associated outbreaks in the United States, several of which have been traced back to contamination in the produce production environment. To quantify Salmonella diversity and aid in identification of Salmonella contamination sources, we characterized Salmonella isolates from two geographically diverse produce-growing regions in the United States. Initially, we characterized the Salmonella serotype and subtype diversity associated with 1,677 samples collected from 33 produce farms in New York State (NYS). Among these 1,677 samples, 74 were Salmonella positive, yielding 80 unique isolates (from 147 total isolates), which represented 14 serovars and 23 different pulsed-field gel electrophoresis (PFGE) types. To explore regional Salmonella diversity associated with production environments, we collected a smaller set of samples (n = 65) from South Florida (SFL) production environments and compared the Salmonella diversity associated with these samples with the diversity found among NYS production environments. Among these 65 samples, 23 were Salmonella positive, yielding 32 unique isolates (from 81 total isolates), which represented 11 serovars and 17 different PFGE types. The most common serovars isolated in NYS were Salmonella enterica serovars Newport, Cerro, and Thompson, while common serovars isolated in SFL were Salmonella serovars Saphra and Newport and S. enterica subsp. diarizonae serovar 50:r:z. High PFGE type diversity (Simpson's diversity index, 0.90 ± 0.02) was observed among Salmonella isolates across both regions; only three PFGE types were shared between the two regions. The probability of three or fewer shared PFGE types was <0.000001; therefore, Salmonella isolates were considerably different between the two sampled regions. These findings suggest the potential for PFGE-based source tracking of Salmonella in production environments. [ABSTRACT FROM AUTHOR]

A recent outbreak linked to whole cantaloupes underscores the importance of understanding growth kinetics of Listeria monocytogenes in cut melons at different temperatures. Whole cantaloupe, watermelon, and honeydew purchased from a local supermarket were cut into 10 ± 1 g cubes. A four-strain cocktail of L. monocytogenes from food related outbreaks was used to inoculate fruit, resulting in ∼103 CFU/10 g. Samples were stored at 4, 10, 15, 20, or 25 °C and L. monocytogenes were enumerated at appropriate time intervals. The square root model was used to describe L. monocytogenes growth rate as a function of temperature. The model was compared to prior models for Salmonella and Escherichia coli O157:H7 growth on cut melon, as well as models for L. monocytogenes on cantaloupe and L. monocytogenes ComBase models. The current model predicts faster growth of L. monocytogenes vs. Salmonella and E. coli O157:H7 at temperatures below 20 °C, and agrees with estimates from ComBase Predictor, and a corrected published model for L. monocytogenes on cut cantaloupe. The model predicts ∼4 log CFU increase following 15 days at 5 °C, and ∼1 log CFU increase following 6 days at 4 °C. The model can also be used in subsequent quantitative microbial risk assessments. [ABSTRACT FROM AUTHOR]

Abstract: In natural-light labeling of fruits and vegetables, the desired information is etched onto the produce surface using a low-energy carbon dioxide laser beam (10,600 nm). Etched characters are formed by surface depressions in the epidermis that may facilitate entrance of decay and pathogenic organisms. The objective of this study was to determine the effects of natural-light labeling and different postharvest treatments on Salmonella populations' ability to survive/grow and penetrate into citrus fruit. A five-strain cocktail of Salmonella was spot inoculated onto Valencia orange in different application sequences with wax and natural-light etching. Samples were stored at 10, 26 °C, or combinations of both, for up to 42 days. Etched peels and corresponding juices were extracted from whole oranges following storage and enumerated for Salmonella. No set of conditions involving natural-light labeling promoted the growth of Salmonella on the fruit surface or resulted in the detection of Salmonella from the juice of sound fruit. Survival of Salmonella populations on the peel surface did not differ between any of the treatment and control samples. In all cases, Salmonella declined between 1.5 and 3.0 log CFU/orange after 30 days, with faster decline noted at 10 °C. Based on the data obtained from all treatments and under conditions extremely unfavorable and unrealistic in terms of fruit storage, natural-light labeling citrus fruit peels and subsequent waxing in any order did not allow for the growth or influence the natural decline of Salmonella populations on citrus fruit surfaces, or movement into juices, as compared to controls. [Copyright &y& Elsevier]

Outbreaks of Salmonella enterica have increasingly been associated with tomatoes and traced back to production areas, but the spread of Salmonella from a point source onto plants has not been described. Splash dispersal by rain could be one means of dissemination. Green fluorescent protein-labeled, kanamycin-resistant Salmonella enterica sv. Typhimurium dispensed on the surface of plastic mulch, organic mulch, or soil at 108 CFU/cm² was used as the point source in the center of a rain simulator. Tomato plants in soil with and without plastic or organic mulch were placed around the point source, and rain intensities of 60 and 110 mm/h were applied for 5, 10, 20, and 30 min. Dispersal of Salmonella followed a negative exponential model with a half distance of 3 cm at 110 mm/h. Dispersed Salmonella survived for 3 days on tomato leaflets, with a total decline of 5 log and an initial decimal reduction time of 10 h. Recovery of dispersed Salmonella from plants at the maximum observed distance ranged from 3 CFU/g of leaflet after a rain episode of 110 mm/h for 10 min on soil to 117 CFU/g of leaflet on plastic mulch. Dispersal of Salmonella on plants with and without mulch was significantly enhanced by increasing rain duration from 0 to 10 min, but dispersal was reduced when rainfall duration increased from 10 to 30 min. Salmonella may be dispersed by rain to contaminate tomato plants in the field, especially during rain events of 10 min and when plastic mulch is used. [ABSTRACT FROM AUTHOR]

Abstract: Salmonellosis outbreaks from consumption of raw almonds in 2001 and 2004 led to regulations that require mandatory treatment of almonds sold in North America to give a minimum 4-log reduction of Salmonella. This study aims to: 1) assess the risk of salmonellosis associated with almond consumption in North America, with current treatments in effect; 2) determine the resilience of the current production system to increases in prevalence or concentration of Salmonella on almonds; 3) assess the impact of treating less than 100% of the crop; and 4) investigate conditions that could explain the number of cases associated with the 2001 outbreak. Risk was assessed using a Monte Carlo simulation, based on an established dose–response relationship. Data for almond amounts sold, Salmonella prevalence and concentration on almonds, storage time and temperature at different handling steps, population reductions during storage at various temperatures and with different treatments, and consumer handling were based on data from published sources and almond industry or academic expert opinion. What-if scenarios were evaluated for Salmonella prevalence varying from 1 to 65%, concentrations of Salmonella varying from 1 to 120MPN/100g, and portions of untreated crop varying from 0 to 10%. The estimated incidence of salmonellosis in North America from almonds as currently treated is on average 0.008 cases per billion servings (with an estimated 6.6billion servings consumed annually). Increases in Salmonella prevalence to 25%, mean concentrations above 25MPN/100g, or leaving 0.05% of the crop untreated all resulted in an arithmetic mean risk greater than 1case/year (with geometric means remaining below 1case/year for all variables). Assuming 4000kg at a prevalence of 65% (observed in recalled lots) and an average concentration of 120MPN/100g in raw almonds (back calculated from levels in recalled almonds) predicted over the 2800 cases estimated for the 2001 outbreak. Applying a 4-log reduction to these almonds reduced the average number of predicted cases to less than a single case. The current regulation is effective in maintaining the risk of salmonellosis from consumption of almonds below an arithmetic mean of 1case/year, although significant increases in either prevalence or concentration, or small increases in proportion of untreated almonds would frequently lead to exceeding this threshold. [Copyright &y& Elsevier]

