Biofilm Formation Ability of Listeria monocytogenes Isolates from Raw Ready-to-Eat Seafood

2009 ◽  
Vol 72 (7) ◽  
pp. 1476-1480 ◽  
Author(s):  
HAJIME TAKAHASHI ◽  
SATOKO MIYA ◽  
KAZUNORI IGARASHI ◽  
TAKAYUKI SUDA ◽  
SHINTARO KURAMOTO ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Environmental Factors ◽  
Biofilm Formation ◽  
Foodborne Pathogen ◽  
Microtiter Plate ◽  
Processing Plant ◽  
High Incidence ◽  
Sporadic Cases ◽  
Different Levels

Listeria monocytogenes is of great concern as a foodborne pathogen. Many ready-to-eat foods are widely contaminated with this organism and have caused listeriosis outbreaks and sporadic cases in many countries. In Japan, there is a high incidence of L. monocytogenes contamination, specifically in raw ready-to-eat seafood. Identical L. monocytogenes subtypes have been isolated repeatedly from samples of food manufactured at a given store or processing plant, and researchers suspected that certain L. monocytogenes isolates have formed biofilms at these sites. A microtiter plate biofilm formation assay was conducted, and all raw ready-to-eat seafood isolates tested were able to form biofilms to various degrees. Biofilm formation by L. monocytogenes isolates of lineage I was significantly greater (P = 0.000) than that by isolates of lineage II. However, isolates of clonal lineages formed different levels of biofilms, indicating that the ability to form a biofilm is affected positively or negatively by environmental factors.

Biofilm Formation under Different Temperature Conditions by a Single Genotype of Persistent Listeria monocytogenes Strains

2014 ◽  
Vol 77 (1) ◽  
pp. 133-140 ◽  
Author(s):  
YOSHITSUGU OCHIAI ◽  
FUMIYA YAMADA ◽  
MARIKO MOCHIZUKI ◽  
TAKASHI TAKANO ◽  
RYO HONDO ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Environmental Factors ◽  
Biofilm Formation ◽  
Processing Plant ◽  
Dependent Manner ◽  
Temperature Dependent ◽  
Significant Difference ◽  
Single Genotype ◽  
Serotype 1 ◽  
Retail Shop

Some Listeria monocytogenes strains, termed persistent strains, originate from the same processing plant and have the ability to survive and grow over extended periods of time at contamination sources. In order to evaluate biofilm formation by such persistent strains, we isolated the pathogen from chicken samples collected from the same retail shop in repeated visits over 6 months. Strains that were of serotype 1/2b and were assigned to the same genotype by multi-virulence-locus sequence typing analysis were isolated on repeated occasions from December 1997 to June 1998 and thus were defined as persistent strains. In the present study, biofilm formation by the persistent strains was evaluated using microplates at 30 and 37°C. The biofilm-forming capability was measured after cells attaching to the microplate well were stained with crystal violet. Comparison of biofilm formation at 30°C among the persistent strains showed that a significantly higher amount of the stain was obtained from the persistent strains isolated from December to March than from those isolated from April to June. However, no significant difference in biofilm formation at 30°C was observed between persistent and nonpersistent groups of L. monocytogenes strains. In contrast, biofilm formation at 37°C was consistent among the persistent strains, and they produced significantly more biofilm at 37°C than did the nonpersistent strains. The persistent strains were also found to change their biofilm-forming ability in a temperature-dependent manner, which may suggest that the persistent strains alter their biofilm formation in response to changing environmental factors.

scholarly journals The biofilm formation ability of Listeria monocytogenes isolated from meat, poultry, fish and processing plant environments is related to serotype and pathogenic profile of the strains

2012 ◽  
Vol 2 (1) ◽  
pp. 12 ◽  
Author(s):  
Domenico Meloni ◽  
Roberta Mazza ◽  
Francesca Piras ◽  
Sonia Lamon ◽  
Simonetta Gianna Consolati ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Target Genes ◽  
Microtiter Plate ◽  
Food Sources ◽  
Processing Plant ◽  
Pathogenic Potential ◽  
Microtiter Plate Assay ◽  
Processing Plants

In the present study, the relationships between serotype, pathogenic profile and <em>in vitro</em> biofilm formation of 106 <em>Listeria monocytogenes</em> strains, having no epidemiological correlation and isolated from different environmental and food sources, were analyzed. The quantitative assessment of the <em>in vitro</em> biofilm formation was carried out by using a microtiter plate assay with spectrophotometric reading (OD620). The isolates were also submitted to serogrouping using the target genes <em>lmo0737</em>, <em>lmo1118, ORF2819, ORF2110, prs</em>, and to the evaluation of the presence of the following virulence genes: <em>prfA, hlyA, rrn, inlA, inlB, iap, plcA, plcB, actA</em> and <em>mpl</em>, by multiplex PCRs. The 62% of the strains showed weak or moderate <em>in vitro</em> ability in biofilm formation, in particular serotypes 1/2b and 4b, frequently associated with sporadic or epidemic listeriosis cases. The 25% of these isolates showed polymorphism for the <em>actA</em> gene, producing a fragment of 268-bp instead of the expected 385-bp. The deletion of nucleotides in this gene seems to be related to enhanced virulence properties among these strains. Strains belonging to serotypes associated with human infections and characterized by pathogenic potential are capable to persist within the processing plants forming biofilm.

