Synergistic effect of steam and lactic acid against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms on polyvinyl chloride and stainless steel

This study was designed to investigate the individual and combined effects of steam and lactic acid (LA) on the inactivation of biofilms formed by Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on polyvinyl chloride (PVC) and stainless steel. Six day old biofilms were developed on PVC and stainless steel coupons by using a mixture of three strains each of three foodborne pathogens at 25°C. After biofilm development, PVC and stainless steel coupons were treated with LA alone (immersed in 0.5% or 2% for 5s, 15s, and 30s), steam alone (on both sides for 5, 10, and 20s), and the combination of steam and LA. The numbers of biofilm cells of the three foodborne pathogens were significantly (p<0.05) reduced as the amount of LA and duration of steam exposure increased. There was a synergistic effect of steam and LA on the viability of biofilm cells of the three pathogens. For all biofilm cells of the three foodborne pathogens, reduction levels of individual treatments ranged from 0.11 to 2.12 log CFU/coupon. The combination treatment of steam and LA achieved an additional 0.2 to 2.11 log reduction compared to the sum of individual treatments. After a combined treatment of immersion in 2% LA for 15s or 30s followed by exposure to steam for 20s, biofilm cells of the three pathogens were reduced to below the detection limit (1.48 log). From the results of this study, bacterial populations of biofilms on PVC coupons did not receive the same thermal effect as on stainless steel coupons. Effectiveness of steam and LA may be attributed to the difference between Gram-negative and Gram-positive characteristics of the bacteria studied. The results of this study suggest that the combination of steam and LA has potential as a biofilm control intervention for food processing facilities.     

Inhibitory effects of UV treatment and a combination of UV and dry heat against pathogens on stainless steel and polypropylene surfaces

Abstract: Pathogens that contaminate the surfaces of food utensils may contribute to the occurrence of foodborne disease outbreaks. We investigated the efficacy of UV treatment combined with dry heat (50 °C) for inhibiting 5 foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Pseudomonas aeruginosa, Listeria monocytogenes, and Staphylococcus aureus) on stainless steel and polypropylene surfaces in this study. We inoculated substrates with each of the 5 foodborne pathogens cultured on agar surface and then UV treatment alone or a combination of both UV and dry heat (50 °C) was applied for 30 min, 1 h, 2 h, and 3 h. The initial populations of the 5 pathogens before treatment were 8.02 to 9.18 and 8.73 to 9.16 log₁₀ CFU/coupon on the surfaces of stainless steel and polypropylene coupons, respectively. UV treatments for 3 h significantly inhibited S. Typhimurium, L. monocytogenes, and S. aureus on the stainless steel by 3.06, 2.18, and 2.70 log₁₀ CFU/coupon, and S. aureus on the polypropylene by 3.11 log₁₀ CFU/coupon, respectively. The inhibitory effects of the combined UV and dry heat treatment (50 °C) increased as treatment time increased, yielding significant reductions in all samples treated for 3 h, with the exception of S. aureus on polypropylene. The reduction level of E. coli O157:H7 treated for 3 h on the surface of stainless steel and polypropylene treated was approximately 6.00 log₁₀ CFU/coupon. These results indicate that combined UV and dry heat (50 °C) treatments may be effective for controlling microbial contamination on utensils and cooking equipment surfaces as well as in other related environments.     

Inactivation of biofilm cells of foodborne pathogen by aerosolized sanitizers

The objective of this study was to determine the effect of aerosolized sanitizers on the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms. Biofilms were formed on a stainless steel and polyvinyl chloride (PVC) coupon by using a mixture of three strains each of three foodborne pathogens. Six day old biofilms on stainless steel and PVC coupons were treated with aerosolized sodium hypochlorite (SHC; 100 ppm) and peracetic acid (100, 200, and 400 ppm) in a model cabinet for 5, 10, 30, and 50 min. Treatment with 100 ppm PAA was more effective than the same concentration of SHC with increasing treatment time. Exposure to 100 ppm SHC and PAA for 50 min significantly (p<0.05) reduced biofilm cells of three foodborne pathogens (0.50 to 3.63 log CFU/coupon and 2.83 to more than 5.78 log CFU/coupon, respectively) compared to the control treatment. Exposure to 200 and 400 ppm PAA was more effective in reducing biofilm cells. Biofilm cells were reduced to below the detection limit (1.48 log CFU/coupon) between 10 and 30 min of exposure. The results of this study suggest that aerosolized sanitizers have a potential as a biofilm control method in the food industry.     

