6. Test Procedure6. Test Procedure

6. Test Procedure

Perform the following test using each of the four test organisms (3.1) except where the Standard directs otherwise. It is not necessary to test with all organisms simultaneously.
6.1 Add 3ml of diluted disinfectant to a capped glass container.
6.2 Start a timing device. Immediately inoculate disinfectant with 1ml of culture (prepared in 3.2) and mix by swirling.
6.3 At 8 minutes, subculture one drop (0.02ml + .002ml) into each of 5 tubes containing recovery broth. To ensure delivery of 0.02ml into the first tube of recovery broth at exactly 8 minutes, it will be necessary to withdraw a suitable amount from the disinfectant test mix shortly beforehand. This must be immediately preceded by vortexing. Surplus sample must be returned to the test mix. (See Note D).
6.4 Except where prescribed, at 10 minutes, inoculate disinfectant with a further 1ml of culture, and mix by vortexing.
6.5 Except where prescribed, at 18 minutes, proceed as in 6.3.
6.6 Mix the contents of all tubes of recovery broth by vortexing. Incubate at 370 + 10C for 48 + 2 hours.
6.7 Examine for growth and record results.
6.8 For each test organism repeat steps 6.1‑6.7 on each of 2 subsequent days, using a fresh disinfectant dilution and a freshly prepared bacterial suspension.