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L R Beuchat

Publications and source records attributed to L R Beuchat.

At least 37 records · Page 2Linked to original sources

Factors influencing the detection and enumeration of Escherichia coli O157:H7 on alfalfa seeds.

Isolating Escherichia coli O157:H7 from batches of alfalfa seeds used to produce sprouts implicated in human illness has been difficult, perhaps due to nonhomogenous and very low-level contamination and inaccessibility of the pathogen entrapped in protected areas of the seed coat. We evaluated the effectiveness of various treatments in releasing E. coli O157:H7 from seeds. The influence of homogenization (blending or stomaching for 1 or 2 min), rinsing method (shaking for 5 min), soaking time (0. 1, 3, 6, or 15 h), soaking temperature (4 or 21 degrees C), and the addition of surfactants (0.1%, 0.5%, or 1.0% Tween 80 or Span 20) to rinse water was determined. Blending or stomaching for 1 or 2 min, and soaking for 1 h or longer at 4 or 21 degrees C, respectively, resulted in maximum release of E. coli O157:H7 from seeds. Soaking seeds at 37 degrees C for 15 h increased cell populations of E. coli O157:H7 by approximately 3.6 log10 CFU/g, likely due to bacterial growth. The maximum number of cells released from seeds by rinse water containing 1.0% Span 20 was at 21 degrees C, whereas at 37 degrees C, 0.1% or 0.5% Tween 80 was more effective. Detection of E. coli O157:H7 on seeds stored at 37 degrees C for up to 13 weeks and on sprouts derived from these seeds was compared. E. coli O157:H7 inoculated on seeds at 2.0 log10 CFU/g was detected after storage of seeds for up to 8 weeks at 37 degrees C and in sprouts produced from the seeds. The pathogen was not detected on seeds stored for 13 weeks at 37 degrees C and was not isolated from sprouts produced from these seeds. Identifying seed treatment methods that enhance removal of E. coli O157:H7 from alfalfa seeds can aid the isolation and enumeration of the pathogen on seeds. With a combination of optimal conditions for detecting the pathogen, i.e. soaking seeds for 1 h and pummeling seeds for 1 min, an enrichment step in modified tryptic soy broth (TSB), and the use of immunomagnetic beads for separation of E. coli O157:H7 cells, E. coli O157:H7 was detected in alfalfa seeds incubated at 37 degrees C for up to 8 weeks as effectively as in sprouts produced from the seeds.

Colony Count, Microbial↗

Performance of mycological media in enumerating desiccated food spoilage yeasts: an interlaboratory study.

Dichloran 18% glycerol agar (DG18) was originally formulated to enumerate nonfastidious xerophilic moulds in foods containing rapidly growing Eurotium species. Some laboratories are now using DG18 as a general purpose medium for enumerating yeasts and moulds, although its performance in recovering yeasts from dry foods has not been evaluated. An interlaboratory study compared DG18 with dichloran rose bengal chloramphenicol agar (DRBC), plate count agar supplemented with chloramphenicol (PCAC), tryptone glucose yeast extract chloramphenicol agar (TGYC), acidified potato dextrose agar (APDA), and orange serum agar (OSA) for their suitability to enumerate 14 species of lyophilized yeasts. The coefficient of variation for among-laboratories repeatability within yeast was 1.39% and reproducibility of counts among laboratories was 7.1%. The order of performance of media for recovering yeasts was TGYC > PCAC = OSA > APDA > DRBC > DG 18. A second study was done to determine the combined effects of storage time and temperature on viability of yeasts and suitability of media for recovery. Higher viability was retained at -18 degrees C than at 5 degrees C or 25 degrees C for up to 42 weeks, although the difference in mean counts of yeasts stored at -18 degrees C and 25 degrees C was only 0.78 log10 cfu/ml of rehydrated suspension. TGYC was equal to PCAC and superior to the other four media in recovering yeasts stored at -18 degrees C, 5 degrees C, or 25 degrees C for up to 42 weeks. Results from both the interlaboratory study and the storage study support the use of TGYC for enumerating desiccated yeasts. DG18 is not recommended as a general purpose medium for recovering yeasts from a desiccated condition.

