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.

scholarly journals Biofilm Formation by Listeria monocytogenes 15G01, a Persistent Isolate from a Seafood-Processing Plant, Is Influenced by Inactivation of Multiple Genes Belonging to Different Functional Groups

2021 ◽  
Vol 87 (10) ◽  
Author(s):  
Jessika Nowak ◽  
Sandra B. Visnovsky ◽  
Andrew R. Pitman ◽  
Cristina D. Cruz ◽  
Jon Palmer ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Biofilm Formation ◽  
Processing Plant ◽  
Wild Type ◽  
Content Type ◽  
Multiple Genes ◽  
Processing Plants ◽  
Biofilm Structure ◽  
Seafood Processing

ABSTRACT Listeria monocytogenes is a ubiquitous foodborne pathogen that results in a high rate of mortality in sensitive and immunocompromised people. Contamination of food with L. monocytogenes is thought to occur during food processing, most often as a result of the pathogen producing a biofilm that persists in the environment and acting as the source for subsequent dispersal of cells onto food. A survey of seafood-processing plants in New Zealand identified the persistent strain 15G01, which has a high capacity to form biofilms. In this study, a transposon library of L. monocytogenes 15G01 was screened for mutants with altered biofilm formation, assessed by a crystal violet assay, to identify genes involved in biofilm formation. This screen identified 36 transposants that showed a significant change in biofilm formation compared to the wild type. The insertion sites were in 27 genes, 20 of which led to decreased biofilm formation and seven to an increase. Two insertions were in intergenic regions. Annotation of the genes suggested that they are involved in diverse cellular processes, including stress response, autolysis, transporter systems, and cell wall/membrane synthesis. Analysis of the biofilms produced by the transposants using scanning electron microscopy and fluorescence microscopy showed notable differences in the structure of the biofilms compared to the wild type. In particular, inactivation of uvrB and mltD produced coccoid-shaped cells and elongated cells in long chains, respectively, and the mgtB mutant produced a unique biofilm with a sandwich structure which was reversed to the wild-type level upon magnesium addition. The mltD transposant was successfully complemented with the wild-type gene, whereas the phenotypes were not or only partially restored for the remaining mutants. IMPORTANCE The major source of contamination of food with Listeria monocytogenes is thought to be due to biofilm formation and/or persistence in food-processing plants. By establishing as a biofilm, L. monocytogenes cells become harder to eradicate due to their increased resistance to environmental threats. Understanding the genes involved in biofilm formation and their influence on biofilm structure will help identify new ways to eliminate harmful biofilms in food processing environments. To date, multiple genes have been identified as being involved in biofilm formation by L. monocytogenes; however, the exact mechanism remains unclear. This study identified four genes associated with biofilm formation by a persistent strain. Extensive microscopic analysis illustrated the effect of the disruption of mgtB, clsA, uvrB, and mltD and the influence of magnesium on the biofilm structure. The results strongly suggest an involvement in biofilm formation for the four genes and provide a basis for further studies to analyze gene regulation to assess the specific role of these biofilm-associated genes.

scholarly journals Antibiofilm Effects of Nanoparticles and Visible Light Illumination Against Listeria monocytogenes

2021 ◽  
Vol 12 ◽  
Author(s):  
Sanna Puranen ◽  
Kati Riekkinen ◽  
Jenni Korhonen
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Food Safety ◽  
Visible Light ◽  
Food Processing ◽  
Biofilm Formation ◽  
Plate Count ◽  
Light Illumination ◽  
Visible Light Illumination ◽  
Aluminum Plates

