scholarly journals Anaerobic growth of Listeria monocytogenes on rhamnose is stimulated by Vitamin B12 and bacterial microcompartment dependent 1,2-propanediol utilization

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Siming Li ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Vitamin B12 ◽  
Anaerobic Growth ◽  
Physiological Effects ◽  
Human Intestine ◽  
Human Pathogens ◽  
Bacterial Microcompartment ◽  
Food Borne ◽  
Transmission Electron ◽  
Shell Proteins

AbstractThe food-borne pathogen Listeria monocytogenes is able to form proteinaceous organelles called bacterial microcompartments (BMCs) that optimize the utilization of substrates, such as 1,2-propanediol, and confer an anaerobic growth advantage. Rhamnose is a deoxyhexose sugar abundant in a range of environments including the human intestine, and can be degraded in anaerobic conditions into 1,2-propanediol, next to acetate and lactate. Rhamnose-derived 1,2-propanediol has been found to link with BMCs in a limited number of commensal human colonic species and some human pathogens such as Salmonella enterica, but the involvement of BMCs in rhamnose metabolism and potential physiological effects on L. monocytogenes are still unknown. In this study, we firstly test the effect of rhamnose uptake and utilization on anaerobic growth of L. monocytogenes EGDe without and with added vitamin B12, followed by metabolic analysis. We unveil that the vitamin B12-dependent activation of pdu stimulates metabolism and anaerobic growth of L. monocytogenes EGDe on rhamnose via 1,2-propanediol degradation into 1-propanol and propionate. Transmission electron microscopy of pdu-induced cells shows that BMCs are formed and additional proteomics experiments confirm expression of pdu BMC shell proteins and enzymes. Finally, we discuss physiological effects and energy efficiency of L. monocytogenes pdu BMC-driven anaerobic rhamnose metabolism and impact on competitive fitness in environments such as the human intestine.

scholarly journals Impact of vitamin B12 on rhamnose metabolism, stress defense and in-vitro virulence of Listeria monocytogenes

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Lucas M. Wijnands ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Vitamin B12 ◽  
Growth Stimulation ◽  
Anaerobic Growth ◽  
Salmonella Spp ◽  
Defense Proteins ◽  
Bacterial Microcompartment ◽  
The Impact ◽  
Stress Defense

Listeria monocytogenes is a facultative anaerobe which can cause a severe food-borne infection known as listeriosis. Rhamnose is a deoxyhexose sugar abundant in a range of environments, including the human intestine, and can be degraded by L. monocytogenes in aerobic and anaerobic conditions into lactate, acetate and 1,2-propanediol. Our previous study showed that addition of vitamin B12 stimulates anaerobic growth of L. monocytogenes on rhamnose due to the activation of bacterial microcompartment (BMC)-dependent 1,2-propanediol utilization with concomitant production of propionate and propanol. Notably, anaerobic propanediol metabolism has been linked to virulence of enteric pathogens including Salmonella spp. and L. monocytogenes. In this study we investigate the impact of B12 on aerobic and anerobic growth of L. monocytogenes on rhamnose, and observed growth stimulation and pdu BMC activation only in anaerobically grown cells with B12 added to the medium. Comparative Caco-2 virulence assays, showed that these pdu BMC induced cells have significantly higher translocation efficiency compared to aerobically grown cells (without and with added B12) and non-induced anaerobically grown cells, while adhesion and invasion capacity is similar for all cells. Comparative proteomics analysis showed specific and overlapping responses linked to metabolic shifts, activation of stress defense proteins and virulence factors, with RNA polymerase sigma factor SigL; teichoic acids export ATP-binding protein, TagH; DNA repair and protection proteins RadA and DPS; and glutathione synthase GshAB previously linked to activation of virulence response in L. monocytogenes, uniquely upregulated in anaerobically rhamnose grown pdu BMC induced cells. Our results shed new light into B12 impact on L. monocytogenes competitive fitness and virulence.

scholarly journals Impact of bacterial microcompartment-dependent ethanolamine and propanediol metabolism on Listeria monocytogenes interactions with Caco-2 cells

