Environment Monitoring of Microbiology Section
This Document describes (Standard Operating procedure) SOP for Environment Monitoring of Microbiology Section
SOP for Environment Monitoring of Microbiology Section
I. Purpose & Scope:
- The purpose of this SOP is to lay down the procedure for Environment Monitoring of Microbiology Section
- This Standard operating procedure shall be applicable for Environment Monitoring of Microbiology Section
II. Responsibilities:
- All Quality Control and Quality Assurance personnel shall be responsible to follow and implement this SOP of for Environment Monitoring of Microbiology Section.
III. Introduction and Procedure:
Procedural part for environment Monitoring of Microbiology Section:
Preparation of Plates:
- Prepare Soybean Casein Digest Agar (SCDA) plates for monitoring of Total Viable Aerobic Count as per current version of respective SOP.
- To avoid the dryness in plates due to exposure, add 1% Glycerin in prepared media before sterilization.
Environment monitoring of microbiology testing area by settle plate Method:
- For monitoring of Microbiology testing area perform the ‘Entry/Exit procedure’ as per respective SOP.
- Expose the SCDA plates, at specified locations indicated in the Annexure-I.
- Label individual plates with the media lot number, Name of location/ Plate I.D. and date of exposure.
- Place these plates at the designated locations on the SS stands provided for settle plate exposure. Before exposure of plates, clean the SS stands with 70%IPA.
- Carefully open the lid of the plates and place the lids on the stand adjacent to the plate. Expose the plates for two hours. After exposure, collect the plates and incubate them in the inverted position.
- Perform the dual temperature incubation for Settle plates.
- At first incubate the plates in incubator at 20 to 25°C for 72 hours, observe the plates and record the count in Annexure-II. Then transfer and incubate the plates in incubator at 30 to 35°C for 48 hours.
- The number of colony forming units (CFU) observed on the plate after each incubation period shall be recorded. Total Viable Aerobic Count (TVAC) is the summation of the TBC and TFC after incubation period of 5 days. The count shall be expressed as ‘CFU / Plate’. Record the results in Annexure-II.
Limits and Frequencies for Microbiology Testing Area:
| Activity | Grade | Alert limit | Action limit | Maximum Limit
(cfu/plate) |
Frequency |
| Total Bacterial Count | D | 50 cfu/ plate | 80 cfu/ plate | NMT 100 cfu/plate | Once in Fortnightly |
| Total Fungal Count | D | 03 cfu/ plate | 04 cfu/ plate | NMT 05
cfu/plate |
- Positive control: For positive control, Refer the media fertility test performed during Media preparation.
- Negative Control: Keep one plate of the same media lot as negative control with test plates. Keep it unexposed and incubate it along with test plates.
Preparation of Trends for Environment Monitoring of Microbiology Section :
- Quarterly: Prepare the routine trends once in Quarterly.
- Yearly: Also prepare the yearly trends. For preparation of yearly trend, take the month period from January to December and maximum count of respective month.
Environment Monitoring of Laminar Air Flow by settle Plate method :
- Perform the Environment monitoring of laminar air flow (LAF) of MLT Room and Culture Handling Room by Settle Plate Method.
- Perform the ‘Entry/Exit Procedure’ as per respective SOP.
- Ensure that LAF is clean. Check the differential pressure by manometer reading, it should be between 7.5 to 13 mm of water column.
- Expose the SCDA plates at the specified locations in LAF, indicated in Annexure-I.
- Label individual plates with the Media lot number, Name of location/ Plate I.D, Instrument I.D and date of exposure.
- Carefully open the lid of the plate and place it, adjacent to the plate. Expose the plates for four hours. After exposure, close and collect the plates. Incubate the plates in inverted position.
- Perform the dual temperature incubation for Settle plates. At first incubate the plates at 20 to 25°C for 72 hours, in incubator. Observe the plates and record the count in Annexure-III. Then transfer and incubate the plates in incubator at 30 to 35°C for 48 hours.
- Count the number of colony forming unit (CFU) observed on the plate after each incubation period. Total Viable Aerobic Count (TVAC) is the summation of the TBC & TFC after incubation period of 5 days. The count shall be expressed as CFU / Record the results in Annexure-III.
- Positive control: For positive control, Refer the media fertility test performed during Media preparation.
- Negative Control: Keep one plate of the same media lot as negative control with test plates. Keep it unexposed and incubate it along with test plates.