Agar↗

Inhibitory activity of honey against foodborne pathogens as influenced by the presence of hydrogen peroxide and level of antioxidant power.

Antimicrobial activity of honey has been attributed to hydrogen peroxide, which is produced by naturally occurring glucose oxidase, and phenolic compounds, although lethality of and inhibition by these and other components against microorganisms vary greatly, depending on the floral source of nectar. This study was undertaken to compare honeys from six floral sources for their inhibitory activity against Escherichia coli O157:H7, Salmonella typhimurium, Shigella sonnei, Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus. A disc assay revealed that development of zones of inhibition of growth depends on the type and concentration of honey, as well as the test pathogen. Growth of B. cereus was least affected. The inhibition of growth of S. sonnei, L. monocytogenes, and S. aureus in 25% solutions of honeys was reduced by treating solutions with catalase, indicating that hydrogen peroxide contributes to antimicrobial activity. Darker colored honeys were generally more inhibitory than light colored honeys. Darker honeys also contained higher antioxidant power. Since antimicrobial activity of the darker colored test honeys was not eliminated by catalase treatment, non-peroxide components such as antioxidants may contribute to controlling the growth of some foodborne pathogens. The antibacterial properties of honeys containing hydrogen peroxide and characterized by a range of antioxidant power need to be validated using model food systems.

Anti-Bacterial Agents↗

Comparison of dichloran 18% glycerol (DG18) agar with general purpose mycological media for enumerating food spoilage yeasts.

Dichloran 18% glycerol (DG18) agar was originally developed to enumerate xerophilic foodborne moulds. However, some laboratories are using DG18 agar as a general medium to enumerate foodborne moulds and yeasts. A collaborative study, with the participation of seven laboratories, was undertaken to compare DG18 agar with dichloran rose bengal chloramphenicol (DRBC) agar, tryptone glucose yeast extract chloramphenicol (TGYC) agar, and plate count agar supplemented with chloramphenicol (PCAC) for enumerating 14 species of common food spoilage yeasts. Comparison of the mean values of populations of all yeasts recovered on each medium revealed no significant differences among DRBC agar, PCAC, and TGYC agar, while each of these media supported the development of significantly (P < or = 0.05) higher numbers of colonies than DG18 agar. However, differences were only 0.08 to 0.10 log10 cfu/ml, making the practical significance questionable. The overall coefficient of variation (CV) for within laboratory repeatability was 1.71%, while the CV for reproducibility of counts obtained among laboratories was 6.96%. Compared to DRBC agar, TGYC agar, and PCAC, yeast colonies were smaller on DG18 agar. Growth of Brettanomyces anomalus, Cryptococcus albidus, and Rhodotorula mucilaginosa was particularly retarded or inhibited on DG18 agar. Based on the performance of media in supporting colony development and ease of counting colonies, the use of DG18 agar as a general enumeration medium for foodborne yeasts cannot be recommended.

Agar↗

Effectiveness of immersion treatments with acids, trisodium phosphate, and herb decoctions in reducing populations of Yarrowia lipolytica and naturally occurring aerobic microorganisms on raw chicken.

