Evaluation of cold growth and related gene transcription responses associated with Listeria monocytogenes strains of different origins

Arguedas-Villa, Carolina; Stephan, Roger; Tasara, Taurai

doi:10.1016/j.fm.2010.02.009

Cited by 38 publications

(38 citation statements)

References 52 publications

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“…Scatter plots show mean values with standard deviations. groups, similar to observations previously made among L. monocytogenes strains derived from different origins (36). When sample origin was examined, the majority of fast-adapting strains were recovered from RTE foods, although differences were not significant.…”

Section: Discussionsupporting

confidence: 85%

“…A subset of isolates (n ϭ 33) representing L. monocytogenes strains with full-length inlA, a 3-codon deletion (amino acids [aa] 738 to 740), and each type of PMSC observed in our collection was assessed for cold growth adaptation as described by Arguedas-Villa et al (36). In short, a single colony was inoculated into 10 ml brain heart infusion broth (Oxoid, Ottawa, ON) and grown overnight at 37°C with shaking (220 rpm) (ϳ10 9 CFU/ml).…”

Section: Bacterial Isolatesmentioning

confidence: 99%

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Examination of Food Chain-Derived Listeria monocytogenes Strains of Different Serotypes Reveals Considerable Diversity in inlA Genotypes, Mutability, and Adaptation to Cold Temperatures

Kovačević

Arguedas-Villa

Woźniak

et al. 2013

Appl Environ Microbiol

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c Listeria monocytogenes strains belonging to serotypes 1/2a and 4b are frequently linked to listeriosis. While inlA mutations leading to premature stop codons (PMSCs) and attenuated virulence are common in 1/2a, they are rare in serotype 4b. We observed PMSCs in 35% of L. monocytogenes isolates (n ‫؍‬ 54) recovered from the British Columbia food supply, including serotypes 1/2a (30%), 1/2c (100%), and 3a (100%), and a 3-codon deletion (amino acid positions 738 to 740) seen in 57% of 4b isolates from fish-processing facilities. Caco-2 invasion assays showed that two isolates with the deletion were significantly more invasive than EGD-SmR (P < 0.0001) and were either as (FF19-1) or more (FE13-1) invasive than a clinical control strain (08-5578) (P ‫؍‬ 0.006). To examine whether serotype 1/2a was more likely to acquire mutations than other serotypes, strains were plated on agar with rifampin, revealing 4b isolates to be significantly more mutable than 1/2a, 1/2c, and 3a serotypes (P ‫؍‬ 0.0002). We also examined the ability of 33 strains to adapt to cold temperature following a downshift from 37°C to 4°C. Overall, three distinct coldadapting groups (CAG) were observed: 46% were fast (<70 h), 39% were intermediate (70 to 200 h), and 15% were slow (>200 h) adaptors. Intermediate CAG strains (70%) more frequently possessed inlA PMSCs than did fast (20%) and slow (10%) CAGs; in contrast, 87% of fast adaptors lacked inlA PMSCs. In conclusion, we report food chain-derived 1/2a and 4b serotypes with a 3-codon deletion possessing invasive behavior and the novel association of inlA genotypes encoding a full-length InlA with fast cold-adaptation phenotypes.

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Section: Discussionsupporting

confidence: 85%

Section: Bacterial Isolatesmentioning

confidence: 99%

Examination of Food Chain-Derived Listeria monocytogenes Strains of Different Serotypes Reveals Considerable Diversity in inlA Genotypes, Mutability, and Adaptation to Cold Temperatures

Kovačević

Arguedas-Villa

Woźniak

et al. 2013

Appl Environ Microbiol

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“…Antibody-based serotyping was conducted on a subset of isolates ( n = 91) within both the current study and previous studies (Arguedas-Villa et al, 2010; Kovacevic et al, 2013). Remaining isolates were assigned one or more possible serotypes by performing a MegaBLAST search (>95% nt identity) ncbi-blast+ v. 2.3.0 available at: ftp://ftp.ncbi.nlm.nih.gov/blast/executables/blast+/LATEST/) for four genes used in a multiplex PCR developed by Doumith et al (2004).…”

