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.

scholarly journals Variation in Biofilm Formation among Strains of Listeria monocytogenes

2003 ◽  
Vol 69 (12) ◽  
pp. 7336-7342 ◽  
Author(s):  
Monica K. Borucki ◽  
Jason D. Peppin ◽  
David White ◽  
Frank Loge ◽  
Douglas R. Call
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Biofilm Formation ◽  
Disease Incidence ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Cell Adherence ◽  
Milk Samples ◽  
Bulk Milk ◽  
Food Borne

ABSTRACT Contamination of food by Listeria monocytogenes is thought to occur most frequently in food-processing environments where cells persist due to their ability to attach to stainless steel and other surfaces. Once attached these cells may produce multicellular biofilms that are resistant to disinfection and from which cells can become detached and contaminate food products. Because there is a correlation between virulence and serotype (and thus phylogenetic division) of L. monocytogenes, it is important to determine if there is a link between biofilm formation and disease incidence for L. monocytogenes. Eighty L. monocytogenes isolates were screened for biofilm formation to determine if there is a robust relationship between biofilm formation, phylogenic division, and persistence in the environment. Statistically significant differences were detected between phylogenetic divisions. Increased biofilm formation was observed in Division II strains (serotypes 1/2a and 1/2c), which are not normally associated with food-borne outbreaks. Differences in biofilm formation were also detected between persistent and nonpersistent strains isolated from bulk milk samples, with persistent strains showing increased biofilm formation relative to nonpersistent strains. There were no significant differences detected among serotypes. Exopolysaccharide production correlated with cell adherence for high-biofilm-producing strains. Scanning electron microscopy showed that a high-biofilm-forming strain produced a dense, three-dimensional structure, whereas a low-biofilm-forming strain produced a thin, patchy biofilm. These data are consistent with data on persistent strains forming biofilms but do not support a consistent relationship between enhanced biofilm formation and disease incidence.

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.

scholarly journals Biofilm Formation among Clinical and Food Isolates ofListeria monocytogenes

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Joana Barbosa ◽  
Sandra Borges ◽  
Ruth Camilo ◽  
Rui Magalhães ◽  
Vânia Ferreira ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Listeria Monocytogenes ◽  
Osmotic Stress ◽  
Environmental Conditions ◽  
Biofilm Formation ◽  
Stress Conditions ◽  
Acidic Stress ◽  
Biofilm Production ◽  
Oflisteria Monocytogenes ◽  
Clinical Cases

Objective. A total of 725Listeria monocytogenesisolates, 607 from various foods and 118 from clinical cases of listeriosis, were investigated concerning their ability to form biofilms, at 4°C during 5 days and at 37°C during 24 h.Methods. Biofilm production was carried out on polystyrene tissue culture plates. FiveL. monocytogenesisolates were tested for biofilm formation after being exposed to acidic and osmotic stress conditions.Results. Significant differences (P<0.01) between clinical and food isolates were observed. At 37°C for 24 h, most food isolates were classified as weak or moderate biofilm formers whereas all the clinical isolates were biofilm producers, although the majority were weak. At 4°C during 5 days, 65 and 59% isolates, from food and clinical cases, respectively, were classified as weak. After both sublethal stresses, at 37°C just one of the five isolates tested was shown to be more sensitive to subsequent acidic exposure. However, at 4°C both stresses did not confer either sensitivity or resistance.Conclusions. Significant differences between isolates origin, temperature, and sublethal acidic stress were observed concerning the ability to form biofilms. Strain, origin, and environmental conditions can determine the level of biofilm production byL. monocytogenesisolates.

scholarly journals Role of Extracellular DNA during Biofilm Formation by Listeria monocytogenes

2010 ◽  
Vol 76 (7) ◽  
pp. 2271-2279 ◽  
Author(s):  
Morten Harmsen ◽  
Martin Lappann ◽  
Susanne Kn�chel ◽  
S�ren Molin
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Extracellular Dna ◽  
Central Component ◽  
High Molecular Weight Dna ◽  
Food Borne ◽  
Attachment Sites ◽  
Sperm Dna ◽  
Salmon Sperm Dna

ABSTRACT Listeria monocytogenes is a food-borne pathogen that is capable of living in harsh environments. It is believed to do this by forming biofilms, which are surface-associated multicellular structures encased in a self-produced matrix. In this paper we show that in L. monocytogenes extracellular DNA (eDNA) may be the only central component of the biofilm matrix and that it is necessary for both initial attachment and early biofilm formation for 41 L. monocytogenes strains that were tested. DNase I treatment resulted in dispersal of biofilms, not only in microtiter tray assays but also in flow cell biofilm assays. However, it was also demonstrated that in a culture without eDNA, neither Listeria genomic DNA nor salmon sperm DNA by itself could restore the capacity to adhere. A search for additional necessary components revealed that peptidoglycan (PG), specifically N-acetylglucosamine (NAG), interacted with the DNA in a manner which restored adhesion. If a short DNA fragment (less than approximately 500 bp long) was added to an eDNA-free culture prior to addition of genomic or salmon sperm DNA, adhesion was prevented, indicating that high-molecular-weight DNA is required for adhesion and that the number of attachment sites on the cell surface can be saturated.

scholarly journals Microtiter Plate Assay for Assessment of Listeria monocytogenes Biofilm Formation

