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R. L. Dunkle
RichardDunkle,DeputyAdministrator USDA/APHIS/PPQ
October24,2008
Ralstonia solanacearum r 3 b 2
Credits
Contributors
PhilipBerger,USDAAPHISPPQCenterforPlantHealthScienceand Technology(CPHST),Raleigh,NorthCarolina RussBulluck,USDAAPHISPPQEmergencyandDomesticPrograms, Raleigh,NorthCarolina WilliamCallison(retired),CaliforniaDepartmentofFoodandAgriculture andtheNationalPlantBoard GeneCross,NorthCarolinaDepartmentofAgricultureandConsumer Services JoannCruse,USDAAPHISPPQ,Madison,Wisconsin ReneeDeVries,USDAAPHISPPQCPHST,NationalPlantGermplasm& BiotechnologyLab,Beltsville,Maryland CharlesDivan,USDAAPHISPPQBiologicalandTechnicalServices, Riverdale,Maryland LynnEvansGoldner,USDAAPHISPPQEmergencyandDomestic Programs(EDP),Riverdale,Maryland MikeFirko,APHISPPQPHPPermits,Registrations,Importersand Manuals(PRIM),Riverdale,Maryland LloydGarcia,USDAAPHISPPQEasternRegionalOffice,Raleigh,North Carolina DavidKaplan,USDAAPHISPPQEDP,Riverdale,Maryland LaureneLevy,USDAAPHISPPQCPHST,NationalPlantGermplasm& BiotechnologyLab,Beltsville,Maryland SteveMarquardt,NorthDakotaStateUniversity,Bismarck,NorthDakota WendyNelson,USDAAPHISPPQEnvironmentalServices(ES),Riverdale, Maryland
Ralstonia solanacearum r 3 b 2
Consultants
BillyNewton,USDAAPHISPPQEasternRegionalOffice,Raleigh,North Carolina SusanOToole,USDAAPHISPPQES,Riverdale,Maryland MatthewRoyer,USDAAPHISPPQEDP,Riverdale,Maryland LinSchmale,SeniorDirectorofGovernmentRelations,SocietyofAmerican Florists CliffordSmith,USDAAPHISPPQWesternRegionalOffice,Ft.Collins, Colorado JohnStewart,USDAAPHISPPQEasternRegionalOffice,Raleigh,North Carolina GeraldWheeler,USDAAPHISPPQ,GrandRapids,Michigan JimWriter,USDAAPHISPPQEnvironmentalServices,Riverdale, Maryland RichardZink,USDAAPHISPPQCenterforPlantHealthScienceand Technology,Ft.Collins,Colorado
Consultants
CaitilynAllen,DepartmentsofPlantPathologyandWomensStudies, UniversityofWisconsin,Madison,Wisconsin MargeryDaughtrey,CornellUniversity,PlantPathology,LongIsland HorticultureResearchLaboratory,Riverhead,NewYork JohnElphinstone,DepartmentforEnvironment,FoodandRuralAffairs, CentralScienceLaboratory,York,UnitedKingdom
Cover
GeraniumWiltingsymptomsongeraniumscausedbyRalstonia
solanacearumrace3biovar2;courtesyofWisconsinDepartmentof Agriculture,TradeandConsumerServices
PotatoesSymptomsofbrownrot,Ralstoniasolanacearumrace3biovar2,on
potatotubersandplant;courtesyofCentroInternacionaldelaPapa (InternationalPotatoCenterorCIP),Lima,Peru
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Weed HostBitternightshade,aweedhostofRalstoniasolanacearumrace3
biovar2;courtesyofJoelFloyd,USDAAPHISPPQ
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Ralstonia solanacearum r 3 b 2
Contents
Credits page Ai
Contributorsi Consultantsii Coverii
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Procedure2 GeraniumHosts3 SolanaceousHosts4 GeneralSurvey4 TargetedSurvey4 DelimitingSurvey6 GeraniumHosts6 SolanaceousHosts6 ListofFacilities6 WorkingwithGrowersorFarmManagers7 TracebackAndTraceForwardSurvey7 TracebackSurvey7 AnalyzingInformation7 PositiveDetectioninPotatoes8 TraceForwardSurvey8 WaterSourcesandIrrigationMethods8 InvestigationsatProcessors9 MonitoringSurvey9 GeraniumHosts9 SolanaceousHosts9 VisualInspectionForDetectionSurvey10 GreenhouseandNurseryCrops10 Geraniums10 OtherHostsinGreenhousesorNurseries11 DiscardedPlants11 PotatoCroppingSystems11 PotatoPlants11 PotatoTubers12 TomatoesandOtherSolanaceousCrops13 VisualInspectionForDelimitingSurvey14 GreenhouseandNurseryCrops14 PotatoCroppingSystems14 TuberSampling14 DelimitingSurveysinaGeneralGrowingArea15 SamplingofWaterSources15 TomatoandOtherSolanaceousHosts15 DataCollection15
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DiagnosticScreeningLaboratories6 AlternativeLaboratory6 RequiredPermits6 IntrastateTransfer7 InterstateTransfer8 DiagnosticTestsToDetermineGenusAndSpecies8 ValidatedScreeningTestKits8 ELISATestKits9 WaterTesting9 Culturing9 ConfirmatoryTestingToRaceAndBiovar9 PositiveConfirmations10
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Contents1 Introduction1 InstructionsToOfficers1 QuarantineActionsRequired2 Geraniums2 PotatoCroppingSystems2 EmergencyQuarantineAction2 AccesstoPrivateProperty3 RegulatoryRecords3 OverviewOfRegulatoryProgramAfterAUnitedStatesDetection3 DeterminingOriginofInfections3 Geraniums3 PotatoCroppingSystems4 DistributionofListsofPositiveandSuspectProperties4 Geraniums4 PotatoCroppingSystems5 FurtherInvestigationsAtNurseriesWithSuspectGeraniums6 PlacingHolds7 Nurseries7 PotatoStorageFacilitiesandGrowerFields7 RecordKeeping8 Geraniums8 PotatoCroppingSystems8 PlantstoHold9 SuspectGeraniums9 Variety9 Shipment9 PotentiallyInfectedPlants9 SuspectPotatoCroppingSystems11 GrowerRequirementsUnderQuarantine11 IssuingPPQForm523EmergencyActionNotification11 GeraniumAndPotatoVolunteersAndCullPiles12 HarvestingEquipment,StorageAreas,AndConveyances12 RegulatoryRequirementsOfSeedProducers12 RegulatoryRequirementsForProcessors12 RegulatedArticles13 CultivatedHostPlants13 WeedHostPlants13
Ralstonia solanacearum r 3 b 2
PotsandMedia14 ToolsandEquipment14 RemovingAreasFromQuarantine14 RequirementsForPotatoCroppingSystemsUnderQuarantine15 PropertieswithPositiveTestingorAssociatedFields15 FieldsAdjacenttoPositiveTestingorAssociatedFields,orSharingWater15 StorageFacilities15 VolunteerPotatoesandCullPiles16 HarvestingEquipment,StorageAreas,andConveyances17 RegulatoryRequirementsofSeedProducers17 RegulatoryRequirementsforProcessors17 UseOfChemicals18
Chapter 6 Control
Contents1 Introduction1 LawsPertainingtoPestControl1 ControlRecords2 ControlDecisionsandOversight2 HandlingOfSamples2 PlantDisposalAndDestructionMethods3 BurningorIncineration3 SteamTreatmentUnderPressure3 SoilFumigation3 ApprovedLandfill3 DisinfectionProcedures4 ToolsandEquipment5 ExposedIrrigationSystems5 HoldingPonds5 OutdoorSoilorHoldingAreas5 ApprovedDisinfectants6
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Contents1 PPQForm391SpecimensForDetermination2 AddendumtoPPQForm3914 QuestionnaireForNurseryAndPotatoCroppingSystemOwnersOrManagers5 Location5 Questions5 PPQForm523EmergencyActionNotification7 Instructions8 Procedure8 ReleasingPlants8 DocumentingActionsTaken9 DisposalMemorandum9 PPQForm526PermittoMoveLivePlantPlantsorNoxiousWeeds10 11
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Contents1 Introduction1 StatusofCommercialDiagnosticKits1 Nonplate,Rapid,ImmunoassayDetection2 AdgenSpotCheckLFTM2 AgdiaImmunoStrip2 CSLPocketDiagnosticTM3 ImmunoplateAssays(ELISA)3 AgdiaPathoScreenELISA3 ComparisonOfNonplateImmunoassayAndPlateImmunoassay4 ValidationOfCentralScienceLaboratoryTaqmanAssay5 ValidationOfSampleAndDnaPreparationMethods5 PreparationOfMediaForIsolation6 SelectiveMediumforRalstoniasolanacearum6 SemiselectiveMediumforRalstoniasolanacearum6 IsolatingBacteriaFromTissueSamples8 MethodA8 MethodB8 MethodC8
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Appendix E page E-1 PelargoniumintheNurseryTrade page E1 Appendix F page F-1 ComparisonofSymptoms page F1 Appendix G page G-1 Disinfectants page G1
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Chapter 1
Introduction
Contents
Purpose page 11 Status page 12 DocumentComprehensiveness page 12 CommercialSuppliersOrProducts page 12 PreventionOfInfection page 12 ProgramSafety page 13 SupportForProgramDecisionmaking page 13
Purpose
UseNewPestResponseGuidelines:Ralstoniasolanacearumrace3biovar2asa guidewhendesigningaprogramtodetect,diagnose,contain,control,or eradicatethispathogen.Personnelwithinstatedepartmentsofagriculture andothersconcernedwithdevelopinglocalsurveyorcontrolprograms shouldfindtheGuidelinesuseful. UnitedStatesDepartmentofAgricultureAnimalandPlantHealthInspection ServicePlantProtectionandQuarantine(USDAAPHISPPQ)developedthe Guidelinesthroughdiscussion,consultation,oragreementwithstaffat USDAAgriculturalResearchService(ARS),universities,industries,andstate departmentsofagriculture. TheGuidelineswillbeupdatedasnewinformationbecomesavailable.Specific emergencyprogramsshouldbebasedoninformationavailableatthetimeof theincident.
Recent Updates
Significantchangestothisdocumentasaresultofnewinformationwillbe providedinthissection.Updatestothecurrentversionincludethefollowing: Thediagnosticlaboratorywithproperauthorizationstodofinal confirmatorytestingforRalstoniasolanacearumrace3biovar2isthe USDAAPHISPPQMolecularDiagnosticsLaboratory(MDL)in Beltsville,MD.MDLhasthenecessarypermitsandregistrationsto possessthisselectagent.SeeIdentificationonpage 41. GeographicDistributiononpage 27wasupdated.
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May 12, 2008 Emergency and Domestic Programs Introduction.fm Ralstonia solanacearum r 3 b 2 1-1
Chapter 1 Status
Status
Ralstoniasolanacearumisaplantpathogenicbacteriumthatcauseswilt diseases.StrainsofRalstoniasolanacearumcauseeconomicdamagetodiverse agriculturalcropsandarewidespread(Hayward,1991;Sequeira,1994). Ralstoniasolanacearumrace3biovar2isofparticularconcernintheUnited Statesisbecauseofitsaffectoneggplant,geraniums,potatoes,andtomatoes. Whilerace1isendemictothesoutheasternUnitedStateswhereitcanaffect tomatocrops,Ralstoniasolanacearumrace3biovar2isnotknowntooccurin theUnitedStatesandisconsideredofquarantineimportance.
Document Comprehensiveness
Thisdocumentisnotintendedtobecompleteandexhaustive,butprovidesa foundationbasedupontheliteratureavailabletoassistfurtherwork.Some keyarticleswerenotavailableatthetimeofwriting,andnotallspecialists andmembersoftheresearchcommunitywereconsultedfortheiradvice.
Prevention Of Infection
Tominimizetheeffectsofthisdiseaseandcontrolactionsthatmaybe necessary,regulatoryofficersandgrowersshoulddevelopandimplement effectivesanitationprocedurestoensurethatthepathogendoesnotspread withintheirgreenhouseornurseryfacilities,associatedsupportbuildings, equipmentorvehicles.
Regulatory Officers
Federalandstateregulatoryofficersmustconductinspectionsandapply controlmeasurestoensurethatthediseaseorpathogendoesnotspread withinorbetweengreenhousesornurseries,associatedsupportbuildings, equipment,vehicles,orfields,anddoesnotescapeintootherproduction systems.
Important Since inspectors could inadvertently spread Ralstonia solanacearum race 3 biovar 2 or other pathogens during the inspection process, federal and state regulatory officers conducting inspectionsbefore entering and upon leaving each greenhouse and nursery locationmust follow sanitation guidelines to prevent spreading contaminated plant material, soil, or water to other facilities. See Precautions on page 3-1 for more information on this topic.
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Growers
Geraniumgrowerscanminimizetheimpactofthedisease,includingthe regulatoryimpact,byfollowingthesepractices: Submitfortestingplantsthatdisplaywiltsymptomsanddonotrecover afterwatering. Avoidsubirrigationandsharedwateringsystems. Usestrictsanitarypracticeswhenhandlingorpropagatingplants. Donotplaceplantsbeneaththedriplineofhanginggeraniums. Maintainvarietiesofgeraniumsfromforeignsourcesseparate. Disinfectwatersystemsandregularlytreatirrigationwaterfromany source. Keepgreenhouses,areasaroundgreenhouses,andirrigationwater holdingoroverflowpondsfreeofweeds. Domesticgrowerscanimplementthepracticestominimizecontaminationof rootedcuttingsforsaleintheUnitedStates. GrowerscanconsultthedocumentMinimumSanitationProtocolsfor OffshoreGeraniumCuttingProduction,developedbyUSDAAnimaland PlantHealthInspectionService(APHIS)andusedbyoffshoregeranium suppliers,formoreinformation.
Program Safety
Safetyofthepublic,andprogrampersonnel,isapriorityconsiderationin preprogramplanningandtraining,andthroughoutprogramoperations. Safetyofficersandsupervisorsmustenforceonthejobsafetyprocedures.
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Chapter 2
Pest Information
Contents
Introduction page 21 Classification page 21 Importance page 22 History page 23 EconomicImpact page 26 Hosts page 26 GeographicDistribution page 27
Introduction
UseChapter2PestInformationtolearnmoreabouttheclassification, importance,history,hostrange,anddistribution,ofRalstoniasolanacearum race3biovar2.
Classification
UseTable 21asanaidtotheclassificationofRalstoniasolanacearumrace3 biovar2.
TABLE 2-1 Classification of Ralstonia solanacearum race 3 biovar 2 (continued)
Domain Phylum Class Order Family Strain Synonyms Approved common names (APS, 2003) Bacteria Proteobacteria Betaproteobacteria Burkholderiales Ralstoniaceae Ralstonia solanacearum (Smith 1896) Yabuuchi, et al. 1995 [race 3, biovar 2] Bacillus solanacearum (Smith 1896), Pseudomonas solanacearum (Smith) Smith 1914, Burkholderia solanacearum (Yabuuchi, et al. 1992) bacterial wilt (pepper, potato, tomato), southern wilt (geranium), brown rot (potato)
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Chapter 2 Importance
Importance
Ralstoniasolanacearumisaplantpathogenicbacteriumthatcauseswilt diseases.StrainsofR.solanacearumcauseeconomicdamagetodiverse agriculturalcropsandarewidespread(Hayward,1991;Sequeira,1994) (Table 22).OfparticularconcernintheUnitedStatesisR.solanacearumrace3 biovar2becauseofitsaffectoneggplant,geraniums,potatoes,andtomatoes. Whilerace1isendemictothesoutheasternUnitedStateswhereitcanaffect tomatocrops,R.solanacearumrace3biovar2isnotknowntooccurinthe UnitedStatesandisconsideredofquarantineimportance.
