Robert E. Brackett

Incidence, contributing factors, and control of bacterial pathogens in produce

Abstract The importance of bacterial pathogens in the transmission of foodborne illness has become apparent in recent years. Several large, well-publicized outbreaks of foodborne illness have been linked to cantaloupe, tomatoes, lettuce, alfalfa sprouts, and both apple and orange juices. In addition, numerous other smaller scale outbreaks linked to these and other commodities have also been reported. Although contributing factors have not been determined in all cases, several notable causes have been proposed. In particular, cross contamination with fecal matter of both domestic as well as wild animals have been suggested. In addition, contact with contaminated water has also been identified as a source of contamination. However, the use of untreated manure or sewage, lack of field sanitation, poorly or unsanitized transportation vehicles, and contamination by handlers are also suggested as potential contributing factors. Control of foodborne pathogens in produce must begin before produce is even planted by avoiding fields which have been subjected to flooding, on which animals have been recently grazed, or have otherwise been contaminated with manure. After planting, only clean potable water should be used for irrigation and harvesting equipment should be thoroughly cleaned and sanitized. Both field workers and packinghouse and processing plant personnel should be instructed in proper personal hygiene and provided with adequate sanitary and handwashing facilities. Vehicles transporting finished products should be sanitized, properly loaded to provide adequate air circulation, and maintained at proper temperatures. Likewise, retail display cases must be kept clean and at proper refrigeration temperatures. Finally, consumers should be informed as to proper handling of produce, particularly in the case of new generation products such as modified atmosphere packaged produce.

Roles of Oxidation-Reduction Potential in Electrolyzed Oxidizing and Chemically Modified Water for the Inactivation of Food-Related Pathogens

This study investigates the properties of electrolyzed oxidizing (EO) water for the inactivation of pathogen and to evaluate the chemically modified solutions possessing properties similar to EO water in killing Escherichia coli O157:H7. A five-strain cocktail (10(10) CFU/ml) of E. coli O157:H7 was subjected to deionized water (control), EO water with 10 mg/liter residual chlorine (J.A.W-EO water), EO water with 56 mg/liter residual chlorine (ROX-EO water), and chemically modified solutions. Inactivation (8.88 log10 CFU/ml reduction) of E. coli O157:H7 occurred within 30 s after application of EO water and chemically modified solutions containing chlorine and 1% bromine. Iron was added to EO or chemically modified solutions to reduce oxidation-reduction potential (ORP) readings and neutralizing buffer was added to neutralize chlorine. J.A.W-EO water with 100 mg/liter iron, acetic acid solution, and chemically modified solutions containing neutralizing buffer or 100 mg/liter iron were ineffective in reducing the bacteria population. ROX-EO water with 100 mg/liter iron was the only solution still effective in inactivation of E. coli O157:H7 and having high ORP readings regardless of residual chlorine. These results suggest that it is possible to simulate EO water by chemically modifying deionized water and ORP of the solution may be the primary factor affecting microbial inactivation.

Shelf stability and safety of fresh produce as influenced by sanitation and disinfection.

Quality and safety of fresh produce depend on their microbiological flora. Every step from production through consumption will influence the microbiology of fresh produce. Improper handling and unsanitary equipment lead to increased populations of microorganisms on fresh fruits and vegetables and can compromise quality and safety. Processing steps such as cutting and peeling usually increase the population of microorganisms and shorten shelf life. Using techniques to extend shelf life can increase the risk of safety problems developing and therefore need to be carefully evaluated. Proper use of disinfectants can complement an effective sanitation program but should not be relied upon to eliminate pathogenic microorganisms from contaminated produce.

Efficacy of Electrolyzed Oxidizing (EO) and Chemically Modified Water on Different Types of Foodborne Pathogens