2002 ◽  
Vol 68 (6) ◽  
pp. 2950-2958 ◽  
Author(s):  
D. Djordjevic ◽  
M. Wiedmann ◽  
L. A. McLandsborough
Keyword(s):  
Listeria Monocytogenes ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Cellular Growth ◽  
Epifluorescence Microscopy ◽  
Simple Method ◽  
Microtiter Plate Assay ◽  
Plate Assay ◽  
Biofilm Production

ABSTRACT Listeria monocytogenes has the ability to form biofilms on food-processing surfaces, potentially leading to food product contamination. The objective of this research was to standardize a polyvinyl chloride (PVC) microtiter plate assay to compare the ability of L. monocytogenes strains to form biofilms. A total of 31 coded L. monocytogenes strains were grown in defined medium (modified Welshimer's broth) at 32°C for 20 and 40 h in PVC microtiter plate wells. Biofilm formation was indirectly assessed by staining with 1% crystal violet and measuring crystal violet absorbance, using destaining solution. Cellular growth rates and final cell densities did not correlate with biofilm formation, indicating that differences in biofilm formation under the same environmental conditions were not due to growth rate differences. The mean biofilm production of lineage I strains was significantly greater than that observed for lineage II and lineage III strains. The results from the standardized microtiter plate biofilm assay were also compared to biofilm formation on PVC and stainless steel as assayed by quantitative epifluorescence microscopy. Results showed similar trends for the microscopic and microtiter plate assays, indicating that the PVC microtiter plate assay can be used as a rapid, simple method to screen for differences in biofilm production between strains or growth conditions prior to performing labor-intensive microscopic analyses.

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.

Listeria monocytogenes Persistence in Food-Associated Environments: Epidemiology, Strain Characteristics, and Implications for Public Health

2014 ◽  
Vol 77 (1) ◽  
pp. 150-170 ◽  
Author(s):  
V. FERREIRA ◽  
M. WIEDMANN ◽  
P. TEIXEIRA ◽  
M. J. STASIEWICZ
Keyword(s):  
Public Health ◽  
Listeria Monocytogenes ◽  
Food Processing ◽  
Current Knowledge ◽  
Molecular Subtype ◽  
Special Focus ◽  
Quantitative Microbial Risk Assessment ◽  
Genetic Characteristics ◽  
Economic Implications ◽  
Processing Plants

Over the last 10 to 15 years, increasing evidence suggests that persistence of Listeria monocytogenes in food processing plants for years or even decades is an important factor in the transmission of this foodborne pathogen and the root cause of a number of human listeriosis outbreaks. L. monocytogenes persistence in other food-associated environments (e.g., farms and retail establishments) may also contribute to food contamination and transmission of the pathogen to humans. Although L. monocytogenes persistence is typically identified through isolation of a specific molecular subtype from samples collected in a given environment over time, formal (statistical) criteria for identification of persistence are undefined. Environmental factors (e.g., facilities and equipment that are difficult to clean) have been identified as key contributors to persistence; however, the mechanisms are less well understood. Although some researchers have reported that persistent strains possess specific characteristics that may facilitate persistence (e.g., biofilm formation and better adaptation to stress conditions), other researchers have not found significant differences between persistent and nonpersistent strains in the phenotypic characteristics that might facilitate persistence. This review includes a discussion of our current knowledge concerning some key issues associated with the persistence of L. monocytogenes, with special focus on (i) persistence in food processing plants and other food-associated environments, (ii) persistence in the general environment, (iii) phenotypic and genetic characteristics of persistent strains, (iv) niches, and (v) public health and economic implications of persistence. Although the available data clearly indicate that L. monocytogenes persistence at various stages of the food chain contributes to contamination of finished products, continued efforts to quantitatively integrate data on L. monocytogenes persistence (e.g., meta-analysis or quantitative microbial risk assessment) will be needed to advance our understanding of persistence of this pathogen and its economic and public health impacts.

scholarly journals Distribution of serotypes of Listeria monocytogenes in chicken meats in Turkey

2020 ◽  
Vol 70 (4) ◽  
pp. 1859
Author(s):  
S. SAHIN ◽  
R. KALIN ◽  
MN MOGULKOC
Keyword(s):  
Public Health ◽  
Listeria Monocytogenes ◽  
Large Scale ◽  
Meat Products ◽  
Health Problems ◽  
Chicken Meat ◽  
Serotype Distribution ◽  
Food Borne ◽  
Serotype 1 ◽  
Food Borne Infections

Listeria monocytogenes is one of the important causes of food-borne infections. This study was conducted to determine the presence of L. monocytogenes and its serotype distribution in a total of 400 packaged chicken meat products (drumstick, breast, wing, and whole chicken) from different national companies. L. monocytogenes contamination was detected in 26.5% (106 in 400) of all samples when the products considered, drumsticks, breasts, wings, and whole chickens showed 47%, 15%, 35, and 9% positivity respectively. Four important serotypes of L. monocytogenes in human listeriosis (1/2a, 1/2b, 1/2c and 4b) were identified, and serotype 1/2a (94.3%) was determined as predominant in packaged chicken meats. The present study revealed that L. monocytogenes 1/2a serotype is prevalent in chicken meats and this may cause public health problems in Turkey. Further studies in poultry meats should be conducted on a large scale such as regional or national big markets to determine the presence of the pathogen and its dominant serotypes.

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