TABLE 2-2 Host Range and Distribution of Strains of Ralstonia solanacearum [Adapted from Daughtrey (2003) and Denny and Hayward (2001)]
Race 1 2 3 4 5 Biovar 1, 3 or 4 1 2 3 or 4 5 Hosts Wide Musa spp. Potato, other Solanaceae, geranium, others Ginger Mulberry Geographic Distribution Asia, Australia, Americas Caribbean Islands, Brazil, Philippines Worldwide except United States and Canada Asia China
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Chapter 2 History
Thepathogenisnotreadilyspreadbetweenplantsbythefollowingmeans: Splashingofwater Leaftoleafcontact Aerialtransfer Infectionoccurstypicallythroughtherootsandwoundinginrootareas,a normalphysiologicalprocessasrootletsgrow.Bacteriaarenormally concentratedinthelowerstemportionsoftheplant.Spreadcanbecontrolled ingreenhousesbythestrictapplicationofsanitationpractices. Thespeciesisdifferentiatedintofiveracesaccordingtohostrange (Buddenhagen,Sequeira,andKellman,1962),andintobiovarclassifications accordingtophysiologicaltests(Hayward,1964;Hawyard,1991).
Race 1
StrainsofRalstoniasolanacearumrace1biovars1and3havebeenfoundto occurongreenhouseornamentalsinthenorthernhemisphere(CABI,2003). Race1alsohasawiderhostrangeandisendemictothesoutheasternUnited States.
Race 3
Ralstoniasolanacearumrace3isfoundworldwideexceptintheUnitedStates andCanada.HostsofR.solanacearumrace3biovar2areusuallyrestrictedto cultivatedsolanaceousspeciessuchaspotatoandtomato.Race3hasbeen reportedoccasionallyoneggplant(Solanummelongena),pepper(Capsicum annum)andsolanaceousweeds(Martin&French,1995).Someinfectedweed hostsremainsymptomless.Theseweedhostsmayenablerace3biovar2to surviveinalatentformwithinthehostorintheirrootareasinthesoil(Janse, etal.2003).
History
Geranium Hosts
IntheUnitedStates,a1979studyofhosttestswithastrainofRalstonia solanacearumisolatedfromPelargoniumxhortorum(=P.zonalehybridsor geranium)indicatedthepresenceofrace3becausetheisolatewasnot pathogenicontobacco(Strider,etal.1981).However,somereportsoffindings ofR.solanacearumrace3biovar2onPelargoniumspp.fromWesternAustralia andTanganyikaarequestionable(Pittman,1933;Wallace,1934). In1995,KimandOlson(2003)receivedastrainofRalstoniasolanacearumrace3 biovar2fromaConnecticutsource.Thestrainwasisolatedfromageranium plantfromGuatemala.Theyconfirmedthepresenceofasimilarstrainon wiltedgeraniumplantsfromgreenhousesinPennsylvaniain1999and2000 andfromDelawarein1999;theplantswereimportedfromGuatemala,too.
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Chapter 2 History
In1999,Ralstoniasolanacearumrace3biovar2wasreportedinNewJersey, NewYork,Ohio,Pennsylvania,SouthDakotaandWisconsinoncommercially growngeranium(Pelargoniumzonale)importedfromGuatemala(Hudelson,et al.1999and2002;Nameth,1999;SPRO,2002).Thesefindingsresultedina reviewofforeignsiteproductionfacilitypracticesthatexportgeranium cuttingstotheUnitedStates. Alsoin1999,thebacteriumwasdetectedintheUnitedKingdomonimported geraniumcuttingsproducedinKenyafortheEuropeanmarket(Janse,etal. 2003).DuringSeptemberDecember2000,symptomsofbacterialwiltwere observedinseveralgeraniumnurseriesinBelgiumandGermany(Janse,etal. 2003). InFebruary2003,geraniumplantsimportedfromKenyatotheUnitedStates wereimplicatedasthesourceofRalstoniasolanacearumrace3biovar2that resultedindetectionsin127individualnurseriesin27states.Importsfrom Kenyawerehaltedandphytosanitaryrequirementsimplementedforall geraniumimportsfromcountriesthathaveR.solanacearumrace3biovar2.In early2004,anotherdetectionwasmadeintheUnitedStates,thistimetraced backedtoproductiongreenhousesinGuatemala.Thisprogram,again conductedbyfederalandstateregulatoryofficials,resultedincontrolactions in453nurseriesin41states.Bothactionsresultedinthedestructionof approximatelytwomilliongeraniumplantseach. In2003and2004,detectionsintheUnitedStateshavebeenlimitedtowithin greenhouses.Theinfectionswerecontainedandplantsweredestroyed.The diseasewaseradicatedthroughindustrypracticesorcontrolprograms.
Current Status
Solanaceous Hosts
In1880,astrainofbacterianowrecognizedasRalstoniasolanacearumrace 1infectedtobaccoinGranvilleCounty,NorthCarolina,causingsymptoms ofbacterialwilt1.Thediseasebecamesoseverethatgrowersexperienced lossesof25100%,andtheeconomiesofentirecommunitiesweredestroyed (MeltonandShew,2000).Atthattime,thediseasewasnamedGranvillewilt forthecountyinwhichitwasfirstobserved.Aroundthesametime,an unidentifieddiseasebegantoaffectsouthernpotatogrowersandtomato growersinMississippi(Kelman,1953).
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Chapter 2 History
Whenscientistsfirstrecognizedbacterialwiltsofsolanaceouscrops,they madenoattempttodifferentiatebetweenracesorstrains.Thehostdifferential testtoseparateRalstoniasolanacearumraces1,2,and3,wasdevelopedinthe 1960s;limitedinformationregardingracedesignationwasavailablebefore thattime. Sincethe1950s,scientistshaverecognizedthatinfectionsofRalstonia solanacearumraces1and3havehinderedmanysolanaceouscropsinboth tropicalandtemperateregions.Race1hascausedthegreatesteconomic damagetopotato,tobacco,andtomatocropsinthesoutheasternUnited States.InIndonesia,Brazil,Colombia,andSouthAfrica,races1and3caused thegreatesteconomicdamage(CABI,2006).InthePhilippinesduringthe period19661968,races1and3causedlossesof15%,10%,and25%,in tomato,pepper,andtobaccocrops,respectively(Zehr,1969). InIndia,infectionoftomatoeswithRalstoniasolanacearumraces1and3can resultinthecompletelossofacrop.InIsrael,Volcani&Palti(1960)reported thatpotatolossesweregreaterinthespringcropthantheautumncropdueto hightemperaturesduringtuberformationandmaturity.Extensivelossesof potatowerereportedinGreeceduringtheperiod19511953(Zachos,1957).
Current Status
Today,Ralstoniasolanacearumrace1isendemicinsomeareasofthesouthern UnitedStates,wheremanagementofthediseasehasreducedlossesto12% ofproductionor$1015millionannuallyintobacco,withsimilarlossesin tomatoandpepper. Ralstoniasolanacearum,thecausalagentofpotatobrownrot,hasbeen describedasoneoftheworldsmostdestructiveplantbacterium.Ithas affectedatleast3.7millionacresofpotatoesinmorethan80countries,with estimatedannuallossesofmorethan$950million(DEFRA,2003a).Losses duespecificallytoR.solanacearumrace3biovar2cannotbequantifiedsince reliabledataareunavailable.Nevertheless,intheearly1990sthispathogen causedseriouslossesintheEuropeanpotatoindustry. Duetorecentoutbreaks,insuranceclaimsintheNetherlandsin1998and 1999asaresultofpotatobrownrotcausedbyRalstoniasolanacearumrace3 biovar2wereestimatedatU.S.$2.7million(Meiners,2000).Inaddition,the U.S.floricultureindustryincurredsubstantiallosseswhenthebacteriumwas recentlydetectedandconfirmedingeraniumcuttingsimportedfromKenya andGuatemala.Foralimitednumberofnurseries,thecostofdisposaland replacementofrootedgeraniumplugsduetorace3biovar2wasestimatedto bearound$221,632,withtotallossesfor70nurseriesof$1.4million(L.Garret, USDAAPHISCPHSTeconomist,personalcommunication).However,the totalnumberofestablishmentsthatsufferedlossesduetoR.solanacearumrace 3biovar2isunknownatthistime,thusaccuratelossesduetothisbacterium cannotbecalculated.
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Economic Impact
Geranium Hosts
LikeXanthomonaspelargonii,infectionbyRalstoniasolanacearumrace3biovar2 cancausemortalityingeraniums.However,growerscanmanageRalstonia solanacearumbycullingandfollowingpropergreenhousesanitationpractices. TheeconomicimpactofRalstoniasolanacearumrace3biovar2ongrowers, importers,anddistributorsintheUnitedStatesisduetotheregulatory actionsnecessarytoeradicatethediseaseoncedetected.
Solanaceous Hosts
Ralstoniasolanacearumrace3biovar2isaseriouspathogenthatcausesbrown rotorbacterialwiltofpotato(Hayward,1991;Janse,1996).Bacterialwiltof potatohasbeenestimatedtoaffect3.75millionacresin80countries.Annual globaldamageestimatesexceed$950million(DEFRA,2003a).Itisadaptedto coolertemperaturesandwouldbeparticularlydamagingtopotato productionregionsoftheUnitedStates. Whilerace1causeslossestotomatocropsinFlorida(Momol,etal.2003),the affectofrace3ontomatoesandothersolanaceouscropsintheUnitedStatesis unknown.Ralstoniasolanacearumrace3biovar2causesmortalityingeranium plants,butXanthomonaspelargoniiisagreaterconcernofgrowers.However, theregulatoryaffectcanbemoresignificantforgrowerswiththediseasein theirfacility.
Hosts
Excludinggeranium(Pelargoniumspp.),mosthostsofRalstoniasolanacearum race3biovar2areinthefamilySolanaceae.Themajorcrophostplantspecies includepotato,tomato,peppersandeggplant.Othercrophostshavebeen implicatedbylaboratorytesting.WeedhostsincludemanySolanumspp.See Janse,etal.(2003)foracompletereviewofhosts.SeePelargoniuminthe NurseryTradeonpage E1formoreinformationonnamingofgeraniums.
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Geographic Distribution
Geographicdistributionrecordslistedbelowareofconfirmedrace3orof presumedrace3basedonpathogenicitytopotatoandtomato(CABI,2003). North AmericaMexico(CABI,2003). EPPO RegionBelgium,Estonia,France,Germany,Hungary,Ireland, Netherlands,Latvia,Lebanon,Lithuania,Moldova,RussianFederation, Slovakia,Slovenia,UnitedKingdom,FormerYugoslavia,FormerUSSR (CABI,2003). AsiaBangladesh,China(Fujian,GuangdongGuangxi,Hebei,Jiangsu, Taiwan,Zhejiang),India(HimachalPradesh,MadhyaPradesh,Maharashtra, Manipur,Meghalaya,TamilNadu,Tripura,UttarPradesh,WestBengal), Indonesia(Java),Iran,Japan(Kyushu),Nepal,Pakistan,Philippines,SriLanka (CABI,2003),RepublicofKorea(NIAST,2002),Malaysia(Possier,etal.,1999), Taiwan,Thailand(HoritaandTsuchiya,2001). AfricaBurundi,Egypt,Ethiopia,Kenya,Libya,Nigeria(Cook,etal.1994), Runion,Rwanda,SouthAfrica,Uganda(Hayward,1964;Tusiime,etal.1998), Zaire,Zambia,Zimbabwe(CABI,2003). South AmericaArgentina,Bolivia,Brazil(Goias,Parana,Pernambuco,Rio GrandedoSul,SantaCatarina,SaoPaulo),Chile,Colombia,Peru,Uruguay, Venezuela(CABI,2003). Central America and CaribbeanCuba(French,1998),CostaRica, DominicanRepublic,ElSalvador(French,1998),Guadeloupe,Guatemala, Haiti,Honduras,Nicaragua,Panama,WestIndies(CABI,2003). OceaniaAustralia(NewSouthWales,SouthAustralia,Victoria)New Caledonia,PapuaNewGuinea(CABI,2003).
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Chapter 3
Survey Procedures
Contents
Introduction page 31 Precautions page 31 DetectionSurvey page 32 DataCollection page 315 DelimitingSurvey page 36 TracebackAndTraceForwardSurvey page 37 MonitoringSurvey page 39 VisualInspectionForDetectionSurvey page 310 VisualInspectionForDelimitingSurvey page 314
Introduction
UseChapter3SurveyProceduresasaguidetoconductingasurveyforRalstonia solanacearumrace3biovaringeraniums,potatoes,andothersolanaceous hosts. ConductadetectionsurveytodetermineifRalstoniasolanacearumrace3biovar ispresentorabsentinadefinedgreenhouse,nursery,orfieldarea.Aftera newdetectionintheUnitedStates,orwhenadetectioninanewareais confirmed,conductadelimitingsurveytodefinethegeographiclocationof thedisease.Conductamonitoringsurveyifyouhaveappliedacontrol procedureandneedtomeasureitseffectiveness.
Precautions
Whenvisitingsitestoconductsurveysorcollectsamples,regulatoryofficials musttakestrictmeasurestopreventcontaminationbyplantpathogens betweengreenhouses,nurseries,andpotatofacilitiesduringinspections. Acceptablemethodsofprotectionincludetheuseofaneffectiveantimicrobial soap,lotion,ordisinfectantusedaccordingtothelabelinstructions,or disposableglovesandbootieschangedbetweenvisits.SeeDisinfection Proceduresonpage 64formoreinformationondisinfectionoftools, irrigationsystems,ponds,andsoil.
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Detection Survey
Thepurposeofadetectionsurveyistodetermineifapestexistsinanarea. Thiscanbeextremelybroadinscopeaswhenassessingthepresenceofthe diseaseinawideareaoritcanberestrictedtodiscoveringifaspecificpestis presentinacertainarea. Basedstrictlyonanegativeresultinadetectionsurvey,itisnotvalidtoclaim thatapestdoesnotexistinanareaifresultsarenegative.However,negative resultsarevaluableforprovidingcluesastomodeofdispersal,temporal occurrence,orindustrypractices,particularlywhenconsideredwithresults fromsimilarareasorproximities.
Important Detection surveys for geraniums infected by Ralstonia solanacearum race 3 biovar or other cultivated hosts in nurseries should be conducted by state inspectors in conjunction with federal PPQ inspectors. PPQ inspectors may inspect nurseries without a state inspector if they have permission of the nursery owner or manager and have advised the state of their visit.
Procedure
FollowthisprocedurewhenconductingadetectionsurveyforRalstonia solanacearumrace3biovar2: 1. Usevisualinspectiontoexaminecultivatedhostplants(geraniums, potatoes,andothersolanaceoushosts)andsolanaceousweedsfor wiltingsymptoms.Besuretoincludeplantsinthefieldthatwere startedingreenhousesornurseriesinassociationwithimported geraniums.SeeVisualInspectionForDetectionSurveyonpage 310 formoreinformationoninspectionprocedures.
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If: Samples test positive for the species Ralstonia solanacearum by ELISA or culture
And: Their origin is in the southern areas of the United States where race 1 occurs1
Then: Conduct monitoring and further tests at a qualified, authorized diagnostic laboratory to determine race and biovar. See Monitoring Survey on page 3-9 and Identification on page 4-1 for more information.
Samples test negative for the species Ralstonia solanacearum by ELISA or culture
1
While race 1 of Ralstonia solanacearum occurs in the southeastern United States on many of the same hosts as race 3, it is indistinguishable from race 3 without further diagnostic testing.
Geranium Hosts
Usevisualinspectiontoconductadetectionsurveyingreenhouseand nurseryfacilities.SeeVisualInspectionForDetectionSurveyonpage 310 formoreinformationonthistopic. Randomsamplingofgeranium(andpotato)plantslackingsymptomsisnot recommendedatthistimebecauseofalackofsamplingmethodologyand processingcapability.Duetolatency,infectedplantsmaynotshow symptoms.Thereiscurrentlynoreliabletestforasymptomaticplants, thereforesymptomologyisthemostreliableindicatorofpossibleinfection.1
May 12, 2008 Emergency and Domestic Programs Ralstonia solanacearum r 3 b 2 3-3
Solanaceous Hosts
Usevisualinspectiontoconductadetectionsurveyinsolanaceoushosts. DetectionsurveysforRalstoniasolanacearumrace3biovar2inpotatogrowing areascanbegeneral,ortargetedtoareasconsideredtobeatgreaterrisk.See VisualInspectionForDetectionSurveyonpage 310formoreinformation.
