2018

DomaineduHaut-Carré,U.Bordeaux24&25septembre

MondaySeptemberthe24th8h30 WELCOME9h15 INTRODUCTION9h30 ECOLOGY&EVOLUTION @Agora 9h30 PatrickFORTERRE(40min+5)InstitutPasteur,ParisDrawingviral"treesoflife"thatraiseintriguingevolutionaryquestions

10h15SylvainGANDON(15min+5)CEFE,CNRSMontpellierPathogensdrivetheevolutionofhostresistancediversity10h35MarianneDEPAEPE(15min+5)MICALIS,INRA,Jouy-En-JosasPhage-bacteriacoevolutioninthemousegut10h55COFFEE-BREAK11h20NicolasGINET(15min+5)Laboratoire de Chimie Bactérienne, UMR7283, CentreNational de la Recherche ScientifiqueUMR7283,AMUSmuggling of magnetotaxis-related genes by a temperate bacteriophage inMagnetospirillummagneticumAMB-111h40CamilleD’HUMIERES(15min+5)UniversitéParisDiderot-Paris7SorbonneParisCité,Paris-MicrobialEvolutionaryGenomics,InstitutPasteurdeParisImpactofantibioticsonhumangutphageome12hAdélaïdeRENARD(15min+5)EquipeBactériesetrisquematerno-fœtalUMR1282InfectiologieetSantépublique,ParisFunctional impact of A-prophage with Streptococcus agalactiae adaptation facingLactobacillusfromthevaginalflora.12h20AraianeBIZE(15min+5)Irstea,URHBAN,1ruePierre-GillesdeGennes,F-92761Antony,France.K-merapproachesprovidevaluable insight intomobilomeevolution in thedomainArchaea12h40LUNCH&POSTERSESSION @BadianeRoom&Gallery

14h30AstridWAHL(15min+5)Laboratoire de Chimie Bactérienne, UMR7283, CentreNational de la Recherche ScientifiqueUMR7283,AMUControlandmaintenanceofprophagesinSalmonellaenterica14h50StéphaneCHAILLOU(15min+5)InstitutMICALIS,INRA,AgroPArisTech,UniversitéParis-Saclay.Jouy-en-Josas.Characterizationandmodellingofphagespopulationstructureinsmearwashed-rindcheeseundervariousenvironmentalripeningconditions.

15h10PHAGE-HOSTMOLECULARINTERACTIONS @Agora

15h10 JennyferMAHONY(40min+5)UCCCork“Somelikeitsweet”–Hostrecognitionbyphagesoflacticacidbacteria15h55COFFEE-BREAK16h30LiaMARQUESGODINHO(15min+5)DépartementdeVirologie,InstitutdeBiologieIntégrativedelaCelluleCNRSUMR9198FunctionofGroELduringbacteriophageinfectionofBacillussubtilis16h50MathieuDEJODE(15min+5)InstitutPasteurUniversitéSorbonneParisCité(USPC)A single phage protein efficiently disrupts the host physiology by targeting sigmafactors

17h10IsabelleBERTRAND(15min+5)Laboratoire de Chimie Physique et Microbiologie pour les Matériaux et l’EnvironnementCentreNationaldelaRechercheScientifique:UMR7564,UniversitédeLorraineEffectsofheatandchlorineon the inactivationandphysico-chemicalpropertiesofbacteriophageMS217h30Marie-AgnèsPETIT(15min+5)MICrobiologie de l’ALImentation au Service de la Santé humaine, Institut National de laRechercheAgronomique:UMR1319,AgroParisTechVirulent coliphages isolated from 1 year-old children gut microbiota are sub-dominantcomparedtotemperates,buthighlyinfectious17H50END20h00DINNER @Citéduvin,Bordeaux(accessbytram)

TuesdaySeptemberthe25th8h30WELCOME9h00 THERAPYANDBIOTECHNOLOGYAPPLICATIONS @Agora9h00JoanaAZEREDO(40min+5)UniversityMinho,BragaPhages/biofilminteraction:strategiestoimprovephageefficacyagainstinfectiousbiofilms9h45MartineBOCCARA(15min+5)InstitutLangevin,ESPCIParis,PSLResearchUniversity,1rueJussieu,75005Paris,France.Virusesandmembranesvesiclesproducedbydenitrifyingbacteria10h05 FernandoCLAVIJO(15min+5)LaboratoiredeChimieBactérienne,UMR7283CentreNationalde laRechercheScientifique -AixMarseilleUniversité;InstitutdeRechercheenHorticultureetSemences,INRA,UMR1345;BiolineAgrosciencesAssessingphagetherapyagainsttheplantpestXylellafastidiosa10h25 PilarGARCIA(15min+5)InstitutodeProductosLácteosdeAsturias,IPLA-CSIC.Spain.SpanishNetworkofBacteriophagesandTransducterElements(FAGOMA)10h45COFFEE-BREAK11h15GilbertVERBEKEN(40min+5)HopitaldelaReineAstrid,Bruxelles“BacteriophageTherapy:FactorFictioninWesternMedicine”12hRaphaëlleDELATTRE(15min+5)Groupe IBBA, unité BMGE, Institut Pasteur/IAME, Inserm, Paris/service d’anesthésie-réanimationHôpitalBeaujon,ClichyPreliminary studies on pharmacokinetics and pharmacodynamics of virulentbacteriophages treating Pseudomonas aeruginosa and Escherichia coli lunginfections.12h20Frédéric-AntoineDauchy(15min+5)CHUPellegrinBordeauxPhagothérapiedanslesinfectionsostéo-articulaires12h40 LUNCH14h30Assembléegénérale

ECOLOGY & EVOLUTION

Drawingviral"treesoflife"thatraiseintriguingevolutionaryquestionsPatrickForterre1,2,AnthonyWoo1,JulienGuglialmini3,AlexisCrisculo3,VioletteDaCunha1,2andMorganGaia1.1InstitutPasteur,DépartementdeMicrobiologie,25rueduDocteurRoux,Paris,France.2 Institut de Biologie Intégrative de la cellule,Département deMicrobiologie,Université Paris-Saclay,Orsay,France.3InstitutPasteurCenterofBioinformatics,BiostatisticsandIntegrativeBiology,25rueduDocteurRoux,Paris,France.

Isitpossibletoplacevirusesinuniversaltreesoflife?Iwillbrieflydiscussthisquestionand its corollaries: are viruses living? What is a virus? How and when virusesoriginated?Mypersonalviewisthatvirusesare«here,thereandeverywhere»inthetreeoflifebecausetheyinfectallcellularorganisms.However,virusesarepolyphyleticandcannotbeeasilytracedtotheirancestors.TwomajorlineagesofDNAviruseshavebeen defined that are present in the three cellular domains, Archaea, Bacteria andEukarya: theAdenovirus/PRD1 lineage and theHK97 lineages, both definedby theirmajorcapsidproteinsandpackagingATPases.Iwillpresentourpreliminaryattemptsto decipher the history of these lineages through phylogenetic analyses. We haveobtaineduniversaltreesoflifeofthePRD1andHK97«superlineages»thatareatoddwith either the classical Woese’s tree of cellular domains or the fashionable (butprobably wrong) 2D (eocyte) tree. This raises exciting questions about the role ofviruses in the evolution of the three cellular domains. Notably,we have obtained arobust tree of the “Adenovirus/PRD1 sub-lineage” corresponding to theNucleoCytoplasmicLargeDNAViruses(NCLDV).Moreover,NCLDVcanbeplacedinthetreeoflifebasedontheirRNApolymerases.OurresultsindicatethatNCLDVhaveco-evolved with proto-eukaryotes, indicating that they can have played a role in theformationofmoderneucaryotes.

Type=oral

LesphagesgouvernentladynamiquedeladiversitéderésistancedesbactériesS.Gandon,H.Chabas,E.OrtegaAbboud,A.NicotCEFE,Montpellier

Ladiversitédelarésistanceauxpathogènesdanslespopulationshôtespeutavoirdeseffets importants sur les épidémies. Cependant, l’impact des pathogènes sur cettediversité est bien moins étudié. Nous avons développé un système expérimentalpermettant de suivre l’influence de l’exposition aux phages sur la dynamiquedémographiqueetsurladiversitéderésistanceCRISPRchezlesbactérieshôtes.Nousmontrons qu’en l’absence de phages, les populations perdent très rapidement ladiversitéderésistance.L’expositionàunepopulationdephagesnondiversifiésmèneégalement à une perte de cette diversité à court terme, mais cette diversité estrestaurée au bout de quelques jours suite à l’acquisition de nouvelles résistances.L’exposition à une population de phages diversifiésmaintient la diversité initiale derésistanceetengendre rapidementunedifférentiation importanteentrepopulationshôtes. Ces résultats illustrent l’impact de la pression de sélection imposée par lesphages sur ladémographieet l’évolutionde leurshôtes. Ilsdémontrentaussique lacoévolution antagoniste peut jouer un rôle majeur dans le maintien de la diversitélocaleetdansladifférentiationentrepopulations.

Type: : posterMots-Clés : CRISPR,résistance,diversité

Phage-bacteriacoevolutioninthemousegutMarianneDePaepe,JeffreyCornuault,Marie-AgnèsPetit,ElisabethMoncautMICALISInstitutnationaldelarechercheagronomique(INRA),UMR1319

Bacteriophagesareverynumerousinthegutmicrobiota,butsofarthereislittleproofthat they are important determinants of bacterial mortality in this environment. Inmousestudies,phage-mediatedbacterialmortalityingenerallylimitedtoafractionofthesusceptiblebacteria, suggesting thatphysiologycomplicatesphagemultiplicationintheintestine.

We followed in gnotoxenic mice the population dynamics of a dominant gutmicrobiotaspecies,Roseburia intestinalisL1-82,and itsphageShimadzu.Shimadzu isinitially aR. intestinalis prophage, that systematically evolves towards ultravirulence(the ability to infect a lysogen) in this simplified gut ecosystem, thanks to theacquisitionofmutations in the lysis/lysogeny regulatory region.Ultravirulentphagesalsosystematicallyacquiremutationsintheirtailfiber,thankstoadiversity-generatingretroelement(DGR).Inmice,phageinfectionkilledthemajorityofsusceptiblebacteriaina fewdays, leading to the fastselectionof resistantbacteria,mainly throughnewspaceracquisitioninaCRISPRarray.Intriguingly,thelargemajorityofvirionsisolatedfrommouse fecesareunableto infectbacteria invitro. Inaddition, invitro,bacteriainfectedbyphagesseemunabletoacquirenewspacersandbecomephageresistant,suggestingthatdifferentmechanismsareinvolvedinvitroandinthemousegut.

