0000000000496106
AUTHOR
Pascal Piveteau
Biotic and abiotic soil properties influence survival of Listeria monocytogenes in soil
International audience; Listeria monocytogenes is a food-borne pathogen responsible for the potentially fatal disease listeriosis and terrestrial ecosystems have been hypothesized to be its natural reservoir. Therefore, identifying the key edaphic factors that influence its survival in soil is critical. We measured the survival of L. monocytogenes in a set of 100 soil samples belonging to the French Soil Quality Monitoring Network. This soil collection is meant to be representative of the pedology and land use of the whole French territory. The population of L. monocytogenes in inoculated microcosms was enumerated by plate count after 7, 14 and 84 days of incubation. Analysis of survival pr…
Biofilm formation of listeria monocytogenes strains under food processing environments and pan-genome-wide association study
International audience; Concerns about food contamination by Listeria monocytogenes are on the rise with increasing consumption of ready-to-eat foods. Biofilm production of L. monocytogenes is presumed to be one of the ways that confer its increased resistance and persistence in the food chain. In this study, a collection of isolates from foods and food processing environments (FPEs) representing persistent, prevalent, and rarely detected genotypes was evaluated for biofilm forming capacities including adhesion and sessile biomass production under diverse environmental conditions. The quantity of sessile biomass varied according to growth conditions, lineage, serotype as well as genotype bu…
Truncated internalin A and asymptomatic Listeria monocytogenes carriage: in vivo investigation by allelic exchange
ABSTRACT Allelic exchange of the region coding for the C terminus of InlA between one epidemic (with an 80-kDa InlA) and one asymptomatic (with a 47-kDa InlA) carriage Listeria monocytogenes strain confirmed the need for this region for internalin entry in vitro. Interestingly, restoration of internalin A functionality did not result in full virulence in chicken embryo assays.
Investigation of root-associated biofilms of the human pathogenic bacterium Listeria monocytogenes
EAMERSCT1EJ2; International audience; No abstract
Listeria monocytogenes, a down-to-earth pathogen
International audience; Listeria monocytogenes is the causative agent of the food-borne life threatening disease listeriosis. This pathogenic bacterium received much attention in the endeavor of deciphering the cellular mechanisms that underlie the onset of infection and its ability to adapt to the food processing environment. Although information is available on the presence of L. monocytogenes in many environmental niches including soil, water, plants, foodstuff and animals, understanding the ecology of L. monocytogenes in outdoor environments has received less attention. Soil is an environmental niche of pivotal importance in the transmission of this bacterium to plants and animals. Soil…
Le rôle du système agr chez Listeria monocytogènes
National audience; La capacité des bactéries à développer un biofilm constitue une des principales difficultés quant à la maîtrise de la contamination : ces structures en augmentent les risques et rendent notamment les micro-organismes plus résistants aux désinfectants. Des études ont pu démontrer le rôle du système de communication agr dans la formation de biofilms de L. monocytogenes, agent pathogène particulièrement recherché en industrie agro-alimentaire.
Assessment of the roles of LuxS, Autoinducer 2 and its precursor S-Ribosyl Homocysteine, in cell attachment during biofilm formation by Listeria monocytogenes EGD-e
No abstract
Short-term impact of biogas digestates on soils microbial communities
International audience; Anaerobic digestion of organic waste is considered a key process to produce renewable energy to meet the growing sustainable energy demand. Digestates can be used in agriculture as soil amendments and improve crop yields. However, their use at large scale in agricultural fields still requires to prove their innocuity on soil biota, especially on microorganisms that play important roles in soil ecosystem. Here, we designed a microcosm experiment to compare the short-term (42-days) effects of four different digestates (derived from cattle manure, energy crop, food residues or slurry with bio-waste) on the soil microbial communities. Each digestate was applied on three …
Bactéries environnementales : intérêt biotechnologique et enjeux sanitaires
Diplôme : HDR
Influence des conditions environnementales (statique versus dynamique) sur le développement en biofilm de Listeria monocytogenes EGD-e. Developpement en biofilm et expression d'Agr de Listeria monocytogenes
National audience
Indigenous Listeria spp. in biowastes and during composting
International audience
Le système [i]agr[/i] de [i]Listeria monocytogenes[/i] et la croissance en biofilm : revisite des notions de communication, quorum sensing et auto-induction
Communication orale, résumé; Parmi les mécanismes cellulaires qui participent à la mise en place des biofilms, l’échange de molécules de communication apparaît nécessaire chez de nombreux microorganismes. Listeria monocytogenes est une bactérie ubiquiste responsable de pathologies sévères chez des populations à risque (immunodéprimés, femmes enceintes, personnes âgées) en cas de consommation d’aliments contaminés. Nous avons récemment démontré que le système de communication agr est impliqué au cours de la croissance sessile de cette bactérie (Rieu et al., 2007). Par ailleurs, l’expression du système agr était hétérogène pendant la croissance en biofilm (Rieu et al., 2008) et localisée à sa…
La biodiversité de la microflore du sol prévient l’implantation de pathogènes alimentaires dans les sols agricoles
Pôle EA MERS; National audience; La présence dans les agroécosystèmes de bactéries pathogènes pouvant être transmises à l’homme représente un risque sanitaire pour l’ensemble de la filière de production des aliments. Le sol est en effet une zone de transfert potentiel de ces agents pathogènes vers les matières premières agricoles, puis vers les produits agroalimentaires, la plupart du temps dans des produits animaux, mais aussi via des produits végétaux également.
