scholarly journals Efficacy of Novel Bacteriophages against Escherichia coli Biofilms on Stainless Steel

Antibiotics ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1150
Author(s):  
Jean Pierre González-Gómez ◽  
Berenice González-Torres ◽  
Pedro Javier Guerrero-Medina ◽  
Osvaldo López-Cuevas ◽  
Cristóbal Chaidez ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Development Stage ◽  
Lytic Activity ◽  
Meat Industry ◽  
Promising Alternative ◽  
E Coli ◽  
Adhesion Ability ◽  
Biofilm Development

Biofilm formation by E. coli is a serious threat to meat processing plants. Chemical disinfectants often fail to eliminate biofilms; thus, bacteriophages are a promising alternative to solve this problem, since they are widely distributed, environmentally friendly, and nontoxic to humans. In this study, the biofilm formation of 10 E. coli strains isolated from the meat industry and E. coli ATCC BAA-1430 and ATCC 11303 were evaluated. Three strains, isolated from the meat contact surfaces, showed adhesion ability and produced extracellular polymeric substances. Biofilms of these three strains were developed onto stainless steel (SS) surfaces and enumerated at 2, 12, 24, 48, and 120 h, and were visualized by scanning electron microscopy. Subsequently, three bacteriophages showing podovirus morphology were isolated from ground beef and poultry liver samples, which showed lytic activity against the abovementioned biofilm-forming strains. SS surfaces with biofilms of 2, 14, and 48 h maturity were treated with mixed and individual bacteriophages at 8 and 9 log10 PFU/mL for 1 h. The results showed reductions greater than 6 log10 CFU/cm2 as a result of exposing SS surfaces with biofilms of 24 h maturity to 9 log10 PFU/mL of bacteriophages; however, the E. coli and bacteriophage strains, phage concentration, and biofilm development stage had significant effects on biofilm reduction (p < 0.05). In conclusion, the isolated bacteriophages showed effectiveness at reducing biofilms of isolated E. coli; however, it is necessary to increase the libraries of phages with lytic activity against the strains isolated from production environments.

scholarly journals Innovative Control of Biofilms on Stainless Steel Surfaces Using Electrolyzed Water in the Dairy Industry

Foods ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 103
Author(s):  
Rodrigo Jiménez-Pichardo ◽  
Iriana Hernández-Martínez ◽  
Carlos Regalado-González ◽  
José Santos-Cruz ◽  
Yunny Meas-Vong ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Biofilm Formation ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Dairy Industry ◽  
Raw Milk ◽  
Steel Plates ◽  
Heat Processing ◽  
Rrna Gene ◽  
Electrolyzed Water ◽  
Biofilm Development

Biofilms on food-contact surfaces can lead to recurrent contamination. This work aimed to study the biofilm formation process on stainless steel plates used in the dairy industry: 304 surface finish 2B and electropolished; and the effect of a cleaning and disinfection process using alkaline (AEW) and neutral (NEW) electrolyzed water. Milk fouling during heat processing can lead to type A or B deposits, which were analyzed for composition, surface energy, thickness, and roughness, while the role of raw milk microbiota on biofilm development was investigated. Bacteria, yeasts, and lactic acid bacteria were detected using EUB-338, PF2, and Str-493 probes, respectively, whereas Lis-637 probe detected Listeria sp. The genetic complexity and diversity of biofilms varied according to biofilm maturation day, as evaluated by 16S rRNA gene sequence, denaturing gradient gel electrophoresis, and fluorescence in situ hybridization microscopy. From analysis of the experimental designs, a cleaning stage of 50 mg/L NaOH of AEW at 30 °C for 10 min, followed by disinfection using 50 mg/L total available chlorine of NEW at 20 °C for 5 min is a sustainable alternative process to prevent biofilm formation. Fluorescence microscopy was used to visualize the effectiveness of this process.

scholarly journals Interaction of Salmonella with E. coli and Proteus spp. in Biofilm Formation

2021 ◽  
pp. 30-45
Author(s):  
S. U. Pathiranage ◽  
D. N. N. Madushanka ◽  
K. V. D. M. Hasintha ◽  
H. C. Nadishani ◽  
G. C. P. Fernando ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Broiler Chicken ◽  
Chicken Meat ◽  
Salmonella Spp ◽  
E Coli ◽  
Time Points ◽  
Dual Cultures ◽  
Microtitre Plate Assay ◽  
Biofilm Development ◽  
Numerous Time

