Growth and Survival of Attached Listeria on Lettuce and Stainless Steel Varies by Strain and Surface Type

2021 ◽  
Author(s):  
Lisa Gorski ◽  
Samarpita Walker ◽  
Kelly F Romanolo ◽  
Sophia Kathariou
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Fluorescent Protein ◽  
Foodborne Pathogen ◽  
Surface Growth ◽  
Growth And Survival ◽  
Abiotic Surfaces ◽  
Serotype 1 ◽  
Serotype 4B ◽  
Green Fluorescent

The foodborne pathogen Listeria monocytogenes lives as a saprophyte in nature and can adhere to and grows on surfaces as diverse as leaves, sediment, and stainless steel. To discern the mechanisms used by L. monocytogenes for attachment and growth on various surfaces, we studied interactions between the pathogen on lettuce and stainless steel. A panel of 24 strains (23 of Listeria monocytogenes and 1 L. innocua ) was screened for attachment and growth on lettuce at 4 o C and 25 o C and on stainless steel at 10 o C and 37 o C. Overnight growth of attached cells resulted in a 0 – 3 log increase on lettuce, depending on the strain and the temperature. Among the worst performing strains on lettuce were two from a large cantaloupe outbreak, indicating that factors important for interactions with cantaloupe may be different from those required on lettuce tissue. Strains that grew the best on lettuce belonged to serotypes 1/2a, 1/2b, and 4b and were from cheese, potatoes, and water/sediment near produce fields. Confocal microscopy of L. monocytogenes tagged with constitutively expressed green fluorescent protein indicated associations with the cut edges and veins of lettuce leaves. On stainless steel coupons, there was a 5 – 7 log increase at 10 o C after 7 d and a 4 – 7 log increase at 37 o C after 40 h. Statistically, surface growth on stainless steel was better for serotype 1/2a than for serotype 4b strains, even though certain serotype 4b strains grew well on the coupons. The latter included strains that originated from produce and water/sediment. Some strains were fit in both environments, while others showed variability between the two different surfaces. Further analysis of these strains should reveal molecular factors needed for adherence and surface growth of L. monocytogenes on different biotic and abiotic surfaces.

scholarly journals Genome Sequence of Listeria monocytogenes Strain F4244, a 4b Serotype

2017 ◽  
Vol 5 (49) ◽  
Author(s):  
Taylor W. Bailey ◽  
Naila C. do Nascimento ◽  
Arun K. Bhunia
Keyword(s):  
Listeria Monocytogenes ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Genome Sequence ◽  
Illumina Sequencing ◽  
Foodborne Pathogen ◽  
Whole Genome ◽  
Content Type ◽  
Serotype 1 ◽  
Serotype 4B

ABSTRACT Listeria monocytogenes is an opportunistic invasive foodborne pathogen. Here, we performed whole-genome sequencing of L. monocytogenes strain F4244 (serotype 4b) using Illumina sequencing. The sequence showed 94.5% identity with strain F2365, serotype 4b, and 90.6% with EGD-e, serotype 1/2a.

scholarly journals Draft Genome Sequences of Listeria monocytogenes Serotype 4b Strains 944 and 2993 and Serotype 1/2c Strains 198 and 2932

2016 ◽  
Vol 4 (3) ◽  
Author(s):  
Aidan Casey ◽  
Olivia McAuliffe ◽  
Edward M. Fox ◽  
Dara Leong ◽  
Cormac G. M. Gahan ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Causative Agent ◽  
Draft Genome ◽  
Foodborne Pathogen ◽  
Genome Sequences ◽  
Serotype 1 ◽  
Serotype 4B

Listeria monocytogenes is a foodborne pathogen and the causative agent of listeriosis among humans and animals. The draft genome sequences of L. monocytogenes serotype 4b strains 944 and 2993 and serotype 1/2c strains 198 and 2932 are reported here.

scholarly journals Competition of Listeria monocytogenes Serotype 1/2a and 4b Strains in Mixed-Culture Biofilms

2009 ◽  
Vol 75 (18) ◽  
pp. 5846-5852 ◽  
Author(s):  
Youwen Pan ◽  
Frederick Breidt ◽  
Sophia Kathariou
Keyword(s):  
Listeria Monocytogenes ◽  
Mixed Culture ◽  
Processing System ◽  
Extracellular Dna ◽  
Microbial Competition ◽  
Growth And Survival ◽  
Specific Strain ◽  
Serotype 1 ◽  
Serotype 4B ◽  
Cell Densities