Yarrowia lipolytica, one of the predominant yeasts in raw poultry, is believed to play a role in spoilage. This study was undertaken to investigate treatments to control the growth of Y. lipolytica on raw chicken stored at refrigeration temperature. Raw chicken wings inoculated with a mixture of five strains of Y. lipolytica isolated from raw poultry were dipped in solutions containing 2, 5, or 8% lactic acid, 2% lactic acid containing 0.2, 0.4, or 0.8% potassium sorbate or sodium benzoate, and 4, 8, or 12% trisodium phosphate solution. Populations of the yeast and total aerobic microorganisms were determined before and after treatment. Immersion of wings in 2% lactic acid (with or without 0.2% potassium sorbate or sodium benzoate) or 4% trisodium phosphate caused a significant (alpha = 0.05) reduction in numbers of Y. lipolytica and aerobic microorganisms. Treatment with 2% lactic acid containing 0.4 or 0.8% preservative did not result in additional significant reductions. Treatment of chicken wings with 2% lactic acid or 8% trisodium phosphate significantly reduced numbers of Y. lipolytica by 1.47 and 0.65 log10 cfu/g, respectively, and aerobic microorganisms by 2.60 and 1.21 log10 cfu/g, respectively, compared to controls. Growth of Y. lipolytica on wings stored at 5 degrees C for up to 9 days, however, was not affected by these treatments. Significant reductions in the population of Y. lipolytica occurred when the yeast was inoculated into 100% basil, marjoram, sage, and thyme decoctions, but not in 100% oregano or rosemary decoctions, held at 5 degrees C for 24 h. Treatment of chicken wings with 100% sage or thyme decoctions significantly reduced populations of Y. lipolytica but did not control its growth during storage at 5 degrees C for up to 9 days. The small, temporary decreases in numbers of Y. lipolytica and aerobic microorganisms resulting from immersion treatment of chicken wings with sage and thyme decoctions render these treatments of questionable value as preservation interventions.

Animals↗

Human pathogens associated with raw produce and unpasteurized juices, and difficulties in decontamination.

The ability of public health agencies to identify, through enhanced epidemiologic and surveillance techniques, raw fruits, vegetables, and unpasteurized juices as probable sources of infectious microorganisms, has undoubtedly resulted in increased numbers of documented outbreaks. Changes in agronomic, harvesting, distribution, processing, and consumption patterns and practices have also likely contributed to this increase. The risk of illness associated with raw produce and unpasteurized produce products can be reduced by controlling or preventing contamination, or by removing or killing pathogenic microorganisms by washing or treating them with sanitizers. However, the hydrophobic cutin, diverse surface morphologies, and abrasions in the epidermis of fruits and vegetables limit the efficacy of these treatments.

Food Handling↗

Survival of salmonellae on and in tomato plants from the time of inoculation at flowering and early stages of fruit development through fruit ripening.

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 for Salmonella. 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 which Salmonella may attach and remain viable during fruit development, thus serving as routes or reservoirs for contaminating ripened fruit.

Food Microbiology↗

Survival and heat resistance of Listeria monocytogenes after exposure to alkali and chlorine.

A strain of Listeria monocytogenes isolated from a drain in a food-processing plant was demonstrated, by determination of D values, to be more resistant to the lethal effect of heat at 56 or 59 degrees C following incubation for 45 min in tryptose phosphate broth (TPB) at pH 12.0 than to that of incubation for the same time in TPB at pH 7.3. Cells survived for at least 6 days when they were suspended in TPB at pHs 9.0, 10.0, and 11.0 and stored at 4 or 21 degrees C. Cells of L. monocytogenes incubated at 37 degrees C for 45 min and then stored for 48 or 144 h in TPB at pH 10.0 were more resistant to heat treatment at 56 degrees C than were cells stored in TPB at pH 7.3. The alkaline-stress response in L. monocytogenes may induce resistance to otherwise lethal thermal-processing conditions. Treatment of cells in 0.05 M potassium phosphate buffer (pH 7.00 +/- 0.05) containing 2.0 or 2.4 mg of free chlorine per liter reduced populations by as much as 1.3 log(10) CFU/ml, while treatment with 6.0 mg of free chlorine per liter reduced populations by as much as 4.02 log(10) CFU/ml. Remaining subpopulations of chlorine-treated cells exhibited some injury, and cells treated with chlorine for 10 min were more sensitive to heating at 56 degrees C than cells treated for 5 min. Contamination of foods by L. monocytogenes cells that have survived exposure to processing environments ineffectively cleaned or sanitized with alkaline detergents or disinfectants may have more severe implications than previously recognized. Alkaline-pH-induced cross-protection of L. monocytogenes against heat has the potential to enhance survival in minimally processed as well as in heat-and-serve foods and in foods on holding tables, in food service facilities, and in the home. Cells surviving exposure to chlorine, in contrast, are more sensitive to heat; thus, the effectiveness of thermal processing in achieving desired log(10)-unit reductions is not compromised in these cells.

