Enterococci: Part I: Preparing Samples

Enterococci Testing

Preparing Samples

  1. Before leaving the lab, ensure you have a sterile IDEXX bottle for each sample site, labeled with the site name or number and collection date in permanent marker.
  2. At the site, leave the bottle sealed until it is submerged. Be sure to wear gloves, keep the collection bottle upstream from your hands, and to not disturb any sediment. Open the bottle underwater and allow to fill, then pour off a small amount of sample before capping, so the bottle is not completely full.
  3. Keep the sample in a cooler until back at the lab; if not preparing samples immediately, place the bottle in the fridge.
  4. In the lab, for each sample site, take out another sterile IDEXX bottle, an IDEXX tray, an Enterolert capsule, and a sterile pipette. Also take out a pipette bulb, a permanent marker, several bottles of autoclaved water and a pair of gloves for each person. Turn on the sealer, it cannot be used until the green light is illuminated.
  5. Label each IDEXX tray following the template; complete the data fields on the left half. Label each new, sterile IDEXX bottle with the site names or numbers. Ensure each person is wearing gloves.
  6. Take your first sample bottle and the respective sterile bottle. Shake the sample bottle, then using the sterile pipette, measure out 10 mL of sample water and pipette into the empty sterile bottle.
  7. Fill the bottle containing 10 mL of sample water with autoclaved water to the marked 100 mL line.
  8. Take one of the Enterolert capsules and empty into the mixture of sample/sterile water. Close the bottle and mix until the solid dissolves by slowly turning upside down then right side up and repeating.
  9. Once the solid is dissolved, pour the now yellow liquid into the corresponding IDEXX tray, then place the tray into the mold and run through the sealer. Remove tray from mold and set aside.
  10. Repeat steps 6-8 for each site.
  11. Place all sealed trays into the incubator, and leave for ~24 hours.