Survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on Inoculated Almonds and Pistachios Stored at -19, 4, and 24° C

The survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes was determined on almonds and pistachios held at typical storage temperatures. Almond kernels and inshell pistachios were inoculated with four- to six-strain cocktails of nalidixic acid-resistant Salmonella, E. coli O157:H7, or L. monocytogenes at 6 log CFU/g and then dried for 72 h. After drying, inoculated nuts were stored at -19, 4, or 24°C for up to 12 months. During the initial drying period after inoculation, levels of all pathogens declined by 1 to -log CFU/g on both almonds and pistachios. During storage, moisture content (4.8%) and water activity (0.4) of the almonds and pistachios were consistent at -19°C; increased slowly to 6% and 0.6, respectively, at 4°C; and fluctuated from 4 to 5% and 0.3 to 0.5 at 24°C, respectively. Every 1 or 2 months, levels of each pathogen were enumerated by plating; samples were enriched when levels fell below the limit of detection. No reduction in population level was observed at -19 or 4°C for either pathogen, with the exception of E. coli O157:H7-inoculated almonds stored at 4°C (decline of 0.09 log CFU/g/month). At 24°C, initial rates of decline were 0.20, 0.60, and 0.71 log CFU/g/month on almonds and 0.15, 0.35, and 0.86 log CFU/g/month on pistachios for Salmonella, E. coli O157:H7, and L. monocytogenes, respectively, but distinct tailing of the survival curves was noted for both E. coli O157:H7 and L. monocytogenes.     

Survival of Salmonella enteritidis phage type 30 on inoculated almonds stored at -20, 4, 23, and 35 degrees C

To evaluate the 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 degrees 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 degrees C, 4 +/- 2 degrees C, 23 +/- 3 degrees C, and 35 +/- 2 degrees C. No significant reductions of Salmonella were observed during storage at -20 and 4 degrees C over 550 days. At 35 degrees 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 degrees C, reductions of 0.18 and 0.30 log CFU/month were calculated for 171 and 550 days of storage, respectively. When combined with data from the study of inoculum levels, an overall average calculated reduction at 23 degrees 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 degrees 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 commonstorage conditions.     

A dry-inoculation method for nut kernels

A dry-inoculation method for almonds and walnuts was developed to eliminate the need for the postinoculation drying required for wet-inoculation methods. The survival of Salmonella enterica Enteritidis PT 30 on wet- and dry-inoculated almond and walnut kernels stored under ambient conditions (average: 23 °C; 41 or 47% RH) was then compared over 14 weeks. For wet inoculation, an aqueous Salmonella preparation was added directly to almond or walnut kernels, which were then dried under ambient conditions (3 or 7 days, respectively) to initial nut moisture levels. For the dry inoculation, liquid inoculum was mixed with sterilized sand and dried for 24 h at 40 °C. The dried inoculated sand was mixed with kernels, and the sand was removed by shaking the mixture in a sterile sieve. Mixing procedures to optimize the bacterial transfer from sand to kernel were evaluated; in general, similar levels were achieved on walnuts (4.8–5.2 log CFU/g) and almonds (4.2–5.1 log CFU/g). The decline of Salmonella Enteritidis populations was similar during ambient storage (98 days) for both wet-and dry-inoculation methods for both almonds and walnuts. The dry-inoculation method mimics some of the suspected routes of contamination for tree nuts and may be appropriate for some postharvest challenge studies.     

Inactivation of Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes on apples, oranges, and tomatoes by lactic acid with hydrogen peroxide.

The objective of this study was to develop a practical and effective method for inactivating or substantially reducing Escherichia coli O157:H7, Salmonella Enteritidis, and Listeria monocytogenes on apples, oranges, and tomatoes. Apples, oranges, and tomatoes were spot-inoculated with five-strain mixtures of E. coli O157:H7, Salmonella Enteritidis, and L. monocytogenes near the stem end and were submerged in sterile deionized water containing 1.5% lactic acid plus 1.5% hydrogen peroxide for 15 min at 40 degrees C. Inoculated samples treated with sterile deionized water at the same temperature and for the same duration served as controls. The bacterial pathogens on fruits subjected to the chemical treatment were reduced by >5.0 log10 CFU per fruit, whereas washing in deionized water decreased the pathogens by only 1.5 to 2.0 log10 CFU per fruit. Furthermore, substantial populations of the pathogens survived in the control wash water, whereas no E. coli O157:H7, Salmonella Enteritidis, or L. monocytogenes cells were detected in the chemical treatment solution. The sensory and qualitative characteristics of apples treated with the chemical wash solution were not adversely affected by the treatment. It was found that the treatment developed in this study could effectively be used to kill E. coli O157:H7, Salmonella Enteritidis, and L. monocytogenes on apples, oranges, and tomatoes at the processing or packaging level.     