Altogether,theseresultsthatforthegutsymbiontR.intestinalis,bothphageinfectionand bacterial adaptation are more efficient in vivo,which suggests that phage areimportantkillerofthisbacterium.

Type: : oralKey-words : microbioteintestinal;CRISPR;prophage

Smuggling of magnetotaxis-related genes by a temperate bacteriophage inMagnetospirillummagneticumAMB-1NicolasGinet1,2,MireilleAnsaldi1,21LaboratoiredeChimieBactérienne,UMR7283,CentreNationaldelaRechercheScientifiqueCNRS,UMR72832AixMarseilleUniversité

Horizontalgenetransfers(HGT)enablerapidacquisitionandevolutionofnewabilitiescoming from other - sometimes distantly related - organisms. Among the variousvectors of HGT in bacteria, viruses (or bacteriophages) play a prominent role bytransferringgeneticmaterialbetweenindividualseitherbytransduction(specializedorgeneralized)or,inthecaseoftemperatephages,bylysogenization(stableassociationofthephagegenome-thentermedprophage-withthehost's).Lysogenymaybenefittothehost,forinstancebyraisingpathogenicityofvibriotowardshumans(cytotoxinCtxAB responsible for cholera is encoded within the genome of the temperatefilamentousphageCTX).Theprophage is thenpassively replicatedwith thebacterialgenome until changes in environmental conditions induce its excision and theresumingofthelyticcycle;alternativelyitmaybedefinitivelydomesticatedwithinthebacterial genome throughmutations, deletions or genetic rearrangements. Bacterialmagnetotaxis-theabilitytonavigatealongthegeomagneticfieldlines-isenabledbya series of specific genes shared by magnetotactic bacteria (MTB). ThesemicroorganismsformapolyphyleticgroupandseverallinesofevidencessuggestthatHGT events participated to this diversity. We present here evidences that theprophageFRODOfoundinthegenomeofthemodelMTBbacteriumMagnetospirillummagneticum AMB-1 can excise from its host's genome and form viral particles thattransport the magnetotaxis-related genes clustered in the Magnetotactic Isletspecificallybelongingtothisstrain.

Type: : oralKey-words : Prophage;horizontalgenetransfer;magnetotacticbacteria

ImpactofantibioticsonhumangutphageomeCamilleD'humieres1,2,3,4,5,MarieTouchon4,5,SaraDion3,ErickDenamur6,3,7,EduardoRocha4,5,PrediresGroup1UniversitéParisDiderot-Paris7SorbonneParisCité,F-75018Paris,-France2AP-HP,HôpitauxUniversitairesParisNordVal-de-Seine,LaboratoiredeBacteriologieHôpitalBichat-ClaudeBernard46RueHenriHuchard-France3INSERM,IAME,UMR11374CNRS,UMR35255MicrobialEvolutionaryGenomics,InstitutPasteurdeParis6AP-HP,HôpitauxUniversitairesParisNordVal-de-Seine7UniversitéParisDiderot-Paris7

The microbiota of the human gut sometimes called «the second brain» is a verycomplex and rich community composed of bacteria, archaea, protozoa, fungi andviruses,coexistinginequilibrium.Thestudyofthedynamicsandstabilityofthehumangutmicrobiotaisessentialtounderstanditsroleindisease,diagnosis,treatmentandprevention. Bacteria and their viruses (bacteriophages) are the most importantmembersofthiscommunity.Intherecentyearsbacteriophagesfromguthavestartedtoreceiveattention,contributionofphagestointestinalmicrobiotaecologyandtheireffectonhumanhostarejustbeginningtobehighlighted.In thiswork,we studied the impactof antibioticson thehumangutphageome.Wecompared two third generation cephalosporins with distinct pharmacokineticcharacteristics, ceftriaxone having higher biliary elimination than cefotaxime. Aprospective, monocenter, open, randomized clinical trial in 22 healthy volunteerstreatedbythe intravenousrouteeitherwithceftriaxone(1g/24hours)orcefotaxime(1g/8 hours) for 3 dayswas performed as part of a project funded by the “AgenceNationalede laRecherche”.Thisallowedthe follow-upof individuals forsixmonths,before,duringandaftertreatment.Preliminaryresultsshowedadifferencebetweenthetwocephalosporinsonhumanphageomediversity.Healthyvolunteerstreatedbyceftriaxone (higher biliary elimination) showed stronger perturbations of their gutphageomethanthevolunteerstreatedwithcefotaxime,withadrasticdecreaseofthepredominantphagecontigsobservedjustaftertreatmentandareturntothebaselineleveltendaysaftertreatment.Moreanalyseswillbeperformedtoidentifythedriversoftheobservedphageomedisruption.

Type: : oralThématiques : EcologieetévolutionMots-Clés : Instestinalmicrobiotephageomeantibiotic

Functional impact of A-prophage with Streptococcus agalactiae adaptation facingLactobacillusfromthevaginalflora.AdélaïdeRenard 1,PaulineCantin 1, SandraDosSantosBorges 2,MarionLacasse 1,Anne-SophieValentin1,LaurentMereghetti1,RolandQuentin1,NathalieVanDerMee-Marquet1

1EquipeBactériesetrisquematerno-fœtalUMR1282InfectiologieetSantépublique2ServicedeBactériologieCHUTrousseauAPHP

Context:Streptococcusagalactiae(GBS)hasbecomesincethe1960stheleadingcauseof neonatal infection in industrialised countries. GBS neonatal infections aremostlyassociatedwiththeclonalcomplexCC17.GBSiscolonizingthevaginaltract in1/3ofpregnant women. The CC17-GBS neonatal infections mainly occur during deliveryfollowingtransmissionofGBStoneonatesfrommotherscarryingGBSintheirvagina.Vaginalmicrobiota,dominatedbyLactobacilluscrispatus,preventvaginalcolonizationbypathogens,includingGBS,andhasbeensuggestedplayingaprotectiveroleagainstGBS neonatal infection. Using whole genome sequencing of 14 GBS strainsrepresentativeofthespecies,weidentified22prophagesclusteredinto6groupsAtoF. CC17 GBS strains 10avor10o n10e for neonatal infections frequently carry A-prophages inserted near bacterial genes involved in adaptation, stress resistance orvirulence, and carrying genes associated to defense systems, adaptive response andputativevirulencefactor.Ourhypothesis:prophageAmayhaveapositive10avor10onGBSadaptationfacingLactobacilli,andthusmay10avorGBScolonization intothevaginaandconsequentGBSneonatalinfections.Strategy and results: First, we have constructed by deletion or addition of a A-prophage, one couple of isogenic GBS strains differing by their prophage content.Second,wehavestudiedandcomparedtheisogenicstrainsregarding(1)theirgrowthcurveswithorwithoutthepresenceofLactobacillusculturesupernatant,and(2)theircapacity to form biofilm. Our findings showed significantly different results forprophage-freeandlysogenicGBS.Wefirstdemonstratedmajordifferencesregarding(1)themaximalgrowthofstrains(+15%forthelysogenicstrainwhencomparedwiththe prophage-free strain), (2) the growth rate of strains (in the presence ofLactobacillussupernatant,thegenerationtimefortheprophagefreestrainwas+43%to +95% compared with the lysogenic strain), and (3) biofilm formation (BiofilmFormation Index twice as important for lysogenic strain when compared withprophage-freeGBS).Conclusion and perspectives. Our data suggest a functional impact of A-prophagecarried by GBS strains of CC17. Further investigations are currently performed toinvestigatethemolecularmechanismsassociatedwiththephenotypesobserved.

Type: : oralKey-words : Streptococcusagalactiae;temperatephages.

K-merapproachesprovidevaluable insight intomobilomeevolution in thedomainArchaeaArianeBize1,VioletteDaCunha2-3,CédricMidoux1,SophieSchbath4,PatrickForterre1-21Irstea,URHBAN,1ruePierre-GillesdeGennes,F-92761Antony,France.2Institut Pasteur, Unité de Biologie Moléculaire du Gène chez les Extrêmophiles, Département deMicrobiologie,25rueduDocteurRoux,75015Paris,France.3Institute for IntegrativeBiologyof theCell (I2BC),CEA,CNRS,Univ.Paris-Sud,UniversitéParis-Saclay,Gif-sur-Yvettecedex,France4INRA,UR1404Mathematique InformatiqueAppliquéesduGénomeà l'Environnement, Jouy-en-Josas,FranceKmerapproacheshavegreatlydevelopedinrecentyears,largelydrivenbytheadventofnext-generationsequencing.Theirspeedandautomatism,sincetheyareannotationindependent, are major advantages. The interest of applying k-mer approaches tostudythemobilomeisonlystartingtobeexplored,withmanyeffortsorientedtowardsviral metagenomics [1, 2]. We evaluated the potential of applying simple k-merapproachestounderstandtheevolutionaryhistoryofmobileelementsbyfocusingonviruses and plasmids from the domain Archaea, which includes specific and well-definedextrachromosomalelementfamilies[3].We selected more than 590 cell, virus and plasmid genomes, originating from 11distinct orders of archaeal cells. We subsequently implemented multivariate andstatisticalanalysestoidentifythefactorsunderlyingthe5-mercompositionofmobileelementgenomesinthedomainArchaea.Themobileelementfamilies, thegenomeGCcontentsandthephylogeneticpositionof thehostat the taxonomic levelof theorderwere identifiedasmajorexplanatoryfactors. Genomes overall grouped according to the host order, except for thehaloarchaea,allbelongingtotheclassHalobacteria,whichformedasinglesupergroup.Withineachgroup,cellstendedtoclustertogetherwhilevirusesandplasmidstendedtoclusteraccordingtotheirowntaxonomicfamily.Thiswasonlyageneraltrendasanumberofexceptionswereobserved,withaninhomogeneouslevelofshufflingacrossthe dataset. The observed pattern likely results from the combined influence of co-evolution and environmental constraints, highly contrasted among those archaeawhichcomprisemethanogensanddiverseextremophiles.Itconfirmsthepotentialofk-mersignalforextrachromosomalcontiganalysis[1,2,4],in particular for their taxonomic assignation. Another application could be thedetection of singular evolutionary trajectories by focusing on outliers. Indeed,previously-knownandonenewcaseofrecenthosttransferswereefficientlydetectedduring thepresent study. K-mer approaches rely on a distinct informational contentthanthemoreelaborategenesharingnetworksandappearascomplementary.References1, Galiez C et al, Bioinformatics 2017, 33:3113-3114; 2, Iranzo J et al, Journal of Virology 2016,90:1104311055;3,RenJetal,Microbiome2017,5(1):69.4, Krawczyk PS et al,Nucleic Acids Research2018:gkx1321-gkx1321.