2021 ◽  
Author(s):  
Zhe Zeng ◽  
Lucas M. Wijnands ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Human Cells ◽  
Cellular Systems ◽  
Anaerobic Growth ◽  
Commensal Microbiota ◽  
Bacterial Microcompartment ◽  
Transmission Electron ◽  
Well Assay ◽  
Phospholipid Degradation

AbstractBacterial microcompartment (BMC) dependent ethanolamine (eut) and propanediol utilization (pdu) has recently been shown to stimulate anaerobic growth of Listeria monocytogenes. This metabolic repertoire conceivably contributes to the competitive fitness of L. monocytogenes in the human gastrointestinal (GI) tract, where these compounds become available following phospholipid degradation and mucus-derived rhamnose metabolism by commensal microbiota. Previous transcriptomics and mutant studies of eut and pdu L. monocytogenes suggested a possible role of eut and pdu BMC metabolism in transmission in foods and pathogenicity, but data on a potential role of L. monocytogenes interaction with human cells is currently absent. First, we ask which cellular systems are expressed in the activation of eut and pdu BMC metabolism and the extent to which these systems are conserved between the states. We find common and unique systems related to metabolic shifts, stress and virulence factors. Next, we hypothesize that these common and unique activated cellular systems contribute to a role in the interaction of L. monocytogenes interaction with human cells. We present evidence that metabolically primed L. monocytogenes with active eut and pdu BMCs, as confirmed by metabolic analysis, transmission electron microscopy and proteomics, show significantly enhanced translocation efficacy compared to non-induced cells in a trans-well assay using Caco-2 cells, while adhesion and invasion capacity was similar. Taken together, our results provide insights into the possible key cellular players that drive translocation efficacy upon eut and pdu BMC activation.

scholarly journals Bacterial Microcompartment-Dependent 1,2-Propanediol Utilization of Propionibacterium freudenreichii

2021 ◽  
Vol 12 ◽  
Author(s):  
Alexander Dank ◽  
Zhe Zeng ◽  
Sjef Boeren ◽  
Richard A. Notebaart ◽  
Eddy J. Smid ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
Probiotic Bacterium ◽  
Vitamin B ◽  
Propionibacterium Freudenreichii ◽  
Human Gut ◽  
Bacterial Microcompartment ◽  
Gut Colonization ◽  
Transmission Electron ◽  
Shell Proteins ◽  
Repair Mechanisms

Bacterial microcompartments (BMCs) are proteinaceous prokaryotic organelles that enable the utilization of substrates such as 1,2-propanediol and ethanolamine. BMCs are mostly linked to the survival of particular pathogenic bacteria by providing a growth advantage through utilization of 1,2-propanediol and ethanolamine which are abundantly present in the human gut. Although a 1,2-propanediol utilization cluster was found in the probiotic bacterium Propionibacterium freudenreichii, BMC-mediated metabolism of 1,2-propanediol has not been demonstrated experimentally in P. freudenreichii. In this study we show that P. freudenreichii DSM 20271 metabolizes 1,2-propanediol in anaerobic conditions to propionate and 1-propanol. Furthermore, 1,2-propanediol induced the formation of BMCs, which were visualized by transmission electron microscopy and resembled BMCs found in other bacteria. Proteomic analysis of 1,2-propanediol grown cells compared to L-lactate grown cells showed significant upregulation of proteins involved in propanediol-utilization (pdu-cluster), DNA repair mechanisms and BMC shell proteins while proteins involved in oxidative phosphorylation were down-regulated. 1,2-Propanediol utilizing cells actively produced vitamin B12 (cobalamin) in similar amounts as cells growing on L-lactate. The ability to metabolize 1,2-propanediol may have implications for human gut colonization and modulation, and can potentially aid in delivering propionate and vitamin B12in situ.