Abstract: Guaiacol is a profoundly negative taint/off-flavour produced by the increasingly common microbial contamination of fruit juices with Alicyclobacillus spp. The objectives of this study included: determining sensory thresholds for guaiacol in orange juice (OJ), developing an analytical method whose detection limits were equivalent to sensory thresholds and determining levels of Alicyclobacillus spp. that would produce detectable levels of guaiacol. A 12 member trained panel was used to establish guaiacol detection and recognition thresholds. Guaiacol ortho and retro nasal detection thresholds in OJ were 0.70 and 0.53μg/l respectively. Odour recognition threshold was 2.0μg/l. A SPME GC–MS Selected Ion Monitoring (SIM) procedure was optimised to achieve analytical detection limits of 0.5μg/l. Optimum guaiacol detection limit was achieved using only responses from m/z 109 and 124. Ion ratios (m/z 109/124) and linear retention index value matching were used to confirm the identification of guaiacol. Quantification was achieved using peak areas from standard guaiacol additions in orange juice between 0.5 and 100μg/l. Alicyclobacillus growth of 2.2 log CFU/ml in OJ produced just detectable levels (0.7μg/l) of guaiacol. [Copyright &y& Elsevier]

This project was undertaken to relate what is known about the behavior of Escherichia coli O157:H7 under laboratory conditions and integrate this information to what is known regarding the 2006 E. coli O157:H7 spinach outbreak in the context of a quantitative microbial risk assessment. The risk model explicitly assumes that all contamination arises from exposure in the field. Extracted data, models, and user inputs were entered into an Excel spreadsheet, and the modeling software @RISK was used to perform Monte Carlo simulations. The model predicts that cut leafy greens that are temperature abused will support the growth of E. coli O157:H7, and populations of the organism may increase by as much a 1 log CFU/day under optimal temperature conditions. When the risk model used a starting level of -1 log CFU/g, with 0.1% of incoming servings contaminated, the predicted numbers of cells per serving were within the range of best available estimates of pathogen levels during the outbreak. The model predicts that levels in the field of - 1 log CFU/g and 0.1% prevalence could have resulted in an outbreak approximately the size of the 2006 E. coli O157:H7 outbreak. This quantitative microbial risk assessment model represents a preliminary framework that identifies available data and provides initial risk estimates for pathogenic E. coli in leafy greens. Data gaps include retail storage times, correlations between storage time and temperature, determining the importance of E. coli O157:H7 in leafy greens lag time models, and validation of the importance of cross-contamination during the washing process. [ABSTRACT FROM AUTHOR]

A metabolomic-based method for rapid detection of Escherichia coli O157:H7, Salmonella Hartford, Salmonella Typhimurium, and Salmonella Muenchen in nonselective media was developed. All pathogenic bacteria were grown in tryptic soy broth (TSB) at 37 °C followed by metabolite quantification at 2-h intervals for 24 h. Results were compared with the metabolite profiles similarly obtained with E. coli K12, Pseudomonas aeruginosa, Staphylococcus aureus, Saccharomyces cereviseae, and Aspergillus oryzae grown individually or as a cocktail under the same conditions. Principal component analysis (PCAS) discriminated pathogenic microorganisms grown in TSB. Metabolites responsible of PCAS classification were dextrose, cadaverine, the aminoacids L-histidine, glycine, and L-tyrosine, as well as the volatiles 1-octanol, 1-propanol, 1butanol, 2-ethyl-1-hexanol, and 2,5-dimethyl-pyrazine. Partial least square (PLS) models based on the overall metabolite profile of each bacteria were able to detect the presence of Escherichia coli O157:H7 and Salmonella spp. at levels of approximately 7 ± 2 CFU/25 g of ground beef and chicken within 18 h. Rapid detection of foodborne pathogens is critical to reduce food-related disease outbreaks and product recalls. This research proposes an alternative approach to rapidly detect 2 major pathogens based on detecting changes in the metabolite profile during incubation in nonselective culture media. Although not directly transferable to an industrial environment, this proof of concept research provides evidence of potential application. [ABSTRACT FROM AUTHOR]

Abstract: The presence of Alicyclobacillus in fruit juices and concentrates poses a serious problem for the juice industry. This study was undertaken to determine the (i) prevalence, concentration, and species of Alicyclobacillus in tropical and subtropical concentrates; (ii) efficacy of aqueous chlorine dioxide in reducing Alicyclobacillus spp. spores on tropical and subtropical fruit surfaces; and (iii) fate of and off-flavor production by Alicyclobacillus acidoterrestris in mango and pineapple juices. One hundred and eighty tropical and subtropical juice concentrates were screened for the presence and concentration of Alicyclobacillus spp. If found, the species of Alicyclobacillus was determined by 16S rDNA sequencing and analysis with NCI BLAST. Of these samples, 6.1% were positive for Alicyclobacillus, and nine A. acidoterrestris strains and two Alicyclobacillus acidocaldarius strains were identified. A five-strain cocktail of Alicyclobacillus spp. was inoculated onto the surface of fruits (grapefruit, guava, limes, mangoes, oranges and pineapple), which were then washed with 0, 50, or 100 ppm aqueous chlorine dioxide. Significant reductions due to chlorine dioxide were only seen on citrus fruits. A five-strain cocktail of A. acidoterrestris was inoculated into mango and pineapple juices. Microbial populations were enumerated over a 16-day period. Aroma compounds in the juice were analyzed by GC–olfactometry (GC–O) and confirmed using GC–MS. GC–O of mango juice identified previously reported medicinal/antiseptic compounds. GC–O of pineapple juice revealed an unexpected “cheese” off-aroma associated with 2-methylbutyric acid and 3-methylbutyric acid. [Copyright &y& Elsevier]

The survival-growth capacity of Salmonella populations on tomato epidermis labeled by a natural-light labeling system was investigated after persistent fears of such marks serving as possible entryways for the pathogenic organisms, alone and in the presence of Pectobacterium carotovorum subsp. carotovorum, a soft-rot organism. Different treatments involving natural-light labeling, fruit waxing, and a five-strain cocktail of Salmonella were applied to mature green tomato surfaces in different sequences prior to storage at 4, 12, or 25°C. Fruit was sampled every 3 days, and Salmonella was enumerated from all treatments and unlabeled fruit, which served as controls. There were no significant differences between treatments or between treatments and controls throughout. The results indicate that the cuticle and epidermal interruptions caused by natural-light labeling do not facilitate the penetration and colonization of the tomato pericarp. In a separate set of experiments, the capacity of Salmonella to penetrate tomato in the presence of a potential synergism with P. carotovorum subsp. carotovorum was investigated. The addition of P. carotovorum at higher, lower, or equal population densities to Salmonella did not significantly alter the behavior of Salmonella on tomatoes stored at 25°C, regardless of natural-light labeling. The inability of P. carotovorum and Salmonella to colonize natural-light-etched surfaces of tomato fruit indicates that the use of this technology does not adversely compromise the surface of tomatoes. [ABSTRACT FROM AUTHOR]

The adoption of mechanical harvesting for processing oranges is a major objective of the Florida citrus industry. A number of issues have slowed the adoption of this new technology, including the observation that the amount of leaves, stems, and dead branches (collectively termed "debris") is greater in mechanically harvested than in hand-harvested loads of fruit. This debris increases transportation and processing costs. The objective of this research was to determine the amount and types of debris in mechanically harvested loads of sweet oranges compared with hand-harvested controls. Mechanical harvesting was found to increase the amount of debris per load of fruit by as much as 250% compared with hand-harvested fruit. This translates into ≈ 108 kg of debris compared with 71 kg (fresh weight) per 27 t load for mechanically harvested and hand-harvested fruit, respectively. Across harvesting method, leaves were the largest component of debris, accounting for ≈60% of total debris, small stems (less than 5 mm diameter) accounted for ≈35%, and the remaining 5% was large stems (greater than 5 mm diameter). In addition, the amount of sand on the surface of mechanically harvested fruit that was picked up from the orchard floor was found to be up to 10 times greater compared with hand-harvested controls. Engineers developing debris elimination systems for mechanical harvesting systems can use the data from this study to determine the performance requirements of their systems. The data are also useful for economic analyses of the costs of mechanical harvesting. [ABSTRACT FROM AUTHOR]

