scholarly journals Comparative Analysis of Listeria monocytogenes Plasmids and Expression Levels of Plasmid-Encoded Genes during Growth under Salt and Acid Stress Conditions

Toxins ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 426 ◽  
Author(s):  
Patricia Hingston ◽  
Thomas Brenner ◽  
Lisbeth Truelstrup Hansen ◽  
Siyun Wang
Keyword(s):  
Listeria Monocytogenes ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Stress Conditions ◽  
Chaperone Protein ◽  
Nadh Peroxidase ◽  
Genes Encoding ◽  
Fold Induction ◽  
N 93 ◽  
Unique Plasmid

Listeria monocytogenes strains are known to harbour plasmids that confer resistance to sanitizers, heavy metals, and antibiotics; however, very little research has been conducted into how plasmids may influence L. monocytogenes’ ability to tolerate food-related stresses. To investigate this, a library (n = 93) of L. monocytogenes plasmid sequences were compared. Plasmid sequences were divided into two groups (G1 and G2) based on a repA phylogeny. Twenty-six unique plasmid types were observed, with 13 belonging to each of the two repA-based groups. G1 plasmids were significantly (p < 0.05) smaller than G2 plasmids but contained a larger diversity of genes. The most prevalent G1 plasmid (57,083 bp) was observed in 26 strains from both Switzerland and Canada and a variety of serotypes. Quantitative PCR (qPCR) revealed a >2-fold induction of plasmid-contained genes encoding an NADH peroxidase, cadmium ATPase, multicopper oxidase, and a ClpL chaperone protein during growth under salt (6% NaCl) and acid conditions (pH 5) and ProW, an osmolyte transporter, under salt stress conditions. No differences in salt and acid tolerance were observed between plasmid-cured and wildtype strains. This work highlights the abundance of specific plasmid types among food-related L. monocytogenes strains, the unique characteristics of G1 and G2 plasmids, and the possible contributions of plasmids to L. monocytogenes tolerance to food-related stresses.

scholarly journals Identification and Inactivation of Genetic Loci Involved with Lactobacillus acidophilus Acid Tolerance

2004 ◽  
Vol 70 (9) ◽  
pp. 5315-5322 ◽  
Author(s):  
M. Andrea Azcarate-Peril ◽  
Eric Altermann ◽  
Rebecca L. Hoover-Fitzula ◽  
Raul J. Cano ◽  
Todd R. Klaenhammer
Keyword(s):  
Amino Acid ◽  
Lactobacillus Acidophilus ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Low Ph ◽  
Open Reading Frames ◽  
Amino Acid Permease ◽  
Physiological Limits ◽  
Insertional Inactivation ◽  
Genes Encoding

ABSTRACT Amino acid decarboxylation-antiporter reactions are one of the most important systems for maintaining intracellular pH between physiological limits under acid stress. We analyzed the Lactobacillus acidophilus NCFM complete genome sequence and selected four open reading frames with similarities to genes involved with decarboxylation reactions involved in acid tolerance in several microorganisms. Putative genes encoding an ornithine decarboxylase, an amino acid permease, a glutamate γ-aminobutyrate antiporter, and a transcriptional regulator were disrupted by insertional inactivation. The ability of L. acidophilus to survive low-pH conditions, such as those encountered in the stomach or fermented dairy foods, was investigated and compared to the abilities of early- and late-stationary-phase cells of the mutants by challenging them with a variety of acidic conditions. All of the integrants were more sensitive to low pH than the parental strain. Interestingly, each integrant also exhibited an adaptive acid response during logarithmic growth, indicating that multiple mechanisms are present and orchestrated in L. acidophilus in response to acid challenge.

Induced Acid-Tolerance Response Confers Limited Nisin Resistance on Listeria monocytogenes Scott A

1996 ◽  
Vol 59 (9) ◽  
pp. 1003-1006 ◽  
Author(s):  
AMECHI OKEREKE ◽  
STERLING S. THOMPSON
Keyword(s):  
Listeria Monocytogenes ◽  
Acid Stress ◽  
Brain Heart Infusion ◽  
Acid Tolerance ◽  
Acid Resistance ◽  
Cross Protection ◽  
C Cells ◽  
Acid Tolerance Response ◽  
Tolerance Response ◽  
Resistance Trait

The presence of an inducible acid-tolerance response (ATR) in Listeria monocytogenes Scott A was established. Protection of cells with induced ATR against nisin-mediated inhibition and stress was also evaluated. ATR was induced in L. monocytogenes Scott A by culturing in brain heart infusion (BHI) broth buffered to pH 5.4. The unadapted cells were grown at pH 7.2. Both acid-adapted and unadapted cells were challenged at pH 3.3 and 4.3 at 35°C. The acid-adapted cells were 150- to 7,500-fold more resistant to acid stress at pH 3.3 than unadapted cells. Both cells were equally resistant to acid stress at pH 4.3. The acid-adapted and unadapted cells were exposed to 0, 0.3, 0.6, 1.2 and 1.5 μg of nisin per ml of buffered BHI broth at pH 6.0 for 90 min at 35°C. Cells with the induced acid-resistance trait were slightly more resistant to nisin than the unadapted cells. In the presence of 1.5 μg of nisin per ml, 47% of the acid-adapted cells survived compared to 41% of the unadapted cells. In the range of nisin concentration included in this study, there was no significant (P &lt; 0.05) difference in the nisin resistance of adapted and unadapted cells. The data suggest that ATR induction confers very limited cross protection against nisin stress and kill.

