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.

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 Ability of the Listeria monocytogenes Strain Scott A To Cause Systemic Infection in Mice Infected by the Intragastric Route

2002 ◽  
Vol 68 (6) ◽  
pp. 2893-2900 ◽  
Author(s):  
Charles J. Czuprynski ◽  
Nancy G. Faith ◽  
Howard Steinberg
Keyword(s):  
Listeria Monocytogenes ◽  
Systemic Infection ◽  
Laboratory Strain ◽  
Gastric Fluid ◽  
Virulence Determinants ◽  
Food Borne ◽  
Serotype 1 ◽  
Serotype 4B ◽  
Prior Administration

ABSTRACT Listeriosis is an important food-borne disease that causes high rates of morbidity and mortality. For reasons that are not clear, most large outbreaks of human listeriosis involve Listeria monocytogenes serotype 4b. Relatively little is known about the pathogenesis of listeriosis following gastrointestinal exposure to food-borne disease isolates of L. monocytogenes. In the present study, we investigated the pathogenesis of systemic infection by the food-borne isolate Scott A in an intragastric (i.g.) mouse challenge model. We found that the severity of infection with L. monocytogenes Scott A was increased in mice made neutropenic by administration of monoclonal antibody RB6-8C5. This observation was similar to a previous report on a study with the laboratory strain L. monocytogenes EGD. Prior administration of sodium bicarbonate did not enhance the virulence of L. monocytogenes strain Scott A for i.g. inoculated mice. Following i.g. inoculation of mice, two serotype 4b strains of L. monocytogenes (Scott A and 101M) achieved a greater bacterial burden in the spleen and liver and elicited more severe histopathological damage to those organs than did a serotype 1/2a strain (EGD) and a serotype 1/2b stain (CM). Of the four strains tested, only strain CM exhibited poor survival in synthetic gastric fluid in vitro. The other three strains exhibited similar patterns of survival at pHs of greater than 5 and relatively rapid (<30 min) loss of viability at pHs of less than 5.0. Growth of L. monocytogenes Scott A at temperatures of 12.5 to 37°C did not affect its ability to cause systemic infection in i.g. inoculated mice. These observations suggest that the serotype 4b L. monocytogenes strains Scott A and 101M possess one or more virulence determinants that make them better able to cause systemic infection following inoculation via the g.i. tract than do the serotype 1/2 strains EGD and CM.

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 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 Synergistic Effects of Sodium Chloride, Glucose, and Temperature on Biofilm Formation by Listeria monocytogenes Serotype 1/2a and 4b Strains

2010 ◽  
Vol 76 (5) ◽  
pp. 1433-1441 ◽  
Author(s):  
Youwen Pan ◽  
Frederick Breidt ◽  
Lisa Gorski
Keyword(s):  
Growth Rate ◽  
Listeria Monocytogenes ◽  
Sodium Chloride ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Synergistic Effects ◽  
Matrix Material ◽  
Serotype 1 ◽  
Serotype 4B

ABSTRACT Biofilm formation by Listeria monocytogenes is generally associated with its persistence in the food-processing environment. Serotype 1/2a strains make up more than 50% of the total isolates recovered from food and the environment, while serotype 4b strains are most often associated with major outbreaks of human listeriosis. Using a microplate assay with crystal violet staining, we examined biofilm formation by 18 strains of each serotype in tryptic soy broth with varying concentrations of glucose (from 0.25% to 10.0%, wt/vol), sodium chloride (from 0.5% to 7.0%, wt/vol) and ethanol (from 1% to 5.0%, vol/vol), and at different temperatures (22.5�C, 30�C, and 37�C). A synergistic effect on biofilm formation was observed for glucose, sodium chloride, and temperature. The serotype 1/2a strains generally formed higher-density biofilms than the 4b strains under most conditions tested. Interestingly, most serotype 4b strains had a higher growth rate than the 1/2a strains, suggesting that the growth rate may not be directly related to the capacity for biofilm formation. Crystal violet was found to stain both bacterial cells and biofilm matrix material. The enhancement in biofilm formation by environmental factors was apparently due to the production of extracellular polymeric substances instead of the accumulation of viable biofilm cells.

