Primers specific for the fimbrial major subunit gene stdA can be used to detect Salmonella enterica serovars

The feasibility of using two primers internal to the stdA gene (which encodes the fimbrial major subunit of the std fimbrial gene cluster in Salmonella enterica serovar Typhi) to detect Salmonella by PCR was explored. The 518-bp stdA specific sequence was conserved among 268 strains from 45 serovars of S. enterica. One Salmonella bongori CCUG 30042 strain and 34 non-Salmonella strains did not possess this sequence. A sensitivity test revealed that the stdA-specific primer set detected 3.4 x 10(-1) pg of genomic DNA and 3.0 x 10(5) CFU/ml with serial dilutions of Salmonella Typhimurium cells. In vitro testing for specificity using pig carcass sponge samples contaminated with Salmonella Typhimurium also was performed. An initial Salmonella Typhimurium inoculum of 4.4 x 10(1) CFU/ml in pig carcass exudates reached the stdA primer detection level after preenrichment in buffered peptone water at 37 degrees C for 18 h in the presence of indigenous non-Salmonella flora at 4.0 X 10(7) CFU/ml, but the detection level decreased to 4.4 x 10(0) CFU/ml after selective enrichment in Rappaport-Vassiliadis R10 broth for 18 h at 42 degrees C. The PCR method with primers specific for stdA is a quick and sensitive tool for detecting S. enterica, which is an important cause of foodborne disease.     

Radiation processing for elimination of Salmonella typhimurium from inoculated seeds used for sprout making in India and effect of irradiation on germination of seeds

The effect of radiation processing on the germination of the sprout seeds mung (Phaseolus aureus), matki (Phaseolus aconitifolius), chana (Cicer arietinum), and vatana (Pisum sativum) in terms of percent germination, germination yield, sprout length, vitamin C content, and texture was investigated. Gradual decreases in the percent germination, germination yield, and sprout length with increases in radiation dose (0.5 to 2.0 kGy) were observed. Vitamin C content and texture remained unaffected for the seeds treated with doses of up to 2 kGy. To determine the efficacy of radiation treatment in elimination of foodborne pathogens, seeds inoculated with 4 log CFU/g of Salmonella Typhimurium were treated with radiation doses of 1 and 2 kGy. A reduction in counts of Salmonella Typhimurium in inoculated seeds after radiation treatment was observed. A radiation dose of 2 kGy resulted in the complete elimination of 4 log CFU/g of Salmonella Typhimurium from the inoculated seeds. However, on sprouting for 48 h, the count of Salmonella Typhimurium reached 8 log CFU/g for the control seeds and the seeds treated with a 1-kGy radiation dose. The aerobic plate counts for seeds were 2.0 to 2.6 log CFU/g, which were reduced to 0.9 to 1.2 log CFU/g on treatment with a 2-kGy radiation dose. On sprouting for 48 h, the aerobic plate count reached 8 log CFU/g for both the control and radiation-treated seeds. The study demonstrates that irradiation can control bacterial levels on seeds but not contamination introduced during posttreatment handling. Therefore, radiation processing of the final product (sprouts) is recommended, rather than of the seeds.     

Fate of Staphylococcus aureus, Salmonella enterica serovar typhimurium, and vibrio vulnificus in raw oysters treated with chitosan

The fate of Staphylococcus aureus, Salmonella enterica serovar Typhimurium, and Vibrio vulnificus in oysters treated with chitosan was investigated. Three concentrations (0.5, 1.0, and 2.0%) of chitosan in 0.5% hydrochloric acid were prepared and coated onto raw oysters, which were then stored at 4 degrees C for 12 days. Untreated oysters and oysters coated with 0.5% hydrochloric acid without chitosan were used as controls. S. aureus cells were most sensitive to 2.0% chitosan followed by 0.5 and 1.0%. In general, chitosan treatment of oysters produced a decline in the population of S. aureus by 1 to 4 log CFU/ml compared with the untreated control. Chitosan treatment had no influence on the reduction of Salmonella Typhimurium over the 12-day storage period; inhibition of Salmonella Typhimurium growth was similar in both the control samples and the chitosan-treated samples. However, time of storage had a major effect on the survival of Salmonella Typhimurium on oysters. Neither time nor chitosan concentration had a significant effect on the growth of V. vulnificus during storage. All treatments were similar in inhibiting V. vulnificus growth.     

