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.     

Radiation processing to ensure safety of minimally processed carrot (Daucus carota) and cucumber (Cucumis sativus): optimization of dose for the elimination of Salmonella Typhimurium and Listeria monocytogenes

Minimally processed vegetables are in demand, because they offer convenience to consumers. However, these products are often unsafe because of possible contamination with pathogens, such as Salmonella, Escherichia coli O157:H7, and Shigella species. Therefore, this study was carried out to optimize the radiation dose necessary to ensure the safety of precut carrot and cucumber. Decimal reduction doses (D-values) of Salmonella Typhimurium MTCC 98 were ca. 0.164 kGy in carrot samples and 0.178 kGy in cucumber samples. D-values of Listeria monocytogenes were determined to be 0.312 and 0.345 kGy in carrot and cucumber samples, respectively. Studies of inoculated, packaged, minimally processed carrot and cucumber samples showed that treatment with a 1-kGy dose of gamma radiation eliminated up to 4 log CFU/g of Salmonella Typhimurium and 3 log CFU/g of L. monocytogenes. However, treatment with a 2-kGy dose was necessary to eliminate these pathogens by 5 log CFU/g. Storage studies showed that both Salmonella Typhimurium and L. monocytogenes were able to grow at 10 degrees C in inoculated control samples. Neither of these pathogens could be recovered from radiation-processed samples after storage for up to 8 days.     

Irradiation D-values for Escherichia coli O157:H7 and Salmonella sp. on inoculated broccoli seeds and effects of irradiation on broccoli sprout keeping quality and seed viability

Like alfalfa sprouts, broccoli sprouts can be a vehicle for bacterial pathogens, which can cause illness when they are consumed. The gamma irradiation process was used to reduce numbers of bacterial pathogens on broccoli sprouts and seeds, and the effect of this process on the seeds was studied. The irradiation destruct values for Salmonella sp. and for strains of Escherichia coli O157:H7 inoculated on broccoli seeds were determined. Results obtained in this study indicate that a dose of 2 kGy reduced total background counts for broccoli sprouts from 10(6) to 10(7) CFU/g to 10(4) to 10(5) CFU/g and increased the shelf life of the sprouts by 10 days. Yield ratio (wt/wt), germination percentage, sprout length, and thickness were measured to determine the effects of various irradiation doses on the broccoli seeds. Results show a decreased germination percentage at a dose level of 4 kGy, whereas the yield ratio (wt/wt), sprout length, and thickness decreased at the 2-kGy dose level. The radiation doses required to inactivate Salmonella sp. and strains of E. coli O157:H7 were higher than previously reported values. D-values, dose required for a 1-log reduction, for the nonvegetable and vegetable Salmonella sp. isolates were 0.74 and 1.10 kGy, respectively. The values for the nonvegetable and vegetable isolated strains of Escherichia coli O157:H7 were 1.43 and 1.11 kGy, respectively. With the irradiation process, a dose of up to 2 kGy can extend the shelf life of broccoli sprouts. A dose of > 2 kGy would have an adverse effect on the broccoli seed and decrease the yield of broccoli sprouts.     

Heat treatments to enhance the safety of mung bean seeds

Salmonella enterica serovars and Escherichia coli O157:H7 have been associated with contaminated seed sprout outbreaks. The majority of these outbreaks have been traced to sprout seeds contaminated with low levels of pathogens. E. coli O157:H7 strains can grow an average of 2.3 log CFU/g over 2 days during seed germination, and Salmonella can achieve an average growth of 3.7 log CFU/g. Therefore, it is important to find an effective method to reduce possible pathogenic bacterial populations on the seeds prior to sprouting. Our objective was to assess the effectiveness of various dry heat treatments on reducing E. coli O157:H7 and Salmonella populations on mung beans intended for sprout production and to determine the effect of these treatments on seed germination. Mung beans were inoculated with five-strain cocktails of E. coli O157:H7 and of Salmonella serovars harboring the green fluorescent protein gene and then air dried overnight. Heat treatments were performed by incubating the seeds at 55 degrees C for various periods of time. Heat-treated seeds were then assessed for the efficacy of the heat treatment and the effects of heat treatment on germination rates. After inoculation and drying, 6 log CFU/g E. coli O157:H7 and 4 log CFU/g Salmonella were detected on the seeds. Following heat treatment, pathogenic bacterial populations on the seeds were below detectable levels (<1 log CFU/g), but the germination rate of the seed was not affected. Thus, the risk of contamination and the presence of pathogens in the finished sprouts were greatly reduced via the seed heat treatment process.     