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 Genetic Diversity of Listeria monocytogenes Strains from a High-Prevalence Dairy Farm

2005 ◽  
Vol 71 (10) ◽  
pp. 5893-5899 ◽  
Author(s):  
Monica K. Borucki ◽  
Clive C. Gay ◽  
James Reynolds ◽  
Katherine L. McElwain ◽  
So Hyun Kim ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Pacific Northwest ◽  
Dairy Farm ◽  
Clinical Strain ◽  
Time Points ◽  
Source Of Infection ◽  
Food Borne ◽  
High Prevalence ◽  
Serotype 1 ◽  
On Farm

ABSTRACT Listeria monocytogenes is a significant food-borne human and veterinary pathogen. Contaminated silage commonly leads to disease in livestock, but the pervasive nature of the bacterium can make it difficult to identify the source of infection. An investigation of bovine listeriosis that occurred on a Pacific Northwest dairy farm (“farm A”) revealed that the clinical strain was closely related to fecal strains from asymptomatic cows, and that farm environment was heavily contaminated with a diversity of L. monocytogenes strains. In addition, the farm A clinical strain was closely related to clinical and environmental strains obtained 1 year prior from a second Northwest dairy farm (“farm B”). To investigate the source(s) of contamination on farm A, environmental samples were collected from farm A at two time points. Pulsed-field gel electrophoresis characterization of 538 isolates obtained from that farm identified 57 different AscI pulsovars. Fecal isolates obtained from individual cows were the most genetically diverse, with up to 94% of fecal samples containing more than one pulsovar. The maximum numbers of pulsovars and serotypes isolated from a fecal sample of one cow were 6 and 4, respectively. Serotype 1/2a was isolated most frequently at both time points. Microarray genotyping of bovine listeriosis, fecal, and silage strains from both farms identified four probes that differentiated listeriosis strains from environmental strains; however, no probe was common to both bovine listeriosis strains.

scholarly journals prfA-Like Transcription Factor Genelmo0753Contributes to l-Rhamnose Utilization in Listeria monocytogenes Strains Associated with Human Food-Borne Infections

2013 ◽  
Vol 79 (18) ◽  
pp. 5584-5592 ◽  
Author(s):  
Joelle K. Salazar ◽  
Zhuchun Wu ◽  
P. David McMullen ◽  
Qin Luo ◽  
Nancy E. Freitag ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Listeria Monocytogenes ◽  
Bacterial Pathogen ◽  
Phenol Red ◽  
Flagellar Motility ◽  
Content Type ◽  
Genetic Lineages ◽  
Functional Roles ◽  
Food Borne ◽  
Virulence Regulator

ABSTRACTListeria monocytogenesis a food-borne bacterial pathogen and the causative agent of human and animal listeriosis. Among the three major genetic lineages ofL. monocytogenes(i.e., LI, LII, and LIII), LI and LII are predominantly associated with food-borne listeriosis outbreaks, whereas LIII is rarely implicated in human infections. In a previous study, we identified a Crp/Fnr family transcription factor gene,lmo0753, that was highly specific to outbreak-associated LI and LII but absent from LIII. Lmo0753 shares two conserved functional domains, including a DNA binding domain, with the well-characterized master virulence regulator PrfA inL. monocytogenes. In this study, we constructedlmo0753deletion and complementation mutants in two fully sequencedL. monocytogenesLII strains, 10403S and EGDe, and compared the flagellar motility, phospholipase C production, hemolysis, and intracellular growth of the mutants and their respective wild types. Our results suggested thatlmo0753plays a role in hemolytic activity in both EGDe and 10403S. More interestingly, we found that deletion oflmo0753led to the loss ofl-rhamnose utilization in EGDe, but not in 10403S. RNA-seq analysis of EGDe Δ0753incubated in phenol red medium containingl-rhamnose as the sole carbon source revealed that 126 (4.5%) and 546 (19.5%) out of 2,798 genes in the EGDe genome were up- and downregulated more than 2-fold, respectively, compared to the wild-type strain. Genes related to biotin biosynthesis, general stress response, and rhamnose metabolism were shown to be differentially regulated. Findings from this study collectively suggested varied functional roles oflmo0753in different LIIL. monocytogenesstrain backgrounds associated with human listeriosis outbreaks.

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