scholarly journals Disinfectant Susceptibility of Biofilm Formed by Listeria monocytogenes under Selected Environmental Conditions

2019 ◽  
Vol 7 (9) ◽  
pp. 280 ◽  
Author(s):  
Krzysztof Skowron ◽  
Ewa Wałecka-Zacharska ◽  
Katarzyna Grudlewska ◽  
Piotr Gajewski ◽  
Natalia Wiktorczyk ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Environmental Conditions ◽  
Biofilm Formation ◽  
Reference Strain ◽  
Bacterial Number ◽  
Food Borne ◽  
Nacl Concentration ◽  
Working Solution ◽  
Nutrients Availability ◽  
Strain Dependent

Listeria monocytogenes is a one of the most important food-borne pathogens. Its ability to form biofilm contributes to increased resistance to disinfectants and inefficient disinfection, posing a serious threat for the food industry, and in the end the consumer. The aim of this study was the comparison of the biofilm formation ability of L. monocytogenes strains on stainless steel, under different environmental conditions (temperature, pH, NaCl concentration, nutrients availability), and the assessment of biofilm susceptibility to disinfectants. The bactericidal activity of four disinfectants in two concentrations (100% and 50% of working solution) against biofilm was conducted on four clinical strains, four strains isolated from food and one reference strain ATCC 19111. It was found that biofilm susceptibility to disinfectants was influenced by environmental conditions. Biofilm susceptibility correlated with the decrease of temperature, pH, nutrients availability and salinity of the environment. The least sensitive to disinfectants was biofilm produced at pH = 4 (the bacterial number ranged from 0.25 log CFU × cm−2 to 1.72 log CFU × cm−2) whereas the most sensitive was biofilm produced at pH = 9 (5.16 log CFU × cm−2 to 7.84 log CFU × cm−2). Quatosept was the most effective disinfectant, regardless of the conditions. In conclusion, biofilm susceptibility to disinfectants is strain-dependent and is affected by environmental conditions.

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 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 Loss of Flagellum-Based Motility by Listeria monocytogenes Results in Formation of Hyperbiofilms

2008 ◽  
Vol 190 (17) ◽  
pp. 6030-6034 ◽  
Author(s):  
Tatsaporn Todhanakasem ◽  
Glenn M. Young
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
High Density ◽  
Gram Positive ◽  
Surface Attachment ◽  
Bacterial Surface ◽  
Food Borne ◽  
Biofilm Development

ABSTRACT Biofilm formation by the gram-positive, motile, food-borne pathogen Listeria monocytogenes was demonstrated to occur by an ordered series of stages. Biofilm development involves flagellum-based motility, which when blocked decreases initial bacterial surface attachment but subsequently leads to the formation of hyperbiofilms, surface-attached communities reaching high density.

scholarly journals A Putative ABC Transporter Is Involved in Negative Regulation of Biofilm Formation by Listeria monocytogenes

2008 ◽  
Vol 74 (24) ◽  
pp. 7675-7683 ◽  
Author(s):  
Xinna Zhu ◽  
Fei Long ◽  
Yonghui Chen ◽  
Susanne Knøchel ◽  
Qunxin She ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Abc Transporter ◽  
Biofilm Formation ◽  
Negative Regulation ◽  
Atp Binding ◽  
Coding Region ◽  
Gene Encoding ◽  
Food Borne ◽  
Atp Binding Protein