Abstract: Escherichia coli O157:H7 and Salmonella infections have been associated with consumption of a number of fruits and vegetables. Although the fate of E. coli O157:H7 and Salmonella on many of these products is well studied, little is known about their behavior on cut mango and papaya. Mangoes and papayas have been associated with four and two documented outbreaks of salmonellosis, respectively. The objective of this study was to evaluate the fate of E. coli O157:H7 and Salmonella on fresh (4°C, 12°C and 23°C) and frozen (−20°C) cut mangoes and papayas. Cut mangoes and papayas were spot inoculated with either a four-strain or five-strain cocktail of E. coli O157:H7 or Salmonella, respectively. Inoculated samples were air dried, placed in containers and stored at 4±2, 12±2, 23±2 and −20±2°C. Samples were enumerated following stomaching on nonselective and selective media at days 0, 1, 3, 5, 7, 10, 14, 21 and 28 (4±2 and 12±2°C); 0, 1, 3, 5 and 7 (23±2°C); and 0, 7, 14, 21, 28, 60, 90, 120, 150 and 180 (−20±2°C). Population levels (log CFU/g) of fruit were calculated. E. coli O157:H7 and Salmonella have the potential to grow on temperature-abused fresh cut mangoes and papayas held at 23°C. At 12°C, Salmonella populations can grow on cut mangoes and papayas, however E. coli O157:H7 populations only grew on papayas. E. coli O157:H7 and Salmonella survived for 28days, at 4°C, on refrigerated mangoes and papayas. E. coli O157:H7 and Salmonella can survive on frozen cut mangoes and papayas for at least 180days. Fresh and frozen cut mangoes and papayas are potential vectors for E. coli O157:H7 and Salmonella transmission. [Copyright &y& Elsevier]

Pineapples have been associated with outbreaks of Escherichia coli and Salmonella. Little is known about the behavior of E. coli O157:H7 and Salmonella on cut pineapple. The objectives of this study were to evaluate the fate of E. coli O157:H7 and Salmonella on fresh and frozen cut pineapples and the influence of acid adaptation on Salmonella survival on fresh cut pineapple at ambient storage (23°C). Cut pineapples were spot inoculated with a four-strain cocktail of E. coli O157:H7 or a five-strain cocktail of Salmonella. Inoculated samples were air dried, placed in containers, and stored at 4, 12, 23, and -20°C. Samples were stomached and then enumerated on nonselective and selective media at days 0, 1, 3, 5, 7, 10, 14, 21, and 28 (4 and 12°C); 0, 1, 3, 5, and 7 (23°C); and 0, 7, 14, 21, 28, 60, 90, 120, 150, and 180 (-20°C). E. coli O157:H7 and Salmonella inoculated onto pineapple did not grow but survived for the shelf life at each temperature, with improved survival at lower temperatures. E. coli O157:H7 and Salmonella can survive on frozen cut pineapples for at least 180 days. Acid adaptation of Salmonella did not allow growth and did not increase survival. Fresh and frozen cut pineapples are potential vectors for E. coli O157:H7 and Salmonella transmission and preventive procedures should be implemented during production and postharvest processing. [ABSTRACT FROM AUTHOR]

The effects due to Candidatus Liberibacter infection, commonly called citrus greening or Huanglongbing (HLB), on volatile and nonvolatile components of orange juices, OJ, were examined using GC-MS and high-performance liquid chromatography (HPLC). HLB symptomatic, asymptomatic, and control “Hamlin” and “Valencia” oranges were harvested from December to May during the 2007 to 2008 harvest season. Brix/acid levels in control and asymptomatic juices were similar but symptomatic juices were as much as 62% lower than control juices. No bitter flavanone neohesperidosides were detected and polymethoxyflavone concentrations were well below bitter taste thresholds. Limonin concentrations were significantly higher (91% to 425%) in symptomatic juice compared to control but still below juice bitterness taste thresholds. Juice terpenes, such as γ-terpinene and α-terpinolene, were as much as 1320% and 62% higher in symptomatic juice than control. Average ethyl butanoate concentrations were 45% lower and average linalool was 356% higher in symptomatic Valencia OJ compared to control. Symptomatic Valencia OJ had on average only 40% the total esters, 48% the total aldehydes, and 33% as much total sesquiterpenes as control juice. Total volatiles between control and symptomatic juices were similar due to elevated levels of alcohols and terpenes in symptomatic juice. There were no consistent differences between asymptomatic and control juices. The chemical composition of juice from HLB/greening symptomatic fruit appears to mimic that of juice from less mature fruit. The reported off-flavor associated with symptomatic juices probably stem from lower concentrations of sugars, higher concentrations of acid as all known citrus bitter compounds were either below taste thresholds or absent. [ABSTRACT FROM AUTHOR]

Large volumes of fine particulate matter or “dust” (soil, hulls, and shells) generated when hulls and shells are removed from almond kernels complicate cleaning and sanitation procedures in the huller-sheller (HS) environment. This study evaluated the efficacy of 3 aqueous quaternary ammonium sanitizers (AQuats) and an isopropyl alcohol-based quaternary ammonium sanitizer (IPAQuat) for reducing Salmonella in dust collected from 2 HS facilities. Dust (1 g) was thoroughly mixed with 1 to 2 mL of inoculum (1 to 5 log CFU/g) before adding 1 to 7 mL of water, an AQuat (200 or 1000 ppm), or IPAQuat (200 ppm, 58.6% isopropyl alcohol) and incubated at 15 and 30 °C for up to 21 d. At either 15 or 30 °C increases in Salmonella populations in the dust were not significantly different following addition of either water or AQuats. No significant differences were observed upon water or AQuat addition, either among the 3 AQuats tested, the concentration or volume of AQuat, or the initial level of Salmonella. When IPAQuat was added to dust inoculated at 1 to 7 log CFU/g, Salmonella levels were reduced to less than 1.3 log CFU/g after treatment and after incubation at 30 °C for 48 h. IPAQuat was an effective sanitizer compared to the AQuats, even in the presence of high levels of organic material. Recent large-scale outbreaks of salmonellosis with low-moisture foods have increased concerns regarding their safety. Little research or guidance is available on appropriate cleaning and sanitation programs for these food types. This research is focused on an evaluation of sanitation options for low-moisture foods, in particular almonds. The information should be applicable and useful to the nut industry and to other low-moisture foods. Practical Application: Recent large-scale outbreaks of salmonellosis with low-moisture foods have increased concerns regarding their safety. Little research or guidance is available on appropriate cleaning and sanitation programs for these food types. This research is focused on an evaluation of sanitation options for low-moisture foods, in particular almonds. The information should be applicable and useful to the nut industry and to other low-moisture foods. [ABSTRACT FROM AUTHOR]

Recent Salmonella outbreaks associated with consumption of fresh produce have increased public concern for the safety of raw food products, perhaps signaling a paradigm shift in approaches to food safety. Limitations to our capacity to ensure that raw foods are safe for the consumer include the availability of sufficiently rapid and reliable technology for prevention, intervention, and risk assessment. Other food products, such as shellfish, with greater historical precedent for real or perceived public health risk may offer perspective and insight into strategies for meeting these challenges. This review documents current practices for pathogen prevention and detection in raw oysters and presents technological advances and impediments that determine the application of these methods. [Copyright &y& Elsevier]

