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Daniel FloresFebruary 3, 2010
Methods Development for Mass Production of Tamarixia radiata
Dan Flores, PhDUSDA APHIS PPQ CPHST
Mission Laboratory
Daniel FloresFebruary 3, 2010
PRODUCTION
• Arthropod QuarantinePakistan Strain
• Lab ProductionTaiwan / Vietnam Strain
• Field ProductionTaiwan / Vietnam Strain
Daniel FloresFebruary 3, 2010
MOVEMENT INTO U.S.
• July 10, 2009 - Leeda Wood (Arthropod Quarantine Officer) contacts Abdul Rehman(Insect Biocontrol Specialist - CABI South Asia)
• July 29, 2009 - Permit P526P-09-02428 approved
• August 8, 2009 - First shipment dispatched
• August 11, 2009 – Shipment received and cleared at Los Indios Plant Inspection Station
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TAMARIXIA COLLECTIONS
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SHIPMENTS RECEIVEDPPQ
Receiver’s ID No.
DateSent
Date Received
# of Pupae (Mummies)
PPQMTX081109 August 8, 2009 August 11, 2009 700
PPQMTX081409 August 12, 2009 August 14, 2009 700
PPQMTX082009 August 18, 2009 August 20, 2009 600
PPQMTX090309 August 31, 2009 September 3, 2009 375
TOTAL 2375
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ARTHROPOD QUARANTINE
• 12 environmental growth chambers• 5 greenhouses• 4 lab areas• 1 office• 1 headhouse• 1 pass-thru chamber• 1 receiving room• 1 autoclave pass-thru
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QUARANTINE ENTRY
Pass-thru Chamber
Secondary Containment Cage in Receiving Room
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COLLECTION VIALS
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PUPAE
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VARIOUS STAGES
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PENNY FOR YOUR THOUGHTS
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REARING CAGE IN EGC
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ENCYRTIDAEDiaphorencyrtus aligarhensis
• Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)
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SIGNIPHORIDAEChartocerus sp.
• Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)
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EULOPHIDAEAprostocetus sp.
• Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)
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• Dr. Gregory Evans (USDA / Systematic Entomology Laboratory, Beltsville, MD)• Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)• Dr. Norman B. Barr (USDA APHIS PPQ CPHST Mission Laboratory, Edinburg, TX)
EULOPHIDAE: Tamarixia radiata
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ISO-FEMALE LINES6” Pots
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QUALITY CONTROL
• 12-16 cages / EGC x 2 chambers
• Specimens submitted weekly
• Collect and submit 20 adult specimens from 4 random cages (5 / cage)
• Results - 6 haplotypes
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LAB PRODUCTION AREAS
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MAIN GREENHOUSE
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ORANGE JASMINE
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ADULT PSYLLID COLLECTIONS
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SHADEHOUSE PRODUCTION
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ADULT PSYLLIDS
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ENVIRONMENTAL GROWTH CHAMBERS
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28oC, 50% RH, 14:10 L:D
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PSYLLID COLONY
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200-400 PSYLLIDS / CAGE
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INFESTED PLANT
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PARASITE CHAMBER
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TAIWAN / VIETNAM STRAIN
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PARASITOID COLLECTION
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ISO-FEMALE CYLINDERS
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EMERGENCE CAGES
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ONE PLANT PER CAGE
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PARASITOID PRODUCTION
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STICKY TRAP ON PETRI-DISH
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TOTAL PARASITOID EMERGENCE
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HOST FEEDING STUDY
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CONE-TAINERS
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HOST MORTALITY
35.26%
41.19%
23.55%
n=28 Tamarixia radiata females
ACP
T.radiata
Host-fed
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LARGE ORANGE JASMINE PLANT
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PLANT PRUNED TO INDUCE UNIFORM FLUSH
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PSYLLIDS EGGING PLANT
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~2000 EGGS / PLANT
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COLLECTION METHODS
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PROJECTIONS400 parasitoids / plant x 25 plants / week10,000 parasitoids / weekX 4 weeks40,000 parasitoids / month
1000 parasitoids / plantX 25 plants / week25,000 parasitoids / weekX 4 weeks100,000 parasitoids / month
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FIELD PRODUCTION AT USDA CITRUS GROVE
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TREE BEING CLIPPED TO INDUCE FLUSH
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FRAME INSTALLED
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ABUNDANCE OF FLUSH
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FIELD CAGE FOR MASS PRODUCTION
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FIELD INSECTARY CAGES
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PSYLLID PRODUCTION
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PARASITOID RELEASES
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COLLECTION TRAP
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CURRENT PRODUCTION
• during the months of October-November
• low-volume release of 20 parasitoids
• 2-3 generations
• parasitism rates of 19.8% were observed
• estimated 2000 parasitoids
• net return of 100X
Daniel FloresFebruary 3, 2010
PROJECTIONS
• increase our yield
• timing - in the spring and summer months
• release numbers - 10 to 50-fold
• inducing 2 to 5-times more flush
• project 40,000 - 200,000 parasitoids per tree
Daniel FloresFebruary 3, 2010
• Quarantine screening, rearing, and mass production of new strains of Tamarixia and any other biological control agents.
• Develop methods and protocols for the easy release of large volumes of biological control organisms.
• Qualify and quantify the impact of large-volume releases of biological control agents in organic groves, commercial groves, and dooryards.
• Transfer technology and/or biological control organisms to federal, state, and other cooperators in affected states for release.
FUTURE GOALS
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END
Daniel FloresFebruary 3, 2010
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