ControlandmaintenanceofprophagesinSalmonellaentericaAstridWahl,AliceBoulanger,AuréliaBattesti,NicolasGinet,MireilleAnsaldiLaboratoire de Chimie Bactérienne, UMR7283, Institut de Microbiologie de la Méditerranée, CentreNationaldelaRechercheScientifique,Aix-MarseilleUniversité,Marseille,Franceawahl@imm.cnrs.fr

Thankstotheincreasingamountofgenomicdata,theimportanceofbacteriophagesinhorizontal gene transfer (HGT) is largely acknowledged. Bacterial genomes typicallyharbormultipledefectiveorfunctionalprophagescarryingawiderangeofgenesthatpotentially provide new abilities. On one hand, these genetic elements may beselectedbecausetheyconferaphysiologicaladvantage,suchasnewvirulencefactorsor adaptive traits. On the other hand, uncontrolled expression of prophage genescouldbedetrimental to thehost.Thus,genomeevolutionbyHGTrequiresapreciseequilibriumbetweentherepressionandexpressionofnewlyacquiredgenes.Ourgoal is to identifygeneralstrategies,otherthanphagerepressorbased, thatareresponsible for the maintenance of prophage genes in bacterial genomes. In thiscontext,wehavepreviouslyshownthatthetranscriptionterminatorRhoisinvolvedinprophage maintenance in E. coli (Menouni et al, 2013). Here, we use Salmonellaenterica serovar Typhimurium, a primary enteric pathogen, as a model. Usually, S.entericagenomescarry4-5functionalprophagesandtheirgenesmakeuparound30%ofthepoolofaccessorygenes.ToinvestigatetheeffectofglobalregulatorsonthemaintenanceofS.entericaspecificprophages, we designed a targeted approach: the excision level of functional anddefective prophages was tested in various mutants of these bacterial regulators byusingmultiplex and quantitative PCR.Our results show thatHNS, a known negativeregulator of horizontally acquired genes, negatively controls the excision of Gifsy1prophage.IncontrasttophagelambdaorP22,inductionoftheGifsyprophagefamilyisnotduetorepressorcleavage,butratherto itssequestrationbyanAnti-repressorproteinproducedwhen theLexA-dependantSOS-response isactivated (Lemireetal,2011).WeshowthattheGifsy1-encodedanti-repressorgenegfoAistheactualtargetof HNS and propose the following model: In the absence of HNS, more GfoA anti-repressor protein is produced which then sequesters the GfoR repressor leading toinduction of Gisfy1 prophage.Wewill perform further experiments to clarify if HNSdirectlyregulatesgfoA.Ifthisisthecase,whereexactlydoesHNSbindincomparisontoLexAforwhomawellconservedbindingbox ispresent inthepromoterregionofgfoA?Understandinghowthebacterialhostpreventsprophageinductionwillgiveaclueonthe mechanisms involved in cooptation and co-evolution of prophages within theirhost.

Type: : oralKey-words : HGT,prophages.

Characterizationandmodellingofphagespopulationstructureinsmearwashed-rindcheeseundervariousenvironmentalripeningconditions.EricDUGAT-BONY,Marie-AgnèsPETIT,JulienLOSSOUARNandStéphaneCHAILLOU*.InstitutMICALIS,INRA,AgroPArisTech,UniversitéParis-Saclay.Jouy-en-Josas.GénieMicrobiologiquedesprocédésetdesaliments,INRA,AgroParisTech,UnviversitéParis-saclay,Grignon.

Counting and characterizing phages population and diversity in complex microbialecosytemsisadifficulttask.Moreover,phageshavehighlymosaicgenomesandtheirassembly out of metagenomic sequencing data is still a challenge that is notcompletely solved.Without the reconstruction of full phage genome sequences it isdifficulttoassessthebacterialhosttargetedbythephagespresentintheecosystemsand thus to assess the role of these viruses in the regulation of the bacterialpopulation.

In the frame of the VIROME ACCESS projet (INRA metaprogram on microbialecosystemandmeta-omics),wehavesetupastrategytoimproveourcharacterizationofphagesgenomicdiversityincomplexecosystems(solidmatrix)choosingtherindofsmear-type cheese (Epoisses) as a model. We have first improved our viral DNAextraction from these ecosystems by comparing several purification steps andanalyzing the yield of viral DNA and the quality of phage genome assembly fromshotgunpair-endMiSeqsequencing.Oursecondstrategywastoapplyour improvedprotocol to an experimental design in which ripening conditions were modulatedaccording to twophysicalparameters (levelofoxygenand temperatureof ripening).Using factorial central compositedesign, the structureofbacterial andyeast,whosediversitywere characterizedby ITS and 16S rDNAV3-V4 amplicon sequencing,weremodelledaccording to theseparameters. Inparallel,viralDNAweresequencedandthe different phages genomes were assembled and characterized for life style(virulent,lysogenic)andtheirpotentialbacterialhost.Thepopulationsofphageswerealsomodelledaccordingthephysicalparametersappliedduringtheripeninginorderto correlate the changes in phages populations to those observed in the bacterialpopulations.

The possibility to extrapolate our strategy to more complex ecosystems will bediscussed.

Type:oral

ProphagesandothermobilegeneticelementsasregulatoryswitchesFannyWegner,EduardoRochaGénomiqueévolutivedesMicrobes-InstitutPasteurdeParis,CNRSUMR3525

Activelysogenyisaformofphage-hostinteractioninwhichtemperatephagesareintegratedintofunctionalopenreadingframes.Controlledexcisionunderspecificconditionsleadstothecompletereconstitutionofthegene,whilstre-integrationoftheseelementsleadstothehaltofitsexpression.Thisprovidesacomplexregulatorymechanismofbacterialgeneexpression.Suchregulatoryswitcheshavebeenobservedingenesrelatingtocompetenceorsporulation,andcanoccurwithothermobilizableelementsbeyondphages.

Here,wepresentthescreenofthecompletegenomesof363Salmonellaentericaand437Escherichiacolistrainsforthepresenceofinterruptedgenesandthepotentialofintactorcrypticprophages(aswellasotherintegrativeelements)toactasregulatoryswitches. Our findings indicate that interrupted genes are particularly enriched forfunctions relating to replication, recombination and repair. We further identifycandidatesforexperimentalvalidation.

Active lysogenyhighlightsthepotentialofmicrobestoco-adapttheirmobilegeneticelements towardsgenetic regulation,which iskey forpathogenicityoradaptationtonewenvironments.

Type: : posterMots-Clés : lysogeny;coevolution

PHAGE-HOST MOLECULAR INTERACTIONS

FunctionofGroELduringbacteriophageinfectionofBacillussubtilisLiaMarquesGodinho1,AudreyLabarde1,EricJacquet2,PauloTavares11DépartementdeVirologie,InstitutdeBiologieIntégrativedelaCelluleCNRSUMR91982InstitutdeChimiedesSubstancesNaturellesCNRSUPR2301

Molecular chaperones play an essential role in the folding of nascent chainpolypeptides, refolding of misfolded proteins, and other house-keeping and stress-relatedfunctions.GroELisknowntoplayaroleinviralinfectionmediatingthefoldingofcapsidproteinsduringmorphogenesisofseveralEnterobacteriaceaephages.SPP1isalyticbacteriophageinfectingthebacteriumBacillussubtilis.SPP1isadsDNAvirusthatbelongstotheSiphoviridaefamilyandactsasamodelsystemforvirusesofGram-positive bacteria. During its infection cycle, SPP1 massively hijacks the host'sresources,leadingtosynthesisof>9MbpofviralDNAand150000polypeptidechainstoproduce200virionsin30minutes,highlychallengingthecell.OurgoalinthisworkwastoinvestigatethedependenceofSPP1onB.subtilisGroEL.We constructed an isogenic strain of B. subtilis with a knock-down of the essentialgene groEL taking advantage of an available library of knock-down mutants of B.subtilis by CRISPR-Cas9. The infection dynamics of SPP1, as well as SPP1 DNAreplication and viral particle assemblywere investigated using quantitative PCR andfluorescence microscopy, respectively. Formation of viral particles was the mostaffected step of the SPP1 infection cycle. Electron microscopy of phage structuresfound in lysates followedbywesternblotanalysis revealedadefect inphagecapsidassemblyduringinfectionofthegroELknock-downmutant.Thisknock-downhasalsoamajorimpactintheinfectionofBsubtilisphagesphi29andSPO1.Thestep(s)oftheviralcycleaffectedisunderinvestigation.

Type: : oralMots-Clés : Bacillussubtilis;SPP1;chaperonins.groEL

AsinglephageproteinefficientlydisruptsthehostphysiologybytargetingsigmafactorsMathieuDeJode,AnneChevallereau,LaurentDebarbieuxInstitutPasteurUniversitéSorbonneParisCité(USPC)

Abacterialcellinfectedbyaphagebecomesaviralfactory(orvirocell)asmostofitsresourcesarededicatedtovirionsproduction.Toinvestigatethistransformationofthebacteria into a virocell,we are studying the infection of the opportunistic pathogenPseudomonas aeruginosa by the phage PAK_P3. Using a global transcriptomicapproach, we found that PAK_P3 alters the transcription of over a thousand hostgenes and temporally regulates the expression of its own genes. Amongst the earlyexpressedgenes,wearenowfocusingourattentionongp92,whichisoneofthefewnon-structuralgenesconservedbetweenKpp10virusandPakpunavirusgenera.We discovered that ectopic expression of gp92 in P. aeruginosa alters cellsmorphology:theybecomesphericalinsteadofrodshaped.Thischangeinmorphologydoesnotaffectcellgrowthratenorcellviability.AdditionalmolecularstudiesrevealedthatGp92possessesanunusualmembraneanchoringamino-terminalsequencethatisabsolutely required for this phenotype. Next, using a bacterial two-hybrid assayweidentifiedtheanti-sigmafactorMucAanditstargetthesigmafactorAlgUasputativepartnersofGp92.Severalassaysconfirmedthatexpressionofgp92disruptstheAlgUmediated membrane stress response. Moreover, a mass spectrometry analysisrevealed thatexpressionofgp92 leads toa largemodificationof the cell proteome,includingtheoverexpressionofRpoHandFliA,twoothersigmafactors.Therefore,theexpression of this single phage gene, affecting three sigma factors, alters theregulationofexpressionofalmost1000host'sgenes.WeproposethatearlyexpressedproteinslikeGp92allowphagestoefficientlydisruptthehostcellphysiology,andtransformabacterialcellintoaviralfactory.