scholarly journals Comparative Analysis of the σB-Dependent Stress Responses in Listeria monocytogenes and Listeria innocua Strains Exposed to Selected Stress Conditions

2007 ◽  
Vol 74 (1) ◽  
pp. 158-171 ◽  
Author(s):  
Sarita Raengpradub ◽  
Martin Wiedmann ◽  
Kathryn J. Boor
Keyword(s):  
Listeria Monocytogenes ◽  
Stress Response ◽  
Stationary Phase ◽  
Stress Responses ◽  
Bacterial Species ◽  
Acid Stress ◽  
Listeria Innocua ◽  
Transcript Levels ◽  
Genes Encoding ◽  
Flagellar Proteins

ABSTRACT The alternative sigma factor σB contributes to transcription of stress response and virulence genes in diverse gram-positive bacterial species. The composition and functions of the Listeria monocytogenes and Listeria innocua σB regulons were hypothesized to differ due to virulence differences between these closely related species. Transcript levels in stationary-phase cells and in cells exposed to salt stress were characterized by microarray analyses for both species. In L. monocytogenes, 168 genes were positively regulated by σB; 145 of these genes were preceded by a putative σB consensus promoter. In L. innocua, 64 genes were positively regulated by σB. σB contributed to acid stress survival in log-phase cells for both species but to survival in stationary-phase cells only for L. monocytogenes. In summary, (i) the L. monocytogenes σB regulon includes >140 genes that are both directly and positively regulated by σB, including genes encoding proteins with importance in stress response, virulence, transcriptional regulation, carbohydrate metabolism, and transport; (ii) a number of L. monocytogenes genes encoding flagellar proteins show higher transcript levels in the ΔsigB mutant, and both L. monocytogenes and L. innocua ΔsigB null mutants have increased motility compared to the respective isogenic parent strains, suggesting that σB affects motility and chemotaxis; and (iii) although L. monocytogenes and L. innocua differ in σB-dependent acid stress resistance and have species-specific σB-dependent genes, the L. monocytogenes and L. innocua σB regulons show considerable conservation, with a common set of at least 49 genes that are σB dependent in both species.

scholarly journals σB Activation under Environmental and Energy Stress Conditions in Listeria monocytogenes

2006 ◽  
Vol 72 (8) ◽  
pp. 5197-5203 ◽  
Author(s):  
Soraya Chaturongakul ◽  
Kathryn J. Boor
Keyword(s):  
Listeria Monocytogenes ◽  
Cumene Hydroperoxide ◽  
Stress Conditions ◽  
Wild Type ◽  
Regulation Of Expression ◽  
Transcript Levels ◽  
Energy Stress ◽  
Expression Studies ◽  
Genes Encoding ◽  
Gene Expression Studies

ABSTRACT To measure σB activation in Listeria monocytogenes under environmental or energy stress conditions, quantitative reverse transcriptase PCR (TaqMan) was used to determine the levels of transcripts for the σB-dependent opuCA and clpC genes in strains having null mutations in genes encoding regulator of sigma B proteins (rsbT and rsbV) and sigma B (sigB) and in the L. monocytogenes wild-type 10403S strain under different stress conditions. The ΔsigB, ΔrsbT, and ΔrsbV strains previously exhibited increased hemolytic activities compared to the hemolytic activity of the wild-type strain; therefore, transcript levels for hly were also determined. RsbT, RsbV, and σB were all required for opuCA expression during growth under carbon-limiting conditions or following exposure to pH 4.5, salt, ethanol, or the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP). Expression of clpC was RsbT, RsbV, and σB dependent in the presence of CCCP but not under the other conditions. hly expression was not RsbT, RsbV, or σB dependent in the presence of either CCCP or salt. opuCA transcript levels did not increase in the presence of rapidly lethal stresses (i.e., pH 2.5 or 13 mM cumene hydroperoxide) despite the enhanced survival of the wild type compared with the survival of the mutant strains under these conditions. These findings highlight the importance of complementing phenotypic characterizations with gene expression studies to identify direct and indirect effects of null mutations in regulatory genes, such as sigB. Overall, our data show that while σB activation occurs through a single pathway under both environmental and energy stress conditions, regulation of expression of some stress response and virulence genes in the σB regulon (e.g., clpC) appears to require networks involving multiple transcriptional regulators.