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.

scholarly journals Identification of Listeria monocytogenes Contamination in a Ready-to-Eat Meat Processing Plant in China

2021 ◽  
Vol 12 ◽  
Author(s):  
Hongzhi Zhang ◽  
Fengxia Que ◽  
Biyao Xu ◽  
Linjun Sun ◽  
Yanqi Zhu ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Raw Materials ◽  
Case Fatality ◽  
Etiologic Agent ◽  
Health Concern ◽  
Plate Count ◽  
Processing Plant ◽  
Meat Processing ◽  
Processing Plants

Listeria monocytogenes is the etiologic agent of listeriosis, which remains a significant public health concern in many countries due to its high case-fatality rate. The constant risk of L. monocytogenes transmission to consumers remains a central challenge in the food production industry. At present, there is very little known about L. monocytogenes contamination in ready-to-eat (RTE) processing plants in China. In this study, L. monocytogenes in an RTE meat processing plant in Shanghai municipality was characterized using pulsed-field gel electrophoresis (PFGE) and whole genome sequencing (WGS). Furthermore, the biofilm formation ability of the pathogen was also tested. Results revealed that L. monocytogenes isolates were present in 12 samples out of the 48 samples investigated. Most of them (66.7%, 8/12) were identified from the processing facilities irrespective of observed hygiene levels of aerobic plate count (APC) and coliforms. Coliforms were present in only one processing area. ST5 (1/2b) isolates were predominant (83.3%, 10/12) and were identified in two dominant pulsotypes (PTs) (three in PT3 and seven in PT4, respectively). Results of the core-genome multi-locus sequence typing (cgMLST) showed that ST5 in three PTs (PT1, PT3, and PT4) had 0–8 alleles, which confirmed that clonal transmission occurred in the RTE meat processing facilities. In addition, the biofilm formation test confirmed that the isolates from the processing facilities could form biofilms, which helped them colonize and facilitate persistence in the environment. These results indicated that common sanitation procedures regularly applied in the processing environment were efficient but not sufficient to remove L. monocytogenes isolates, especially biofilm of L. monocytogenes. Furthermore, the ST5 isolates in this study exhibited 12 alleles with one ST5 clinical isolate, which contributes to the understanding of the potential pathogenic risk that L. monocytogenes in RTE meat processing equipment posed to consumers. Therefore, strong hygienic measures, especially sanitation procedures for biofilms eradication, should be implemented to ensure the safety of raw materials. Meanwhile, continuous surveillance might be vital for the prevention and control of listeriosis caused by L. monocytogenes.

scholarly journals Novel Cadmium Resistance Determinant in Listeria monocytogenes

2016 ◽  
Vol 83 (5) ◽  
Author(s):  
Cameron Parsons ◽  
Sangmi Lee ◽  
Victor Jayeola ◽  
Sophia Kathariou
Keyword(s):  
Heavy Metals ◽  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Galleria Mellonella ◽  
Foodborne Pathogen ◽  
Whole Genome ◽  
Cadmium Resistance ◽  
Content Type ◽  
Content Model ◽  
Tolerance To Cadmium