Type: : oralMots-Clés : phage;host;protein;sigmafactors

Effets de la chaleur et du chlore sur l'inactivation et les caractéristiques physico-chimiquesdubactériophageMS2AdrienBrié1,2,ChristopheGantzer1,NicolasBoudaud2,IsabelleBertrand11 Laboratoire de Chimie Physique et Microbiologie pour les Matériaux et l’Environnement, CentreNationaldelaRechercheScientifique:UMR7564,UniversitédeLorraine2ACTALIAFoodSafetyDepartment

Lesvirusentériquespathogènesetlesbactériophagessontsoumis,endehorsdeleurhôte,àdenombreuxfacteursconduisantàleurinactivation.Parmieux,lachaleuretlechlore sont les traitements les plus fréquemment utilisés pour inactiver les virusrespectivementdans l'industrieagro-alimentaireet letraitementdeseaux.Leseffetsde la chaleur et du chlore sur les caractéristiques physico-chimiques des particulesvirales sont encoremal connues.Nos travauxont été réalisés avec lebactériophageARN F-spécifique MS2 (1014 UFP/mL) utilisé comme modèle des virus entériquespathogènesetnotammentdesnorovirus.Aucoursdel'expositionà lachaleur(60°Cpendant10,30et60min), l'inactivationaatteint -1à -2,7 log10etunediminutionde la chargeélectrostatiquenégativeaétéobservée.Defaçonintéressante,uneaugmentationdel'hydrophobiedesurfaceaéténotée uniquement pour les phages ayant conservé leur caractère infectieux. Aprèsexpositionàcesconditions, lesphagesont tousconservé leur résistanceà laRNase.Par contre, l'expositionàune températurede72°Cpendant10mina conduit àuneinactivationmassive (-8 log10)avecrupturedescapsidesd'après lesobservationsdemicroscopieélectroniqueàtransmissionconfirméesparlestestsàlaRNase.L'expositiondesphagesMS2à l'hypochloritedesodium(100et200mg/L,10min)aentraînéune inactivationde -1,2et -3,5 log10,etune faiblediminutionde lachargeélectrostatiquenégative;aucunemodificationdel'hydrophobieoudelaperméabilitéde la capside à la RNasen'a été notée. L'utilisationd'une sondehydrophobe SYPROOrange permettant de déterminer le point de rupture des protéines a montré unediminutionduTmdelaprotéinedecapsidesuggérantsafragilisationparlechlore.Lorsque le phageMS2 a été exposé au chlore (100mg/L, 10min) puis à la chaleur(55°Cou60°C,10min),uneinactivation(-1,9et-3,2log10)etuneperméabilitédelacapsideàlaRNasesupérieuresàcellesobtenuespourcesdeuxtraitementsisolésontétéobservées.Lamodificationlaplusmarquanteaétéunetrèsforteaugmentationdel'hydrophobie de surface uniquement pour les phages MS2 ayant conservé leurcaractèreinfectieux.Ainsi,lestraitementsassociantcesdeuxparamètresdevraientutiliserlechlorepuislachaleurpouruneffetoptimal.L'expressiondedomaineshydrophobesàlasurfacedescapsidesaucoursdestraitementsauxquelssontsoumislesviruspourraitjouerunrôleimportantdansladistinctionentrevirusinfectieuxetinactivés.

Type: : oralMots-Clés : MS2;chaleur;chlore;inactivation;hydrophobie

Virulent coliphages isolated from 1 year-old children gut microbiota are sub-dominantcomparedtotemperates,buthighlyinfectiousMarie-Agnès Petit1,2, AurélieMathieu, SylvainMoineau,MoïraDion,Denise Tremblay, LingDeng,DennisNielsen,JakobStokholm,HansBisgaard1:INRAInstitutNationaldelaRechercheAgronomique2:MICrobiologiedelÁLImentationauServicedelaSantéhumaineInstitutNationaldelaRechercheAgronomique:UMR1319,AgroParisTech

Alinkbetweenone-yearoldinfantgutmicrobiotaandasthmapredispositionstartstoemerge from several studies. Since bacteriophages constitute an importantcomponentofthegutmicrobiome,itisofhighinteresttoinvestigateiftheycouldinfluenceearlylifemicrobiotacolonizationandmaturationandwhethertheyplayaroleintheriskoflater asthmatic disease development. To do that, we collaborate within ‘Earlyvir'Europeanjointinitiativeaimingatstudyingtheinfantviromes.Since host prediction from phage sequences still represents a bio-informaticschallenge,culturomicsmightbeaninterestingalternativetoapprehendwhichspeciesofthemicrobiotaarephagetargets.Escherichiacoliisoneofthefirstbacterialspeciestocolonizetheinfant'sgut,andstillrepresents6%oftotalOTUsat1year.In 24% of the virome samples, coliphages could be directly cultivated on indicatorstrains.A collection of 75 purified coliphageswas further studied for its host range, over aselection of 90 E. coli strains representative of the cohort. 28% of them werevirulent,theremainingonestemperate.Strainresponsewasmarkedlydifferentinthetwo phage groups:whereas 93% of the tested strainswere sensitive to at leat onevirulentphage,only18%ofthestrainsweresensitivetoatleastonetemperatephage.The host rangewas also very different in the two groups, with 30% of the virulentphageskillingatleast30%ofthestrains,whereasnoneofthetemperatephagescouldkill30%ofthestrains.Weconcludethatthevirulentfractionhasmostlyallinfectivepower,andbroadhostrange. Inthe lightofviromesequenceanalyses, thissuggeststhatnotallphageswillbeequalintermsofimpactonthemicrobiota.

Type: : oralMots-Clés : phAPEC8;ESSI;2;VpaE1;Stevie;Gluttony;T5

Nitricoxide-drivenprophagemaintenanceinvolvesunsuspectedactivityofNorV(W)reductaseStephanie Champ1, Maëlle Delannoy1, Alice Boulanger2, Yann Denis3, Gaël Brasseur4,MireilleAnsaldi51 Laboratoire de Chimie Bactérienne,UMR7283, Institut deMicrobiologie de laMéditerranée, CentreNationaldelaRechercheScientifique,Aix-MarseilleUniversité,Marseille,FranceCNRS:UMR7283,2Laboratoire des Interactions Plantes-Microrganismes (LIPM) - Institut national de la rechercheagronomique(INRA):UMR2594/44124chemindeBordeRouge-AuzevilleCS5262731326CASTANETTOLOSANCEDEX-France3Institut de Microbiologie de la Méditerranée, Centre National de la Recherche Scientifique, Aix-MarseilleUniversité,Marseille, France (IMM) -CNRS :UMR7283,Aix-MarseilleUniversité–AMU31CheminJosephAiguier,13402Marseillecedex20-France4Laboratoire de Chimie Bactérienne, UMR7283, Institut deMicrobiologie de laMéditerranée, CentreNationaldelaRechercheScientifique,Aix-MarseilleUniversité,Marseille,France5Laboratoire de Chimie Bactérienne, UMR7283, Institut deMicrobiologie de laMéditerranée, CentreNationaldelaRechercheScientifique,Aix-MarseilleUniversité,Marseille,France

Bacterialgenomescontainlargeamountsofprophages,eitherfunctionalordefective.All these prophages contribute to bacterial genome evolution by providing anadditionalpoolofgenesthatsometimesaccommodatenewpropertiestothehost,aphenomenon called lysogenic conversion. Prophagesusually highjack thehost stressresponsesignalisationtoresumealyticcyclewhenconditionsbecomethreateningforthehostandthereforefortheintegratedprophage.This work shows that nitric oxide serves as a maintenance signal and induces theproduction of NorV(W) reductase, which in turn prevents prophage inductionindependently of its usual activity. Surprisingly, nitric oxide, which is a potentnitrosativeagentresponsibleformanycellulardamages,doesnotpromoteprophageinduction but rather maintains and counteracts the SOS-response outcome.Counteracting prophage inductionmakes particularly sense for enterobacteriawhenexposedtonitricoxideproducedinthegutduringinflammationorthroughtheirownanaerobicrespiration.

Type: : posterMots-Clés : Prophages;Maintienlysogénie;détoxificationduNO;NorVW;

RépresseurdelyseCI

AppY,aphage-encodedproteincentraltothebacterialregulatorynetworkNaoualDerdouri1,NicolasGinet2,MireilleAnsaldi2,AuréliaBattesti21Laboratoiredechimiebactérienne,UMR7283,InstitutdeMicrobiologiedelaMéditerranéeAix-MarseilleUniversité-AMU,CNRS:UMR72832Laboratoiredechimiebactérienne,UMR7283,InstitutdeMicrobiologiedelaMéditerranéeAixMarseilleUniversité:UMR7283,CentreNationaldelaRechercheScientifique:UMR7283

Bacterial genome diversity is largely due to prophages, which are viral genomesintegrated into theonesofbacteria.Mostprophagegenesare silent,but those thatare expressed can provide new properties to their host. In particular, AppY, atranscriptionalregulatorencodedbytheDLP12prophageinE.coliK12,increasesthelevel of RpoSwhen overproduced. RpoS is themajor sigma factor during stationaryphase and under many stress conditions in γ-proteobacteria. This factor is highlyregulated,inparticularviachangesinproteinstability.Indeed,onceproduced,RpoSisactively degraded by the ClpXP protease. To be recognized as a protease substrate,RpoS interacts with an adaptor protein called RssB, which brings RpoS to thedegradationmachinery.MyrecentresultsshowthatAppYdirectlyinteractswithRssB,the adaptor protein, suggesting a new anti-adaptor function for this protein,consideredsofarasactingonlyontranscription.Moreover,ithasbeensuggestedthatAppYcould regulatearound30genes in thecell. Inorder to identify thesepotentialAppY targets, we performed RNA-seq experiments. The characterization of some ofthe targets shows that AppY is involved in resistance to acid stress but also in theinhibitionofflagellasynthesisand inbiofilmformation.Overall,ourresultssuggestabroader role for AppY than previously anticipated. This work brings to light theregulatorypathwaysexistingbetweengenesfrombacterialandphageoriginsaswellastheimpactoftheseregulationsonbacterialphysiologyandadaptationtodifferentstressconditions.

