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 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 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 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.

scholarly journals Biofilm Formation by Escherichia coli O157:H7 on Stainless Steel: Effect of Exopolysaccharide and Curli Production on Its Resistance to Chlorine

2005 ◽  
Vol 71 (1) ◽  
pp. 247-254 ◽  
Author(s):  
Jee-Hoon Ryu ◽  
Larry R. Beuchat
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Biofilm Formation ◽  
Escherichia Coli O157 ◽  
Strain Atcc ◽  
Planktonic Cells ◽  
E Coli ◽  
Biofilm Production ◽  
Resistant Population ◽  
Population Sizes

ABSTRACT The resistance of Escherichia coli O157:H7 strains ATCC 43895-, 43895-EPS (an exopolysaccharide [EPS]-overproducing mutant), and ATCC 43895+ (a curli-producing mutant) to chlorine, a sanitizer commonly used in the food industry, was studied. Planktonic cells of strains 43895-EPS and/or ATCC 43895+ grown under conditions supporting EPS and curli production, respectively, showed the highest resistance to chlorine, indicating that EPS and curli afford protection. Planktonic cells (ca. 9 log10 CFU/ml) of all strains, however, were killed within 10 min by treatment with 50 μg of chlorine/ml. Significantly lower numbers of strain 43895-EPS, compared to those of strain ATCC 43895-, attached to stainless steel coupons, but the growth rate of strain 43895-EPS on coupons was not significantly different from that of strain ATCC 43895-, indicating that EPS production did not affect cell growth during biofilm formation. Curli production did not affect the initial attachment of cells to coupons but did enhance biofilm production. The resistance of E. coli O157:H7 to chlorine increased significantly as cells formed biofilm on coupons; strain ATCC 43895+ was the most resistant. Population sizes of strains ATCC 43895+ and ATCC 43895- in biofilm formed at 12�C were not significantly different, but cells of strain ATCC 43895+ showed significantly higher resistance than did cells of strain ATCC 43895-. These observations support the hypothesis that the production of EPS and curli increase the resistance of E. coli O157:H7 to chlorine.

scholarly journals Mixed Biofilm Formation by Shiga Toxin–Producing Escherichia coli and Salmonella enterica Serovar Typhimurium Enhanced Bacterial Resistance to Sanitization due to Extracellular Polymeric Substances†

2013 ◽  
Vol 76 (9) ◽  
pp. 1513-1522 ◽  
Author(s):  
RONG WANG ◽  
NORASAK KALCHAYANAND ◽  
JOHN W. SCHMIDT ◽  
DAYNA M. HARHAY
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Foodborne Pathogens ◽  
Shiga Toxin ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Bacterial Resistance ◽  
Salmonella Enterica Serovar Typhimurium ◽  
E Coli ◽  
Serovar Typhimurium

Shiga toxin–producing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium are important foodborne pathogens capable of forming single-species biofilms or coexisting in multispecies biofilm communities. Bacterial biofilm cells are usually more resistant to sanitization than their planktonic counterparts, so these foodborne pathogens in biofilms pose a serious food safety concern. We investigated how the coexistence of E. coli O157:H7 and Salmonella Typhimurium strains would affect bacterial planktonic growth competition and mixed biofilm composition. Furthermore, we also investigated how mixed biofilm formation would affect bacterial resistance to common sanitizers. Salmonella Typhimurium strains were able to outcompete E. coli strains in the planktonic growth phase; however, mixed biofilm development was highly dependent upon companion strain properties in terms of the expression of bacterial extracellular polymeric substances (EPS), including curli fimbriae and exopolysaccharide cellulose. The EPS-producing strains with higher biofilm-forming abilities were able to establish themselves in mixed biofilms more efficiently. In comparison to single-strain biofilms, Salmonella or E. coli strains with negative EPS expression obtained significantly enhanced resistance to sanitization by forming mixed biofilms with an EPS-producing companion strain of the other species. These observations indicate that the bacterial EPS components not only enhance the sanitizer resistance of the EPS-producing strains but also render protections to their companion strains, regardless of species, in mixed biofilms. Our study highlights the potential risk of cross-contamination by multispecies biofilms in food safety and the need for increased attention to proper sanitization practices in food processing facilities.

scholarly journals Role of bolA and rpoS genes in biofilm formation and adherence pattern by Escherichia coli K-12 MG1655 on polypropylene, stainless steel, and silicone surfaces

2017 ◽  
Vol 64 (2) ◽  
pp. 179-189 ◽  
Author(s):  
Mohd Adnan ◽  
Ana Margarida Sousa ◽  
Idalina Machado ◽  
Maria Olivia Pereira ◽  
Saif Khan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Biofilm Formation ◽  
Adherence Pattern ◽  
K 12

scholarly journals Biofilm Formation Potential of Heat-Resistant Escherichia coli Dairy Isolates and the Complete Genome of Multidrug-Resistant, Heat-Resistant Strain FAM21845

2017 ◽  
Vol 83 (15) ◽  
Author(s):  
Roger Marti ◽  
Michael Schmid ◽  
Sandra Kulli ◽  
Kerstin Schneeberger ◽  
Javorka Naskova ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Biofilm Formation ◽  
Food Production ◽  
Osmotic Shock ◽  
Multidrug Resistant ◽  
Formation Potential ◽  
Conjugative Plasmids ◽  
Heat Resistant ◽  
Initial Adhesion

ABSTRACT We tested the biofilm formation potential of 30 heat-resistant and 6 heat-sensitive Escherichia coli dairy isolates. Production of curli and cellulose, static biofilm formation on polystyrene (PS) and stainless steel surfaces, biofilm formation under dynamic conditions (Bioflux), and initial adhesion rates (IAR) were evaluated. Biofilm formation varied greatly between strains, media, and assays. Our results highlight the importance of the experimental setup in determining biofilm formation under conditions of interest, as correlation between different assays was often not a given. The heat-resistant, multidrug-resistant (MDR) strain FAM21845 showed the strongest biofilm formation on PS and the highest IAR and was the only strain that formed significant biofilms on stainless steel under conditions relevant to the dairy industry, and it was therefore fully sequenced. Its chromosome is 4.9 Mb long, and it harbors a total of five plasmids (147.2, 54.2, 5.8, 2.5, and 1.9 kb). The strain carries a broad range of genes relevant to antimicrobial resistance and biofilm formation, including some on its two large conjugative plasmids, as demonstrated in plate mating assays. IMPORTANCE In biofilms, cells are embedded in an extracellular matrix that protects them from stresses, such as UV radiation, osmotic shock, desiccation, antibiotics, and predation. Biofilm formation is a major bacterial persistence factor of great concern in the clinic and the food industry. Many tested strains formed strong biofilms, and especially strains such as the heat-resistant, MDR strain FAM21845 may pose a serious issue for food production. Strong biofilm formation combined with diverse resistances (some encoded on conjugative plasmids) may allow for increased persistence, coselection, and possible transfer of these resistance factors. Horizontal gene transfer may conceivably occur in the food production setting or the gastrointestinal tract after consumption.

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