Effectiveness of radiation processing for elimination of Salmonella Typhimurium from minimally processed pineapple (Ananas comosus Merr.)

ABSTRACT:  The microbiological quality of market samples of minimally processed (MP) pineapple was examined. The effectiveness of radiation treatment in eliminating Salmonella Typhimurium from laboratory inoculated ready-to-eat pineapple slices was also studied. Microbiological quality of minimally processed pineapple samples from Mumbai market was poor; 8.8% of the samples were positive for Salmonella. D₁₀ (the radiation dose required to reduce bacterial population by 90%) value for S. Typhimurium inoculated in pineapple was 0.242 kGy. Inoculated pack studies in minimally processed pineapple showed that the treatment with a 2-kGy dose of gamma radiation could eliminate 5 log CFU/g of S. Typhimurium. The pathogen was not detected from radiation-processed samples up to 12 d during storage at 4 and 10 °C. The processing of market samples with 1 and 2 kGy was effective in improving the microbiological quality of these products.     

Shelf life extension of minimally processed cabbage and cucumber through gamma irradiation

The influence of irradiation of minimally processed cabbage and cucumber on microbial safety, texture, and sensory quality was investigated. Minimally processed, polyethylene-packed, and irradiated cabbage and cucumber were stored at refrigeration temperature (5 degrees C) for 2 weeks. The firmness values ranged from 3.23 kg (control) to 2.82 kg (3.0-kGy irradiated samples) for cucumbers, with a gradual decrease in firmness with increasing radiation dose (0 to 3 kGy). Cucumbers softened just after irradiation with a dose of 3.0 kGy and after 14 days storage, whereas the texture remained within acceptable limits up to a radiation dose of 2.5 kGy. The radiation treatment had no effect on the appearance scores of cabbage; however, scores decreased from 7.0 to 6.7 during storage. The appearance and flavor scores of cucumbers decreased with increasing radiation dose, and overall acceptability was better after radiation doses of 2.5 and 3.0 kGy. The aerobic plate counts per gram for cabbage increased from 3 to 5 log CFU (control), from 1.85 to 2.93 log CFU (2.5 kGy), and from a few colonies to 2.6 log CFU (3.0 kGy) after 14 days of storage at 5 degrees C. A similar trend was noted for cucumber samples. No coliform bacteria were detected at radiation doses greater than 2.0 kGy in either cabbage or cucumber samples. Total fungal counts per gram of sample were within acceptable limits for cucumbers irradiated at 3.0 kGy, and for cabbage no fungi were detected after 2.0-kGy irradiation. The D-values for Escherichia coli in cucumber and cabbage were 0.19 and 0.17 kGy, and those for Salmonella Paratyphi A were 0.25 and 0.29 kGy for cucumber and cabbage, respectively.     

Interaction of Salmonella enterica subspecies enterica serovar Typhimurium and mung bean (Phaseolus aureus) plants

The effect of Salmonella enterica subspecies enterica serovar Typhimurium, a zoonotic serovar, on mung bean (Phaseolus aureus) cultivar Pant Mung-3 plants was studied. Inoculation of mung bean seeds with Salmonella Typhimurium (7.2 x 10(5) CFU/ml) reduced sprouting rate (P < 0.07). This effect was more pronounced at higher levels of contamination. In the soil inoculated with Salmonella Typhimurium (7.2 x 10(6) CFU/g), germination was retarded and the number of defective sprouts was also significantly higher (P < 0.002). Salmonella Typhimurium grew inside germinating seeds and plant tissues and persisted in seedlings, adult plants, and harvested seedlings dried and stored at room temperature (30 degrees C) up to 45 days. Phaseolus aureus plants grown in sterile soil was resistant to Salmonella Typhimurium infection at 15 days of age and cleared Salmonella from all the aerial parts within 3 h of infection. However, Salmonella Typhimurium could be reisolated from the basal area of the stem and from soil even after 45 days of exposure to the pathogen.     

