scholarly journals Identification of Listeria monocytogenes Genes Involved in Salt and Alkaline-pH Tolerance

2003 ◽  
Vol 69 (6) ◽  
pp. 3137-3143 ◽  
Author(s):  
Rozenn Gardan ◽  
Pascale Cossart ◽  
Jean Labadie
Keyword(s):  
Salt Stress ◽  
Listeria Monocytogenes ◽  
Food Processing ◽  
Food Preservation ◽  
Alkaline Ph ◽  
Sequence Comparisons ◽  
Insertional Mutants ◽  
Ph Tolerance ◽  
Alkaline Tolerance ◽  
Intergenic Regions

ABSTRACT The capacity of Listeria monocytogenes to tolerate salt and alkaline stresses is of particular importance, as this pathogen is often exposed to such environments during food processing and food preservation. We screened a library of Tn917-lacZ insertional mutants in order to identify genes involved in salt and/or alkaline tolerance. We isolated six mutants sensitive to salt stress and 12 mutants sensitive to salt and alkaline stresses. The position of the insertion of the transposon was located in 15 of these mutants. In six mutants the transposon was inserted in intergenic regions, and in nine mutants it was inserted in genes. Most of the genes have unknown functions, but sequence comparisons indicated that they encode putative transporters.

scholarly journals Salt Stress Proteins Induced in Listeria monocytogenes

2002 ◽  
Vol 68 (4) ◽  
pp. 1491-1498 ◽  
Author(s):  
Ophélie Duché ◽  
Frédéric Trémoulet ◽  
Philippe Glaser ◽  
Jean Labadie
Keyword(s):  
Salt Stress ◽  
Listeria Monocytogenes ◽  
Food Processing ◽  
Stress Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Food Preservation ◽  
Terminal Sequence ◽  
Phosphotransferase System ◽  
General Metabolism ◽  
Survival Mechanisms

ABSTRACT The ability of Listeria monocytogenes to tolerate salt stress is of particular importance, as this pathogen is often exposed to such environments during both food processing and food preservation. In order to understand the survival mechanisms of L. monocytogenes, an initial approach using two-dimensional polyacrylamide gel electrophoresis was performed to analyze the pattern of protein synthesis in response to salt stress. Of 400 to 500 visible proteins, the synthesis of 40 proteins (P < 0.05) was repressed or induced at a higher rate during salt stress. Some of the proteins were identified on the basis of mass spectrometry or N-terminal sequence analysis and database searching. Twelve proteins showing high induction after salt stress were similar to general stress proteins (Ctc and DnaK), transporters (GbuA and mannose-specific phosphotransferase system enzyme IIAB), and general metabolism proteins (alanine dehydrogenase, CcpA, CysK, EF-Tu, Gap, GuaB, PdhA, and PdhD).

Role of Sigma B Factor in the Alkaline Tolerance Response of Listeria monocytogenes 10403S and Cross-Protection against Subsequent Ethanol and Osmotic Stress

2008 ◽  
Vol 71 (7) ◽  
pp. 1481-1485 ◽  
Author(s):  
EFSTATHIOS S. GIOTIS ◽  
MUDCHAREE JULOTOK ◽  
BRIAN J. WILKINSON ◽  
IAN S. BLAIR ◽  
DAVID A. MCDOWELL
Keyword(s):  
Listeria Monocytogenes ◽  
Osmotic Stress ◽  
Surface Cleaning ◽  
Cross Protection ◽  
Ethanol Stress ◽  
Alkaline Ph ◽  
Alkaline Tolerance ◽  
Wide Range ◽  
Type Strains

Many of the considerable abilities of Listeria monocytogenes to persist and grow in a wide range of adverse environmental conditions are thought to be at least partly under the control of the alternative sigma factor (σB), encoded by the sigB gene. However, little is known about the role of this master regulon in the impressive ability of Listeria to persist and grow under conditions of alkaline pH. In this study, Northern blot analysis of parent Listeria mRNA revealed that alkali adaptation (pH 9.5 for 1 h) significantly increased the expression of sigB-derived mRNA. The study included a comparison of the relative survival of mid-exponential populations of adapted and nonadapted parent type (σB expressing) and mutant (not σB expressing, ΔsigB) Listeria strains during subsequent alkaline (pH 12.0), osmotic (25% NaCl, wt/vol), or ethanol (16.5%) stress. Alkali-adapted parent strains were more resistant to pH 12.0 than were adapted ΔsigB type strains, but both alkali-adapted parent and ΔsigB strains were more resistant to pH 12.0 than were nonadapted strains. Alkali-adapted parent strains were more resistant to osmotic stress than were adapted ΔsigB type strains. No significant differences in viability were observed between alkali-adapted parent and ΔsigB strains after ethanol stress, suggesting that cross-protection against osmotic stress is mediated by σB whereas cross-protection against ethanol is σB independent. Overall, alkali-induced cross-protection against osmotic and ethanol challenges may have serious implications for food safety and human health because such stress conditions are routinely used as part of food preservation and surface cleaning processes.

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.

Listeria monocytogenes strains encoding premature stop codons in inlA invade mice and guinea pig fetuses in orally dosed dams

2013 ◽  
Vol 62 (12) ◽  
pp. 1799-1806 ◽  
Author(s):  
Anne Holch ◽  
Hanne Ingmer ◽  
Tine Rask Licht ◽  
Lone Gram
Keyword(s):  
Listeria Monocytogenes ◽  
Guinea Pig ◽  
Food Processing ◽  
Guinea Pigs ◽  
Stop Codon ◽  
Fetal Liver ◽  
Fatal Case ◽  
Premature Stop Codon ◽  
Tissue Samples ◽  
Pregnant Mice

Listeria monocytogenes is an important food-borne bacterial pathogen and listeriosis can result in abortions in pregnant women. The bacterium can colonize food-processing environments, where specific molecular subtypes can persist for years. The purpose of this study was to determine the virulence potential of a group of food-processing persistent L. monocytogenes strains encoding a premature stop codon in inlA (encoding internalin A) by using two orally dosed models, pregnant mice and pregnant guinea pigs. A food-processing persistent strain of L. monocytogenes invaded placentas (n = 58; 10 % positive) and fetuses (3 % positive) of pregnant mice (n = 9 animals per strain), similar to a genetically manipulated murinized strain, EGD-e InlA m* (n = 61; 3 and 2 %, respectively). In pregnant guinea pigs (n = 9 animals per bacterial strain), a maternofetal strain (from a human fetal clinical fatal case) was isolated from 34 % of placenta samples (n = 50), whereas both food-processing persistent strains were found in 5 % of placenta samples (n = 36 or 37). One of the food-processing persistent strains, N53-1, was found in up to 8 % of guinea pig fetal liver and brain samples, whereas the maternofetal control was found in 6 % of fetal tissue samples. As the food-processing persistent strains carry a premature stop codon in inlA but are invasive in orally dosed pregnant mice and guinea pigs, we hypothesize that listerial crossing of the placental barrier can occur by a mechanism that is independent of an interaction between E-cadherin and InlA.

scholarly journals Genome Sequencing Identifies Two Nearly Unchanged Strains of Persistent Listeria monocytogenes Isolated at Two Different Fish Processing Plants Sampled 6 Years Apart

2013 ◽  
Vol 79 (9) ◽  
pp. 2944-2951 ◽  
Author(s):  
Anne Holch ◽  
Kristen Webb ◽  
Oksana Lukjancenko ◽  
David Ussery ◽  
Benjamin M. Rosenthal ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Random Amplified Polymorphic Dna ◽  
The United States ◽  
Genome Comparison ◽  
Physiological Factors ◽  
Content Type ◽  
Human Pathogenic Bacterium ◽  
A Genome ◽  
Genome Analyses

ABSTRACTListeria monocytogenesis a food-borne human-pathogenic bacterium that can cause infections with a high mortality rate. It has a remarkable ability to persist in food processing facilities. Here we report the genome sequences for twoL. monocytogenesstrains (N53-1 and La111) that were isolated 6 years apart from two different Danish fish processers. Both strains are of serotype 1/2a and belong to a highly persistent DNA subtype (random amplified polymorphic DNA [RAPD] type 9). We demonstrate usingin silicoanalyses that both strains belong to the multilocus sequence typing (MLST) type ST121 that has been isolated as a persistent subtype in several European countries. The purpose of this study was to use genome analyses to identify genes or proteins that could contribute to persistence. In a genome comparison, the two persistent strains were extremely similar and collectively differed from the reference lineage II strain, EGD-e. Also, they differed markedly from a lineage I strain (F2365). On the proteome level, the two strains were almost identical, with a predicted protein homology of 99.94%, differing at only 2 proteins. No single-nucleotide polymorphism (SNP) differences were seen between the two strains; in contrast, N53-1 and La111 differed from the EGD-e reference strain by 3,942 and 3,471 SNPs, respectively. We included a persistentL. monocytogenesstrain from the United States (F6854) in our comparisons. Compared to nonpersistent strains, all three persistent strains were distinguished by two genome deletions: one, of 2,472 bp, typically contains the gene forinlF, and the other, of 3,017 bp, includes three genes potentially related to bacteriocin production and transport (lmo2774,lmo2775, and the 3′-terminal part oflmo2776). Further studies of highly persistent strains are required to determine if the absence of these genes promotes persistence. While the genome comparison did not point to a clear physiological explanation of the persistent phenotype, the remarkable similarity between the two strains indicates that subtypes with specific traits are selected for in the food processing environment and that particular genetic and physiological factors are responsible for the persistent phenotype.