ABSTRACT The majority of Listeria monocytogenes isolates recovered from foods and the environment are strains of serogroup 1/2, especially serotypes 1/2a and 1/2b. However, serotype 4b strains cause the majority of human listeriosis outbreaks. Our investigation of L. monocytogenes biofilms used a simulated food-processing system that consisted of repeated cycles of growth, sanitation treatment, and starvation to determine the competitive fitness of strains of serotypes 1/2a and 4b in pure and mixed-culture biofilms. Selective enumeration of strains of a certain serotype in mixed-culture biofilms on stainless steel coupons was accomplished by using serotype-specific quantitative PCR and propidium monoazide treatment to prevent amplification of extracellular DNA or DNA from dead cells. The results showed that the serotype 1/2a strains tested were generally more efficient at forming biofilms and predominated in the mixed-culture biofilms. The growth and survival of strains of one serotype were not inhibited by strains of the other serotype in mixed-culture biofilms. However, we found that a cocktail of serotype 4b strains survived and grew significantly better in mixed-culture biofilms containing a specific strain of serotype 1/2a (strain SK1387), with final cell densities averaging 0.5 log10 CFU/cm2 higher than without the serotype 1/2a strain. The methodology used in this study contributed to our understanding of how environmental stresses and microbial competition influence the survival and growth of L. monocytogenes in pure and mixed-culture biofilms.

Behavior of Listeria monocytogenes during the Manufacture, Ripening, and Cold Storage of Afuega'l Pitu Cheese

1997 ◽  
Vol 60 (6) ◽  
pp. 689-693 ◽  
Author(s):  
ABELARDO MARGOLLES ◽  
ANA RODRÍGUEZ ◽  
CLARA G. de los REYES-GAVILÁN
Keyword(s):  
Listeria Monocytogenes ◽  
Lactococcus Lactis ◽  
Cell Damage ◽  
Salt Content ◽  
Northern Spain ◽  
Growth And Survival ◽  
Lactococcus Lactis Subsp ◽  
Whole Milk ◽  
Serotype 1 ◽  
Serotype 4B

Afuega'l Pitu is an artisanal acid-coagulated cheese manufactured in Asturias (northern Spain) and mainly consumed between the 3rd and the 30th day of ripening. Six cheese-making trials were performed in a pilot plant by using pasteurized whole milk inoculated with Listeria monocytogenes (strain L2 [serotype 1/2a], L39, or L41 [serotype 4b]) to ca. 2.7 log CFU/ml. A starter containing three strains, Lactococcus lactis subsp. lactis IPLA 947, Lactococcus lactis subsp. lactis biovar diacetylactis IPLA 838, and Leuconostoc citreum IPLA 616, grown separately in milk and combined in the volumetric proportion 3:1:1.3 was used. During the acidification L. monocytogenes counts increased 2.78- to 7.03-fold, depending on the strain, and remained within the curd; from this time counts decreased abruptly and were not detected in cheeses beyond the 7th day. The average pH in the curd was 4.43, and it decreased to around 4.0 in 5- to 7-day-old cheeses. These pH values were near the tolerance limit for L. monocytogenes and probably caused cell damage. Although moisture, aw, and NaCl levels were not limiting for the growth and survival of L. monocytogenes, salt content must be considered as a contributing factor in L. monocytogenes inactivation. Finally, the L2 strain grew better in curd and was slightly more resistant to low pH and refrigeration than strain L39 or L41. The manufacture of Afuega'l Pitu cheese from pasteurized milk and the design of a specific starter from the autochthonous lactic microbiota can lead to a safer product that can be consumed after very short ripening periods.

scholarly journals Molecular characterization of Listeria monocytogenes isolated from animal products in a city of Northern Brazil

2013 ◽  
Vol 43 (8) ◽  
pp. 1443-1448 ◽  
Author(s):  
Lilyan Rosmery Luizaga de Monteiro ◽  
Albenones José de Mesquita ◽  
Maria Cláudia Dantas Porfirio Borges André ◽  
Juliana Lamaro Cardoso
Keyword(s):  
Listeria Monocytogenes ◽  
Foodborne Pathogen ◽  
Meat Products ◽  
Ground Beef ◽  
Restriction Enzymes ◽  
Common Source ◽  
Animal Products ◽  
Public Markets ◽  
Serotype 1 ◽  
Serotype 4B