Hot water treatments to inactivate Escherichia coli O157:H7 and Salmonella in mung bean seeds

The majority of the seed sprout-related outbreaks have been associated with Escherichia coli O157:H7 and Salmonella. Therefore, an effective method is needed to inactivate these organisms on the seeds before they are sprouted. This study was conducted to assess the effectiveness of various hot water treatments to inactivate E. coli O157:H7 and Salmonella populations on mung beans seeds intended for sprout production and to determine the effect of these treatments on seed germination after the seeds were dipped in chilled water for 30 s. Mung bean seed inoculated with four-strain cocktails of E. coli O157:H7 and Salmonella were soaked into hot water at 80 and 90 degrees C with shaking for various periods and then dipped in chilled water for 30 s. The treated seeds were then assessed for the efficacy of the treatment for reducing populations of the pathogens and the effects of the treatment on germination. After inoculation and air drying, 6.08 +/- 0.34 log CFU/g E. coli O157:H7 and 5.34 +/- 0.29 log CFU/g Salmonella were detected on the seeds. After hot water treatment at 90 degrees C for 90 s followed by dipping in chilled water for 30 s, no viable pathogens were found and no survivors were found in the enrichment medium and during the sprouting process. The germination yield of the seed was not affected significantly. Therefore, hot water treatment followed by dipping in chilled water for 30 s could be an effective seed decontamination method for mung bean seeds intended for sprout production.     

Effect of freezing, irradiation, and frozen storage on survival of Salmonella in concentrated orange juice

Six strains of Salmonella (Anatum F4317, Dublin 15480, Enteritidis 13076, Enteritidis WY15159, Stanley H0588, and Typhimurium 14028) were individually inoculated into orange juice concentrate (OJC) and frozen to -20 degrees C. The frozen samples were treated with 0 (nonirradiated), 0.5, 1.0, or 2.0 kGy of gamma radiation and held frozen for 1 h, and the surviving bacterial population was assessed. The strains showed significant variability in their response to freezing and to freezing in combination with irradiation. The response was dose dependent. Relative to the nonfrozen, nonirradiated control, the reduction following the highest dose (2.0 kGy) ranged from 1.29 log CFU/ml (Salmonella Typhimurium) to 2.17 log CFU/ml (Salmonella Stanley). Samples of OJC inoculated with Salmonella Enteritidis WY15159 and irradiated were stored at -20 degrees C for 1, 2, 7, or 14 days, and the surviving population was determined. Relative to the nonfrozen, nonirradiated control, after 14 days, the population was reduced by 1.2 log CFU/ml in the nonirradiated samples and by 3.3 log CFU/ml following treatment with 2.0 kGy. The combination of frozen storage plus irradiation resulted in greater overall reductions than either process alone.     

Inactivation of Escherichia coli O157:H7 and Salmonella typhimurium DT 104 on alfalfa seeds by levulinic acid and sodium dodecyl sulfate

Studies were conducted to determine the best concentration and exposure time for treatment of alfalfa seeds with levulinic acid plus sodium dodecyl sulfate (SDS) to inactivate Escherichia coli O157:H7 and Salmonella without adversely affecting seed germination. Alfalfa seeds inoculated with a five-strain mixture of E. coli O157:H7 or Salmonella Typhimurium were dried in a laminar flow hood at 21°C for up to 72 h. Inoculated alfalfa seeds dried for 4 h then treated for 5 min at 21°C with 0.5% levulinic acid and 0.05% SDS reduced the population of E. coli O157:H7 and Salmonella Typhimurium by 5.6 and 6.4 log CFU/g, respectively. On seeds dried for 72 h, treatment with 0.5% levulinic acid and 0.05% SDS for 20 min at 21°C reduced E. coli O157:H7 and Salmonella Typhimurium populations by 4 log CFU/g. Germination rates of alfalfa seeds treated with 0.5% levulinic acid plus 0.05% SDS for up to 1 h at 21°C were compared with a treatment of 20,000 ppm of calcium hypochlorite or tap water only. Treatment of alfalfa seeds with 0.5% levulinic acid plus 0.05% SDS for 5 min at 21°C resulted in a >3.0-log inactivation of E. coli O157:H7 and Salmonella.     

