1. National Plant Diagnostic Network Submitting Diagnostic Samples: Quality and Security Adapted from “Module 3 : Submitting Diagnostic Samples” by Creswell, Thomas, Cullen, Buss, Hodges, Harmon, Wright, and Ailshie. December 2006. Update, August 2008. NPDN Publication No. 0010 Edited and presented by Gail Ruhl, Purdue University First Detector Webinar Training October 14,2008

2. Submitting Diagnostic Samples Sample Security  Communication: Early contact with diagnostic labs and regulatory officials  Delivery details: Where, How, When  Confidentiality  Accuracy of source data  Chain of custody

3. Submitting Diagnostic Samples Sample Quality Diagnosis or ID is only as good as the information provided. Fill out the clinic form fully Diagnosis or ID is only as good as the sample provided. Representative sample Digital images Fresh and in good condition Rapid delivery may be critical

4. Submitting Diagnostic Samples Samples Must Contain the Correct Plant ‘Part’ Lesions / Damage on stem? Foliage diseases Root problems

5. Submitting Diagnostic Samples When the plant is too big to submit:  Include of the site and symptoms observed  Include digital images of the site and symptoms observed  Include affected branches with healthy and diseased tissue  Include feeder roots and soil

6. Submitting Diagnostic Samples Field patterns may be clues to: Chemical injury? Nematodes? Root diseases? Corn: Herbicide Injury Tobacco: Phytophthora root/stem rot Photo: Clemson University - USDA Cooperative Extension Slide Series, www.forestryimages.org Photo: Tom Jordan, Purdue University

7. Submitting Diagnostic Samples Incidence VS. Severity Incidence: Percent of the crop affected Severity: a measure of impact on a plant or the crop Photo: Andrej Kunca, National Forest Centre - Slovakia, www.forestryimages.org Photo: Clemson University - USDA Cooperative Extension Slide Series, www.forestryimages.org

8. Submitting Diagnostic Samples Dead Plants Tell no Tales  Avoid dead plants  Choose plants which show a range of symptoms: moderate to severe Photo: Ron Jones, North Carolina State University, www.forestryimages.org

9. Submitting Diagnostic Samples Sample Quality When collecting samples: Keep samples fresh in a cooler with ice packs Have plastic bags and ties Bring labels and permanent markers

10. Submitting Diagnostic Samples Sample Quality: Packaging & Shipping  Keep soil on roots  No extra water  Wrap in dry paper then double bag in plastic  Disinfest exterior of bags  Strong crush-proof box, tape all seams

11. Submitting Diagnostic Samples Packaging and Shipping Keep soil on roots –not on foliage

12. Submitting Diagnostic Samples Good Packaging  Plastic bag to keep soil on roots  Dry paper towels to protect leaves from contact with plastic bag Photos: Tom Creswell, North Carolina State University, www.forestryimages.org

13. Submitting Diagnostic Samples Packaging and Shipping Blunde rs Soil on foliage during shipping creates ‘diseases’ that were not there when the sample was collected. Photos: Tom Creswell, North Carolina State University, www.forestryimages.org

14. Submitting Diagnostic Samples Good Packaging Paper or packing tape used to cover the surface of the pot to keep soil contained during shipment to a diagnostic lab Photo: Gail Ruhl,Purdue Univeristy

15. Packaging & Shipping Good Intentions Actual Results Photos: Tom Creswell, North Carolina State University Submitting Diagnostic Samples

16. Proper Packaging Crush-Proof Box Photos: Gail Ruhl, Purdue University Submitting Diagnostic Samples

17. Secure Packaging Suspect select agents, invasives and exotics should be double bagged.

18. Submitting Diagnostic Samples Who? - Who sent the sample? Client contact information? What? What is the host? Your main concern? Symptoms observed? When? When did it show up? Where? Where in the field, Distibution, % affected? How? How has the site been treated? Pesticides, Fertilizer, Rainfall… What Every Clinic Wants to Know….

19. Submitting Diagnostic Samples Sample Delivery Client Extension Educator Postal Service External Expert(s) Diagnostician Regional Center

20. Submitting Diagnostic Samples Digital Images  Digital images can assist with secure identification  Digital images may allow for rapid detection of possible suspect exotic agents

21. Submitting Diagnostic Samples What Every Entomologist Wants to Know…..  Who collected the sample? Include their contact information.  Location on Host Plant? Roots, Stems, Flowers, Buds, Leaves, etc.  Where was the insect found?  Field Crops, Greenhouse, Residence, Landscape  Location: Address, Nearest intersection, GPS data  Degree of infestation, extent of damage

22. Submitting Diagnostic Samples Insect Samples Poor Packaging = Body Parts! Properly Packaged Mailing Tubes Protect Samples! Tubes Protect Samples! Photos: Lyle Buss, University of Florida Photo: Tom Creswell, North Carolina State University

23. Submitting Diagnostic Samples Insect Samples Most insects can be preserved in a vial with 70% isopropyl (rubbing) or ethyl alcohol. Photo: Scott Bauer, USDA Agricultural Research Service, www.forestryimages.org Photo: Tom Creswell, North Carolina State University Shipping restrictions may occur. See notes section or contact your local lab.