Limit and Frequency for Monitoring of Laminar Air Flow :
| Activity | Grade | Alert limit | Action limit | Maximum Limit
(cfu/plate) |
Frequency |
| Total Bacterial Count | A | – | – |
< 1cfu/plate
|
Once in a week |
| Total Fungal Count | A | – | – |
< 1cfu/plate
|
Actions to be taken in case of microbial counts exceed the Limit:
- For details of action to be taken in case of Microbial count exceeds the limit.
IV. Annexures:
Annexure I: List of location for Environment monitoring of Microbiology section by Settle plate method.
Annexure II: Environment monitoring report of Microbiological testing area by Settle plate method.
Annexure III: Environment monitoring report of Laminar Air Flow by Settle plate method.
Annexure I
List of location for Environment monitoring of Microbiology section by Settle plate method
Page No: 1 of 1
- A) MICROBIOLOGY SECTION (MICROBIOLOGY TESTING AREA AND CULTURE HANDLING AREA):
| Plate I.D | Name of Location
(Plate Exposed for 2 hours) |
Grade | Maximum Limit (cfu/Plate) |
| D | NMT100cfu/plate | ||
| D | NMT100cfu/plate | ||
| D | NMT100cfu/plate | ||
| D | NMT100cfu/plate | ||
| D | NMT100cfu/plate | ||
| D | NMT100cfu/plate |
- B) MONITORING OF LAMINAR AIR FLOW:
| Plate I.D | Name of Location
(Plate Exposed for 4 hours) |
Grade | Maximum Limit (cfu/Plate) |
| A | < 1cfu/plate | ||
| A | < 1cfu/plate | ||
| A | < 1cfu/plate |
Annexure II
Environment monitoring report of Microbiological testing area by Settle plate method.
Page No: 1 of 1
| Date of Sampling | Done by | ||
| Time of Exposure | Frequency | ||
| Status | Static / Dynamic | Media Lot No. | |
| Date of Completion | Observed by |
| Plate I.D. |
Location (Plate exposed for 2 hours at following Location)
|
Grade | Incubation condition:
(20-250C/72 hours) Date:——————— Incubator I.D.:———– Observed On:———— |
Incubation condition:
(30 -350C/48 hours) Date:——————– Incubator I.D.:———— Observed On:————- |
||
| Total Viable aerobic Count (cfu/plate) | ||||||
| Bacteria
(cfu/Plate) |
Fungal
(cfu/Plate) |
Bacteria
(cfu/Plate) |
Fungal
(cfu/Plate) |
|||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| Activity | Grade | Alert Limit | Action Limit | Maximum Limit |
| Total Bacterial Count | D | 50cfu/plate | 80cfu/plate | NMT100cfu/plate |
| Total Fungal Count | D | 3cfu/ plate | 4cfu/plate | NMT5cfu/plate |
Positive Control:——————————- Negative Control: —————————-
Conclusion: Total viable aerobic count of Environment monitoring by Settle plate method are found Satisfactory/Not Satisfactory.
Observed By: Checked By:
Date: Date:
Annexure III
Environment monitoring report of Laminar Air Flow by Settle plate method
Instrument I.D.: _____________
| Date of Sampling | Done by | ||
| Time of Exposure | Frequency | ||
| Status | Static / Dynamic | Media Lot No. | |
| Date of Completion | Observed by |
| Plate I.D. |
Location (Plate exposed for 4 hours at following Location)
|
Grade | Incubation condition:
(20-250C/72 hours) Date:——————— Incubator I.D.:————- Observed On:————— |
Incubation condition:
(30 -350C/48 hours) Date:——————– Incubator I.D.:———— Observed On:————- |
||
| Total Viable aerobic Count (cfu/plate) | ||||||
| Bacteria
(cfu/Plate) |
Fungal
(cfu/Plate) |
Bacteria
(cfu/Plate) |
Fungal
(cfu/Plate) |
|||
| A | ||||||
| A | ||||||
| A | ||||||
| Activity | Grade | Alert Limit | Action Limit | Maximum Limit |
| Total Bacterial Count | A | — | — | <1 cfu/plate |
| Total Fungal Count | A | — | — | <1 cfu/plate |
Positive Control:——————————- Negative Control: —————————-
Conclusion: The Total viable aerobic count of Environment monitoring by Settle plate method are found Satisfactory/Not Satisfactory.
Observed By: Checked By:
Date: Date:
