scholarly journals Biofilm Formation by Shiga Toxin-Producing Escherichia coli on Stainless Steel Coupons as Affected by Temperature and Incubation Time

2019 ◽  
Vol 7 (4) ◽  
pp. 95 ◽  
Author(s):  
Zhi Ma ◽  
Emmanuel W. Bumunang ◽  
Kim Stanford ◽  
Xiaomei Bie ◽  
Yan D. Niu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Shiga Toxin ◽  
Incubation Time ◽  
Food Processing ◽  
Biofilm Formation ◽  
Crystal Violet Staining ◽  
Plate Counts ◽  
Air Liquid Interface ◽  
Over Time

Forming biofilm is a strategy utilized by Shiga toxin-producing Escherichia coli (STEC) to survive and persist in food processing environments. We investigated the biofilm-forming potential of STEC strains from 10 clinically important serogroups on stainless steel at 22 °C or 13 °C after 24, 48, and 72 h of incubation. Results from crystal violet staining, plate counts, and scanning electron microscopy (SEM) identified a single isolate from each of the O113, O145, O91, O157, and O121 serogroups that was capable of forming strong or moderate biofilms on stainless steel at 22 °C. However, the biofilm-forming strength of these five strains was reduced when incubation time progressed. Moreover, we found that these strains formed a dense pellicle at the air-liquid interface on stainless steel, which suggests that oxygen was conducive to biofilm formation. At 13 °C, biofilm formation by these strains decreased (P < 0.05), but gradually increased over time. Overall, STEC biofilm formation was most prominent at 22 °C up to 24 h. The findings in this study identify the environmental conditions that may promote STEC biofilm formation in food processing facilities and suggest that the ability of specific strains to form biofilms contributes to their persistence within these environments.

scholarly journals Biofilm formation by South African non-O157 Shiga toxigenic Escherichia coli on stainless steel coupons

2020 ◽  
Vol 66 (4) ◽  
pp. 328-336 ◽  
Author(s):  
Emmanuel W. Bumunang ◽  
Collins N. Ateba ◽  
Kim Stanford ◽  
Tim A. McAllister ◽  
Yan D. Niu
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
South African ◽  
Food Processing ◽  
Biofilm Formation ◽  
Viable Cell ◽  
Liquid Interface ◽  
Cell Counts ◽  
Potential Source ◽  
Air Liquid Interface

This study examined the biofilm-forming ability of six non-O157 Shiga-toxin-producing Escherichia coli (STEC) strains: O116:H21, wzx-Onovel5:H19, O129:H21, O129:H23, O26:H11, and O154:H10 on stainless steel coupons after 24, 48, and 72 h of incubation at 22 °C and after 168 h at 10 °C. The results of crystal violet staining revealed that strains O129:H23 and O154:H10 were able to form biofilms on both the submerged surface and the air–liquid interface of coupons, whereas strains O116:H21, wzx-Onovel5:H19, O129:H21, and O26:H11 formed biofilm only at the air–liquid interface. Viable cell counts and scanning electron microscopy showed that biofilm formation increased (p < 0.05) over time. The biofilm-forming ability of non-O157 STEC was strongest (p < 0.05) at 22 °C after 48 h of incubation. The strongest biofilm former regardless of temperature was O129:H23. Generally, at 10 °C, weak to no biofilm was observed for isolates O154:H10, O116:H21, wzx-Onovel5:H19, O26:H11, and O129:H21 after 168 h. This study found that temperature affected the biofilm-forming ability of non-O157 STEC strains. Overall, our data indicate a high potential for biofilm formation by the isolates at 22 °C, suggesting that non-O157 STEC strains could colonize stainless steel within food-processing facilities. This could serve as a potential source of adulteration and promote the dissemination of these potential pathogens in food.

Effects of Beef Juice on Biofilm Formation by Shiga Toxin–Producing Escherichia coli on Stainless Steel

2020 ◽  
Vol 17 (4) ◽  
pp. 235-242 ◽  
Author(s):  
Zhi Ma ◽  
Kim Stanford ◽  
Xiao M. Bie ◽  
Yan D. Niu ◽  
Tim A. McAllister
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Shiga Toxin ◽  
Biofilm Formation

scholarly journals Single- and Dual-Species Biofilm Formation by Shiga Toxin-Producing Escherichia coli and Salmonella, and Their Susceptibility to an Engineered Peptide WK2

