Protocols

GLIDE Litter Extraction Procedures

BARCODE PROTOCOLS

Barcodes have been placed onto the field data sheets prior to shipment. Attach barcodes to vials holding the extracted material corresponding to the litterbags, treatment or control.

TREATMENT LITTER BAGS

  1. There are 8 barcodes for each sampling interval, 2 for each plot number.
  2. Place the 2 corresponding barcodes onto the corresponding vial that will hold the extracted fauna.

e.g. for Tasmania, sampling interval 1, plot no. 4. treatment = barcode TAS14T.

Two barcodes are required on the vial so that if one barcode comes off the vial the other one will still identify the material.

LITTERBAG COLLECTION AND PROCESSING

  1. Litterbags should be photographed prior to collection
  2. The surface of bags should be carefully cleaned of adhering soil, living plant parts (roots or moss), rock fragments, etc., immediately before collection.
  3. Retrieved bags will be placed gingerly in a plastic ziplock bag, sealed, and carefully (without much disturbance) returned to the laboratory. It is crucial to ensure that decomposing materials do not fragment and fall out of the bags during retrieval or before processing. Any fragmentation or loss of material must be documented on the data sheet for each sample.
  4. All bags should immediately be weighed to a thousandth of a gram and then placed gently on a Tullgren funnel (see extraction procedure below).
  5. Following each faunal extraction, treated litter should be oven dried at 55°C until the mass is stable, weighed to the nearest thousandth of a gram, then shipped to the NREL for appropriate chemical analyses as outlined in Harmon et al (1999) and archived on-site due to quarantine restrictions.

LITTER EXTRACTION PROTOCOLS

  1. Install funnel by joining the black laminated circle at the dotted line.
  2. Ensure that the dull side is the innermost side (this is the most waterproof side in case litter is wet).
  3. Fix with adhesive tape along the join.
  4. Join white cardboard with adhesive tape at dotted line. This makes the collar for the funnel to sit in.
  5. Turn funnel upside down, sit collar onto funnel and secure funnel to collar with adhesive tape. When turned right side up the funnel should be stable and level in the collar.
  6. Fill vial with 95% alcohol.
  7. Attach barcodes to vial.
  8. Place vial on a stable surface.
  9. Place the funnel and collar over the vial so that the hole in the funnel is directly over the vial.
  10. Place litter bag horizontally into funnel top.
  11. Bag does not need to be cut open (what crawled in will crawl back out).
  12. Place light (40watt incandescent globe) directly above funnel 19cm above litter bag.
  13. Leave light on and in position for 5 days.
  14. After 5 days remove the light and funnel, place lid on vial.
  15. Send all extracted material to BioTrack.
  16. Label parcel for customs as DEAD INVERTEBRATES IN ALCOHOL.

GLIDE Invertebrate Extraction Set-up

GLIDE datasheet and vial with barcodes (18,933 bytes) Close-up of barcodes (11,803 bytes) Sample vial with barcodes (5,597 bytes) Joining the funnel (11,735 bytes)
Collar with sample vial inside (6,168 bytes) Inverted funnel and collar (11,864 bytes) The completed set-up (20,461 bytes)