POSTER ABSTRACT BOOK

@MicrobioSoc #EZAMR18

microbiologysociety.org/EZAMR18

2 July 2018

University of Surrey, UK

Emerging Zoonoses and AMR: A Global Threat

FOCUSED MEETING

2018

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AC A4.indd 1 20/03/2018 10:10

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PosterBookContentPosterNumber PresentingAuthor AbstractTitle Page

Number

01MariaGetino

(UniversityofSurrey,UK)AnovelapproachtostudytransmissionofAMRplasmidsincomplexmicrobialcommunities

03

02HelenBrown

(UniversityofSurrey,UK)ManukahoneyinhibitstheviabilityandvirulenceofStaphylococcuspseudintermediusinvitro

04

03JorgeGutierrez-Merino(UniversityofSurrey,UK)

Probioticsasanalternativediseasemitigationinwildlife

05

04RebeccaMcLean(ThePirbrightInstitute,UK)

DevelopmentofaNipahvirusvaccinetoeliminateporcinereservoirsandsafeguardhumanhealth 06

05

PiyaliBasu(UniversityofSurrey,UK)

Usingamulti-disciplinary,onehealth,approachtounderstandinghowemerginginfectiousdiseasesandantimicrobialresistance(AMR)cantransferfromlivestocktohumans

07

06

IsabellaCenteleghe(CardiffUniversity,UK)

Combatingtherisingglobalthreatofantimicrobialresistancewithclayminerals:Astudyontwomajorhospitalsuperbugs.

08

07Mary-AnneFrank(UniversityofPretoria,SouthAfrica)

AsurveyofantibioticresistanceinentericEscherichiacoliisolatedfromungulatesatazoologicalpark 09

08

ElaineMeade(CellularHealthandToxicologyResearchGroup,InstituteofTechnology,Ireland)

ZoonoticantimicrobialresistantCandidaandbacterialspeciesisolatedfromcompanionanimals.

10

09LucyRhys-Davies(UniversityofSurrey,UK)

DevelopingphagetherapyforthetreatmentofE.coliinfectionsincompanionanimals

11

10

SarahJohns(UniversityofSurrey,UK)

AvianPathogenicandcanine,andhumanuropathogenicEscherichiacoli;Reservoirsofantimicrobialresistance

12

11

MaríadelMarFernándezdeMarco(AnimalandPlantHealthAgency,Addlestone,UK)

CompleteUsutuvirusgenomesequencesderiveddirectlyfromclinicalsamplesbyusingamultiplexPCR-basedwholegenomesequencingmethod. 13

2

PosterNumber PresentingAuthor AbstractTitle Page

Number

12ArnoudHMvanVliet(UniversityofSurrey,UK)

Identificationoflineage-specificgeneticmarkersofListeriamonocytogenesbypangenomeanalysis

14

13

AmandaSFivian-Hughes(UniversityofSurrey,UK)

TheimpactofhostrestrictionofEscherichiacoliontransmissiondynamicsandspreadofantimicrobialresistance(HECTOR)

15

14

ConorLarkin(WestwayHealth,Ireland)

IodideCompositionsfortheTreatmentofBovineMastitis 16

15

StevenKemp(UniversityofLiverpool,UK)

AntibioticResistancePatternsandGeneInterplayBetweenAnimals,HumansandtheEnvironmentinaHigh-DensityLivestock-HumanPopulationinWesternKenya

17

16AdamMoore(CardiffUniversity,UK)

Anthraxandclayminerals:anovelsolution.18

3

PosterNumber:01AnovelapproachtostudytransmissionofAMRplasmidsincomplexmicrobialcommunities

Abstract

Thestudyofantimicrobialresistance(AMR)transmittedbyextrachromosomalgeneticelementsiscurrentlylimitedbytheabsenceofapproachesabletolinkhost-specificandplasmid-specificsequenceswithinmetagenomicdatasets.Toovercomethisissue,wehavedevelopedanovelmethodologythatwillallowthetrackingofspecificplasmidsincomplexmicrobialcommunities.Representativebroad-host-rangeandnarrow-host-rangeAMRplasmidsweregeneticallymodifiedtocarryararebacterialmethyltransferase.Whenexpressedinthehostbacterium,thisenzymemethylatesspecificsitesacrosstheentiregenome,includinginspecies-specificgenesusefulforidentifyingorganisms.Single-MoleculeReal-Timesequencingtechnologycanthendetectthegenomicmethylationpatternbasedonpolymerasekinetics.Thusfar,wehaveverifiedmethyltransferaseactivityindifferentbacterialhostscarryingthemodifiedplasmidsbyusingquantitativePCRtomeasuretheextentofDNAprotectionaftertreatmentwiththecognateendonuclease,arestrictionenzymethatdigestsunmethylatedsites.Conjugativetransfer,stability,andfitnessofmodifiedAMRplasmidswasconfirmedtobeunaffectedbythepresenceofthemethyltransferasegene.Asaproofofconcept,invivoassaysusingC57BL/6miceinfectedwithCitrobacterrodentiumcarryingtheengineeredplasmidsareon-going.MetagenomicDNAisolatedfromfaeceswillbecollectedandsequencedtoanalysethespreadoftheseplasmidsintheintestinalcommunity.Thisapproachwillbeusefultoidentifynovelreservoirsandroutesofplasmiddisseminationindifferentenvironments,andtesttheeffectoftreatmentstopreventAMRtransmission.