General Survey
Targeted Survey
visualobservationsofwiltintomato,pepper,andeggplantcropscanbeused indetectionsurveys.Conductsurveysoftomatoandothersolanaceoushosts if: Hostsweregrowninthesameproductiongreenhousesasimported geraniums,sincetheymayrepresentahighriskpathwayfor introducinginfectionintoafieldorhomegarden. Fieldsarelocateddownstreamornearnurseryfacilitiesthathad importedgeraniumswhichtestedpositiveforRalstoniasolanacearum race3biovar2,orhadvarietiesofsuspectgeraniumsthatwere associatedwithimportedgeraniumsthattestedpositiveforthe pathogen.
Tomato TransplantsTomato,eggplant,andpepperplantsfor
commercialproductionareoftengrownastransplantsinonearea andshippedtoanotherareaforfieldplanting.Theseavenuesof
If available, testing of asymptomatic potato tubers might prove to be more reliable. Latent infections may be present for several years before field symptoms become visible. In the absence of symptoms, if a positive detection occurs in potato seed certification screening, normal traceback or trace forward investigations will indicate potential high-risk sites to survey and test. A potato sampling scheme over a large area may be the most efficacious method of sampling.
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biovar2tospreadinpotatogrowingareasisthroughinfectedtubersusedfor seed.Seedtubersareregularlysampledandscreenedasapartofpotatoseed certificationprograms.Theseprograms,conductedatvariousinstitutions undertheauspicesofstateuniversityorstatedepartmentsofagriculture, assurethatcommercialpotatoseedtubersareinspectedforavarietyofpotato diseasesbeforereceivingcertification. Seedpotatoinspectorsregularlylookforbacterialringrot(Clavibacter michiganensesubsp.sepedonicus).LikeRalstoniasolanacearumrace3biovar2, diagnosticlaboratorytestsareanintegralpartofconfirmingthepresenceof bacterialringrotinsymptomatictubers.Ifdiagnostictestingisperformedto confirmthepresenceorabsenceofbacterialringrot,testsshouldalsobe conductedtoruleoutthepossibilitythatR.solanacearumrace3biovar2isthe causalagent.
Solanaceous Weed HostsRalstoniasolanacearumrace3biovar2cansurvive
inwaterwaysandonweedhosts,includingblacknightshade(Solanumnigrum L.)andbitternightshade(S.dulcamaraL.)(Figure 31).Forexample,bitter nightshadegrowinginriverwaterinEuropeactedasasymptomless, secondaryhostinwhichthepathogenoverwinteredandmultipliedtofurther contaminateirrigationwatersupplies(Janse,1996).Inconjunctionwith detectionandmonitoringsurveys,collectpotentialweedhostsandsendto theappropriatediagnosticlaboratory,aswithcultivatedhostsofR. solanacearumrace3biovar2.Inparticular,weedhostsshouldbecollected aroundholdingpondsandrunoffareasinornearfieldswithhoststhathave testedpositive.
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FIGURE 3-1 Bitter Nightshade, a Weed Host of Ralstonia solanacearum race 3 biovar 2 [Image courtesy of Joel Floyd, USDAAPHISPPQ]
Delimiting Survey
AfteranewdetectionintheUnitedStates,orwhenadetectioninanewareais confirmed,conductadelimitingsurveytodeterminetheextentofthe infestationorsuspectareasinwhichtoconductfurtherinvestigations.
Geranium Hosts
Commercialproductionofgeraniumsislimitedtogreenhouses,nurseries, andretailgardencenters.Lookforplantsconnectedtocirculatingirrigation systems,waterrunoff,andcollectionponds,todetermineiftheinfectionhas spreadtonewareasonoroffthegrowerspremises. Tracebackandtraceforwardinvestigationswillhelpdeterminenewfacilities needinginspection.Developlistsofpotentiallyinfectedfacilitieswithinand outsideastate.Theselistsarenottobesharedoutsideregulatorycooperators. Iftraceforwardstoresidencesfrompositivenurseriescanbeidentified,then thesegeraniumsshouldalsobeinspectedwhenpossible.
Solanaceous Hosts
List of Facilities
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1. Requestthatthegrowerormanagerproduceanydocumentation includinginvoices,shippinglistsassociatedwiththepositivetested suspecttubersonthelistoffacilities. 2. Makecopiesoftherecords. 3. Locateanysuspectmaterialstillonthepremises. 4. Takephotographsoftheareawheresuspectplantmaterialislocated. Includephotographsdepictingconditions,includingtheproximityof neighboringplants,thatmayindicatebroadercontamination. 5. Makearecordofthenumberofplants,type,andlocationincludingan accountingofsuspectplantsfromthatshipmentnotonthepremises. 6. Inspecttheareaforthepresenceofcullpilesanddetermineifany harvesting/processingequipmenthasbeensharedwithotherfarms.
Traceback Survey
Analyzing Information
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Investigations at Processors
taketuberandwatersamplesatfacilitiesthathandledsuspectandpotentially infestedtubers.
Processing EnvironmentDeterminetheprocessingmethods,temperatures
Monitoring Survey
Conductamonitoringsurveyifyouhaveappliedacontrolprocedureand needtomeasureitseffectiveness.SeeControlonpage 61formore informationoncontroloptions.
Geranium Hosts
Onceanurseryhasdestroyedallsuspectgeraniumsanddisinfectedgrowing areas,followupmonitoringisnecessary.Besuretofollowsanitation proceduresduringnurseryvisitsinamonitoringsurvey. Visualinspection,duringthesameorsubsequentgrowingseason,is appropriateinordertoexaminehostsforsymptoms.Ifsamplingprotocolsare available,samplingofwaterfromirrigationsystemsorpondsmayalsobe includedwithadditionalinspectionsofhostcropareasinrunoffareas adjacenttopositivetestingnurseries.SeeVisualInspectionForDetection Surveyonpage 310andVisualInspectionForDelimitingSurveyonpage 314formoreinformationoninspection.
Solanaceous Hosts
OnceRalstoniasolanacearumrace3biovar2hasbeenconfirmedfroma particularfield,andinfectedandpotentiallyinfectedplantshavebeen destroyed,followupmonitoringwillbenecessary.Usethefollowingtools: Visualinspectioninthefield
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Discarded Plants
Conductvisualsurveysofpotatofieldsduringperiodswhentemperatures arehighandsymptomsofbacterialwiltwillbemostobviousifpresent.
EarlySymptomsofbacterialwiltinmaturepotatoplantsthatresultfrom
FIGURE 3-2 Left: Wilted Potato Plant with Symptoms of Brown Rot, Infected by Ralstonia solanacearum race 3 biovar 2. Right: Healthy Potato Plant [Image courtesy of Centro Internacional de la Papa, Lima, Peru] LateMoreadvancedinfectioncanresultinthefollowingsymptoms:
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sepedonicus,occursinpotatogrowingareasoftheUnitedStatesandisanother bacterialdiseaseofpotatothatcauseswiltsymptoms. Thesymptomsofbacterialringrotdifferfrombacterialwiltbyoccurring typicallyontheuppermostleavesorflowerontheplantfirstandcausinga slightrollingatthemarginswithsomelossofcolor.Astheinfection progresses,leaftissuebetweenveinsturnsyellow.Eventually,entireleaves andstemsbecomeyellow,brittle,anddie.Infectioncanbelimitedtooneor twostemsonahill. Whilethesymptomologyofthesetwobacterialdiseasesofpotatomaybe different,thepathogensmustbediagnosedwithlaboratorytesting. Additionally,inareasofthesouthernUnitedStateswhereRalstonia solanacearumrace1exists,asamplingofwiltedplantspositiveforR. solanacearumwillrequirefurthertestingtotheraceandbiovarlevelat qualified,authorizeddiagnosticlaboratories.
Other DiseasesOtherdiseases,aswellasdrought,cancausesymptomsof
Intubersforseedorconsumption,symptomsofRalstoniasolanacearumrace3 biovar2infectionincludethefollowing: Grayishbrowndiscolorationcanbeseenthroughthetuberperiderm. Bacterialoozeaccumulatesaroundthetubereyesoratthestolonend. Soiladherestoooze. Inmoreadvancedlevelsofinfection,squeezingtubermaycausemore bacterialoozetobeexuded(Figure 33). Distinctgrayishbrownvascularringisvisiblewheninfectedtubersare cutincrosssections;canextendintothepithorcortexofthexylem tissue(Figure 33). Applyingpressuretotuberscutinhalfforcesgrayishwhitedropletsof bacterialslimeoozefromthevascularring.
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Infectedtubersleftinthesoilcontinuetodecayandsecondary organismsinvadetobreakthemdown.
FIGURE 3-3 Symptoms of Brown Rot on Potato Tubers Exhibiting External Discoloration and Internal Infection with Ooze Exuding around the Vascular Ring of a Tuber cut in Cross-section [Image courtesy of Centro Internacional de la Papa, Lima, Peru]
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Eachtuberisasamplingunit.Repeatthesamplingforeachfield,warehouse, orstorageunitatagivenlocation.Collect300samplesinordertodetect Ralstoniasolanacearumata1%infestationlevelwith95%confidence.Collect samplesthroughouttheentirebuildingorstoragearea.Iftubersareinbags, collectthe300samplesfromdifferentbags.Disinfectthesamplesthesamples beforelaboratorytesting.SeeIdentificationonpage 41formore informationonlaboratorytesting.SeePotatoStorageFacilitiesandGrower Fieldsonpage 57formoreinformationonpositivetestingseedlotsorfields.
Delimiting Surveys in a General Growing Area
Constructdelimitingsurveysinanareabasedonknownpositivetesting, associatedpositivetesting,orpotentiallyinfestedareasfrominvestigations ofdistributionchannelsandsharedirrigationwater.However,itmaybe necessarytodorandomsamplesinageneralgrowingareatodetectnew infestationsnotdiscoveredthroughinvestigations. Thedelimitingsurveyinageneralgrowingareacanincluderandom samplingofstoredtubersandfieldsthroughoutageographicalarea,with moreintensivesamplingnearknowninfestations.Asthedistanceawayfrom theepicenterofknowninfestationincreases,decreasetherateofrandom sampling.Basedontheepidemiologyandgrowerpractices,anevaluationof riskandresourcesavailablewillhelpdeterminetheextentoftheserandom samplingsurveys.
Data Collection
Surveyorsvisitingsitestoplaceholdsortakesamplesshouldcollectthe followinginformation: Dateofcollectionorobservations Collectorsname Growersfieldorgreenhouseidentificationnumbers
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GPScoordinates Varietyofhostplantgrown Methodsofirrigation Historyoffarmmachineryusage Observationsofwiltandanyotherrelevantinformation Intheabsenceofinspectionofficials,takethefollowingactionsimmediatelyif wiltingsymptomsarenoticed: 1. Markthelocation. 2. Removetheplantsfromthebenchorrack,orflaglocationinfield. 3. NotifyastateorPPQinspector. 4. Placethewholeplantinsidetworesealableplasticbags. 5. Labelthesealedbagswiththefollowinginformation: Date Nameofpersonresponsible Locationofsamplecollection 6. Keepbaggedplantscoolorrefrigerateduntiltheinspectorarrives.Do notfreezethesample.
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Ralstonia solanacearum r 3 b 2
Chapter 4
Identification
Contents
Introduction page 41 Authorities page 41 ProcedureForSamplingAndIdentification page 43 DiagnosticScreeningLaboratories page 46 RequiredPermits page 46 DiagnosticTestsToDetermineGenusAndSpecies page 48 Culturing page 49 ConfirmatoryTestingToRaceAndBiovar page 49 PositiveConfirmations page 410
Introduction
UseChapter4IdentificationasaguidetoidentifyRalstoniasolanacearumrace3 biovar2.Accurateidentificationofthispathogenispivotaltoassessingits potentialrisk,developingasurveystrategy,anddeterminingtheleveland mannerofcontrol.
Authorities
PPQNationalIdentificationServicesMolecularDiagnosticsLaboratory (MDL),Beltsville,Maryland,mustpositivelyidentifybacterialinfectionsto thelevelsofraceandbiovarbeforeinitiationofanyprogramquarantine activities.MDLalsohasthenecessaryregistrationsandcontainment approvalstohandleselectagents.Stateandcooperatinguniversitydiagnostic laboratoriesarelimitedtodeterminationsofbacterialinfectionstothelevelof genusandspeciesRalstoniasolanacearum.SeeDiagnosticLaboratorieson page D1foralistoflaboratoriesandtheiraddresses. PPQpermitandregistrationrequirementsforplantdiseasesandlaboratories fallundertwoauthorities: PlantProtectionAct(7CFRPart330)(SeePlantProtectionActon page 42) AgriculturalBioterrorismProtectionActof2002(7CFRPart331)(See AgriculturalBioterrorismProtectionActonpage 42) Laboratoriesreceivingsuspectinfectedplantmaterialorculturesarerequired tohavePPQpermits.Laboratoriespossessing,using,ortransferringselect agentsarerequiredtoberegistered.However,diagnosticlaboratoriesthat identifyselectagentsareexemptfromthisrequirementaslongasthey
May 12, 2008 Emergency and Domestic Programs Identification.fm Ralstonia solanacearum r 3 b 2 4-1
Chapter 4 Authorities
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Foreachsample,assignandrecordauniqueidentificationnumberinthe followingformat:
Example XX-ABC-0001 where XX is your two-letter state code; ABC is a three letter, state assigned facility code; and 0001 is the sample number for that facility.
Useauniquesamplenumberforeachsample.Keepalogofassignedsample numbers. IfthediagnosticscreeninglaboratoryisapartoftheNationalPlantDiagnostic Network(NPDN),samplesmaybesubmittedusingaNPDNnumberformat. Ifabarcodedsamplenumbersystemisavailable,besuretoincludethe barcodeinthesampleonthePPQForm391.SeeTable D1onpage D10for alistoflaboratoriesintheNPDN. Inspectorsmustprovidethesampledatewithallrelevantcollection informationtotheirStatePlantRegulatoryOfficialorStatePlantHealth Directorassoonaspossible.Thisinformationshouldbecommunicated withinastateandwiththeregionalofficeprogramcontact.Ifasample trackingdatabaseisavailableatthetimeofthedetection,entercollection informationinthesystemassoonaspossible.
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Saturday Delivery
OnlywithspecificpermissionfromAPHISPPQheadquartersshould samplesbesentonFridaysbyFedEx.Althoughitispossibletoarrange SaturdaydeliverybyovernightcarrierstoMDL,oftensamplesarenot deliveredevenwithspecialarrangementwithFedEx.However,MDLwill operateonSaturdayifalargeprogramisbeingconductedrequiring additionalhoursofoperation.IfyouhaveobtainedpermissionfromAPHIS PPQheadquarterstosendsamplesforSaturdaydelivery,pleasesendthe FedExtrackingnumbertoMDLviaemailtoMaryPalmsoshecannotifyher localFedexofficetoauthorizeSaturdaydelivery.
Address Dr. Mary E. Palm USDAAPHISPPQNational Identification Services Molecular Diagnostics Laboratory BARC-East, Bldg. 580 Powder Mill Rd. Beltsville, MD 20705 T: 301-504-7154 or 504-5700 ext. 327 F: 301-504-6124 Email: mary.palm@aphis.usda.gov
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Alternative Laboratory
LaboratorieswithintheNationalPlantDiagnosticNetworkcanscreento genusandspeciesforstatesthatprefernottoorcannotscreentheirown samples.AnysampletestingpositiveforRalstoniasolanacearumwillbe forwardedtoPPQNationalIdentificationServicesMolecularDiagnostics Laboratory(MDL)inBeltsville,Maryland,forconfirmatorytesting.SeePlant DiagnosticNetworksandPrivateLaboratoriesonpage D10toidentifythe regionallaboratoryforyourstate. Statesthatcannotscreentheirownsamples,orhavenotbeenabletomake arrangementswithanotherpermittedlaboratoryoutofstate,canutilize Agdia,Inc.Thisprivatelaboratoryhasthenecessarypermitsand authorizationstoscreentospeciesbutnotraceorbiovar.Samplestesting positiveforRalstoniasolanacearumwillbeforwardedtoMDLforconfirmatory testing.
Address Agdia, Inc. 30380 County Road 6 Elkhart, IN 46514 T: (574) 264-2014, or 1-800-62-AGDIA Web site: http://www.agdia.com
Otherprivatelaboratorieswillbeidentifiedaspermitsareapproved.
Required Permits
Ifastate,university,NationalPlantDiagnosticNetwork(NPDN),orprivate laboratoryperformsaPCRassayanddetectsapresumptivepositiveoutside thenationalsurveyprocessoutlinedhere,theymustabidebythe
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requirementsundertheAgriculturalBioterrorismProtectionActof2002(7CFR 331).Formoreinformation,seeAgriculturalBioterrorismProtectionActon page 42. Diagnosticscreeninglaboratoriesreceivingsamplesaretocommunicatethe dateofreceiptwiththeirStatePlantRegulatoryOfficialand/orStatePlant HealthDirector.Allrelevantsampleinformation,andthediagnostic laboratorysdeterminations,mustbecommunicatedassoonaspossible withinastateandwiththePPQregionalofficeprogramcontact. AnylaboratorythatdiagnosessamplestoRalstoniasolanacearumrace3biovar 2isrequired,undertheAgriculturalBioterrorismProtectionActof2002,to immediatelynotifytheAPHISSelectAgentProgramregardlessofwhethera PlantPestPermitisrequired.
Address APHISSelect Agent Program 4700 River Road, Unit 2 Riverdale, MD 20737 Email: Agricultural.Select.Agent.Program@aphis.usda.gov Telephone: 301-734-5960 Fax: 301-734-3652
Intrastate Transfer
Forinspectorsshippingplantpestsamplematerialtootherlabstotestforthe presenceofRalstoniasolanacearumidentificationtospeciesonly,notbiovar andracenopermitisrequiredforshipmentwithinthestate.Persons shippingplantmaterialtoadiagnosticlaboratoryintheirownstatedonot needtoobtainaPPQpermitundereitherthePlantProtectionAct(Regulations at7CFRPart330)orPPQForm526. WithinstatetransfersofknownRalstoniasolanacearumrace3biovar2are regulatedundertheAgriculturalBioterrorismProtectionActof2002 (Regulationsat7CFRPart331).However,thesubmissionofunknown samplesorthoseonlyidentifiedtoR.solanacearumnottoraceand biovararenotregulatedunderthisAct.
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Interstate Transfer
Stateswithouttheirowndepartmentofagricultureorcooperatinguniversity laboratoryforscreeningtogenusandspeciesmayshipacrossstatelinesto useanotherstateslaboratory. Whenshippingacrossstatelines,theoriginatingstatemustassurethatthe laboratoryinthedestinationstatehasthenecessaryplantpestpermitto receiveinterstatesamples.ApplyforthepermitbycompletingPPQForm523 EmergencyActionNotificationonpage B7. ForthepurposeoftheAgriculturalBioterrorismProtectionActof2002 registrationsarenotnecessary.Aswithintrastatesubmittedsamplesto diagnosticlabs,interstateshipmentofunknownpotentiallyinfectedplantsis notsubjecttoregulationsunderthisAct.
Address
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Chapter 4 Culturing
Address
Ralstonia solanacearum SPOTCHECK LF Adgen, Ltd. Nellies Gate, AYR Scotland, KA6 5AW Web site: http://www.adgen.co.uk T: 44 1292 525275 F: 44 1292 5255477
Water Testing
MethodsoftestingwaterforthepresenceofRalstoniasolanacearumrace3 biovar2havebeendevelopedbyNPGBL,Beltsville,Maryland,butprotocols haveyettobeincludedintheGuidelines.
Culturing
Diagnosticlaboratoriescanforwardculturesafterscreeningtogenusand specieshasbeenperformed.SeeDiagnosticValidationsandIsolation Methodsonpage C1forrecommendedmethodsofculturing.Whensending culturesforraceandbiovardetermination,thesendermustassurethat packagingissecureaswithplantsamples.
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Address
Dr. Mary E. Palm USDAAPHISPPQNational Identification Services Molecular Diagnostics Laboratory BARC-East, Bldg. 580 Powder Mill Rd. Beltsville, MD 20705 T: 301-504-7154 or 504-5700 ext. 327 F: 301-504-6124 Email: mary.palm@aphis.usda.gov
Positive Confirmations
IncompliancewiththeAgriculturalBioterrorismProtectionActof2002(7CFR Part331)ifadiagnosticlaboratoryheldbackpartofascreenedsampleor cultureforvoucherpurposesandthesampleforwardedtothePPQNational IdentificationServicesMolecularDiagnosticsLaboratory(MDL),Beltsville, Maryland,camebackaspositiveforRalstoniasolanacearumrace3biovar2,the diagnosticlaboratoryisrequiredtonotifyPPQPermitsthatthesampleexists. Destructionmusttakeplacewithinsevendaysofnotificationthatthesample ispositiveforR.solanacearumrace3biovar2andaPPQrepresentativemust havetheopportunitytowitnessthedestructionofthesampleorculture withinthattimeperiod.Theresponsiblelaboratorymanager/plant pathologistmustalsocompletetheAPHISForm2040.Theformisavailableat thePermitsWebsite.
Address PPQPermits Telephone: 301-734-7211, 6828 or 5055 Web site: http://www.aphis.usda.gov/permits/ppq_epermits.shtml
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5
Ralstonia solanacearum r 3 b 2
Chapter 5
Regulatory Procedures
Contents
Introduction page 51 InstructionsToOfficers page 51 OverviewOfRegulatoryProgramAfterAUnitedStatesDetection page 53 FurtherInvestigationsAtNurseriesWithSuspectGeraniums page 56 PlacingHolds page 57 RecordKeeping page 58 PlantstoHold page 59 IssuingPPQForm523EmergencyActionNotification page 511 GeraniumAndPotatoVolunteersAndCullPiles page 512 HarvestingEquipment,StorageAreas,AndConveyances page 512 RegulatoryRequirementsOfSeedProducers page 512 RegulatoryRequirementsForProcessors page 512 RegulatedArticles page 513 RemovingAreasFromQuarantine page 514 RequirementsForPotatoCroppingSystemsUnderQuarantine page 515 UseOfChemicals page 518
Introduction
UseChapter5RegulatoryProceduresasaguidetoproceduresthatmustbe followedwhensurveyingorcontrollingtheplantpathogenRalstonia solanacearumrace3biovar2.
Instructions To Officers
Agriculturalofficersmustfollowinstructionsforregulatorycontrolmeasures, treatmentsorotherprocedureswhenauthorizingthemovementofregulated articles.Afullunderstandingoftheinstructionsandproceduresisessential whenexplainingprocedurestopersonsinterestedinmovingarticlesaffected bythequarantineandregulations.Onlyauthorizedtreatmentsmaybeused inaccordancewithlabelingrestrictions.Duringallnurseryvisits,please assurethatpropersanitationproceduresarefollowed.SeeSurveyProcedures onpage 31andLawsPertainingtoPestControlonpage 61formore information.
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Afterinvestigationsareperformedandadditionalconsultationsmadeif necessarytheunitofheldsuspectgeraniumsandpotentiallyinfectedplants associatedwiththemmustbedestroyedordisposedof.Inaddition,thearea mustbedisinfected.SeeDisinfectionProceduresonpage 64formore information Ifapropertyhaspositivetestingorpositiveassociatedfields,allplant materialthatcanbereasonablyremovedfromthefieldmustbedestroyedor disposedof.Thisincludescullpilesandotherplantdebris. Leavethefieldfallowfortwoyears,andirrigatetopromotevolunteer sprouting.Duringthetwoyears,thevolunteercropmustbesampled,tested, anddiskedunderduringthegrowingseasoneveryfourweekstoeliminate hostmaterial.Weedhostsinthefieldandalongtheedgesmustbesprayed withefficaciouslabeledherbicidestoeliminatethemfromthearea. Inthefollowingtwoyears,plantfieldswithnonhostcrops;irrigateto promotevolunteerpotatosprouting,diskinganyvolunteerpotatoesas sproutingoccurs.Fieldsmustbetestedsemiannuallyforfouryearsafteran initialpositivefind.SeeSurveyProceduresonpage 31formore information. Furthermore,noseedproductioncanoccurinthefieldforatleastfiveyears afterdetectionofRalstoniasolanacearumrace3biovar2.Fieldswithsusceptible hosts(potatoesortomatoes)mustbesampledfortwoseasonspriortoany newseedproductionontheproperty. Fieldsadjacenttopositivetestingorassociatedfields,orthoseonashared watersupply,maynotgrowhostcropsfortwoyears,norseedpotatoesfor twoyears,andmusttestandcontrolanyvolunteerpotatoorweedhosts. Maintainstrictsanitationofallvehiclesenteringandleavingtheinfested fields. Storagefacilitiesonpropertieswithpositivetestingtubersmustbeheldin quarantineuntilallpotentiallyinfestedtubersaretestedandeitherfoundto benegativeordestroyed.Thestoragefacilitiesmustthenbeproperlycleaned anddisinfectedwithapproveddisinfectantsinstrictaccordancewithlabeling. SeeDisinfectantsonpage G1formoreinformation.
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Regulatory Records
Maintainstandardizedregulatoryrecordsanddatabase(s)insufficientdetail tocarryoutaneffective,efficient,andresponsibleregulatoryprogram.
Conductatracebackinvestigationtodeterminetheforeignsourceofinfection andtheextentofdistributionofpotentiallyinfectedplantsintheUnited Statesfromthatsource.Conductatraceforwardinvestigationtodetermineif potentiallyinfectedplantmaterialhadbeendistributedtootherretail locations. InvestigationsatUnitedStatesgeraniumpropagationfacilitiesandtheir customerswillbeconducted.Resultsofinvestigationswillallow implementionofaprogramofholdingvarietiesorshipmentsthatoriginated ataparticularforeignfacilityduringashippingperiod.Theirrigation methods,handling,andsanitationpracticesofeachnurserywillhelpassessif additionalassociatedplantmaterialisatriskofinfection.Information
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Conductatracebackinvestigationtodeterminethesourceofinfectedtubers andtheextentofdistributionofseedtubersintheUnitedStatesfromthat source.Conductatraceforwardinvestigationtodetermineifpotentially infectedtubershavebeendistributedtogrowersorprocessingplantswhere processedmaterialandwatermaybereleasedtotheenvironment.See TracebackAndTraceForwardSurveyonpage 37formoreinformation. Informationgatheredintheinvestigationwillbecriticaltodeterminingwhich additionaltuberlotsandpropertiestohold,aswellaswhichactionstotake. Consultwithregionalcontactsifguidanceisneeded.
Alistofnurseryfacilitieswillbecompiledofgeraniumvarietiesand/or shippingwindowsthathavebeen associated with positively tested, foreign origin geraniums. Theselistswillbedistributedbystatetothefieldoffices,andare nottobesharedwithindividualsoutsideourregulatorycooperators. Companynamesandlocationsontheselistsarestrictlyconfidential,andany distributionoflistsbeyondappropriateregulatoryagencycontactsis prohibited. Whennotifyingnurseriesonthelist,besuretoidentifyyourselfasaUSDAor stateregulatoryofficialconductinganinvestigationofnurseriesthatmay havereceivedRalstoniasolanacearuminfectedmaterial.Speaktothenursery ownerormanager. Itisimportanttounderstandthatnurseryfacilitieshandlingforeign geraniumsareofseveraltypes.Companiesshipcuttingsandcallusedplants throughPPQPlantInspectionstations.Theclearedmaterialistheneither senttorootingstationsortodirectshipcustomerswhicharewholesalers.
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Bothtypesoffacilitiesgrowtheplantsforaperiodoftime(usually5weeks) beforebeingdistributedtootherentities.Rootingstations,undercontract withthecompany,thenshiporderstoavarietyofcustomertypesincluding wholesalersandbothlargeandsmallretailcustomers.Directshipfacilities canhavetheirownnetworkofretailstoresorsellplantstoothercustomers. Handlingandculturalpracticesvaryatallstepsinthedistribution. Duringtheinvestigation,requestthatthenurseryownerormanagerproduce anytags,invoices,shippinglistsassociatedwiththepositivetestedsuspect plantsonthedistributedlist.Makecopiesoftheserecordsandlocatethe suspectplantsstillonthepremises.Takephotographsoftheareawhere suspectgeraniumsarelocatedandanygreenhouseconditionsincludingthe proximityofneighboringplantsthatmayindicatebroadercontamination. Makearecordofthenumberofplants,type,andlocationincludingan accountingofsuspectplantsfromthatshipmentnotonthepremises. Tracebackinformationgatheredfromplanttagsandinvoicescanbeusedto determinetheoriginoftheplants.Withtimelysubmittedrecordsfromplant distributioncompaniessuchasrootingstationsorwholesalegrowers, PPQEmergencyandDomesticProgramsstaffcangeneratelistsofnurseries withimplicatedplantvarieties.Considerableeffortshavebeenmadebythe geraniumindustrytousebarcodingforplantsandcuttingstrackingthatmay haveuseintheseinvestigations. Thisinformationcanbeusedtodeterminetheextentofdistributionofsuspect geraniumsatvariouslocationsintheUnitedStates.Coordinatewithyour statecontactsandregionalofficetogatherthisinformation.Additional resourcesmaybeneededtoquicklygatherthisinformationandanalyzeitin ordertoholdsuspectgeraniumshipmentsandpotentiallyinfectedplants.
Potato Cropping Systems
storage,atshippingpoints,orinprocessingfacilitiesthatareconfirmed positiveforRalstoniasolanacearumrace3biovar2.
Positive AssociatedSeedorwarepotatoes,infields,orasharvested,orin
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Placing Holds
Nurseries
Afteradestinationnurseryisidentifiedashavingreceivedplantsfrom anothersourceassociatedwithpositivetestinggeraniums,itisimportantto immediatelyplaceholdsonallgeraniumvarietiesnamedondistributedlists andinitiallyanyotherhostsplantsinthefacility.Iftheextentofpotentially infectedplantsatthatfacilitycanbeascertainedatthattime,thosenot determinedtobepotentiallycontaminatedcanbeimmediatelyreleased. Aholdisinterpretedastheprohibitionofmovementofthoseplantsfromthe propertyuntilfurtherevaluationscanbemadeandcontrolactionscanbe takenonsuspectgeraniumsandotherpotentiallyinfectedplants. Afterinvestigationsaremade,unlessgeneralcontaminationissuspected,the holdisnottobeinterpretedto: Prohibitthemovementofallplantsinagreenhouse; Thatplantsnotheldcannotbecaredforinanormalandsanitary manner;and Prohibittherelocationofsuspectgeraniumshipmentsandpotentially infectedplantstoasegregatedareaawayfromotherplants;ifthe nurseryowner/managerwishestomoveheldplants,assesstheriskof suchmovementandkeeprecordsofwhererelocatedplantsaremoved.
Ralstonia solanacearum r 3 b 2
5-7
Record Keeping
Geraniums
Recordkeepinganddocumentationisimportantforanyholdsand subsequentactionstaken.Relyonshippingrecordsandinformationprovided bythenurseryowner/managerforhowmanyplantsremain,howplantshave movedwithinthenursery,destinationofplantssold,andculturalpractices employed. Keepadetailedaccountingofthenumbersandtypesofeachplantvariety heldand/ordestroyedincontrolactions.Consultamasterlistofvarieties distributedwiththelistsoffacilities.Drawmapsofthegreenhouselayoutto locatedsuspectgeraniums,otherpotentiallyinfectedplants,andwaterrunoff areasincludingrecirculatingpondlocations.Takephotographsofthefacility layout,geraniumplacement,wateringmethod,materialsandmethodsused, plantlabeling,andanyothersituationsthatmaybeusefulfordocumentation andanalysis.
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Ralstonia solanacearum r 3 b 2
Plants to Hold
CompletePPQForm523EmergencyActionNotificationonpage B7to placeholdsatdestinationnurseriesforthefollowingclassesofplants:
Suspect Geraniums
Variety
Dependingontheparticularprogramdirections,aparticularvarietyor varietiesofgeraniumplantsmaybeconsideredsuspectbecausetheytested positiveandtracebackinvestigationstoaparticularforeignfacility determinedtheiroverallinfectionrisk.Thevarietylevelholdcanbemost appropriateatrootingstationsordirectshippedgrowersthatarelarge operationswhosepracticeistokeepvarietiesseparate. Dependingonprogramdirectionsandpracticesatapreviousfacilityinthe distribution,alevelofsuspectgeraniumstoholdmayincludetheshipmentof allhostplantsreceivedfromthatfacility. Releaseallplantsthatarenotconsideredpotentiallyinfected.Thefollowing categoriesaresubjecttohold.
Shipment
High RiskResultsofinvestigationsatnurseryfacilities,andanswerstothe
questionnaire,willdeterminethelevelofadditionalholdsofpotentially infectedplants.Afterinvestigationsaremade,someoftheseconditionswill requireindividualjudgmentandconsultationwithyourregionaloffice.The categoriesthatdefinehigherriskforpotentiallyinfectingplantsinclude: PlantsbeneaththedriplineofsuspectgeraniumsBecauseRalstonia solanacearumbacteriaareeasilyshedinwater,plantsdirectlybelow hanginggeraniums,regardlessoftheplantspecies,areathighriskof soilcontaminationandmustbeincluded.Thisincludeshostsand nonhosts.Thisisalsotrueforplantsstoredunderbencheswithsuspect geraniums. PlantsinthesamepotswithsuspectgeraniumsInfectedgeraniumsin thesamepotswithotherplantscaneasilyinfectthemthroughwater, soil,orrootcontact.Thisincludeshostsandnonhosts. PlantpropagationfromsuspectgeraniumshasoccurredAnyplants propagatedfromsuspectgeraniumsareathighriskforinfection. PlantsonasharedirrigationsystemwithsuspectgeraniumsThemost efficientmethodofspreadforR.solanacearumisthroughwaterthat cameincontactwithinfectedplants.Itisthereforenecessarytoholdall
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5-9
plants(hostsandnonhosts)thatareonirrigationsystemsthatallows forwaterflowfromoneplanttothenext.Thesearevariousirrigation situationstolookfor: Subirrigation,ebbandflow,orfloodirrigation:potssitinapanand areirrigatedbyfloodingthepaninvariousways. Backflowprevention:nearlyallirrigationsystemshavecheckvalves toasasafeguardtopreventcontaminatedwaterfrombackingup intothegeneralwatersupply.Ifsystemslackbackflowprevention, thereisahighriskofgeneralcontaminationthroughoutthesystem frominfectedmaterial. Plantsinfacilitieswheresanitaryculturalpracticesarenotinplace Usinggraftingknivesformakingcuttingsorgroomingplantswithout disinfectionbetweenvarietiesisahighriskfactorintransmittingthe pathogentootherhostplantlots. Plantsplacedontheground,onplasticsheetingorothermaterialthat allowspuddlingbetweenplantsBecausethepathogenspreadsby water,thereisariskthatpuddlingbetweenplantswillcauseuptakeof bacteriabyhealthplantsinthevicinityofinfected.Theextenttowhich thisoccursneedstobeassessedbyobservationandfurtherevaluations ormonitoringmaybenecessary.
Reduced RiskThefollowinggreenhouseculturalpracticesareconsidered
notashighofriskforspreadingthepathogentootherplants.Evaluationscan bemadebytheinspectorastothedegreetowhichsuchpracticesmaytake placebasedonobservationsandresponsestothequestionnaire.Itis recommendedthattheinspectorconsultwiththesupervisor,SPHD,SPROor regionalofficeifthereisreasontobelievetheextentofthesepracticespresents agreaterriskandthereforerequiresadditionaldestructionofplants.Insome cases,afterconsultation,thedegreetowhichtheseriskfactorsarepracticedat thefacilitymaydeterminetheneedforwiderdestructionofpotentially infectedplants,however,inmostcases,thesenurserieswithplantsinthese reducedriskcategoriesmaybeheldandsubjecttofollowupinspectionsto inspectforwiltsymptoms. PlantsthatwerenotsegregatedfromsuspectgeraniumsRecent informationshowsthatleaftoleafcontactbetweengeraniumsisnotan effectivemethodofpathogenspread,sothereislittletonoriskofplants inthesameproximityofinfectedplantsbecomingcontaminatedby contact.Anexceptiontothismightbeplantsimmediatelyadjacentto infectedplantsthatwerewateredbyhand(usingahoseora waterwand)wheresplashmaytransferthepathogentothesoilin adjacentplants.Generally,dripirrigationandmistsystemsdonot createariskofsplashtoadjacentplants.Whenindoubt,consultyour SPHD,SPRO,andregionalcontact. Plantsthatwerepinched,deleafed,disbuddedbyhandThismethod ofgroomingpresentslessriskofspreadtononinfectedhostsplants thanbytheuseofcuttingtools,butsomedegreeofsanitationbetween
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received,thesameEANforwhichtheplantsareonholdisusedtodocument anyactionstakensuchasdestructionanddisinfection.Ifvarietiesor shipmentsaretobeheldasseparateunits,itisadvisabletoissueseparate EANsforeachheldunitofsuspectgeraniumsorpotatoesandpotentially infectedplantsassociatedwiththatunit. ElectronicEANscanbeused.SeePPQForm523EmergencyAction Notificationonpage B7forinstructions.EANsareissuedunderthe authoritygivenin7CFR330.Statesshouldissuetheirownholdorders paralleltotheEANtoassureplantscannotmoveintrastate.
Address http://www.aphis.usda.gov/library/forms/
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Ralstonia solanacearum r 3 b 2
receivedpositivetestingorpositiveassociatedstock,oriftestsarepositive, thewaterandequipmentusedinprocessingandthewasteproductsfrom processingwillrequireregulatoryaction.Placeholdsonfacilities,water sources,highriskplantmaterialfromprocessinguntilassessmentsaremade todeterminewhichpartsofthefacilitywillrequiredisinfection,water treatment,andproperdisposalofhighriskwastematerial. Processorreceivingpositivetestingorpositiveassociatedpotatoesmust destroyordisposeofallofthosepotatoesintheirinventory.Allstorageareas, processingequipment,andconveyancesmustbedisinfected.Allwater sourcesorstoragepondsrecirculatedforprocessingmustbetestedand decontaminatediffoundtobepositive.
Regulated Articles
Cultivated Host Plants
Plants,cuttings,orpartsofthefollowinghostsofRalstoniasolanacearumrace3 biovar2areregulatediftestedpositiveandconsideredpotentiallyinfected.
TABLE 5-1 Cultivated Hosts of Ralstonia solanacearum race 3 biovar 2
Geranium Tomato Peppers Eggplant Potato Bean Bittergourd Beet
1
Pelargonium spp.1 Lycopersicon esculentum Capsicum spp. Solanum melongena Solanum tuberosum Phaseolus vulgaris Momoridica charantia Beta vulgaris
Ralstonia solanacearum r 3 b 2
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5-14
Ralstonia solanacearum r 3 b 2
Storage Facilities
Storagefacilitiesonpropertieswithpositivetestingtubersmustbeheldin quarantineuntilallpotentiallyinfestedtubersaretestedandeitherfoundto benegativeordestroyed.Thestoragefacilitiesmustthenbeproperlycleaned anddisinfectedwithapproveddisinfectantsinstrictaccordancewithlabeling. SeeDisinfectantsonpage G1formoreinformation.
Ralstonia solanacearum r 3 b 2
5-15
Then: Destroy or dispose of all plant material that can be reasonably removed from the field, including: Cull piles Other plant debris Storage facilities on the properties must be held in quarantine until all potentially infested tubers are tested and either found to be negative or destroyed. The storage facilities must then be properly cleaned and disinfected with approved disinfectants in strict accordance with labeling. See Disinfectants on page G-1 for more information.
Fields are adjacent to positive testing or associated fields, or are on a shared water situation
Do not grow host crops or seed potatoes, Test and control any volunteer potato or weed hosts. Maintain strict sanitation of all vehicles entering and leaving the infested fields.
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Ralstonia solanacearum r 3 b 2
5-17
Use Of Chemicals
ThePPQTreatmentManualandthisGuidelinesidentifytheappropriately labeledchemicals,anddescribethemethodsandratesofapplication. ConcurrencebyPPQisnecessarybeforeusinganyotherchemicalor procedureforregulatorypurposes.
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6
Ralstonia solanacearum r 3 b 2
Chapter 6
Control
Contents
Introduction page 61 HandlingOfSamples page 62 PlantDisposalAndDestructionMethods DisinfectionProcedures page 64 ApprovedDisinfectants page 66 page 63
Introduction
UseChapter6ControlasaguidetothecontrolofRalstoniasolanacearumrace3 biovar2.USDAAPHISPlantProtectionandQuarantine(PPQ)developsand makescontrolmeasuresforR.solanacearumrace3biovar2availableto involvedstates.EnvironmentalProtectionAgency(EPA)approvedand labeledtreatmentswillberecommendedwhenavailable.
1
May 12, 2008 Emergency and Domestic Programs Control Procedures.fm Ralstonia solanacearum r 3 b 2 6-1
Section24Astatecanprovideregistrationforadditionalusesof federallyregisteredpesticidesformulatedfordistributionanduse withinthatstatetomeetspeciallocalneedsinaccordancewiththe purposesofthisact. Foradditionalinformationconcerningexemptions,seetheEmergency ProgramsManual,Section14.ContactEnvironmentalServicesstafftoassure thatanypesticidebeingconsideredaspartofaneradicationprogram conformstopesticideuserequirements.Obtainallrequiredenvironmental documentationbeforebeginning.SeeEnvironmentalComplianceon page A1formoreinformation.
Control Records
Programpersonnelmustmaintainrecordsandmaps,notingthelocationsof alldetections,thenumberandtypeofplantssubjectedtocontrolactions,and thematerialsandformulationsusedineachtreatedarea.Attachall documentationandreceiptstotheofficecopyofthePPQForm523 EmergencyActionNotification.SeePPQForm523EmergencyAction Notificationonpage B7formoreinformation.
Handling Of Samples
Placeallpotentiallyinfectedplantsindoubleplasticbagsandsealfordisposal ordestruction,includingthefollowing: Shipmentsofsuspectgeraniums AssociatedpotatoplantsortubersthathavetestedpositiveforRalstonia solanacearumrace3biovar2 Pots,directlyassociatedsoilorpottingmedia,andanyotherdisposable itemincontactwiththeplantsorsoil Hostweedsinpositivetestedfacilitiesmustalsoberemovedandcorrectly disposedofordestroyed.SeePlantDisposalAndDestructionMethodson page 63formoreinformation Maintainanaccuratecountofeachtype(variety,etc.)ofplantortubers baggedfordestructionandrecordthisinformation.
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Iflargeinventoriesmustbedestroyed,theuseofplasticbagsmaynotbe reasonable.Alternativesmayincludeadumpsterwithdoublelayersofplastic liningthatcanbefoldedoverthetopandsealedtopreventdebrisfrom escapingduringtransportorstorage.ContactyourregionalofficeorPPQ Headquartersforadditionalguidance. Allbenchareaswhereplantswereheld,andotherareasatriskforexposureto infectedplantmaterial,shouldbedisinfectedaccordingtoprescribed disinfectionprocedures.SeeDisinfectantsonpage G1formore information.
Burning or Incineration
Allplantmaterialandmediamustbeincineratedorburnedtothepointof ash.Ifplasticpotsarenotacceptedattheincinerator,removethemand properlydisinfectorsendtoalandfill.
Soil Fumigation
Soilfumigationwithanapprovedfumigantsuchasmethylbromideor metamsodiumatthelabeledratewillgreatlyreducepopulationsofthe bacteriaininfestedfields.However,fieldswithhostplantresidueincluding tubers,roots,andstems,maystillharboradequateamountsofRalstonia solanacearumforuptotwoyears.Beforefumigation,removeanddestroyas muchhostplantmaterialfromthefieldsaspossible. Fumigationofthesoilmaynottotallyeliminatethepathogenduetothe presenceofbacterialreservoirsinburiedplanthostresidueatthetimeof fumigation.ContinuedmonitoringofthesoilforthepresenceofRalstonia solanacearumrace3biovar2willberequired.
Approved Landfill
Anapprovedlandfillisastatelicensedmunicipalorprivatefacilitythatis managedunderstateregulationtomeetconditionsthatwouldprevent potentialpollutantsfromleachingintogroundwater.
Ralstonia solanacearum r 3 b 2
6-3
Theregulatoryofficialthatwitnessesthedisposalofdoublebaggedmaterial fromnurseriesorpotatocroppingsystemsmustassurethematerialisburied undertwoormorefeetofsoil.Notifythelandfillinadvancetoarrangefora holetobeduginwhichthematerialcanbedisposedandcoveredwithsoil.If diggingaholeisimpracticalforalandfilloperation,anotheroptionistocover theplantmaterialwithalternativedailycoverorcompostedgreenwaste normallyusedtocoverrefuseatlandfills.Theseoperationsmustbe witnessed,supervised,anddocumentedbyaplantregulatoryofficial. Thequantitiesofpotatohostmaterialorderedfordestructionmaybe impracticalforthedisposaloptionsprescribed.Compostingwilllikelyheat thetemperatureofhostmaterialtosufficienttemperaturesnecessarytokill thepathogen.Theparametersofacomposingdisposalmethodmustbe proposedbyaRalstoniatechnicalgroup.Theefficacyofcompostingmustbe testedbyCPHSTpriortoimplementationasaprogrammethod.Caremustbe takentocontrolwaterrunofffromcomposingoperationsifimplemented.
Disinfection Procedures
Usethefollowingprocedurestodisinfectallpotentiallycontaminated surfaces(benches,flats,walkways,footbaths,drainageareasunderbenches) andequipmentwithinaninfectedgreenhouseorareaincontactwithinfected material: 1. Cleanawayanysoilormedia. 2. Sprayandsoaktothepointofrunoffwithapproveddisinfectantsorby pressurizedsteamcleaningtoraisethesurfacetemperatureto212oF. Formoreinformation,consultthefollowingsources: ApprovedDisinfectantsonpage 66 Disinfectantsonpage G1 PPQTreatmentManual
6-4
Ralstonia solanacearum r 3 b 2
Holding Ponds
Whenholdingpondshavebecomecontaminatedfromirrigationsystem runoff,contactyourregionalPPQofficeandStateaboutpossible environmentalconsiderationsandtreatmentoptions.
Ralstonia solanacearum r 3 b 2
6-5
Approved Disinfectants
UsetheapproveddisinfectantsinTable 61todisinfectalltools,benches, walkways,surfaces,andgravelbedsunderbenches,incontactwith potentiallyinfectedplantmaterialafterremovalofsoilormedia.Besurethat thelabelspecifiesRalstoniaorPseudomonas.CheckwithStatelabeling requirements.SeeDisinfectantsonpage G1formoreinformation
TABLE 6-1 Approved Disinfectants
Product Quaternary Ammonia Physan 20 Green Shield Maquat 615-HD or 615-LR ZeroTol
Notes Several quaternary ammonia (20% ammonium chloride) products are approved for greenhouse use. Physan 20 is not approved for greenhouse where food crops are grown. It is not approved for use in California Green Shield is approved for most uses. In California, use the special formulation Green ShieldCA. None ZeroTol is 27% hydrogen dioxide. It is not a quaternary ammonia, and has rates for disinfection including surfaces with soil or media contamination that cannot be cleaned. This product is also approved for use in disinfection of contaminated irrigation systems.
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7
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Chapter 7
Environmental Compliance
Contents
Introduction page 71 Disclaimer page 71 NationalEnvironmentalPolicyAct page 72 EndangeredSpeciesAct page 72 FederalInsecticide,fungicide,AndrodenticideAct OtherLaws page 73 EnvironmentalMonitoring page 73
page 73
Introduction
UseChapter7EnvironmentalComplianceasaguidetoenvironmental regulationspertinenttotheplantpathogenRalstoniasolanacearumrace3 biovar2ingeraniums,potatoes,andothersolanaceoushosts. Akeyelementindesigningaprogramoranemergencyresponseis consultationwithEnvironmentalServices(ES),aunitofAPHISPolicyand ProgramDevelopmentstaff.ESperformsthefollowingfunctions: Preparesenvironmentaldocumentationsuchasenvironmentalimpact statementsandenvironmentalassessmentstoaidinmakingdecisions Providesconsultationonthetopicofendangeredspecies CoordinatespesticideregistrationandapprovalsforAPHISprograms, ensuringthatregistrationsandapprovalsmeettheneedsofprograms andconformtopesticideuserequirements Inaddition,PPQsEnvironmentalComplianceteamassistsESinthe developmentofrequireddocumentationandimplementsanyenvironmental monitoringthatmayberequiredofprogramactivities.RefertoResourceson page A1foradditionalinformation.