Listeria monocytogenes, a foodborne pathogen causes listeriosis, a fatal disease in about 30% of cases that affects mainly immunocompromised persons. The aim of this research was to characterize L. monocytogenes pulsed-field gel electrophoresis (PFGE) types isolated from meat products collected at public markets in Araguaina city, TO. Sixty samples of raw ground beef and frescal sausage were analyzed during the second half of 2008. Five out of 30 samples (16.7%) of raw ground beef tested positive for L. monocytogenes, three of which were classified as serotype 1/2b and two as serotype 4b. Among the 30 samples of sausage collected, two strains of L. monocytogenes were isolated (6.7%), one of them belonging to serotype 1/2a and the other belonging to serotype 1/2b. The restriction enzymes used were ApaI and SmaI. Similarities among the strains were determined by Dice coefficient. The macro restriction profile obtained by using SmaI enzyme allowed the distribution of seven strains in two clusters, two pulsotypes and two subtypes. The result indicates that L. monocytogenes isolates, belonging to serotype 4b, 1/2a and 1/2b, are strongly correlated within the same serotype group, and in some cases among different serotypes, suggesting that they have a common source.

Gene Transcription Patterns of pH- and Salt-Stressed Listeria monocytogenes Cells in Simulated Gastric and Pancreatic Conditions

2014 ◽  
Vol 77 (2) ◽  
pp. 254-261 ◽  
Author(s):  
MARIOS MATARAGAS ◽  
ANNA GREPPI ◽  
KALLIOPI RANTSIOU ◽  
LUCA COCOLIN
Keyword(s):  
Life Cycle ◽  
Listeria Monocytogenes ◽  
Reference Strain ◽  
Expression Patterns ◽  
Wild Strain ◽  
Hostile Environment ◽  
Fermented Sausage ◽  
Laboratory Reference ◽  
Serotype 1 ◽  
Serotype 4B

A Listeria monocytogenes subgenomic array, targeting 54 genes involved in the adhesion, adaptation, intracellular life cycle, invasion, and regulation of the infection cycle was used to investigate the gene expression patterns of acid- and salt-stressed Listeria cells after exposure to conditions similar to those in gastric and pancreatic fluids. Three L. monocytogenes strains, one laboratory reference strain (EGDe) and two food isolates (wild strain 12 isolated from milk and wild strain 3 isolated from fermented sausage), were used during the studies. Differences in the expressed genes were observed between the gastric and pancreatic treatments and also between the serotypes. Increased transcripts were observed of the genes belonging to the adaptation and regulation group for serotype 4b (strain 12) and to the invasion and regulation group for serotype 1/2a (strain EGDe). Interestingly, no significantly differentially expressed genes were found for serotype 3c (strain 3) in most cases. The genes related to adaptation (serotype 1/2a) and to intracellular life cycle and invasion (serotype 4b) were down-regulated in order to cope with the hostile environment of the gastric and pancreatic fluids. These findings may provide experimental evidence for the dominance of serotypes 1/2a and 4b in clinical cases of listeriosis and for the sporadic occurrence of serotype 3c.

scholarly journals Construction, Characterization, and Use of Two Listeria monocytogenes Site-Specific Phage Integration Vectors

2002 ◽  
Vol 184 (15) ◽  
pp. 4177-4186 ◽  
Author(s):  
Peter Lauer ◽  
Man Yin Nora Chow ◽  
Martin J. Loessner ◽  
Daniel A. Portnoy ◽  
Richard Calendar
Keyword(s):  
Listeria Monocytogenes ◽  
Lethal Dose ◽  
Attachment Site ◽  
Donor Cell ◽  
Single Copy ◽  
Cell Extract ◽  
Site Specific ◽  
Specific Phage ◽  
Serotype 1 ◽  
Serotype 4B

ABSTRACT Two site-specific shuttle integration vectors were developed with two different chromosomal bacteriophage integration sites to facilitate strain construction in Listeria monocytogenes. The first vector, pPL1, utilizes the listeriophage U153 integrase and attachment site within the comK gene for chromosomal insertion. pPL1 contains a useful polylinker, can be directly conjugated from Escherichia coli into L. monocytogenes, forms stable, single-copy integrants at a frequency of ∼10−4 per donor cell, and can be used in the L. monocytogenes 1/2 and 4b serogroups. Methods for curing endogenous prophages from the comK attachment site in 10403S-derived strains were developed. pPL1 was used to introduce the hly and actA genes at comK-attBB′ in deletion strains derived from 10403S and SLCC-5764. These strains were tested for second-site complementation in hemolysin assays, plaquing assays, and cell extract motility assays. Unlike plasmid-complemented strains, integrated pPL1-complemented strains were fully virulent in the mouse 50% lethal dose assay. Additionally, the PSA phage attachment site on the L. monocytogenes chromosome was characterized, and pPL1 was modified to integrate at this site. The listeriophage PSA integrates in the 3′ end of an arginine tRNA gene. There are 17 bp of DNA identity between the bacterial and phage attachment sites. The PSA prophage DNA sequence reconstitutes a complete tRNAArg gene. The modified vector, pPL2, was integration proficient at the same frequency as pPL1 in common laboratory serotype 1/2 strains as well as serotype 4b strains.

