1. Pharmaceutical Microbiology

2. Microscopic Examination of Microorganisms
Practical Part
Microscopic Examination of Microorganisms
Experiments
Identification of MOs
Different Staining Techniques

3. Experimental Microbiology

4. Pour Plate Technique Objectives :
To learn the pour-plate technique for using in other experiments such as streaking, sensitivity test and for MIC determination.

5. Materials : Procedure:- Tube of nutrient agar. Sterile Petri dish.
Continue
Materials :
Tube of nutrient agar.
Sterile Petri dish.
Bunsen burner flame.
Procedure:-

6. Place a tube of sterile nutrient agar in a boiling water bath
Place a tube of sterile nutrient agar in a boiling water bath. Keep the water level at the halfway mark.
(1)

7. Continue
(2)
When the agar is liquefied, remove the tube and allow it to cool to about 50°C.
Place an empty sterile Petri dish before you, top side up.

8. Pick up the tube of melted, cool agar, remove its closure.
Flame the mouth of the open tube.
(3)

9. With your free hand, remove the top of the Petri dish and quickly pour the agar into the dish.
(4)

10. Continue
(5) Replace the Petri dish cover Gently rock the closed dish, or rotate it in circular fashion on the bench top, being careful not to allow the still melted agar to wave up over the edge of the bottom half or onto the cover.

11. Continue
(6) Let the agar solidify without further disturbance. When it is quite firm, invert the plate.

12. Experiment 1

13. Distribution of Microorganisms in the Environment
Objectives :
To demonstrate the wide distribution of microorganisms in the air, on the skin, and in the air droplet produced by coughing in order to avoid sources of contamination.

14. Materials : Procedure: (As previous) Six nutrient agar tubes.
Continue
Materials :
Six nutrient agar tubes.
Six sterile Petri dishes.
Procedure:
(As previous)

15. Source of Contamination
No. of colonies of
Source of Contamination
Plate No.
Actinomycetes
Fungi
Bacteria
Air 1
Forced air 2
Cough 3
skin 4
Tap water 5
None 6

16. Continue
Incubate all plates at room temperature for hr.
After incubation, examine the plates for the presence of bacterial or fungal colonies or actinomycetes and describe each.

18. Source of Contamination
Results of Experiment:
No. of colonies of
Source of Contamination
Plate No.
Actinomycetes
Fungi
Bacteria
Air 1
Forced air 2
Cough 3
skin 4
Tap water 5
None 6

19. **Description of colonies from plate number (….)
Shape

20. Edge

21. Elevation

22. Colony description
Isolate 1
Isolate 2
Isolate 3
Shape
Color
Size
Edge (margin)
Elevation
Texture
Surface
Optical Character

24. Colony description Isolate 1 Isolate 2 Isolate 3 Shape Round , oval
Color
Yellow
Size
Medium to large
Edge (margin)
Entire
Elevation
Convex
Texture
Smooth
Surface
Optical Character
Opaque

25. Experiment 2

26. Techniques for Isolation of Pure Cultures
Objectives :
To perform the spread plate and/or streak plate inoculation procedure for the separation of a mixed culture so that discrete colonies can be isolated.

27. Materials : Tube of nutrient agar (10 ml). Sterile Petri dish.
Continue
Materials :
Tube of nutrient agar (10 ml).
Sterile Petri dish.
Wire inoculating loop.
Tube containing mixed culture of two microorganisms.
Permanent marker.

28. Continue
Procedure:
Invert the dish and by means of marker pen draw 5 groups, each with 3 parallel lines that intersect with each other.

30. Cool the agar to 50 ○C before pouring to avoid water condensation.
Continue
Using pour plate technique, aseptically pour the melted nutrient agar into the plate and leave to solidify.
N.B
Cool the agar to 50 ○C before pouring to avoid water condensation.

31. Continue
After solidification, invert the plate to avoid water condensation on the surface of agar.

32. Gently Mix the Wasserman containing mixed culture.

33. Flame the wire loop.

38. Place a loopful of culture on the agar surface at the beginning of group 1.

39. Re-flame the loop between each group

44. Results of Experiment:
Colony description
Colony 1
Colony 2
Colony 3
Shape
Color
Size
Edge (margin)
Elevation
Texture
Surface
Optical Character