Inactivation of Escherichia coli O157:H7 on stainless steel upon exposure to Paenibacillus polymyxa biofilms

We investigated the potential use of biofilm formed by a competitive-exclusion (CE) microorganism to inactivate Escherichia coli O157:H7 on a stainless steel surface. Five microorganisms showing inhibitory activities against E. coli O157:H7 were isolated from vegetable seeds and sprouts. The microorganism with the greatest antimicrobial activity was identified as Paenibacillus polymyxa (strain T5). In tryptic soy broth (TSB), strain T5 reached a higher population at 25 °C than at 12 or 37 °C without losing inhibitory activity against E. coli O157:H7. When P. polymyxa (6 log CFU/mL) was co-cultured with E. coli O157:H7 (2, 3, 4, or 5 log CFU/mL) in TSB at 25 °C, the number of E. coli O157:H7 decreased significantly within 24 h. P. polymyxa formed a biofilm on stainless steel coupons (SSCs) in TSB at 25 °C within 24 h, and cells in biofilms, compared to attached cells without biofilm formation, showed significantly increased resistance to a dry environment (43% relative humidity [RH]). With the exception of an inoculum of 4 log CFU/coupon at 100% RH, upon exposure to biofilm formed by P. polymyxa on SSCs, populations of E. coli O157:H7 (2, 4, or 6 log CFU/coupon) were significantly reduced within 48 h. Most notably, when E. coli O157:H7 at 2 log CFU/coupon was applied to SSCs on which P. polymyxa biofilm had formed, it was inactivated within 1 h, regardless of RH. These results will be useful when developing strategies using biofilms produced by competitive exclusion microorganisms to inactivate foodborne pathogens in food processing environments.     

Resistance of pathogenic bacteria on the surface of stainless steel depending on attachment form and efficacy of chemical sanitizers

Various bacteria including food spoilage bacteria and pathogens can form biofilms on different food processing surfaces, leading to potential food contamination or spoilage. Therefore, the survival of foodborne pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, Cronobacter sakazakii) in different forms (adhered cells, biofilm producing in TSB, biofilm producing at RH 100%) on the surface of stainless steel and stored at various relative humidities (RH 23%, 43%, 68%, 85%, and 100%) at room temperature for 5 days was investigated in this study. Additionally, the efficacy of chemical sanitizers (chlorine-based and alcohol-based commercial sanitizers) on inhibiting various types of biofilms of E. coli O157:H7 and S. aureus on the surface of stainless steel was investigated. The number of pathogens on the surface of stainless steel in TSB stored at 25 °C for 7 days or RH 100% at 25 °C for 7 days was significantly increased and resulted in the increase of 3 log₁₀ CFU/coupon after 1 day, and these levels were maintained for 7 days. When stainless steel coupons were stored at 25 °C for 5 days, the number of pathogens on the surface of stainless steel was significantly reduced after storage at RH 23%, 43%, 68%, and 85%, but not at 100%. When the bacteria formed biofilms on the surface of stainless steel in TSB after 6 days, the results were similar to those of the attached form. However, levels of S. aureus and C. sakazakii biofilms were more slowly reduced after storage at RH 23%, 43%, 68%, and 85% for 5 days than were those of the other pathogens. Formation of biofilms stored at RH 100% for 5 days displayed the highest levels of resistance to inactivation. Treatment with the alcohol sanitizer was very effective at inactivating attached pathogens or biofilms on the surface of stainless steel. Reduction levels of alcohol sanitizer treatment ranged from 1.91 to 4.77 log and from 4.35 to 5.35 log CFU/coupon in E. coli O157:H7 and S. aureus, respectively. From these results, the survival of pathogens contaminating the surfaces of food processing substrates such as stainless steel varied depending on RH and attachment form. Also, alcohol-based sanitizers can be used as a potential method to remove microbial contamination on the surfaces of utensils, cooking equipment, and other related substrates regardless of the microbial attached form.     

