TITLE: Field protocol for river water sampling 
AUTHORS: Christopher LeBoa, Sneha Shrestha

[DESCRIPTION]
This protocol is for the collection of water samples for molecular analysis. It was used for collection of river water samples from Kathmandu for detection of S. Typhi and Paratyphi A


[GUIDELINES]
NOTE: Bags are cleaned every day with 0.5% Hypochlorite followed by 70% ethanol before refilling the materials.

[STEPS]
SECTION: Bag Preparation for sampling (Two separate bags, one for samples and another for sampling materials Or a bag with separate compartments for samples and the materials)
1. Wipe the inner part of the sample bag with 0.5% Hypochlorite followed by 70% ethanol.
SECTION: Bag Preparation for sampling (Two separate bags, one for samples and another for sampling materials Or a bag with separate compartments for samples and the materials)
2. Place ice packs and a tube holder above the ice packs to place the collected samples.
SECTION: Bag Preparation for sampling (Two separate bags, one for samples and another for sampling materials Or a bag with separate compartments for samples and the materials)
3. Place the negative control tube.
SECTION: Bag Preparation for sampling (Two separate bags, one for samples and another for sampling materials Or a bag with separate compartments for samples and the materials)
4. Place each material in a separate clean ziplock bag before placing it inside the bag.
SECTION: Bag Preparation for sampling (Two separate bags, one for samples and another for sampling materials Or a bag with separate compartments for samples and the materials)
5. Always place the pH meter in upright position so as not to spill the KCl (where the electrode is dipped).
SECTION: Bag Preparation for sampling (Two separate bags, one for samples and another for sampling materials Or a bag with separate compartments for samples and the materials)
6. Refill the materials required each sampling day.
SECTION: Bag Preparation for sampling (Two separate bags, one for samples and another for sampling materials Or a bag with separate compartments for samples and the materials)
7. Replace discard bags each sampling day.
SECTION: Sample Collection
8. Use the ArcGIS collector app to get to the designated sampling points.
SECTION: Sample Collection
9. Collect 50ml of flowing river water sample in a sterile 50ml falcon tube by grab sampling technique.
SECTION: Sample Collection
10. Disinfect outer surface of the tube with cotton soaked in 0.5% sodium hypochlorite followed by 70% ethanol.
SECTION: Sample Collection
11. Let the surface dry and label the tube with a fine tip permanent marker. Label each sample with Collection site code and Collection date.
SECTION: Sample Collection
12. Eg: For Bagmati river site 1 sample collected on 13th March,2020 label the tube as
SECTION: Sample Collection
13. BAG-1 13/03/2020
SECTION: Sample Collection
14. Transfer into a backpack containing ice packs for transportation to the laboratory under   cold conditions.
SECTION: Sample Collection
15. Collect measurements of pH, ORP and temperature of the river water at the sampling point using a pH meter. After measurement at each site, rinse the probe of the pH meter with distilled water.
SECTION: Sample Collection
16. Record the results and complete the questionnaire in RedCap on a tablet.