Disclaimer
AllusesofpesticidesmustberegisteredorapprovedbyappropriateFederal, State,and/orTribalagenciesbeforetheycanbeapplied.Theinformation providedonpesticidelabelsmaynotreflectalloftheactualinformation, includingprecautionsandinstructionsforuse,whichyouarerequiredto followinyourspecificstateorlocality.Itistheresponsibilityofpersons
May 12, 2008 Emergency and Domestic Programs Env.fm Ralstonia solanacearum r 3 b 2
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7-2
Ralstonia solanacearum r 3 b 2
Other Laws
TheNationalEnvironmentalPolicyAct,EndangeredSpeciesAct,andthe FederalInsecticide,Fungicide,andRodenticideAct,areofcriticalimportance toallpestcontrolprograms,butotherlawsmayapplydependingonprogram locationsandactivities.TheseincludetheMigratoryBirdTreatyAct,the CoastalZoneManagementAct,andtheBaldandGoldenEagleProtection Act.ByincludingEnvironmentalServicesandEnvironmentalCompliance earlyinprogramplanning,guidancecanbeprovidedonmeetingthe requirementsoftheseandotherlawsthatmayapply.RefertoResourceson page A1forcontactinformation.
Environmental Monitoring
EnvironmentalmonitoringofAPHISpestcontrolactivitiesmayberequired aspartofcompliancewiththeabovelaws,asrequestedbyprogram managers,orassuggestedtoaddressconcernswithcontroversialactivities. Thisisespeciallytrueforlessbenignchemicalcontrolsandaerialapplication ofchemicals. Monitoringmaybeconductedwithregardstoworkerexposure,quality assuranceandcontrol,offsitedeposition,orprogramefficacy.Differenttools andtechniquesareuseddependingonthemonitoringgoals,program chemicals,andcontroltechniques.Environmentalmonitoringiscoordinated byEnvironmentalCompliance(EC).StafffromECwillworkwiththe programmanagertodevelopanenvironmentalmonitoringplan,conduct trainingtoimplementtheplan,providedaytodayguidanceonmonitoring, andprovideaninterpretivereportofmonitoringactivities.
Ralstonia solanacearum r 3 b 2
7-3
7-4
Ralstonia solanacearum r 3 b 2
Ralstonia solanacearum r 3 b 2
Glossary
approvedlandfill.Statelicensedmunicipalorprivatelandfillthatismanaged understateregulationtomeetconditionsthatwouldpreventleachingof potentialpollutantsintogroundwater. biovar.Groupofbacterialstrainsthataredistinguishablefromotherstrainsof thesamespeciesonthebasisoftheirphysiologicalcharacteristics. chlorosis.Yellowingofnormallygreentissueduetochlorophylldestruction ininfectedplants. decontamination.Applicationofanapprovedchemicalorothertreatmentto contaminatedimplements,material,orbuildingsforkillingordeactivatinga pathogen. detectionsurvey.Surveyconductedinanenvironmentallyfavorablearea whereRalstoniasolanacearumrace3biovar2isnotknowntooccur. destinationnursery.Anynurseryreceivingsuspectgeraniumsorsuspect plantsfromarootingstationorothernursery. disposal.Methodusedtoeliminatediseasedplantmaterialormaterial associatedwithdiseasedplantmaterial,usuallyatanapprovedlandfill. ELISA.AcronymforEnzymeLinkedImmunosorbentAssay,aserological laboratorytechniqueusedtodeterminethegenusandspeciesinaRalstonia hostsamplingandtestingprogram;excludesraceandbiovar. EnzymeLinkedImmunosorbentAssay.SeeELISAabove. host.Plantwhichisinvadedbyaparasiteorpathogenandfromwhichit obtainsitsnutrients. incineration.Burningofplantsandassociatedsoilormediathatresultsin theircompletedestruction. infection.Establishmentofaparasiteonorwithinahostplant. monitoringsurvey.Surveyconductedatasitewhereadiseasewasfoundand whereaneradicationprogramisbeingperformed. necrosis.Deadordiscoloredplanttissue.
1
Glossary-1
Ralstonia solanacearum r 3 b 2
pathogen.Microorganismthatcaninciteadisease. PCR.AcronymforPolymeraseChainReaction,alaboratorytechniquethat amplifiesDNAsequencesinordertodeterminetheraceandbiovarina Ralstoniahostsamplingandtestingprogram. positiveassociated.Seedorwarepotatoes,infields,orasharvested,orin storage,atshippingpoints,orinprocessingfacilitiesthataredirectlylinkedto positivetestingforRalstoniasolanacearumrace3biovar2. positivetesting.Seedorwarepotatoes,infields,orasharvested,orin storage,atshippingpoints,orinprocessingfacilitiesthatareconfirmed positiveforRalstoniasolanacearumrace3biovar2. potentiallyinfestedtubers.Seedorwarepotatoes,infields,orasharvested, orinstorage,atshippingpoints,orinprocessingfacilitiesthatareindirectly linkedtopositivetesting,forRalstoniasolanacearumrace3biovar2. PolymeraseChainReaction.SeePCRabove. potentiallyinfectedplants.Withinthenurseryproperty,theseinclude: Allplantsunderthedriplineofhangingsuspectgeraniums Otherplantsthathavebeenplantedinthesamepotswithsuspect geraniums Allplantsonasharedwaterirrigationsystemwithsuspectgeraniums, includingthefollowing: Ebbandflow Flood Subirrigation Systemsthatlackofbackflowprevention Allhostplantsthatmayhavebeeninfectedbyapositivetestingsuspect geraniumshipmentthroughunsanitarygreenhousepractices,including thefollowing: Failuretodisinfecttools,hands,orequipmentduringgrafting, pruning,debudding Deleafingbetweenvarieties Plantsonthegroundwithanonporoussurfacesuchthat puddlingmayoccurunderplants
Glossary-2
Ralstonia solanacearum r 3 b 2
potentiallyinfectedplantswithinnurseryproperty.Includesallplantsthat arelocatedunderthedriplineofhangingsuspectgeraniumsoronashared irrigationsystemwithapositivetestingsuspectgeranium.Alsoincludes plantsthatmayhavebeeninfectedbypositivetestingsuspectgeraniums throughunsanitarygreenhousepractices. positivetestinggeraniums.GeraniumsinwhichRalstoniasolanacearumrace3 biovar2hasbeenconfirmedtobepresentbytheUSDAPPQCPHST laboratoryinBeltsville,MD. shipment.Allplantsthatwerepreparedforshipmentandtransportedwith suspectgeraniumsfromacommonsupplier,i.e.,listedontheinvoiceor packinglist. suspecthosts(geraniumsorpotatoes).Geraniums(Pelargoniumspp.)thatare associatedwithpositivetestinggeraniums.Thesemayhaveoriginatedfroma foreigncountry,arootingstation,oranothernurseryandbethesamevariety orinthesameshipmentdependingonthecircumstancessuspecttubers.Also, potatotubersinfieldsorstoragefacilitiesthatareassociatedwithpotato tubersthathavetestedpositiveforRalstoniasolanacearumrace3biovar2. suspectgeraniums.Geraniums(Pelargoniumspp.)thatareassociatedwith positivetestinggeraniums.Thesemaybeaparticularimplicatedvarietyor shipmentthatoriginatedfromarootingstation,directshipfacility,oranother nursery. suspectshipment.Allhostplantsthatwerepreparedforshipmentand transportedwithsuspectgeraniumsfromacommonsupplier(listedonthe invoiceorpackinglist). symptom.Theexternalandinternalreactionsoralterationsofaplantasthe resultofadisease. traceback.Toinvestigatetheoriginofinfestedplantsthroughintermediate stepsincommercialdistributionchannelstotheorigin. traceforward.Toinvestigatewhereinfectedplantsmayhavebeendistributed fromasourcethroughstepsincommercialdistributionchannels.
Ralstonia solanacearum r 3 b 2
Glossary-3
Glossary-4
Ralstonia solanacearum r 3 b 2
Ralstonia solanacearum r 3 b 2
References
Buddenhagen,I.W.,L.SequeiraandA.Kelman,A.1962.Designationofraces ofPseudomonassolanacearum.Phytopathology52:726. CABI.2003.CropProtectionCompendium.UnitedKingdom:Wallingford: CABInternational.Onlinedatabase.http://www.cabicompendium.org/cpc. AccessedNovember4,2003. CABI.2006.CropProtectionCompendium.UnitedKingdom:Wallingford: CABInternational.Onlinedatabase.http://www.cabicompendium.org/cpc. AccessedJuly2007. Cook,D.R.andL.Sequeira.1994.StrainDifferentiationofPseudomonas solanacearumbyMolecularGeneticMethods.,pages7793.In:A.C.Hayward andG.L.Hartman(editors).BacterialWilt:TheDiseaseanditsCausative Agent,Pseudomonassolanacearum.CABInternational,Wallingford,UK.259pp. Daughtrey,M.March4,2003.SouthernBacterialWilt,causedbyRalstonia solanacearum.In:SummaryofPresentationatSocietyofAmericanFlorists19th AnnualConferenceonInsectandDiseaseManagementonOrnamentals. http://www.greenhouse.cornell.edu/pestsdis/MD.SAFtalk.pdf. DepartmentforEnvironment,FoodandRuralAffairsofUnitedKingdom (DEFRA).2003a.PlantPathology.www.defra.gov.uk/planth/phnews/ openday/brown.pdf.AccessedMarch2003. DepartmentforEnvironment,FoodandRuralAffairsofUnitedKingdom (DEFRA).2003b.PlantPathology.http://www.defra.gov.uk/planth/stake.pdf. AccessedMarch2003. DepartmentforEnvironment,FoodandRuralAffairsofUnitedKingdom (DEFRA).2003c.PlantPathology.http://www.defra.gov.uk/science/LINK/ Publications/Newsletters/AgricultureLINK/AgLINK_Issue3.pdf.Accessed March2003. Denny,T.P.andA.C.Hayward.2001.Ralstonia,pages151174.In:Schaad,N. W.etal.LaboratoryGuidefortheIdentificationofPlantPathogenicBacteria, 3rded.APSPress.Minnesota:St.Paul.373pp. Douglas,S.M.2002.DiseasesofGeranium.TheConnecticutAgricultural ExperimentStation.Connecticut:NewHaven.
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EconomicResearchService(ERS).2003.StatisticalIndicators. USDAEconomicResearchServicePublications.http://www.ers.usda.gov/ publications/. French,E.R.,L.Gutarra,P.AleyandJ.Elphinstone.1995.Culturemediafor Pseudomonassolanacearum:isolation,identificationandmaintenance. Phytopathologia30(3):126130. FrenchE.R.1998.RecentSpreadofBacterialWiltofPotato:Regional PerspectivesandExperiences,LatinAmerica,pages426427.In:Prior,P.,C. AllenandJ.Elphinstone(eds.).BacterialWiltDisease:Molecularand EcologicalAspects.ReportsoftheSecondInternationalBacterialWilt SymposiumheldinGosier,Guadeloupe,France,2227June,1997. SpringerVerlag.BerlinHeidelberg,INRA,Paris.447pp. Garret,L.J.2003(Draft).PotentialLossesofRalstoniasolanacearumrace3, biovar2.USDAAPHISCPHST,Raleigh,NC. Hayward,A.C.1964.CharacteristicsofPseudomonassolanacearum.J.Appl. Bacteriol.2:265277. Hayward,A.C.1991.Biologyandepidemiologyofbacterialwiltcausedby Pseudomonassolanacearum.Annul.Rev.Phytopathol.29:6587. Horita,M.andK.Tsuchiya.2001.GeneticdiversityofJapanesestrainsof Ralstoniasolanacearum.Phytopathology91:399407. Hudelson,B.D.May11,1999.SouthernBacterialWilt.Univ.Wisconsin GardenFacts. Janse,J.D.1991.InfraandintraspecificclassificationofPseudomonas solanacearumstrainsusingwholecellfattyacidanalysis.SystematicandApplied Microbiology14:335345. Janse,J.D.1996.PotatoBrownRotinWesternEuropeHistory,Present OccurrenceandSomeRemarksonPossibleOrigin,EpidemiologyandControl Strategies.Bull.OEPP26:679695. Janse,J.D.,H.E.VanDenBeld,J.Elphinstone,S.Simpkins,N.TijoiTamSin, andV.Vaerenbergh.2003.IntroductiontoEuropeofRalstoniasolanacearum Biovar2/Race3inPelargoniumzonaleCuttings.AmericanPhytopathological Society. Kelman,A.1953.ThebacterialwiltcausedbyPseudomonassolanacearum.A literaturereviewandbibliography.NorthCarolinaAgriculturalExperiment StationTechnicalBulletinNo.99.194pp. Kim,S.H.andT.N.Olson.2003.RalstoniasolanacearumRace3,Biovar2,the CausalAgentofBrownRotofPotato,IdentifiedinPennsylvania,Delaware andConnecticut.PlantDisease(87)4:450.
References-2
Ralstonia solanacearum r 3 b 2
Lee,et.al.2001.PlantDisease85:261266. Meiners,U.2000.Bacterialringrot:possibilitiesandlimitsofinsurance coverage.http://ibws.nexenservices.com/from_july_%2001.htm. Melton,T.A.,andH.D.Shew.2000.GranvilleWilt.TobaccoDisease InformationNote.TDIN002.NorthCarolinaStateUniversity,Collegeof AgricultureandLifeSciences,PlantPathologyExtension.http:// www.ces.ncsu.edu/depts/pp/notes/Tobacco/tdin002/tdin002.htm. NorthAmericanPlantProtectionOrganisation(NAPPO).2001.Ralstonia (Pseudomonas)solanacearum(E.F.Smith,1896)Yabuuchietal.1995race3 biovar2.PhytosanitaryAlertSystem.http://www.pestalert.org.Accessed March2003. NationalInstituteofAgricultureScienceandTechnology.2003.Research Highlights.http://www.niast.go.kr/english/sub/highlights.htm.AccessedMay 15,2002. Momol,T.,J.Jones,andS.Olson.2003.NewOutbreakofRalstonia solanacearum(race3biovar2)ingeraniumsinUnitedStatesandeffectsof biofumigantsonRalstoniasolanacearum(race1biovar1).UniversityofFlorida PestAlert.http://extlab7.entnem.ufl.edu/PestAlert/tmm0303.htm. Pittman,H.A.1933.BacterialWiltofTomatoesandotherSolanaceousCrops. West.Austral.Dept.Agr.J.(II)10:373374. PlantProtectionandQuarantine(PPQ).2004.MinimumSanitationProtocols ForOffshoreGeraniumCuttingProduction.USDAAPHISPPQ.http:// www.aphis.usda.gov/plant_health/plant_pest_info/ralstonia/downloads/ ralstoniaworkplan.pdf.AccessedAugust20,2007. Prior,P.andH.Steva.1990.CharacteristicsofstrainsofPseudomonas solanacearumfromtheFrenchWestIndies.PlantDisease74(1):1317. Possier,S.,P.VandewalleandJ.Luisetti.1999.GeneticdiversityofAfricanand worldwidestrainsofRalstoniasolanacearumasdeterminedbyPCRrestriction fragmentlengthpolymorphismanalysisofthehrpgeneregion.Appliedand EnvironmentalMicrobiology65:21842194. Poussier,S.,etal.2002.JournalofMicrobiologicalMethods51:349359. Schaad,N.W.,J.B.JonesandW.Chun.2001,3rded.LaboratoryGuideof IdentificationofPlantPathogenicBacteria.Minneapolis:APSPress. Sequeira,L.1994.Epilogue:LifewithaMutableandTreacherousTribe.In: Hayward,A.C.andG.L.Hartman.(eds.).BacterialWilt.TheDiseaseandits CausativeAgent,Pseudomonassolanacearum.Pp.235247.