Behavior of Escherichia coli O157:H7 and Listeria monocytogenes in Tryptic soy broth subjected to various low temperature treatments

The inactivation and injury of Escherichia coli O157:H7 and Listeria monocytogenes in Tryptic soy broth stored at −5, −18 and −28°C were studied. Regardless of storage temperature, viable populations of E. coli O157:H7 and L. monocytogenes determined with TSA (uninjured and injured cells) or TSAB (uninjured cells), decreased as the storage time increased. However, the least surviving population of both test organisms was noted when stored at −18°C followed by those stored at −28 and −5°C. The viable populations of E. coli O157:H7 determined either with TSA or TSAB, was reduced most drastically during the first day of storage then decreased slowly thereafter. Viable populations of L. monocytogenes declined slightly and gradually during the entire storage period. Furthermore, E. coli O157:H7 was found more susceptible to the freezing storage than L. monocytogenes. After 21-day storage at −18°C, population reduction of E. coli O157:H7 determined with TSA was ca 1.72 log CFU/ml. On the other hand, a population reduction of only 0.64 log CFU/ml was noted with L. monocytogenes. Besides, the surviving population of E. coli O157:H7 contained a larger proportion of injured cells than L. monocytogenes.     

Fate of biofilm cells of Cronobacter sakazakii under modified atmosphere conditions

Survival of biofilm cells of Cronobacter sakazakii formed on stainless steel and polyvinyl chloride (PVC) on exposure to different atmosphere conditions was studied. Biofilms were formed on stainless steel and PVC coupons by using three strains of C. sakazakii. Six day old biofilms on stainless steel and PVC coupons were stored under N₂ gas, CO₂ gas, and air for up to 20 days. N₂ and CO₂ gases resulted in significant (p < 0.05) further reductions of 1.79 and 2.47 log CFU/cm² after 20 days of storage, respectively, compared to air storage. N₂ and CO₂ gases led to less reduction of biofilm cells on PVC compared to those on stainless steel. N₂ and CO₂ gases resulted in significant (p < 0.05) further reductions of 0.98 and 1.20 log CFU/cm² after 20 days of storage, respectively, compared to air storage.     

Magnetic nano-beads based separation combined with propidium monoazide treatment and multiplex PCR assay for simultaneous detection of viable Salmonella Typhimurium, Escherichia coli O157:H7 and Listeria monocytogenes in food products

We developed a rapid and reliable technique for simultaneous detection of Salmonella Typhimurium, Escherichia coli O157:H7 and Listeria monocytogenes that can be used in food products. Magnetic nano-beads (MNBs) based immunomagnetic separation (IMS) was used to separate the target bacterial cells while multiplex PCR (mPCR) was used to amplify the target genes. To detect only the viable bacteria, propidium monoazide (PMA) was applied to selectively suppress the DNA detection from dead cells. The results showed the detection limit of IMS-PMA-mPCR assay was about 10² CFU/ml (1.2 × 10² CFU/ml for S. Typhimurium, 4.0 × 10² CFU/ml for E. coli O157:H7 and 5.4 × 10² CFU/ml for L. monocytogenes) in pure culture and 10³ CFU/g (5.1 × 10³ CFU/g for S. Typhimurium, 7.5 × 10³ CFU/g for E. coli O157:H7 and 8.4 × 10³ CFU/g for L. monocytogenes) in spiking food products samples (lettuce, tomato and ground beef). This report has demonstrated for the first time, the effective use of rapid and reliable IMS combined with PMA treatment and mPCR assay for simultaneous detection of viable S. Typhimurium, E. coli O157:H7 and L. monocytogenes in spiked food samples. It is anticipated that the present approach will be applicable to simultaneous detection of the three target microorganisms for practical use.     

Assessment of bacterial biofilm on stainless steel by hyperspectral fluorescence imaging

Hyperspectral fluorescence imaging techniques were investigated for detection of two genera of microbial biofilms on stainless steel material which is commonly used to manufacture food processing equipment. Stainless steel coupons were deposited in nonpathogenic E. coli O157:H7 and Salmonella cultures, prepared using M9 minimal medium with casamino acids (M9C), for 6 days at 37 °C. Hyperspectral fluorescence emission images of the biofilm formations on the stainless coupons were acquired from 416 to 700 nm with the use of ultraviolet-A (320–400 nm) excitation. In general, emission peaks for both bacteria were observed in the blue region at approximately 480 nm and thus provided the highest contrast between the biofilms and background stainless steel coupons. A simple thresholding of the 480 nm image showed significantly larger biofilm regions for E. coli O157:H7 than for Salmonella. Viable cell counts suggested that Salmonella formed significantly higher density biofilm regions than E. coli O157:H7 in M9C medium. On the basis of principal component analysis (PCA) of the hyperspectral fluorescence images, the second principal component image exhibited the most distinguishable morphological differences for the concentrated biofilm formations between E. coli and Salmonella. E. coli formed granular aggregates of biofilms above the medium on stainless steel while Salmonella formed dense biofilm in the medium-air interface region (pellicle). This investigation demonstrated the feasibility of implementing fluorescence imaging techniques to rapidly screen large surface areas of food processing equipment for bacterial contamination. Company and product names are used for clarity and do not imply any endorsement by USDA to the exclusion of other comparable products.     

Antibacterial activities of a cinnamon essential oil with cetylpyridinium chloride emulsion against <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 and <Emphasis Type="Italic">Salmonella</Emphasis> Typhimurium in basil leaves

This study examined the antibacterial activities of two different cinnamon essential oil emulsions against Escherichia coli O157:H7 and Salmonella Typhimurium on basil leaves. Cinnamon oil (0.25%) treatments containing CPC (0.05%) exhibited greater effects on the pathogenic bacteria than cinnamon oil treatment without this emulsifier (p < 0.05). Treatment with cinnamon bark and leaf oil emulsions (CBE and CLE, respectively) reduced the populations of E. coli O157:H7 by 4.10 and 5.10 log CFU/g, and S. Typhimurium by 2.71 and 2.82 log CFU/g, respectively. Scanning electron micrographs showed morphological changes in the two pathogenic bacteria following emulsion treatment. In addition, there was no difference in the color or ascorbic acid content of the basil leaves by the emulsion treatment. These results suggest that CBE or CLE treatment can be an effective way to ensure the microbial safety of minimally processed vegetables and a good alternative to chlorination treatment in the fresh produce industry.     

The Effect of pH on Inactivation of Pathogenic Bacteria on Fresh-cut Lettuce by Dipping Treatment with Electrolyzed Water

Fresh‐cut lettuce samples inoculated with S. Typhimurium, E. coli O157:H7 or L. monocytogenes were dipped into 300 ppm electrolyzed water (EW) at pH 4 to 9 and 30 °C for 5 min. The effects of treatment pH on bacterial reduction and visual quality of the lettuce were determined. The treatments at pH 4 and 8 resulted in the most effective inactivation of E. coli O157:H7, but the effect of pH was not significant (P > 0.05) for S. Typhimurium and L. monocytogenes. The treatment at pH 7 retained the best visual quality of lettuce, and achieved a reduction of approximately 2 log CFU/g for above 3 bacteria.     

Attachment and biofilm formation by Escherichia coli O157:H7 at different temperatures, on various food-contact surfaces encountered in beef processing

Escherichia coli O157:H7 attached to beef-contact surfaces found in beef fabrication facilities may serve as a source of cross-contamination. This study evaluated E. coli O157:H7 attachment, survival and growth on food-contact surfaces under simulated beef processing conditions. Stainless steel and high-density polyethylene surfaces (2 × 5 cm) were individually suspended into each of three substrates inoculated (6 log CFU/ml or g) with E. coli O157:H7 (rifampicin-resistant, six-strain composite) and then incubated (168 h) statically at 4 or 15 °C. The three tested soiling substrates included sterile tryptic soy broth (TSB), unsterilized beef fat-lean tissue (1:1 [wt/wt]) homogenate (10% [wt/wt] with sterile distilled water) and unsterilized ground beef. Initial adherence/attachment of E. coli O157:H7 (0.9 to 2.9 log CFU/cm²) on stainless steel and high-density polyethylene was not affected by the type of food-contact surface but was greater (p < 0.05) through ground beef. Adherent and suspended E. coli O157:H7 counts increased during storage at 15 °C (168 h) by 2.2 to 5.4 log CFU/cm² and 1.0 to 2.8 log CFU/ml or g, respectively. At 4 °C (168 h), although pathogen levels decreased slightly in the substrates, numbers of adherent cells remained constant on coupons in ground beef (2.4 to 2.5 log CFU/cm²) and increased on coupons in TSB and fat-lean tissue homogenate by 0.9 to 1.0 and 1.7 to 2.0 log CFU/cm², respectively, suggesting further cell attachment. The results of this study indicate that E. coli O157:H7 attachment to beef-contact surfaces was influenced by the type of soiling substrate and temperature. Notably, attachment occurred not only at a temperature representative of beef fabrication areas during non-production hours (15 °C), but also during cold storage (4 °C) temperatures, thus, rendering the design of more effective sanitation programs necessary.     

