Identification of the genetic elements involved in biofilm formation by Salmonella enterica serovar Tennessee using mini-Tn10 mutagenesis and DNA sequencing.

Biofilms are known to contribute to bacterial tolerance to desiccation and survival in low moisture foods. However, the molecular mechanisms underlying biofilm formation have not been fully elucidated. This study identified some of the genes that are implicated in biofilm formation by Salmonella enterica serovar Tennessee, the "peanut butter outbreak" strain. Mini-Tn10 mutagenesis was used in the study to generate random transposon insertion libraries. The ability of selected mutants in forming biofilms was compared with their wildtype parent using the crystal violet binding assay. Mutants forming significantly less (P ≤ 0.05) biofilm compared to their wildtype parent were selected for whole-genome sequencing. Mini-Tn10 insertion sites on mutant genomes were identified by comparing the acquired sequencing data with those in the Genbank using the BLAST search. In total, 56 mutants were obtained, and five were selected for further analysis according to the result of the biofilm assay. Sequencing analysis revealed that the mini-Tn10 interrupted the S. enterica genes that encode bacterial cell membrane lipoprotein, DNA topoisomerase III, attachment and invasion locus protein, bacteriocin immunity protein, and cell division protein. The information generated from the research should be useful in the control fo S. enterica in low-moisture foods and their production environments.

Efficacy of commercial overhead washing and waxing systems on the microbiological quality of fresh peaches.

Overhead spray washing and waxing systems (WWS) are used commercially to reduce the risk of microbial contamination and improve the quality of fresh produce during packing. This study evaluated the microbiological quality of overhead spray water and spent peach wash water, as well as fresh peaches before and after they pass the WWS. Pre- and post-washed/waxed peach samples (n = 192) and overhead spray water and spent peach wash water samples (n = 54) were collected several times over the course of a processing day in three packing facilities located in the state of Georgia. Populations of total aerobes (TA), yeasts and molds (YM), and coliforms (TC) and the presence of thermotolerant coliforms (TTC) and enterococci (EC) were measured in collected samples. The average TA and TC counts and the incidences of TTC and EC were significantly higher (P < 0.05) on peach samples collected after the WWS compared to those collected before the WWS. Counts and incidences of TA, YM, and TC in spent peach wash water were significantly higher than in the overhead spray water where neither TTC nor EC was detected. Results suggest that the commercial washing and waxing systems had little effect in improving the microbiological quality of fresh peaches.

Interactions between fish isolates Pseudomonas fluorescens and Staphylococcus aureus in dual-species biofilms and sensitivity to carvacrol.

Biofilm formation is a frequent source of contamination of food products, which results in significant economic losses through microbial spoilage and poses serious health concerns. Little is known about the fate of Staphylococcus aureus in the dual-species biofilms with Pseudomonas fluorescens an important spoiler commonly found in aquatic products. This study evaluates the interactions between mono- or dual-species biofilms formed by P. fluorescens and S. aureus, as well as the sensitivity of the two tested strains to carvacrol. The biofilm cell population, expolysaccharide production, biofilm structures of P. fluorescens as mono- and dual-species with S. aureus at ratios of 1:1 and 1:0.01 were investigated with different concentrations of carvacrol (0, 0.4, 0.8 and 1.6 mM) in fish juice at 30 °C. The results show that the biofilm cell population of S. aureus in the dual-species was significantly lower (p < 0.05) than that in the mono-species, compared to no difference for P. fluorescens. In the co-culture the dominance of P. fluorescens inhibited the growing population of S. aureus in both planktonic and biofilm cells, however, two strains were stimulated to produce the large expolysaccharides and coaggregation, forming the complex spatial multibiofilm structures. The large increase in the dual-species biofilms was positively correlated with high quorum sensing autoinductor-2 (AI-2), and exogenous 4,5-dihydroxy-2,3-pentanedione (the AI-2 precursor, DPD), rather than C4-HSL, greatly stimulated the dual-species biofilm formation. In addition, carvacrol significantly reduced the tested biofilms and expolysaccharide secretion without affecting cell viability in a concentration-dependent manner, especially for S. aureus. Furthermore, the two strains as the dual-species biofilms exhibited lower sensitivity to carvacrol than the mono-culture, regardless of the level of inoculum of S. aureus, which was consistent with the decrease of AI-2 activity. The present study highlights that the interactions between P. fluorescens and S. aureus in dual-species biofilms promoted the large production of expolysaccharides and complex biofilm structures modulated by AI-2 signal, which results in the community-level resistance to carvacrol.

Microencapsulation of Lactobacillus plantarum NRRL B-1927 with Skim Milk Processed via Ultra-High-Pressure Homogenization.

Several health benefits are associated with the consumption of probiotic foods. Lyophilized probiotic cultures are commonly used to manufacture probiotic-containing products. Spray drying (SDR) is a cost-effective process to microencapsulate probiotics. However, the high temperatures of the drying air in SDR can inactivate significant numbers of probiotic cells. Ultra-high-pressure homogenization (UHPH) processing can modify the configuration of proteins found in skim milk which may increase its protective properties as microencapsulating agent towards probiotic cells during SDR. The aim of this study was to evaluate the effect of microencapsulating probiotic Lactobacillus plantarum NRRL B-1927 (LP) with UHPH-treated skim milk after SDR or freeze drying (FD). Dispersions containing LP were made with either UHPH-treated (at 150 MPa or 300 MPa) or untreated skim milk and dried via concurrent SDR (CCSD), mixed-flow SDR (MXSD) or FD. Higher cell survival (%) of LP was found in powders microencapsulated with 150 MPa-treated skim milk than in those microencapsulated with non-UHPH-treated and 300 MPa-treated skim milk via FD followed by MXSD and CCSD, respectively. Increasing UHPH pressures increased the particle size of powders produced via CCSD; and reduced particle agglomeration of powders produced via MXSD and FD. This study demonstrated that UHPH processes improves the effectiveness of skim milk as a microencapsulating agent for LP, creating powders that could be used in probiotic foods.

Fate of Salmonella enterica and Enterohemorrhagic Escherichia coli Cells Artificially Internalized into Vegetable Seeds during Germination.

Vegetable seeds contaminated with bacterial pathogens have been linked to fresh-produce-associated outbreaks of gastrointestinal infections. This study was undertaken to observe the physiological behavior of Salmonella enterica and enterohemorrhagic Escherichia coli (EHEC) cells artificially internalized into vegetable seeds during the germination process. Surface-decontaminated seeds of alfalfa, fenugreek, lettuce, and tomato were vacuum-infiltrated with four individual strains of Salmonella or EHEC. Contaminated seeds were germinated at 25°C for 9 days, and different sprout/seedling tissues were microbiologically analyzed every other day. The internalization of Salmonella and EHEC cells into vegetable seeds was confirmed by the absence of pathogens in seed-rinsing water and the presence of pathogens in seed homogenates after postinternalization seed surface decontamination. Results show that 317 (62%) and 343 (67%) of the 512 collected sprout/seedling tissue samples were positive for Salmonella and EHEC, respectively. The average Salmonella populations were significantly larger (P < 0.05) than the EHEC populations. Significantly larger Salmonella populations were recovered from the cotyledon and seed coat tissues, followed by the root tissues, but the mean EHEC populations from all sampled tissue sections were statistically similar, except in pregerminated seeds. Three Salmonella and two EHEC strains had significantly larger cell populations on sprout/seedling tissues than other strains used in the study. Salmonella and EHEC populations from fenugreek and alfalfa tissues were significantly larger than those from tomato and lettuce tissues. The study showed the fate of internalized human pathogens on germinating vegetable seeds and sprout/seedling tissues and emphasized the importance of using pathogen-free seeds for sprout production.IMPORTANCE The internalization of microorganisms into vegetable seeds could occur naturally and represents a possible pathway of vegetable seed contamination by human pathogens. The present study investigated the ability of two important bacterial pathogens, Salmonella and enterohemorrhagic Escherichia coli (EHEC), when artificially internalized into vegetable seeds, to grow and disseminate along vegetable sprouts/seedlings during germination. The data from the study revealed that the pathogen cells artificially internalized into vegetable seeds caused the contamination of different tissues of sprouts/seedlings and that pathogen growth on germinating seeds is bacterial species and vegetable seed-type dependent. These results further stress the necessity of using pathogen-free vegetable seeds for edible sprout production.