A rapid and automated fiber optic-based biosensor assay for the detection of Salmonella in spent irrigation water used in the sprouting of sprout seeds

Recent outbreaks of foodborne illness have been linked to the consumption of contaminated sprouts. The spent irrigation water used to irrigate sprouts can carry many microorganisms, including pathogenic strains of Escherichia coli and Salmonella enterica. These pathogens are believed to originate from the seeds. The U.S. Food and Drug Administration recommends that sprout producers conduct microbiological testing of spent irrigation water from each production lot at least 48 h after seeds have germinated. Microbial analysis for the detection of Salmonella is labor-intensive and takes days to complete. A rapid and automated fiber-optic biosensor assay for the detection of Salmonella in sprout rinse water was developed in this study. Alfalfa seeds contaminated with various concentrations of Salmonella Typhimurium were sprouted. The spent irrigation water was assayed 67 h after alfalfa seed germination with the RAPTOR (Research International, Monroe, Wash.), an automated fiber optic-based detector. Salmonella Typhimurium could be positively identified in spent irrigation water when seeds were contaminated with 50 CFU/g. Viable Salmonella Typhimurium cells were also recovered from the waveguides after the assay. This biosensor assay system has the potential to be directly connected to water lines within the sprout-processing facility and to operate automatically, requiring manual labor only for preventative maintenance. Therefore, the presence of Salmonella Typhimurium in spent irrigation water could be continuously and rapidly detected 3 to 5 days before the completion of the sprouting process.     

Use of green fluorescent protein expressing Salmonella Stanley to investigate survival, spatial location, and control on alfalfa sprouts.

Laser scanning confocal microscopy (LSCM) was used to observe the interaction of Salmonella Stanley with alfalfa sprouts. The green fluorescent protein (gfp) gene was integrated into the chromosome of Salmonella Stanley for constitutive expression, thereby eliminating problems of plasmid stability and loss of signal. Alfalfa seeds were inoculated by immersion in a suspension of Salmonella Stanley (ca. 10(7) CFU/ml) for 5 min at 22 degrees C. Epifluorescence microscopy demonstrated the presence of target bacteria on the surface of sprouts. LSCM demonstrated bacteria present at a depth of 12 microm within intact sprout tissue. An initial population of ca. 10(4) CFU/g seed increased to 7.0 log CFU/g during a 24-h germination period and then decreased to 4.9 log CFU/g during a 144-h sprouting period. Populations of Salmonella Stanley on alfalfa seeds decreased from 5.2 to 4.1 log CFU/g and from 5.2 to 2.8 log CFU/g for seeds stored 60 days at 5 and 22 degrees C, respectively. The efficacy of 100, 200, 500, or 2,000 ppm chlorine in killing Salmonella Stanley associated with sprouts was determined. Treatment of sprouts in 2,000 ppm chlorine for 2 or 5 min caused a significant reduction in populations of Salmonella Stanley. Influence of storage on Salmonella Stanley populations was investigated by storing sprouts 4 days at 4 degrees C. The initial population (7.76 log CFU/g) of Salmonella Stanley on mature sprouts decreased (7.67 log CFU/g) only slightly. Cross-contamination during harvest was investigated by harvesting contaminated sprouts, then directly harvesting noncontaminated sprouts. This process resulted in the transfer of ca. 10(5) CFU/g Salmonella Stanley to the noncontaminated 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.     

Alfalfa seed germination and yield ratio and alfalfa sprout microbial keeping quality following irradiation of seeds and sprouts.

Foods can be treated with gamma radiation, a nonthermal food process, to inactivate foodborne pathogens and fungi, to kill insects on or in fruits and vegetables, and to increase shelf life. Gamma irradiation is especially well suited for these treatments because of its ability to penetrate commercial pallets of foods. Irradiated fruits, vegetables, poultry, and hamburger have been received favorably by the public and are now available in supermarkets. The use of irradiation on fresh alfalfa sprouts was studied to determine its effect on keeping quality as related to aerobic microbial load. After an irradiation dose of 2 kGy, the total aerobic count decreased from 10(5-8) to 10(3-5) CFU/g, and the total coliform counts decreased from 10(5-8) to 10(3-0) CFU/g. The results showed that the sprouts maintained their structure after irradiation, and the keeping quality was extended to 21 days, which is an increase of 10 days from the usual shelf life. The effect of various doses of irradiation on alfalfa seeds as measured by percent germination and yield ratio (wt/wt) of sprouts was determined. There was little effect on the percent germination, but as the dose increased, the yield ratio of alfalfa sprouts decreased. As the length of growing time increased, so did the yield ratio of the lower dose irradiated seeds (1 to 2 kGy). The irradiation process can be used to increase the shelf life of alfalfa sprouts, and irradiating alfalfa seeds at doses up to 2 kGy does not unacceptably decrease the yield ratio for production of alfalfa sprouts.     