Listeria monocytogenes bacteria pose a particular risk to the food industry as the species is known to form biofilm and to survive in a wide range of challenging environmental conditions. L. monocytogenes can cause listeriosis, a serious food-borne disease, and effective and safe antibiofilm materials and sanitary methods for food processing environments are intensively sought. A variety of nanoparticle materials have been recognized as safe to use in food environments, which allows the application of nanomaterials also for food safety purposes. Nanoparticles together with light illumination generate reactive oxygen species which inactivate bacteria by breaking down cell membranes, proteins, and DNA. The main objective of this study was to evaluate the efficacy of nanomaterials and blue light illumination for L. monocytogenes ATCC 7644 biofilm inactivation. Biofilm was allowed to form for 72 h on nanocoated stainless steel and aluminum plates, after which the plates were illuminated. Non-coated control plates were used to evaluate the antibiofilm efficacy of nanocoating. Plate count method was used to evaluate bacteria counts after illumination. Nanocoating did not affect initial biofilm formation compared to the control plates. Biofilm was significantly (p < 0.05) reduced on stainless steel, aluminum, and TiO2-coated aluminum plates after 72-h illumination by 1.9, 3.2, and 5.9 log, respectively. Nanocoating with visible light illumination could be an effective and safe method for enhancing food safety in food processing facilities to control biofilm formation. Evidence of antibiofilm properties of nanomaterials together with visible light illumination is limited; hence, future studies with variable light intensities and nanomaterials are needed.

scholarly journals Control of Growth and Persistence of Listeria monocytogenes and β-Lactam-Resistant Escherichia coli by Thymol in Food Processing Settings

Molecules ◽  
2020 ◽  
Vol 25 (2) ◽  
pp. 383 ◽  
Author(s):  
Maria Grazia Cusimano ◽  
Vita Di Stefano ◽  
Maria La Giglia ◽  
Vincenzo Di Marco Lo Presti ◽  
Domenico Schillaci ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Food Processing ◽  
Biofilm Formation ◽  
Bacterial Biofilm ◽  
Free Living ◽  
Biofilm Growth ◽  
Food Industries ◽  
E Coli ◽  
Reference Strains

The main objective of this study was to evaluate the efficacy of thymol in controlling environmental contamination in food processing facilities. The effect of thymol was tested as an agent to prevent planktonic and bacterial biofilm growth of twenty-five Listeria monocytogenes isolates from a variety of foods and five Escherichia coli isolates from a farm. The E. coli isolates were positive for extended spectrum β-lactamase (ESBL) genes. All isolates and reference strains were susceptible to thymol at Minimum inhibitory concentration (MIC) values ranging from 250 to 800 μg/mL. An interesting activity of interference with biofilm formation of L. monocytogenes and E. coli was found for thymol at sub-MIC concentrations of 200, 100, 75, and 50 μg/mL. Anti-biofilm activity ranging from 59.71% to 66.90% against pre-formed 24-h-old L. monocytogenes biofilms at concentrations of 500 or 800 µg/mL, corresponding to 2× MIC, was determined against free-living forms of six isolates chosen as the best or moderate biofilm producers among the tested strains. The property of thymol to attack L. monocytogenes biofilm formation was also observed at a concentration of 100 µg/mL, corresponding to 1/4 MIC, by using a stainless-steel model to simulate the surfaces in food industries. This study gives information on the use of thymol in food processing setting.

Morphological Change and Decreasing Transfer Rate of Biofilm-Featured Listeria monocytogenes EGDe

2017 ◽  
Vol 80 (3) ◽  
pp. 368-375 ◽  
Author(s):  
Yuejia Lee ◽  
Chinling Wang
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Food Processing ◽  
Biofilm Formation ◽  
Transfer Rate ◽  
Initial Level ◽  
Foodborne Pathogen ◽  
Bacterial Growth Rate ◽  
Biofilm Development ◽  
Food Processing Environment