2021 ◽  
Vol 9 (12) ◽  
pp. 2510
Author(s):  
Zhi Ma ◽  
Xia Tang ◽  
Kim Stanford ◽  
Xiaolong Chen ◽  
Tim A. McAllister ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Foodborne Pathogens ◽  
Shiga Toxin ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Single Species ◽  
Protein Labeling ◽  
Cross Contamination ◽  
Temporal And Spatial

Shiga toxin-producing Escherichia coli (STEC) and Salmonella enterica are important foodborne pathogens capable of forming both single- and multi-species biofilms. In this study, the mono- and dual-species biofilms were formed by STEC O113:H21 and Salmonella enterica serovar Choleraesuis 10708 on stainless steel in the presence of beef juice over 5 d at 22 °C. The dual-species biofilm mass was substantially (p < 0.05) greater than that produced by STEC O113:H21 or S. Choleraesuis 10708 alone. However, numbers (CFU/mL) of S. Choleraesuis 10708 or STEC O113:H21 cells in the dual-species biofilm were (p < 0.05) lower than their respective counts in single-species biofilms. In multi-species biofilms, the sensitivity of S. Choleraesuis 10708 to the antimicrobial peptide WK2 was reduced, but it was increased for STEC O113:H21. Visualization of the temporal and spatial development of dual-species biofilms using florescent protein labeling confirmed that WK2 reduced cell numbers within biofilms. Collectively, our results highlight the potential risk of cross-contamination by multi-species biofilms to food safety and suggest that WK2 may be developed as a novel antimicrobial or sanitizer for the control of biofilms on stainless steel.

scholarly journals Activity of Bacteriophage and Complex Tannins against Biofilm-Forming Shiga Toxin-Producing Escherichia coli from Canada and South Africa

Antibiotics ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 257
Author(s):  
Emmanuel W. Bumunang ◽  
Collins N. Ateba ◽  
Kim Stanford ◽  
Yan D. Niu ◽  
Y. Wang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
South Africa ◽  
Stainless Steel ◽  
Foodborne Pathogens ◽  
Shiga Toxin ◽  
Biofilm Formation ◽  
Condensed Tannin ◽  
Plant Secondary Metabolites ◽  
Brown Seaweed ◽  
E Coli

Bacteriophages, natural killers of bacteria, and plant secondary metabolites, such as condensed tannins, are potential agents for the control of foodborne pathogens. The first objective of this study evaluated the efficacy of a bacteriophage SA21RB in reducing pre-formed biofilms on stainless-steel produced by two Shiga toxin-producing Escherichia coli (STEC) strains, one from South Africa and the other from Canada. The second objective examined the anti-bacterial and anti-biofilm activity of condensed tannin (CT) from purple prairie clover and phlorotannins (PT) from brown seaweed against these strains. For 24-h-old biofilms, (O113:H21; 6.2 log10 colony-forming units per square centimeter (CFU/cm2) and O154:H10; 5.4 log10 CFU/cm2), 3 h of exposure to phage (1013 plaque-forming units per milliliter (PFU/mL)) reduced (p ≤ 0.05) the number of viable cells attached to stainless-steel coupons by 2.5 and 2.1 log10 CFU/cm2 for O113:H21 and O154:H10, respectively. However, as biofilms matured, the ability of phage to control biofilm formation declined. In biofilms formed for 72 h (O113:H21; 5.4 log10 CFU/cm2 and O154:H10; 7 log10 CFU/cm2), reductions after the same duration of phage treatment were only 0.9 and 1.3 log10 CFU/cm2 for O113:H21 and O154:H10, respectively. Initial screening of CT and PT for anti-bacterial activity by a microplate assay indicated that both STEC strains were less sensitive (p ≤ 0.05) to CT than PT over a concentration range of 25–400 µg/mL. Based on the lower activity of CT (25–400 µg/mL), they were not further examined. Accordingly, PT (50 µg/mL) inhibited (p ≤ 0.05) biofilm formation for up to 24 h of incubation at 22 °C, but this inhibition progressively declined over 72 h for both O154:H10 and O113:H21. Scanning electron microscopy revealed that both SA21RB and PT eliminated 24 h biofilms, but that both strains were able to adhere and form biofilms on stainless-steel coupons at longer incubation times. These findings revealed that phage SA21RB is more effective at disrupting 24 than 72 h biofilms and that PT were able to inhibit biofilm formation of both E. coli O154:H10 and O113:H21 for up to 24 h.

scholarly journals Application of Bacteriophages on Shiga Toxin-Producing Escherichia coli (STEC) Biofilm

