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.

scholarly journals Lmo0036, an ornithine and putrescine carbamoyltransferase in Listeria monocytogenes, participates in arginine deiminase and agmatine deiminase pathways and mediates acid tolerance

Microbiology ◽  
2011 ◽  
Vol 157 (11) ◽  
pp. 3150-3161 ◽  
Author(s):  
Jianshun Chen ◽  
Changyong Cheng ◽  
Ye Xia ◽  
Hanxin Zhao ◽  
Chun Fang ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Foodborne Pathogen ◽  
Acid Tolerance ◽  
Arginine Deiminase ◽  
Transcriptional Level ◽  
Acidic Conditions ◽  
Acidic Environments ◽  
Agmatine Deiminase

Listeria monocytogenes is a foodborne pathogen causing listeriosis. Acid is one of the stresses that foodborne pathogens encounter most frequently. The ability to survive and proliferate in acidic environments is a prerequisite for infection. However, there is limited knowledge about the molecular basis of adaptation of L. monocytogenes to acid. Arginine deiminase (ADI) and agmatine deiminase (AgDI) systems are implicated in bacterial tolerance to acidic environments. Homologues of ADI and AgDI systems have been found in L. monocytogenes lineages I and II strains. Sequence analysis indicated that lmo0036 encodes a putative carbamoyltransferase containing conserved motifs and residues important for substrate binding. Lmo0036 acted as an ornithine carbamoyltransferase and putrescine carbamoyltransferase, representing the first example, to our knowledge, that catalyses reversible ornithine and putrescine carbamoyltransfer reactions. Catabolic ornithine and putrescine carbamoyltransfer reactions constitute the second step of ADI and AgDI pathways. However, the equilibrium of in vitro carbamoyltransfer reactions was overwhelmingly towards the anabolic direction, suggesting that catabolic carbamoyltransferase was probably the limiting step of the pathways. lmo0036 was induced at the transcriptional level when L. monocytogenes was subjected to low-pH stress. Its expression product in Escherichia coli exhibited higher catabolic carbamoyltransfer activities under acidic conditions. Consistently, absence of this enzyme impaired the growth of Listeria under mild acidic conditions (pH 4.8) and reduced its survival in synthetic human gastric fluid (pH 2.5), and corresponded to a loss in ammonia production, indicating that Lmo0036 was responsible for acid tolerance at both sublethal and lethal pH levels. Furthermore, Lmo0036 played a possible role in Listeria virulence.

scholarly journals Transcytosis of Listeria monocytogenes across the intestinal barrier upon specific targeting of goblet cell accessible E-cadherin

2011 ◽  
Vol 208 (11) ◽  
pp. 2263-2277 ◽  
Author(s):  
Georgios Nikitas ◽  
Chantal Deschamps ◽  
Olivier Disson ◽  
Théodora Niault ◽  
Pascale Cossart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Intestinal Epithelium ◽  
Foodborne Pathogen ◽  
Intestinal Barrier ◽  
Systemic Infection ◽  
Surface Protein ◽  
Intestinal Lumen ◽  
Specific Host ◽  
Species Specific ◽  
E Cadherin

Listeria monocytogenes (Lm) is a foodborne pathogen that crosses the intestinal barrier upon interaction between its surface protein InlA and its species-specific host receptor E-cadherin (Ecad). Ecad, the key constituent of adherens junctions, is typically situated below tight junctions and therefore considered inaccessible from the intestinal lumen. In this study, we investigated how Lm specifically targets its receptor on intestinal villi and crosses the intestinal epithelium to disseminate systemically. We demonstrate that Ecad is luminally accessible around mucus-expelling goblet cells (GCs), around extruding enterocytes at the tip and lateral sides of villi, and in villus epithelial folds. We show that upon preferential adherence to accessible Ecad on GCs, Lm is internalized, rapidly transcytosed across the intestinal epithelium, and released in the lamina propria by exocytosis from where it disseminates systemically. Together, these results show that Lm exploits intrinsic tissue heterogeneity to access its receptor and reveal transcytosis as a novel and unanticipated pathway that is hijacked by Lm to breach the intestinal epithelium and cause systemic infection.