Evaluation of gaseous chlorine dioxide as a sanitizer for killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, and yeasts and molds on fresh and fresh-cut produce

Gaseous chlorine dioxide (ClO2) was evaluated for effectiveness in killing Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on fresh-cut lettuce, cabbage, and carrot and Salmonella, yeasts, and molds on apples, peaches. tomatoes, and onions. Inoculum (100 microl, ca. 6.8 log CFU) containing five serotypes of Salmonella enterica, five strains of E. coli O157:H7, or five strains of L. monocytogenes was deposited on the skin and cut surfaces of fresh-cut vegetables, dried for 30 min at 22 degrees C, held for 20 h at 4 degrees C, and then incubated for 30 min at 22 degrees C before treatment. The skin surfaces of apples, peaches, tomatoes, and onions were inoculated with 100 microl of a cell suspension (ca. 8.0 log CFU) containing five serotypes of Salmonella, and inoculated produce was allowed to dry for 20 to 22 h at 22 degrees C before treatment. Treatment with ClO2 at 4.1 mg/liter significantly (alpha = 0.05) reduced the population of foodborne pathogens on all produce. Reductions resulting from this treatment were 3.13 to 4.42 log CFU/g for fresh-cut cabbage, 5.15 to 5.88 log CFU/g for fresh-cut carrots, 1.53 to 1.58 log CFU/g for fresh-cut lettuce, 4.21 log CFU per apple, 4.33 log CFU per tomato, 1.94 log CFU per onion, and 3.23 log CFU per peach. The highest reductions in yeast and mold populations resulting from the same treatment were 1.68 log CFU per apple and 2.65 log CFU per peach. Populations of yeasts and molds on tomatoes and onions were not significantly reduced by treatment with 4.1 mg/liter ClO2. Substantial reductions in populations of pathogens on apples, tomatoes, and onions but not peaches or fresh-cut cabbage, carrot, and lettuce were achieved by treatment with gaseous ClO2 without markedly adverse effects on sensory qualities.     

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.     

Antibacterial effect of water-soluble tea extracts on foodborne pathogens in laboratory medium and in a food model

The microbial inhibition of foodborne pathogens was determined in brain heart infusion broth with 10% (wt/vol) water-soluble extracts of green, jasmine, black, dungglre, and oolong tea against Escherichia coli O157:H7, Salmonella enterica serovar Enteritidis, Listeria monocytogenes, and Staphylococcus aureus. The mixed culture (approximately 6.0 log CFU/ml), which was composed of the four pathogens, was inoculated into brain heart infusion broth with and without tea extracts. After incubation at 35 degrees C for 0, 1, 3, and 5 days, proper dilution of each sample was spiral plated on each selective agar. Viable cell counts were performed after incubation at 35 degrees C for 24 to 36 h. Green, jasmine, and black tea exhibited an approximately 5.0 log suppression of S. aureus compared with the control from days 1 to 5. Green and jasmine tea also suppressed the growth of L. monocytogenes by approximately 3.0 log CFU/ml on day 5. In contrast, no tea extracts inactivated E. coli O157:H7 and Salmonella Enteritidis. Based on the result in liquid medium, green and jasmine teas of 0.1% (vol/wt) were individually evaluated for their antimicrobial activity against L. monocytogenes and S. aureus in a food model (ground beef) stored at 7 degrees C for 0, 1, 3, 5, and 7 days. Viable cell counts of total bacteria, L. monocytogenes, and S. aureus in ground beef were not significantly different among green and jasmine tea and the control.     