The ability of Salmonella,to migrate from an external aqueous environment through the almond hull and shell, and to colonize the kernel, was evaluated in two ways. First, the outer surface of shell halves from five varieties of almonds that differed in shell hardness were placed in contact with a suspension of Salmonella enterica serovar Enteritidis phage type 30 for 24 h at 24°C. Salmonella Enteritidis was isolated from the inside of these almond shells in 46 and 100% of the samples, by direct swabbing of the inner surface of the shell and by enrichment from the swab, respectively. These findings suggested that hardness of the shell is not a significant factor in the migration of the pathogen through that tissue. In addition, both motile and nonmotile strains of S. enterica serovar Typhimurium migrated through the almond shells to the same extent under the conditions of this assay, indicating that bacterial migration through the wet shell may be a passive process. Second, whole almonds (intact hull, shell, and kernel) were soaked for 24 to 72 h at 24°C in a suspension of Salmonella Enteritidis phage type 30 labeled with the green fluorescent protein. Green fluorescent protein-labeled Salmonella cells were observed on the outer and inner surfaces of both the almond hull and shell, and on the kernel, by confocal laser scanning microscopy. Our data provide direct evidence that wet conditions allow for Salmonella migration through the hull and shell and onto the almond kernel, thus providing a means by which almond kernels may become contaminated in the field. [ABSTRACT FROM AUTHOR]

Studies were conducted to isolate bacteria inhibitory to Salmonella enterica serovar Typhimurium definitive type (DT) 104 in vitro from cattle not carrying Salmonella and to determine the inhibitory activity of the isolated bacteria through competitive growth in cattle feces artificially contaminated with Salmonella Typhimurium DT104 and S. enterica serovar Newport. Fecal samples (108) were obtained from dairy and beef cows. S. enterica serovars were isolated from 9.25% of the samples and included Salmonella Newport (4), Salmonella Bareilly (1), Salmonella Mbandaka (1), Salmonella Montevideo (1), Salmonella Meleagridis (1), and monophasic Salmonella (2). All four Salmonella Newport isolates were resistant to at least nine antibiotics. Of 1,097 bacterial isolates from cattle feces screened, 30 were inhibitory to Salmonella Typhimurium DT104 in vitro. The inhibitory isolates included 22 Escherichia coli, 6 Bacillus circulans, 1 Serratiafonticola, and 1 Enterobacter cloacae. Typing by pulsed-field gel electrophoresis showed 17 distinguishable profiles among the 22 E. coli. Competitive inhibition isolates did not significantly reduce Salmonella Typhimurium DT104 during 21 days of storage at 37°C in cattle feces. B. circulans (105 CFU/g of inoculum) significantly reduced Salmonella Newport on days 3 and 5 and on day 21 with 108 CFU/g of inoculum at 37°C. At 21°C, significant reductions of Salmonella Typhimurium DT104 occurred with 108 CFU of gram-negative competitive inhibition bacteria per g and 105 CFU of B. circulans per g on day 5 only. No significant reductions were observed with Salmonella Newport at 21°C. The 25 competitive inhibition bacteria identified in this study offer a first step in identifying competitive inhibition bacteria that may reduce the level of intestinal carriage and fecal shedding of Salmonella Typhimurium DT104 and Salmonella Newport in cattle. [ABSTRACT FROM AUTHOR]

In 2001, Salmonella Enteritidis phage type (PT) 30 was isolated from drag swabs of 17 61-ha almond orchards on three farms linked to an outbreak of salmonellosis associated with consumption of raw almonds. The objective of this study was to evaluate the long-term persistence of Salmonella Enteritidis PT 30 in one of the almond orchards associated with the outbreak. Swabs (gauze saturated with full-strength sterile evaporated skim milk and attached to string) were pulled along the orchard floor in a standardized manner for 55 m. At each sample time, two pooled samples (four swabs each) were collected from each orchard quadrant. Swabs were enriched for Salmonella using a delayed secondary enrichment procedure developed for isolation of Salmonella from poultry houses. Suspect Salmonella isolates were selected, confirmed, serotyped, and phage typed, and pulsed-field gel electrophoresis (PFGE) patterns were determined after cleavage with XbaI and BlnI. Salmonella was recovered infrequently from pooled samples collected from January through July (3 of 56 samples, 5.3%). In general, Salmonella isolation frequency per sample time increased during and immediately after the harvest, when large amounts of dust were generated in or near the orchard: August, 4 (20%) of 20 samples; September, 13 (20%) of 64 samples; October, 27 (42%) of 64 samples; November, 4 (25%) of 16 samples; and December, 2 (25%) of 8 samples. All 53 Salmonella isolates during the 5 years were identified as Salmonella Enteritidis PT 30, and two PFGE patterns that differed by the presence of an approximately 40-kb fragment were identified. These data demonstrate the potential for long-term environmental persistence of Salmonella in almond orchards. [ABSTRACT FROM AUTHOR]

Data on the prevalence and populations of pathogens in individual foods are critical to the development of product-specific quantitative microbial risk assessments. An outbreak of salmonellosis associated with the consumption of raw almonds in 2000 to 2001 provided an opportunity to evaluate the levels of Salmonella in the recalled product. Duplicate 100-g samples from each of fifty 22.7-kg boxes of recalled almonds were enriched by one of two methods. Salmonella was isolated by at least one method from 42 boxes (84% positive). The levels of Salmonella determined by a three-tube most-probable-number (MPN) method were 8.5 ± 1.3 MPN/100 g. In a subsequent study, raw almonds that arrived at almond processors were sampled from 2001 through 2005 to determine the overall prevalence and levels of Salmonella and to characterize the Salmonella isolates obtained. Aerobic plate counts, coliform counts, and MPN levels of Escherichia coli were also determined on positive samples. An isolation frequency for Salmonella of 81 (0.87% ± 0.2%) of 9,274 samples tested (100 g) was determined for raw almonds sampled from throughout California over the 5-year period. Salmonella was not isolated upon retesting in 59 of 65 positive samples. When detected, levels were 1.2 to 2.9 MPN/100 g. Of the 81 total isolates, 35 different serotypes of Salmonella were represented. Aerobic plate counts, coliform counts, and E. coli levels did not correlate with the presence of Salmonella. [ABSTRACT FROM AUTHOR]

To evaluate die survival of Salmonella on raw almond surfaces, whole almond kernels were inoculated with Salmonella Enteritidis phage type (PT) 30 collected from a 24-h broth culture or by scraping cells from an agar lawn. Kernels inoculated with lawn-collected cells to 8, 5, 3, and 1 log CFU per almond after a 24-h drying period were stored for 161 days at 23 ± 3°C. Calculated rates of reduction were similar for the four inoculum levels (0.22. 0.28, 0.29, and 0.22 log CFU/month, respectively). Kernels inoculated to 7.1 or 8.0 log CFU per almond after drying were stored for 171 or 550 days, respectively, at selected temperatures, including -20 ± 2°C, 4 ± 2°C, 23 ± 3°C, and 35 ± 2°C. No significant reductions of Salmonella were observed during storage at -20 and 4°C over 550 days. At 35°C, a biphasic survival curve was observed, with calculated reductions of 1.1 log CFU/month from days 0 to 59 and no significant reduction from days 59 to 171. At 23°C. reductions of 0.18 and 0.30 log CFU/month were calculated fur 171 and 550 days of storage, respectively. When combined with data from the study of inoculum levels, an overall average calculated reduction at 23°C was 0.25 ± 0.05 log CFU/month. Significantly greater reductions were observed during the 24-h drying period when broth-collected cells were used as the inoculum, suggesting that cells collected from agar lawns were more resistant to drying. However, after initial drying, the rates of reduction at 23°C did not differ significantly between the inoculum preparation methods. Salmonella Enteritidis PT 30 survives for long periods on almond kernels under a variety of common storage conditions. [ABSTRACT FROM AUTHOR]

Recent outbreaks of salmonellosis associated with raw almonds have raised awareness of this food as a vector for foodborne illness. We performed a quantitative assessment of the risk of contracting salmonellosis from consumption of raw almonds, accounting for factors that become important after almonds reach the processor. We estimated the risk associated with the consumption of raw almonds and the risk reduction associated with almonds treated with a theoretical 5-log reduction process or treated with propylene oxide using a standard commercial process. Probability distributions were chosen to describe the chance of almond contamination and the effects of storage time, storage temperature, and processing from currently available data. A β-Poisson model for the dose-response relationship for Salmonella was obtained from published literature. The simulation estimated a 78% chance of one or more cases of salmonellosis per year from consumption of raw almonds. The application of a commercial propylene oxide treatment reduced this risk to 0.01%. Hypothetical 5-log reduction treatments with different standard deviations (±1, ±0.5, ±0.1, and ±0) reduced the predicted yearly risk of salmonellosis to 0.69, 0.35, 0.30, and 0.21%, respectively. These results suggest that the risk of one or more U.S. cases of salmonellosis per year from consumption of raw almonds can be reduced from 78% to less than 1% by using a process achieving a 5-log reduction in Salmonella with a process standard deviation as large as 1 log unit or by using a commercial propylene oxide treatment. [ABSTRACT FROM AUTHOR]

