ExoticBOL: Identifying Insect Pests at US Ports of Entry

Author: Robyn Tse, Massapequa High School
Mentor: Mark Stoeckle, Program for the Human Environment, Rockefeller University
Winner, Honorable Mention, Urban Barcode Project, June 2012

INTRO

Prompt recognition of agricultural pests is imperative to prevent economic and ecological damage

  • U.S. Customs and Border Protection intercepts 40,000 to 50,000 agricultural pests yearly
  • Morphological identification often slow and inexact
    • 59% of urgent and 95% of non-urgent identifications require 2 or more days
    • Many specimens identified only to order, family, or genus

HYPOTHESIS

DNA barcoding can supplement or substitute for entomological expertise in identifying insect pests

METHODS

  • DNA extraction: Wizard Genomic DNA Purification Kit
  • Amplification: Folmer primers with M13 tails
    Forward 5’-TGT AAA ACG ACG GCC AGT GGT CAA CAA ATC ATA AAG ATA TTG G-3’
    Reverse: 5’-CAG GAA ACA GCT ATG AC TAA ACT TCA GGG TGA CCA AAA AAT CA-3’
    35 cycles of 95°C for 30 s, 50°C for 30 s, and 72°C for 45 s
  • DNA barcode ID: ≥ 95% identity to reference sequence in GenBank or Barcode of Life Datasystems (BOLD)

RESULTS

  • About 1/3 of specimens yielded DNA barcodes [10/32 (31%) yielded barcodes]
  • Yield higher from immature vs adult specimens [7/9 immatures vs 3/23 adults (p=0.001)]
  • Most gave barcode IDs [7/10 (70%) were ≥ 95% identical to a GenBank or BOLD reference sequence]
  • Barcode IDs were the same or more detailed than morphologic IDs

CONCLUSION

DNA barcoding is a potential aid to rapid, accurate identification of insect pests at U.S. ports of entry

NEXT STEPS

  • Improve barcode recovery (test alternate extraction methods, primers, storage conditions)
  • Improve reference libraries (many pest species currently not represented)

HONORS

ExoticBOL was awarded Honorable Mention at the Urban Barcode Project (UBP) award ceremony held at American Museum of Natural History, June 2012 (ExoticBOL abstract, photos, and video presentation are posted on UBP site)

Download Powerpoint

ACKNOWLEDGMENTS

U.S. Customs and Border Protection, Newark International Airport
Frances Krim Memorial Inspection Station, APHIS, USDA
DNA Learning Center, Cold Spring Harbor Laboratories
Harlem DNA Laboratory
GenSpace
American Museum of Natural History
The Alfred P. Sloan Foundation

URL: http://phe.rockefeller.edu/barcode/exoticbol.html
Last updated: Wednesday, 22-Aug-2012 18:37:41 EDT