MariaGetino1,JuhyunKim1,GangFang2,JohnKenny3,JoseJimenez1,JennyMRitchie1

1UniversityofSurrey,Guildford,UnitedKingdom.2IcahnSchoolofMedicineatMountSinai,NewYork,USA.3UniversityofLiverpool,Liverpool,UnitedKingdom

4

PosterNumber:02ManukahoneyinhibitstheviabilityandvirulenceofStaphylococcuspseudintermediusinvitro

Abstract

Staphylococcuspseudintermediusisacommonveterinarypathogenwithsignificantzoonoticpotential.Therecentemergenceofmultidrug-resistant(MDR)S.pseudintermediusshowsthatnoveltherapeuticsolutionsareurgentlyrequired.ManukahoneyinhibitsmanybacterialpathogensincludingmethicillinresistantStaphylococcusaureus,andisusedinbothclinicalandveterinarypractice.Ithasalsobeendemonstratedtoactsynergisticallywithantibioticsinvitro,increasingtheirpotencyandmakingitapromisingadditiontotreatmentregimenswhereMDRispredicted.

HerewedeterminedtheabilityofmanukahoneytoinfluenceS.pseudintermediusgrowth,biofilmformationandpathogenicitybothaloneandincombinationwithclinicallyrelevantantibiotics.Lowconcentrations,≥10%(w/v),ofmanukahoneywereabletoinhibitgrowthof20S.pseudintermediusisolates.Susceptibilitytoselectedantibiotics(gentamicin,chloramphenicol,clindamycin,penicillinG,tetracycline)wassignificantlyincreased(p=<0.05)whencombinedwithsublethalconcentrationsofhoney.Biofilmbiomasswasreducedbytreatmentwithmanukahoney,withlive/deadstainingandscanningelectronmicroscopyshowinginactivationofthecellswithinthebiofilmandalterationsinmorphology.Finally,phenotypicexpressionofvirulencewassignificantlyreduced(p=0.05)inthemajorityofisolateswhentreatedwithsub-lethalhoneyconcentrations.

Thisstudyhighlightsthepotentialformanukahoneytobeutilisedinaveterinarysetting,increasingthesusceptibilityofbacterialpathogenstoantibiotics.Withfurtherinvivotestingthismayofferanalternatetherapytothoseanimalswithinfectionsthatarenotrespondingtoconventionaltherapy,orwhenS.pseudintermediuscausesopportunisticinfectionsinhumans.

GeorgieMetters1,JennaCooper2,LuisSousa3,HarryDance4,RobertAtterbury4,HelenBrown5,RowenaJenkins3

1ExeterUniversity,Exeter,UnitedKingdom.2CardiffMetropolitanUniversity,Cardiff,UnitedKingdom.3SwanseaUniversity,Swansea,UnitedKingdom.4NottinghamUniversity,Nottingham,UnitedKingdom.5UniversityofSurrey,Guildford,UnitedKingdom

5

PosterNumber:03Probioticsasanalternativediseasemitigationinwildlife

Abstract

Bovinetuberculosis(bTB)isachronicbacterialdiseasecausedbyMycobacteriumbovisthatleadstosignificanteconomiclossesworldwide.bTBhasbeeneradicatedfrommanyEuropeannationsbutitisstillveryprevalentinsomecountrieswherewildlifereservoirsofM.bovishavebeenconfirmed.ThisisthecaseofwildboarandtheEuropeanbadger.Lacticacidbacteria(LAB)havebeenproposedasanewalternativeforcontrollingbTBduetotheirprobioticproperties,whichincludetheirabilityto:(1)inhibitthegrowthofMycobacteriumspecies;and(2)triggerbeneficialhostimmuneresponses.ThemainobjectiveofthisstudywastoevaluatethepotentialasofLABisolatedfromfaecesofwildboarandbadgersasaprobioticalternativeagainstbTB.LABhavebeenisolatedandidentifiedasPediococcus,Lactobacillus,EnterococcusandWeissella.Overall,theisolateshaveshownsignificantantimycobacterialactivityandseemtobeassociatedwithinnateimmunomodulation.Someoftheisolateshaveinducedproinflammatorypathways,suggestingapotentialroleasvaccineadjuvants;whereasotherisolateshaveshowedpotentialanti-inflammatoryproperties.Furtherstudiessuchaswhole-genomesequencingandantibioticresistancetestshaveconfirmedthepotentialuseofourLABisolatesasprobiotics.OurdatasuggestthatLABcouldbeusedasvaccineadjuvantstoreduceorpreventinfectionbutalsoasatooltoreduceinflammationandtheamountofviableexcretedmycobacteriainhighlyinfectedanimals.Thesemeasurescouldindeedleadtoalong-termdecreaseintheprevalenceofbTBandotherinfectiousdiseasesinwildboarandbadgers