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes in milk by caprylic acid and monocaprylin

Listeria monocytogenes and Escherichia coli O157:H7 are major foodborne pathogens implicated in various outbreaks involving pasteurized or unpasteurized milk, and various dairy products. The objective of this study was to determine the antibacterial effect of caprylic acid (CA, C_8:0) and its monoglyceride, monocaprylin (MC) on L. monocytogenes and E. coli O157:H7 in whole milk. A five-strain mixture of E. coli O157:H7 or L. monocytogenes was inoculated in autoclaved milk (10⁶ CFU/ml) containing 0, 25, or 50 mM of CA or MC. At 37°C, all the treatments, excepting 25 mm CA, reduced the population of both pathogens by approximately 5.0 log CFU/ml in 6 h. At 24 h of storage at 8°C, MC at both levels and CA at 50 mM decreased L. monocytogenes and E. coli O157:H7, respectively by >5.0 log CFU/ml. At 48 h of 4°C storage, populations of L. monocytogenes and E. coli O157:H7 were decreased to below detection level (enrichment negative) by 50 mm of MC and CA, respectively. Results indicate that MC could potentially be used to inhibit L. monocytogenes and E. coli O157:H7 in milk and dairy products, but sensory studies need to be conducted before recommending their use.     

Behavior of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium in teewurst, a raw spreadable sausage

The fate of Listeria monocytogenes, Salmonella Typhimurium, or Escherichia coli O157:H7 were separately monitored both in and on teewurst, a traditional raw and spreadable sausage of Germanic origin. Multi-strain cocktails of each pathogen (ca. 5.0 log CFU/g) were used to separately inoculate teewurst that was subsequently stored at 1.5, 4, 10, and 21 °C. When inoculated into commercially-prepared batter just prior to stuffing, in general, the higher the storage temperature, the greater the lethality. Depending on the storage temperature, pathogen levels in the batter decreased by 2.3 to 3.4, ca. 3.8, and 2.2 to 3.6 log CFU/g for E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, during storage for 30 days. When inoculated onto both the top and bottom faces of sliced commercially-prepared finished product, the results for all four temperatures showed a decrease of 0.9 to 1.4, 1.4 to 1.8, and 2.2 to 3.0 log CFU/g for E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, over the course of 21 days. With the possible exceptions for salt and carbohydrate levels, chemical analyses of teewurst purchased from five commercial manufacturers revealed only subtle differences in proximate composition for this product type. Our data establish that teewurst does not provide a favourable environment for the survival of E. coli O157:H7, S. Typhimurium, or L. monocytogenes inoculated either into or onto the product.     