Alkalies↗

Comparison of sample preparation methods for recovering Salmonella from raw fruits, vegetables, and herbs.

Methods for preparing raw fruits, vegetables, and herbs for enrichment or direct plating to determine the presence and populations of pathogenic bacteria vary greatly. A study was done to compare three sample processing methods (washing in 0.1% peptone, stomaching, and homogenizing) for their influence on recovery of Salmonella inoculated onto 26 types of raw produce. The mean numbers of Salmonella recovered from 10 fruits, 11 vegetables, and 5 herbs using all three processing methods were 7.17, 7.40, and 7.27 log10 CFU/sample, respectively. Considering all 26 types of produce and all processing methods, the number of Salmonella recovered ranged from 7.24 to 7.29 log10 CFU/sample, with no significant differences attributable to a particular sample processing method. Mean percent recoveries of Salmonella from washed, stomached, and homogenized produce were 39.4, 44.7, and 42.4%, respectively. Mean percent recoveries from fruits, vegetables, and herbs, regardless of sample preparation method, were 41.7, 50.1, and 25.9%, respectively. The number of Salmonella recovered from stomached and homogenized produce, but not washed produce, with pH < or = 4.53 was significantly less than the number recovered from produce with pH from 5.53 to 5.99, suggesting that the acidic environment in stomachates and homogenates was lethal to a portion of Salmonella. Reduced percent recoveries from herbs (pH 5.94 to 6.34) is attributed, in part, to antimicrobials released from plant cells during sample preparation. Overall, the type of processing method did not substantially affect the number of Salmonella recovered from the 26 types of raw produce representing a wide range of structural and morphological characteristics, composition, and pH. The influence of sample size, diluent composition, and processing time on efficiency of recovery of Salmonella and other pathogens needs to be evaluated before a method(s) for processing samples of raw produce can be recommended.

Colony Count, Microbial↗

Efficacy and reproducibility of a produce wash in killing Salmonella on the surface of tomatoes assessed with a proposed standard method for produce sanitizers.

The reproducibility of a method developed to evaluate point-of-use sanitizers for fresh produce was tested at three different laboratories. Mixtures of five Salmonella serotypes were inoculated on the surface of ripe tomatoes. After the inoculum was dry, tomatoes were placed inside a plastic bag and sprayed with sterile USP water, Dey and Engley (D/E) neutralizer broth, or a prototype Fit produce wash (PW), an alkaline solution comprised of generally recognized as safe ingredients (water, oleic acid, glycerol, ethanol, potassium hydroxide, sodium bicarbonate, citric acid, and distilled grapefruit oil), and rubbed for 30 s. The tomatoes were rinsed 10 s with 195 ml of D/E neutralizer broth (rinse solution), then combined with 20 ml of D/E neutralizer (residual wash solution) and rubbed by hand to remove residual Salmonella. Populations of Salmonella were determined for each tomato in the rinse solution and residual wash solution. Treatment with PW resulted in reductions in the number of Salmonella 2 to 4 logs greater than those achieved with the sterile water or D/E neutralizer broth controls. Consistent results were obtained across the three study sites, indicating reproducible results were obtained using the test method. The method used to determine the efficacy of killing or removing Salmonella from tomatoes in this study is suggested as a standard method for measuring the efficacy of sanitizers on tomatoes and other similar fruits and vegetables with rigid, smooth surfaces.

Colony Count, Microbial↗

Efficacy of chemical treatments in eliminating Salmonella and Escherichia coli O157:H7 on scarified and polished alfalfa seeds.

Alfalfa seeds are sometimes subjected to a scarification treatment to enhance water uptake, which results in more rapid and uniform germination during sprout production. It has been hypothesized that this mechanical abrasion treatment diminishes the efficacy of chemical treatments used to kill or remove pathogenic bacteria from seeds. A study was done to compare the effectiveness of chlorine (20,000 ppm), H2O, (8%), Ca(OH)2 (1%), Ca(OH)2 (1%) plus Tween 80 (1%), and Ca(OH)2 (1%) plus Span 20 (1%) treatments in killing Salmonella and Escherichia coli O157:H7 inoculated onto control, scarified, and polished alfalfa seeds obtained from two suppliers. The influence of the presence of organic material in the inoculum carrier on the efficacy of sanitizers was investigated. Overall, treatment with 1% Ca(OH)2 was the most effective in reducing populations of the pathogens. Reduction in populations of pathogens on seeds obtained from supplier I indicate that chemical treatments are less efficacious in eliminating the pathogens on scarified seeds compared to control seeds. However, the effectiveness of chemical treatment in removing Salmonella and E. coli O157:H7 from seeds obtained from supplier 2 was not markedly affected by scarification or polishing. The presence of organic material in the inoculum carrier did not have a marked influence on the efficacy of chemicals in reducing populations of test pathogens. Additional lots of control, scarified, and polished alfalfa seeds of additional varieties need to be tested before conclusions can be drawn concerning the impact of mechanical abrasion on the efficacy of chemical treatment in removing or killing Salmonella and E. coli O157:H7.

Calcium Hydroxide↗

Fate of salmonellae in calcium-supplemented orange juice at refrigeration temperature.

Recent outbreaks of salmonellosis associated with orange juice have raised interest concerning the survival and growth of Salmonella in juice supplemented with calcium. A study was done to determine the influence of various calcium supplements on the survival of salmonellae in orange juice held at 4 degrees C for up to 32 days. Isolates of Salmonellal Muenchen (inoculum 1), Salmonella isolates from humans and animals (inoculum 2), and Salmonella isolates from produce outbreaks (inoculum 3) were inoculated into pasteurized orange juices with pH values ranging from 3.96 to 4.19 and containing 350 mg of calcium per 240-ml serving (1.46 mg of calcium/ml). Populations of Salmonella declined rapidly in juice containing calcium lactate (CaL), with counts decreasing from 4.86 log10 CFU/ml to < 1 log10 CFU/ml within 16 days, regardless of the Salmonella serotypes present in inoculum. Counts decreased from 4.89 log10 CFU/ml to < 1 log10 CFU/ml in orange juice supplemented with CaL and tricalcium phosphate (TCP) within 30 days. These reductions were significantly (P < or = 0.05) higher than those of the control (no calcium added), in which Salmonella populations decreased 3.19 +/- 0.20 log10 CFU/ml over 32 days. Populations in orange juice containing TCP or calcium citrate (CC) declined 1.34 +/- 0.20 log10 CFU/ml and 1.96 +/- 0.20 log10 CFU/ml, respectively, over 32 days. These counts were significantly higher than respective control counts in juice stored for 32 days. Populations of Salmonella of inoculum 3 inoculated into juice containing calcium citrate malate (CCM) were significantly higher than in the control. Higher numbers of cells in inoculum 3 also survived compared to numbers of cells of inocula 1 or 2 in juice supplemented with CCM. This study reveals that the form of calcium used to supplement orange juice influences the ability of salmonellae to survive.

Beverages↗

Comparison of chlorine and a prototype produce wash product for effectiveness in killing Salmonella and Escherichia coli O157:H7 on alfalfa seeds.