Survival and growth of foodborne pathogens during cooking and storage of oriental-style rice cakes

Fresh cooked rice cakes for retail sale are typically held at room temperature because refrigeration dramatically reduces their quality. Room temperature, high water activity, and a pH of > 4.6 provided an environment conducive to pathogen growth. To date, no studies have been published regarding survival and growth of foodborne pathogens in fresh cooked rice cakes. This study was undertaken to investigate the effect of steam cooking on foodborne pathogens and their subsequent growth in five varieties of rice cakes made from flours of regular rice, sweet rice, white rice, tapioca, and mung bean. Bacillus cereus spores were detected in white rice, tapioca, and mung bean samples. The rice cake flours were inoculated with non-spore-forming foodborne pathogens (Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Listeria monocytogenes, and Staphylococcus aureus) or spore-forming bacteria (Bacillus cereus) and steam cooked (100 degrees C) for 30 min. Steam cooking significantly reduced (> 6 log CFU/g) non-spore-forming foodborne pathogens in all samples and inactivated spores of B. cereus by 1 to 2 log CFU/g. Although spores of B. cereus survived steam cooking and germinated during 3 days of storage at room temperature, populations in most rice cakes remained below 106 CFU/g, which is the threshold for producing toxin. Rice cakes made from mung bean flour supported growth and germination of B. cereus spores above that critical level. In mung bean rice cakes, enterotoxin production was detected by the second day, when B cereus cell populations reached about 6.9 log CFU/g. The toxin concentration increased with storage time. However, our results suggest that rapid growth of total mesophilic microorganisms by more than 7 to 8 log CFU/ml during the first day of storage produced off flavors and spoilage before B. cereus was able to grow enough to produce toxins. Therefore, steam-cooked rice cakes made from a variety of flours including mung bean flour are safe for sale for up to 1 day after storage at room temperature and are free of B. cereus toxins.     

Assessment of contamination potential of lettuce by Salmonella enterica serovar Newport added to the plant growing medium

The capacity of Salmonella enterica serovar Newport to contaminate Romaine lettuce (Lactuca sativa L. cv. Nogal) via the root system was evaluated in 17-, 20-, and 33-day-old plants. Apparent internalization of Salmonella via the root to the above-ground parts was identified in 33- but not 17- or 20-day-old plants and was stimulated by root decapitation. Leaves of lettuce plants with intact and damaged roots harbored Salmonella at 500 +/- 120 and 5,130 +/- 440 CFU/g of leaf, respectively, at 2 days postinoculation but not 5 days later. These findings are first to suggest that Salmonella Newport can translocate from contaminated roots to the aerial parts of lettuce seedlings and propose that the process is dependent on the developmental stage of the plant.     

Competitive inhibition bacteria of bovine origin against Salmonella serovars

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 Serratia fonticola, 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 degrees C in cattle feces. B. circulans (10(5) CFU/g of inoculum) significantly reduced Salmonella Newport on days 3 and 5 and on day 21 with 10(8) CFU/g of inoculum at 37 degrees C. At 21degrees C, significant reductions of Salmonella Typhimurium DT104 occurred with 10(8) CFU of gram-negative competitive inhibition bacteria per g and 10(5) CFU of B. circulans per g on day 5 only. No significant reductions were observed with Salmonella Newport at 21 degrees 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.     

Disinfection of mung bean seed with gaseous acetic acid.