Differential Attachment of Salmonella enterica and Enterohemorrhagic Escherichia coli to Alfalfa, Fenugreek, Lettuce, and Tomato Seeds.

Vegetable seeds have the potential to disseminate and transmit foodborne bacterial pathogens. This study was undertaken to assess the abilities of selected Salmonella and enterohemorrhagic Escherichia coli (EHEC) strains to attach to fungicide-treated versus untreated, and intact versus mechanically damaged, seeds of alfalfa, fenugreek, lettuce, and tomato. Surface-sanitized seeds (2 g) were exposed to four individual strains of Salmonella or EHEC at 20°C for 5 h. Contaminated seeds were rinsed twice, each with 10 ml of sterilized water, before being soaked overnight in 5 ml of phosphate-buffered saline at 4°C. The seeds were then vortexed vigorously for 1 min, and pathogen populations in seed rinse water and soaking buffer were determined using a standard plate count assay. In general, the Salmonella cells had higher attachment ratios than the EHEC cells. Lettuce seeds by unit weight had the highest numbers of attached Salmonella or EHEC cells, followed by tomato, alfalfa, and fenugreek seeds. In contrast, individual fenugreek seeds had more attached pathogen cells, followed by lettuce, alfalfa, and tomato seeds. Significantly more Salmonella and EHEC cells attached to mechanically damaged seeds than to intact seeds (P < 0.05). Although, on average, significantly more Salmonella and EHEC cells were recovered from untreated than fungicide-treated seeds (P < 0.05), fungicide treatment did not significantly affect the attachment of individual bacterial strains to vegetable seeds (P > 0.05), with a few exceptions. This study fills gaps in the current body of literature and helps explain bacterial interactions with vegetable seeds with differing surface characteristics.IMPORTANCE Vegetable seeds, specifically sprout seeds, have the potential to disseminate and transmit foodborne bacterial pathogens. This study investigated the interaction between two important bacterial pathogens, i.e., Salmonella and EHEC, and vegetable seeds with differing surface characteristics. This research helps understand whether seed surface structure, integrity, and fungicide treatment affect the interaction between bacterial cells and vegetable seeds.

Isolation and characterisation of Bacillus spp. antagonistic to Vibrio parahaemolyticus for use as probiotics in aquaculture.

Acute gastroenteritis caused by pathogenic Vibrio parahaemolyticus is one of the major factors affecting the development of aquaculture and the safety of seafood. Using the antagonism of probiotics against pathogens is an alternative strategy to antibiotics and a common trend to control food-borne pathogenic bacteria. In this study, a total of 249 isolates were isolated from four types of seafood (Litopenaeus vannamei, Oratosquilla oratoria, Mactra veneriformis and Portunus trituberculatus) and coastal sediment from Liaodong Bay in the Bohai Sea, China with five different separation agars. The most isolates came from the sample of coastal sediment and on agar of 2216E, which accounted for 36.14 and 54.62 % respectively. Twenty-four among 249 isolates displayed direct antimicrobial activity to V. parahaemolyticus with spot inoculation. Sixteen active isolates were selected for extracellular antimicrobial activity using the Oxford cup method. Only strains of B16 and J7 showed extracellular antimicrobial activity and were identified as Bacillus pumilus and Bacillus mojavensis respectively based on the physiological identification and 16S rRNA sequence analysis. Both of the strains B16 and J7 exhibited extracellular hydrolytic enzyme activity and antagonism against more than one indicator bacteria in vitro, which indicates that the two strains have broad potential application as suitable probiotic candidates in aquaculture while B. mojavensis was first reported to inhibit pathogenic Vibrio spp. in vitro. There is no particular trait as to antagonism of B. pumilus B16 or B. mojavensis J7 to Gram-positive or Gram-negative indicator bacteria.

Survival of four commercial probiotic mixtures in full fat and reduced fat peanut butter.