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.

scholarly journals Contribution of dps to Acid Stress Tolerance and Oxidative Stress Tolerance in Escherichia coli O157:H7

2000 ◽  
Vol 66 (9) ◽  
pp. 3911-3916 ◽  
Author(s):  
Sang Ho Choi ◽  
David J. Baumler ◽  
Charles W. Kaspar
Keyword(s):  
Oxidative Stress ◽  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Stationary Phase ◽  
Stress Tolerance ◽  
Parent Strain ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Gastric Fluid ◽  
Acid Challenge

ABSTRACT An Escherichia coli O157:H7dps::nptI mutant (FRIK 47991) was generated, and its survival was compared to that of the parent in HCl (synthetic gastric fluid, pH 1.8) and hydrogen peroxide (15 mM) challenges. The survival of the mutant in log phase (5-h culture) was significantly impaired (4-log10-CFU/ml reduction) compared to that of the parent strain (ca. 1.0-log10-CFU/ml reduction) after a standard 3-h acid challenge. Early-stationary-phase cells (12-h culture) of the mutant decreased by ca. 4 log10CFU/ml while the parent strain decreased by approximately 2 log10 CFU/ml. No significant differences in the survival of late-stationary-phase cells (24-h culture) between the parent strain and the mutant were observed, although numbers of the parent strain declined less in the initial 1 h of acid challenge. FRIK 47991 was more sensitive to hydrogen peroxide challenge than was the parent strain, although survival improved in stationary phase. Complementation of the mutant with a functional dps gene restored acid and hydrogen peroxide tolerance to levels equal to or greater than those exhibited by the parent strain. These results demonstrate that decreases in survival were from the absence of Dps or a protein regulated by Dps. The results from this study establish that Dps contributes to acid tolerance in E. coli O157:H7 and confirm the importance of Dps in oxidative stress protection.

Comparison of the Stress Response of Listeria monocytogenes Strains with Sprout Colonization

2008 ◽  
Vol 71 (8) ◽  
pp. 1556-1562 ◽  
Author(s):  
LISA GORSKI ◽  
DENISE FLAHERTY ◽  
JESSICA M. DUHÉ
Keyword(s):  
Oxidative Stress ◽  
Statistical Analysis ◽  
Listeria Monocytogenes ◽  
Stress Response ◽  
Foodborne Pathogen ◽  
Acid Stress ◽  
Broccoli Sprouts ◽  
Alfalfa Sprouts ◽  
Different Strains ◽  
And Oxidative Stress

Twenty-nine strains of the foodborne pathogen Listeria monocytogenes were tested for their ability to colonize alfalfa, radish, and broccoli sprouts and their capacity to withstand acid and oxidative stress, two stresses common to the sprouting environment. Wide variation in the ability of different strains to colonize alfalfa sprouts were confirmed, but the variations among radish and broccoli sprouts were not as large. With a few exceptions, strains that were poor colonizers of alfalfa tended to be among the poorer colonizers of radish and broccoli and vice versa. The strains also were variable in their resistance to both acid and oxidative stress. Statistical analysis revealed no correlation between acid stress and sprout colonization, but there was a positive correlation between resistance to oxidative stress and colonization of all three sprout types. Although the response to oxidative stress is important for L. monocytogenes virulence, it also may be important for life outside of a host.

scholarly journals Mutant and Recombinant Phages Selected from In Vitro Coevolution Conditions Overcome Phage-Resistant Listeria monocytogenes

2020 ◽  
Vol 86 (22) ◽  
Author(s):  
Tracey Lee Peters ◽  
Yaxiong Song ◽  
Daniel W. Bryan ◽  
Lauren K. Hudson ◽  
Thomas G. Denes
Keyword(s):  
Listeria Monocytogenes ◽  
Host Range ◽  
Foodborne Pathogen ◽  
Resistant Bacteria ◽  
Phage Resistance ◽  
Short Term ◽  
Content Type ◽  
Life Threatening ◽  
The Impact

