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.

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 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 Impedance Sensing Platform for Detection of the Food Pathogen Listeria monocytogenes

Electronics ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 347 ◽  
Author(s):  
Maria Chiriacò ◽  
Ilaria Parlangeli ◽  
Fausto Sirsi ◽  
Palmiro Poltronieri ◽  
Elisabetta Primiceri
Keyword(s):  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Foodborne Pathogen ◽  
High Sensitivity ◽  
Primary Objective ◽  
Field Application ◽  
Impedance Sensing ◽  
Sensing Platform

A great improvement in food safety and quality controls worldwide has been achieved through the development of biosensing platforms. Foodborne pathogens continue to cause serious outbreaks, due to the ingestion of contaminated food. The development of new, sensitive, portable, high-throughput, and automated platforms is a primary objective to allow detection of pathogens and their toxins in foods. Listeria monocytogenes is one common foodborne pathogen. Major outbreaks of listeriosis have been caused by a variety of foods, including milk, soft cheeses, meat, fermented sausages, poultry, seafood and vegetable products. Due to its high sensitivity and easy setup, electrochemical impedance spectroscopy (EIS) has been extensively applied for biosensor fabrication and in particular in the field of microbiology as a mean to detect and quantify foodborne bacteria. Here we describe a miniaturized, portable EIS platform consisting of a microfluidic device with EIS sensors for the detection of L. monocytogenes in milk samples, connected to a portable impedance analyzer for on-field application in clinical and food diagnostics, but also for biosecurity purposes. To achieve this goal microelectrodes were functionalized with antibodies specific for L. monocytogenes. The binding and detection of L. monocytogenes was achieved in the range 2.2 × 103 cfu/mL to 1 × 102 with a Limit of Detection (LoD) of 5.5 cfu/mL.

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 The Inhibitory Effect of Plant Extracts on Growth of the Foodborne Pathogen, Listeria monocytogenes

Antibiotics ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 319 ◽  
Author(s):  
Marina Ceruso ◽  
Jason A. Clement ◽  
Matthew J. Todd ◽  
Fangyuan Zhang ◽  
Zuyi Huang ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Plant Extracts ◽  
Foodborne Pathogens ◽  
Food Industry ◽  
Foodborne Pathogen ◽  
The United States ◽  
Inhibitory Effect ◽  
Microscopic Level ◽  
Natural Antimicrobials ◽  
Minimal Inhibitory Concentrations

Listeria monocytogenes is a foodborne pathogen responsible for about 1600 illnesses each year in the United States (US) and about 2500 confirmed invasive human cases in European Union (EU) countries. Several technologies and antimicrobials are applied to control the presence of L. monocytogenes in food. Among these, the use of natural antimicrobials is preferred by consumers. This is due to their ability to inhibit the growth of foodborne pathogens but not prompt negative safety concerns. Among natural antimicrobials, plant extracts are used to inactivate L. monocytogenes. However, there is a large amount of these types of extracts, and their active compounds remain unexplored. The aim of this study was to evaluate the antibacterial activity against L. monocytogenes of about 800 plant extracts derived from plants native to different countries worldwide. The minimal inhibitory concentrations (MICs) were determined, and scanning electron microscopy (SEM) was used to verify how the plant extracts affected L. monocytogenes at the microscopic level. Results showed that 12 of the plant extracts had inhibitory activity against L. monocytogenes. Future applications of this study could include the use of these plant extracts as new preservatives to reduce the risk of growth of pathogens and contamination in the food industry from L. monocytogenes.

scholarly journals Antimicrobial Activity of Nisin and Lysozyme on Foodborne Pathogens Listeria Monocytogenes, Staphylococcus Aureus, Salmonella Typhimurium, and Escherichia Coli at Different pH

2018 ◽  
Author(s):  
Hamdollah Moshtaghi ◽  
Azadeh Rashidimehr ◽  
Behzad Shareghi
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Inhibitory Concentration ◽  
Low Ph ◽  
Bacteria Growth ◽  
Microdilution Method ◽  
Natural Preservatives

Background: To prevent or inhibit the growth of pathogenic microorganisms and food spoilage factors, many studies have been done by using natural preservatives. The aim of study was to investigate the effect of different concentrations of lysozyme and Nisin on the growth rate and also to determine the minimum inhibitory concentration (MIC) and minimum bactericidal cocentratiin (MBC) of these combinations on the bacteria of Escherichia coli, Staphylococcus aureus, Salmonella typhimorium and Listeria monocytogenes. Methods: In this study, various concentrations of lysozyme and Nisin were set in form of alone concentration and in combination concentrations (0, 19.53, 39.06, 78.13, 156.25, 312.5, 625, 1250, 2500, 5000) in vitro conditions and 6 pH 5.5, 6, 6.5, 7, 7.5, and 8. Microdilution method at 24°C was done and the combined effect on bacteria growth was read by using ELISA reader. Results: The results showed that lysozyme was less effective on Escherichia coli and Nisin was less effective on Listeria monocytogenes. Moreover, combining lysozyme and Nisin at low pH decreased the MIC. Conclusions: The results of the study showed that the effect of combining lysozyme and Nisin on Staphylococcus aureus is above all other bacteria and at low pH reduces the MIC.

scholarly journals Biocide-Tolerant Listeria monocytogenes Isolates from German Food Production Plants Do Not Show Cross-Resistance to Clinically Relevant Antibiotics