scholarly journals Conservation of Genomic Localization and Sequence Content of Sau3AI-Like Restriction-Modification Gene Cassettes among Listeria monocytogenes Epidemic Clone I and Selected Strains of Serotype 1/2a

2010 ◽  
Vol 76 (16) ◽  
pp. 5577-5584 ◽  
Author(s):  
Suleyman Yildirim ◽  
Driss Elhanafi ◽  
Wen Lin ◽  
Anthony D. Hitchins ◽  
Robin M. Siletzky ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gene Cassette ◽  
Conclusive Evidence ◽  
Gene Encoding ◽  
Epidemic Clone ◽  
Food Borne ◽  
Serotype 1 ◽  
Serotype 4B ◽  
Clonal Population Structure ◽  
Restriction Modification

ABSTRACTListeria monocytogenesis a food-borne pathogen with a clonal population structure and apparently limited gene flow between strains of different lineages. Strains of epidemic clone I (ECI) have been responsible for numerous outbreaks and invariably have DNA that is resistant to digestion by Sau3AI, suggesting methylation of cytosine at GATC sites. A putative restriction-modification (RM) gene cassette has been identified in the genome of the ECI strain F2365 and all other tested ECI strains but is absent from other strains of the same serotype (4b). Homologous RM cassettes have not been reported amongL. monocytogenesisolates of other serotypes. Furthermore, conclusive evidence for the involvement of this RM cassette in the Sau3AI resistance phenotype of ECI strains has been lacking. In this study, we describe a highly conserved RM cassette in certain strains of serotypes 1/2a and 4a that have Sau3AI-resistant DNA. In these strains the RM cassette was in the same genomic location as in the ECI reference strain F2365. The cassette included a gene encoding a putative recombinase, suggesting insertion via site-specific recombination. Deletion of the RM cassette in the ECI strain F2365 and the serotype 1/2a strain A7 rendered the DNA of both strains susceptible to Sau3AI digestion, providing conclusive evidence that the cassette includes a gene required for methylation of cytosine at GATC sites in both strains. The findings suggest that, in addition to its presence in ECI strains, this RM cassette and the accompanying genomic DNA methylation is also encountered among selected strains of other lineages.

scholarly journals Comparative Proteomic Analysis of Listeria monocytogenes Strains F2365 and EGD

2008 ◽  
Vol 75 (2) ◽  
pp. 366-373 ◽  
Author(s):  
Janet R. Donaldson ◽  
Bindu Nanduri ◽  
Shane C. Burgess ◽  
Mark L. Lawrence
Keyword(s):  
Listeria Monocytogenes ◽  
Protein Identification ◽  
Altered Expression ◽  
Genetic Lineages ◽  
Multidimensional Protein Identification Technology ◽  
Food Borne ◽  
Flagellar Biosynthesis ◽  
Serotype 1 ◽  
Serotype 4B ◽  
Biological Differences

ABSTRACT Listeria monocytogenes is a gram-positive, food-borne pathogen that causes disease in both humans and animals. There are three major genetic lineages of L. monocytogenes and 13 serovars. To further our understanding of the differences that exist between different genetic lineages/serovars of L. monocytogenes, we analyzed the global protein expression of the serotype 1/2a strain EGD and the serotype 4b strain F2365 during early-stationary-phase growth at 37°C. Using multidimensional protein identification technology with electrospray ionization tandem mass spectrometry, we identified 1,754 proteins from EGD and 1,427 proteins from F2365, of which 1,077 were common to both. Analysis of proteins that had significantly altered expression between strains revealed potential biological differences between these two L. monocytogenes strains. In particular, the strains differed in expression of proteins involved in cell wall physiology and flagellar biosynthesis, as well as DNA repair proteins and stress response proteins.

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