Aims: Investigate the interaction of Salmonella spp. with E. coli and Proteus spp. in biofilm formation as mono and dual-species at different time durations Experimental Design: Salmonella, Proteus, and E. coli were isolated from Broiler chicken meat, and the biofilm-forming ability of these organisms were studied. Place and Duration of Study: The study was conducted at the Laboratory of Livestock Production, Faculty of Agricultural Sciences, Sabaragamuwa University of Sri Lanka, from 2019 December to 2020 May. Methodology: This study investigated the biofilm-forming ability of Salmonella as a mono species and its interaction with E. coli and Proteus in the process of biofilm formation. Microorganisms used for this study were isolated from broiler chicken meat. Biofilm was quantified using a microtitre plate assay. The interaction effects were tested at the temperature of 280C in different time durations (up to 120 hours). Results: Salmonella 1 and Proteus monocultures showed significantly higher biofilm-forming ability than Salmonella 3 isolate at all tested time points. At 120 hr, additionally to the salmonella 1 and Proteus isolates E. coli also formed significantly higher biofilms than Salmonella 3. However, Salmonella 3 was the lowest biofilm former as mono biofilm at all tested time durations. Salmonella 1 interaction with Salmonella 3 isolates formed less biofilms than Salmonella 1 mono biofilm at 48hr and 72hr correspondingly. Salmonella 1 and its interactions with Salmonella 3, Proteus, E. coli showed similar biofilm-forming abilities without significant differences at all other tested time points. Specifically, Salmonella 3 interaction with Salmonella 1 as dual biofilm showed higher biofilm-forming ability than Salmonella 3 mono biofilm at all tested time points. Tested isolates and their interaction achieved the highest biofilm formation at numerous time points. In fact, at 48hr, Salmonella 3 isolates and its interaction of Proteus, E. coli, and Salmonella 1 interaction with Proteus attained their highest biofilm formation abilities. The highest biofilm formation was achieved by Salmonella 1 isolate as mono biofilm and Salmonella 1 interaction with E. coli as dual biofilm at 72hr. Biofilm-forming trend of respective isolates and interactions showed numerous patterns at tested time durations. Specifically, E. coli rapidly enhanced its biofilm-forming ability as monoculture from 24 hr to 120 hr. Proteus, Salmonella 3 as monocultures, Salmonella 3 interaction with Proteus and E. coli as dual cultures showed progressive biofilm development from 24 hr to 48 hr. Salmonella 1 monoculture and its interaction with Salmonella 3, E. coli as dual biofilm improved their biofilm-forming ability from 24 hr to 72 hr. Similar to Salmonella 3 interaction with Proteus, Salmonella 1 interaction with Proteus also increased its biofilm-forming ability from 24 hr to 48 hr. Conclusions: This study concluded that there is a variation among isolates and their combinations in forming the biofilms, where there is an enhancement of biofilm in dual-species over the mono-species in some interaction, and there is a reduction in biofilm formation by dual-species with some combinations. Further, this concluded that Salmonella is interacting with other commonly found bacteria such as Proteus and E. coli in biofilm formation.

scholarly journals Antimicrobial Peptides-Coated Stainless Steel for Fighting Biofilms Formation for Food and Medical Fields: Review of Literature

Coatings ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1216
Author(s):  
Mayssane Hage ◽  
Hikmat Akoum ◽  
Nour-Eddine Chihib ◽  
Charafeddine Jama
Keyword(s):  
Stainless Steel ◽  
Antimicrobial Peptides ◽  
Bacterial Adhesion ◽  
Biofilm Formation ◽  
Bacterial Biofilms ◽  
Food Environments ◽  
Review Of Literature ◽  
Safe Alternative ◽  
Antimicrobial Coatings ◽  
Biofilm Development

Emerging technology regarding antimicrobial coatings contributes to fighting the challenge of pathogenic bacterial biofilms in medical and agri-food environments. Stainless steel is a material widely used in those fields since it has satisfying mechanical properties, but it, unfortunately, lacks the required bio-functionality, rendering it vulnerable to bacterial adhesion and biofilm formation. Therefore, this review aims to present the coatings developed by employing biocides grafted on stainless steel. It also highlights antimicrobial peptides (AMPs)used to coat stainless steel, particularly nisin, which is commonly accepted as a safe alternative to prevent pathogenic biofilm development.

