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.

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 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.

Acid tolerance in Listeria monocytogenes influences invasiveness of enterocyte-like cells and macrophage-like cells

2000 ◽  
Vol 29 (3) ◽  
pp. 137-144 ◽  
Author(s):  
MP Conte ◽  
G Petrone ◽  
AM Di Biase ◽  
MG Ammendolia ◽  
F Superti ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Acid Tolerance

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.

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