Antibiotics ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1423
Author(s):  
Nicola Mangieri ◽  
Roberto Foschino ◽  
Claudia Picozzi
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Biofilm Formation ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Bacterial Cells ◽  
Abiotic Surfaces ◽  
Continuous Presence ◽  
Different Strains ◽  
Better Than

Shiga toxin-producing Escherichia coli are pathogenic bacteria able to form biofilms both on abiotic surfaces and on food, thus increasing risks for food consumers. Moreover, biofilms are difficult to remove and more resistant to antimicrobial agents compared to planktonic cells. Bacteriophages, natural predators of bacteria, can be used as an alternative to prevent biofilm formation or to remove pre-formed biofilm. In this work, four STEC able to produce biofilm were selected among 31 different strains and tested against single bacteriophages and two-phage cocktails. Results showed that our phages were able to reduce biofilm formation by 43.46% both when used as single phage preparation and as a cocktail formulation. Since one of the two cocktails had a slightly better performance, it was used to remove pre-existing biofilms. In this case, the phages were unable to destroy the biofilms and reduce the number of bacterial cells. Our data confirm that preventing biofilm formation in a food plant is better than trying to remove a preformed biofilm and the continuous presence of bacteriophages in the process environment could reduce the number of bacteria able to form biofilms and therefore improve the food safety.

Control of Listeria innocua Biofilms on Food Contact Surfaces with Slightly Acidic Electrolyzed Water and the Risk of Biofilm Cells Transfer to Duck Meat

2018 ◽  
Vol 81 (4) ◽  
pp. 582-592 ◽  
Author(s):  
HYE RI JEON ◽  
MI JIN KWON ◽  
KI SUN YOON
Keyword(s):  
Stainless Steel ◽  
Food Processing ◽  
Biofilm Formation ◽  
Listeria Innocua ◽  
Potential Hazard ◽  
Processing Efficiency ◽  
Electrolyzed Water ◽  
Food Contact ◽  
Food Contact Surfaces ◽  
Contact Surfaces

ABSTRACT Biofilm formation on food contact surfaces is a potential hazard leading to cross-contamination during food processing. We investigated Listeria innocua biofilm formation on various food contact surfaces and compared the washing effect of slightly acidic electrolyzed water (SAEW) at 30, 50, 70, and 120 ppm with that of 200 ppm of sodium hypochlorite (NaClO) on biofilm cells. The risk of L. innocua biofilm transfer and growth on food at retail markets was also investigated. The viability of biofilms that formed on food contact surfaces and then transferred cells to duck meat was confirmed by fluorescence microscopy. L. innocua biofilm formation was greatest on rubber, followed by polypropylene, glass, and stainless steel. Regardless of sanitizer type, washing removed biofilms from polypropylene and stainless steel better than from rubber and glass. Among the various SAEW concentrations, washing with 70 ppm of SAEW for 5 min significantly reduced L. innocua biofilms on food contact surfaces during food processing. Efficiency of transfer of L. innocua biofilm cells was the highest on polypropylene and lowest on stainless steel. The transferred biofilm cells grew to the maximum population density, and the lag time of transferred biofilm cells was longer than that of planktonic cells. The biofilm cells that transferred to duck meat coexisted with live, injured, and dead cells, which indicates that effective washing is essential to remove biofilm on food contact surfaces during food processing to reduce the risk of foodborne disease outbreaks.

Prevalence of Salmonella, Campylobacter, and Shiga Toxin–Producing Escherichia coli on the Surfaces of Raw Poultry Packages

2018 ◽  
Vol 81 (10) ◽  
pp. 1707-1712
Author(s):  
FUR-CHI CHEN ◽  
SANDRIA GODWIN ◽  
ANGELA GREEN ◽  
SHAHIDULLAH CHOWDHURY ◽  
RICHARD STONE
Keyword(s):  
Escherichia Coli ◽  
Foodborne Pathogens ◽  
Shiga Toxin ◽  
Microbial Contamination ◽  
Chicken Breast ◽  
Packaging Materials ◽  
Food Contact Surfaces ◽  
Plate Counts ◽  
Contact Surfaces ◽  
Raw Poultry