Section: Methodsmentioning

confidence: 99%

Genotypes Associated with Listeria monocytogenes Isolates Displaying Impaired or Enhanced Tolerances to Cold, Salt, Acid, or Desiccation Stress

et al. 2017

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The human pathogen Listeria monocytogenes is a large concern in the food industry where its continuous detection in food products has caused a string of recalls in North America and Europe. Most recognized for its ability to grow in foods during refrigerated storage, L. monocytogenes can also tolerate several other food-related stresses with some strains possessing higher levels of tolerances than others. The objective of this study was to use a combination of phenotypic analyses and whole genome sequencing to elucidate potential relationships between L. monocytogenes genotypes and food-related stress tolerance phenotypes. To accomplish this, 166 L. monocytogenes isolates were sequenced and evaluated for their ability to grow in cold (4°C), salt (6% NaCl, 25°C), and acid (pH 5, 25°C) stress conditions as well as survive desiccation (33% RH, 20°C). The results revealed that the stress tolerance of L. monocytogenes is associated with serotype, clonal complex (CC), full length inlA profiles, and the presence of a plasmid which was identified in 55% of isolates. Isolates with full length inlA exhibited significantly (p < 0.001) enhanced cold tolerance relative to those harboring a premature stop codon (PMSC) in this gene. Similarly, isolates possessing a plasmid demonstrated significantly (p = 0.013) enhanced acid tolerance. We also identified nine new L. monocytogenes sequence types, a new inlA PMSC, and several connections between CCs and the presence/absence or variations of specific genetic elements. A whole genome single-nucleotide-variants phylogeny revealed sporadic distribution of tolerant isolates and closely related sensitive and tolerant isolates, highlighting that minor genetic differences can influence the stress tolerance of L. monocytogenes. Specifically, a number of cold and desiccation sensitive isolates contained PMSCs in σB regulator genes (rsbS, rsbU, rsbV). Collectively, the results suggest that knowing the sequence type of an isolate in addition to screening for the presence of full-length inlA and a plasmid, could help food processors and food agency investigators determine why certain isolates might be persisting in a food processing environment. Additionally, increased sequencing of L. monocytogenes isolates in combination with stress tolerance profiling, will enhance the ability to identify genetic elements associated with higher risk strains.

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“…RNA profiles in samples were immediately preserved using the bacterial RNA Protect Reagent (Qiagen). Total RNA was extracted as previously described (Arguedas-Villa et al, 2010). Samples were controlled for potential residual DNA contamination prior to reverse transcription by amplification.…”

Section: Stress Exposure Total Rna Extraction and Cdna Synthesismentioning

confidence: 99%

“…Previously described amplification conditions were used and the 16S rRNA and lmo0501 gene specific primers were optimized to achieve specific target amplification ranging from 95 to 100% PCR efficiency (Arguedas-Villa et al, 2010). 7.2 ng of cDNA prepared as described above were used as templates in qPCR assays.…”

Section: Relative Quantification Of Lmo0501 Gene Expressionmentioning

confidence: 99%

The lmo0501 gene coding for a putative transcription activator protein in Listeria monocytogenes promotes growth under cold, osmotic and acid stress conditions

2011

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In L. monocytogenes EGDe, the lmo0501 gene locus encodes a protein similar to the mannitol transcription regulator (MltR) protein in B. subtilis and B. stearothermophilus. In this study we investigated its functional role in L. monocytogenes EGDe cells in view of growth under different stress conditions. Increased lmo0501 gene expression at mRNA level was detected in response to cold, osmotic and organic acid stress exposure. An EGDe Δlmo0501 mutant strain was diminished in growth compared to the wild type strain in minimal defined medium containing either glucose or fructose, as carbon sources. Furthermore the lmo0501 null mutant had retarded growth under cold (4°C), salt (NaCl) and organic acid stress conditions compared to the parental wild type strain. Our results confirm the role of the lmo0501 gene in view of adaptation of L. monocytogenes cells to stress conditions as well as a contribution to the efficient utilization of glucose and fructose as carbon sources.Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-60467 Originally published at: Michel, E. The lmo0501 gene coding for a putative transcription activator protein in Listeria monocytogenes promotes growth under cold, osmotic and acid stress conditions. 2011, University of Zurich, Vetsuisse Faculty.

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Evaluation of cold growth and related gene transcription responses associated with Listeria monocytogenes strains of different origins

Cited by 38 publications

References 52 publications

Examination of Food Chain-Derived Listeria monocytogenes Strains of Different Serotypes Reveals Considerable Diversity in inlA Genotypes, Mutability, and Adaptation to Cold Temperatures

Examination of Food Chain-Derived Listeria monocytogenes Strains of Different Serotypes Reveals Considerable Diversity in inlA Genotypes, Mutability, and Adaptation to Cold Temperatures

Genotypes Associated with Listeria monocytogenes Isolates Displaying Impaired or Enhanced Tolerances to Cold, Salt, Acid, or Desiccation Stress

The lmo0501 gene coding for a putative transcription activator protein in Listeria monocytogenes promotes growth under cold, osmotic and acid stress conditions

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