ABSTRACT Listeria monocytogenes, a lethal foodborne pathogen, has the ability to resist the hostile food processing environment and thus frequently contaminates ready-to-eat foods during processing. It is commonly accepted that the tendency of L. monocytogenes' to generate biofilms on various surfaces enhances its resistance to the harshness of the food processing environment. However, the role of biofilm formation in the transferability of L. monocytogenes EGDe remains controversial. We examined the growth of Listeria biofilms on stainless steel surfaces and their effect on the transferability of L. monocytogenes EGDe. The experiments were a factorial 2 × 2 design with at least three biological replicates. Through scanning electron microscopy, a mature biofilm with intensive aggregates of cells was observed on the surface of stainless steel after 3 or 5 days of incubation, depending on the initial level of inoculation. During biofilm development, L. monocytogenes EGDe carried out binary fission vigorously before a mature biofilm was formed and subsequently changed its cellular morphology from rod shaped to sphere shaped. Furthermore, static biofilm, which was formed after 3 days of incubation at 25°C, significantly inhibited the transfer rate of L. monocytogenes EGDe from stainless steel blades to 15 bologna slices. During 7 days of storage at 4°C, however, bacterial growth rate was not significantly impacted by whether bacteria were transferred from biofilm and the initial concentrations of transferred bacteria on the slice. In conclusion, this study is the first to report a distinct change in morphology of L. monocytogenes EGDe at the late stage of biofilm formation. More importantly, once food is contaminated by L. monocytogenes EGDe, contamination proceeds independently of biofilm development and the initial level of contamination when food is stored at 4°C, even if contamination with L. monocytogenes EGDe was initially undetectable before storage.

Biofilm Formation and Associated Genes in Listeria Monocytogenes

2017 ◽  
Vol 12 (3) ◽  
Author(s):  
S. R. Warke ◽  
V. C. Ingle ◽  
N. V. Kurkure ◽  
P. A. Tembhurne ◽  
Minakshi Prasad ◽  
...  
Keyword(s):  
Public Health ◽  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Food Processing ◽  
Biofilm Formation ◽  
Milk Samples ◽  
Biofilm Production ◽  
Food Borne ◽  
Serious Infections ◽  
Microtiter Assay

Listeria monocytogenes, an opportunistic food borne pathogen can cause serious infections in immunocompromised individuals. L. monocytogenes is capable of producing biofilm on the surface of food processing lines and instruments.The biofilm transfers contamination to food products and impose risk to public health. In the present study biofilm producing ability of L. monocytogenes isolates were investigated phenotypically and genotypically by microtiter assay and multiplex PCR, respectively. Out of 38 L. monocytogenes isolates 14 were recovered from animal clinical cases, 12 bovine environment and 12 from milk samples. A total of 3 (21.42%) clinical, 2 (16.66%) environment and 3 (25%) milk samples respectively, revealed biofilm production in microtiter assay. Cumulative results showed that 23 (60.52%) out of 38 strains of L. monocytogenes were positive for luxS and flaA gene and 1 (2.63%) was positive only for the flaA gene.

Evaluation of Methods To Assess the Biofilm-Forming Ability of Listeria monocytogenes

2012 ◽  
Vol 75 (8) ◽  
pp. 1411-1417 ◽  
Author(s):  
ANTÓNIO LOURENÇO ◽  
FRANCISCO REGO ◽  
LUISA BRITO ◽  
JOSEPH F. FRANK
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
High Throughput ◽  
Biofilm Formation ◽  
High Throughput Screening ◽  
Screening Methods ◽  
Genetic Lineage ◽  
Production Lines ◽  
Genetic Characteristics

The contamination of ready-to-eat products with Listeria monocytogenes has been related to the presence of biofilms in production lines, as biofilms protect cells from chemical sanitizers. The ability of L. monocytogenes to produce biofilms is often evaluated using in vitro methodologies. This work aims to compare the most frequently used methodologies, including high-throughput screening methods based on microplates (crystal violet and the Calgary Biofilm Device) and methods based on CFU enumeration and microscopy after growth on stainless steel. Thirty isolates with diverse origins and genetic characteristics were evaluated. No (or low) correlations between methods were observed. The only significant correlation was found between the methods using stainless steel. No statistically significant correlation (P > 0.05) was detected among genetic lineage, serovar, and biofilm-forming ability. Because results indicate that biofilm formation is influenced by the surface material, the extrapolation of results from high-throughput methods using microplates to more industrially relevant surfaces should be undertaken with caution.