Even though differences in methodology (e.g., sample volume and detection method) have been shown to affect observed microbial water quality, multiple sampling and laboratory protocols continue to be used for water quality monitoring. Research is needed to determine how these differences impact the comparability of findings to generate best management practices and the ability to perform metaanalyses. This study addresses this knowledge gap by compiling and analyzing a data set representing 2,429,990 unique data points on at least one microbial water quality target (e.g., Salmonella presence and Escherichia coli concentration). Variance partitioning analysis was used to quantify the variance in likelihood of detecting each pathogenic target that was uniquely and jointly attributable to non-methodological versus methodological factors. The strength of the association between microbial water quality and select methodological and non-methodological factors was quant ified using conditional forest and regression analysis. Fecal indicator bacteria concentrations were more strongly associated with non-methodological factors than methodological factors based on conditional forest analysis. Variance partitioning analysis could not disentangle non-methodological and methodological signals for pathogenic Escherichia coli, Salmonella, and Listeria. This suggests our current perceptions of foodborne pathogen ecology in water systems are confounded by methodological difference s between studies. For example, 31% of total variance in likelihood of Salmonella detection was explained by methodological and/or non-methodological factors, 18% was jointly attributable to both methodological and non-methodological factors. Only 13% of total variance was uniquely attributable to non-methodological factors for Salmonella, highlighting the need for standardization of methods for microbiological water quality testing for comparison across studies. [ABSTRACT FROM AUTHOR]

Pond irrigation water comprises a major pathway of pathogenic bacteria to fresh produce. Current regulatory methods have been shown to be ineffective in assessing this risk when variability of bacterial concentrations is large. This paper proposes using mechanistic modeling of bacterial transport as a way to identify improved strategies for mitigating this risk pathway. If the mechanistic model is successfully tested against observed data, global sensitivity analysis (GSA) can identify important mechanisms to inform alternative, preventive bacterial control practices. Model development favored parsimony and prediction of peak bacterial concentration events. Data from two highly variable surface water irrigation ponds showed that the model performance was similar or superior to that of existing pathogen transport models, with a Nash-Sutcliffe efficiency of 0.48 and 0.18 for the two ponds. GSA quantified bacterial sourcing and hydrology as the most important processes driving pond bacterial contamination events. Model analysis has two main implications for improved regulatory methods: that peak concentration events are associated with runoff-producing rainfall events and that intercepting bacterial runoff transport may be the best option to prevent bacterial contamination of surface water irrigation ponds and thus fresh produce. This research suggests the need for temporal management strategies. IMPORTANCE Preventive management of agricultural waters requires understanding of the drivers of bacterial contamination events. We propose mechanistic modeling as a way forward to understand and predict such events and have developed and tested a parsimonious model for rain-driven surface runoff contributing to generic Escherichia coli contamination of irrigation ponds in Central Florida. While the model was able to predict the timing of peak events reasonably well, the highly variable magnitude of the peaks was less well predicted. This indicates the need to collect more data on the fecal contamination inputs of these ponds and the use of mechanistic modeling and global sensitivity analysis to identify the most important data needs. [ABSTRACT FROM AUTHOR]

Salmonella persistence in sterile surface water, nonsterile surface water, or deionized water over 336 days was explored. The suitability of EPA (U.S. Environmental Protection Agency) Worst Case water as a standard medium was also evaluated. Salmonella was enumerated by spread plating or most probable number at set time points after inoculation into water (sterile and nonsterile surface, deionized, and EPA Worst Case waters) and held at 15, 21, or 28°C. Total coliforms from nonsterile waters were enumerated by spread plating. Salmonella decreased most rapidly in nonsterile surface water. Addition of cycloheximide to nonsterile surface waters did not increase Salmonella survival compared with the same nonsterile surface water without cycloheximide except for the water held at 15°C. Total coliform populations decreased to below the limit of detection at 112 days; Salmonella remained detectable throughout the entire 168-day experiment. Salmonella populations persist at least 336 days in all waters tested; coliforms remained detectable to 112 days. Rates of decline (time required for a 1-log reduction) in nonsterile waters were at least five times shorter than those of other water types. EPA Worst Case water is a suitable standard for surface water microcosms. This study adds to the limited knowledge regarding Salmonella survival in surface waters. It demonstrates the potential long-term survival of Salmonella in fresh water and the importance of biotic factors when designing microcosm-based experiments. Survival of Salmonella was studied in surface waters. Salmonella stayed alive in nonsterile microcosms for 168 days. Limited decline was seen in sterile surface, deionized, and EPA Worst Case water. EPA Worst Case water offers potential as a standardized medium. [ABSTRACT FROM AUTHOR]

amination of fresh produce (fresh fruits and vegetables) poses serious public health concerns worldwide. This study was conducted as a comprehensive analysis of biological hazards in the global fresh produce chain. Data about producerelated outbreaks and illness were collected from the annual reports and databases of foodborne outbreak surveillance systems in different regions and countries from 2010 to 2015. The global patterns of and regional differences in documented outbreaks and cases were analyzed, and produce commodities and pathogens of greatest concern were identified. Data on sporadic illnesses were also collected through a comprehensive literature review of case-control studies. We found 988 produce-related outbreaks (with known agents) and 45,723 cases in all regions and countries. The numbers of produce-related outbreaks per million personyears were approximately 0.76, 0.26, 0.25, 0.13, 0.12, and 0.05 in New Zealand, Australia, the United States, the European Union, Canada, and Japan, respectively. The top three food categories and pathogens contributing to produce-related outbreaks were vegetables and nonfruits (i.e., food other than fruits; 27.0%), unspecified vegetables (12.2%), and vegetable row crops (11.7%) and norovirus (42.4%), Salmonella enterica (19.9%), and Staphylococcus aureus (7.9%), respectively. Produce consumption was identified as a protective factor, a risk factor, and either a protective or risk factor for sporadic illnesses in 11, 5, and 5 studies, respectively, among 21 case-control studies. Risks associated with produce consumption in the United States and the European Union have been linked to various factors such as irrigation water, cross-contamination, storage time and temperature abuse, infected food handlers, and unprocessed contaminated ingredients. The results of the current study indicate the complexity of produce products consumed across the globe and the difficulty in tracing illnesses back to specific food ingredients. [ABSTRACT FROM AUTHOR]

Although studies have quantified bacterial transfer between hands and various materials, cross-contamination between the surface of fresh citrus fruit and the edible portions during hand peeling has not been reported. This study quantifies transfer of Salmonella to the edible portion of citrus fruit from a contaminated peel during hand peeling. Citrus fruits used for this study were Citrus sinensis (sweet orange) cultivars 'Valencia' and 'Navel', Citrus unshiu (Satsuma mandarins), Citrus reticulata × Citrus paradisi ('Minneola' tangelo or 'Honeybell'), and C. paradisi (grapefruit) cultivar 'Marsh'. An avirulent Salmonella Typhimurium LT2 (ATCC 700720) resistant to rifampin was used for all experiments. The inoculum containing approximately 9 log CFU/mL (50 μL) was spot inoculated onto the equator, stem, or styler of each fruit and allowed to dry for 24 h. Six volunteers put on single-use latex gloves and peeled inoculated fruit. Peel, edible fruit portion, and gloves were collected and enumerated separately. Three replicates of the study were performed in which each volunteer peeled two inoculated fruit of each variety (n = 36 fruit per variety). Cross-contamination from contaminated surface of citrus fruits to edible portion or gloved hands during peeling was affected by inoculation sites. Average Salmonella transfer to the edible portion ranged from 0.16% (Valencia inoculated at the equator) to 5.41% (navel inoculated at the stem). Average Salmonella transfer to gloved hands ranged from 0.41% (grapefruit inoculated at the stem) to 8.97% (navel inoculated at the stem). Most Salmonella remained on the peel of citrus fruits. The average level of Salmonella remaining on the peel ranged from 5.37% (Minneola inoculated at the equator) to 66.3% (Satsuma inoculated at the styler). When grapefruit was inoculated, the Salmonella that remained on the peel showed a bimodal pattern in which some individuals left almost all Salmonella on the peel, while others left substantially less. [ABSTRACT FROM AUTHOR]