Modeling Survival of Listeria monocytogenes in the Traditional Greek Soft Cheese Katiki

2008 ◽  
Vol 71 (9) ◽  
pp. 1835-1845 ◽  
Author(s):  
MARIOS MATARAGAS ◽  
VIRGINIA STERGIOU ◽  
GEORGE-JOHN E. NYCHAS
Keyword(s):  
Listeria Monocytogenes ◽  
Survival Rate ◽  
Survival Rates ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Initial Population ◽  
Bacterial Cells ◽  
Mathematical Equations ◽  
Estimation And Control ◽  
And Control

In the present work, survival of Listeria monocytogenes in the traditional Greek soft, spreadable cheese Katiki was studied throughout the shelf life of the product. Samples of finished cheese were inoculated with a cocktail of five L. monocytogenes strains (ca. 6 log CFU g−1) and stored at 5, 10, 15, and 20°C. Acid-stress adaptation or cross-protection to the same stress was also investigated by inoculation of acid-adapted cells in the product. The results showed that pathogen survival was biphasic. Various mathematical equations (Geeraerd, Cerf, Albert-Mafart, Whiting, Zwietering, and Baranyi models) were fitted to the experimental data. A thorough statistical analysis was performed to choose the best model. The Geeraerd model was finally selected, and the results revealed no acid tolerance acquisition (no significant differences, P &gt; 0.05, in the survival rates of the non–acid-adapted and acid-adapted cells). Secondary modeling (second-order polynomial with a0 = 0.8453, a1 = −0.0743, and a2 = 0.0059) of the survival rate (of sensitive population), and other parameters that were similar at all temperatures (fraction of initial population in the major population = 99.98%, survival rate of resistant population = 0.10 day−1, and initial population = 6.29 log CFU g−1), showed that survival of the pathogen was temperature dependent with bacterial cells surviving for a longer period of time at lower temperatures. Finally, the developed predictive model was successfully validated at two independent temperatures (12 and 17°C). This study underlines the usefulness of predictive modeling as a tool for realistic estimation and control of L. monocytogenes risk in food products. Such data are also useful when conducting risk assessment studies.

scholarly journals Effect of Food Processing-Related Stresses on Acid Tolerance of Listeria monocytogenes

2003 ◽  
Vol 69 (12) ◽  
pp. 7514-7516 ◽  
Author(s):  
Konstantinos P. Koutsoumanis ◽  
Patricia A. Kendall ◽  
John N. Sofos
Keyword(s):  
Listeria Monocytogenes ◽  
Food Processing ◽  
Adaptive Response ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Acid Resistance ◽  
Acidic Ph ◽  
Temperature Stresses ◽  
Subsequent Exposure ◽  
Effect Of Food

ABSTRACT Stationary-phase cells of Listeria monocytogenes grown in glucose-free or glucose-containing media were exposed for 90 min to various stresses, including acid stress (pH 4.0 to 7.0), osmotic stress (10.5 to 20.5% NaCl), and various temperatures (−5 to 50°C), and were further exposed to pH 3.5. Exposure to a mildly acidic (pH 5.0 to 6.0) environment provided protection of the pathogen against acid upon subsequent exposure. This adaptive response, however, was found to be strongly dependent on other environmental conditions during the shock, such as temperature or the simultaneous presence of a second stress factor (NaCl). Growth of L. monocytogenes in the presence of glucose resulted in enhanced survival of the pathogen at pH 3.5. Sublethal stresses other than acidic stresses, i.e., osmotic, heat, and low-temperature stresses, did not affect the acid resistance of L. monocytogenes (P > 0.5). More-severe levels of these stresses, however, resulted in sensitization of the pathogen to acid.

Listeria monocytogenes ArcA contributes to acid tolerance

2013 ◽  
Vol 62 (6) ◽  
pp. 813-821 ◽  
Author(s):  
Changyong Cheng ◽  
Jianshun Chen ◽  
Ying Shan ◽  
Chun Fang ◽  
Yuan Liu ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Bacterial Load ◽  
Foodborne Pathogen ◽  
Acid Stress ◽  
Intestinal Barrier ◽  
Systemic Infection ◽  
Acid Tolerance ◽  
Gastric Fluid ◽  
Arginine Deiminase

The foodborne pathogen Listeria monocytogenes is able to colonize the human and animal intestinal tracts and subsequently crosses the intestinal barrier, causing systemic infection. For successful establishment of infection, L. monocytogenes must survive and adapt to the low pH environment of the stomach. Gene sequence analysis indicates that lmo0043, an orthologue of arcA, encodes a protein containing conserved motifs and critical active amino acids characteristic of arginine deiminase that mediates an arginine deimination reaction. We attempted to characterize the role of ArcA in acid tolerance in vitro and in mice models. Transcription of arcA was significantly increased in L. monocytogenes culture subjected to acid stress at pH 4.8, as compared with that at pH 7.0. Deletion of arcA impaired growth of L. monocytogenes under mild acidic conditions at pH 5.5, and reduced its survival in synthetic human gastric fluid at pH 2.5 and in the murine stomach. Bacterial load in the spleen of mice intraperitoneally inoculated with an arcA deletion mutant was significantly lower than that of the wild-type strain. These phenotypic changes were recoverable by genetic complementation. Thus, we conclude that L. monocytogenes arcA not only mediates acid tolerance in vitro but also participates in gastric survival and virulence in mice.

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