2019 ◽  
Vol 85 (20) ◽  
Author(s):  
A. Roedel ◽  
R. Dieckmann ◽  
H. Brendebach ◽  
J. A. Hammerl ◽  
S. Kleta ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Listeria Monocytogenes ◽  
Foodborne Pathogens ◽  
Food Production ◽  
Cross Resistance ◽  
High Genetic Diversity ◽  
Content Type ◽  
Processing Plants ◽  
German Food

ABSTRACT Contamination of food during processing is recognized as a main transmission route of Listeria monocytogenes. To prevent microbial contamination, biocides are widely applied as disinfectants in food processing plants. However, there are concerns about the development of antimicrobial resistance in foodborne pathogens due to widespread biocide usage. In our study, 93 L. monocytogenes isolates from German food production facilities were (i) tested for biocide and antibiotic susceptibility using broth microdilution assays, (ii) analyzed for links between reduced biocide susceptibility and antibiotic resistance, and (iii) characterized by whole-genome sequencing, including the detection of genes coding for biocide tolerance, antibiotic resistance, and other virulence factors. Fifteen L. monocytogenes isolates were tolerant to benzalkonium chloride (BAC), and genes conferring BAC tolerance were found in 13 of them. Antibiotic resistance was not associated with biocide tolerance. BAC-tolerant isolates were assigned to 6 multilocus sequence type (MLST) clonal complexes, and most of them harbored internalin A pseudogenes with premature stop codons or deletions (n = 9). Our study demonstrated a high genetic diversity among the investigated isolates including genotypes that are frequently involved in human infections. Although in vitro adaptation studies to biocides have raised concerns about increasing cross-resistance to antibiotics, our results do not provide evidence for this phenomenon in field isolates. IMPORTANCE Foodborne pathogens such as L. monocytogenes can persist in food production environments for a long time, causing perennial outbreaks. Hence, bacterial pathogens are able to survive cleaning and disinfection procedures. Accordingly, they may be repeatedly exposed to sublethal concentrations of disinfectants, which might result in bacterial adaptation to these biocides. Furthermore, antibiotic coresistance and cross-resistance are known to evolve under biocide selection pressure in vitro. Hence, antimicrobial tolerance seems to play a crucial role in the resilience and persistence of foodborne pathogens in the food chain and might reduce therapeutic options in infectious diseases.

scholarly journals Commensal microbes provide first line defense against Listeria monocytogenes infection

2017 ◽  
Vol 214 (7) ◽  
pp. 1973-1989 ◽  
Author(s):  
Simone Becattini ◽  
Eric R. Littmann ◽  
Rebecca A. Carter ◽  
Sohn G. Kim ◽  
Sejal M. Morjaria ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Intestinal Microbiota ◽  
Systemic Disease ◽  
Foodborne Pathogen ◽  
Antibiotic Administration ◽  
Defensive Role ◽  
Listeria Monocytogenes Infection ◽  
Dose Oral

Listeria monocytogenes is a foodborne pathogen that causes septicemia, meningitis and chorioamnionitis and is associated with high mortality. Immunocompetent humans and animals, however, can tolerate high doses of L. monocytogenes without developing systemic disease. The intestinal microbiota provides colonization resistance against many orally acquired pathogens, and antibiotic-mediated depletion of the microbiota reduces host resistance to infection. Here we show that a diverse microbiota markedly reduces Listeria monocytogenes colonization of the gut lumen and prevents systemic dissemination. Antibiotic administration to mice before low dose oral inoculation increases L. monocytogenes growth in the intestine. In immunodeficient or chemotherapy-treated mice, the intestinal microbiota provides nonredundant defense against lethal, disseminated infection. We have assembled a consortium of commensal bacteria belonging to the Clostridiales order, which exerts in vitro antilisterial activity and confers in vivo resistance upon transfer into germ free mice. Thus, we demonstrate a defensive role of the gut microbiota against Listeria monocytogenes infection and identify intestinal commensal species that, by enhancing resistance against this pathogen, represent potential probiotics.

Use of Epidemiologic and Food Survey Data To Estimate a Purposefully Conservative Dose-Response Relationship for Listeria monocytogenes Levels and Incidence of Listeriosis†

1997 ◽  
Vol 60 (8) ◽  
pp. 918-922 ◽  
Author(s):  
ROBERT L. BUCHANAN ◽  
WILLIAM G. DAMERT ◽  
RICHARD C. WHITING ◽  
MICHAEL van SCHOTHORST
Keyword(s):  
Listeria Monocytogenes ◽  
Survey Data ◽  
Dose Response ◽  
Foodborne Pathogens ◽  
Foodborne Pathogen ◽  
Significant Risk ◽  
Biological Agent ◽  
Quantitative Microbial Risk Assessment ◽  
Response Relationship ◽  
Dose Response Relationship

The development of effective quantitative microbial risk-assessment models for foodborne pathogens depends on the availability of data on the consumers' exposure to a biological agent and the dose-response relationship that relates levels of the biological agent ingested with frequency of infection or disease. Information on the latter has historically been acquired from human volunteer feeding studies. However, such studies are not feasible for pathogens that either have a significant risk of being life threatening or for which morbidity is primarily associated with high-risk populations (i.e., immunocompromised persons). For these pathogens, it is proposed that purposefully conservative dose-response relationships can be estimated on the basis of combining available epidemiologic data with food-survey data for a ready-to-eat product. As an example, data on the incidence of listeriosis in Germany were combined with data on the levels of Listeria monocytogenes in smoked fish to generate a dose-response curve for this foodborne pathogen.

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