Ralstonia solanacearum r 3 b 2
References-3
StatePlantRegulatoryOfficialNotification.2002.Ralstonia2002. USDAAPHISPPQISPM.Vmalik/Mneal:jhm:7348261:02/25/ 02>ralsto~1.doc. Strider,D.L.,R.K.Jones,andR.A.Haygood.1981.SouthernBacterialWiltof GeraniumCausedbyPseudomonassolanacearum.PlantDis.65:5253. Tusiime,G.,E.Adipala,F.Opio,andA.S.Bhagsari.1998.WeedsasLatent HostsofRalstoniasolanacearuminHighlandUganda:Implicationsto DevelopmentofaniNtegratedControlPackageforBacterialWilt,pages 413419.In:Prior,P.,C.Allen,andJ.Elphinstone(editors).BacterialWilt Disease:MolecularandEcologicalAspects.Berlin:SpringerVerlag Publishing;447pp. Wallace,G.B.1934.Reportofthemycologistfor1934.TanganyikaDept.Agr. Ann.Rpt.:9093. Zachos,D.G.1957.ThebrownrotofpotatoesinGreece.AnnalesdelInstitut PhytopathologiqueBenaki,NewSeries1:115117. Zehr,E.I.1969.Studiesofthedistributionandeconomicimportanceof PseudomonassolanacearumincertaincropsinthePhilippines.Philippine Agriculturist53:218223.
References-4
Ralstonia solanacearum r 3 b 2
A
Ralstonia solanacearum r 3 b 2
Appendix A
Resources
Diagnostic Tools and Equipment
CentralScienceLaboratory SandHutton York YO411LZ UnitedKingdom Telephone:+44(0)1904462000 http://www.csl.gov.uk/ LoeweBiochemicaGmbH Mhlweg2a D82054Sauerlach Germany Telephone:+49810461620 http://www.loeweinfo.com/ NeogenEuropeLimited (AcquiredAdgenLtd.) CunninghamBuilding Auchincruive Ayr,KA65HW Scotland,UK http://www.adgen.co.uk/ AgdiaIncorporated 30380CountyRoad6 Elkhart,Indiana46514USA Telephone:5742642014 Tollfree(USA):8006224342 http://www.agdia.com/ BIOREBAAG STALaboratoriesColorado 1821VistaViewDrive Longmont,CO80504 Telephone:3036516417 http://www.bioreba.ch/ http://www.stalabs.com/
Environmental Compliance
Environmental Monitoring, Categorical Exclusions
Environmental Services
FIFRA, ESA, Environmental Assessments
1
May 12, 2008 Emergency and Domestic Programs Ralstonia solanacearum r 3 b 2 A-1
Appendix A
A-2
Ralstonia solanacearum r 3 b 2
B
Ralstonia solanacearum r 3 b 2
Appendix B
Forms and Questionnaires
Contents
PPQForm391SpecimensForDetermination page B2 AddendumtoPPQForm391 page B4 QuestionnaireForNurseryAndPotatoCroppingSystemOwnersOr Managers page B5 PPQForm523EmergencyActionNotification page B7 DisposalMemorandum page B9 PPQForm526PermittoMoveLivePlantPlantsorNoxiousWeeds page B10
1
May 12, 2008 Emergency and Domestic Programs Forms.fm Ralstonia solanacearum r 3 b 2 B-1
Instructions: Type or print information requested. Press hard and print legibly when handwritten. Item 1 - assign number for each collection beginning with year, followed by collectors initials and collectors number. Example (collector, John J. Dingle): 83-JJD-001. Pest Data Section Complete Items 14, 15 and 16 or 19 or 20 and 21 as applicable. Complete Items 17 and 18 if a trap was used. 2. DATE MO DA YR State Cooperator PPQ 3. SUBMITTING AGENCY
Other
4. NAME OF SENDER
6. ADDRESS OF SENDER
INTERCEPTION SITE
ZIP
COUNTRY/ COUNTY
A.
PURPOSE
E. F. G. H.
Livestock, Domestic Animal Pest Possible Immigrant (Explain in REMARKS) Survey (Explain in REMARKS) Other (Explain in REMARKS)
B. C. D.
9. IF PROMPT OR URGENT IDENTIFICATION IS REQUESTED, PLEASE PROVIDE A BRIEF EXPLANATION UNDER REMARKS. 10. HOST INFORMATION NAME OF HOST (Scientific name when possible) NUMBER OF ACRES/PLANTS 11. QUANTITY OF HOST PLANTS AFFECTED (Insert figure and Number indicate
HOST DATA
Percent):
12. PLANT DISTRIBUTION 13. PLANT PARTS AFFECTED
Seeds
INSECTS
PUPAE ADULTS
MOLLUSKS
EGGS NYMPHS JUVS. CYSTS
PEST DATA
LARVAE
CAST SKINS
ALIVE DEAD
17. TYPE OF TRAP AND LURE 18. TRAP NUMBER
ISOLATED
20. WEED DENSITY
GENERAL
21. WEED GROWTH STAGE
FEW
22. REMARKS
SPOTTY
GENERAL
SEEDLING
VEGETATIVE
FLOWERING/FRUITING
MATURE
23. TENTATIVE DETERMINATION 24. DETERMINATION AND NOTES (Not for Field Use)
B-2
Ralstonia solanacearum r 3 b 2
OMB Information According to the Paperwork Reduction Act of 1995, no persons are required to respond to a collection of information unless it displays a valid OMB control number. The valid OMB control number for this information collection is 0579-0010. The time required to complete this information collection is estimated to average .25 hours per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing the collection of information.
Instructions
Use PPQ Form 391, Specimens for Determination, for domestic collections (warehouse inspections, local and individual collecting, special survey programs, export certification). BLOCK INSTRUCTIONS 1. Assign a number for each collection beginning the year, followed by the collectors initials and collectors number EXAMPLE
In 2001, Brian K. Long collected his first specimen for determination of the year. His first collection number is 01-BLK-001
2. Enter the collection number 2 3 4 5 6 7 8A-8L 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 Enter date Check block to indicate Agency submitting specimens for identification Enter name of sender Enter type of property specimen obtained from (farm, nursery, feedmill, etc.) Enter address Enter name and address of property owner Check all appropriate blocks Leave Blank Enter scientific name of host, if possible Enter quantity of host and plants affected Check block to indicate distribution of plant Check appropriate blocks to indicate plant parts affected Check block to indicate pest distribution Check appropriate block to indicate type of specimen Enter number specimens submitted under appropriate column Enter sampling method Enter type of trap and lure Enter trap number Enter X in block to indicate isolated or general plant symptoms Enter X in appropriate block for weed density Enter X in appropriate block for weed growth stage Provide a brief explanation if Prompt or URGENT identification is requested Enter a tentative determination if you made one Leave blank
Ralstonia solanacearum r 3 b 2
B-3
B-4
Ralstonia solanacearum r 3 b 2
Appendix B Questionnaire For Nursery And Potato Cropping System Owners Or Managers
Questions
Pleaseprovidetheinformationrequestedbelow.Youranswerswillhelp agriculturalofficialsdeterminetheextentofcontaminationinyournursery facility.Thankyouforyourcooperation. 1. Indicatethenumbersofsuspectgeraniumplantsbyvarietyand shippingdatereceivedfromtheforeignfacility,rootingstation,or wholesaler.Indicatethenameoftheshipperyoureceivedtheplants from. 2. Thecurrentlocationandnumberofallsuspectgeraniumvarietiesor shipmentsinquestion. 3. Thehistoryofmovementofsuspectgeraniumswithinthefacility. 4. Thehistoryofmovementofsuspectgeraniumsoutofthefacility.If shippedout,whichcustomerdidtheygoto?Indicatethequantityof eachvarietyandthedatesshipped. 5. Theconditionofplantssinceyoureceivedthem.Areplantsshowing wilt?Ifso,indicatethenumberofaffectedplants,andthevarieties. 6. Ifsymptomswereobservedthisseason,wherewithinthefacilitywere thewiltingplantsnoticedandwhichvariety? 7. Ifyouhaddeadorwiltedplants,whatwasthedispositionofdeador culledplantsandsoil/pottingmediaassociatedwiththoseplants?If theyweredisposedof,where?
Ralstonia solanacearum r 3 b 2
B-5
Appendix B Questionnaire For Nursery And Potato Cropping System Owners Or Managers
8. Whatwasthelocationandnumberofplantsofotherhosts(tomato, eggplant,potato,peppers)inyourfacilityinventorysincereceiving suspectshipments? 9. Whattypeofirrigationsystemdoyouuse?(i.e.,subirrigation,ebband flow,drip,handwatering,etc.)Isbackflowpreventioninplace? 10. Ifflood,subirrigation,orebbandflow,ispracticed,identifythe locationofallplantssharingthesamewatersourcewithsuspect geraniums. 11. Doyoufilterortreatyourirrigationwater?Doyouusewaterfroman outdoorholdingpondforirrigationrecirculationoroverflow? 12. Describethetypeofgreenhousebenchesusedandfloorcomposition. 13. Doyoueverstoreplantsontheground?Ifso,onwhatkindofsurface? 14. Doyoukeepyouvarietiesorshipmentsegregated? 15. Doyouuseaparticulartaggingortrackingsystemforgeraniumplants youdistribute?Ifso,pleasedescribethesystemused. 16. Doyouperformpropagationofgeraniumsfromtheplantsyoureceive fromforeignsources? 17. Whatkindofprotectiondoyournurseryworkersusewhenhandling plants?Howoftendotheywashtheirhandsorcuttingimplements? 18. Doyouuseadisinfectant?Ifso,whatisthenameofthedisinfectant? 19. Isthereastandardgreenhousesanitationprotocoldocumentavailable foryourfacility?Pleaseprovideacopyifso.
SignatureofowneroralternatecontactpersonDate
B-6
Ralstonia solanacearum r 3 b 2
Ralstonia solanacearum r 3 b 2
B-7
Instructions
UsePPQForm523EmergencyActionNotificationtoholdsuspectgeraniums andpotentiallyinfectedplantsatnurseriesorotherfacilities.Onlyoneform shouldbeusedtoholdeachlotofsuspectgeraniumsandassociated potentiallyinfectedplantmaterial.Thesameformwillbeusedtodetermine whatisheldorordereddestroyed,andwhatisreleased. Formscanbecompletedbyhand,orusetheelectronicversion.Followthe instructionsbelow.
Address http://www.aphis.usda.gov/library/forms/
Procedure
Block 1EnterthenameoflocationofthenearestPPQoffice. Block 3UnderNameofArticleenterPelargoniumspp.,notGeranium. Block 4Enterthegreenhousenumbersorotherinformationindicatingthe
locationoftheplantsheld.
Block 6Enterthesourcenurseryorforeigncountryshipper. Blocks 7 and 8Completeonlyifthatinformationisknown. Block 12ToplaceplantswithinanurseryonHold,enterfortheRalstonia solanacearumrace3biovar2asthepest.Theauthorityunderwhichactions aretakenis7CFR330andthePlantProtectionAct. Block 15ActionRequiredtextisasfollows: Allgeranium(Pelargoniumspp.)_________________________varieties receivedfrom__________________duringthedates___________________ areprohibitedfrommovementfromthenurserypropertypendingfurther notificationbyUSDAAPHISPPQ.Anyotherplantmaterialreceivedby thosesameshipmentsthatmayhavebeenexposeddirectly,inshippingor sincebeingreceived,bysharedirrigationsystems,orbyunsanitarynursery culturalpracticesarealsosubjecttothishold.Allhostplantsassociated withtheaboveexhibitingsymptomsofwiltmustbereportedimmediately toUSDAAPHISPPQandhelduntilfurthernotice.Nootherpotentialhost materialofRalstoniasolanacearumrace3biovar2mayleavegreenhouses containingsuspectplantmaterialuntilfurtherevaluationscanbemade.The abovelistedplantsandallpotentiallyinfestedmaterialafterfurther investigationsareconductedwillbedestroyedeitherbyincineration,steam sterilization,oranapprovedlandfillinaccordancewithUSDApolicies. AreashousinginfectedmaterialshallbedisinfectedaccordingtoUSDA protocols.
Releasing Plants
Block 16Afterallhostshavebeenheldatanurserywithsuspectgeraniums
B-8
Ralstonia solanacearum r 3 b 2
Disposal Memorandum
SubmittheDisposalMemorandum(Figure B7)onUSDAletterheadif approvedlandfilloperatorsrequiredocumentation.
Memorandum For Record March 28, 2003 Based on a review of available data, consultation with the U.S. Environmental Protection Agency (EPA) Office of Solid Waste Management and the APHIS Safety, Health, Environmental and Security Branch, we have determined that Ralstonia solanacearum race 3 biovar 2 does not present a risk to human health or the environment, with the exception of certain agricultural crops. We have based our conclusion on the following findings: Ralstonia solanacearum race 3 biovar 2 is not a human or animal pathogen EPA reports that Ralstonia solanacearum race 3 biovar 2 is not considered a hazardous or medical waste EPA reports that it can be disposed of in permitted solid waste It poses no threat to ground water There is a potential risk of infection for certain agricultural crops, especially potatoes but also tomatoes and peppers and other solanaceous plants It can be established in the environment through contamination of natural waterways, soil, and host plants In order to prevent the release of Ralstonia solanacearum race 3 biovar 2 to the environment and to protect at-risk crops, APHIS requires the following mitigations in its Ralstonia eradication action plan: 1. Waste (whole plants, plant material, soil and equipment) shall be: double-bagged in securely sealed, leak-proof plastic bags disposed of in a state, local or tribally permitted solid waste landfill covered to a depth of two feet with soil at landfill 2. A federal or state officials shall witness disposal at the landfill to ensure proper handling. Copies of the action plan are available at the APHIS Web site at: http://www.aphis.usda.gov/ ppq/ep/ralstonia/index.html
Ralstonia solanacearum r 3 b 2
B-9
Appendix B PPQ Form 526 Permit to Move Live Plant Plants or Noxious Weeds
PPQ Form 526 Permit to Move Live Plant Plants or Noxious Weeds
According to the Paperwork Reduction Act of 1995, no persons are required to respond to a collection of information unless it displays a valid OMB control number. The valid OMB control number for this information is 0579-0054. The time required to complete this information collection is estimated to average 0.17 hours per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing the collection of information. No permit can be issued to move live plant pests or noxious weeds until an application is received (7 CFR 330 (live plant pests) or 7 CFR 360 (noxious weeds)). U.S. DEPARTMENT OF AGRICULTURE ANIMAL AND PLANT HEALTH INSPECTION SERVICE PLANT PROTECTION AND QUARANTINE PERMITS AND RISK ASSESSMENT, UNIT 133 RIVERDALE, MARYLAND 20737
Pathogens
Arthropods
Noxious Weeds
Other (Specify) _____________________________________ This permit does not authorize the introduction, importation, interstate movement, or release into the environment of any genetically engineered organisms or products. A. SCIENTIFIC NAMES OF PESTS TO BE MOVED B.. CLASSIFICATION (Orders, Familites, Races, or Strains)
) E. SHIPPED FROM (Country or State) F. ARE PESTS G. ESTABLISHED IN U.S.? MAJOR HOST(S) OF THE PEST
4. 5. 6. 7. WHAT HOST MATERIAL OR SUBSTITUTES WILL ACCOMPANY WHICH PESTS (Indicate by line number)
8. DESTINATION
9. PORT OF ARRIVAL
12. SUPPLIER
17.
SIGNATURE OF APPLICANT (Must be person named in Item 1) Applicant must be a resident of the U.S.A. I/We agree to comply with the safeguards printed on the reverse of this form, and understand that a permit may be subject to other conditions specified in Section B and C.
18. DATE
WARNING: Any alteration, forgery, or unauthorized use of this document is subject to civil penalties of up to $250,000 (7 U.S.C. s7734(b)) or punishable by a fine of not more than $10,000, or imprisonment of not more than 5 years, or both (18 U.S.C. s1001). SECTION B - TO BE COMPLETED BY STATE OFFICIAL
19. RECOMMENDATION Concur (Approve) (Accept USDA Decision) 21. SIGNATURE 22. TITLE 23. STATE 24. DATE 20. CONDITIONS RECOMMENDED Comments (Disapprove)
PERMIT
(Permit not valid unless signed by an authorized official of the Animal and Plant Health Inspection Service) Under authority of the Plant Protection Act of 2000, permission is hereby granted to the applicant named above to move the pests described, except as deleted, subject to the conditions stated on, or attached to this application. (See standard conditions on reverse side.)
