Protocol - Swabbing
@Zerina Rahic
Swabbing
- Recommended to sample at least 20 to 30 individuals from a meta population in order to detect Bd if present
- Recommended to analyse each sample separately - potential increase in cost
- Pooling should be done in the lab not in the field
- It is advisable to sample species during breeding period
Material
-70% EtOH - cotton tipped applicators (“swabs”) - vials (~2ml w/ self-sealing screw caps microcentrifuge tubes are recommended) - powder-free latex or nitrile gloves - alcohol sanitizer for hands - water- and alcohol-proof pens - Waterproof notebook - sharp-pointed dissecting probe - waste bag or container - collection bags - closeable bags - vial storage boxes - Bleach solution: 1 part commercial bleach (5% sodium hypochlorite) to 9 parts water
Procedure:
Preparation: 1. Equipment check, a thorough checklist is advised 2. Organize equipment in field vest(s) for easy access 3. Prepare data pages to include data to collect 4. Begin survey 5. Person 1 puts on clean gloves to capture amphibian 6. Person 2 put on clean gloves in case of accidental contamination of hands, and readies all equipment needed (see list) 7. Walk transect to visually encounter amphibians Capture: 1. Only Person 1 should attempt to capture amphibian 2. Attempt to capture amphibian as efficiently as possible 3. Minimize handling time for the amphibian, and minimize any contact you may have with any substrate. A clean frog will result in a better sample. However, if the animal is dirty after capture do not wash it. This is suggested for three reasons: a) rinsing free zoospores off skin may reduce chances of detecting light infections. b) Washing off the animal may introduce large amounts of pathogen into the aquatic environment and facilitate transmission. 4 c) washing the animal in potentially contaminated water might introduce zoospores onto its surface, resulting in a false positive.