Methods Development for Mass Production of Tamarixia radiata. Dan Flores, PhD USDA APHIS PPQ CPHST Mission Laboratory

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1 Methods Development for Mass Production of Tamarixia radiata Dan Flores, PhD USDA APHIS PPQ CPHST Mission Laboratory Daniel Flores

2 PRODUCTION Arthropod Quarantine Pakistan Strain Lab Production Taiwan / Vietnam Strain Field Production Taiwan / Vietnam Strain Daniel Flores

3 MOVEMENT INTO U.S. July 10, Leeda Wood (Arthropod Quarantine Officer) contacts Abdul Rehman (Insect Biocontrol Specialist - CABI South Asia) July 29, Permit P526P approved August 8, First shipment dispatched August 11, 2009 Shipment received and cleared at Los Indios Plant Inspection Station Daniel Flores

4 TAMARIXIA COLLECTIONS

5 SHIPMENTS RECEIVED PPQ Receiver s ID No. Date Sent Date Received # of Pupae (Mummies) PPQMTX August 8, 2009 August 11, PPQMTX August 12, 2009 August 14, PPQMTX August 18, 2009 August 20, PPQMTX August 31, 2009 September 3, TOTAL 2375 Daniel Flores

6 ARTHROPOD QUARANTINE Daniel Flores 12 environmental growth chambers 5 greenhouses 4 lab areas 1 office 1 headhouse 1 pass-thru chamber 1 receiving room 1 autoclave pass-thru

7 QUARANTINE ENTRY Pass-thru Chamber Secondary Containment Cage in Receiving Room Daniel Flores

8 COLLECTION VIALS

9 PUPAE

10 VARIOUS STAGES

11 PENNY FOR YOUR THOUGHTS

12 REARING CAGE IN EGC

13 ENCYRTIDAE Diaphorencyrtus aligarhensis Daniel Flores Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)

14 SIGNIPHORIDAE Chartocerus sp. Daniel Flores Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)

15 EULOPHIDAE Aprostocetus sp. Daniel Flores Dr. Michael Gates (USDA / Systematic Entomology Laboratory, Beltsville, MD)

16 EULOPHIDAE: Tamarixia radiata Daniel Flores 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)

17 ISO-FEMALE LINES 6 Pots

18 QUALITY CONTROL cages / EGC x 2 chambers Specimens submitted weekly Collect and submit 20 adult specimens from 4 random cages (5 / cage) Results - 6 haplotypes Daniel Flores

19 LAB PRODUCTION AREAS

20 MAIN GREENHOUSE

21 ORANGE JASMINE

22 ADULT PSYLLID COLLECTIONS

23 SHADEHOUSE PRODUCTION

24 ADULT PSYLLIDS

25 ENVIRONMENTAL GROWTH CHAMBERS Daniel Flores

26 28 o C, 50% RH, 14:10 L:D

27 PSYLLID COLONY

28 PSYLLIDS / CAGE

29 INFESTED PLANT

30 PARASITE CHAMBER

31 TAIWAN / VIETNAM STRAIN

32 PARASITOID COLLECTION

33 ISO-FEMALE CYLINDERS

34 EMERGENCE CAGES

35 ONE PLANT PER CAGE

36 PARASITOID PRODUCTION

37 STICKY TRAP ON PETRI-DISH

38 TOTAL PARASITOID EMERGENCE Daniel Flores

39 HOST FEEDING STUDY

40 CONE-TAINERS

41 HOST MORTALITY 23.55% 41.19% 35.26% ACP T.radiata Host-fed n=28 Tamarixia radiata females Daniel Flores

42 LARGE ORANGE JASMINE PLANT Daniel Flores

43 PLANT PRUNED TO INDUCE UNIFORM FLUSH Daniel Flores

44 PSYLLIDS EGGING PLANT

45 ~2000 EGGS / PLANT

46 COLLECTION METHODS

47 PROJECTIONS 400 parasitoids / plant x 25 plants / week 10,000 parasitoids / week X 4 weeks 40,000 parasitoids / month 1000 parasitoids / plant X 25 plants / week 25,000 parasitoids / week X 4 weeks 100,000 parasitoids / month Daniel Flores

48 FIELD PRODUCTION AT USDA CITRUS GROVE Daniel Flores

49 TREE BEING CLIPPED TO INDUCE FLUSH Daniel Flores

50 FRAME INSTALLED

51 ABUNDANCE OF FLUSH

52 FIELD CAGE FOR MASS PRODUCTION Daniel Flores

53 FIELD INSECTARY CAGES

54 PSYLLID PRODUCTION

55 PARASITOID RELEASES

56 COLLECTION TRAP

57 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 Flores

58 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 parasitoids per tree Daniel Flores

59 FUTURE GOALS 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.

60 END Daniel Flores

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