A well-documented health benefit of probiotics is their ability to reduce the incidence of diarrhea in young, malnourished children in the developing countries. This study was undertaken to determine whether peanut butter, a nutritious, low-moisture food could be a carrier for probiotics by observing the survivability of selected probiotic mixtures in peanut butter under different storage conditions. Commercial probiotic mixtures (B, U, N and S) comprising of multiple strains of Lactobacillus, Bifidobacterium, Streptococcus and Lactococcus were inoculated into full fat or reduced fat peanut butter at 10(7) CFU/g. Resulting products were stored at 4, 25 or 37 °C for 12 months. Populations of Lactobacillus, Bifidobacterium and Streptococcus/Lactococcus were determined periodically. The average viable cell counts of N and S were significantly lower than those of B and U (p < 0.05). In all probiotic products stored at different temperatures, Bifidobacterium had the greatest survivability, followed by Lactobacillus and Streptococcus/Lactococcus. The probiotics used in the study had different surviving patterns, and their survival was influenced by storage conditions. Fat content of peanut butter had no significant impacts on probiotic viability. Results suggest that peanut butter can be a vehicle to deliver probiotics for preventing diarrhea among malnourished children.

[Quantitative classification-based occupational health management for electroplating enterprises in Baoan District of Shenzhen, China].

OBJECTIVE: To improve the occupational health management levels in electroplating enterprises with quantitative classification measures and to provide a scientific basis for the prevention and control of occupational hazards in electroplating enterprises and the protection of workers' health. METHODS: A quantitative classification table was created for the occupational health management in electroplating enterprises. The evaluation indicators included 6 items and 27 sub-items, with a total score of 100 points. Forty electroplating enterprises were selected and scored according to the quantitative classification table. These electroplating enterprises were classified into grades A, B, and C based on the scores. RESULTS: Among 40 electroplating enterprises, 11 (27.5%) had scores of >85 points (grade A), 23 (57.5%) had scores of 60∼85 points (grade B), and 6 (15.0%) had scores of <60 points (grade C). CONCLUSION: Quantitative classification management for electroplating enterprises is a valuable attempt, which is helpful for the supervision and management by the health department and provides an effective method for the self-management of enterprises.

The fate of enterohemorrhagic Escherichia coli on alfalfa and fenugreek seeds and sprouts as affected by ascaroside #18 treatments.

Alfalfa and fenugreek sprouts are healthy foods, but they are occasionally contaminated with bacterial pathogens and serve as vehicles for transmitting foodborne illnesses. This study examined the efficacy of ascaroside (ascr)#18 treatment for the control of enterohemorrhagic E. coli (EHEC) growth on sprouts. Commercial alfalfa and fenugreek seeds were decontaminated with 20,000 ppm of NaClO, and residual chlorine was neutralized with Dey-Engley broth. Decontaminated seeds were treated with 1 mM or 1 µM ascr#18, a plant immunity modulator, before being dried and mixed with sandy soil inoculated with E. coli F4546 or BAA-2326 at 104-105 CFU/g. The inoculated seeds were sprouted on 1% water agar at 25ºC for 7 days in the dark. Seed or sprout samples were collected on days 0, 1, 3, 5, and 7 for enumeration of bacterial populations. Data was fit into the general linear model and analyzed using Fisher's least significant different test of the statistical analysis software. Treatment with ascr#18 significantly (P ≤ 0.05) reduced the cell population of EHEC on sprouts. The mean EHEC populations in the 1 mM or 1 µM treatment groups were 3.31 or 1.56 log CFU/g lower compared to the control groups. Besides treatment, sprout seed type and sprouting time were also significant independent variables influencing the growth of EHEC, according to the results of type III error analysis. However, EHEC strain type was not a significant independent variable. The study suggests that ascr#18 could be potentially used to control EHEC contamination and improve the microbial safety of sprouts.

Pesticides residues in leafy green vegetables and irrigation waters in Accra, Ghana.

Pesticides are used in vegetable farming to control pests and diseases, reduce crop losses and improve yield. The study examined pesticide residues in irrigation waters and leafy green vegetables grown in some farming areas in Accra, Ghana. Three types of irrigation water sources (n = 23) and two exotic and four indigenous Ghanaian leafy vegetables (n = 34) from 10 farming areas in Accra, Ghana were collected and examined for 15 organochlorines, 13 organophosphorus and 9 synthetic pyrethroids pesticide residues using the modified QuEChERS procedure. Pesticide residues were detected on 50% (17/34) of the leafy vegetable and 52% (12/23) of the irrigation water samples analysed. Chlorpyrifos and deltamethrin were the most detected pesticide residues in the vegetables and irrigation water. About 26.5% of the vegetables contained pesticide residues exceeding the EU maximum residue limits, so vegetable farmers should be encouraged to comply with appropriate measures on pesticide use to enhance food safety.