scholarly journals Anaerobic Growth of Listeria monocytogenes on Rhamnose Is Stimulated by Vitamin B 12 and Bacterial Microcompartment-Dependent 1,2-Propanediol Utilization

mSphere ◽  
2021 ◽  
Author(s):  
Zhe Zeng ◽  
Siming Li ◽  
Sjef Boeren ◽  
Eddy J. Smid ◽  
Richard A. Notebaart ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Molecular Mechanisms ◽  
Foodborne Pathogen ◽  
Anaerobic Growth ◽  
Severe Illness ◽  
Vitamin B ◽  
Anaerobic Conditions ◽  
Content Type ◽  
Bacterial Microcompartment ◽  
Vitamin B 12

Listeria monocytogenes is a foodborne pathogen causing severe illness and, as such, it is crucial to understand the molecular mechanisms contributing to its survival strategy and pathogenicity. Rhamnose is a deoxyhexose sugar abundant in a range of environments, including the human intestine, and can be degraded in anaerobic conditions into 1,2-propanediol.

scholarly journals Biocontrol of the Food-Borne Pathogens Listeria monocytogenes and Salmonella enterica Serovar Poona on Fresh-Cut Apples with Naturally Occurring Bacterial and Yeast Antagonists

2006 ◽  
Vol 72 (2) ◽  
pp. 1135-1140 ◽  
Author(s):  
Britta Leverentz ◽  
William S. Conway ◽  
Wojciech Janisiewicz ◽  
Maribel Abadias ◽  
Cletus P. Kurtzman ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Salmonella Enterica ◽  
Human Pathogens ◽  
Food Borne Pathogens ◽  
Naturally Occurring ◽  
Food Borne ◽  
Different Temperatures ◽  
Apple Tissue ◽  
Higher Temperature ◽  
Fresh Cut

ABSTRACT Fresh-cut apples contaminated with either Listeria monocytogenes or Salmonella enterica serovar Poona, using strains implicated in outbreaks, were treated with one of 17 antagonists originally selected for their ability to inhibit fungal postharvest decay on fruit. While most of the antagonists increased the growth of the food-borne pathogens, four of them, including Gluconobacter asaii (T1-D1), a Candida sp. (T4-E4), Discosphaerina fagi (ST1-C9), and Metschnikowia pulcherrima (T1-E2), proved effective in preventing the growth or survival of food-borne human pathogens on fresh-cut apple tissue. The contaminated apple tissue plugs were stored for up to 7 days at two different temperatures. The four antagonists survived or grew on the apple tissue at 10 or 25°C. These four antagonists reduced the Listeria monocytogenes populations and except for the Candida sp. (T4-E4), also reduced the S. enterica serovar Poona populations. The reduction was higher at 25°C than at 10°C, and the growth of the antagonists, as well as pathogens, increased at the higher temperature.

scholarly journals Transmission Electron – Microscopic Study of Listeria monocytogenes isolated from raw milk and soft cheese in Baghdad Province

2012 ◽  
Vol 6 (1) ◽  
pp. 5-9
Author(s):  
Ali Hassan Ahmed AL-Shamary
Keyword(s):  
Listeria Monocytogenes ◽  
Raw Milk ◽  
Food Microbiology ◽  
Secretion Systems ◽  
Electron Microscopic ◽  
Transmission Electron Microscopic Study ◽  
Food Borne ◽  
Transmission Electron ◽  
Soft Cheese ◽  
Type 3

The study was designed to confirm and understand the internal and external features of Listeria monocytogenes by Transmission electron microscopy (TEM). A study was conducted at the College of Veterinary Medicine/ University of Baghdad and College of Medicine/AL-Nahrain University during April-May 2008. Standard listerial broth was prepared in food Lab. at Baghdad Vet. Coll. from locally isolated strains of Listeria monocytogenes (from raw milk and soft cheese in Baghdad province) according to standard protocols of food microbiology, transmitted in refrigerated containers to TEM-section at AL-Nahrain Coll. of Med. for processing and photographing the different forms and components of isolates. The results revealed polyforms and components of Listeria monocytogenes that important for diagnosis and therapy such as V-shape phenomenon in listerial cells, periplasmic space, DNA, plasmids, pathogenicity islands, type 3 secretion systems, and sausage chain in listerial cells (Incomplete-dissociation). In conclusion TEM technology is an important tool for diagnosing of zoonotic food borne diseases between animals and man.