Mung bean seed inoculated with Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes (3 to 5 log CFU/g) was exposed to gaseous acetic acid in an aluminum fumigation chamber. Salmonella Typhimurium and E. coli O157:H7 were not detected by enrichment of seeds treated with 242 microl of acetic acid per liter of air for 12 h at 45 degrees C. L. monocytogenes was recovered by enrichment from two of 10 25-g seed samples treated in this manner. Fumigation with gaseous acetic acid was also lethal to indigenous bacteria and fungi on mung bean seed. The treatment did not significantly reduce seed germination rates, and no differences in surface microstructure were observed between treated and untreated seed viewed by scanning electron microscopy.     

INTERACTIONS BETWEEN SALMONELLA ENTERICA SUBSPECIES ENTERICA SEROVAR TYPHIMURIUM AND COWPEA (VIGNA UNGUICULATA VARIETY SINENSIS) SEEDS, PLANTS AND PERSISTENCE IN HAY

Dynamics of persistence of Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium), a common food‐ and waterborne zoonotic serovar, on cowpea (Vigna unguiculata variety sinensis), a fodder and green vegetable plant, were studied. The findings revealed that S. Typhimurium not only reduced germination of cowpea seed (P < 0.01), but also caused defects in sprouts (P < 0.02). S. Typhimurium inoculation on seeds before sowing had a more pronounced effect (P < 0.01; i.e., loss in germination and appearance of defective sprouts) than sowing seeds in S. Typhimurium‐inoculated soil. S. Typhimurium persisted in saplings and adult plants up to 45 days of plant age and up to 60 days in hay. The cowpea plants grown in sterile Salmonella‐free soil did not support colonization of S. Typhimurium in different parts. A reduction in the population of Salmonella appeared as early as on the fifth day and decreased with advancing plant age. At 21 days of age, the cowpea plants had no Salmonella in their aerial parts and were free of the pathogen within 3 h of inoculation. Salmonella persisted in stumps of the plants throughout the observation, irrespective of age of the plants at the time of inoculation. The study revealed the persistence and the phytopathogenic potential of Salmonella on cowpea.     

Microbiological evaluation of sprouts marketed in Mumbai, India, and its suburbs

A study was undertaken to assess the microbiological quality of sprouts marketed in Mumbai and its suburbs. A total of 124 sprout samples of four different legumes--mung (Phaseolus aureus), matki (Phaseolus aconitifolius), chana (Cicer arietinum), and vatana (Pisum sativum)--were analyzed over a period of 12 months for aerobic plate counts, coliforms, yeast and mold counts, staphylococci, Salmonella, Listeria monocytogenes, Escherichia coli, E. coli O157:H7, and coagulase-positive Staphylococcus aureus. Aerobic plate counts ranged from 7.6 to 8.9 log CFU/g, coliform counts ranged from 5.4 to 7.9 log CFU/g, yeast and mold counts ranged from 3.6 to 7.3 log CFU/g, and staphylococci counts ranged from 3.3 to 6.6 log CFU/ g. Nonpathogenic E. coli was detected in 13% of the mung, 26% of the matki, 40% of the chana, and 19% of the vatana samples. Salmonella Typhimurium was detected in 21% of the mung, 40% of the matki, and 4% of the chana samples. Salmonella Dublin was detected in 2% of the mung samples, and Salmonella Washington was detected in 4% of the matki samples. L. monocytogenes and E. coli O157:H7 were not detected in any of the samples examined. Coagulase-positive S. aureus was detected in 4% of the mung, 11% of the matki, and 4% of the chana samples. The results indicated that the marketed sprouts were of poor microbiological quality; therefore, further processing, such as radiation processing, is needed to ensure their safety.     