Type: : posterMots-Clés : stressresponse;E.coli;regulation;lysogeny

Analysede l'adhésinedesphagesde typeT4et recherched'effet synergiqueentrephageslytiquesetbio-moléculesantibactériennesSabrinaN.Trojet1,2BouchraElKhalfi2,AbdelazizSoukri21LaboratoiredeMicrobiologieetGénétiqueMoléculaire,CNRS,Toulouse,FRANCE2Laboratoire de Physiopathologie Génétique Moléculaire & Biotechnologie, Faculté des sciences AinChock,CentrederechercheSanté&Biotechnologie,UniversitéHassanIIdeCasablanca,MAROC

L'utilisation des bactériophages en thérapie contre les infections bactériennesmultirésistantes aux antibiotiques est une alternative prometteuse. Pour cela, laconnaissance détaillée des interactions moléculaires entre le phage et son hôtebactérien,déterminantlaspécificité,estnécessaire.Lespectred'hôted'unphageestmajoritairement déterminé par les adhésines phagiques. Leur reconnaissancespécifiquedesrécepteurssituésà lasurfacebactérienneconstitue lapremièreétapedel'adsorptiondesphages.ChezlamajoritédesphageslytiquesdeTypeT4,detellesadhésinessontsituéesàl'extrémitédesfibrescaudaleslonguesetcodéesparlegène38.L'adhésinegp38estcomposéedequatresSegmentsHyperVariables(HVS)séparésparcinqMotifsRichesenGlycinetrèsconservés(GRM).Cependant,lesdéterminantsde l'adhésine impliqués dans sa spécificité et dans lemaintien de sa structure sontencoremalconnus.Ainsi, lerôledesdifférentscomposants(HVSetGRM)aétéexaminéparuneanalysecomparativedenombreusesadhésinesgp38, suivied'uneanalyseduspectred'hôte.Une corrélation a été observée entremotif d'HVS et récepteur bactérien reconnus.Enfin,unemodificationdelaspécificitédesadhésinesaétéobtenuegrâceàl'échanged'HVS entre deux phages. Cette analyse apporte de nouveaux éléments surl'interactionentre lephageetsonhôtebactérien,quipourraientêtreexploitéspouraméliorerlathérapieantibactérienne.Aussi, comme décrit dans de précédents travaux, l'utilisation de l'effet synergique(l'effet PAS) entre phage et antibiotique dans un traitement pourrait s'avérer plusefficace. C'est dans cette perspective que les travaux en cours sont dirigés versd'autres alternatives thérapeutiques. En effet, certaines molécules antibactériennescommecelled'huilesessentiellesauxpouvoirsantibactériensetpermettantuneffetsynergiqueaveclesphagessontencoursd'exploitation.Pourfinirnousavonsisolédenouveaux phages lytiques, recueillis sur deux sites naturels au Maroc, ayant unespécificitépourdeuxsouchesbactériennespathogènesdel'Homme(isoléesenmilieuhospitalier). Ces isolats phagiques et bactériens constituent des éléments précieuxpour enrichir notre banque de bactériophages spécifiques des souches pathogènesrencontréssurleterritoireMarocain.Deplus,uneétudecomparativedesinteractionsmoléculaires entre les systèmes hôtes-phages étudiés à Toulouse et ceux isolés auMaroc, suivi d'une détermination de la meilleure molécule provoquant un effetsynergique lors de thérapie combinée, pourront amenées des éléments importantdansl'utilisationdephagesenthérapieoudansd'autresapplications.

Type: :posterKey-words:T4

CharacterizationofthelongelusivenucleaseofbacteriophageT5OmbelineRossier,LéoZangelmi,MadalenaRenouardandPascaleBoulangerVirologyDepartment,InstituteforIntegrativeBiologyoftheCell,CNRSUMR9198,Gif-sur-Yvette

Phage T5 infects Escherichia coli and injects its genome in an original two-stepmechanism:intheFirstStepTransfer(FST),only8%ofthephageDNAentersthehostcell. The transfer pauses then for several minutes before DNA entry resumes tocompletion (SecondStepTransfer,SST).TheFST isaccompaniedbyavery rapidandmassivedestructionofbacterialDNA(50%decrease in labeledDNAwithin4minofinfection).TheidentityofthephageT5DNasehasremainedelusiveforsixtyyears,asnone of the phage proteins encoded on the FST-DNA resemble known nucleases.However,A1,agenecarriedbyFST-DNA,appearstocontrolhostDNAdegradationaswellastheSST.Inthisstudyweinvestigatedtheroleofthe62-kDaproteinA1inDNAdegradationinvitroandinthebacterialcell.IntheC-terminalhalfofA1,weidentifiedseveralmotifsthatareconservedinalargefamilyofmetallophosphatasesincludingtheDNArepairandrecombinationnucleasesMre11/SBcD/gp46. Purified A1 exhibited Mn-dependent DNase activity in vitro onssDNA, but not dsDNA.Using fluorescencemicrocopyofE. coli cells,weobserved arapid decrease in bacterial DNA staining with DAPI upon ectopic expression of A1.Moreover,we frequently saw the formationof a focusof fluorescence, suggestingadramatic reorganization of the bacterial nucleoid. Mutations in putative catalyticamino-acid residues abolished nuclease activity in vitro as well as in vivo. Takentogether,ourresultsindicatethatA1isthelongelusiveearly-encodedDNaseofphageT5. Future work will aim to understand how the DNase activity of A1, which actsexclusivelyonssDNAinvitro,facilitatesthedigestionofgenomicdouble-strandedDNAinvivo.

Type:poster

Study of the structure of T5 phage tail tip complex complex using cryo-electron microscopyRomainLinares1,CharlesArnaud2,GregoryEffantin3,GuySchoehn4,CécileBreyton51Institutdebiologiestructurale(IBS-UMR5075)UniversitéJosephFourier-Grenoble1,Commissariatàl'énergieatomiqueetauxénergiesalternatives:DRF/IBS,CentreNationaldelaRechercheScientifiqueUMR5075,UniversitéGrenobleAlpesInstitutdeBiologieStructurale,71avenuedesmartyrs,CS10090,38044GrenobleCedex9-France2Institutdebiologiestructurale (IBS -UMR5075)Commissariatà l'énergieatomiqueetauxénergiesalternatives : DRF/IBS,Centre National de la Recherche Scientifique : UMR5075,Université GrenobleAlpes InstitutdeBiologie Structurale Jean-Pierre Ebel, CEA-CNRS-UJF, 41, rue JulesHorowitz, F-38027Grenoble,Cedex1-France3InstitutdeBiologieStructurale4:Institut de biologie structurale - Commissariat à l'énergie atomique et aux énergies alternatives :DRF/IBS,CentreNationaldelaRechercheScientifique:UMR5075,UniversitéGrenobleAlpesInstitutdeBiologieStructurale,71avenuedesmartyrs,CS10090,38044GrenobleCedex9-France5:Institutdebiologiestructurale(IBS-UMR5075)-CEA,CNRS:UMR5075,UniversitéJosephFourier-Grenoble Institut de Biologie Structurale, CEA-CNRS-UJF, 71, Avenue des Martyrs CS10090, F-38044Grenoble,Cedex9-France

T5phageisacaudalvirusfromtheSiphoviridaefamily,whichaccountsforabout60%of thetailedphagesandcharacterizedbya long flexible tail.At itsdistalend,T5tailharbours the tail tip complex, which is involved in cell recognition (through thereceptorbindingproteinpb5), cellwallperforationandsafeviralDNAchanneling tothebacteriumcytoplasm.

Weaimatobtainingstructuralinformationaboutthetailtipcomplex,beforeandafteritsinteractionwiththemembranereceptorFhuA,aslittleisknownonthemechanismsleadingtoDNAejectionafterbindingtothehost.Here,wepresentpreliminaryworkaboutthestructureofT5tailtipcomplex,usingmainlycryo-electronmicroscopy(cryo-EM)andsingleparticleanalysis.

WeproducedT5phage tails from theT5amD20mutantphage,whichhas anambermutationinthemajorcaspidproteingene,allowingtheproductionoffullyfunctionaltails. T5 tailswere then prepared for cryo-EM (ie. vitrified) and ~ 3200micrographswere acquired using state of the art Titan Krios electronmicroscope and K2 Gatandirectelectrondetector.10500individualT5tailtipparticleswereobtainedfromthesemicrographs and are currently being processed to obtain a high-resolution 3Dstructure.

Thisisworkinprogressandmoreresultswillbepresentedontheposter.

Type: : poster

Mots-Clés : cryo;electronmicroscopy;membrane/pathogeninteraction;T5phage;tailtipcomplex

THERAPY AND BIOTECHNOLOGY APPLICATIONS

VirusesandmembranevesiclesproducedbydenitrifyingbacteriaRoose-AmsalegC1.,FedalaY2.,Venien-BryanC3.,BoccaraM.2,3

1UMR6553ECOBIO,UniversitédeRennes135042RENNESCedex.2InstitutLangevin,ESPCIParis,PSLResearchUniversity,1rueJussieu,75005Paris,France.3AtelierdeBioinformatique(ABI)ISYEB,UMR7205MNHN,45rueBuffon75005Paris.