(OCT 2001)
B-10
Ralstonia solanacearum r 3 b 2
Appendix B
Ralstonia solanacearum r 3 b 2
B-11
Appendix B
B-12
Ralstonia solanacearum r 3 b 2
C
Ralstonia solanacearum r 3 b 2
Appendix C
Diagnostic Validations and Isolation Methods
Contents
Introduction page C1 Nonplate,Rapid,ImmunoassayDetection page C2 ImmunoplateAssays(ELISA) page C3 ComparisonOfNonplateImmunoassayAndPlateImmunoassay page C4 ValidationOfCentralScienceLaboratoryTaqmanAssay page C5 ValidationOfSampleAndDnaPreparationMethods page C5 PreparationOfMediaForIsolation page C6 IsolatingBacteriaFromTissueSamples page C8
Introduction
UseAppendixAtolearnmoreaboutlaboratorytestsusedtodetectthespecies Ralstoniasolanacearum. Validationstudieswereperformedonvariousdiagnostickitsformaking determinationsofRalstoniasolanacearumfromgeraniumsamples.Additional informationonisolatingthebacteriumwasalsoprovided,aswellasspecific mediarecipes. InformationinthisAppendixwasprovidedbytheUSDAAPHISPPQ CPHSTNationalPlantGermplasmandBiotechnologyLaboratory(NPGBL), Beltsville,Maryland.SeeResourcesonpage A1forinformationon contactingmanufacturersofthediagnostickits.
Important All Ralstonia solanacearum diagnostics mentioned below determine species, not biovar or race. Buffers vary by manufacturer and often produce false positive reactions when used with another manufacturers kit. Kit instructions must be followed while performing the tests. For best results, plant samples need to be symptomatic, and include the lower main stem area, or material near base of the first lateral shoot. Negative results do not necessarily indicate Ralstonia solanacearum is absent in the plant.
1 2
Recently acquired by Neogen Europe Limited. STA Laboratories, Inc., in the United States.
1
C-1
Ralstonia solanacearum r 3 b 2
Agdia ImmunoStrip
TheAgdiadeviceconsistentlydetectedRalstoniasolanacearumat107cfu/ml and,inafewtests,at106cfu/ml.Whentestinginfectedplantmaterial,thenew AgdiaImmunoStripwaseasiertovisuallyscore.Thedarkerbandcolor madelowconcentrationsofRalstoniasolanacearumeasiertoseethaneitherthe CSLorAdgendevices.TheAgdiaImmunoStriptestrequires30minutesto completewhereasboththeCSLandAdgenlateralflowkitsrequireabout3 minutes.Thiskitinstructstheusertosamplefromstems.
C-2
Ralstonia solanacearum r 3 b 2
Incubation2 Comments
1 2
Manufacturer
Adgen Ltd. Identikit 1131-09 (pre-coated) > 4802 Antibody pre-coated microplates with removable strips. Extra antibody binding step required for TAS. Several buffer dilutions required; extra fee for brokerage. Incubator required. Lacks instruction for testing bacterial samples.
150
Antibody pre-coated microplates with removable strips; no buffer dilutions; short incubations. All incubations at room temperature. Very rapid. Includes instruction for testing bacterial samples.
1620
Uncoated; antibody coating step required; 10X and 5X buffers need diluting; overnight incubation step required in step 2. Incubator required. Lacks instruction for testing bacterial samples.
1560
Uncoated; antibody coating step required; buffers must be made by user. Overnight incubation step required. Soaking is required in 3rd & 4th wash in each wash step. Incubator required. Includes instruction for testing bacterial samples.
Comments
1 2 Minutes
C-4
Ralstonia solanacearum r 3 b 2
Ralstonia solanacearum r 3 b 2
C-5
Step 2Preparebasalmedium.
How to Prepare Basal Medium 1. Per liter of dH2O add each of the following: 5g Glucose, sucrose or dextrose 10g Peptone 1g Casamino acids 12-18g Agar 2. Adjust pH of to 7.0 with KOH if necessary. 3. Autoclave 20 min. @ 121 C. 4. Cool to 55 C.
Step 3Add5mlof1%TZCstocksolutiontobasalmedium. Step 4Pour~20mlperplate. Step 5Allowtosetupovernight Step 6Incubateat28to30Cfor3648hours. Step 7Identifycolonies. TypicalRalstoniasolanacearumcoloniesonthisselectivemediumproduce irregularshaped,fluidal,mucoid,nonsymmetricalcolonies.Coloniesare typicallywhitewithpink,lightpink,orpaleredcenters.
C-6
Ralstonia solanacearum r 3 b 2
Step 4Addthefollowingstocksolutionsdissolvedin70%ethanol: 0.5ml1%Crystalviolet 2.5ml1%Bacitracin(73000units/g) 0.5ml1%Chloramphenicol(watersoluble) 0.5ml0.1%PenicillinGsodiumsalt Step 5Optional:Add2.5ml1%Cyclohexamidetoinhibitfungi. Step 6Addthefollowingfilteredsolutions(dH20): 10mlPolymyxinBSulfateSolution(8100units/mg) 5ml1%2,3,5TriphenyltetrazoliumChloride Step 7Pour~20ml/plate. Step 8Allowtodryovernight. Step 9Storeat4C. Step 10Incubateat35Cfor48hours. Step 11Identifycolonies. TypicalRalstoniasolanacearumcoloniesonthissemiselectivemediumproduce creamycolonies.RedpigmentedcoloniesarenotRalstoniasolanacearum.
Ralstonia solanacearum r 3 b 2
C-7
Method A
MethodAwassuppliedbyC.Allen(pers.comm.). Step 1Cutasmalltissuesection(~0.5cmsquare),ideallyfromstemtissue. Surfacesterilizeasectionofstem,ifdesired. Step 2Maceratein<1mlsterilewater. Step 3Allowtosit<15minutes. Step 4Streakseveralplatesdirectly,usingloopfulsofthesuspension.You maywanttousemorethanonetissuesampleandstreakplatesfromeach.
Method B
MethodBwassuppliedbyK.Rane(pers.comm.). Squeezeoutadropofcreamyoozefromasymptomaticpetioleorstemand streakitout.
Method C
MethodCwassuppliedbyS.H.Kim(pers.comm.). Step 1Locatesymptomaticleaves. Step 2Observebacterialstreamingfromleaf,oroozingfromapetioleorstem. Step 3Placeafewpetioleorstemsegments(~3mm),orfinelysliced leafblades,inatesttubecontaining10ml,0.1Mphosphatebuffer(pH7.0). Step 4Wait15minutesandstreakplates.
C-8
Ralstonia solanacearum r 3 b 2
D
Ralstonia solanacearum r 3 b 2
Appendix D
Diagnostic Laboratories
Contents
Introduction page D1 DeterminationOfRaceAndBiovar page D1 DeterminationOfGenusAndSpecies page D2
Introduction
UseAppendixBtoidentifydiagnosticlaboratories.
Important Laboratories must notify their State Plant Regulatory Official (SPRO) (http://www.aphis.usda.gov/npb/ npbmemb.html#Membership%20Dirctory) and State Plant Health Director (SPHD) (http://www.ceris.purdue.edu/napis/names/sphdXstate.html) when sending samples for race and biovar determination. States sending samples for Ralstonia solanacearum determination to other states must assure that the laboratory receiving samples has a Plant Pest Permit (PPQ Form 526) to receive those samples from out-of-state. Laboratories receiving samples from within their own states do not need permits. For more information, see: http://www.aphis.usda.gov/ppq/ permits/plantpest/index.html.
Important
1
May 12, 2008 Emergency and Domestic Programs Appendix State labs.fm Ralstonia solanacearum r 3 b 2 D-1
D-2
Ralstonia solanacearum r 3 b 2
Arizona Arkansas
California
Colorado
Connecticut
Delaware
Ralstonia solanacearum r 3 b 2
D-3
Georgia
Idaho
Indiana
Kansas
Kentucky
D-4
Ralstonia solanacearum r 3 b 2
Maryland
Massachusetts
Michigan
Ralstonia solanacearum r 3 b 2
D-5
Mississippi
Missouri
Nebraska
Nevada
New Hampshire
D-6
Ralstonia solanacearum r 3 b 2
New Mexico
New York
Ralstonia solanacearum r 3 b 2
D-7
Oregon
Pennsylvania
Tennessee Texas
D-8
Ralstonia solanacearum r 3 b 2
Virginia
Vermont
Washington
West Virginia
Wisconsin
Wyoming
Ralstonia solanacearum r 3 b 2
D-9
Colorado, Kansas, Montana, North Dakota, Oklahoma, South Dakota, northern Texas, Nebraska, Wyoming
Alabama, Louisiana, southern Texas, Arkansas, Mississippi, Virginia, Florida, North Carolina, Puerto Rico, Georgia, South Carolina, Kentucky, Tennessee
American Samoa, Arizona, California, Hawaii, Idaho, Alaska, Nevada, New Mexico, Oregon, Guam, Utah, Washington
Any state
D-10
Ralstonia solanacearum r 3 b 2
E
Ralstonia solanacearum r 3 b 2
Appendix E
Pelargonium in the N u r s e r y Tr a d e
Seed geranium
Pelargonium peltatum
Ivy geranium
Pelargonium domesticum
May 12, 2008 Emergency and Domestic Programs Appendix geraniums trade.fm
Ralstonia solanacearum r 3 b 2
E-1
Appendix E
E-2
Ralstonia solanacearum r 3 b 2
F
Ralstonia solanacearum r 3 b 2
Appendix F
Comparison of Symptoms
FIGURE F-1: Left: Early wilting caused by infection with Ralstonia solanacearum race 3 biovar 2 Right: Advanced wilting and abnormal leaf yellowing (chlorosis) caused by infection with R. solanacearum race 3 biovar 2 [Images courtesy of Wisconsin Department of Agriculture, Trade and Consumer Protection]
FIGURE F-2: Left: Abnormal yellowing caused by infection with Ralstonia solanacearum race 3 biovar 2 [Courtesy of the Plant Disease Diagnostics Clinic, University of Wisconsin-Madison, Cooperative Extension Service] Right: Close-up of wilting and necrosis caused by infection with R. solanacearum race 3 biovar 2 [Courtesy of Margery Daughtrey, Cornell University]
1
May 12, 2008 Emergency and Domestic Programs Appendix symptom.fm Ralstonia solanacearum r 3 b 2 F-1
Appendix F
FIGURE F-3: Left: Wilting and mortality of geraniums caused by infection with Ralstonia solanacearum race 3 biovar 2. Right: Mortality (top) and early wilt (right) caused by infection with R. solanacearum race 3 biovar 2
FIGURE F-4: Left: Cross-section of geranium stem with bacterial ooze caused by infection with Ralstonia solanacearum race 3 biovar 2 Right: Bacterial streaming from an infected stem immersed in water caused by infection with R. solanacearum race 3 biovar 2
F-2
Ralstonia solanacearum r 3 b 2
Appendix F
FIGURE F-5: Left: Wilting caused by bacterial blight (Xanthomonas pelargonii) is indistinguishable from Ralstonia solanacearum wilting Right: Bacterial blight (Xanthomonas pelargonii) also causes characteristic spotting of leaf tissue [Images courtesy of M. Daughtrey, Cornell University]
FIGURE F-6: Left: Early wilting symptoms on pepper caused by infection with Ralstonia solanacearum Right: Advanced wilting and necrosis of potato caused by infection with Ralstonia solanacearum [Images courtesy of CABI, 2006]
Ralstonia solanacearum r 3 b 2
F-3
Appendix F
FIGURE F-7: Left: Early tuber symptoms caused by Ralstonia solanacearum (Easily confused with ring rot caused by Clavibacter michiganense pv. sepidonicus) Right: Advanced tuber symptoms caused by R. solanacearum [Images courtesy CABI, 2006]
FIGURE F-8: Left: Bacterial streaming of infected stem immersed in water caused by infection with Ralstonia solanacearum (RS) [Image courtesy of UGA] Right: Bacterial ooze on tubers infected with R. solanacearum [Image courtesy of EMBRAPA]
F-4
Ralstonia solanacearum r 3 b 2
Appendix F
FIGURE F-9: Left and Right: Advanced ring rot of tuber caused by Clavibacter michiganense pv. sepidonicus [Left image courtesy of CABI, 2006; right image courtesy of CIP]
FIGURE F-10: Left and Right: Intermediate ring rot of tuber caused by Clavibacter michiganense pv sepidonicus [Images courtesy of Canadian Government of British Columbia and Manitoba]
Ralstonia solanacearum r 3 b 2
F-5
Appendix F
F-6
Ralstonia solanacearum r 3 b 2
G
Ralstonia solanacearum r 3 b 2
Appendix G
Disinfectants
TABLE G-1 : Disinfectants Registered Against Pseudomonas spp. (Ralstonia solanacearum = Pseudomonas solanacearum)
EPA Reg. No. 499-3 68
Active Ingredients n-Alkyl dimethyl benzyl ammonium chloride; n-Alkyl dimethyl ethylbenzyl ammonium chloride; same as Physan 20
Use Sites Work area, benches, pots, flats, flower buckets, cutting tools, greenhouse glass, walkways, evaporative coolers, decorative pools, fountains, and water displays Greenhouses, hard surfaces, lawn and turf grass, seedlings, cut flowers, decorative fountains, pools, birdbaths, and plants Greenhouse structures, benches, pots, watering systems, evaporative coolers, storage rooms, ventilation equipment, floors and other equipment Greenhouse hard surfaces, work areas, benches, flower pots, buckets, flats, cutting tools, walkways, garden bird baths, and evaporative coolers Greenhouse hard surfaces, work areas, benches, cutting tools, walkways, garden bird baths, and evaporative coolers Non-porous, inanimate surfaces such as floors, walls, metal surfaces, stainless steel surfaces, plastic surfaces, knobs, handles, and railings
Physan 20
5536 4-5
Same as Green-Shield
Pseudomonas spp.
Zero TolZT
7029 9-1
Hydrogen dioxide
Pseudomonas spp.
5804 4-3
n-Alkyl dimethyl benzyl ammonium chloride; n-Alkyl dimethyl ethylbenzyl ammonium chloride; same as Triathlon Same as Consan Triple Action 20
Triathlon
AE-90
4737 1-89
n-Alkyl dimethyl benzyl ammonium chloride; n-Alkyl dimethyl ethylbenzyl ammonium chloride
Pseudomonas aeruginosa
1
G-1
Ralstonia solanacearum r 3 b 2
Appendix G
TABLE G-1 : Disinfectants Registered Against Pseudomonas spp. (Ralstonia solanacearum = Pseudomonas solanacearum)
Lonza S-18 683677 Octyl decyl dimethyl ammonium chloride; Dioctyl dimethyl ammonium chloride; didecyl dimethyl ammonium chloride; n-Alkyl dimethyl benzyl ammonium chloride Octyl decyl dimethyl ammonium chloride; Dioctyl dimethyl ammonium chloride; didecyl dimethyl ammonium chloride; n-Alkyl dimethyl benzyl ammonium chloride Octyl decyl dimethyl ammonium chloride; Dioctyl dimethyl ammonium chloride; didecyl dimethyl ammonium chloride; n-Alkyl dimethyl benzyl ammonium chloride Octyl decyl dimethyl ammonium chloride; Dioctyl dimethyl ammonium chloride; didecyl dimethyl ammonium chloride; n-Alkyl dimethyl benzyl ammonium chloride; same as MAQUAT 615-LR Same as MAQUAT 615-HD Farm, poultry, swine, and mushroom premise sanitation; veterinary practice, animal care, animal laboratory disinfection Pseudomonas aeruginosa and Pseudomonas cepacia
MAQUAT 128-MT
1032 4-112
Outer clothing, field harvesting equipment, walls/floors of coolers, flower buckets, and greenhouse packing areas
Pseudomonas aeruginosa
MAQUAT 64 MN
1032 4-113
Florist shops, wholesale florists, shippers, greenhouse packing areas, flower buckets, floors/walls of coolers, design and packaging benches, and counter tops Greenhouses, hard non porous surfaces (flower buckets, floors, walls, coolers, design, packing benches and counter tops)
Pseudomonas aeruginosa
MAQUAT 615-HD
1032 4-72
MAQUAT 615-LR
1032 4-109
G-2
Ralstonia solanacearum r 3 b 2