Identification of Salmonella enterica serovar Typhimurium using specific PCR primers obtained by comparative genomics in Salmonella serovars

Salmonella enterica serovar Typhimurium is a major foodborne pathogen throughout the world. Until now, the specific target genes for the detection and identification of serovar Typhimurium have not been developed. To determine the specific probes for serovar Typhimurium, the genes of serovar Typhimurium LT2 that were expected to be unique were selected with the BLAST (Basic Local Alignment Search Tool) program within GenBank. The selected genes were compared with 11 genomic sequences of various Salmonella serovars by BLAST. Of these selected genes, 10 were expected to be specific to serovar Typhimurium and were not related to virulence factor genes of Salmonella pathogenicity island or to genes of the O and H antigens of Salmonella. Primers for the 10 selected genes were constructed, and PCRs were evaluated with various genomic DNAs of Salmonella and non-Salmonella strains for the specific identification of Salmonella serovar Typhimurium. Among all the primer sets for the 10 genes, STM4497 showed the highest degree of specificity to serovar Typhimurium. In this study, a specific primer set for Salmonella serovar Typhimurium was developed on the basis of the comparison of genomic sequences between Salmonella serovars and was validated with PCR. This method of comparative genomics to select target genes or sequences can be applied to the specific detection of microorganisms.     

Development of PCR primers for the detection of Salmonella enterica serovar Choleraesuis based on the fliC gene

Salmonella enterica serovar Choleraesuis may cause swine salmonellosis and human infection. Because the conventional method for detection of this Salmonella serovar may take 3 to 5 days, a PCR method for detection was evaluated. By comparing the sequence of the phase 1 flagellin (fliC) gene of Salmonella Choleraesuis with that of other Salmonella serovars and of other bacteria species available in GenBank, two PCR primers (flinC-F and flinC-R) were designed. Using these primers, all 97 Salmonella Choleraesuis strains assayed generated the expected PCR product, with a molecular mass of 963 bp. Except for S. enterica Paratyphi C, Salmonella isolates other than Salmonella Choleraesuis and non-Salmonella isolates, including strains of Enterobacteriaceae, all generated negative PCR results. Salmonella Paratyphi C could be differentiated from Salmonella Choleraesuis through the use of primers designed from the viaB gene. When Salmonella Choleraesuis isolates from swine stool, pork, liver, feed, and human whole blood samples were assayed with a preenrichment step, as low as 1 CFU/g or ml of the original sample could be detected.     

Quantification of contamination of lettuce by GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium

The primary objective of this study was to determine the possibility of internalization of GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium (S. Typhimurium) strains MAE 110 (multi-cellular morphology) and 119 (wild type morphology) into lettuce seedlings (Lactuca sativa cv. Tamburo) grown in an inoculated hydroponic and soil system. The second aim was to quantify the level of contamination with the use of a proper surface sterilization method. Silver nitrate was superior in reducing the number of viable bacteria on leave surfaces compared to sodium hypochlorite and ethanol. With the hydroponic system internal colonization of lettuce only occurred at high densities with S. Typhimurium MAE 119. With the soil system E. coli O157:H7, S. Typhimurium 110 and S. Typhimurium 119 were found at considerable densities in sterilized leaf samples (respectively, 3.95, 2.57 and 2.37 log cfu/g on average) with prevalences of 0.29, 0.23 and 0.15, respectively. No statistical differences were observed between the Salmonella strains. A negative correlation was observed between shoot weight and leaf contamination. The observed presence of the pathogens in lettuce, after thorough surface sterilization, demonstrates the possible presence of human pathogens in locations were they are unlikely to be removed by the actions of consumer washing and therefore pose a serious threat when occurring in field situations.     

Inhibition of Salmonella Typhimurium and Listeria monocytogenes in mung bean sprouts by chemical treatment