Microencapsulated polyphenol extracts from Georgia-grown pomegranate peels delay lipid oxidation in salad dressing during accelerated and ambient storage conditions.

Lipid oxidation is a major cause of quality deterioration in salad dressings. This study evaluated the effect of incorporating microencapsulated polyphenol extracts via spray drying from pomegranate peels (MPP) to delay lipid oxidation in Italian-style salad dressings (ISD) during accelerated (55°C) and ambient (25°C) storage conditions. ISDs, prepared at high (5000 rpm) and low (250 rpm) shear rates conditions, were formulated with unencapsulated polyphenol extracts from pomegranate peels (PPP), MPP, and/or grape seed extract (GSE). Lipid oxidation in ISDs was evaluated by measuring peroxide value (PV), iodine value (IV), and TBARS, stored in accelerated and ambient conditions for 21 days and 8 weeks, respectively. Tannis in extracts were measured via HPLC-DAD and the total hydrolyzable tannin content of PPP and MPP was 283.09 and 427.74 (mg/g extract), respectively. Condensed tannins were not detected in PPP and MPP but were found in GSE (348.53 mg/g extract). Salad dressings prepared at high shear rates had significantly (p < .05) higher emulsion stability than those homogenized at low shear rates. Mixing conditions did not affect the lipid oxidative stability of IDSs. Salad dressing stored under accelerated storage had higher lipid oxidation (higher PV, lower IV, and higher TBARS) after 21 days than IDSs stored under ambient conditions for 8 weeks. ISDs prepared with MPPP showed significantly (p < .05) lower lipid oxidation than the other ISDs at the end of the shelf life studies. Results from the accelerated storage suggested that incorporating MPP could have extended the shelf life of IDSs by 20% compared to using unencapsulated polyphenol extracts. The study demonstrated that MPP delays lipid oxidation in ISDs during storage more effectively than unencapsulated extracts. MPP may serve as a natural and effective functional food ingredient for controlling lipid oxidation in high-lipid and acidified foods.

Ellagitannin content and anti-enterohemorrhagic Escherichia coli activity of aqueous extracts derived from commercial pomegranate products.

This study compared the efficacy of aqueous extracts of commercially available pomegranate peel products and a juice powder in inhibiting the growth of two enterohemorrhagic Escherichia coli strains. Cell suspension of each E. coli strain (5 Log CFU/ml) was added into tryptic soy broth amended with 9 or 23% of each extract prepared with two different methods. After treatment for 5, 10, and 24 h at 25 °C, surviving E. coli cells were enumerated on tryptic soy agar to determine cell population reduction compared to the controls. The concentrations of six different ellagitannins and titratable activity in each treatment system were determined and correlated to E. coli cell population reduction. The extracts from three powdered pomegranate peels caused a significantly greater (p ≤ 0.05) reduction in E. coli population than the extract from the whole peel and juice powder. The higher dose of extracts resulted in a greater cell population reduction than the lower dose. The level of E. coli population reduction correlated positively with the total ellagitannins content (R2 0.67-0.98) and the titratable acidity (R2 0.69-0.98) in the treatment systems. The study suggests that pomegranate peels are promising natural additives or preservatives to control pathogens like EHEC.

Formulation of a drinkable peanut-based therapeutic food for malnourished children using plant sources.

High ingredient costs continue to hamper local production of therapeutic foods (TFs). Development of formulations without milk, the most expensive ingredient, is one way of reducing cost. This study formulated a ready-to-drink peanut-based TF that matched the nutrient composition of F100 using plant sources. Three least cost formulations namely, A, B and C were designed using computer formulation software with peanuts, beans, sesame, cowpeas and grain amaranth as ingredients. A 100 g portion of the TF provided 101-111 kcal, 5 g protein and 5.3-6.5 g fat. Consumer acceptability hedonic tests showed that the products were liked (extremely and moderately) by 62-65% of mothers. These results suggest that nutrient dense TFs formulated from only plant sources have the potential to be used in the rehabilitation phase of the management of malnourished children after clinical testing.

Efficacy of Ascaroside #18 Treatments in Control of Salmonella enterica on Alfalfa and Fenugreek Seeds and Sprouts.

A novel, natural, and effective antimicrobial intervention is in demand for improving the microbial safety of vegetable seeds/sprouts. This study assessed the efficacy of ascaroside treatment in the control of Salmonella enterica on alfalfa and fenugreek sprouts. Sanitized commercial seeds were treated with 1 mM or 1 µM ascaroside (ascr)#18, a plant immunity modulator (PIM) and dried for an hour before being inoculated with lyophilized S. Cubana or S. Stanley cells in sandy soil (104 CFU/g). Treated and untreated seeds were spouted on 1% water agar at 25°C in the dark. Seed or sprout samples were collected on days 0, 1, 3, 5, and 7, and the population of Salmonella was determined. Data were fit into the general linear arrangement, and means were separated using Fisher's least significant difference test. Seed type, strain type, treatment type, and sprouting time were significant factors (P ≤ 0.05) influencing Salmonella growth on sprouts. The populations of Salmonella were significantly higher on fenugreek than on alfalfa sprouts. S. Stanley had a significantly higher population than S. Cubana. The population of Salmonella increased from day 0 to day 3 and reached the peak population on Day 5. Treatments with both concentrations of ascaroside significantly decreased the populations of Salmonella compared to the controls. The mean Salmonella population reduction was ca. 4 or 1 log CFU/g by treatment with 1 mM and 1 µM of the PIM, respectively. Treatment with the PIM could be potentially used to improve the microbial safety of vegetable seeds and sprouts.

Microbial Load of Fresh Blueberries Harvested by Different Methods.

Currently, more and more growers are transitioning to the use of over-the-row machine harvesters for harvesting fresh market blueberries. This study assessed the microbial load of fresh blueberries harvested by different methods. Samples (n = 336) of 'Draper' and 'Liberty' northern highbush blueberries, which were harvested using a conventional over-the-row machine harvester, a modified machine harvester prototype, ungloved but sanitized hands, and hands wearing sterile gloves were collected from a blueberry farm near Lynden, WA, in the Pacific Northwest at 9 am, 12 noon, and 3 pm on four different harvest days during the 2019 harvest season. Eight replicates of each sample were collected at each sampling point and evaluated for the populations of total aerobes (TA), total yeasts and molds (YM), and total coliforms (TC), as well as for the incidence of fecal coliforms and enterococci. The harvest method was a significant factor (p < 0.05) influencing the TA and TC counts, the harvest time was a significant factor influencing the YM counts, while the blueberry cultivar was an insignificant (p > 0.05) factor for all three indicator microorganisms. These results suggest that effective harvester cleaning methods should be developed to prevent fresh blueberry contamination by microorganisms. This research will likely benefit blueberry and other fresh fruit producers.

Application of multiple occupational health risk assessment models in occupation health risk prediction of trichloroethylene in the electroplating and electronics industries.

Objectives. This study assessed the occupational health risks of work group exposure to trichloroethylene (TCE) in the electroplating and electronics industries in China. Methods. The UK Control of Substances Hazardous to Health (COSHH) Essential, the US Environmental Protection Agency (EPA) and the Singapore and the Chinese semiquantitative risk assessment models were used to assess the risks of TCE. Twenty degreasing groups and 14 cleaning groups were recruited in the companies selected. Results. The concentrations of TCE in 66.7% of the cleaning groups and 35.0% of the degreasing groups exceeded the permissible concentration time-weighted average (PC-TWA) in China, and the concentrations of TCE in 100.0% of the cleaning groups and 70.0% of the degreasing groups exceeded the permissible concentration short-term exposure limit (PC-STEL) in China. Over 60.0% of the work groups were evaluated at high risk and over half of the work groups were evaluated at high cancer risk by the risk assessment models. Conclusion. Most work groups exposed to TCE in the electroplating and electronics industries in China are at high risk. The cleaning groups may have a higher risk for TCE exposure. The Chinese exposure index method and the synthesis index method are more practical than the other methods.

Validating the Efficacy of Sanitation Methods Commonly Used by Ghanaian Households in Inactivating Artificially Inoculated Salmonella enterica on Leafy Green Vegetables.