JorgeGutierrez-Merino

UniversityofSurrey,Guildford,UnitedKingdom

6

PosterNumber:04DevelopmentofaNipahvirusvaccinetoeliminateporcinereservoirsandsafeguardhumanhealth

Abstract

Nipahvirus(NiV)posesasignificantepidemicthreatduetoitsbroadhostrangeandthewidespreaddistributionoffruitbatswhichprovideanaturalreservoir.Humaninfectioncanoccurfollowingexposuretoinfectedpigsresultinginsevereandoftenfatalrespiratoryandneurologicaldisease.Pig-to-humantransmissionwasresponsibleforthefirstandmostsevereNiVoutbreaksinMalaysiaandSingapore.Theseoutbreaksresultedinsignificanteconomiccostsandlong-termdamagetolocalpigindustries.DespitetheimportanceofNiVasanemergingpathogen,novaccinesarecurrentlyapprovedforhumanorlivestockuse.Weproposethataninexpensive,safeandefficaciousvaccinecouldbedevelopedtoprotectpigsagainstNiVinfectionandtransmission,thereforereducingtherisktopublichealthaswellaspigindustriesintheendemicregion.Inthisproject,weareassessingtheimmunogenicityandefficacyofthreeNiVvaccinecandidates:(1)arecombinantNiVGproteinsubunitvaccine;(2)arecombinantNiVFproteinsubunitvaccineand(3)areplicationdeficientadenoviralvectorexpressingNiVGprotein.ThemosteffectivecandidatewillbefurtherassessedtodetermineitsabilitytopreventNiVtransmission,thedurabilityofimmunityandoptimalimmunisationregimen,followedbyevaluationofsafetyandimmunogenicityunderfieldconditionsintwohigh-riskcountries.InadditiontodevelopingaporcineNiVvaccine,datageneratedthroughthisstudywilldirectlyinformeffortstodevelopahumanNiVvaccine.

RebeccaMcLean1,DalanBailey1,NagendraNathBarman2,Li-YenChang3,KeithChappell4,SarahGilbert5,TeresaLambe5,GlennMarsh6,MiriamPedrera1,RudigerRaue7,ElmaTchilian1,NaziaThakur1,OoiPeckToung8,DanielWatterson4,PaulYoung4,SimonGraham1

1ThePirbrightInstitute,Pirbright,UnitedKingdom.2AssamAgriculturalUniversity,Assam,India.3UniversityofMalaya,KualaLumpur,Malaysia.4UniversityofQueensland,Brisbane,Australia.5JennerInstitute,Oxford,UnitedKingdom.6CSIROHealthandBiosecurity,Geelong,Australia.7Zoetis,Zaventem,Belgium.8UniversitiPutraMalaysia,Selangor,Malaysia

7

PosterNumber:05Usingamulti-disciplinary,onehealth,approachtounderstandinghowemerginginfectiousdiseasesandantimicrobialresistance(AMR)cantransferfromlivestocktohumans

Abstract

TheEuropeanJointProgramme(EJP),isaco-fundactiondesignedtosupportcoordinatednationalresearchandinnovationprogrammes.TheEJPaimstopoolacriticalmassofnationalresourcestoworkontheobjectivesandchallengesofHorizon2020.TheUniversityofSurrey,alongwiththeirpartnersacrossEurope,weresuccessfulinobtaining€90millioninordertoinvestigatetheemergenceandtransmissionofinfectiousdiseases,andAntimicrobialResistance(AMR)throughoutthefoodchain.Herewewillpresentanoverviewoftheprojectanditsscope,highlightingthebenefitsoftheonehealth,multidisciplinaryapproachapplied.

TheUniversityofSurreywillfocusprimarilyontwocomplementaryresearchareas.Themicrobiomeofkeyanimalspecies(pigsandpoultry)willbeprobedinordertodetermineifspecificmicrobiotacompositionsarelinkedtothepathogensheddingstatusofthehost.Probioticspeciesandnutraceuticalswillthenbeidentifiedandassessedfortheirabilitytolimitshedding,andsoreducespreadofbacterialpathogenswithinherdsandflocks.Inparallel,themaindriversandagentsinvolvedinAMRtransmissionanddisseminationacrossEuropewillbeinterrogated.Aparticularfocusofthestudieswillbeonmobilegeneticelementsandtheirtransferpotentialbetweenhosts.ItisanticipatedthattheoutputsofthestudieswillresultinreducedzoonoticpathogensinthefoodchainandreducedantimicrobialusageandAMR.Emphasiswillalsobeplacedontrainingofscientists,inordertoensurethesustainabilityoftheresearchareaandcollaborations.

PiyaliBasu,HelenBrown,MariaGetino,DanHorton,ElizabethRoyall,RobertoLaRagione

UniversityofSurrey,Guildford,UnitedKingdom

8

PosterNumber:06Combatingtherisingglobalthreatofantimicrobialresistancewithclayminerals:Astudyontwomajorhospitalsuperbugs.