This study was undertaken to compare the efficacies of chlorous acid (268 ppm), sodium hypochlorite (200 ppm), and lactic acid (2%) in eliminating total mesophilic microorganisms, Salmonella Typhimurium, and Listeria monocytogenes on commercial mung bean sprouts immediately after treatment and during posttreatment refrigerated storage. Treatment with sodium hypochlorite for 10 min did not reduce the total aerobic count. However, treatment with lactic acid and chlorous acid for 10 min initially reduced the total aerobic count by 0.6 and 0.8 log CFU/g, respectively, and maintained the same level or a lower level of the total aerobic count during the storage time. Treatment with chlorous acid reduced Salmonella Typhimurium from 5.0 log to undetectable levels (<0.48 log CFU/g), and the pathogen remained undetectable over a 9-day storage period. Treatment with lactic acid resulted in an initial 3-log reduction and further reduced the number of Salmonella Typhimurium cells to undetectable levels after 3 days. For L. monocytogenes, treatment with chlorous acid resulted in an initial 5-log reduction, and treatment with lactic acid resulted in a 2-log reduction at the beginning and undetectable levels after 9 days. When chemically injured cells were investigated by the selective overlay method, no statistical difference was observed (P < 0.05) between the number of injured cells recovered following treatment with chlorous acid and the number of bacteria counted on selective media, whereas sodium hypochlorite generated more injured cells than the other treatments did. These data suggest that treatment with chlorous acid may be useful in reducing total mesophilic microorganisms, Salmonella Typhimurium, and L. monocytogenes in commercial mung bean sprouts.     

Inactivation of Escherichia coli O157:H7, Salmonella typhimurium DT104, and Listeria monocytogenes on inoculated alfalfa seeds with a fatty acid-based sanitizer

Alfalfa seeds were inoculated with a three-strain cocktail of Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium DT104, or Listeria monocytogenes by immersion to contain approximately 6 to 8 log CFU/g and then treated with a fatty acid-based sanitizer containing 250 ppm of peroxyacid, 1,000 ppm of caprylic and capric acids (Emery 658), 1,000 ppm of lactic acid, and 500 ppm of glycerol monolaurate at a reference concentration of 1X. Inoculated seeds were immersed at sanitizer concentrations of 5X, 10X, and 15X for 1, 3, 5, and 10 min and then assessed for pathogen survivors by direct plating. The lowest concentration that decreased all three pathogens by >5 log was 15. After a 3-min exposure to the 15X concentration, populations of E. coli O157:H7, Salmonella Typhimurium DT104, and L. monocytogenes decreased by >5.45, >5.62, and >6.92 log, respectively, with no sublethal injury and no significant loss in seed germination rate or final sprout yield. The components of this 15x concentration (treatment A) were assessed independently and in various combinations to optimize antimicrobial activity. With inoculated seeds, treatment C (15,000 ppm of Emery 658, 15,000 ppm of lactic acid, and 7,500 ppm of glycerol monolaurate) decreased Salmonella Typhimurium, E. coli O157:H7, and L. monocytogenes by 6.23 and 5.57 log, 4.77 and 6.29 log, and 3.86 and 4.21 log after 3 and 5 min of exposure, respectively. Treatment D (15,000 ppm of Emery 658 and 15,000 ppm of lactic acid) reduced Salmonella Typhimurium by >6.90 log regardless of exposure time and E. coli )157:H7 and L. monocytogenes by 4.60 and >5.18 log and 3.55 and 3.14 log after 3 and 5 min, respectively. No significant differences (P > 0.05) were found between treatments A, C, and D. Overall, treatment D, which contained Emery 658 and lactic acid as active ingredients, reduced E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes populations by 3.55 to >6.90 log and may provide a viable alternative to the recommended 20,000 ppm of chlorine for sanitizing alfalfa seeds.     

Optimization and validation of a simple method using P22::luxAB bacteriophage for rapid detection of Salmonella enterica serotypes A, B, and D in poultry samples

A simple method was developed for the fast and inexpensive detection of Salmonella Typhimurium using a recombinant P22::luxAB phage. All the steps from phage production to detection were considered. A strain of Salmonella Typhimurium harboring the prophage P22::luxAB was grown in batch culture to produce spontaneously the recombinant bacteriophage. Batch production to stationary phase was better for propagation of the phage and led to a total population of 4.3 x 10(9) (+/-4.3 x 10(9)) PFU/ml of P22, including only 1.4 x 10(6) (+/-1 x 10(6)) PFU/ml harboring the luxAB genes. After preenrichment, a simple four-step bioassay was tested and optimized for several parameters. The detection limit of the luminometer was only 5 x 10(2) (+/-1.75 x 10(2)) CFU Salmonella Typhimurium per ml, but increased to 1.5 x 10(4) (+/-1.17 x 10(4)) CFU Salmonella Typhimurium per ml when the cells were in a complex matrix. The detection limit after the preenrichment was 6.5 x 10(3) (+/-1.5 x 10(3)) CFU Salmonella Typhimurium per ml, but the detection limit after the preenrichment also increased markedly to 1.65 x 10(5) (+/-0.15 x 10(5)) CFU Salmonella Typhimurium per ml when Salmonella Typhimurium was in a complex matrix. Finally, the bioassay was applied to the detection of Salmonella Typhimurium LT2 in 14 different feed and environmental samples (including duck feed, litters, and feces) spiked either before or after the preenrichment process. It was possible to detect Salmonella Typhimurium LT2 in all samples within 16 h.     