Differential expression of the agr locus of Listeria monocytogenes EGD-e
International audience
LisRK is required for optimal fitness ofListeria monocytogenesin soil
ABSTRACTListeria monocytogenes is a food-borne pathogen responsible for the disease listeriosis. It is ubiquitously found in the environment and soil is one of its natural habitats. Listeria monocytogenes is highly capable of coping with various stressful conditions. We hypothesized that stress-responsive two-component systems such as LisRK might contribute to the adaptation of L. monocytogenes to the soil environment. Indeed, investigations of the population dynamics of wild-type and mutant strains suggest an important role of LisRK for optimal fitness of L. monocytogenes in sterile soil. Results from non-sterile soil showed that the parental strain was capable of surviving longer than mut…
Development of phylogroup-specific taqman real-time assays for identification of members of klebsiella pneumoniae complex
International audience; • Klebsiella Klebsiellapneumoniae pneumoniae pneumoniae (Kp )represents agrowinggrowing public publichealth concern due to the emergence emergence of multidrug multidrug multidrug resistance resistance and andhypervirulence hypervirulence . • The Kp species complex complex includes includes 7closely related related phylogroups :Klebsiella Klebsiellapneumoniae (phylogroup (phylogroup Kp 1), K.quasipneumoniae quasipneumoniae quasipneumoniae subsp .quasipneumoniae quasipneumoniae (Kp 2)and subsp .similipneumoniae similipneumoniae similipneumoniae similipneumoniae similipneumoniae (Kp 4), K.variicola variicola subsp .variicola variicola variicola(Kp 3)and subsp .tropical…
Agr system of Listeria monocytogenes EGD-e: role in adherence and differential expression pattern.
ABSTRACT In this study, we investigated the agrBDCA operon in the pathogenic bacterium Listeria monocytogenes EGD-e. In-frame deletion of agrA and agrD resulted in an altered adherence and biofilm formation on abiotic surfaces, suggesting the involvement of the agr system of L. monocytogenes during the early stages of biofilm formation. Real-time PCR experiments indicated that the transcript levels of agrBDCA depended on the stage of biofilm development, since the levels were lower after the initial attachment period than during biofilm growth, whereas transcription during planktonic growth was not growth phase dependent. The mRNA quantification data also suggested that the agr system was a…
Communication and Autoinduction in the species Listeria monocytogenes
International audience; In order to withstand changes in their environment, bacteria have evolved mechanisms to sense the surrounding environment, integrate these signals and adapt their physiology to thrive under fluctuating conditions. Among these mechanisms, the ability of bacteria to exchange information between cells has become a dynamic field of interest for microbiologists over the past four decades. First described by Nelson et al.,1 this phenomenon often referred as either cell-cell communication, Quorum Sensing and/or AutoInduction involves the synthesis of small signal molecules called autoinducers. These signal molecules may be sensed by the bacterial population in the vicinity …
Involvement of communication systems during the sessile growth of Listeria monocytogenes
International audience
Evidence of autoinduction heterogeneity via expression of the agr system of listeria monocytogenes at the single-cell level
ABSTRACT To investigate if the primary function of the Agr system of Listeria monocytogenes is to monitor cell density, we followed Agr expression in batch cultures, in which the autoinducer concentration was uniform, and in biofilms. Expression was heterogeneous, suggesting that the primary function of Agr is not to monitor population density.
Listeria monocytogenes EGD-e biofilms: no mushrooms but a network of knitted chains.