Excessinputofnitrogenfromagricultureandwaterfacilitiesareknowntopolluterivers(Xiaetal. 2018). To overcome this problem, denitrifying bacteria are currently used in waterpurification,totransformnitrate intodinitrogen.Howeverdenitrificationisnotperformedatmaximum yield despite favourable physicochemical conditions. We hypothesize that.bacteriophages could influence denitrification in sewage treatment plants or river as it hasbeendemonstratedforlong,inindustrialfermentations(deMeloetal.2018).To test this hypothesiswe studied twodenitrifyingbacteria isolated from river.We showedthepresenceintheirgenomeoftemperateviruses,whichwegeneticallyandmorphologicallycharacterized. Using interferometric microscope, we observed in mitomycin-induced phagelysates, theproductionofa largeamountofparticles lessdenseandmoreheterogeneous insizethanphages.Weinterpretedthemasmembranevesicles.Membranevesiclesinadditionofbeingthevehiclesofproteinandnucleicacidsbetweenbacteriahaverecentlybeenshowntobevehicles forviruses (Tzipilevichetal.,2017).All theseobservationswillbediscussed inlightofefficiencyofdenitrificationprocessinsewagetreatmentplantsorriversediment.Xia X., et al .(2018) The cycle of nitrogen in river systems: sources, transformation, and flux.Environ.Sci.:ProcessesImpacts,20,863–891.Tzipilevich E, HabushaM, Ben-Yehuda S. (2017) Acquisition of Phage Sensitivity by Bacteria throughExchangeofPhageReceptors.Cell.168(1-2):186-199.GonçalvesdeMeloA.,LevesqueS.,MoineauS.(2018)Phagesasfriendsandenemiesinfoodprocessing.CurrentOpinioninBiotechnology49:185-190

Type:poster

AssessingphagetherapyagainsttheplantpestXylellafastidiosa

FernandoClavijo1,MireilleAnsaldi2,Marie-AgnèsJacques31Institutnationaldelarechercheagronomique(INRA):UMR1345,BiolineAgrosciences,équipeR,Aix-MarseilleUniversité–AMU31CheminJosephAiguier,13402Marseillecedex20-France2Laboratoire de Chimie Bactérienne, UMR7283, Institut deMicrobiologie de laMéditerranée, CentreNational de la Recherche Scientifique, Aix-Marseille Université,Marseille, France (CNRS-LCB AMU) - CNRS :UMR7283,Aix-MarseilleUniversité –AMU31Chemin JosephAiguier, 13402Marseille cedex20-France3Institut de recherche en Horticulture et Semences (IRHS) - Institut national de la rechercheagronomique (INRA) : UMR1345,Agrocampus Ouest,Université d'Angers IRHS 42 rue GeorgesMorel49071BeaucouzéCedex-France

Xylella fastidiosa (Xf) is a slow growing Gram-negative plant-pathogenic bacteriumemerging in Asia and Europe. Long-known in the Americas, this xylem-specializedbacterium is associatedwithnumerous socio-economically important plant diseases.Xf is transmitted by xylem feeding insects and belongs to the Xanthomonadaceaefamily. Six different subspecies of Xf have been reported so far: fastidiosa (Xff),multiplex (Xfm),pauca (Xfp),sandyi (Xfs), tashke (Xft)andmorus (Xfmor). Listedasaquarantine pest in Europe, three subspecies (Xfp, Xfm Xff and Xfs) were recentlydetectedinEurope,probablyintroducedthroughcommercialexchangesandcausingavarietyofdiseasessuchastheLeafScorchofOlivetreesinItaly(Jacquesetal.2016;Denancéetal.,2017).IntheabsenceofefficientandauthorizedmethodstocontrolXf,amajorchallengeisto develop environmentally friendly biotechnologies to control this plant disease. Inthis context, we propose the use of bacterial specific viruses, known asbacteriophages, to control Xf infections as recently proposed to control destructivebacterial crop diseases. A few phage cocktails became recently available to treatdiseases caused by Xanthomonadaceae or Ralstonia solanacearum (Buttimer et al.,2017).DifferentaspectsofmyworkdedicatedtophagetherapyagainstXfwillbeaddressed:(i)Usingabioinformaticsapproach,IfirstestimatedtheprophagecontentsofvariousXfgenomesastheymayinterferewithefficientphagesuperinfection,(ii)Iwillreportthe isolationof various lyticphages fromdifferentenvironmental sources suchasXfinfected plants and insect vectors, as well as more generalist sources such as rawsewage influents. To get rid of the difficulties linked toXf culturing, I developed anindirectapproachusingX.albilineansafast-growingorganismandcloserelativeofXfto enrich naturally occurring phages. The characterization of the isolated phages isongoing, andwe are improvingXf culture conditions to assay the host range of theisolatedphagesandtesttheirabilitytoinfectthedifferentsubspeciesofXf.

Type:oral oral

Preliminary studies on pharmacokinetics and pharmacodynamics of virulentbacteriophagestreatingPseudomonasaeruginosaandEscherichiacolilunginfectionsR. Delattre1,2,3, B. Gaborieau2, N. Dufour4, T-T. Nguyen2, J. Guedj2, J-D. Ricard2,5, LaurentDebarbieux1

1:groupeIBBA,unitéBMGE,InstitutPasteur,Paris2:IAME,Inserm,Paris3:serviced’anesthésie-réanimationHôpitalBeaujon,Clichy4:servicederéanimationHôpitalR.Dubos,Pontoise5:servicederéanimationHôpitalL.Mourier,Colombes

While the compassionate treatment of antibiotic resistant infections bybacteriophagesisbecomingmoreandmorefrequentinWesterncountries,dataaboutthe dose and the timing required to ensure a successful treatment remain mostlyempirical.Over the past decade we have isolated several virulent bacteriophages that weretestedinamurinemodelofacutepulmonaryinfection.Hereweinvestigatedinmicepharmacokinetics (concentration-time courses in the body resulting fromadministration of a drug dose) and pharmacodynamics (observed effects resultingfromagivendrugconcentration)ofseveralofthesebacteriophages.First, we record bacteriophage distribution over time into several organs (lungs,spleen, liver, kidneys) and blood compartment after an intranasal or a systemicadministration inhealthyanimals. Second,usingourmodelof acutepneumonia,weobserved over time the bio-distribution in the organs and blood compartment ofbacteriophages administrated by intranasal or systemic routes. In each organ wecomparedtherelativeamountofbacteriaandbacteriophages.Our data provide valuable information on bacteriophages half-life, their eliminationrouteandtheirabilitytoreachthebloodstream.Inthefuture,weaimtoimplementapharmacokinetics/pharmacodynamicsmodelofpulmonaryphage-therapy.

Type:poster

SpanishNetworkofBacteriophagesandTransducterElements(FAGOMA)PilarGarcíaInstitutodeProductosLácteosdeAsturias,IPLA-CSIC.Spain.

Thestudyofbacteriophagebiologyhasdeliveredandstillprovidescluestounderstandmultiplebiosyntheticandgeneregulationprocesses,duetotheirrelativesimplicityincomparison with that of cellular organisms. They have also been a great source ofvaluable biotechnological tools. Because of this, but also due to the environmentalinfluence of phages bymodulating the predominant bacterialmicrobiota alongwiththeir potential applications in infectious therapies and in food safety, research onbacteriophagesmaintainsa constantandsteadygrowth. In this context, theSpanishNetworkofBacteriophagesandTransducerElementscalled“Fagoma”wascreatedin2011tojoinseveralresearchgroupswithanotoriousinternationalprestigethatworkonbasicandappliedaspectsofbacteriophages.Alongthistime,theSpanishNethaspromoted fourmeetingsandseveralexchangesof students. Inaddition, theNethasfacilitatedthecommonanalysisofdataobtainedbythedifferentgroups,thesharingofmethodologiesand the launchof collaborationsaswell as theemergenceofnewideas that have promoted cooperative work between different laboratories, all ofwhichhavecontributedtotheadvanceofknowledge.Ofnote,thecurrentNetisbeinguseful because, at present, the promoting groups have a good knowledge of theinterests and methodologies of the different members of the consortium, thusfacilitatingthesynergiesassociatedtocollaborativeresearch.

Type: : oralMots-Clés : Phagestudies,network,Fagoma

BacteriophagetherapyinWesternmedicineGilbertVerbekenHopitalMilitairedelaReineAstrid,Bruxelles

For hundreds ofmillions of years, bacteriophages – the viral parasites of bacteria –protect Earth’s biosphere against bacterial overgrowth. Today, bacteriophages couldhelp address the antibiotic resistance crisis that affects all of society. The greatesthurdletotheintroductionofbacteriophagetherapyinWesternmedicineisthelackofan appropriate regulatory framework. Belgium is now implementing a pragmaticbacteriophage therapy framework that centers on magisterial bacteriophagepreparations. This keynote-talk will discuss regulatory frames and (magisterial)bacteriophageproductionissues…Inspirationforothers,asimplementedinBelgium…

Type:oral

Characterization of new bacteriophages for the biocontrol of a plant bacterialpathogenLisaLeJoncour,StéphanePoussier,ClaraTorres-BarcelóUniversitédeLaRéunion,UMRPVBMT,SaintPierre,Réunion,France

Bacterialwilt(Bw)causedbytheRalstoniasolanacearumspeciescomplex(RSSC)isamongthemost important plant diseases worldwide, severely affecting a high number of crops andornamentalplants.IntropicalregionssuchastheSouthWestIndianOcean(SWIO),Bwhasamajor negative impact in local agriculture. Genetic studies of the RSSC have resulted in thehierarchicclassificationofthesebacteriaintophylotypes,sequevarsandhaplotypes;andtheirworldwide epidemiology has been extensively studied. The difficulties to control Bw arerelated to high bacterial fitness and genetic diversity, and current intensive agriculturepractices.Theaimsofthepresentstudyweretoisolateandcharacterizenewbacteriophages(phages) capableof infecting theRSSC,whichcouldbeusedasabiocontrol tool.Twenty-sixphages infectingRSSCwere isolatedfromagriculturalsamplescollected inReunion Island, intheSWIO.Hostrangeofphageswastestedmeasuringtheircapacityofreplicationin52RSSCstrains,fromlocalor internationalorigin;and3bacteriafromdifferentbutclosegenera.Wedemonstrate that all phages preferentially attack the most abundant RSSC variant in theregion, sequevar I-31, but harbor a highhost range variability.Wedistinguish specialist andgeneralistphagesregardingtheircapacitytotargetthegeneticdiversityofRSSC,withphagescapableofinfectingphylotypesstillnotdetectedintheIsland.Nonetheless,generalistphagesrestricttheiractiontoRSSCandnoneoftheotherbacterialgeneraaretargeted.Thestudiedphages also show differences in their efficiency at decreasing bacterial growth of themostproblematic strainof theSWIO (sequevar I-31,haplotypeMT035).We find thatefficacyandhost range in these phages are negatively correlated, with specialist phages being moreefficient at controlling the region’s RSSC strain than generalist ones. Importantly, wedemonstratethiskindofevolutionarytrade-offinphagesforthefirsttimetoourknowledge.Allinall,wehavepartiallycharacterizedthefirstcollectionofSWIOphages,andadvancedthebasis for the application of phages as biocontrol measures in the management of Bw. Inparticular, four phages have the ideal host range and efficacy traits to be considered ascandidates for the biocontrol of Bw in the SWIO. We are currently performing molecularanalysisand invivoexperimentsofthe isolatedphages inordertoevaluatetheircapacitytoprotect tomato plants from the bacterial disease. The agro-ecological controlled contextrepresentedbyRSSCaffectingtomatogreenhouses inReunion Islandprovideswithauniqueopportunitytotestphagesinasimplifiedbutnaturalenvironment.

