SOP For Microbial Monitoring of Drainage Water

SOP For Microbial Monitoring of Drainage Water is described in this post which you can follow in the section of the microbiological lab.

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Objective:

This procedure has been established for sampling and testing of drain water before and after sanitization process in the pharmaceutical manufacturing organization.

Scope:

The scope of this procedure is applicable to the sterile manufacturing & testing area i.e.(Microbiology Lab).

Responsibility:

Microbiologist.

Sterile Area Pharmacist.

Microbiology Lab Attendant.

Accountability:

Quality Control Manager.

Procedure:

Sampling:

Sample approximately 20ml of drain water (Before and after sanitization) in a sterile beaker individually from each drain of the plant with the help of a sterile pipette.

Collect the sample by using pipette filler for the collecting drain water for microbiological testing.

Testing For Total Viable Count:

For making First dilution:(1:10 dilution): Take 1ml of drain water as sample in 10ml of sterile Peptone water/ 0.9% normal saline solution.

For making second dilution: (1:100 dilution): Take 1ml of drain water as sample in 100ml of sterile peptone water/ 0.9% normal saline solution.

For making third dilution: (1:1000 dilution) Take 1ml of drain water in 1000ml of the sterile peptone water/ 0.9% normal saline solution.

After preparing the three dilutions of the drain water transfer aseptically 1ml of the sample from each dilution to the three different sterilized Petri plates

Add about 20ml-25ml of the sterilized molten soybean casein digest agar for bacterial growth and sabouraud dextrose agar for yeast mold growth.

After adding the culture media, mix the sample properly and allow to solidify.

Before setting for incubation mark the plates with the location No. and date on each plate.

Incubate the SCD agar plate at 30°-35°C for bacteria for 2-3 days, and Sabouraud dextrose agar plate at 20°-25°C for fungi for 5 days.

After the completion of incubation period, observe the plates for growth in terms of number of CFU (colony forming unit).

Calculate the average CFU of two plates in terms of cfu/ml.

Microbial Limit Test for (E.coli & Salmonella)

For making (1:1 dilution) Take 10ml of drain water and add in 10ml of sterile Peptone water/ 0.9% normal saline solution.

For making (1:10 dilution) Take 10ml of drain water and add in 100ml of sterile Peptone water/ 0.9% normal saline solution.

For making (1:100 dilution) Take 10ml of drain water and add in 1000ml sterile Peptone water/ 0.9% normal saline solution.

After preparing the three dilutions of the drain water transfer aseptically 1ml of of the sample from each dilution to the three different sterilized petri plates.

and add about 20ml-25ml of the sterilized bismuth sulfite agar for salmonella and Macconkey agar for E.coli.

After adding the culture media mix the sample properly and allow to solidify.

Before setting for incubation mark the plates with the location No. and date on each plate.

Incubate the bismuth sulfite agar plate at 37°C for Salmonella and Macconkey agar for 37°C for E.coli for 24-48hrs

After the completion of incubation period, observe the plates for growth in terms of number of CFU (colony forming unit).

Calculate the average CFU of two plates in terms of cfu/ml.

Calculation

Formula For Calculation

CFU/ml= Number of CFU Observed × (ml)Dilution Used

For 1:10 dilution=N×10

For 1:100 dilution=N×100

For 1:1000 dilution= N×1000