scholarly journals Persistent Listeria monocytogenes Isolates from a Poultry-Processing Facility Form More Biofilm but Do Not Have a Greater Resistance to Disinfectants than Sporadic Strains

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 250 ◽  
Author(s):  
Daniel Rodríguez-Campos ◽  
Cristina Rodríguez-Melcón ◽  
Carlos Alonso-Calleja ◽  
Rosa Capita
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Biofilm Formation ◽  
Benzalkonium Chloride ◽  
Bacterial Persistence ◽  
Crystal Violet Assay ◽  
Processing Facility ◽  
Poultry Processing ◽  
Sessile Cells ◽  
Selection Of

Some strains of Listeria monocytogenes can persist in food-processing environments, increasing the likelihood of the contamination of foodstuffs. To identify traits that contribute to bacterial persistence, a selection of persistent and sporadic L. monocytogenes isolates from a poultry-processing facility was investigated for biofilm-forming ability (crystal violet assay). The susceptibility of sessile cells to treatments (five minutes) with sodium hypochlorite having 10% active chlorine (SHY: 10,000 ppm, 25,000 ppm, and 50,000 ppm) and benzalkonium chloride (BZK: 2500 ppm, 10,000 ppm, and 25,000 ppm) was also studied. All isolates exhibited biofilm formation on polystyrene. Persistent strains showed larger (p < 0.001) biofilm formation (OD580 = 0.301 ± 0.097) than sporadic strains (OD580 = 0.188 ± 0.082). A greater susceptibility to disinfectants was observed for biofilms of persistent strains than for those of sporadic strains. The application of SHY reduced biofilms only for persistent strains. BZK increased OD580 in persistent strains (2500 ppm) and in sporadic strains (all concentrations). These results indicate that the use of BZK at the concentrations tested could represent a public health risk. Findings in this work suggest a link between persistence and biofilm formation, but do not support a relationship between persistence and the resistance of sessile cells to disinfectants.

Nanoengineered Superhydrophobic Surfaces to Prevent Adhesion of Listeria monocytogenes for Improved Food Safety

2020 ◽  
Vol 63 (5) ◽  
pp. 1401-1407
Author(s):  
Bog Eum Lee ◽  
Youngsang You ◽  
Won Choi ◽  
Eun-mi Hong ◽  
Marisa M. Wall ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Electrochemical Etching ◽  
Contact Angles ◽  
Bacterial Attachment ◽  
Superhydrophobic Surfaces ◽  
List Type ◽  
Ptfe Film ◽  
Steel Surfaces

HighlightsNanoporous superhydrophobic surfaces were fabricated using electrochemical etching and Teflon coating.Adhesion of Listeria monocytogenes to the nanoengineered stainless steel surfaces was reduced.Self-cleanable food-contact surfaces prevent bacterial attachment and subsequent biofilm formation.Abstract. Bacterial attachment on solid surfaces and subsequent biofilm formation is a significant problem in the food industry. Superhydrophobic surfaces have potential to prevent bacterial adhesion by minimizing the contact area between bacterial cells and the surface. In this study, stainless steel-based superhydrophobic surfaces were fabricated by manipulating nanostructures with electrochemical etching and polytetrafluoroethylene (PTFE) film. The formation of nanostructures on stainless steel surfaces was characterized by field emission scanning electron microscopy (FESEM). The stainless steel surfaces etched at 10 V for 5 min and at 10 V for 10 min with PTFE deposition resulted in average water contact angles of 154° ±4° with pore diameters of 50 nm. In addition, adhesion of Listeria monocytogenes was decreased by up to 99% compared to the bare substrate. These findings demonstrate the potential for the development of antibacterial surfaces by combining nanoporous patterns with PTFE films. Keywords: Electrochemical etching, PTFE, Nanoengineered surface, L. monocytogenes, Superhydrophobic.

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.

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