24. Submitting Diagnostic Samples Insect Samples  Caterpillars should be placed in boiling water for 1 minute prior to preservation.  Don’t Microwave  Include some caterpillars live on host plant foliage. Bottom Photo: Clemson University - USDA Cooperative Extension Slide Series, www.forestryimages.org Top Photo:Gerald J. Lenhard, Bugwood.org

25. Submitting Diagnostic Samples Insect Samples  Scales, mealybugs and other tiny arthropods may be submitted on the host.  Wrap plant material in dry paper towel before placing in bag.  Double bag suspected exotics. Photos: Florida Department of Agriculture, Division of Plant Industry

26. Submitting Diagnostic Samples Insect Samples  Collect multiple samples of all available life stages.  May need extra samples available if a new record.  May need specific life stage or both sexes for ID.  Which whitefly life stage is used for ID? <<< Pupa Photos: Jim Baker, North Carolina State University

27. Submitting Diagnostic Samples Insect Samples  Digital photos of damage and insect assist with identification.  Describe the level of infestation on the plant. Top Photo: Tom Creswell, North Carolina State University, www.forestryimages.org Bottom Photo: Gail Ruhl,Purdue University

28. Submitting Diagnostic Samples Piercing/Sucking Boring Leaf miningSkeletonizing Photo: James Solomon, USDA Forest Service, www.forestryimages.org Photo: Clemson University - USDA Cooperative Extension Slide Series, www.forestryimages.org Photo: E. Bradford Walker, Vermont Department of Forests, Parks and Recreation, www.forestryimages.org Photo: William M. Ciesla, Forest Health Management International, www.forestryimages.org

29. Submitting Diagnostic Samples Insect Samples: DOs  Collect multiple samples  Collect portion of the infested plant  Send Pictures  Preserve as needed  Ship quickly  Exotics? notify specialist, ship next day air  Include complete and accurate collection data  Double bag specimens containing suspected exotic species.

30. Submitting Diagnostic Samples Insect Samples: DON’Ts  Crush samples in tissue or plastic wrap  Tape them to paper  Overcrowd them (dead or alive)  Send them without complete and accurate information  Collect broken body parts

31. Submitting Diagnostic Samples Weed and Plant ID  Also for ID of insect or disease host (if unknown)  Collect intact specimens.  Preserve and package sample properly.  Exotics? Seal box inside and out, double bag. Photo: Jeffrey Mullahey, University of Florida, www.forestryimages.org

32. Submitting Diagnostic Samples Weed and Plant ID  Include: Flowers, Fruits, Stems, Leaves, Roots  May preserve samples by pressing and drying in newspaper Photos: Florida Department of Agriculture, Division of Plant Industry

33. Submitting Diagnostic Samples stems roots if possible whole leaves attached to the stem flowers, fruits, or seeds. Weeds: We want it all All Photos: John D. Byrd, Mississippi State University, www.forestryimages.org

34. Submitting Diagnostic Samples Weeds: What Every Botanist Wants to Know……  Where did it come from?  Pasture? Greenhouse? Home landscape? Field?  Location: Address, Nearest intersection, GPS data  Digital photos can be useful if they are close-ups and very clear.  Be specific on collection information! The more information you can give, the better, and faster, the diagnosis will be.  Where was the sample found?

35. Submitting Diagnostic Samples Secure and Quality Sample Submissions HELP US TO HELP YOU With the Identification of High Consequence Pathogens and Pests of Concern

36. Submitting Diagnostic Samples Tom Creswell, North Carolina State University Carla Thomas, University of California Richard Cullen, University of Florida Lyle Buss, University of Florida Amanda Hodges, SPDN, University of Florida Carrie Harmon, SPDN, University of Florida Kathy Wright, Kansas State University Tray Ailshie, Kansas State University Author Information

37. Submitting Diagnostic Samples This publication can be used for non-profit, educational use only purposes. Photographers retain copyright to photographs or other images contained in this publication as cited. This material was developed as a topic-based training module for NPDN First Detector Training. Authors and the website should be properly cited. Images or photographs should also be properly cited and credited to the original source. Publication Number: 0010 Publication Date: December 2006 Publication Details