ABSTRACT Listeria monocytogenes is a food pathogen that can attach on most of the surfaces encountered in the food industry. Biofilms are three-dimensional microbial structures that facilitate the persistence of pathogens on surfaces, their resistance toward antimicrobials, and the final contamination of processed goods. So far, little is known about the structural dynamics of L. monocytogenes biofilm formation and its regulation. The aims of this study were, by combining genetics and time-lapse laser-scanning confocal microscopy (LSCM), (i) to characterize the structural dynamics of L. monocytogenes EGD-e sessile growth in two nutritional environments (with or without a nutrient flow), and…
The σ B -dependent regulatory sRNA Rli47 represses isoleucine biosynthesis in Listeria monocytogenes through a direct interaction with the ilvA transcript
International audience; The facultative intracellular pathogen Listeria monocytogenes can persist and grow in a diverse range of environmental conditions, both outside and within its mammalian host. The alternative sigma factor Sigma B (sigma(B)) plays an important role in this adaptability and is critical for the transition into the host. While some of the functions of the sigma(B) regulon in facilitating this transition are understood the role of sigma(B)-dependent small regulatory RNAs (sRNAs) remain poorly characterized. In this study, we focused on elucidating the function of Rli47, a sigma(B)-dependent sRNA that is highly induced in the intestine and in macrophages. Using a combinatio…
Nation-wide study of the occurrence of Listeria monocytogenes in French soils using culture-based and molecular detection methods
Identifiant HAL : hal-01120618; International audience; Soil is a potential reservoir of human pathogens and a possible source of contamination of animals, crops and water. In order to study the distribution of Listeria monocytogenes in French soils, a real-time PCR TaqMan assay targeting the phosphoribosylpyrophosphate synthetase (prs) gene of L. monocytogenes was developed for the specific detection and quantification of this bacterium within a collection of 1315 soil DNAs originated from the French Soil Quality Monitoring Network. The prs real-time PCR TaqMan assay was specific for L. monocytogenes and could quantify accurately down to 104L. monocytogenes per gram of dry soil. Among the …
Listeria monocytogenes
Prod 2017-266 équipe EA UB BIOME; National audience; résumé du livre : Pour garantir et maîtriser la sécurité microbiologique des aliments et prévenir les crises sanitaires alimentaires, la connaissance et la surveillance des microorganismes pathogènes depuis la production primaire jusqu’à la distribution des denrées alimentaires en passant par la transformation, sont indispensables. Cet ouvrage de référence traite des dangers microbiologiques alimentaires majeurs (microorganismes infectieux ou toxines d’origine microbienne) et des risques associés pour l’Homme. Illustré de nombreux schémas et tableaux de synthèse, il fait un point complet sur les notions fondamentales de microbiologie géné…
Investigation of rhizosphere as a niche for Listeria monocytogenes
EAUBAGROSUPBIOME; International audience; No abstract
Natural soil reservoirs for human pathogenic and fecal indicator bacteria
Prod ? EA UB INRA BIOME; International audience; résumé du livre : Environmental microbiology, the study of the roles that microbes play in all planetary environments, is one of the most important areas of scientific research. The The Manual of Environmental Microbiology, Fourth Edition, provides comprehensive coverage of this critical and growing field. Thoroughly updated and revised, the Manuall is the definitive reference for information on microbes in air, water, and soil and their impact on human health and welfare. Written in accessible, clear prose, the manual covers four broad areas: general methodologies, environmental public health microbiology, microbial ecology, and biodegradati…
Distribution and characteristics of Listeria monocytogenes isolates from surface waters of the South Nation River Watershed, Ontario, Canada
ABSTRACT Listeria monocytogenes is a facultative intracellular pathogen thought to be widely distributed in the environment. We investigated the prevalence and characteristics of L. monocytogenes isolates from surface waters derived from catchments within the South Nation River watershed (Ontario, Canada). This watershed is dominated by urban and rural development, livestock and crop production, and wildlife habitats. From June to November 2005, a total of 314 surface water samples were collected biweekly from 22 discrete sampling sites characterized by various upstream land uses. Presumptive Listeria spp. were isolated using a selective enrichment and isolation procedure, and 75 L. monocyt…
Microbial diversity and structure are drivers of the biological barrier effect against Listeria monocytogenes in soil
International audience; Understanding the ecology of pathogenic organisms is important in order to monitor their transmission in the environment and the related health hazards. We investigated the relationship between soil microbial diversity and the barrier effect against Listeria monocytogenes invasion. By using a dilution-to-extinction approach, we analysed the consequence of eroding microbial diversity on L. monocytogenes population dynamics under standardised conditions of abiotic parameters and microbial abundance in soil microcosms. We demonstrated that highly diverse soil microbial communities act as a biological barrier against L. monocytogenes invasion and that phylogenetic compos…
Interactions in dual species biofilms between Listeria monocytogenes EGD-e and several strains of Staphylococcus aureus.
International audience; Six environmental isolates of Staphylococcus aureus and one collection strain were investigated for their ability to form monospecies biofilms and dual species biofilms with Listeria monocytogenes EGD-e on stainless steel coupons. All isolates were able to grow as biofilms but their ability to form monospecies biofilms differed. The population of L. monocytogenes EGD-e in dual species biofilms was not affected by the presence of S. aureus isolates except for strain CIP 53.156. The effect of L. monocytogenes EGD-e on the population of S. aureus was strain dependent: S. aureus population either increased or decreased or was not affected in the presence of L. monocytoge…
Single cell analysis evidence Heterogeneous expression of the Agr communication system of Listeria monocytogenes
http://prodinra.inra.fr/record/271694EAMERSCT3 EJ3; Communication, often referred as Quorum Sensing, is involved in the adaptation of most bacteria to their environment. In the genus Listeria, the agr communication system affects the biology of Listeria monocytogenes during saprophytic life (biofilm formation) and during infection. In the present study, we investigated in situ agr expression of six isolates of L. monocytogenes by combining gfp reporters, flow cytometry and fluorescent microscopy during plantonic growth and dynamic flow cell biofilm set ups. Unexpectedly, during growth in TSB homogenised liquid cultures incubated at 25°C, while auto-inducer concentration was uniform, statist…
σB and agrA regulation in Listeria monocytogenes: Effect on survival in soil/rhizosphere under biotic and abiotic conditions
International audience; Abstract -250 words Backgrounds: Listeria monocytogenes is the agent of listeriosis, a life-threatening condition in at-risk people. Complex transmission routes between outdoor environments and the food chain result in foodstuff contamination. Sensing of environmental changes can trigger regulation of gene expression, allowing bacteria to adapt their physiology and survive. The Agr cell-cell communication system transcription regulator AgrA is triggered during several environmental conditions including soil, an important reservoir of L. monocytogenes. The RNA polymerase σB factor aids survival in several stress conditions and may be required for L. monocytogenes surv…
Sigma B-dependent regulatory sRNA Rli47 represses isoleucine biosynthesis in Listeria monocytogenes
International audience; Background: Small regulatory RNAs (sRNAs) can influence cellular processes by targeting mRNAs, leading to translation inhibition and mRNA degradation. Previous studies have shown that the σB-dependent sRNA Rli47 is induced in L. monocytogenes stationary phase cells, in the intestinal lumen and macrophages, but its function has remained elusive. Objectives: This study aimed to comprehensively characterize Rli47 and determine its biological function in L. monocytogenes. Methods: Besides in silico predictions, several molecular approaches such as DMS-MaPseq, northern blotting, EMSAs, RTqPCR, RNA-seq and phenotypic experiments were applied. Results: Rli47 was shown to ba…
Exploring the diversity of listeria monocytogenes biofilm architecture by high-throughput confocal laser scanning microscopy and the predominance of the honeycomb-like morphotype
ABSTRACT Listeria monocytogenes is involved in food-borne illness with a high mortality rate. The persistence of the pathogen along the food chain can be associated with its ability to form biofilms on inert surfaces. While most of the phenotypes associated with biofilms are related to their spatial organization, most published data comparing biofilm formation by L. monocytogenes isolates are based on the quantitative crystal violet assay, which does not give access to structural information. Using a high-throughput confocal-imaging approach, the aim of this work was to decipher the structural diversity of biofilms formed by 96 L. monocytogenes strains isolated from various environments. Pr…
Techniques d'études de la formation des biofilms. Retour d'expérience issu du domaine "agro-alimentaire" et du domaine "environnemental"
No abstract
Survival of Listeria monocytogenes in Soil Requires AgrA-Mediated Regulation
ABSTRACT In a recent paper, we demonstrated that inactivation of the Agr system affects the patterns of survival of Listeria monocytogenes (A.-L. Vivant, D. Garmyn, L. Gal, and P. Piveteau, Front Cell Infect Microbiol 4:160, http://dx.doi.org/10.3389/fcimb.2014.00160 ). In this study, we investigated whether the Agr-mediated response is triggered during adaptation in soil, and we compared survival patterns in a set of 10 soils. The fate of the parental strain L. monocytogenes L9 (a rifampin-resistant mutant of L. monocytogenes EGD-e) and that of a Δ agrA deletion mutant were compared in a collection of 10 soil microcosms. The Δ agrA mutant displayed significantly reduced survival in these b…
Investigation of the roles of AgrA and σB regulators in Listeria monocytogenes adaptation to roots and soil
ABSTRACT Little is known about the regulatory mechanisms that ensure the survival of the food-borne bacterial pathogen Listeria monocytogenes in the telluric environment and on roots. Earlier studies have suggested a regulatory overlap between the Agr cell–cell communication system and the general stress response regulator σB. Here, we investigated the contribution of these two systems to root colonisation and survival in sterilised and biotic soil. The ability to colonise the roots of the grass Festuca arundinacea was significantly compromised in the double mutant (∆agrA∆sigB). In sterile soil at 25°C, a significant defect was observed in the double mutant, suggesting some synergy between …
Listeria monocytogenes agr system: Quorum sensing, or maybe not?