Cyclic Changes in the Amide Bands Within Escherichia coli Biofilms Monitored Using Real-Time Infrared Attenuated Total Reflection Spectroscopy (IR-ATR)

2019 ◽  
Vol 73 (4) ◽  
pp. 424-432 ◽  
Author(s):  
Pavla Stenclova ◽  
Simon Freisinger ◽  
Holger Barth ◽  
Alexander Kromka ◽  
Boris Mizaikoff
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Total Reflection ◽  
Attenuated Total Reflection ◽  
Continuous Increase ◽  
Spectral Window ◽  
E Coli ◽  
Cyclic Changes

Contrary to the planktonic state of bacteria, their biofilm form represents severe complications in areas such as human medicine or food industry due to the increasing resistance against harsh conditions and treatment. In the present study, infrared attenuated total reflection (IR-ATR) spectroscopy has been applied as an analytic tool studying Escherichia coli ( E. coli) biofilm formation close to real time. We report on IR spectroscopic investigations on the biofilm formation via ATR waveguides probing the biofilm in the spectral window of 1800–900 cm−1 at dynamic flow conditions, which facilitated monitoring the growth dynamics during several days. Key IR bands are in the range 1700–1590 cm−1 (amide I), 1580–1490 cm−1 (amide II), and 1141–1006 cm−1 extracellular polymeric substances (EPS), which were evaluated as a function of time. Cyclic fluctuations of the amide I and amide II bands and a continuous increase of the EPS band were related to the starvation of bottom-layered bacteria caused by the nutrient gradient. Potential death of bacteria may then result in cannibalistic behavior known for E. coli colonies. Observing this behavior via IR spectroscopy allows revealing these cyclical changes in bottom-layered bacteria within the biofilm under continuous nutrient flow, in molecular detail, and during extended periods for the first time.

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.

Biofilm Development and Sanitizer Inactivation of Listeria monocytogenes and Salmonella typhimurium on Stainless Steel and Buna-n Rubber

1993 ◽  
Vol 56 (9) ◽  
pp. 750-758 ◽  
Author(s):  
AMY B. RONNER ◽  
AMY C. L. WONG
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Biofilm Formation ◽  
Bacterial Biofilm ◽  
Extracellular Matrices ◽  
Nutrient Level ◽  
Extracellular Materials ◽  
Biofilm Development ◽  
Nutrient Conditions

Biofilm formation by seven strains of Listeria monocytogenes and one strain of Salmonella typhimurium on stainless steel and Buna-n rubber was examined under two nutrient conditions. The type of surface, nutrient level, and organism influenced biofilm development and production of extracellular materials. Buna-n had a strong bacteriostatic effect on L. monocytogenes, and biofilm formation on Buna-n under low nutrient conditions was reduced for four of the seven strains tested. Buna-n was less bacteriostatic toward S. typhimurium. It inhibited the growth of several other pathogens to varying degrees. An ethylene propylene diamine monomer rubber was less inhibitory than Buna-n, and Viton rubber had no effect. The effectiveness of sanitizers on biofilm bacteria was examined. Biofilms were challenged with four types of detergent and nondetergent sanitizers. Resistance to sanitizers was strongly influenced by the type of surface. Bacterial biofilm populations on stainless steel were reduced 3–5 log by all the sanitizers, but those on Buna-n were resistant to these sanitizers and were reduced less than 1–2 log. In contrast, planktonic (suspended) bacteria were reduced 7–8 log by these sanitizers. Chlorine and anionic acid sanitizers generally removed extracellular materials from biofilms better than iodine and quaternary ammonium detergent sanitizers. Scanning electron microscopy demonstrated that biofilm cells and extracellular matrices could remain on sanitized biofilm cells and extracellular matrices could remain surfaces from which no viable cells were recovered.

scholarly journals Bacteriophages as an Alternative Strategy for Fighting Biofilm Development

2014 ◽  
Vol 63 (2) ◽  
pp. 137-145 ◽  
Author(s):  
SYLWIA PARASION ◽  
MAGDALENA KWIATEK ◽  
ROMUALD GRYKO ◽  
LIDIA MIZAK ◽  
ANNA MALM
Keyword(s):  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Bacterial Resistance ◽  
Bacterial Species ◽  
Experimental Studies ◽  
Alternative Methods ◽  
Resistant Bacteria ◽  
Bacterial Cells ◽  
Coagulase Negative Staphylococci ◽  
Biofilm Development

The ability of microbes to form biofilms is an important element of their pathogenicity, and biofilm formation is a serious challenge for today's medicine. Fighting the clinical complications associated with biofilm formation is very difficult and linked to a high risk of failure, especially in a time of increasing bacterial resistance to antibiotics. Bacterial species most commonly isolated from biofilms include coagulase-negative staphylococci, Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter spp. The frequent failure of antibiotic therapy led researchers to look for alternative methods and experiment with the use of antibacterial factors with a mechanism of action different from that of antibiotics. Experimental studies with bacteriophages and mixtures thereof, expressing lytic properties against numerous biofilm-forming bacterial species showed that bacteriophages may both prevent biofilm formation and contribute to eradication of biofilm bacteria. A specific role is played here by phage depolymerases, which facilitate the degradation of extracellular polymeric substances (EPS) and thus the permeation of bacteriophages into deeper biofilm layers and lysis of the susceptible bacterial cells. Much hope is placed in genetic modifications of bacteriophages that would allow the equipping bacteriophages with the function of depolymerase synthesis. The use of phage cocktails prevents the development of phage-resistant bacteria.

scholarly journals Biofilm Formation and Dispersal under the Influence of the Global Regulator CsrA of Escherichia coli

2002 ◽  
Vol 184 (1) ◽  
pp. 290-301 ◽  
Author(s):  
Debra W. Jackson ◽  
Kazushi Suzuki ◽  
Lawrence Oakford ◽  
Jerry W. Simecka ◽  
Mark E. Hart ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Rna Binding ◽  
Glycogen Synthase ◽  
Regulatory Protein ◽  
Ectopic Expression ◽  
Knockout Mutant ◽  
E Coli ◽  
K 12 ◽  
Biofilm Development

ABSTRACT The predominant mode of growth of bacteria in the environment is within sessile, matrix-enclosed communities known as biofilms. Biofilms often complicate chronic and difficult-to-treat infections by protecting bacteria from the immune system, decreasing antibiotic efficacy, and dispersing planktonic cells to distant body sites. While the biology of bacterial biofilms has become a major focus of microbial research, the regulatory mechanisms of biofilm development remain poorly defined and those of dispersal are unknown. Here we establish that the RNA binding global regulatory protein CsrA (carbon storage regulator) of Escherichia coli K-12 serves as both a repressor of biofilm formation and an activator of biofilm dispersal under a variety of culture conditions. Ectopic expression of the E. coli K-12 csrA gene repressed biofilm formation by related bacterial pathogens. A csrA knockout mutation enhanced biofilm formation in E. coli strains that were defective for extracellular, surface, or regulatory factors previously implicated in biofilm formation. In contrast, this csrA mutation did not affect biofilm formation by a glgA (glycogen synthase) knockout mutant. Complementation studies with glg genes provided further genetic evidence that the effects of CsrA on biofilm formation are mediated largely through the regulation of intracellular glycogen biosynthesis and catabolism. Finally, the expression of a chromosomally encoded csrA′-′lacZ translational fusion was dynamically regulated during biofilm formation in a pattern consistent with its role as a repressor. We propose that global regulation of central carbon flux by CsrA is an extremely important feature of E. coli biofilm development.

Attachment and Biofilm Formation by Escherichia coli O157:H7 on Stainless Steel as Influenced by Exopolysaccharide Production, Nutrient Availability, and Temperature

2004 ◽  
Vol 67 (10) ◽  
pp. 2123-2131 ◽  
Author(s):  
JEE-HOON RYU ◽  
HOIKYUNG KIM ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Stainless Steel ◽  
Nutrient Availability ◽  
Biofilm Formation ◽  
Escherichia Coli O157 ◽  
Strain Atcc ◽  
Wild Type ◽  
E Coli ◽  
Nutrient Environment ◽  
Phosphate Buffered Saline

The influence of exopolysaccharide (EPS) production, nutrient availability, and temperature on attachment and biofilm formation by Escherichia coli O157:H7 strains ATCC 43895 (wild type) and 43895-EPS (extensive EPS-producing mutant) on stainless steel coupons (SSCs) was investigated. Cells grown on heated lettuce juice agar and modified tryptic soy agar were suspended in phosphate-buffered saline (PBS). SSCs were immersed in the cell suspension (109 CFU/ml) at 4°C for 24 h. Biofilm formation by cells attached to SSCs as affected by immersing in 10% tryptic soy broth (TSB), lettuce juice broth (LJB), and minimal salts broth (MSB) at 12 and 22°C was studied. A significantly lower number of strain 43895-EPS cells, compared to strain ATCC 43895 cells, attached to SSCs during a 24-h incubation (4°C) period in PBS suspension. Neither strain formed a biofilm on SSCs subsequently immersed in 10% TSB or LJB, but both strains formed biofilms in MSB. Populations of attached cells and planktonic cells of strain ATCC 43895 gradually decreased during incubation for 6 days in LJB at 22°C, but populations of strain 43895-EPS remained constant for 6 days at 22°C, indicating that the EPS-producing mutant, compared to the wild-type strain, has a higher tolerance to the low-nutrient environment presented by LJB. It is concluded that EPS production by E. coli O157:H7 inhibits attachment to SSCs and that reduced nutrient availability enhances biofilm formation. Biofilms formed under conditions favorable for EPS production may protect E. coli O157:H7 against sanitizers used to decontaminate lettuce and produce processing environments. Studies are under way to test this hypothesis.

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