Abstract

Theglobalincreaseinantimicrobialresistanceisplacingincreasingpressureonhealthcaresystemstoday.Historically,claymineralshavebeenusedtotreatintestinalailmentsandmildskinconditions.Morerecently,researchhasdemonstratedthatspecificclaysmaypossessantimicrobialproperties.Withthisinmind,wehavefocusedonanewmethodtotreatClostridiumdifficile(C.difficile),theleadingcauseofinfectiousdiarrhoeawithinhospitals,andMethicillin-resistantStaphylococcusaureus(MRSA),thenumberonecauseofhospitalskininfectionsworldwide.Usinggeochemicaltechniques,suchasX-raydiffractionandInductively-coupledplasmamassspectrometry,thephysicochemistryofseventestclayswasdeterminedtoassistinunderstandingtheantimicrobialmechanismoftheclay.Totesttheantimicrobialcapabilityofthetestclays,viabilitycountswereusedwithhydratedclaymineralsandbothantibiotic-susceptibleandantibiotic-resistantpathogenicbacteriatoassessthefeasibilityofusingclaymineralsastherapeuticagents,ie.‘nutraceuticals’.The‘Frenchgreen’clay,composedof91%quartz,demonstratedcompletesterilisationofbothbacteriafollowingovernightincubation;supportingpreviousresearchwithotherpathogenicorganisms.Toestablishtheuseofclaysasgeo-medicaltherapeutics,furtherpharmacotoxicologyusinginvitrohumantissuemodels(i.e.gutandskin)willbeemployedtoelucidatethemechanismsofclaybioreactivity.Thisstudywillhelptofurthertheunderstandingofantimicrobialclays,potentiallyleadingtoalternativetherapiestodecreasethecurrentoverprescriptionofantibioticsandtherisingemergenceofantimicrobialresistance.

ErinMyles1,IsabellaCenteleghe1,LesBaillie2,KellyBéruBé1,TimJones3

1SchoolofBiosciences,CardiffUniversity,Cardiff,UnitedKingdom.2SchoolofPharmacyandPharmaceuticalSciences,CardiffUniversity,Cardiff,UnitedKingdom.3SchoolofEarthandOceanSciences,CardiffUniversity,Cardiff,UnitedKingdom

9

PosterNumber:07AsurveyofantibioticresistanceinentericEscherichiacoliisolatedfromungulatesatazoologicalpark

Abstract

Background:Theantimicrobialresistance(AMR)profileofbacteriaisolatedfromdomesticatedanimalsandfree-rangingwildlifehasbeenstudiedextensively.However,therearefewstudiesdescribingtheAMRprofileofbacteriaisolatedfromcaptivewildanimals.ThisstudyinvestigatedthepresenceofAMRphenotypesincommensalEscherichiacoliandsomeoftheenvironmentalfactorsthatmightaffectAMRdynamicsinthetargetpopulation.

Methods:Inthisstudy,freshlyevacuatedfaecalsampleswerecollectedfrom17speciesofhealthyungulatesatMarwellZooinHampshire,Englandthatyieldedatotalof39commensalE.coliisolates.Antibioticsensitivitywasinvestigatedusingagardiskdiffusionmethods.

Results:Sevenoutof39(18%)E.coliisolateswereresistanttomorethanthreeantibioticclasses,themostcommonpatternofresistancewas:penicillins,tetracyclines,aminoglycosidesandsulphonamides.Noneoftheisolatestestedpositiveforextended-spectrumbeta-lactamase(ESBL)orAmpCactivityusingadiskdiffusionscreeningkit.TheE.coliisolateswerefurtheranalysedusingmulti-locussequencetyping(MLST)whichidentifiedfourpairsofidenticalsequencetype(ST)isolatesand27diversestrains.Reviewofthemedicalrecordsofindividualanimalsshowedprevioususeofpenicillins,sulphonamidesandtetracyclines.Therewasnoapparentspatialclusteringoftheresistanceprofileswithinthezoosuggestingthatresistancegeneswerenotbeingspreadbetweenenclosures.

Conclusion:Studyingtheresistancephenotypesofcommensalbacteriaisausefulindicatorformonitoringtheeffectsofantibioticuseandbiosecuritymeasuresinzoos.

Mary-AnneFrank1,MoritzvanVuuren1,MarkChambers2,RobertoLaRagione2,WillJustice3

1UniversityofPretoria,Pretoria,SouthAfrica.2UniversityofSurrey,Guildford,UnitedKingdom.3MarwellWildlife,Winchester,UnitedKingdom

10

PosterNumber:08ZoonoticantimicrobialresistantCandidaandbacterialspeciesisolatedfromcompanionanimals.

