KyWW Training Module 4 provides guidance for bacteriological monitoring. Please view the video and take the completion quiz prior to attending an in-person training workshop.

Sampling Resources:

• R-Card Instructions

• One-Page "Cheat Sheet" for Field 

• One-Page "Cheat Sheet" for Incubation

• NEW Video Tutorial on using the pipette to collect stream water

• Practice R-Card Counting Exercise

• Data Form (for recording your results)

R-Card supplies will be provided by KYWW as funding allows. 

 
Where Support Hubs are established, the cards and pipettes will be available for pickup.  Otherwise, they will be mailed to samplers prior to each of the three annual sampling events.

Samplers also have the option to purchase supplies using the links to the left. If you purchase your own R-cards, please note the storage guidance from Roth Bioscience.

• R-CARDS© E. coli (3 cards/site needed)

• 1 mL pipette 

• Incubator

KYWW incubators for samples will be housed at Support Hubs, where available. Volunteers also have the option to purchase or make their own.

• Most egg incubators will work and can be purchased at farm supply stores. Watershed Watch is purchasing the Hova-Bator incubator.

• Homemade incubator also acceptable with proper temperature control (small cooler, light bulb, thermometer, e.g. 4W bulb in 1 cu. ft. cooler)

Instructions for DIY Incubator:  (SEE DETAILED INSTRUCTIONS ON LEFT)

A small cooler or other type of insulated container with a lid may be used as an incubator. Uniform temperatures may be maintained using a small light bulb, such as a night light attached to an extension cord. Use a piece of cardboard or plastic to shield the sample plates from direct light. A thermostat is not needed. The number and/or wattage of bulbs may be adjusted to maintain the proper temperatures, depending on the ambient room temperature and size of incubator. For example, a 4-watt bulb in a 1 cubic-foot container usually makes a good incubator. A thermometer is needed to determine ambient incubator temperature. This can be obtained from the AWW water chemistry test kit or a dial thermometer found in garden centers and hardware stores. The thermometer should be placed within the incubator away from the heat source or inserted through the lid so temperature may be read without opening the container. 

Some samplers will have the option to conduct lab analysis for E. coli when funding is available.  The procedure for sample collection in bottles will follow KYWW's Grab Sample Guidance.  Samplers will be given instructions for delivering water samples to the lab along with a Chain of Custody form for tracking the sample and reporting sample results to KYWW.

Chain of Custody Form for lab analysis

Instructions for Completing Chain of Custody Form

Grab Sample Guidance for E. coli:

• Enter the stream at a downstream location from your sampling point and walk upstream to avoid disturbing the stream bottom.

• Be sure to collect your sample at the same location each time in the middle of your concentrated flow location and at mid-depth of the stream. 

• Take caution not to contaminate samples with material from the bottom of the stream.

• Samples should not be collected when sample container cannot be submerged without fear of collecting sediment or other material from the stream bed. 

• Use caution when sampling in deeper water and watch out for deep pools. 

E. coli Grab Sample Procedure:

• When collecting an E. coli grab sample remember to put on disposable gloves to avoid any potential contamination. 

• Do not rinse out the bottle, since the bottles used for the collection of bacteria samples are sterilized. 

• Point the mouth of the bottle directly over the surface of the water, dip and scoop the bottle upstream beneath the water surface, and through the water column in one swift motion to fill above the line marked on the bottle.

• If a preservative tablet is in the bottle, alter the grab sample procedure and angle the mouth of the bottle so as not to spill the preservative while collecting the sample. 

• Cap the bottle, leaving at least a small airspace at the top to allow lab mixing. 

• Place in wet ice immediately.

Since the sample bottles are sterilized, if you do not fill your sample bottle above the marked line, it is not a usable sample. That sample bottle should be discarded.