Outbreaks of Salmonella and Escherichia coli O157:H7 infections associated with alfalfa and other seed sprouts have occurred with increased frequency in recent years. This study was undertaken to determine the efficacy of a liquid prototype produce wash product (Fit), compared with water and chlorinated water, in killing Salmonella and E. coli O157:H7 inoculated onto alfalfa seeds. We investigated the efficacy of treatments as influenced by seeds from two different lots obtained from two seeds suppliers and by two methods of inoculation. The efficacy of treatments was influenced by differences in seed lots and amount of organic material in the inoculum. Significant (alpha = 0.05) reductions in Salmonella populations on seeds treated with 20,000 ppm of chlorine or Fit for 30 min ranged from 2.3 to 2.5 log10 CFU/g and 1.7 to 2.3 log10 CFU/g, respectively. Reductions (alpha = 0.05) in E. coli O157:H7 ranged from 2.0 to 2.1 log10 CFU/g and 1.7 to more than 5.4 log10 CFU/g of seeds treated, respectively, with 20,000 ppm of chlorine or Fit. Compared with treatment with 200 ppm of chlorine, treatment with either 20,000 ppm of chlorine or Fit resulted in significantly higher reductions in populations of Salmonella and E. coli O157:H7. None of the treatments eliminated these pathogens as evidenced by their detection on enrichment of treated seeds. Considering the human health and environmental hazards associated with the use of 20,000 ppm of chlorine, Fit provides an effective alternative to chlorine as a treatment to significantly reduce bacterial pathogens that have been associated with alfalfa seeds.

Chlorine↗

Survival and growth of Escherichia coli O157:H7 inoculated onto cut lettuce before or after heating in chlorinated water, followed by storage at 5 or 15 degrees C.

This study determined the effects of mild heat and chlorine treatment followed by storage for up to 18 days at 5 degrees C or 7 days at 15 degrees C on the survival and growth of Escherichia coli O157:H7 inoculated onto fresh-cut iceberg lettuce. The efficacy of treatment with 20 ppm chlorine in killing the pathogen on lettuce at 50 degrees C was determined. Treatment of lettuce with 20 ppm chlorine at either 20 or 50 degrees C did not result in significantly greater reductions in populations of E. coli O157:H7 compared to respective treatments in water without chlorine. The pathogen steadily decreased in viability on treated lettuce throughout subsequent storage at 5 degrees C for 18 days. The population increased by 2.3 to 3.2 log10 CFU/g within 2 days, then continued to increase at a slower rate through 7 days of storage at 15 degrees C. At 4 and 7 days, significantly (alpha = 0.05) higher populations were reached on lettuce that had been treated at 50 degrees C, compared to respective samples that had been treated at 20 degrees C, regardless of the presence of 20 ppm chlorine in the treatment water. Treatment of lettuce with 20 ppm chlorine at 50 or 20 degrees C before or after inoculation with E. coli O157:H7 did not have a marked influence on behavior of the pathogen during subsequent storage at 5 or 15 degrees C.

Chlorine↗

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.

Colony Count, Microbial↗

Standardization of a method to determine the efficacy of sanitizers in inactivating human pathogenic microorganisms on raw fruits and vegetables.

The efficacy of sanitizers in killing human pathogenic microorganisms on a wide range of whole and fresh-cut fruits and vegetables has been studied extensively. Numerous challenge studies to determine the effects of storage conditions on survival and growth of pathogens on raw produce have also been reported. Results of these studies are often difficult to assess because of the lack of sufficient reporting of methods or, comparatively, because of variations in procedures for preparing and applying inocula to produce, conditions for treatment and storage, and procedures for enumerating pathogens. There is a need for a standard method to accurately determine the presence and populations of pathogenic microorganisms on produce. The adoption of standard, well-characterized reference strains would benefit a comparative assessment of a basic method among laboratories. A single protocol will not be suitable for all fruits and vegetables. Modifications of a basic method will be necessary to achieve maximum recovery of pathogens on various types of produce subjected to different sanitizer or storage treatments. This article discusses parameters that must be considered in the course of developing a basic standard method against which these modifications could be made.