Abstract

Theincreasingappearanceofmicroorganismsshowingresistancetoacollectivearrayofantimicrobialdrugspresentsaseriousthreattopublichealthsafety.Prolongedinfectiousdiseasessuchaspneumoniaandcandidiasisarebecomingmoredifficulttotreat,andoftenfatal,astherapeuticsbecomelesseffectiveagainstdeadlyresistantpathogens.Furthermore,systemicanddermalfungalinfectionshavebecomeincreasinglyprevalentincompanionanimalswheretheyrepresentaroutineproblemforveterinarians.Whereas,fungalinfectionsinhumansarecommonandoftenfatal,byendofthe1990sfungalinfectionshadbecometheseventhleadingcauseofhumanmorbidityresultantfrominfectiousdisease.ThetreatmentandcontrolofinvasivefungalinfectionsinanimalstraditionallyreliedontheuseofamphotericinBwiththeazolesbeingintroducedinthelastdecade.DrugresistanceinfungalspeciessuchasCandidaisanemergingproblemwithresistancetotheseantifungalsacommonoccurrenceininfectedanimals.Indeed,theprognosisforinfectedanimalsremainspoorwithmortalityratesexceeding80%.Studiesconductedaimedtoestablishthetype,frequencyandlevelofresistanceofisolatedspeciesfromprolongedinfectionsincompanionanimals.Samplesofinfection(swabs)wereculturedandanalysedforpathogenicspeciesfollowedbyantimicrobialsusceptibilitytesting.Herewepresentaconcisedetailedsummaryofthetypesofpathogeniczoonoticspeciespresentincompanionanimalsandtheirresistancetocurrenttherapeuticagents.Futureworkaimstodevelopnoveldisinfectantagentsandtherapeuticswhichmaybeusedintheveterinarysectortopreventzoonotictransmission.

Keywords:Resistance,Zoonotic,Candida

ElaineMeade1,MarkAnthonySlattery2,MaryGarvey1,2

1CellularHealthandToxicologyResearchGroup,InstituteofTechnology,Sligo,Sligo,Ireland.2VeterinaryPractice,Manorhamilton,Leitrim,Ireland

11

PosterNumber:09DevelopingphagetherapyforthetreatmentofE.coliinfectionsincompanionanimals

Abstract

Background:Antibioticresistanceisaglobalissuethreateningthefutureofmedicine.Theemergenceofresistanceto"antibioticsoflastresort",suchascolistinviatheMCR-1gene,highlightstheurgentneedforidentificationordevelopmentofalternativeantimicrobialstrategies.Thepossibilityofusingbacteriophagesfortreatingmulti-drugresistantinfectionsinveterinarypracticeneedsexploring.Escherichiacoliisassociatedwithawiderangeofinfectionswithincreasingnumbersofmulti-drugresistantstrainsisolatedinsmallanimalpractice.

Results:TenstrainsofE.coliwereisolatedfromdogsinaveterinaryreferralhospital,allofwhichwereimplicatedinsurgicalwoundinfectionsorUTI’s.E.colistrainswerefoundtobedistributedacross4phylogroups,includinggroupsB2andD,whicharethemajorphylogroupsassociatedwithextra-intestinalinfections.Sevenstrainsweremulti-resistant,showinginvitroresistanceto3ormoreclassesofantibioticsbydiskdiffusiontesting,andall10genomescontainedgenesassociatedwithantibioticresistance,suchaspenicillins,sulphonamides,aminoglycosidesortetracyclines.Caninefaecalsamplesandsoilwereobtainedtodetectandextractlyticphages,usingstandardplaqueassaysagainstacontrolstrainofE.coliandthe10canineisolates.Twolyticphageshavebeenisolatedfromthefirstfaecalsampleandarebeingtestedagainstthepanelofcanineisolates.

Futurework:Thisworkispartofanongoingprojectandisintheearlystagesofobtainingsuitablephagesforfurthertestinganddevelopment.

LucyRhys-Davies,ArnoudvanVliet

UniversityofSurrey,Guildford,UnitedKingdom

12

PosterNumber:10AvianPathogenicandcanine,andhumanuropathogenicEscherichiacoli;Reservoirsofantimicrobialresistance

Abstract

Background:AMRisagrowingpublichealthproblem.Theemergenceofmultidrug-resistanceinextra-intestinalEscherichiacoli(ExPEC)strainsisofparticularconcern.Inhumansanddogs,asubgroupofExPECs(UPECs)areanimportantcauseofurogenitalinfections,whileinpoultrytheAPECsubgroupcancausehighmortality.ThisstudyaimedtocomparetheAPEC,canineandhumanUPECsubgroupsusinggenotypicandphenotypiccharacteristics.

Methods:AMRprofilesof52APEC,98humanand133canineUPECweredeterminedusingdiscdiffusionassays.MICsweredeterminedforcolistin.167strains(55poultry,64canineand48human)wereselectedforgenomesequencingandanalysis.