Reduction of Escherichia coli O157:H7 in Biofilms Using Bacteriophage BPECO 19

Biofilm formation is a growing concern in the food industry. Escherichia coli O157:H7 is one of the most important foodborne pathogens that can persists in food and food‐related environments and subsequently produce biofilms. The efficacy of bacteriophage BPECO 19 was evaluated against three E. coli O157:H7 strains in biofilms. Biofilms of the three E. coli O157:H7 strains were grown on abiotic (stainless steel, rubber, and minimum biofilm eradication concentration [MBEC^TM] device) and biotic (lettuce) surfaces at different temperatures. The effectiveness of bacteriophage BPECO 19 in reducing preformed biofilms on these surfaces was further evaluated by treating the surfaces with a phage suspension (10⁸ PFU/mL) for 2 h. The results indicated that the phage treatment significantly reduced (P  < 0.05) the number of adhered cells in all the surfaces. Following phage treatment, the viability of adhered cells was reduced by ≥3 log CFU/cm², 2.4 log CFU/cm², and 3.1 log CFU/peg in biofilms grown on stainless steel, rubber, and the MBEC^TM device, respectively. Likewise, the phage treatment reduced cell viability by ≥2 log CFU/cm² in biofilms grown on lettuce. Overall, these results suggested that bacteriophages such as BPECO 19 could be effective in reducing the viability of biofilm‐adhered cells.     

Rhizosphere effect on survival of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in manure-amended soil during cabbage (Brassica oleracea) cultivation under tropical field conditions in Sub-Saharan Africa

The effect of cabbage (Brassica oleracea) rhizosphere on survival of Escherichia coli O157:H7 and Salmonella Typhimurium in manure-amended soils under tropical field conditions was investigated in the Central Agro-Ecological Zone of Uganda. Three-week old cabbage seedlings were transplanted and cultivated for 120 days on manure-amended soil inoculated with 4 or 7 log CFU/g non-virulent E. coli O157:H7 and S. Typhimurium. Cabbage rhizosphere did not affect survival of the 4 log CFU/g inocula in manure-amended soil and the two enteric bacteria were not detected on/in cabbage leaves at harvest. The 7 log CFU/g E. coli O157:H7 and S. Typhimurium survived in bulk soil for a maximum of 80 and 96 days, respectively, but the organisms remained culturable in cabbage rhizosphere up to the time of harvest. At 7 log CFU/g inoculum, E. coli O157:H7 and S. Typhimurium contamination on cabbage leaves occurred throughout the cultivation period. Leaf surface sterilisation with 1% AgNO₃ indicated that the organisms were present superficially and in protected locations on the leaves. These results demonstrate that under tropical field conditions, cabbage rhizosphere enhances the persistence of E. coli O157:H7 and S. Typhimurium in manure-amended soil at high inoculum density and is associated with long-term contamination of the leaves.     

Antimicrobial Activities of Phenolic Extracts Derived from Seed Coats of Selected Soybean Varieties

Soybean hulls or seed coats consist of complex carbohydrates, proteins, lipids, and polyphenols such as anthocyanidins, proanthocyanidins, and isoflavones. The polyphenolics in the seed coats give them various colors such as black, brown, green, yellow, or even a mottled appearance. In this study, the antimicrobial effects of phenolic extracts from the seed coats of different colored soybeans (yellow, dark brown, brown, and black) were evaluated against foodborne pathogens such as Salmonella Typhimurium, Escherichia coli O157:H7, and Campylobacter jejuni in broth‐cultures as well as on chicken skin. The highest total phenolic content was observed for the phenolic extract from soybean variety (R07‐1927) with black colored seed coat (74.1 ± 2.1 mg chlorogenic acid equivalent [CAE]/g extract) and was significantly different (P <0.0001) from the extract of the conventional soybean variety (R08‐4004) with yellow colored seed coat (7.4 ± 1.2 mg CAE/g extract). The extract from black colored soybean produced reductions of 2.10 ± 0.08 to 2.20 ± 0.08‐log CFU/mL for both E. coli O157:H7 and C. jejuni after 3 d when incubated in broth‐culture having 4‐log CFU/mL of bacteria, whereas a 6 d incubation was found to reduce S. Typhimurium and E. coli O157:H7 at 2.03 ± 0.05 and 3.3 ± 0.08‐log CFU/mL, respectively. The extract also reduced S. Typhimurium and E. coli O157:H7 attached to chicken skin by 1.39 ± 0.03 and 1.24 ± 0.06‐log CFU/g, respectively, upon incubation for 6 d. Soybean seed coat extracts may have a potency as antimicrobial agents to reduce foodborne bacteria contaminating poultry products.     

Biofilm formation and sporulation by Bacillus cereus on a stainless steel surface and subsequent resistance of vegetative cells and spores to chlorine, chlorine dioxide, and a peroxyacetic acid-based sanitizer

Biofilm formation by Bacillus cereus 038-2 on stainless steel coupons, sporulation in the biofilm as affected by nutrient availability, temperature, and relative humidity, and the resistance of vegetative cells and spores in biofilm to sanitizers were investigated. Total counts in biofilm formed on coupons immersed in tryptic soy broth (TSB) at 12 and 22 degrees C consisted of 99.94% of vegetative cells and 0.06% of spores. Coupons on which biofilm had formed were immersed in TSB or exposed to air with 100, 97, 93, or 85% relative humidity. Biofilm on coupons immersed in TSB at 12 degrees C for an additional 6 days or 22 degrees C for an additional 4 days contained 0.30 and 0.02% of spores, respectively, whereas biofilm exposed to air with 100 or 97% relative humidity at 22 degrees C for 4 days contained 10 and 2.5% of spores, respectively. Sporulation did not occur in biofilm exposed to 93 or 85% relative humidity at 22 degrees C. Treatment of biofilm on coupons that had been immersed in TSB at 22 degrees C with chlorine (50 microg/ml), chlorine dioxide (50 microg/ml), and a peroxyacetic acid-based sanitizer (Tsunami 200, 40 microg/ml) for 5 min reduced total cell counts (vegetative cells plus spores) by 4.7, 3.0, and 3.8 log CFU per coupon, respectively; total cell counts in biofilm exposed to air with 100% relative humidity were reduced by 1.5, 2.4, and 1.1 log CFU per coupon, respectively, reflecting the presence of lower numbers of vegetative cells. Spores that survived treatment with chlorine dioxide had reduced resistance to heat. It is concluded that exposure of biofilm formed by B. cereus exposed to air at high relative humidity (> or =97%) promotes the production of spores. Spores and, to a lesser extent, vegetative cells embedded in biofilm are protected against inactivation by sanitizers. Results provide new insights to developing strategies to achieve more effective sanitation programs to minimize risks associated with B. cereus in biofilm formed on food contact surfaces and on foods.     

Production of biofilm and quorum sensing by Escherichia coli O157:H7 and its transfer from contact surfaces to meat,poultry, ready-to-eat deli,and produce products

Multistate outbreaks of Escherichia coli O157:H7 infections through consumption of contaminated foods including produce products have brought a great safety concern. The objectives of this study were to determine the effect of biofilm and quorum sensing production on the attachment of E. coli O157:H7 on food contact surfaces and to evaluate the transfer of the pathogen from the food contact to various food products. E. coli O157:H7 produced maximum levels of AI-2 signals in 12 h of incubation in tested meat, poultry, and produce broths and subsequently formed strong biofilm in 24 h of incubation. In general, E. coli O157:H7 formed stronger biofilm on stainless steel than glass. Furthermore, E. coli O157:H7 that had attached on the surface of stainless steel was able to transfer to meat, poultry, ready-to-eat deli, and produce products. Strong attachment of the transferred pathogen on produce products (cantaloupe, lettuce, carrot, and spinach) was detected (>10³ CFU/cm²) even after washing these products with water. Our findings suggest that biofilm formation by E. coli O157:H7 on food contact surfaces can be a concern for efficient control of the pathogen particularly in produce products that require no heating or cooking prior to consumption.     

Predicting process lethality of Escherichia coli O157:H7, Salmonella,and Listeria monocytogenes in ground,formulated, and formed beef/turkey links cooked in an air impingement oven

The pathogen thermal lethality in ground and formulated beef/turkey was evaluated for a cocktail of E. coli O157:H7, Salmonella, and Listeria monocytogenes, respectively. At a temperature range of 55–70°C, the heat resistance of L. monocytogenes was not significantly (at α=0.05) different from those of Salmonella. The heat resistance of L. monocytogenes at 55–70°C was 45–81% higher than that of E. coli O157:H7. In this study, a practical approach was developed to predict log₁₀(CFU/g) reduction of E. coli O157:H7, Salmonella, or L. monocytogenes in ground, formulated, and formed beef/turkey links that were cooked in an air impingement oven. The predictions of pathogen thermal kills in the links were verified via the inoculation studies for at least a 7 log₁₀(CFU/g) reduction of E. coli O157:H7, Salmonella, and L. monocytogenes.