Dispersal of Salmonella Typhimurium by rain splash onto tomato plants

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 10? 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.     

Effectiveness of radiation processing in elimination of Salmonella typhimurium and Listeria monocytogenes from sprouts

The effectiveness of radiation treatment in eliminating Salmonella Typhimurium and Listeria monocytogenes on laboratory inoculated ready-to-eat sprouts was studied. Decimal reduction doses (D10-values) for Salmonella Typhimurium and L. monocytogenes in dry seeds of mung (green gram), matki (dew gram), chana (chick pea), and vatana (garden pea) ranged from 0.189 to 0.303 kGy and 0.294 to 0.344 kGy, respectively. In sprouts made from these seeds, the D10-values ranged from 0.192 to 0.208 kGy for Salmonella Typhimurium and from 0.526 to 0.588 kGy for L. monocytogenes. Radiation treatment with a 2-kGy dose resulted in complete elimination of 10(4) CFU/g of Salmonella Typhimurium and 10(3) CFU/g of L. monocytogenes from all the four varieties of sprouts. No recovery of Salmonella Typhimurium and L. monocytogenes was observed in the radiation treated samples stored at 4 and 8 degrees C up to 12 days. Radiation treatment with 1 kGy and 2 kGy resulted in a reduction of aerobic plate counts and coliform counts by 2 and 4 log CFU/g, respectively; the yeast and mold counts and staphylococci counts decreased by 1 and 2 log CFU/g, respectively. However, during postirradiation storage at 4 and 8 degrees C, aerobic plate counts, coliform counts, yeast and mold counts, and staphylococci counts remained constant throughout the incubation period. This study demonstrates that a 2-kGy dose of irradiation could be an effective method of processing to ensure microbial safety of sprouts.     

Salmonella can reach tomato fruits on plants exposed to aerosols formed by rain

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 (10(8)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.     

Survival of Campylobacter jejuni and Salmonella enterica Typhimurium in vacuum-packed, moisture-enhanced pork

The abilities of Campylobacter jejuni and Salmonella enterica Typhimurium to survive in vacuum-packaged, moisture-enhanced pork stored at 4 or 10°C were examined. Pork loins were surface inoculated with either C. jejuni or Salmonella Typhimurium and then moisture enhanced to a target of 10 or 20%. The enhanced pork loins were sliced 1 cm thick and vacuum packaged. A pork loin without moisture enhancement was sliced and vacuum packaged as a control. Samples were collected, plated, and the numbers of surviving organisms were determined periodically during storage at 4 and 10°C. The numbers of C. jejuni or Salmonella Typhimurium in samples with different moisture enhancement levels were similar (P > 0.05). No significant differences (P > 0.05) in C. jejuni counts were observed between samples at 10°C and those at 4°C. In contrast, the numbers of Salmonella Typhimurium in samples at 10°C had significantly (P < 0.05) increased (0.41 log CFU/g) from those at the refrigerated temperature of 4°C. Vacuum storage at 4 and 10°C for 28 days did not result in dramatic reductions in the mean numbers of C. jejuni and Salmonella Typhimurium. Our findings indicate that vacuum packaging under chilled conditions will not add substantially to safety for moisture-enhanced pork. Strict hygienic practices or the implementation of decontamination technologies is recommended.