Anti-Bacterial Agents↗

Sensitivity of acid-adapted and acid-shocked Shigella flexneri to reduced pH achieved with acetic, lactic, and propionic acids.

Survival and growth characteristics of unadapted, acid-adapted, and acid-shocked Shigella flexneri 2a cells in acidified (pH 3.5 to 5.5) tryptic soy broth with 0.25% glucose (TSB) and tryptic soy agar (TSA) were determined. S. flexneri was grown at 37 degrees C for 18 h in tryptic soy broth without glucose (TSBNG) (unadapted) and TSBNG supplemented with 1% glucose (TSBG) (acid-adapted). Cells grown in TSBNG were acid shocked by adjusting 16-h cultures to pH 5.05 +/- 0.05 with lactic acid. Cells were then inoculated into TSB acidified with acetic, lactic, or propionic acids to pH 5.5, 4.5, or 3.5 and incubated at 37 degrees C for 6 h. The order of lethality at a given pH was lactic acid < acetic acid < propionic acid. Significantly (P < or = 0.05) higher numbers of acid-adapted cells, compared to acid-shocked and unadapted cells, were recovered from TSB acidified (pH 3.5) with lactic or acetic acids. None of the cells survived a 30-min exposure in TSB acidified with propionic acid to pH 3.5. When the three cell types were plated on TSA acidified with lactic, acetic, or propionic acids at pH < or = 4.5, < or = 5.5, and < or = 5.5, respectively, visible colonies were not detected. Viable unadapted, acid-adapted, and acid-shocked cells were, however, recovered from TSA acidified with all three acids at pH > or = 4.5. Acid-adapted and, to a lesser extent, acid-shocked cells survived at lower pH than did unadapted cells, indicating that prior exposure to mild acidic environment results in increased acid resistance. Survival of S. flexneri at a given pH was influenced by the type of acidulant used, a response characteristic exhibited by other gram-negative enteric pathogens.

Acetic Acid↗

Development of a proposed standard method for assessing the efficacy of fresh produce sanitizers.

A series of studies was done for the purpose of developing a proposed standard method to evaluate point-of-use home sanitizers for fresh produce. Preliminary experiments were done to determine the survival of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes after inoculation onto the surface of ripe tomatoes and drying for up to 24 h at 22 +/- 2 degrees C. Within 2 h, the initial population (6.88 log10 CFU/tomato) of E. coli O157:H7 was reduced by approximately 3 log10, while reductions in similar initial populations of Salmonella and L. monocytogenes were approximately 1 and 0.6 log10 CFU/tomato, respectively, after 40 min and 3 h. A pilot study evaluated treatment with 200 ppm free chlorine and a prototype Fit produce wash (Fit) for their efficacy in killing a five-serotype mixture of Salmonella or L. monocytogenes spot inoculated on tomatoes using the proposed inoculation and recovery procedures. Inoculated tomatoes were sprayed with chlorinated water, Fit, or sterile distilled water (control) and hand rubbed for 30 s. Each tomato was then placed in a plastic bag and rinsed with 200 ml of sterile water by vigorously agitating for 30 s to simulate a procedure consumers might use for sanitizing and rinsing produce in a home setting. Each tomato was transferred to a second bag, and 20 ml of sterile 0.1% peptone was added; tomatoes were rubbed by hand for 40 s. Populations of Salmonella or L. monocytogenes in the rinse water and the 0.1% peptone wash solution were determined. Treatment with 200 ppm chlorine and Fit resulted in > or = 3.07 and > 6.83 log10 reductions, respectively, in Salmonella. Treatment with 200 ppm chlorine and Fit reduced the number of L. monocytogenes by > or = 3.33 and > or = 4.96 log10 CFU/tomato, respectively. The proposed standard method for testing the efficacy of point-of-use produce sanitizers needs to be evaluated for reproducibility of results through a larger scale series of experiments.

Chlorine↗