Viability of Salmonella,Escherichia coli O157:H7, and Listeria monocytogenes in yellow fat spreads as affected by storage temperature

A study was conducted to characterize the survival and inactivation kinetics of a five-serotype mixture of Salmonella (6.23 to 6.55 log10 CFU per 3.5-ml or 4-g sample), a five-strain mixture of Escherichia coli O157:H7 (5.36 to 6.14 log10 CFU per 3.5-ml or 4-g sample), and a six-strain mixture of Listeria monocytogenes (5.91 to 6.18 log10 CFU per 3.5-ml or 4-g sample) inoculated into seven yellow fat spreads (one margarine, one butter-margarine blend, and five dairy and nondairy spreads and toppings) after formulation and processing and stored at 4.4, 10, and 21 degrees C for up to 94 days. Neither Salmonella nor E. coil O157:H7 grew in any of the test products. The time required for the elimination of each pathogen depended on the product and the storage temperature. Death was more rapid at 21 degrees C than at 4.4 or 10 degrees C. Depending on the product, the time required for the elimination of viable cells at 21 degrees C ranged from 5 to 7 days to >94 days for Salmonella, from 3 to 5 days to 28 to 42 days for E. coli O157:H7, and from 10 to 14 days to >94 days for L. monocytogenes. Death was most rapid in a water-continuous spray product (pH 3.66, 4.12% salt) and least rapid in a butter-margarine blend (pH 6.66, 1.88% salt). E. coli O157:H7 died more rapidly than did Salmonella or L. monocytogenes regardless of storage temperature. Salmonella survived longer in high-fat (> or = 61%) products than in products with lower fat contents. The inhibition of growth is attributed to factors such as acidic pH, salt content, the presence of preservatives, emulsion characteristics, and nutrient deprivation. L. monocytogenes did not grow in six of the test products, but its population increased between 42 and 63 days in a butter-margarine blend stored at 10 degrees C and between 3 and 7 days when the blend was stored at 21 degrees C. On the basis of the experimental parameters examined in this study, traditional margarine and spreads not containing butter are not "potentially hazardous foods" in that they do not support the growth of Salmonella, E. coli O157:H7, or L. monocytogenes.     

Reduction of Escherichia coli O157:H7 and Salmonella enterica serovar Enteritidis in chicken manure by larvae of the black soldier fly

Green fluorescent protein-labeled Escherichia coli O157:H7 and Salmonella enterica serovar Enteritidis were inoculated at 10(7) CFU/g into cow, hog, or chicken manure. Ten- or 11-day-old soldier fly larvae (Hermetia illucens L.) (7 to 10 g) were added to the manure and held at 23, 27, or 32 degrees C for 3 to 6 days. Soldier fly larvae accelerated inactivation of E. coli O157:H7 in chicken manure but had no effect in cow manure and enhanced survival in hog manure. The initial pH values of the hog and chicken manure were 6.0 to 6.2 and 7.4 to 8.2, respectively, and it is surmised that these conditions affected the stability of the larval antimicrobial system. Reductions of E. coli O157:H7 populations in chicken manure by larvae were affected by storage temperature, with greater reductions in samples held for 3 days at 27 or 32 degrees C than at 23 degrees C. Pathogen inactivation in chicken manure by larvae was not affected by the indigenous microflora of chicken manure, because Salmonella Enteritidis populations in larvae-treated samples were approximately 2.5 log lower than control samples without larvae when either autoclaved or nonautoclaved chicken manure was used as the contaminated medium during 3 days of storage. Extending the storage time to 6 days, larvae again accelerated the reduction in Salmonella Enteritidis populations in chicken manure during the first 4 days of storage; however, larvae became contaminated with the pathogen. After 2 days of feeding on contaminated manure, Salmonella Enteritidis populations in larvae averaged 3.3 log CFU/g. Populations decreased to 1.9 log CFU/g after 6 days of exposure to contaminated chicken manure; however, the absence of feeding activity by the maggots in later stages of storage may be responsible for the continued presence of Salmonella Enteritidis in larvae. Transfer of contaminated larvae to fresh chicken manure restored feeding activity but led to cross-contamination of the fresh manure.     

Evaluation of a polymerase chain reaction-based system for detection of Salmonella enteritidis, Escherichia coli O157:H7, Listeria spp., and Listeria monocytogenes on fresh fruits and vegetables

A polymerase chain reaction (PCR)-based detection system, BAX, was evaluated for its sensitivity in detecting Salmonella Enteritidis, Escherichia coli O157:H7, Listeria sp., and Listeria monocytogenes on fresh produce. Fifteen different types of produce (alfalfa sprouts, green peppers, parsley, white cabbage, radishes, onions, carrots, mushrooms, leaf lettuce, tomatoes, strawberries, cantaloupe, mango, apples, and oranges) were inoculated, in separate studies, with Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes down to the predicted level of 1 CFU per 25-g sample. Detection by BAX was compared to recovery of the inoculated bacteria by culture methods according to the Food and Drug Administration's (FDA) Bacteriological Analytical Manual (BAM). BAX was essentially as sensitive as the culture-based method in detecting Salmonella Enteritidis and L. monocytogenes and more sensitive than the culture-based method for the detection of E. coli O157:H7 on green pepper, carrot, radish, and sprout samples. Detection of the pathogenic bacteria in samples spiked with a predicted number of less than 10 CFU was possible for most produce samples, but both methods failed to detect L. monocytogenes on carrot samples and one of two mushroom and onion samples spiked with less than 100 CFU. Both BAX and the culture method were also unable to consistently recover low numbers of E. coli O157:H7 from alfalfa sprouts. The PCR method allowed detection of Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes at least 2 days earlier than the conventional culture methods.     

Evaluation of universal preenrichment broth for growth of heat-injured pathogens.

Universal preenrichment broth (UPB) was developed to enable enrichment of injured foodborne pathogens of different genera simultaneously in lieu of having to undergo separate simultaneous enrichment cultures for subsequent detection or isolation of each pathogen. Enrichment conditions in UPB for growth of injured pathogens to populations that will enable pathogen detection by rapid immuno-based or polymerase chain reaction (PCR)-based assays have not been defined. Hence, studies were done to determine recovery and growth rates of heat-injured Escherichia coli O157:H7, Salmonella enterica ser. Typhimurium, Salmonella enterica ser. Enteritidis. and Listeria monocytogenes in UPB. Bacterial cells were heat injured in tryptic phosphate broth at 57.2 degrees C and inoculated at populations of ca. 0.17 to 63 injured cells per ml with raw ground beef, fresh chicken, lettuce, and environmental sponge samples. Enrichment cultures were sampled at 1, 2, 3, 4, 5, 6, and 24 h at 37 degrees C postinoculation, and pathogens were enumerated on appropriate selective media. Results revealed that recovery and growth of pathogens during the first 6 h of enrichment were not sufficient to ensure adequate numbers of bacteria (> 10(3) CFU/ ml) for detection by most immunoassays or PCR assays. Cells often required 3 to 4 h for recovery before growth was initiated. Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes cell populations in enrichment cultures with ground beef or lettuce at 6 h were 0.5 to 2.9 log10 CFU/ml. At 24 h of incubation, cell counts of enrichment samples for the three pathogens from all food and environmental sponge samples ranged from 4.0 to 8.3 log10 CFU/ml. Enrichment in UPB at 37 degrees C of foods or environmental sponge samples containing heat-injured cells of Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes reliably provides at 24 h of incubation-but not at 6 h-sufficient cell populations for detection by rapid immunoassay or PCR assay procedures that can detect at least 4 log10 CFU/ml. These results raise questions regarding the sensitivity of rapid detection methods that employ an abbreviated enrichment protocol of 6 h or less.     

Spread of bacterial pathogens during preparation of freshly squeezed orange juice

To study the potential of three bacterial pathogens to cross-contaminate orange juice during extraction, normal operation conditions during juice preparation at food service establishments were simulated. The spread of Salmonella enterica serovar Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes from inoculated oranges to work surfaces and to the final product was determined. The transference of these three bacterial pathogens to orange juice made from uninoculated oranges with the use of contaminated utensils was also studied. Fresh oranges were inoculated with a marker strain of rifampicin-resistant Salmonella Typhimurium, E. coli O157:H7, or L. monocytogenes. Final pathogen levels in juice were compared as a function of the use of electric or mechanical juice extractors to squeeze orange juice from inoculated oranges. Pathogen populations on different contact surfaces during orange juice extraction were determined on sulfite-phenol red-rifampicin plates for Salmonella Typhimurium and E. coli O157:H7 and on tryptic soy agar supplemented with 0.1 g of rifampicin per liter for L. monocytogenes. After inoculation, the average pathogen counts for the orange rind surface were 2.3 log10 CFU/cm2 for Salmonella Typhimurium, 3.6 log10 CFU/cm2 for E. coli O157:H7, and 4.4 log10 CFU/cm2 for L. monocytogenes. This contamination was spread over all utensils used in orange juice squeezing. Mean pathogen counts for the cutting board, the knife, and the extractor ranged from -0.3 to 2.1 log10 CFU/cm2, and the juice contained 1.0 log10 CFU of Salmonella Typhimurium per ml, 2.3 log10 CFU of E. coli O157:H7 per ml, and 2.7 log10 CFU of L. monocytogenes per ml. Contact with contaminated surfaces resulted in the presence of all pathogens in orange juice made from uninoculated oranges. These results give emphasis to the importance of fresh oranges as a source of pathogens in orange juice.     

Effects of recovery, plating, and inoculation methods on quantification of Escherichia coli O157:H7 and Listeria monocytogenes from strawberries

Effects of different recovery and inoculation methods on quantification of Escherichia coli O157:H7 and Listeria monocytogenes from strawberries were studied. Strawberries were spot or dip inoculated with 7 to 8 log CFU per strawberry of each pathogen, air dried for 2 h, and stored for 1, 3, and 7 days at 4 degrees C. The inoculated samples were stomached or washed with phosphate-buffered saline (PBS; pH 7.2) or with modified PBS (pH 8.4). Bacterial levels were determined using a direct selective plating, thin agar layer plating, or membrane-transferring plating (MTP) with tryptic soy agar and sorbital MacConkey agar (E. coli O157:H7) or modified Oxford agar (L. monocytogenes). Under most test conditions, washing with PBS followed by MTP had significantly higher (P < 0.05) recovery for both bacteria compared with other tested methods. Within a 7-day storage period for spot-inoculated strawberries, a stomaching step resulted in an injury of 0.9 to 1.4 log CFU for E. coli O157: H7 and 1.4 to 1.7 log CFU for L. monocytogenes. When a washing step was used instead, this resulted in an injury of only 0.2 to 0.6 log CFU for E. coli O157:H7 and 0.2 to 0.7 log CFU for L. monocytogenes. Both bacteria could survive on strawberry surfaces, but their recovered levels decreased with the increase of storage time at 4 degrees C for both spot and dip inoculation methods. Dip inoculation generally had a lower recovery than spot inoculation. An ideal protocol to recover and enumerate E. coli O157:H7 and L. monocytogenes from strawberries involved shaking and washing samples with 100 ml of PBS for 15 min at 22 degrees C coupled with a MTP enumeration method.     

Antimicrobial effect of herb extracts against Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium associated with beef

The effects of plant extracts against pathogenic bacteria in vitro are well known, yet few studies have addressed the effects of these compounds against pathogens associated with muscle foods. A series of experiments was conducted to determine the effectiveness of a commercially available, generally recognized as safe, herb extract dispersed in sodium citrate (Protecta One) or sodium chloride (Protecta Two) against Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes associated with beef. In the first experiment, E. coli O157:H7, Salmonella typhimurium, and L. monocytogenes inoculated onto beef and subjected to surface spray treatments with 2.5% solutions of Protecta One or Protecta Two were not affected by immediate application (day 0) of the herbal extracts. However, after 7 days of storage at 4 degrees C, E. coli O157:H7 was reduced by >1.3 log10 CFU/cm2 by Protecta Two; L. monocytogenes was reduced by 1.8 and 1.9 log10 CFU/cm2 by Protecta One and Protecta Two, respectively; Salmonella typhimurium was not reduced >0.3 log10 CFU/cm2 by either extract by day 7. In the second experiment, 2.5% Protecta Two (wt/vol or wt/wt) added to inoculated lean and adipose beef trim, processed, and packaged as ground beef chubs (80% lean, 20% adipose), did not reduce pathogen populations >0.5 log10 CFU/g up to 14 days at 4 degrees C. In the third experiment, surface spray treatments of beef with 2.5% lactic acid or 2.5% solutions of Protecta One or Protecta Two, vacuum packaged, and stored up to 35 days at 4 degrees C did reduce E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium slightly. These studies suggest that the use of herb extracts may afford some reductions of pathogens on beef surfaces; however, the antimicrobial activity may be diminished in ground beef by adipose components.     

Biodegradable polylactic acid polymer with nisin for use in antimicrobial food packaging

ABSTRACT:  Biodegradable polylactic acid (PLA) polymer was evaluated for its application as a material for antimicrobial food packaging. PLA films were incorporated with nisin to for control of foodborne pathogens. Antimicrobial activity of PLA/nisin films against Listeria monocytogenes , Escherichia coli O157:H7, and Salmonella Enteritidis were evaluated in culture media and liquid foods (orange juice and liquid egg white). Scanned electron micrograph and confocal laser microscopy revealed that nisin particles were evenly distributed in PLA polymer matrix on the surface and inside of the PLA/nisin films. PLA/nisin significantly inhibited growth of L . monocytogenes in culture medium and liquid egg white. The greatest inhibition occurred at 24 h when the cell counts of L. monocytogenes in the PLA/nisin samples were 4.5 log CFU/mL less than the controls. PLA/nisin reduced the cell population of E. coli O157:H7 in orange juice from 7.5 to 3.5 log at 72 h whereas the control remained at about 6 log CFU/mL. PLA/nisin treatment resulted in a 2 log reduction of S. Enteritidis in liquid egg white at 24 °C. After 21 d at 4 °C the S. Enteritidis population from PLA/nisin treated liquid egg white (3.5 log CFU/mL) was significantly less than the control (6.8 log CFU/mL). E. coli O157:H7 in orange juice was more sensitive to PLA/nisin treatments than in culture medium. The results of this research demonstrated the retention of nisin activity when incorporated into the PLA polymer and its antimicrobial effectiveness against foodborne pathogens. The combination of a biopolymer and natural bacteriocin has potential for use in antimicrobial food packaging.     

Death of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in shelf-stable, dairy-based, pourable salad dressings

The objectives of this study were to determine the death rates of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in three commercially manufactured full-fat ranch salad dressings, three reduced-fat ranch salad dressings, two full-fat blue cheese salad dressings, and two reduced-fat blue cheese salad dressings and to affirm the expectation that these dressings do not support the growth of these pathogens. The respective initial pH values of the four types of shelf-stable, dairy-based, pourable dressings were 2.87 to 3.72, 2.82 to 3.19, 3.08 to 3.87, and 2.83 to 3.49, respectively. Dressings were inoculated with low (2.4 to 2.5 log CFU/g) and high (5.3 to 5.9 log CFU/g) populations of separate five-strain mixtures of each pathogen and stored at 25 degrees C for up to 15 days. Regardless of the initial inoculum population, all test pathogens rapidly died in all salad dressings. Salmonella was undetectable by enrichment (<1 CFU/25-ml sample in three replicate trials) in all salad dressings within 1 day, and E. coli O157:H7 and L. monocytogenes were reduced to undetectable levels by enrichment between 1 and 8 days and 2 and 8 days, respectively. E. coli O157:H7 was not detected in 4 of the 10 salad dressings stored for 2 or more days and 9 of the 10 dressings stored for 6 or more days after inoculation. L. monocytogenes was detected in 9 of the 10 salad dressings stored for 3 days but in only one dressing, by enrichment, at 6 days, indicating that it had the highest tolerance among the three pathogens to the acidic environment imposed by the dressings. Overall, the type of dressing (i.e., ranch versus blue cheese) and level of fat in the dressings did not have a marked effect on the rate of inactivation of pathogens. Total counts and populations of lactic acid bacteria and yeasts and molds remained low or undetectable (<1.0 log CFU/ml) throughout the 15-day storage period. Based on these observations, shelf-stable, dairy-based, pourable ranch and blue cheese salad dressings manufactured by three companies and stored at 25 degrees C do not support the growth of Salmonella, E. coli O157:H7, and L. monocytogenes and should not be considered as potentially hazardous foods (time-temperature control for safety foods) as defined by the U.S. Food and Drug Administration Food Code.     

Inhibition of growth of pathogenic bacteria in raw milk by legume protein esters

Protein isolates from soybean and chickpea, as well as their methylated esters, were tested for their inhibitory action against the propagation of pathogenic bacteria in raw milk during its storage either at room temperature or under refrigeration. Raw milk was inoculated with a mixed culture of Listeria monocytogenes Scott A and Salmonella enterica serovar Enteritidis strain PT4 at ca. 2 log CFU ml?1. Aerobic plate count, coliform count, and presumptive E. coli in raw milk treated with esterified legume proteins were inhibited by 2 to 3 log relative to a control after 6 to 8 days of storage at 4°C. At room temperature, bacterial populations (aerobic plate count, coliform count, and presumptive E. coli) in raw milk treated with esterified legume proteins were inhibited by ca. 1.5 to 1.6 log relative to the control after 12 h. Supplementation of raw milk with esterified soybean protein could significantly inhibit the counts of the two inoculated pathogens (L. monocytogenes Scott A and Salmonella Enteritidis PT4), which were initially inoculated at ca. 2 log CFU ml?1, by ca. 2.4 log and 1.6 log CFU ml?1, respectively, on day 8 of storage under cold conditions. Corresponding reductions amounting to 2.7 and 1.8 log CFU ml?1 were observed after 12 h of storage at room temperature. Supplementation of raw milk with esterified soybean protein (0.5%) reduced the maximum level of titratable acidity to 0.21 and maintained the pH level at 6.4 after 8 days of storage under cold conditions as compared with 4 days for untreated raw milk. Similar results were observed when raw milk was stored at room temperature for 10 h.     

Inactivation of Salmonella on in-shell pecans during conditioning treatments preceding cracking and shelling

Studies were done to determine the effectiveness of conditioning treatments for killing Salmonella in and on immersion-inoculated and surface-inoculated in-shell pecans. Treatment of immersion-inoculated, dried, stored pecans in chlorinated water (400 μg/ml) reduced Salmonella by not more than 1.6 log CFU/g. Treatment of immersion-inoculated, dried, stored pecans in chlorinated water (200 μg/ml, 1 min) followed by soaking in water for 2 h at 21°C and treating for 10 min in water at 85 to 95°C reduced Salmonella by >5.12 log CFU/g; treatment of nuts containing a low population of Salmonella (<0.60 log CFU/g) for 15 min at 90°C failed to eliminate the pathogen. Reductions of ≥6.42 log CFU/g were achieved by treating surface-inoculated nuts in water at 90 or 95°C for 80 s; treatment of nuts containing 1.78 log CFU/g at 95°C for 10 min did not eliminate the pathogen. Salmonella on surface-inoculated in-shell pecans (kernel moisture, 4.75%; water activity, 0.62) that had been dried and stored at 4°C for 3 to 5 weeks was more resistant to conditioning treatments than was Salmonella on surface-inoculated pecans (kernel moisture, 5.60%; water activity, 0.73) that were not thoroughly dried. Conditioning treatments were less effective for killing Salmonella on immersion-inoculated pecans than on surface-inoculated pecans. Response of Salmonella to conditioning treatments varied, depending on the method of inoculation and whether nuts were dried and stored between the time of inoculation and treatment, which emphasizes the importance of following practices commonly used by commercial pecan shellers when validating the lethality of conditioning treatments.     

Efficacy of cetylpyridinium chloride in immersion treatment for reducing populations of pathogenic bacteria on fresh-cut vegetables.

The efficacy of cetylpyridinium chloride (CPC) immersion to reduce the numbers of three pathogenic bacteria (Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7) on three different fresh-cut vegetables (broccoli, cauliflower, and radishes) was studied. The fresh-cut vegetables were inoculated with one of the three pathogenic bacteria at a concentration of 10(5) CFU/ml for 1 h at room temperature and then treated with 0.1 or 0.5% CPC immersion for 1 min. Both Salmonella Typhimurium and E. coli O157:H7 plates were incubated from 48 to 72 h at 37 degrees C, and L. monocytogenes plates were incubated from 72 to 96 h before being counted. The results of three experiments showed that for the average of the three vegetables treated with 0.1 and 0.5% CPC, L. monocytogenes was reduced by 2.85 and 3.70 log CFU/g, Salmonella Typhimurium by 2.37 and 3.15 log CFU/g, and E. coli O157:H7 by 1.01 and 1.56 log CFU/g, respectively, in comparison with the vegetables treated with water only. The 0.5% CPC treatment was significantly different (P < 0.05) from the 0.1% CPC treatment on reduction of L. monocytogenes, Salmonella Typhimurium, and E. coli O157:H7. The CPC residual on the treated vegetables and their washing solutions were evaluated by using high-performance liquid chromatography.     

Survival of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella in juice concentrates

The survival of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella was studied in apple, orange, pineapple, and white grape juice concentrates and banana puree. Pouches of juice concentrate or puree were inoculated with pathogens at a level > or = 10(3) CFU/g and stored at -23 degrees C (-10 degrees F). Pathogen survival was monitored at 6 and 24 h, once a week for four consecutive weeks, and biweekly thereafter until 12 weeks. When pathogens were not detectable by direct plating, samples were enriched in universal preenrichment broth for 72 h and plated on selective media. Results showed that E. coli O157:H7, L. monocytogenes, and Salmonella were recoverable from all five concentrates through 12 weeks of storage at -23 degrees C.