ABSTRACT Contamination on the exterior surfaces of raw poultry packages can be transmitted to hands and food contact surfaces during shopping and handling. This study compared the level of microbial contamination and prevalence of foodborne pathogens on the surfaces of raw poultry packages as related to the types of products, types of packaging, and packaging conditions. Packages of whole chicken, cut-up chicken (breast and leg quarter), and ground turkey were purchased from retail stores. Aerobic plate counts (APCs) were significantly different (P &lt; 0.05) among types of products and packaging materials, with ground turkey packages and the heat-sealed, high-walled containers being the lowest. APCs were significantly lower (P &lt; 0.05) when the packages were intact and tight compared with intact and loose. Of the 105 packages, there were 10 (9.5%) with the presence of either Shiga toxin–producing Escherichia coli (STEC) or Campylobacter; of those packages, 6 (5.7%) were positive for STEC, 7 (6.7%) were positive for Campylobacter, and 3 (2.9%) were positive for both pathogens on the surfaces. Salmonella was not detected on the surfaces of all tested packages. Surfaces of whole chicken packages were significantly (P &lt; 0.001) more likely to have detectable levels of Campylobacter and STEC than those of cut-up chicken packages. Packages that were positive for Campylobacter and/or STEC had significantly (P &lt; 0.005) higher APCs than negative packages. The results suggested that STEC is another significant pathogen present on the surfaces of poultry packages in addition to Campylobacter. The presence of STEC on the external packaging of raw poultry raises a concern because consumers may not expect such pathogens on the surfaces of poultry packages.

Control of the Biofilms Formed by Curli- and Cellulose-Expressing Shiga Toxin–Producing Escherichia coli Using Treatments with Organic Acids and Commercial Sanitizers

2015 ◽  
Vol 78 (5) ◽  
pp. 990-995 ◽  
Author(s):  
YOEN JU PARK ◽  
JINRU CHEN
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Organic Acids ◽  
Shiga Toxin ◽  
Cell Contact ◽  
Bacterial Cells ◽  
Bacterial Surface ◽  
Additional Information ◽  
Steel Surfaces ◽  
Biological Protection

Biofilms are a mixture of bacteria and extracellular products secreted by bacterial cells and are of great concern to the food industry because they offer physical, mechanical, and biological protection to bacterial cells. This study was conducted to quantify biofilms formed by different Shiga toxin–producing Escherichia coli (STEC) strains on polystyrene and stainless steel surfaces and to determine the effectiveness of sanitizing treatments in control of these biofilms. STEC producing various amounts of cellulose (n = 6) or curli (n = 6) were allowed to develop biofilms on polystyrene and stainless steel surfaces at 28°C for 7 days. The biofilms were treated with 2% acetic or lactic acid and manufacturer-recommended concentrations of acidic or alkaline sanitizers, and residual biofilms were quantified. Treatments with the acidic and alkaline sanitizers were more effective than those with the organic acids for removing the biofilms. Compared with their counterparts, cells expressing a greater amount of cellulose or curli formed more biofilm mass and had greater residual mass after sanitizing treatments on polystyrene than on stainless steel. Research suggests that the organic acids and sanitizers used in the present study differed in their ability to control biofilms. Bacterial surface components and cell contact surfaces can influence both biofilm formation and the efficacy of sanitizing treatments. These results provide additional information on control of biofilms formed by STEC.

Growth of Listeria monocytogenes as a Biofilm on Various Food-Processing Surfaces

1996 ◽  
Vol 59 (8) ◽  
pp. 827-831 ◽  
Author(s):  
ISABEL C. BLACKMAN ◽  
JOSEPH F. FRANK
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Food Processing ◽  
Biofilm Formation ◽  
Minimal Medium ◽  
Processing Plant ◽  
Biofilm Growth ◽  
Sanitary Condition ◽  
Plant Environment ◽  
Substantial Risk

The objective of this research was to determine the ability of Listeria monocytogenes to grow as a biofilm on various food-processing surfaces including stainless steel, Teflon®, nylon, and polyester floor sealant. Each of these surfaces was able to support biofilm formation when incubation was at 21°C in Trypticase soy broth (TSB). Biofilm formation was greatest on polyester floor sealant (40% of surface area covered after 7 days of incubation) and least on nylon (3% coverage). The use of chemically defined minimal medium resulted in a lack of biofilm formation on polyester floor sealant, and reduced biofilm levels on stainless steel. Biofilm formation was reduced with incubation at 10°C, but Teflon® and stainless steel still allowed 23 to 24% coverage after incubation in TSB for 18 days. Biofilm growth of L. monocytogenes was sufficient to provide a substantial risk of this pathogen contaminating the food-processing plant environment if wet surfaces are not maintained in a sanitary condition.

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