ABSTRACT: Our previous survey revealed the poor microbial quality of leafy green vegetables and the presence of Salmonella on these vegetables grown and sold in Accra, Ghana. This study validated the efficacy of some cleaning and sanitation methods (tap water, salt solution, lemon juice, and vinegar) commonly used by Ghanaian households, by comparing them with the performance of several sanitation approaches used by the U.S. fresh produce industry (chlorine, peracetic acid, and ozonated water) in reducing Salmonella populations on leafy green vegetables. Cabbage and lettuce leaves inoculated with each of three Salmonella cocktails were treated with sterile water and the previously mentioned six sanitizers. The efficacies of the treatments were evaluated by using the standard plate count assay. The effect of the treatments on the sensory quality of treated vegetables was evaluated by a 97-member consumer panel in the United States. Treatments with citric and acetic acid were as effective as chlorine and peracetic acid in reducing Salmonella counts on vegetable leaves. Ozonated water was less effective than the two organic acids but equally as effective as treatment with the salt solution. Rinsing vegetables with water did not significantly reduce Salmonella counts (P > 0.05). Cabbage leaves treated with citric acid, vinegar, and water were more preferred (P ≤ 0.05) by the consumer panel, while chlorine-treated cabbage leaves were the least preferred. Lettuce samples treated with citric acid and water were more preferred, and salt-treated samples were least preferred. Among the vegetable sanitation methods used by Ghanaian households, treatments with citric and acetic acid are effective in inactivating Salmonella without adversely affecting the sensory quality of treated vegetables.

Effect of Pre-Exposure to Chlorine Dioxide on the Susceptibility of Fecal Coliforms to Antibiotics.

Adaptive exposure to sub-lethal concentrations of sanitizers was previously reported to offer cross-protection to bacteria against antibiotics. This study was undertaken to determine whether the pre-exposure of fecal coliforms to suboptimal concentrations of a chemical sanitizer, chlorine dioxide (ClO2), alters their susceptibility to certain antibiotics. Fecal coliforms isolated from fresh fruit packing facilities (n = 12) were adapted in ½ or » of the manufacturer-recommended concentration of ClO2. The susceptibility of the adapted and non-adapted cells to 13 different antibiotics was determined by observing the changes in their minimal inhibitory concentrations (MICs). The results showed that preadaptation to the suboptimal concentrations of ClO2, in general, either decreased or did not change the MICs of the antibiotics against selected fecal coliform isolates, with only two exceptions; preadaptation increased the MICs of kanamycin against two of the fecal coliform isolates, and of nalidixic acid against one of the fecal coliform isolates. The results suggest that the use of ClO2 has a relatively low risk of inducing the resistance of fecal coliforms to antibiotics.

Inactivation of shiga toxin-producing Escherichia coli (STEC) and degradation and removal of cellulose from STEC surfaces by using selected enzymatic and chemical treatments.

Some Shiga toxin-producing Escherichia coli (STEC) strains produce extracellular cellulose, a long polymer of glucose with ß-1-4 glycosidic bonds. This study evaluated the efficacies of selected enzymatic and chemical treatments in inactivating STEC and degrading/removing the cellulose on STEC surfaces. Six cellulose-producing STEC strains were treated with cellulase (0.51 to 3.83 U/15 ml), acetic and lactic acids (2 and 4%), as well as an acidic and alkaline sanitizer (manufacturers' recommended concentrations) under appropriate conditions. Following each treatment, residual amounts of cellulose and surviving populations of STEC were determined. Treatments with acetic and lactic acids significantly (P < 0.05) reduced the populations of STEC, and those with lactic acid also significantly decreased the amounts of cellulose on STEC. The residual amounts of cellulose on STEC positively correlated to the surviving populations of STEC after the treatments with the organic acids (r = 0.64 to 0.94), and the significance of the correlations ranged from 83 to 99%. Treatments with cellulase and the sanitizers both degraded cellulose. However, treatments with cellulase had no influence on the fate of STEC, and those with the sanitizers reduced STEC cell populations to undetectable levels. Thus, the correlations between the residual amounts of cellulose and the surviving populations of STEC caused by these two treatments were not observed. The results suggest that the selected enzymatic and chemical agents degraded and removed the cellulose on STEC surfaces, and the treatments with organic acids and sanitizers also inactivated STEC cells. The amounts of cellulose produced by STEC strains appear to affect their susceptibilities to certain sanitizing treatments.