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.

scholarly journals Effect of Gaseous Ozone on Listeria monocytogenes Planktonic Cells and Biofilm: An In Vitro Study

Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1484
Author(s):  
Felice Panebianco ◽  
Selene Rubiola ◽  
Francesco Chiesa ◽  
Tiziana Civera ◽  
Pierluigi Aldo Di Ciccio
Keyword(s):  
Listeria Monocytogenes ◽  
In Vitro Study ◽  
Planktonic Cells ◽  
Free Living ◽  
Biofilm Inhibition ◽  
Additional Tool ◽  
Complete Inactivation ◽  
Gaseous Ozone ◽  
Food Borne

Among food-borne pathogens, Listeria monocytogenes continues to pose concerns to food business operators due to its capacity to form biofilm in processing environments. Ozone may be an eco-friendly technology to control microbial contaminations, but data concerning its effect on Listeria monocytogenes biofilm are still limited. In this study, the effect of gaseous ozone at 50 ppm on planktonic cells and biofilm of reference and food-related Listeria monocytogenes strains was evaluated. Ozone caused a reduction in microbial loads of 3.7 ± 0.4 and 3.9 ± 0.4 Log10 CFU/mL after 10 and 30 min, respectively. A complete inactivation of planktonic cells after 6 h of treatment was observed. Biofilm inhibition and eradication treatments (50 ppm, 6 h) resulted in a significant decrease of the biofilm biomass for 59% of the strains tested, whilst a slight dampening of live cell loads in the biofilm state was observed. In conclusion, gaseous ozone is not sufficient to completely counteract Listeria monocytogenes biofilm, but it may be useful as an additional tool to contrast Listeria monocytogenes free-living cells and to improve the existing sanitization procedures in food processing environments.

scholarly journals Gbu Glycine Betaine Porter and Carnitine Uptake in Osmotically Stressed Listeria monocytogenes Cells

2002 ◽  
Vol 68 (11) ◽  
pp. 5647-5655 ◽  
Author(s):  
Mary Lou Mendum ◽  
Linda Tombras Smith
Keyword(s):  
Listeria Monocytogenes ◽  
Transport System ◽  
Glycine Betaine ◽  
Compatible Solutes ◽  
High Salt ◽  
Uptake System ◽  
Food Borne ◽  
Protein Mutants ◽  
Carnitine Uptake ◽  
Do So

ABSTRACT The food-borne pathogen Listeria monocytogenes grows actively under high-salt conditions by accumulating compatible solutes such as glycine betaine and carnitine from the medium. We report here that the dominant transport system for glycine betaine uptake, the Gbu porter, may act as a secondary uptake system for carnitine, with a Km of 4 mM for carnitine uptake and measurable uptake at carnitine concentrations as low as 10 μM. This porter has a Km for glycine betaine uptake of about 6 μM. The dedicated carnitine porter, OpuC, has a Km for carnitine uptake of 1 to 3 μM and a V max of approximately 15 nmol/min/mg of protein. Mutants lacking either opuC or gbu were used to study the effects of four carnitine analogs on growth and uptake of osmolytes. In strain DP-L1044, which had OpuC and the two glycine betaine porters Gbu and BetL, triethylglycine was most effective in inhibiting growth in the presence of glycine betaine, but trigonelline was best at inhibiting growth in the presence of carnitine. Carnitine uptake through OpuC was inhibited by γ-butyrobetaine. Dimethylglycine inhibited both glycine betaine and carnitine uptake through the Gbu porter. Carnitine uptake through the Gbu porter was inhibited by triethylglycine. Glycine betaine uptake through the BetL porter was strongly inhibited by trigonelline and triethylglycine. These results suggest that it is possible to reduce the growth of L. monocytogenes under osmotically stressful conditions by inhibiting glycine betaine and carnitine uptake but that to do so, multiple uptake systems must be affected.

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