Results:ArangeofAMRprofileswasobservedoverthe283strainstested.TheprofilesinhumanandcanineUPECweremoresimilartoeachotherthantothatoftheAPEC.ComparativegenomicsbasedoncoregenomeSNPsandinsilicophylogroupPCRshowedthatthemajority(63.4%)ofExPECstrainssequencedwerephylogroupB2.However,phylogroupdistributionpartlyreflectedisolationsource,inthat70.8%humanand84.4%canineUPECbelongedtophylogroupB2with<2%phylogroupCisolates,while32.7%and41.8%ofAPECbelongedtophylogroupsB2,andCrespectively.InterestinglycanineandhumanUPECofphylogroupB2clusteredtogether

Conclusion:TheclusteringofhumanandcanineUPECindicatesapossiblesharedsourceortransmissionrouteforUPEC,whilesourcesofAPECarelikelytobemorediverse.SimilarlyAMRprofilesindicateacloserrelationshipbetweenhumanandcanineUPECthanwithAPEC.

SarahJohns,JonathanBetts,DianeNewell,ArnoudvanVliet,RobertoLaRagione

DepartmentofPathologyandInfectiousDiseases,SchoolofVeterinaryMedicine,UniversityofSurrey,UnitedKingdom

13

PosterNumber:11CompleteUsutuvirusgenomesequencesderiveddirectlyfromclinicalsamplesbyusingamultiplexPCR-basedwholegenomesequencingmethod.

Abstract

Usutuvirus(USUV)isazoonoticmosquito-borneflaviviruswhichemergedinEuropein1996.Sincethen,USUVhasbeenthecausativeagentofepizooticsandsmalleroutbreaksamongwildand/orcaptivebirdsinmanycountries.WehavedevelopedaprotocolcomprisingofanovelmultiplexPCRenrichmentprotocol,optimisedlibrarypreparationmethodsforIllumina,andabioinformaticspipelineforgeneratingconsensusgenomes.ItutilisesmultiplexPCRfortargetedenrichmentofviralgenomesfromsamplescontainingasfewas50genomecopiesperreaction.ThismethodsuccessfullyrecoveredwholegenomicsequencesfromorgansamplesrecoveredfrombirdsthathadnaturallydiedfromUSUVinfectionduringanoutbreakinAustriaandHungarybetween2010and2011,obtainingasequencethatwas99.4-99.7%identicaltopublishedsequencesfromtheoutbreak.Duringvectorcompetencestudies,nativeUKmosquitoes(Culexpipiens)wereinfectedwithanAfricanstrainofUSUV(SAAR-1776)viaabloodmeal.WewereabletoobtainUSUVwholegenomesequencesfromtheinputvirusandmosquitoabdomen.Changesintheconsensuslevelwereidentifiedbetweentheinputvirusinoculumandthevirusretrievedfromthemosquitoabdomen,leadingtoonesynonymousandonenon-synonymousaminoacidchange.ThisnovelmethodprovidesausefultoolforcharacterisingUSUVinfectionsduringoutbreaks,determiningthevirusoriginaswellasthechainsoftransmissionwithinoutbreaks.

MaríadelMarFernándezdeMarco1,LuisM.Hernández-Triana1,KarenLMansfield1,LeighThorne1,SarahLumley1,2,3,DeniseA.Marston1,AnthonyRFooks1,4,NicholasJohnson1,2

1VirologyDepartment,AnimalandPlantHealthAgency,Addlestone,UnitedKingdom.2FacultyofHealthandMedicalScience,UniversityofSurrey,Guilford,UnitedKingdom.3PublicHealthEngland,PortonDown,UnitedKingdom.4DepartmentofClinicalInfection,MicrobiologyandImmunology,InstituteofInfectionandGlobalHealth,UniversityofLiverpool,Liverpool,UnitedKingdom

14

PosterNumber:12Identificationoflineage-specificgeneticmarkersofListeriamonocytogenesbypangenomeanalysis

Abstract

Background:TherapidprogressinDNAsequencingisrevolutionisingmicrobiology,andnowallowsfortheanalysisoflargecollectionsofpathogengenomesequencesforevolutionarypatterns,aswellasthepresenceofmarkersforvirulence,transmissionandantimicrobialresistance.Herewehaveinitiatedananalysisofthepan-,coreandaccessorygenomesof966isolatesofthefoodbornepathogenListeriamonocytogenes.

Objective:ToidentifyL.monocytogenesgenesassociatedwithspecificlineagesandisolationsources.

Results:Atotalof966genomeswerecompared(LineageI(510),LineageII(421),LineagesIII/IV(35)),withtheisolatesrepresentingclinicalisolates(281),foodprocessingsites(341),food(225),animals(82)andenvironment(37).ThepangenomewasidentifiedusingRoary(80%identitywithparalogclustering)andconsistedof2,494coregenesand4,720accessorygenes.GenomesclusteredstronglyontheirrespectiveMLST-clonalcomplexes,withmobilegeneticelements(prophages,plasmids,transposons)representingthemajordifferences.AnalysisoflineageCC6confirmedthereportedassociationbetweendisinfectantresistanceandclinicalisolates,whereasinlineageCC2isolatestwomobileelements,bothencodingLeucine-RichRepeat(LRR)internalin-likeproteinsandregulatoryproteins,werepresentinmostclinicalisolates,whileabsentinfood-processingisolates.