NewOpticalMethodfordetectingandcountingbioticnanoparticles

YasminaFedala1,BoccaraMartine2,ClaudeBoccara31ESPCIParis-PSLResearchUniversity-CNRS2Institutdebiologiedel'écolenormalesupérieure(ENS),Paris,Inserm,CNRS:UMR819745,rued'UlmF-75230Pariscedex05-France3Institut Langevin - Ondes et Images - ESPCI ParisTech, CNRS UMR 7587, INSERM U979 (InstitutLangevin - Ondes et Images)ESPCI ParisTech,Inserm,Université Paris Diderot - Paris 7,CNRS :UMR75871rueJussieu,75238ParisCedex05,France-France

We have developed an optical microscope for detecting, counting and sizingnanoparticles.The principle of the detection is based on the interferences produced between anincident incoherent light and the very weak amount of light scattered by theilluminatedparticle.Moreover,eachdetectedparticleundergoingBrownianmotion is trackedovera fewframesrecordedbythemicroscope.Bycouplingtheinformationdealingwithboththescatteredsignalandthespatialdiffusionof theparticles,onecanreachtheparticlessizesanddensity.In the association with Quattrocento, a structure specialized in the creation ofinnovativecompanies,theLangevinInstituteandPSLhavecreatedtheMyriadestart-upforthedetectionandcharacterizationofnanoparticlesbythisinnovativemethodofopticalinterferometry.Amongthesenanoparticles:viruses,vesicles,metaloxides,etc.,whichopensthewaytoapplicationsinthefieldsofmedicine,biologyandtheenvironment.Theresultsofsometestednanoparticleswillbepresented.

Type: : posterKey-words : Opticalmicroscopie;Interference;nanoparticlesCountingandDetection;

Brownianmotion

INVITROANDINVIVORESISTANCETOTHETHERAPEUTICBACTERIOPHAGE536_P1

GABORIEAUBaptiste,DELATTRERaphaëlle,DEBARBIEUXLaurent,RICARDJean-Damien

Antimicrobial resistance is a major public health problem, especially in patientssubjectedtoastrongantibioticselectivepressuresuchasthosehospitalizedinintensivecare units and developing ventilator-associated pneumonia. Enterobacteria, includingEscherichia coli, are a critical priority target for research and development oftherapeutic alternatives. Phage therapy is promising but emergence of bacteriabecomingresistanttobacteriophagesduringtreatmentsisunderstudied.WeisolatedandcomparedbacterialclonesoftheE.colistrain536thatwereselectedas resistant to Myoviridae virulent bacteriophage 536_P1 from both in vitro co-evolution experiments and in vivo treatments in a mousemodel of pneumonia. Theresistancerate,thegeneticmutationsidentifiedbywholegenomesequencingandtheimpactoftheseonthefitnessandvirulenceofthestrainswerestudied.Thelevelsofresistantclonesobtainedwere23%invitroand13%invivo.Interestingly,theresistantclonesisolatedinvitroandinvivoatearlytimepointdisplaymutationsinthe samegenes involved in theLPSbiosyntheticpathway.Thesemutations lower thefitness of these clones. At late time point in vivo the resistant clones carry othermutationsingenesinvolvedintheoutercapsulebiosynthesis.Therefore,thepanelofmutationsofbacteriophage-resistantclonesobtainedinvivo iswider than those observed in vitro. This may be the result of the combination ofmultiple mechanisms, probably related to the selective role played by the hostenvironment, including the immune system, on the fitness-cost of bacteriophage-resistant clones. This initial study requires deeper investigations to assess in vivomechanisms of resistance that could account for the failure of phage therapytreatments.

Type:poster

Keywords:E.coli;PhagetherapyPneumonia;Resistance

IsolementetcaractérisationdephagesciblantFlavobacteriumpsychrophilum,bactériepathogènedestruitesd'élevage

ThomasGuiraud1,CécilePhilippe2, Fety Jaomanjaka2, EtienneGontier3, Claire LeMarrec2,MichelLeHénaff11Bordeaux Sciences Agro (Gradignan) Ecole Nationale Supérieure des Sciences Agronomiques deBordeaux-Aquitaine2InstitutdesSciencesdelaVigneetduVinVillenaved’OrnonUniversitédeBordeaux3Bordeaux Imaging Center, Institut National de la Santé et de la RechercheMédicale : US4,InstitutFrançoisMagendie,UniversitédeBordeaux,CentreNationaldelaRechercheScientifique:UMS3420

Les pratiques d'élevage intensif exposent la filière truiticole française à divers défissanitairesparmilesquelsleplusproblématiqueàcejourestl'émergencedelabactérieFlavobacteriumpsychrophilum, l'agentcausalde lamaladiedusyndromedesalevinsde truite arc-en-ciel (Rainbow Trout Fry Syndrom, RTFS). En parallèle aux mesuresprophylactiques visant à limiter la propagation de l'agent pathogène, l'utilisationintensive de traitements curatifs antibiotiques entraîne l'émergence de souchesantibiorésistantesdevenantpréoccupanteà la foispour la filièrepiscicoleetpour lasanté humaine. La phagothérapie est une approche alternative prometteuse àl'antibiothérapie.Danscecontexte,nousavons initiéunecampagnedeprélèvementd'échantillonspiscicoles(eau;organesdepoissonsinfectés)afind'isolerdesparticulesviraleslytiquespourF.psychrophilum.

L'analysedeséchantillonsbiologiquesapermisl'isolementde20souchesvirales.Troissoucheslytiquesontétéretenuespourleurplusgrandpotentieldevirulenceinvitro.LacaractérisationmorphologiqueparmicroscopieélectroniquedecesbactériophagesindiqueleurappartenanceàlafamilledesSiphoviridae.Leséquençagecompletdeleurgénome révèle (i) la proximité de deux flavophages avec un phage précédemmentidentifié (le phage 6H; Castillo et al., 2014) et (ii) l'isolement d'un 3e flavophageoriginal. En parallèle, nous avons testé la capacité lytique en coculture de ces troissouches, seules ou en combinaisons. Les résultats obtenus ont permis de définir, invitro, une formulation du cocktail phagique la plus efficace pour limiter ledéveloppementdeF.psychrophilum.

Cespremiers résultatsprometteurs incitentàpoursuivre l'explorationde ladiversitévirale régionale pour enrichir notre phagothèque afin (i) d'explorer la diversitégénétiquedesflavophagesetdansuneperspectivededéveloppement,(ii)d'optimiserl'efficacitélytiquedescocktailsphagiques.

Type: : poster

Key-words :

bacteriophages;flavophages;flavobacteriumpsychrophilum;RainbowTroutFrySyndrom;Siphoviridae;phagotherapy;aquaculture;truit;Aquitaine

Addressing knowledge gaps in the taxonomy of phages of Lactic acid bacteria:phage-host interactions in the underrepresented generaOenococcus,Weisella andnon-dairyLeuconostocspp

ClaireLEMARREC1,OlivierCLAISSE1,2,FetyJAOMANJAKA1,CécilePHILIPPE1,AmelCHAIB1,CoralieCOUDRAY-MEUNIER3,4,SarahCHUZEVILLE5,ValérieGABRIEL4andCathyFAUCHER41UniversitéBordeaux,BordeauxINP,ISVV,EA4577,F-33882Villenaved'Ornon,France;2INRA,ISVV,USC1366OENOLOGIE,F-33882Villenaved'Ornon,France;

3ISARALyon;4UniversitédeToulouse,LBAEEA4565(LaboratoiredeBiotechnologiesAgroalimentaireetEnvironnementale),UPS,IUT"A«,24rued’Embaquès,F-32000Auch,France;

5Actalia,LaRoche-Sur-Foron

Phages infecting lactic acid bacteria have been the focus of significant researchattention over the past three decades. Phage infection of dairy starter culturesremainsthemaincauseoffermentationfailures inthedairy industry.Owingtotheireconomical importance,dairyphagesbecamethemostthoroughlysequencedphagegroupinthedatabases.Incontrast,ourknowledgeofphagediversityinnon-dairyfoodfermentations is still in its infancy. This is true for the “Leuconostocaceae" family,including the members of the Leuconostoc, Oenococcus, and Weissella genera.Oenococcus oeni drives the second fermentation of wine, called malolacticfermentation (MLF), which reduces acidity, stabilizes wines and protects it fromspoilage.Weissella cibaria,W. confusa, Leuconostoc mesenteroides and L. citreumhavebeen recentlyassociatedwithdifferentFrench sourdoughbread,andalsowithdifferentvegetablefermentations.In this study,we have collected lytic and temperate phages from food samples andfrom indigenous strains isolated from sourdough breads and other vegetablefermentations. DNAs have been extracted from 250 phages/prophages. The geneticdiversity of phages belonging to different taxonomic groups was further explored.Insights intophage-host interactionsprovideessential data for the currentor futureselection, biomass production and inoculation of commercial starters to be used infermentationsandotherbiotechnologicalapplications.