ABSTRACT Listeria monocytogenes may persist for long periods in food processing environments. In some instances, this may be due to aggregation or biofilm formation. To investigate the mechanism controlling biofilm formation in the food-borne pathogen L. monocytogenes, we characterized LM-49, a mutant with enhanced ability of biofilm formation generated via transposon Tn917 mutagenesis of L. monocytogenes 4b G. In this mutant, a Tn917 insertion has disrupted the coding region of the gene encoding a putative ATP-binding cassette (ABC) transporter permease identical to Lmof2365_1771 (a putative ABC transporter permease) presented in the sequenced strain L. monocytogenes strain 4b F2365. This disrupted gene, denoted lm.G_1771, encoded a protein with 10 transmembrane helixes. The revertant, LM-49RE, was obtained by replacing lm.G_1771::Tn917 with lm.G_1771 via homologous recombination. We found that LM-49RE formed the same amount of biofilm biomass as the wild-type strain. Furthermore, transcription of the downstream lm.G_1770 gene was not influenced by the upstream Tn917 insertion, and the presence of Tn917 has no effect on biofilm formation. These results suggest that lm.G_1771 was solely responsible for the negative regulation of biofilm formation by L. monocytogenes 4b G. The immediate gene upstream of lm.G_1771 encoded an ATP-binding protein. Bioinformatics analysis suggested that these two genes were organized into an operon and that their proteins formed an export ABC transporter. Here, we report the characterization of the mutant and identification of a novel ABC transporter that functions in negative regulation of biofilm formation in L. monocytogenes.

scholarly journals Characteristics of Listeria monocytogenes Strains Isolated from Milk and Humans and the Possibility of Milk-Borne Strains Transmission

2019 ◽  
Vol 68 (3) ◽  
pp. 353-369
Author(s):  
KRZYSZTOF SKOWRON ◽  
EWA WAŁECKA-ZACHARKSA ◽  
KATARZYNA GRUDLEWSKA ◽  
NATALIA WIKTORCZYK ◽  
AGNIESZKA KACZMAREK ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Human Blood ◽  
Biofilm Formation ◽  
Dairy Products ◽  
Virulence Genes ◽  
Reference Strain ◽  
Drug Susceptibility ◽  
Strain Atcc ◽  
Erythromycin Resistance ◽  
Dairy Processing

Listeria monocytogenes is the etiological factor of listeriosis. The main source of these organisms is food, including dairy products. The aim was to determine the multiple correlations between the drug susceptibility, virulence genes (VGs), and biofilm formation on silicone teat cups of milk-borne and human L. monocytogenes strains. The spread of L. monocytogenes via contaminated teat rubbers was assessed. The L. monocytogenes strains recovered from milk (18), human blood (10), and the reference strain ATCC®19111™ were used in the study. Penicillin resistance was the most prevalent resistance in the milk isolates (n = 8; 44.4%), whereas among clinical strains erythromycin resistance was predominating – (n = 6; 60%). The most frequent VGs among strains isolated from milk were hlyA (100%) and plcB (100%) whereas in strains isolated from blood – hlyA (100%) and prfA (90%). All tested VGs were present in 50% of blood isolates and 11% of milk-borne strains. The strains isolated from milk formed a significantly stronger biofilm. The strains with more numerous virulence genes were resistant to more antibiotics and formed a stronger biofilm. It was shown that contaminated teat cups might contribute to the transmission of L. monocytogenes in the herd. It seems reasonable to monitor the occurrence of L. monocytogenes biofilm in a dairy processing environment.

scholarly journals Role of PBPD1 in Stimulation of Listeria monocytogenes Biofilm Formation by Subminimal Inhibitory β-Lactam Concentrations

2014 ◽  
Vol 58 (11) ◽  
pp. 6508-6517 ◽  
Author(s):  
Uyen T. Nguyen ◽  
Hanjeong Harvey ◽  
Andrew J. Hogan ◽  
Alexandria C. F. Afonso ◽  
Gerard D. Wright ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Regulatory System ◽  
Content Type ◽  
Food Borne ◽  
Approved Drugs ◽  
Biofilm Development ◽  
Fda Approved Drugs ◽  
Stimulation Of

ABSTRACTDisinfectant-tolerantListeria monocytogenesbiofilms can colonize surfaces that come into contact with food, leading to contamination and, potentially, food-borne illnesses. To better understand the process ofL. monocytogenesbiofilm formation and dispersal, we screened 1,120 off-patent FDA-approved drugs and identified several that modulateListeriabiofilm development. Among the hits were more than 30 β-lactam antibiotics, with effects ranging from inhibiting (≤50%) to stimulating (≥200%) biofilm formation compared to control. Most β-lactams also dispersed a substantial proportion of established biofilms. This phenotype did not necessarily involve killing, as >50% dispersal could be achieved with concentrations as low as 1/20 of the MIC of some cephalosporins. Penicillin-binding protein (PBP) profiling using a fluorescent penicillin analogue showed similar inhibition patterns for most β-lactams, except that biofilm-stimulatory drugs did not bind PBPD1, a low-molecular-weightd,d-carboxypeptidase. Compared to the wild type, apbpD1mutant had an attenuated biofilm response to stimulatory β-lactams. The cephalosporin-responsive CesRK two-component regulatory system, whose regulon includes PBPs, was not required for the response. The requirement for PBPD1 activity for β-lactam stimulation ofL. monocytogenesbiofilms shows that the specific set of PBPs that are inactivated by a particular drug dictates whether a protective biofilm response is provoked.