This study was undertaken to evaluate the efficacy of electrolyzed oxidizing (EO) and chemically modified water with properties similar to the EO water for inactivation of different types of foodborne pathogens (Escherichia coli O157:H7, Listeria monocytogenes and Bacillus cereus). A five-strain cocktail of each microorganism was exposed to deionized water (control), EO water and chemically modified water. To evaluate the effect of individual properties (pH, oxidation-reduction potential (ORP) and residual chlorine) of treatment solutions on microbial inactivation, iron was added to reduce ORP readings and neutralizing buffer was added to neutralize chlorine. Inactivation of E. coli O157:H7 occurred within 30 s after application of JAW EO water with 10 mg/l residual chlorine and chemically modified solutions containing 13 mg/l residual chlorine. Inactivation of Gram-positive and -negative microorganisms occurred within 10 s after application of ROX EO water with 56 mg/l residual chlorine and chemically modified solutions containing 60 mg/l residual chlorine. B. cereus was more resistant to the treatments than E. coli O157:H7 and L. monocytogenes and only 3 log10 reductions were achieved after 10 s of ROX EO water treatment. B. cereus spores were the most resistant pathogen. However, more than 3 log10 reductions were achieved with 120-s EO water treatment.

Survival of Salmonellae on and in Tomato Plants from the Time of Inoculation at Flowering and Early Stages of Fruit Development through Fruit Ripening

ABSTRACT The fate of salmonellae applied to tomato plants was investigated. Five Salmonella serotypes were used to inoculate tomato plants before and after fruits set, either by injecting stems with inoculum or brushing flowers with it. Ripe tomato fruits were subjected to microbiological analysis. Peptone wash water, homogenates of stem scar tissues, and homogenates of fruit pulp were serially diluted and plated on bismuth sulfite agar before and after enrichment. Presumptive Salmonella colonies were confirmed by serological tests, PCR assay using HILA2 primers, and enterobacterial repetitive intergenic consensus PCR. Of 30 tomatoes harvested from inoculated plants, 11 (37%) were positive forSalmonella. Of the Salmonella-positive tomatoes, 43 and 40%, respectively, were from plants receiving stem inoculation before and after flower set. Two of eight tomatoes produced from inoculated flowers contained Salmonella. Higher percentages of surface (82%) and stem scar tissue (73%) samples, compared to pulp of Salmonella-positive tomatoes (55%), harbored the pathogen. Of the five serotypes in the inoculum, Montevideo was the most persistent, being isolated from tomatoes 49 days after inoculation, and Poona was the most dominant, being present in 5 of 11 Salmonella-positive tomatoes. Results suggest that Salmonella cells survive in or on tomato fruits from the time of inoculation at flowering through fruit ripening. Tomato stems and flowers are possible sites at whichSalmonella may attach and remain viable during fruit development, thus serving as routes or reservoirs for contaminating ripened fruit.

Evidence of Association of Salmonellae with Tomato Plants Grown Hydroponically in Inoculated Nutrient Solution

ABSTRACT The possibility of uptake of salmonellae by roots of hydroponically grown tomato plants was investigated. Within 1 day of exposure of plant roots to Hoagland nutrient solution containing 4.46 to 4.65 log10 CFU of salmonellae/ml, the sizes of the pathogen populations were 3.01 CFU/g of hypocotyls and cotyledons and 3.40 log10 CFU/g of stems for plants with intact root systems (control) and 2.55 log10 CFU/g of hypocotyls and cotyledons for plants from which portions of the roots had been removed. A population of ≥3.38 log10 CFU/g of hypocotyls-cotyledons, stems, and leaves of plants grown for 9 days was detected regardless of the root condition. Additional studies need to be done to unequivocally demonstrate that salmonellae can exist as endophytes in tomato plants grown under conditions that simulate commonly used agronomic practices.

Growth of Listeria monocytogenes on fresh vegetables stored under controlled atmosphere

The effects of controlled atmosphere storage (CAS) on survival and growth of Listeria monocytogenes on fresh asparagus, broccoli, and cauliflower were investigated. Vegetables were inoculated with L. monocytogenes strain Scott A or LCDC 81-861 (103-105 CFU/g) and stored at 4 and 15°C under CAS and air. L. monocytogenes populations were monitored over 21 day (4°C) and 10 day (15°C) periods using selective recovery media and a direct plating technique. CAS lengthened the time that all vegetables were considered acceptable for consumption by subjective inspection. Populations of L. monocytogenes increased during storage but CAS did not influence the rate of growth.