Type:poster

SurveyonprophagesinfectinglacticacidbacteriaWeissellacibariaandW.confusaCoralie Coudray-Meunier1, Valérie Gabriel1, Claire Le Marrec2 and Catherine Fontagné-Faucher11UniversitédeToulouse,UPS,IUT"A",LBAEEA4565;F-32000Auch,France2UniversitédeBordeaux,INP,ISVV,EA4577,UnitéderechercheOenologie,F-33882Villenaved’Ornon,France

Lactic Acid Bacteria (LAB) infection by bacteriophages could be responsible forimpaired food fermentations. However, there are few studies concerning phages infermented products other than dairy foodstuffs.Weissella cibaria andW. confusabelonging to Leuconostocaceae family (also including Leuconostoc, Oenococcus andFructobacillus genera) have been isolated from a wide range of habitats, and areknowntobeinvolvedinseveraltraditionalfoodfermentationssuchasstarchy,cereal-based,meatandfishproducts(Fuscoetal.2015).Moreover,strainsofthesespeciesarerecognized toproduce copious amountsof oligosaccharides andexopolysaccharides,withspecialinterestastexturizingagentsandprebiotics(Bounaixetal.2010).NophagehasbeenyetdescribedtoinfectW.confusaandonlytwophagesinfectingW. cibaria (φYS61 from kimchi and φ22 from Nham) were currently described, bothbelonging to the familyofPodoviridae (Kleppenetal., 2012;Pringsulakaetal., 2011).Besides,severalphagesinfectingW.cibariawereisolatedfromcommercialcucumberfermentation(Luetal.,2012).The aim of the present study was to investigate the presence of prophages inW.cibariaandW.confusa strainspreviously isolated fromFrenchsourdoughs (Robert etal., 2009) and from other diverse biotopes. Prophage excision was achieved withmitomycin C induction and led to observe bacterial lysis for all the strains.Unfortunately,nosensitivebacteriacouldbedetectedforanyoftheputativephagesexcisedfromthestudiedstrains.EvidenceofphageDNAwasthusachievedbyindirectapproach i.e. purification of phage DNA frommitomycin-induced bacterial cultures.Further DNA characterizationwas achieved by conventional restriction analysis, andrepetitive element (Rep)-PCR profile using (GTG)5 primer as a new phage DNAcomparativetechnique.Overall, the resultsof this studyclearlydemonstrate theprevalenceanddiversityofprophagesinthegenomeofW.cibariaandW.confusaLABstrains.

Type:poster

EnologicalenvironmentasasourceofanovelTectivirusCécilePHILIPPE1,MartKRUPOVIC2,FetyJAOMANJAKA1,OlivierCLAISSE1,3,MelinaPETREL4

andClaireLEMARREC1

1Univ.Bordeaux,BordeauxINP,InstitutdesSciencesdelaVigneetduVin(ISVV),EA4577,UnitéderechercheOENOLOGIE,F-33882Villenaved'Ornon,France 2 DepartmentofMicrobiology,InstitutPasteur,Paris,France.3INRA,ISVV,USC1366OENOLOGIE,F-33882Villenaved'Ornon,France4Univ.Bordeaux,BordeauxImagingCenter,UMS3420CNRS-US4INSERM,Bordeaux,France

Aceticacidbacteria(AAB)areubiquitous,strictlyaerobicbacteriaoccurringinsugary,alcoholicandacidicniches.Membersofthegrouphavebeenreportedinarangeoffoodandbeverageecosystems, and play a positive role during the fermentation of vinegar, cocoa bean, kefir,kombucha, and acidic beers (1). In contrast, AAB represent spoilagemicroorganisms duringwinemaking,mainlybecausetheyareabletoproduceethylalcoholandtransformitintoaceticacid. Thegenera AcetobacterandGluconobacterare themost important producers ofwinespoilagewhichislegallydefinedbyvolatileacidity,largelycomposedofaceticacid.DetectionofAAB is reportedatall stages, fromthematuregrape throughvinification toconservation.AABoriginate fromthe leavesof thegrapevineandthegrapesthemselvesandfruit fliesareconsideredasacommonvectorinpropagatingAAB.Differentauthorshaveobservedthatthelesshealthythegrapes,thehighertheamountofAABis.PhagesinfectingAABhavereceivedrelatively littleattention.ThisstudyreportsasurveyofphagesofAABcollectedfromgrapeswhich resulted in the isolation and sequencing of a novel phage infecting G. cerinus,representinganovelmemberofthefamilyTectiviridae.

Type:poster

Participants

AnsaldiMireille LaboratoiredeChimieBactérienne,CNRSAMU ansaldi@imm.cnrs.frAubritFrançoise CleanCells,Bouféré faubrit@clean-cells.frAzeredoJoana U.Braga,Minho,Portugal jazeredo@deb.uminho.ptBattestiAurelia LaboratoiredeChimieBactérienne,CNRSAMU abattesti@imm.cnrs.frBerngruberThomas EVONIKNutrition&CareGmbH,Essen thomas.berngruber@evonik.comBertrandIsabelle LCPMEUMR7564 isabelle.bertrand@univ-lorraine.frBillaudMaud INRAMicalis BizeAriane IRSTEA,Antony ariane.bize@irstea.frBoccaraMartine ENSCRNSUMR8197InsermU1024 boccara.martine@gmail.comBoulangerPascale I2BCCNRSGif-Sur-Yvette pascale.boulanger@i2bc.paris-saclay.frBoyerMickaël nd mickael.boyer38@gmail.comBreytonCécile UBSCNRS,GRENOBLE Cecile.Breyton@ibs.frBrivesCharlotte CNRS,CentreEmileDurkheim charlottebrives@gmail.comChaibAmel ISVVBordeaux,U.Bordeaux-BordeauxINP bounoise68@hotmail.frChaillouStéphane MICALIS.UMR1319INRAAgroParisTech stephane.chaillou@inra.frChatainMaiHuong Vetophage,Lyon chatain@vetophage.frClaisseOlivier ISVVBordeaux,INRA,U.Bordeaux-BordeauxINPolivier.claisse@inra.frClavijoFernando LaboratoiredeChimieBactérienne,CNRSAMU fclavijo@imm.cnrs.fr CostechareyreDenis Bactolytix ddcostechareyre@yahoo.frDauchyFrédéric-Antoine CHUBordeaux frederic.dauchy@chu-bordeaux.frDebarbieuxLaurent InstitutPasteur,Paris laurent.debarbieux@pasteur.frDeJodeMathieu InstitutPasteur,Paris dejodemathieu@gmail.comDelattreRaphaëlle InstitutPasteur,Paris raphaelle.delattre@gmail.comDemarreGaëlle PherecydesPharma,Romainville gaelle.demarre@pherecydes-pharma.com

DePaepeMarianne MICALIS,INRA,Jouy-En-Josas marianne.depaepe@inra.frD'humièresCamille InstitutPasteur,Paris camille.dhumieres@pasteur.frFedalaYasmina ESPCI,Paris yasmina.fedala@espci.frForterrePatrick InstitutPasteur patrick.forterre@pasteur.frFroissartRémy UMR5290MIVEGECCNRSMontpellier remy.froissart@cnrs.frGaborieauBaptiste InstitutPasteur bgaborieau@hotmail.fr GandonSylvain UMR5290MIVEGECCNRSMontpellier sylvain.gandon@cefe.cnrs.frGarciaPilar DairyInstituteofAsturias,Madrid,Spain pgarcia@ipla.csic.esGinetNicolas LaboratoiredeChimieBactérienne,CNRSAMU nginet@imm.cnrs.fr GuiraudThomas BordeauxSciencesAgro thomas@guiraud.co HardyAel ForschungszentrumJülichGmbH aelhardy@hotmail.frJaomanjakaFety ISVVBordeauxU.Bordeaux-BordeauxINP jaomazava@yahoo.frJaramilloAlfonso U.Warwick alfonso.jaramillo@gmail.comKleinElodie IBMP-CNRS,U.Strasbourg elodie.klein@ibmp-cnrs.unistra.frLamy-BesnierQuentin InstitutPasteur quentin.lamybesnier@yahoo.frLeBourgeoisPascal BLADELISBP-INSAToulouse pascal.lebourgeois@univ-tlse3.frLecointeFrançois INRAMICALIS francois.lecointe@inra.fr LeMarrecClaire ISVVBordeaux,U.Bordeaux-BordeauxINP clehenaff@enscbp.frLinaresRomain UBSCNRS,GRENOBLE romain.linares@ibs.fr LossouarnJulien INRAMicalis julien.lossouarn@inra.fr MahonyJennyfer UCC,Irlande J.Mahony@ucc.ieMarcouxPierre LETI–DTBS,CEAGRENOBLE pierre.marcoux@cea.fr MarquesGodinhoLia I2BCUMR9198,GIF-SUR-YVETTE lia.marques-godinho@i2bc.paris-saclay.frMccallinShawna CHUvaudois,EPALINGES shawna.mccallin@gmail.comMedinaMathieu PherecydesPharma mathieu.medina@pherecydes-pharma.com

PerlemoinePrisca LETI–DTBS,CEAGRENOBLE Prisca.PERLEMOINE@cea.frPetitMarie-Agnès INRAMicalis marie-agnes.petit@inra.frPetitjeanCharlotte PherecydesPharma charlotte.petitjean@pherecydes-pharma.com

QuilletLaurent UniversitédeRouen laurent.quillet@univ-rouen.fr RenardAdélaïde U.Tours adelaide.renard@etu.univ-tours.frRocheStéphane I2BCUMR9198,GIF-SUR-YVETTE stephane.roche@i2bc.paris-saclay.fr

RossierOmbeline I2BCUMR9198,GIF-SUR-YVETTE ombeline.rossier@u-psud.frSacherJessica UniversityofAlberta sacher@ualberta.ca SamotJohan ISVVBordeaux johan.samot@gmail.com SchiavolinLionel IBENSParis schiavol@biologie.ens.frSCWEWYWABBITJan PhageDirectory,ATLANTA janeazy@gmail.comTavaresPaulo I2BC,CNRS,UniversitéParis-Sud,CEA tavares@vms.cnrs-gif.frTorres-BarcelóClara UniversitédelaRéunion clara.torres@cirad.fr TrojetSabrina UniversitéHassanIIdeCasablanca sabrina.trojet@gmail.com TrotereauAngelina INRANouzilly angelina.trotereau@inra.frVerbeckenGilbert HopitaldesArmées,Bruxelles Gilbert.Verbeken@mil.beVincentClaude APRC,InterAcademicGroupIAGmicrobiome claude.vincent@l-n-c.frWahlAstrid LaboratoiredeChimieBactérienne,CNRSAMU awahl@imm.cnrs.fr WegnerFanny InstitutPasteur fanny.wegner@pasteur.fr

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