ABSTRACT Bacteriophages (phages) are currently available for use by the food industry to control the foodborne pathogen Listeria monocytogenes. Although phage biocontrols are effective under specific conditions, their use can select for phage-resistant bacteria that repopulate phage-treated environments. Here, we performed short-term coevolution experiments to investigate the impact of single phages and a two-phage cocktail on the regrowth of phage-resistant L. monocytogenes and the adaptation of the phages to overcome this resistance. We used whole-genome sequencing to identify mutations in the target host that confer phage resistance and in the phages that alter host range. We found that infections with Listeria phages LP-048, LP-125, or a combination of both select for different populations of phage-resistant L. monocytogenes bacteria with different regrowth times. Phages isolated from the end of the coevolution experiments were found to have gained the ability to infect phage-resistant mutants of L. monocytogenes and L. monocytogenes strains previously found to be broadly resistant to phage infection. Phages isolated from coinfected cultures were identified as recombinants of LP-048 and LP-125. Interestingly, recombination events occurred twice independently in a locus encoding two proteins putatively involved in DNA binding. We show that short-term coevolution of phages and their hosts can be utilized to obtain mutant and recombinant phages with adapted host ranges. These laboratory-evolved phages may be useful for limiting the emergence of phage resistance and for targeting strains that show general resistance to wild-type (WT) phages. IMPORTANCE Listeria monocytogenes is a life-threatening bacterial foodborne pathogen that can persist in food processing facilities for years. Phages can be used to control L. monocytogenes in food production, but phage-resistant bacterial subpopulations can regrow in phage-treated environments. Coevolution experiments were conducted on a Listeria phage-host system to provide insight into the genetic variation that emerges in both the phage and bacterial host under reciprocal selective pressure. As expected, mutations were identified in both phage and host, but additionally, recombination events were shown to have repeatedly occurred between closely related phages that coinfected L. monocytogenes. This study demonstrates that in vitro evolution of phages can be utilized to expand the host range and improve the long-term efficacy of phage-based control of L. monocytogenes. This approach may also be applied to other phage-host systems for applications in biocontrol, detection, and phage therapy.

scholarly journals Streptococcus thermophiles DMST-H2 Promotes Recovery in Mice with Antibiotic-Associated Diarrhea

2020 ◽  
Vol 8 (11) ◽  
pp. 1650
Author(s):  
Jin-Shuang Hu ◽  
Yan-Yan Huang ◽  
Jia-Hua Kuang ◽  
Jia-Jia Yu ◽  
Qin-Yu Zhou ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
Intestinal Flora ◽  
Intestinal Barrier ◽  
Genomic Analysis ◽  
Acid Tolerance ◽  
Interferon Γ ◽  
Common Side Effect ◽  
Gastrointestinal Conditions ◽  
Antibiotic Associated Diarrhea

Antibiotic-associated diarrhea (AAD) is the most common side effect of antibiotics and is routinely treated with probiotics in clinical. Streptococcus thermophiles, extensively utilized for producing dairy foods, has recently been regarded as a new promising probiotic candidate. In this study, the efficacy of Streptococcus thermophiles DMST-H2 (DMST-H2) for AAD treatment in mice was investigated. DMST-H2 was isolated from Chinese traditional yogurt, proved to be non-toxic, and presented tolerance against simulated gastrointestinal conditions in vitro. Additionally, genomic analysis revealed that it possessed genes related to acid tolerance, bile salt tolerance, adhesion, oxidative stress and bacteriocin production. The animal experiment results showed that both DMST-H2 treatment and natural recovery could reduce fecal water content. Compared with spontaneous recovery, DMST-H2 accelerated the recovery of the enlarged caecum and intestinal barrier injury from AAD, and further decreased endotoxin (ET), D-lactate (D-LA) and diamine oxidase (DAO) content in serum. Moreover, pro-inflammatory cytokines (TNF-α) were reduced, while interferon-γ (IFN-γ) and anti-inflammatory cytokines (IL-10) increased after treating with DMST-H2. Furthermore, DMST-H2 better restored the structure of intestinal flora. At the phylum level, Firmicutes increased and Proteobacteria decreased. These findings indicate that DMST-H2 could promote recovery in mice with antibiotic-associated diarrhea.

scholarly journals β-Phenylethylamine as a Natural Food Additive Shows Antimicrobial Activity against Listeria monocytogenes on Ready-to-Eat Foods

Foods ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 1363 ◽  
Author(s):  
Francis Muchaamba ◽  
Roger Stephan ◽  
Taurai Tasara
Keyword(s):  
Listeria Monocytogenes ◽  
Foodborne Pathogen ◽  
Bactericidal Effect ◽  
Model Systems ◽  
Natural Food ◽  
Smoked Salmon ◽  
Growth Inhibitory ◽  
Good Manufacturing Practices ◽  
Agar Media