Thermal inactivation of Salmonella and Escherichia coli O157:H7 on alfalfa seeds

Alfalfa seeds inoculated with five strains of Salmonella or Escherichia coli O157:H7 were subjected to dry heat at 55 degrees C for up to 8 days. Five-log reductions in Salmonella or E. coli O157:H7 on seeds were observed. No pathogens were detected on the sprouted seeds, which were initially inoculated with ca. 2 log CFU/g of Salmonella or more than 8 log CFU/g of E. coli O157:H7. The percentages of germination of the alfalfa seeds did not significantly decrease after 6 days of heating at 55 degrees C. These results showed that heat treatment of alfalfa seeds at 55 degrees C for up to 6 days was effective in enhancing the safety of alfalfa sprouts without affecting germination significantly.     

Microbial Quality and Safety of X-Ray Irradiated Fresh Catfish (Ictalurus punctatus) Fillets Stored under CO(2) Atmosphere

The effect of low-dose X-ray irradiation (0, 2, and 3 kGy) on pathogens, spoilage bacteria (psychrotrophic, anaerobic, and lactic acid bacteria), and physicochemical parameters of iced catfish fillets stored under carbon dioxide (CO(2) ) atmosphere were studied. Irradiation of 2 or 3 kGy eliminated all Listeria monocytogenes (4.8 log CFU/g) and Salmonella Typhimurium (4.7 log CFU/g) while CO(2) atmosphere alone showed less than 1 log reduction on Salmonella Typhimurium. There was no recovery of either pathogen for the experimental period. Regardless of dose, irradiation showed 2 log reduction on psychrotrophic, anaerobic, and lactic acid bacteria for 24 d. Further post-irradiation reductions of spoilage bacteria were observed. Spoilage bacteria on catfish fillets treated with 2 kGy irradiation regained their growth after 16 d of storage. However, catfish fillets exposed to 3 kGy did not show growth of spoilage bacteria throughout the experimental period (24 d). Irradiation (2 or 3 kGy) increased pH and TBARS, and decreased Hunter "b" values but produced no effect on water holding capacity and texture. Shelf life of catfish fillets exposed to 2 or 3 kGy irradiation could be extended by more than 24 d at 2 to 4 °C, with the added safety of possibly eliminating pathogens.     

Assessment of the potential for Listeria monocytogenes survival and growth during alfalfa sprout production and use of ionizing radiation as a potential intervention treatment

Alfalfa seeds (Australian, nondormant, nonscarified) were treated with 20,000 ppm active chlorine, sprouted in canning jars for 5 days, and packaged and stored at 5 degrees C for up to 9 days. Seeds or sprouts were inoculated with a three-strain cocktail of Listeria monocytogenes at one of three points during the process-day 0 (before 24-h aqueous seed soak), day 1 (after 24-h aqueous seed soak), or day 5 (after sprouting, before prepackaging 10 ppm chlorine rinse)--or control (no inoculum), and the ability of the inoculum to survive and grow was evaluated. Total bacterial numbers on uninoculated seeds increased dramatically during the first 24-h the seeds were soaked, from 3.5 to ca. 8.0 log CFU/g, and remained at this level during refrigerated storage. When the seeds were inoculated with a cocktail of L. monocytogenes (log 5 CFU/10 ml) on day 0 or 1, the population of the pathogen increased dramatically, to within 1 to 2 logs of the total, and remained high during refrigerated storage. When sprouted seeds were inoculated with L. monocytogenes later in the process (day 5), the inoculum survived but did not grow more than ca. 1 log CFU/g, regardless of whether the inoculation level in each jar was low (10(3)) or high (10(5)). Irradiation of sprouts with beta radiation at 3.3 or 5.3 kGy, but not 1.5 kGy, was effective at eliminating L. monocytogenes from inoculated sprouts (6 log CFU/g) without causing noticeable changes in appearance or odor. In summary, L. monocytogenes can grow on sprouts during production, can survive on refrigerated sprouts, and may be eliminated on sprouts with beta radiation.     

