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Daniel FloresFebruary 3, 2010
Methods Development for Mass Production of Tamarixia radiata
Dan Flores, PhDUSDA APHIS PPQ CPHST
Mission Laboratory
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Daniel FloresFebruary 3, 2010
PRODUCTION
• Arthropod QuarantinePakistan Strain
• Lab ProductionTaiwan / Vietnam Strain
• Field ProductionTaiwan / Vietnam Strain
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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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Daniel FloresFebruary 3, 2010
TAMARIXIA COLLECTIONS
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Daniel FloresFebruary 3, 2010
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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Daniel FloresFebruary 3, 2010
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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Daniel FloresFebruary 3, 2010
QUARANTINE ENTRY
Pass-thru Chamber
Secondary Containment Cage in Receiving Room
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Daniel FloresFebruary 3, 2010
COLLECTION VIALS
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Daniel FloresFebruary 3, 2010
PUPAE
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Daniel FloresFebruary 3, 2010
VARIOUS STAGES
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Daniel FloresFebruary 3, 2010
PENNY FOR YOUR THOUGHTS
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Daniel FloresFebruary 3, 2010
REARING CAGE IN EGC
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Daniel FloresFebruary 3, 2010
ENCYRTIDAEDiaphorencyrtus aligarhensis
• Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)
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Daniel FloresFebruary 3, 2010
SIGNIPHORIDAEChartocerus sp.
• Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)
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Daniel FloresFebruary 3, 2010
EULOPHIDAEAprostocetus sp.
• Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)
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Daniel FloresFebruary 3, 2010
• 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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Daniel FloresFebruary 3, 2010
ISO-FEMALE LINES6” Pots
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Daniel FloresFebruary 3, 2010
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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Daniel FloresFebruary 3, 2010
LAB PRODUCTION AREAS
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Daniel FloresFebruary 3, 2010
MAIN GREENHOUSE
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Daniel FloresFebruary 3, 2010
ORANGE JASMINE
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Daniel FloresFebruary 3, 2010
ADULT PSYLLID COLLECTIONS
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Daniel FloresFebruary 3, 2010
SHADEHOUSE PRODUCTION
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Daniel FloresFebruary 3, 2010
ADULT PSYLLIDS
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Daniel FloresFebruary 3, 2010
ENVIRONMENTAL GROWTH CHAMBERS
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Daniel FloresFebruary 3, 2010
28oC, 50% RH, 14:10 L:D
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Daniel FloresFebruary 3, 2010
PSYLLID COLONY
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Daniel FloresFebruary 3, 2010
200-400 PSYLLIDS / CAGE
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Daniel FloresFebruary 3, 2010
INFESTED PLANT
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Daniel FloresFebruary 3, 2010
PARASITE CHAMBER
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Daniel FloresFebruary 3, 2010
TAIWAN / VIETNAM STRAIN
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Daniel FloresFebruary 3, 2010
PARASITOID COLLECTION
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Daniel FloresFebruary 3, 2010
ISO-FEMALE CYLINDERS
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Daniel FloresFebruary 3, 2010
EMERGENCE CAGES
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Daniel FloresFebruary 3, 2010
ONE PLANT PER CAGE
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Daniel FloresFebruary 3, 2010
PARASITOID PRODUCTION
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Daniel FloresFebruary 3, 2010
STICKY TRAP ON PETRI-DISH
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Daniel FloresFebruary 3, 2010
TOTAL PARASITOID EMERGENCE
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Daniel FloresFebruary 3, 2010
HOST FEEDING STUDY
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Daniel FloresFebruary 3, 2010
CONE-TAINERS
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Daniel FloresFebruary 3, 2010
HOST MORTALITY
35.26%
41.19%
23.55%
n=28 Tamarixia radiata females
ACP
T.radiata
Host-fed
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Daniel FloresFebruary 3, 2010
LARGE ORANGE JASMINE PLANT
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Daniel FloresFebruary 3, 2010
PLANT PRUNED TO INDUCE UNIFORM FLUSH
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Daniel FloresFebruary 3, 2010
PSYLLIDS EGGING PLANT
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Daniel FloresFebruary 3, 2010
~2000 EGGS / PLANT
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Daniel FloresFebruary 3, 2010
COLLECTION METHODS
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Daniel FloresFebruary 3, 2010
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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Daniel FloresFebruary 3, 2010
FIELD PRODUCTION AT USDA CITRUS GROVE
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Daniel FloresFebruary 3, 2010
TREE BEING CLIPPED TO INDUCE FLUSH
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Daniel FloresFebruary 3, 2010
FRAME INSTALLED
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Daniel FloresFebruary 3, 2010
ABUNDANCE OF FLUSH
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Daniel FloresFebruary 3, 2010
FIELD CAGE FOR MASS PRODUCTION
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Daniel FloresFebruary 3, 2010
FIELD INSECTARY CAGES
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Daniel FloresFebruary 3, 2010
PSYLLID PRODUCTION
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Daniel FloresFebruary 3, 2010
PARASITOID RELEASES
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Daniel FloresFebruary 3, 2010
COLLECTION TRAP
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Daniel FloresFebruary 3, 2010
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
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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
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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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Daniel FloresFebruary 3, 2010
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Daniel FloresFebruary 3, 2010