International audience
Le système Agr de Listeria monocytogenes : revisite des notions de Quorum Sensing et auto-induction par cytométrie en flux
National audience
Listeria monocytogenes Differential Transcriptome Analysis Reveals Temperature-Dependent Agr Regulation and Suggests Overlaps with Other Regulons
Listeria monocytogenes is a ubiquitous, opportunistic pathogenic organism. Environmental adaptation requires constant regulation of gene expression. Among transcriptional regulators, AgrA is part of an auto-induction system. Temperature is an environmental cue critical for in vivo adaptation. In order to investigate how temperature may affect AgrA-dependent transcription, we compared the transcriptomes of the parental strain L. monocytogenes EGD-e and its Delta agrA mutant at the saprophytic temperature of 25 degrees C and in vivo temperature of 37 degrees C. Variations of transcriptome were higher at 37 degrees C than at 25 degrees C. Results suggested that AgrA may be involved in the regu…
Sensitivity to acetic acid, ability to colonize abiotic surfaces and virulence potential of Listeria monocytogenes EGD-e after incubation on parsley leaves.
International audience; Abstract Aim: To investigate how the survival of Listeria monocytogenes on parsley leaves may affect its ability to sustain process-related harsh conditions and its virulence. Methods and Results: Parsley seedlings were spot inoculated with stationary phase cells of L. monocytogenes EGD-e and incubated for 15 days. Each day, bacterial cells were harvested and enumerated, and their ability to survive acetic acid challenge (90 min, pH 4.0), to colonize abiotic surfaces and to grow as biofilms was assessed. After a 3-log decrease over the first 48 h, the population stabilized to about 10(6) CFU g(-1) until the sixth day. After the sixth day, L. monocytogenes was no long…
Long-term survival of pathogenic and sanitation indicator bacteria in experimental biowaste composts
ABSTRACT For economic, agricultural, and environmental reasons, composting is frequently used for organic waste recycling. One approach to limiting the potential risk from bacterial food-borne illnesses is to ensure that soil amendments and organic fertilizers are disinfected. However, more knowledge concerning the microbiological safety of composted substrates other than sludge and manure is necessary. Experimental in-vessel biowaste composts were used to study the survival of seeded Listeria monocytogenes , Salmonella enterica subsp. enterica serotype Enteritidis, and Escherichia coli . Four organic waste mixtures, containing various proportions of paper and cardboard, fruits and vegetabl…
Biofilms structural diversity of 100 strains of the food-borne pathogen Listeria monocytogenes
International audience
Changes in gene expression during adaptation of Listeria monocytogenes to the soil environment
project SEST 009; International audience; Listeria monocytogenes is a ubiquitous opportunistic pathogen responsible for listeriosis. In order to study the processes underlying its ability to adapt to the soil environment, whole-genome arrays were used to analyse transcriptome modifications 15 minutes, 30 minutes and 18 h after inoculation of L. monocytogenes EGD-e in soil extracts. Growth was observed within the first day of incubation and large numbers were still detected in soil extract and soil microcosms one year after the start of the experiment. Major transcriptional reprofiling was observed. Nutrient acquisition mechanisms (phosphoenolpyruvate-dependent phosphotransferase systems and…
The σB-dependent regulatory sRNA Rli47 represses isoleucine biosynthesis in Listeria monocytogenes through a direct interaction with the ilvA transcript
The facultative intracellular pathogen Listeria monocytogenes can persist and grow in a diverse range of environmental conditions, both outside and within its mammalian host. The alternative sigma factor Sigma B (σB) plays an important role in this adaptability and is critical for the transition into the host. While some of the functions of the σB regulon in facilitating this transition are understood the role of σB-dependent small regulatory RNAs (sRNAs) remain poorly characterized. In this study, we focused on elucidating the function of Rli47, a σB-dependent sRNA that is highly induced in the intestine and in macrophages. Using a combination of in silico and in vivo approaches, a binding…