Conclusion:HerewehaveanalysedL.monocytogenesgenomesequencestoidentifygenesassociatedwithclinicalisolatesinlineageCC2,althoughthisassociationwasabsentinlineageCC6,highlightingtheneedtotakegeneticbackgroundintoaccountwhendoingthesecomparativegenomicstudies.TheLRRinternalin-likegenesidentifiedherecouldrepresentcandidatesforfuturevirulencestudies.

ArnoudHMvanVliet1,TomParker2

1UniversityofSurrey,SchoolofVeterinaryMedicine,Guildford,UnitedKingdom.2UniversityofSurrey,Guildford,UnitedKingdom

15

PosterNumber:13TheimpactofhostrestrictionofEscherichiacoliontransmissiondynamicsandspreadofantimicrobialresistance(HECTOR)

Abstract

TheJointProgrammingInitiativeonAntimicrobialResistance(JPIAMR)wasformedin2011by15EuropeanCountrieswiththesupportoftheEuropeanCommission,andnowcomprise26countriesglobally.Itisfunding€65millionofbasicandexploratoryresearchonnewantibiotics,stewardshipofexistingantibiotics,andstudiesandcontrolofthespreadofantibioticresistancebetweenhumans,animals,andtheenvironmentinaOneHealthperspective.

Theprevalenceofantimicrobialresistance(AMR)isincreasingrapidlyworldwide.ThecommensalfloraofhealthyhumansandanimalsisareservoirofAMRencodinggenes,andE.coliinparticularcancarrymultipleresistancefactorsthatareeasilymobilised.HostrestrictionmaybeanimportantdeterminantofthelikelihoodoftransmissionofAMRE.colibetweendifferentreservoirs,suchasbetweenanimalandhumanhosts.

HECTORisamultidisciplinaryprojectthataimstocombinetheresources,infrastructures,andresearchstrengthsofmultiplecountriesinordertoidentifygeneticdeterminantsofE.colihostrestriction,andassesstheassociatedimpactoftheseonAMRtransmissionandprevalence.Multiplemethodsarebeingutilised,includingwholegenomesequencingofalargecollectionofE.coliisolatesfromhumanandanimalsourcesfromdifferentgeographicalareasacrossEuropeandVietnam.Experimentalmodels,suchascontinuousflowchickengutbioreactorsystemsandinvivoanimalmodels,willbeemployedtostudytheroleofhostrestrictiononAMRtransmissionandtheinfluenceofAMRonpathogenfitness.

AmandaSFivian-Hughes1,JennyMRitchie1,HuijunLong1,ArnoudvanVliet1,AniPaloyan2,RebeccaDaines1,JonathanBetts1,SarahJohns1,ConstanceSchultsz3,ChristianMenge4,TorstenSemmler5,MartinBootsma6,HoaNgoThi7,LucasDomínguezRodríguez8,StefanSchwarz9,RobertoLaRagione1

1UniversityofSurrey,Guildford,UnitedKingdom.2SPCArmbiotechnology,Yerevan,Armenia.3AcademicMedicalCentre,Amsterdam,Netherlands.4Friedrich-Loeffler-Institut,Jena,Germany.5RobertKochInstitute,Berlin,Germany.6UniversiteitUtrecht,Utrecht,Netherlands.7UniversityofOxford,Oxford,UnitedKingdom.8VISAVET,Madrid,Spain.9FreieUniversitätBerlin,Berlin,Germany

16

PosterNumber:14IodideCompositionsfortheTreatmentofBovineMastitis

Abstract

Bovinemastitisisacommonandcostlydiseaseaffectingthedairyindustry.Itisaninfectionofthemammaryglandcausedbyaplethoraoforganisms.Duetotheshearnumberofcausativeorganismsandtheirubiquitouspresence,mastitiseradicationisunachievable.Therefore,controlandtreatmentaretheonlyfeasibleoptions.Treatmentreliesheavilyonantibiotics.Economically,mastitiscoststheEU€2billionperannum;butattheindividualfarmlevel,mastitisleadstoreducedmilkquality,reducedmilkproduction,highSCC,andculling.

AtWestwayHealthanovelantimicrobialbasedonperoxidase-catalyzedsystemshasbeendeveloped,leavingnoresiduessoitdoesnotrequireamilkwithdrawalandpermitsadditionofmilktothebulktankduringtreatment.Theantimicrobialtargetsbacterialcellsviathegenerationofthehypoioditeionandexperimentalresultsdemonstrateshowthisionquicklyeradicatesanymicrobespresentbothinvitroandinvivo.InvitroexperimentshavedeterminedtheMICtobecomparabletoantibioticscurrentlyusedtotreatmastitis(6-15mg/ml).Resistancewasnotinducedafterpassagingtestorganismsinsublethalconcentrationsoftheantimicrobial.Inaddition,theantimicrobialdoesnotalterthepHofthemilk.

