Answer : disinfectant test are usually performed using 5x5 cm tiles of surface material ,referred to as test carriers ,contaminated with relevant micro organism
The disinfectant is applied at a strength used in practice ,leftvon the test carrier for an appropriate time ,dropped into a neutralizing solution ,and rinsed .
The micro organism in the rinsing solution are then investigated ,either by incubating the solition on agar plates or the filter obtained after membrane filtration is incubated on agar.
This is followed by enumeration procedure .
Example :
Step 1 : suspension method
Step 2 : bacillus subtilis , escherichia coli ,candida albicans ,aspergillus niger ,staphylococcus aureus .
Step 3 : sop for preparation of microbiological media.
Suspension method :
Objective :
To establish the test concentration and the contact Time a suspension test estimate the invitro bacterial activity of disinfectant under microbial strain
Procedure :
Prepare the culture suspension from the original culture as per SOP for preparation of microbiological culture media .
Select the dilusion which will yield 10 '5 to 10 '6 cells per ml.
Prepare 10 ml of test dilution to be tested with sterile distilled water.
Vortex the tube for 1 minute .
Add 0.1 ml of any one culture containing 10'5 to 10'6 cells per ml into the test disinfectant with the dicided concentration
The final concentration shall be 10'4 to 10'5 cells per the tube .consider preoaration for 0 minute .
Prepare like same above for other 3 different sets for 3 different time intervals .
Give contact time 0, 5 min,10 min,15 minutes .
After specified contact time ,filter the samples through 0.45 m membrane filter.
Give three washing of 100 ml each with sterile peptone water /sterile water.
After filtration place it in soybean casein digest agar.
Incubate bacterial culture at 20 _25 degree centigrade for 72 to 120 hours .
Afterincubation count the number of colonies present on membrane .
Note down number of colonies .
Select plates which have least to nil counts .
Proceed in the same manner taking all the cultures to be tested .
Contact time for the usage of disinfectant will be set on basis of the results ,which will have least counts .
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