Listeria monocytogenes is an important foodborne pathogen and a major cause of death associated with bacterial foodborne infections. Control of L. monocytogenes on most ready-to-eat (RTE) foods remains a challenge. The potential use of β-phenylethylamine (PEA) as an organic antimicrobial against L. monocytogenes was evaluated in an effort to develop a new intervention for its control. Using a collection of 62 clinical and food-related isolates we determined the minimum inhibitory concentration (MIC) of PEA against L. monocytogenes in different broth and agar media. Bologna type sausage (lyoner) and smoked salmon were used as food model systems to validate the in vitro findings. PEA had a growth inhibitory and bactericidal effect against L. monocytogenes both in in vitro experiments as well as on lyoner and smoked salmon. The MIC’s ranged from 8 to 12.5 mg/mL. Furthermore, PEA also inhibited L. monocytogenes biofilm formation. Based on good manufacturing practices as a prerequisite, the application of PEA to RTE products might be an additional hurdle to limit L. monocytogenes growth thereby increasing food safety.

scholarly journals Streptococcus pyogenes Arginine and Citrulline Catabolism Promotes Infection and Modulates Innate Immunity

2013 ◽  
Vol 82 (1) ◽  
pp. 233-242 ◽  
Author(s):  
Zachary T. Cusumano ◽  
Michael E. Watson ◽  
Michael G. Caparon
Keyword(s):  
Nitric Oxide ◽  
Innate Immunity ◽  
Streptococcus Pyogenes ◽  
Murine Model ◽  
Acid Stress ◽  
Arginine Deiminase ◽  
Content Type ◽  
Cutaneous Tissue

ABSTRACTA bacterium's ability to acquire nutrients from its host during infection is an essential component of pathogenesis. For the Gram-positive pathogenStreptococcus pyogenes, catabolism of the amino acid arginine via the arginine deiminase (ADI) pathway supplements energy production and provides protection against acid stressin vitro. Its expression is enhanced in murine models of infection, suggesting an important rolein vivo. To gain insight into the function of the ADI pathway in pathogenesis, the virulence of mutants defective in each of its enzymes was examined. Mutants unable to use arginine (ΔArcA) or citrulline (ΔArcB) were attenuated for carriage in a murine model of asymptomatic mucosal colonization. However, in a murine model of inflammatory infection of cutaneous tissue, the ΔArcA mutant was attenuated but the ΔArcB mutant was hyperattenuated, revealing an unexpected tissue-specific role for citrulline metabolism in pathogenesis. When mice defective for the arginine-dependent production of nitric oxide (iNOS−/−) were infected with the ΔArcA mutant, cutaneous virulence was rescued, demonstrating that the ability ofS. pyogenesto utilize arginine was dispensable in the absence of nitric oxide-mediated innate immunity. This work demonstrates the importance of arginine and citrulline catabolism and suggests a novel mechanism of virulence by whichS. pyogenesuses its metabolism to modulate innate immunity through depletion of an essential host nutrient.

Survival of Listeria monocytogenes during Production and Ripening of Traditional Turkish Savak Tulum Cheese and in Synthetic Gastric Fluid

2013 ◽  
Vol 76 (10) ◽  
pp. 1801-1805 ◽  
Author(s):  
ABDULLAH DIKICI ◽  
MEHMET CALICIOGLU
Keyword(s):  
Listeria Monocytogenes ◽  
Acid Tolerance ◽  
Gastric Fluid ◽  
Chemical Analyses ◽  
Tolerance Mechanisms ◽  
Processing Stage ◽  
Ripening Period ◽  
Production Stage ◽  
Traditional Cheese ◽  
Tulum Cheese

This study investigated the survival and acid tolerance of Listeria monocytogenes during the 2-day processing stage and 90-day ripening of Savak tulum cheese, a traditional cheese in Turkey. Experimental Savak tulum cheese was produced from raw sheep's milk that was inoculated with a L. monocytogenes mixture consisting of five strains (average 7.0 log CFU/ml) and was ripened at 6°C for 90 days. Microbiological and chemical analyses and acid exposure experiments in synthetic gastric fluid (SGF) (pH 1.5 to 2.5) were carried out on days 1 and 2 during processing and on days 0, 15, 30, 45, 60, and 90 during ripening. The numbers of L. monocytogenes did not decrease during processing, but a total of 4.1 log CFU/g reduction was observed during ripening. Throughout the ripening period, L. monocytogenes cells survived direct 90-min exposures of the cheese samples to SGF. These results suggest that, although the pathogen numbers decreased in Savak tulum cheese ripened at 6°C for 90 days, a sublethal environment may have occurred in the cheese during the production stage, activating the acid-tolerance mechanisms of the pathogen and allowing L. monocytogenes to maintain its viability in the SGF for 90 min.

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