ABSTRACT Listeria monocytogenes is a foodborne pathogen that can cause severe disease (listeriosis) in susceptible individuals. It is ubiquitous in the environment and often exhibits resistance to heavy metals. One of the determinants that enables Listeria to tolerate exposure to cadmium is the cadAC efflux system, with CadA being a P-type ATPase. Three different cadA genes (designated cadA1 to cadA3) were previously characterized in L. monocytogenes. A novel putative cadmium resistance gene (cadA4) was recently identified through whole-genome sequencing, but experimental confirmation for its involvement in cadmium resistance is lacking. In this study, we characterized cadA4 in L. monocytogenes strain F8027, a cadmium-resistant strain of serotype 4b. By screening a mariner-based transposon library of this strain, we identified a mutant with reduced tolerance to cadmium and that harbored a single transposon insertion in cadA4. The tolerance to cadmium was restored by genetic complementation with the cadmium resistance cassette (cadA4C), and enhanced cadmium tolerance was conferred to two unrelated cadmium-sensitive strains via heterologous complementation with cadA4C. Cadmium exposure induced cadA4 expression, even at noninhibitory levels. Virulence assessments in the Galleria mellonella model suggested that a functional cadA4 suppressed virulence, potentially promoting commensal colonization of the insect larvae. Biofilm assays suggested that cadA4 inactivation reduced biofilm formation. These data not only confirm cadA4 as a novel cadmium resistance determinant in L. monocytogenes but also provide evidence for roles in virulence and biofilm formation. IMPORTANCE Listeria monocytogenes is an intracellular foodborne pathogen causing the disease listeriosis, which is responsible for numerous hospitalizations and deaths every year. Among the adaptations that enable the survival of Listeria in the environment are the abilities to persist in biofilms, grow in the cold, and tolerate toxic compounds, such as heavy metals. Here, we characterized a novel determinant that was recently identified on a larger mobile genetic island through whole-genome sequencing. This gene (cadA4) was found to be responsible for cadmium detoxification and to be a divergent member of the Cad family of cadmium efflux pumps. Virulence assessments in a Galleria mellonella model suggested that cadA4 may suppress virulence. Additionally, cadA4 may be involved in the ability of Listeria to form biofilms. Beyond the role in cadmium detoxification, the involvement of cadA4 in other cellular functions potentially explains its retention and wide distribution in L. monocytogenes.

Biofilm Formation and Resistance to Benzalkonium Chloride in Listeria monocytogenes Isolated from a Fish Processing Plant

2013 ◽  
Vol 76 (7) ◽  
pp. 1179-1186 ◽  
Author(s):  
HIROMI NAKAMURA ◽  
KOH-ICHI TAKAKURA ◽  
YOSHIAKI SONE ◽  
YASUYUKI ITANO ◽  
YOSHIKAZU NISHIKAWA
Keyword(s):  
Listeria Monocytogenes ◽  
Benzalkonium Chloride ◽  
Extracellular Polymeric Substances ◽  
Foodborne Pathogen ◽  
Maximum Activity ◽  
Processing Plant ◽  
Fish Processing ◽  
Glucose Content ◽  
Transient Strain ◽  
Life Threatening Illness

Listeria monocytogenes is a foodborne pathogen that causes the potentially life-threatening illness listeriosis. Previously, a few clones of L. monocytogenes persisting in a cold-smoked fish processing plant were isolated from the plant's products continuously. To evaluate the role of biofilms in the persistence of L. monocytogenes strains specific to this plant, the abilities of the persistent strain (PS) and transient strain (TS) of L. monocytogenes found in this plant to form biofilms were compared, as was resistance to the sanitizing effects of benzalkonium chloride (BC). The PS produced more biofilm than the TS in 48 h. The half-maximal effective concentration (EC50), the BC concentration at which the ATP bioluminescence of each bacterial strain decreased by 50% relative to its maximum activity, was about 150-fold higher in the PS than in the TS. In contrast, when these values were measured in organisms in a planktonic state, the EC50 of the PS was only 2.2-fold higher than that of the TS. Extracellular polymeric substances (EPS) were extracted from biofilms, and the glucose content of these biofilms was determined with the phenol–sulfuric acid method to estimate the quantity of EPS. The total amount of EPS in the PS biofilm was higher than that in the TS biofilm. These findings suggest that the PS produces greater amounts of biofilm and EPS than the TS, which results in greater resistance of the PS to disinfectants. The persistence of the strain in the fish processing plant might be attributable to these properties.

Removal of Listeria monocytogenes Biofilms with Bacteriophage P100†

2010 ◽  
Vol 73 (8) ◽  
pp. 1519-1524 ◽  
Author(s):  
KAMLESH A. SONI ◽  
RAMAKRISHNA NANNAPANENI
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Biofilm Formation ◽  
Foodborne Pathogen ◽  
Growth Conditions ◽  
Cell Populations ◽  
Wide Range ◽  
Serotype 1 ◽  
Stainless Steel Coupon ◽  
Steel Coupon

Listeria monocytogenes is an important foodborne pathogen with a persistent ability to form biofilm matrices in the food processing environments. In this study, we have determined the ability of bacteriophage P100 to reduce L. monocytogenes cell populations under biofilm conditions by using 21 L. monocytogenes strains representing 13 different serotypes. There were considerable differences in the ability of various strains of L. monocytogenes to form biofilms, with strains of serotype 1/2a showing maximum biofilm formation. Irrespective of the serotype, growth conditions, or biofilm levels, the phage P100 treatment significantly reduced L. monocytogenes cell populations under biofilm conditions. On the stainless steel coupon surface, there was a 3.5- to 5.4-log/cm2 reduction in L. monocytogenes cells by phage treatment. These findings illustrate that phage P100 is active against a wide range of L. monocytogenes strains in biofilm conditions.

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