The microbial quality of water that comes into the edible portion of produce is believed to directly relate to the safety of produce, and metrics describing indicator organisms are commonly used to ensure safety. The US FDA Produce Safety Rule (PSR) sets very specific microbiological water quality metrics for agricultural water that contacts the harvestable portion of produce. Validation of these metrics for agricultural water is essential for produce safety. Water samples (500 mL) from six agricultural ponds were collected during the 2012/2013 and 2013/2014 growing seasons (46 and 44 samples respectively, 540 from all ponds). Microbial indicator populations (total coliforms, generic Escherichia coli, and enterococci) were enumerated, environmental variables (temperature, pH, conductivity, redox potential, and turbidity) measured, and pathogen presence evaluated by PCR. Salmonella isolates were serotyped and analyzed by pulsed-field gel electrophoresis. Following rain events, coliforms increased up to 4.2 log MPN/100 mL. Populations of coliforms and enterococci ranged from 2 to 8 and 1 to 5 log MPN/100 mL, respectively. Microbial indicators did not correlate with environmental variables, except pH (P<0.0001). The invA gene (Salmonella) was detected in 26/540 (4.8%) samples, in all ponds and growing seasons, and 14 serotypes detected. Six STEC genes were detected in samples: hly (83.3%), fliC (51.8%), eaeA (17.4%), rfbE (17.4%), stx-I (32.6%), stx-II (9.4%). While all ponds met the PSR requirements, at least one virulence gene from Salmonella (invA-4.8%) or STEC (stx-I-32.6%, stx-II-9.4%) was detected in each pond. Water quality for tested agricultural ponds, below recommended standards, did not guarantee the absence of pathogens. Investigating the relationships among physicochemical attributes, environmental factors, indicator microorganisms, and pathogen presence allows researchers to have a greater understanding of contamination risks from agricultural surface waters in the field. [ABSTRACT FROM AUTHOR]

Salmonella bacteria may internalize into tomato pulp when warm tomatoes from the field are submerged into colder water. Several washing steps may follow the initial washing and packing of tomatoes at the packinghouses; the potential for internalization into tomatoes in subsequent washing steps when tomatoes have a cooler pulp temperature is unknown. Our objective was to evaluate Salmonella internalization into mature green and red tomatoes with ambient (21°C) and refrigeration (4°C) pulp temperatures when they were submerged into water at various temperature differentials, simulating repacking and fresh-cut operations. Red (4°C and 21°C) and mature green (21°C) tomatoes were submerged (6 cm) into a six-strain Salmonella cocktail (6 log CFU/ml) and maintained at ±5 and 0°C temperature differentials for varying time intervals, ranging from 30 s to 5 min. Following submersion, tomatoes were surface sterilized using 70% ethanol, the stem abscission zone and blossom end epidermis were removed, and cores were recovered, separated into three segments, and analyzed. Salmonella populations in the segments were enumerated by most probable number (MPN) . The effects of temperature differential and maturity on Salmonella populations were analyzed; results were considered significant at a P value of ≥ 0.5. Internalized populations were not significantly different (P ≥ 0.5) across temperature differentials. Salmonella internalization was seen in tomatoes under all treatment conditions and was highest in the segment immediately below the stem abscission zone. However, populations were low (typically > 1 log MPN per segment) and varied greatly across temperature differentials. This suggests that the temperature differential between tomatoes and water beyond the initial packinghouse may be less important than submersion time in Salmonella internalization. [ABSTRACT FROM AUTHOR]

In-shell pecan samples (500 g) were collected over four harvest seasons (2010 to 2014) from seven pecan shelling facilities located in five U.S. states. Four varieties of pecans were analyzed: Mexican Improved, Native Seedlings, Southern Improved, and Western Improved. Pecan samples (100 g) were sent to a third party laboratory for initial Salmonella screening. When a sample was positive for Salmonella, the pathogen level was determined by the most-probable-number (MPN) method (25, 2.5, and 0.25 g). Two sample preparation strategies were used for the MPN analysis, and both strategies were combined for the reported MPN values. Forty-four (0.95%) of 4,641 in-shell pecan samples were positive for Salmonella during initial screening; prevalence by year was 0.47 to 1.4%. Prevalence was not significantly different between varieties: Mexican Improved, 1.2%; Native/Seedling, 0.99%; Southern Improved, 0.97%; and Western Improved, 0.75%. Salmonella was not isolated from 31 of 44 samples upon retesting during MPN analysis (<0.47 MPN/100 g). When Salmonella was detected, the levels were 0.47 to 39 MPN/100 g, with a mean of 2.4 MPN/100 g. Thirty-one Salmonella serotypes were obtained from 42 Salmonella-positive pecan samples; Enteritidis was the most common (12% of samples) followed by Javiana (9%) and Braenderup (7%). All Salmonella Enteritidis isolates were phage type 8. Pulsed-field gel electrophoresis analysis (Xbal) revealed within-serotype diversity, indicating introduction of contamination from a variety of sources. Most (64%) of the isolates were resistant to streptomycin or tetracycline, and 13% were resistant to three or more antibiotics. Salmonella prevalence and level on in-shell pecans is comparable to that on other nuts. [ABSTRACT FROM AUTHOR]

Poultry litter is applied to crop production land in the southern United States as a waste management strategy as it is a nitrogen-rich fertilizer and plentiful throughout the region. While litter is a known reservoir for human enteric pathogens including Salmonella enterica, little is known regarding pathogen prevalence, concentration, and common serotypes within the material. Litter from thirteen farms across four southern states was examined for Salmonella. Samples (n = 490) from six of the thirteen (46.2%) farms tested positive. Thirty-three samples out of 490 (6.7%) were Salmonella positive. Salmonella was ca. 95% less likely to be collected from stacked litter piles than from the poultry house floor or pasture, and every day increase in litter age reduced the likelihood of recovering Salmonella by 5.1%. When present, concentrations of Salmonella in contaminated poultry litter were variable, ranging from <0.45 to >280,000 MPN/g. The most prevalent serotypes found were Kentucky (45.5%), Kiambu (18.2%), and Michigan (12.1%). Salmonella Kentucky also had the greatest distribution and was found on 4 of the 6 (66.7%) positive farms. Results from this survey demonstrated that Salmonella prevalence and concentration in poultry litter is highly variable, and good agricultural practices are critical to safely use poultry litter as a soil amendment on fresh produce fields. [ABSTRACT FROM AUTHOR]