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.     

Chemical and irradiation treatments for killing Escherichia coli O157:H7 on alfalfa,radish, and mung bean seeds

In this study, the effectiveness of dry-heat treatment in combination with chemical treatments (electrolyzed oxidizing [EO] water, califresh-S, 200 ppm of active chlorinated water) with and without sonication in eliminating Escherichia coli O157:H7 on laboratory-inoculated alfalfa, radish, and mung bean seeds was compared with that of dry-heat treatment in combination with irradiation treatment. The treatment of mung bean seeds with EO water in combination with sonication followed by a rinse with sterile distilled water resulted in reductions of approximately 4.0 log10 CFU of E. coli O157:H7 per g. whereas reductions of ca. 1.52 and 2.64 log10 CFU/g were obtained for radish and alfalfa seeds. The maximum reduction (3.70 log10 CFU/g) for mung bean seeds was achieved by treatment with califresh-S and chlorinated water (200 ppm) in combination with sonication and a rinse. The combination of dry heat, hot EO water treatment, and sonication was able to eliminate pathogen populations on mung bean seeds but was unable to eliminate the pathogen on radish and alfalfa seeds. Other chemical treatments used were effective in greatly reducing pathogen populations on radish and alfalfa seeds without compromising the quality of the sprouts, but these treatments did not result in the elimination of pathogens from radish and alfalfa seeds. Moreover, a combination of dry-heat and irradiation treatments was effective in eliminating E. coli O157:H7 on laboratory-inoculated alfalfa, radish, and mung bean seeds. An irradiation dose of 2.0 kGy in combination with dry heat eliminated E. coli O157:H7 completely from alfalfa and mung bean seeds, whereas a 2.5-kGy dose of irradiation was required to eliminate the pathogen completely from radish seeds. Dry heat in combination with irradiation doses of up to 2.0 kGy did not unacceptably decrease the germination percentage for alfalfa seeds or the length of alfalfa sprouts but did decrease the lengths of radish and mung bean sprouts.     

Evaluation of volatile chemical treatments for lethality to Salmonella on alfalfa seeds and sprouts

A study was done to evaluate natural volatile compounds for their ability to kill Salmonella on alfalfa seeds and sprouts. Acetic acid, allyl isothiocyanate (AIT), trans-anethole, carvacrol, cinnamic aldehyde, eugenol, linalool, methyl jasmonate, and thymol were examined for inhibitory and lethal activity against Salmonella by exposing inoculated alfalfa seeds to compounds (1,000 mg/liter of air) for 1, 3, and 7 h at 60 degrees C. Only acetic acid, cinnamic aldehyde, and thymol caused significant reductions in Salmonella populations (>3 log10 CFU/g) compared with the control (1.9 log10 CFU/g) after treatment for 7 h. Treatment of seeds at 50 degrees C for 12 h with acetic acid (100 and 300 mg/liter of air) and thymol or cinnamic aldehyde (600 mg/liter of air) significantly reduced Salmonella populations on seeds (>1.7 log10 CFU/g) without affecting germination percentage. Treatment of seeds at 50 degrees C with AIT (100 and 300 mg/liter of air) and cinnamic aldehyde or thymol (200 mg/liter of air) did not significantly reduce populations compared with the control. Seed germination percentage was largely unaffected by treatment with gaseous acetic acid, AIT, cinnamic aldehyde, or thymol for up to 12 h at 50 degrees C. The number of Salmonella on seeds treated at 70 degrees C for 80 min with acetic acid (100 and 300 mg/liter of air), AIT (100 mg/liter of air), and cinnamic aldehyde and thymol (600 mg/liter of air) at water activity (a(w)) 0.66 was not significantly different than the number inactivated on seeds at a(w) 0.49. Acetic acid at 200 and 500 mg/liter of air reduced an initial population of 7.50 log10 CFU/g of alfalfa sprouts by 2.33 and 5.72 log10 CFU/g, respectively, within 4 days at 10 degrees C. whereas AIT at 200 and 500 mg/liter of air reduced populations to undetectable levels; however, both treatments caused deterioration in sensory quality. Treatment of sprouts with 1 or 2 mg of AIT per liter of air adversely affected sensory quality but did not reduce Salmonella populations after 11 days of exposure at 10 degrees C.