scholarly journals Sterol-response pathways mediate alkaline survival in diverse fungi

2020 ◽  
Author(s):  
Hannah E. Brown ◽  
Calla L. Telzrow ◽  
Joseph W. Saelens ◽  
Larissa Fernandes ◽  
J. Andrew Alspaugh
Keyword(s):  
Fungal Pathogens ◽  
Fungal Infections ◽  
Membrane Integrity ◽  
Microbial Pathogens ◽  
Alkaline Ph ◽  
Ph Response ◽  
Ph Tolerance ◽  
Alkaline Conditions ◽  
Host Infection ◽  
The Impact

AbstractThe ability for cells to maintain homeostasis in the presence of extracellular stress is essential for their survival. Stress adaptations are especially important for microbial pathogens to respond to rapidly changing conditions, such as those encountered during the transition from the environment to the infected host. Many fungal pathogens have acquired the ability to quickly adapt to changes in extracellular pH to promote their survival in the various micro-environments encountered during a host infection. For example, the fungal-specific Rim/Pal alkaline response pathway has been well characterized in many fungal pathogens, including Cryptococcus neoformans. However, alternative mechanisms for sensing and responding to host pH have yet to be extensively studied. Recent observations from a genetic screen suggest that the C. neoformans sterol homeostasis pathway is required for growth at elevated pH. This work explores interactions among mechanisms of membrane homeostasis, alkaline pH tolerance, and Rim pathway activation. We find that the sterol homeostasis pathway is necessary for growth in an alkaline environment, and that an elevated pH is sufficient to induce Sre1 activation. This pH-mediated activation of the Sre1 transcription factor is linked to the biosynthesis of ergosterol, but is not dependent on Rim pathway signaling, suggesting that these two pathways are responding to alkaline pH independently. Furthermore, we discover that C. neoformans is more susceptible to membrane-targeting antifungals in alkaline conditions highlighting the impact of micro-environmental pH on the treatment of invasive fungal infections. Together, these findings further connect membrane integrity and composition with the fungal pH response and pathogenesis.

scholarly journals Sublethal Injury Caused to Listeria monocytogenes by Natural Plant Extracts: Case Study on Grape Seed Extract and Garlic Extract

2019 ◽  
Vol 9 (13) ◽  
pp. 2731 ◽  
Author(s):  
Davy Verheyen ◽  
Maria Baka ◽  
Jan F. M. Van Impe
Keyword(s):  
Listeria Monocytogenes ◽  
Plant Extracts ◽  
Membrane Damage ◽  
Grape Seed Extract ◽  
Grape Seed ◽  
Seed Extract ◽  
Food Preservation ◽  
Natural Plant ◽  
Novel Food ◽  
Sublethal Injury

Natural antimicrobials, such as grape seed extract (GSE) and garlic extract (GE), are often used as (a part of) novel food preservation technologies, especially due to their antilisterial effect. However, little is known on the extent of sublethal injury (SI) these extracts cause to Listeria monocytogenes, possibly leading to overestimated efficacies for such novel technologies. The influence of both extracts on the sublethal injury of L. monocytogenes strain LMG23775 was investigated, also using propidium iodide staining to investigate the nature of the injury. Minimum inhibitory concentrations were 500 mg gallic acid equivalents (GAE)/L and 7.5 μg allicin/mL for GSE and GE, respectively. These concentrations slowed down the growth of L. monocytogenes, while only causing a significant amount of SI for GSE. Pure extracts caused inactivation of the cells, with GSE being the most effective. Sublethal injury from pure GSE was mainly caused by membrane damage. In pure GE, a significant amount of SI, not caused by membrane damage, was also present, albeit less pronounced than in pure GSE. In conclusion, both extracts cause a significant amount of sublethal injury to L. monocytogenes, which is not taken into account in relevant studies investigating novel food preservation applications involving natural plant extracts.

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