Strawberries and blueberries harvested at or near full-ripe maturity tend to be less firm and more susceptible to bruising during harvest and transport. The objective of this research was to determine the fate of Escherichia coli O157:H7 and Salmonella on bruised and intact surfaces of whole strawberries and blueberries at shipping (2°C) and retail display (15.5°C) temperatures. Strawberries and blueberries were either purchased from a supermarket or were harvested immediately prior to use; they were bruised using established protocols, were spot inoculated, and were incubated at 2 and 15.5°C. Strawberries, subjected to modified atmospheres, were further transferred to bags and were sealed in with an initial atmosphere of ca. 10% CO2 and 5% O2. Strawberries were sampled at 0, 2, 5, and 24 h and on days 3 and 7; blueberries were sampled on days 0, 1,3, and 7. After stomaching, samples were enumerated on nonselective and selective media, and populations were recorded as log CFU per berry. At both storage temperatures, population declines for both E. coli O157:H7 and Salmonella were seen under all conditions for strawberries. At 2 ± 2°C, E. coli O157:H7 and Salmonella populations on blueberries declined over 7 days under all conditions. At 15.5 ± 2°C, E. coli O157:H7 populations declined; however, Salmonella populations initially declined but increased to populations near or above initial populations over 7 days on blueberries. No overall significant differences were observed between bruised and intact treatments or between the two maturity levels for strawberries and blueberries. Modified atmospheric conditions did not affect the behavior of E. coli O157:H7 and Salmonella on strawberries at both temperatures. This research indicates that E. coli O157:H7 and Salmonella do not grow on strawberries at shipping or retail display temperatures, even when they are harvested at a maturity prone to bruising; however, Salmonella growth may occur on bruised full ripe blueberries under retail display temperatures. [ABSTRACT FROM AUTHOR]

Florida Tomato Good Agricultural Practices (T-GAPs) mandate the removal of dirt and debris from tomatoes during harvest but do not provide any specific regulations or guidance; thus, the current practice of using cloths needs to be evaluated. This study examined Salmonella transfer from inoculated green tomatoes to uninoculated cloths and from inoculated cloths to uninoculated tomatoes, upon single and multiple touches. Tomatoes were spot inoculated with a rifampin-resistant Salmonella cocktail (107 CFU per tomato) and were touched with cloth (clean, dirty-dry, dirty-wet) at 0, 1, or 24 h postinoculalion. Salmonella was enumerated on tryptic soy agar, followed by enrichments when necessary. The transfer direction was then reversed by touching freshly inoculated cloths with uninoculated tomatoes. Transfer coefficients (TCs) were then calculated. Salmonella TCs from inoculated tomato and cloth were highest when the inoculum was wet (0.44 ± 0.13 to 0.32 ± 0.12), regardless of the condition of the cloth. Although Salmonella TCs from inoculated tomato to uninoculated cloth decreased significantly when the inoculum was dried (0.17 ± 0.23 to 0.01 ± 0.00), low levels of Salmonella were detected on cloth even after 24 h of drying. Inoculated dirty cloth did not transfer more Salmonella compared with inoculated clean cloth, and Salmonella survival was not higher on dirty cloth. When inoculated clean cloth (wet) was touched with 25 tomatoes, significantly higher levels of Salmonella were transferred to the first, second, and fourth tomatoes (0.03 ± 0.10 to 0.09 ± 0.02). However, inoculated dirty-wet (below limit of detection) and dirty-dry (0.00 to 0.04 ± 0.01) cloths transferred similar levels of Salmonella to all 25 tomatoes. Results indicate a low risk of potential Salmonella contamination when the same cloth is used multiple times for debris removal, especially under high moisture levels. Results also show that the use of dirty cloths did not increase the risk of Salmonella cross-contamination. [ABSTRACT FROM AUTHOR]

Human norovirus (HuNoV) is the leading cause of foodborne illness outbreaks and the second most common cause of waterborne infections in the United States. The goal of this research was to investigate the antiviral activity of chitosan microparticles (CMs) against HuNoV GII.4 Sydney and its cultivable surrogate Tulane virus (TuV) in suspensions mimicking fecally contaminated water. CMs were prepared by cross-linking chitosan molecules with sodium sulfate, and the antiviral activity of CMs was assessed with an infectivity assay on TuV and by quantitative reverse transcription PCR on TuV and HuNoV. A 3% CM suspension in phosphate-buffered saline (pH 7.2) bound to TuV particles but had a negligible impact on virus infectivity (P > 0.05). A 10-min contact time resulted in a 1.5-log reduction in genomic copies per mL of TuV and HuNoV in fecal suspensions (P < 0.05). Despite the negligible impact on viral infectivity, CMs can moderately bind to infectious virus particles and help purify environmental water by removing these particles. In this study, TuV was a suitable surrogate for HuNoV with similar log reductions in fecal suspension. These findings highlight the potential application of CM as a novel treatment to minimize the spread of waterborne viral pathogens. CMs had antiviral activity against TuV and HuNoV in fecal suspensions. CMs impact on TuV infectivity was negligible. CMs bound to virus particles regardless of the fecal load. CMs have the potential to remove HuNoV from water. [ABSTRACT FROM AUTHOR]

Many outbreaks of foodborne illness associated with the consumption of fresh-cut melons have been reported. The objective of our research was to develop a mathematical model that predicts the growth rate of Salmonella on fresh-cut cantaloupe over a range of storage temperatures and to validate that model by using Salmonella and Escherichia coli O157:H7 on cantaloupe, honeydew, and watermelon, using both new data and data from the published studies. The growth of Salmonella on honeydew and watermelon and E. coli O157:H7 on cantaloupe, honeydew, and watermelon was monitored at temperatures of 4 to 25°C. The Ratkowsky (or square-root model) was used to describe Salmonella growth on cantaloupe as a function of storage temperature. Our results show that the levels of Salmonella on fresh-cut cantaloupe with an initial load of 3 log CFU/g can reach over 7 log CFU/g at 25°C within 24 h. No growth was observed at 4°C. A linear correlation was observed between the square root of Salmonella growth rate and temperature, such that √growth rate = 0.026 x (T - 5.613), R² = 0.9779. The model was generally suitable for predicting the growth of both Salmonella and E. coli O157:H7 on cantaloupe, honeydew, and watermelon, for both new data and data from the published literature. When compared with existing models for growth of Salmonella, the new model predicts a theoretic minimum growth temperature similar to the ComBase Predictive Models and Pathogen Modeling Program models but lower than other food-specific models. The ComBase Prediction Models results are very similar to the model developed in this study. Our research confirms that Salmonella can grow quickly and reach high concentrations when cut cantaloupe is stored at ambient temperatures, without visual signs of spoilage. Our model provides a fast and cost-effective method to estimate the effects of storage temperature on fresh-cut melon safety and could also be used in subsequent quantitative microbial risk assessments. [ABSTRACT FROM AUTHOR]

Abstract: Salmonella enterica strains with rdar (red dry and rough) and saw (smooth and white) morphotypes have previously been associated with tomato outbreaks but the dispersal mechanisms of these morphotypes are still poorly understood. In this study, Salmonella adhesion was distinguished from attachment by comparing different contact periods. Initial adhesion of rdar and saw morphotypes of Salmonella was compared in relation to tomato plants with different leaf trichome densities. Trichome densities were increased or reduced by treatment with jasmonic or salicylic acid, respectively. The overall effect of Salmonella morphotype and trichome density on splash dispersal was assessed in a rain simulator and correlated to cell hydrophobicity and initial adhesion. The presence of the rdar morphotype increased initial adhesion at high trichome densities but not at low trichome densities. Attachment of the rdar strain occurred after 30s contact time regardless of trichome density. Splash dispersal was slightly further for the saw morphotype than the rdar morphotype of S. enterica at all trichome densities. Salmonella cells of both morphotypes survived significantly better on the surface of high trichome density leaflets. [Copyright &y& Elsevier]

Abstract: Outbreaks of Salmonella enterica have been associated with tomatoes and traced back to production areas but the spread of Salmonella in agricultural fields is still poorly understood. Post-rain Salmonella transfer from a point source to the air and then to tomato plants was evaluated. GFP-labeled kanamycin-resistant S. enterica serovar Typhimurium (108 CFU/mL) with and without expression of the rdar morphotype (rough colonies; cells with fimbriae and cellulose) was used as the point source in the center of a rain simulator. Rain intensities of 60 and 110mm/h were applied for 5, 10, 20, and 30min. Petri dishes with lactose broth and tomato plants with fruit (50–80cm high) were placed in the simulator after the rain had ceased. Salmonella recovery from air was maximum (300CFU/plate) after a rain episode of 60mm/h for 10min at distances of at least 85.5cm above the source and when the rdar morphotype strain was used. Small scale experiments showed that the smooth-colony strain without fimbriae precipitated from the air in significantly higher numbers than the rdar strain. Transfer of aerial Salmonella with the rdar morphotype to tomato fruits on plants followed a beta distribution (2.5950, 4.7393) within the generalized range from 0 to 30min of rain. Results show for the first time that Salmonella may transfer from rain to the air and contaminate tomato fruits at levels that could possibly be infectious to humans. [Copyright &y& Elsevier]