Thisantimicrobialiscurrentlybeingtestedinvivo,showingthatonceitisadministered,theclinicalsignsofmastitisaresignificantlyreduced,nonegativelong-termeffecthasbeenseen,anditdoesnotnegativelyaffecttheSCC.Overall,theresultsofthisstudyshowexcellentpotentialforanon-antibioticapproachtotreatingbovinemastitis.

ConorLarkin1,CathyAbberton1,MartinGordon1,RuairiFriel1,VincentO’Flaherty1,2

1WestwayHealth,Galway,Ireland.2Microbiology,SchoolofNaturalSciencesandRyanInstitute,NationalUniversityofIrelandGalway,Galway,Ireland

17

PosterNumber:15AntibioticResistancePatternsandGeneInterplayBetweenAnimals,HumansandtheEnvironmentinaHigh-DensityLivestock-HumanPopulationinWesternKenya

Abstract

Antimicrobialresistance(AMR)isaglobalhealthproblem.Antimicrobialsarenolongabletocurebacterialinfectionsowingtothetransferofresistancegenesbetweenbacteria.Thedouble-discdiffusionisthegoldstandardfordeterminingAMRpatterns,however,technologicadvancesinwholegenomesequencing(WGS)hasprovideduswithanothermethodtoinvestigatethediversityofantimicrobialresistancegenes.ThisstudydeterminedresistancepatternsofE.coliisolatedfromhumans,animals,andtheenvironmentfrommultiplestudysitesinWesternKenya.

Faecalsampleswerecollectedfromarepresentativenumberoffarmanimals,farmers,andtheirimmediateenvironments.E.coliwasisolatedandpurifiedfrom680samples.Phenotypicantimicrobialsusceptibilitywasdeterminedinallsamplesbydouble-discdiffusion(EUCAST),followedbyWGS(Illumina,HiSeq)ofasubsetof192E.coliisolates.MLSTwasperformedandresistancegeneswereidentifiedusingResFinder.

HighprevalenceofAMRE.coliwasfoundinfarmer,animal,andenvironmentalsamples;resistancetotetracycline,trimethoprim,andsulfathiazole,wasthemostcommon.WGSanalysisindicatedthatST-196wasthemostcommonST-type.AllST-196E.colihadthesameserotype(O8:H7).Phylogeneticanalysis(usingSNPs)indicatedthattherewasnospecificcross-overoverE.colibetweenhumans,animals,andtheenvironmentwithineachfarm.

Suitablebioinformaticsmethodswhicharerapid,accurateandeasilyinterpretablearestilllacking.TheWGSworkaspartofthisstudywillbecomparedtopreviouslaboratorydatatoaidthevalidationofthismethod.

StevenKemp,NicolaWilliams

UniversityofLiverpool,Neston,UnitedKingdom

18

PosterNumber:16Anthraxandclayminerals:anovelsolution.

Abstract

Bacillusanthracis,thecauseofAnthrax,isendemictolargeswathesofAfricaandSouthAmerica.Itsendosporespersistintheenvironmentandaretheprimarycauseofinfection(toanimalsandhumans).Multiplestateactorsandterrorismgroupshaveutilisedthesporesasbioweapons.Currentmethodsofdecontamination,whilsteffective,causeenvironmentaldecimationandarethemselvesenvironmentallytoxic.Thus,itisdesirabletoconsidernovelmethodsfordecontaminationofaffectedenvironments.

Recently,somebio-reactiveclayshavedemonstratedantimicrobialproperties.GeochemicalinvestigationbyX-raydiffractionandInductively-coupledplasmamassspectroscopyonarangeoftestclaymineralshasbeenundertakentohelpelucidatethemechanismofaction.

AntimicrobialabilitywasassessedusingviabilitycountsofhydratedclaymineralsonbothvegetativecellsandsporesofSternestrainB.anthracis.Oneclay(‘FrenchGreen’)hasshownLog105reductionofviablecoloniesinbothvegetativecellsandspores(0.15g/mland0.05g/ml,respectively)after20-hourincubation.FrenchGreenclayis91%quartzmineralandhasbeenusedasatopicaltreatmentforcutaneousMycobacteriumUlceransinfections.Theseresultssupportpreviousstudiesonotherorganisms.

Furtherin-vitrotoxicologicalstudieswillbeemployedtodeterminetheactivityofantimicrobialclaymineralsonhumantissuemodels;toensurethatdeploymentoftheclayasanenvironmentaldecontaminantwouldnotbehazardoustoinhabitants.

Thisstudywillaidtheunderstandingoftheactionmechanismofantimicrobialclaysandhelpdeterminethepossibilityofclaymineralusageasanenvironmentaldecontaminationsolutionforbacterialcontaminants.

AdamMoore1,ThomasDavies2,LesBaillie3,KellyBéruBé1,TimJones4

1SchoolofBiosciences,CardiffUniversity,Cardiff,UnitedKingdom.2SchoolofBiosciences,CardiffUniversity,UnitedKingdom.3SchoolofPharmacyandPharmaceuticalSciences,CardiffUniversity,Cardiff,UnitedKingdom.4SchoolofEarthandOceanSciences,CardiffUniversity,Cardiff,UnitedKingdom

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