Roadkill Survey

The main objectives of the study are to -
1. Establish a surveillance program for documenting pathogen presence in roadkill.
2. Identify roadkill hotspots for targeted conservation efforts.
Standard Operating Procedure (SOP)
• Equipment Checklist:
o Cryo vials 50
o Swabs 50
o Rnalater 25 (125)
o Cryo boxes 2
o Scalpel
o Blades 100 pk
o Tissue forceps 2
o Artery forcep 2
o Scissors 2
o Gloves 1 box x 3
o Marker 2 fine tip
o Ice box
o Ice packs 2
o Mask
o Sanitizers
o Discards box
o Absolute alcohol
o Cotton
• Drive at recommended speed (40–50 km/h) to maximize visibility and minimize missed carcasses
• On spotting a carcass, indicate, slow, and safely pull over to the roadside
• Park fully off the road with hazard lights on
• Fill Epicollect5 form
• Fill Roadkill app online labelling details
• Inform the Concerned Wildlife Health Officer
• Assess traffic before exiting vehicle
• Quickly visually scan the surroundings for additional carcasses, live animals or hazards
• Surveyor must always remain alert to traffic
4. Carcass Assessment and Sample Collection
• Record details as mentioned in the field data sheet and labelling app
• Photograph carcass
• Sampling Order:
• A. Swabs: Collect swabs as indicated in the field data sheet.
Use new sterile tools and swabs for each sample; avoid cross-contamination
• B. Brain Tissue:
Expose the foramen magnum through dorsal incision at the base of skull
Take tissues through the brain stem using a straw/ scoop.
5. Sample Preservation
• Brain Tissue:
For RT-PCR:
Place portion directly in sterile, RNase-free tube—freeze at −20°C or below as soon as possible
If freezing cannot occur immediately, store on ice packs for up to 24 hrs
• Swabs:
Place each swab in tube with RNALater
Store on ice packs in icebox for up to 24 hrs