Efficacy of Electrolyzed Oxidizing Water in Inactivating Salmonella on Alfalfa Seeds and Sprouts

Studies have demonstrated that electrolyzed oxidizing (EO) water is effective in reducing foodborne pathogens on fresh produce. This study was undertaken to determine the efficacy of EO water and two different forms of chlorinated water (chlorine water from Cl2 and Ca(OCl)2 as sources of chlorine) in inactivating Salmonella on alfalfa seeds and sprouts. Tengram sets of alfalfa seeds inoculated with a five-strain cocktail of Salmonella (6.3 x 10(4) CFU/g) were subjected to 90 ml of deionized water (control), EO water (84 mg/liter of active chlorine), chlorine water (84 mg/liter of active chlorine), and Ca(OCl)2 solutions at 90 and 20,000 mg/liter of active chlorine for 10 min at 24 +/- 2 degrees C. The application of EO water, chlorinated water, and 90 mg/liter of Ca(OCl)2 to alfalfa seeds for 10 min reduced initial populations of Salmonella by at least 1.5 log10 CFU/g. For seed sprouting, alfalfa seeds were soaked in the different treatment solutions described above for 3 h. Ca(OCl)2 (20,000 mg/liter of active chlorine) was the most effective treatment in reducing the populations of Salmonella and non-Salmonella microflora (4.6 and 7.0 log10 CFU/g, respectively). However, the use of high concentrations of chlorine generates worker safety concerns. Also, the Ca(OCl)2 treatment significantly reduced seed germination rates (70% versus 90 to 96%). For alfalfa sprouts, higher bacterial populations were recovered from treated sprouts containing seed coats than from sprouts with seed coats removed. The effectiveness of EO water improved when soaking treatments were applied to sprouts in conjunction with sonication and seed coat removal. The combined treatment achieved 2.3- and 1.5-log10 CFU/g greater reductions than EO water alone in populations of Salmonella and non-Salmonella microflora, respectively. This combination treatment resulted in a 3.3-log10 CFU/g greater reduction in Salmonella populations than the control (deionized water) treatment.

Physical, Chemical and Biological Degradation of Mycotoxins in Foods and Agricultural Commodities

Aflatoxin is partially or completely degraded by irradiation, heat, or treatment with strong acids or bases, oxidizing agents or bisulfite. Hydrogen peroxide plus riboflavin denature aflatoxin in milk. Mycelia of Aspergillus parasiticus can degrade aflatoxin, possibly via fungal peroxidase. Such degradation is affected by strain of A. parasiticus , amount of mycelium, temperature, pH and concentration of aflatoxin. Adsorbants, including bentonite and activated charcoal, can physically remove aflatoxin and patulin from liquid foods. Patulin is stable at low pH values but not in the presence of large amounts of vitamin C or bisulfite. Patulin can be degraded by actively fermenting yeasts and rubratoxin can be degraded by the mycelium of Penicillium rubrum .

Survival of Salmonella on Tomatoes Stored at High Relative Humidity, in Soil, and on Tomatoes in Contact with Soil

Salmonellosis has been linked to the consumption of several types of raw fruits and vegetables, some of which may have been contaminated with Salmonella before harvesting. The objectives of this study were to investigate water and soil as reservoirs of Salmonella for the contamination of mature green tomato fruits. Salmonella survived for at least 45 days in inoculated moist soil. The population of Salmonella on tomatoes in contact with soil increased by 2.5 log10 CFU per tomato during storage for 4 days at 20 degrees C and remained constant for an additional 10 days. The number of cells inoculated on tomatoes decreased by approximately 4 log10 CFU per tomato during storage for 14 days at 20 degrees C and 70% relative humidity. Fruits in contact with inoculated soil for 1 day at 20 degrees C harbored Salmonella only near or on the skin surface. More Salmonella cells were observed in stem scar and subsurface areas of tomatoes as the time of storage increased. PCR fingerprinting revealed that among five Salmonella serotypes in the inoculum, Salmonella Montevideo was the most persistent on tomatoes in contact with inoculated soil and on spot-inoculated tomatoes, followed by Salmonella Poona and Salmonella Michigan. The results of this study demonstrate that an enhanced green fluorescent protein marker can be used to detect cells and monitor the growth of Salmonella in the presence of other microorganisms. Observations on the infiltration of Salmonella into tomato tissues support the contention that preharvest contact of produce with contaminated water or soil exacerbates problems associated with the postharvest removal of pathogens or their accessibility to treatment with sanitizers.