Modified Moore swabs (MMS; consisting of a polyvinyl chloride cartridge filled with gauze) capture microorganisms within the packed gauze as water flows through the cartridge, while continuous flow centrifugation (CFC) uses centrifugation to sediment the microorganisms while water continuously flows in the system. This study evaluated and compared the efficacy of MMS and CFC for concentration and subsequent detection of Escherichia coli O157:H7 and Salmonella from large volumes of water (10 liters). Water samples were spiked at levels of 10¹, 10², 10³, and 104; CFU/100 ml with three-strain cocktails of either E. coli O157:H7 or Salmonella serovars, which had been previously transformed with a plasmid to express resistance to ampicillin as well as green, red, or cyan fluorescent proteins. Plating was performed before and after concentration on tryptic soy agar supplemented with ampicillin in order to quantitate the concentration efficiencies of each method. The two lowest spiking levels were also enriched in low volumes of tryptic soy broth supplemented with ampicillin followed by testing via lateral flow devices. Significant (P < 0.05) concentrations of initial levels of E. coli O157:H7 in the range of 0.7 to 1.0 and 1.2 to 1.4 log were achieved within approximately 35 min of processing time via MMS and CFC, respectively. Similarly, significant (P < 0.05) concentrations were also achieved for Salmonella with 0.9 to 1.2 and 1.2 to 1.4 log concentration for MMS and CFC, respectively. There were no statistical differences (P > 0.05) between the two concentration methods in their ability to concentrate either of the two target bacteria. Significantly (P < 0.05) more spiked samples were detected by lateral flow devices following concentration and enrichment than for nonconcentrated, enriched samples. It is concluded that both MMS and CFC have potential to be used to enhance the sensitivity of downstream bacterial detection methods used to test irrigation water for the presence of foodborne pathogens. [ABSTRACT FROM AUTHOR]

There are approximately 140 million tons of over 3,000 types of tropical fruits produced annually worldwide. Tropical fruits, once unfamiliar and rare to the temperate market, are now gaining widespread acceptance. Tropical fruits are found in a variety of forms, including whole, fresh cut, dried, juice blends, frozen, pulp, and nectars in markets around the world. Documented outbreaks of foodborne disease associated with tropical fruits have occurred. Norovirus and Salmonella are the leading viral and bacterial pathogens, respectively, documented to have caused outbreaks of infections associated with consumption of tropical fruits. Sources of contamination of tropical fruit have been identified in the production environment and postharvest handling, primarily related to sanitation issues. Limited data exist on the specific route of transmission from these sources. Research on the microbial safety of tropical fruits is minimal; with the growing market for tropical fruit expected to increase by 33% in 2010 this research area needs to be addressed. The aim of this review is to discuss the foodborne pathogen outbreaks associated tropical fruit consumption, research previously completed on pathogen behavior on tropical fruits, preventive strategies for pathogen contamination, and research needs. [ABSTRACT FROM AUTHOR]

Abstract: Alicyclobacillus spp. is an important thermoacidophilic, spore-forming spoilage bacterium that is a major concern for beverage and juice industries. In order to develop effective control strategies and adequately address the prevalence of contamination sources, it is necessary to characterize Alicyclobacillus'' ecology in fruit, juice and beverage production and processing environments. Alicyclobacillus spp. isolates were collected from juice, beverage, ingredients, and environmental samples over a period of ten years. A total of 141 isolates were characterized as Alicyclobacillus spp. by 16S rRNA analysis and the most frequently isolated species was found to be Alicyclobacillus acidoterrestris (45%), A. acidocaldarius subsp. acidocaldarius (30%), and A. acidocaldarius (11%). The majority of thermotolerant sporeformers isolated from apple juices and concentrates was found to be A. acidoterrestris (24 out of 36 total apple isolates); while A. acidoterrestris was most frequently associated with citrus, citrus concentrates, and their associated environments, isolated by University of Florida (UF) (15 out of total 28 UF citrus isolates). However, A. acidocaldarius and subsp. acidocaldarius were frequently isolated by Cornell University (CU) (29 out of 35 CU citrus isolates), from citrus juices made from concentrate. Four major haplotypes of Alicyclobacillus spp. were identified based on the 16S rRNA gene sequencing from the 141 isolates tested. The Allelic Types (ATs) matched the phylogenetic analysis grouping of the different Alicyclobacillus spp. based on the isolation source. Our results suggest a predisposition for certain ATs of Alicyclobacillus spp. depending on juice or ingredient isolation source. [Copyright &y& Elsevier]

The purpose of this study was to evaluate the performance of artificial intelligence tools for the prediction of Salmonella presence and absence in agricultural surface waters based on the population of microbiological indicators (total coliform, generic Escherichia coli, and enterococci) and physicochemical attributes of water (air and water temperature, conductivity, ORP, pH, and turbidity). Previously collected data set from six agricultural ponds monitored for two growing seasons were used for analysis. Classification algorithms including artificial neural networks (ANNs), the nearest neighborhood algorithm (kNN), and support vector machines (SVM) were trained and tested with a 539‐point data set for optimum prediction accuracy. Classification accuracy performances were validated with data set (400 samples) collected from different agricultural surface water sources. All tested algorithms yielded the highest accuracy around 75 ± 1% for generic E. coli followed by enterococci (65 ± 5%) and total coliform (60 ± 10%). Classifiers calculated 6–15% higher accuracy ranging from 62 to 66% for turbidity than all other tested physicochemical attributes. Based on E. coli populations measured in other water sources, trained algorithms predicted the presence and absence of Salmonella with an accuracy between 58.15 and 59.23%. The classification performance of ANN, kNN, and SVM algorithms are encouraging for the prediction of Salmonella in agricultural surface waters. [ABSTRACT FROM AUTHOR]

Heightened concerns about wildlife on produce farms and possible introduction of pathogens to the food supply have resulted in required actions following intrusion events. The purpose of this study was to evaluate the survival of Salmonella in feces from cattle and various wild animals (feral pigs, waterfowl, deer, and raccoons) in California, Delaware, Florida, and Ohio. Feces were inoculated with rifampin-resistant Salmonella enterica cocktails that included six serotypes: Typhimurium, Montevideo, Anatum, Javiana, Braenderup, and Newport (104 to 106 CFU/g). Fecal samples were stored at ambient temperature. Populations were enumerated for up to 1 year (364 days) by spread plating onto tryptic soy agar supplemented with rifampin. When no colonies were detected, samples were enriched. Colonies were banked on various sampling days based on availability of serotyping in each state. During the 364-day storage period, Salmonella populations decreased to ≤2.0 log CFU/g by day 84 in pig, waterfowl, and raccoon feces from all states. Salmonella populations in cattle and deer feces were 3.3 to 6.1 log CFU/g on day 336 or 364; however, in Ohio Salmonella was not detected after 120 days. Salmonella serotypes Anatum, Braenderup, and Javiana were the predominant serotypes throughout the storage period in all animal feces and states. Determination of appropriate risk mitigation strategies following animal intrusions can improve our understanding of pathogen survival in animal feces. Survival of Salmonella in fecal samples differed among animal types. Salmonella survived for at least 1 year in cattle and deer feces. Salmonella Javiana was the predominant serotype among recovered cells. Responses to wild animal intrusions in produce fields may need to be region specific. [ABSTRACT FROM AUTHOR]