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 Importance of SigB for Listeria monocytogenes Static and Continuous-Flow Biofilm Formation and Disinfectant Resistance

2010 ◽  
Vol 76 (23) ◽  
pp. 7854-7860 ◽  
Author(s):  
Stijn van der Veen ◽  
Tjakko Abee
Keyword(s):  
Listeria Monocytogenes ◽  
Peracetic Acid ◽  
Continuous Flow ◽  
Biofilm Formation ◽  
Benzalkonium Chloride ◽  
Antimicrobial Agents ◽  
Resistance Mechanisms ◽  
Stress Response Genes ◽  
Food Borne

ABSTRACT Listeria monocytogenes is a food-borne pathogen that is able to form biofilms in food processing facilities. Biofilms are generally more resistant to antimicrobial agents, making it difficult to eradicate them during cleanup procedures. So far, little is known about the function of stress resistance mechanisms in biofilm formation and their resistance to disinfectants. In this study, we investigated the role of sigB, which encodes a major transcriptional regulator of stress response genes, in L. monocytogenes static and continuous-flow biofilm formation and its function in the resistance of biofilm cells to the disinfectants benzalkonium chloride and peracetic acid. Quantitative real-time PCR and promoter reporter studies showed that sigB is activated in static and continuous-flow biofilms. Biofilm formation studies using an in-frame sigB deletion mutant and complementation mutant showed that the presence of SigB is required to obtain wild-type levels of both static and continuous-flow biofilms. Finally, disinfection treatments of planktonically grown cells and cells dispersed from static and continuous-flow biofilms showed that SigB is involved in the resistance of both planktonic cells and biofilms to the disinfectants benzalkonium chloride and peracetic acid.

Inhibition of Bacterial Biofilm Formation by Phytotherapeutics with Focus on Overcoming Antimicrobial Resistance

2020 ◽  
Vol 26 (24) ◽  
pp. 2807-2816 ◽  
Author(s):  
Yun Su Jang ◽  
Tímea Mosolygó
Keyword(s):  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Bacterial Biofilm ◽  
Experimental Investigations ◽  
Resistant Bacteria ◽  
Salmonella Spp ◽  
Food Borne ◽  
High Prevalence ◽  
Scientific Attention

: Bacteria within biofilms are more resistant to antibiotics and chemical agents than planktonic bacteria in suspension. Treatment of biofilm-associated infections inevitably involves high dosages and prolonged courses of antimicrobial agents; therefore, there is a potential risk of the development of antimicrobial resistance (AMR). Due to the high prevalence of AMR and its association with biofilm formation, investigation of more effective anti-biofilm agents is required. : From ancient times, herbs and spices have been used to preserve foods, and their antimicrobial, anti-biofilm and anti-quorum sensing properties are well known. Moreover, phytochemicals exert their anti-biofilm properties at sub-inhibitory concentrations without providing the opportunity for the emergence of resistant bacteria or harming the host microbiota. : With increasing scientific attention to natural phytotherapeutic agents, numerous experimental investigations have been conducted in recent years. The present paper aims to review the articles published in the last decade in order to summarize a) our current understanding of AMR in correlation with biofilm formation and b) the evidence of phytotherapeutic agents against bacterial biofilms and their mechanisms of action. The main focus has been put on herbal anti-biofilm compounds tested to date in association with Staphylococcus aureus, Pseudomonas aeruginosa and food-borne pathogens (Salmonella spp., Campylobacter spp., Listeria monocytogenes and Escherichia coli).

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