ANALYSIS OF AIR

Aim:
To study the air flora of any environment by gravity sedimentation method

Principle:
Microbial flora of air is transient and variable. Organisms usually adhere onto
particulate material such as dust, carbon, saliva etc. The conditions of air
prevent growth of organisms vegetative cells like Staphylococci, Streptococci
and Mycobacteria resist drying and are carried over long distances

Theory:
Air sampling is done by impinging air onto a solid surface during a brief
exposure. The agar surface subsequently is incubated till visible colonies
appear. The sampling method rests on sedimentation of particulate matter
under the influence of gravity.

Materials Required:
LB Agar plates
Potato dextrose Agar plates

Procedure:
Expose the LB agar and Potato Dextrose Agar plates to air for 10 min/1hr and
then incubate them at RT and 37oC for 48 hrs.

Observations:
Location

LB Agar

PDA

Car Park
M. Sc Part I
Lab
Laminar

28oC
140
90

37oC
125
115

28oC
25
20

15

12

Calculations:
The rate of micro-organisms (biological aerosols) falling on to a critical
surface can be calculated by the following formula:
c= Vc X Pc,
where c is the contamination flow, i.e., the count of settling cfu per unit
surface and per unit time; Pc is the contamination density, i.e., the count of
cfu per unit volume; Vc is the contamination velocity, i.e., the settling
velocity of cfu.

Result:
Conclusion:
The qualitative and quantitative analysis of air was done the resulting
colonies were observed and studied.

ANALYSIS OF SOIL
Aim:
To carry microbial analysis of the given soil sample

Principle/Theory:
The microbial analysis of soil is carried out to find out the different types of
microbes present in the soil and their effect on the soil. Microbes in the soil
are important because they affect the structure and fertility of the soil. Soil
microbes can be classified as bacteria actinomycetes fungi algae and
protozoa.

Material Required:
Glassware:
Sterile Petriplates
Sterile 1 ml and 10 ml pipettes
Sterile test tubes
Sterile beaker
Spreaders
Media:
Luria Bertani Agar (LB agar)
Potato Dextrose Agar (PDA)
Physiological saline (0.85%)
Sample:
Soil from university compound

Procedure

1. Weigh 1 gm of the soil sample and resuspend in 10 ml of sterile distilled

water.
2. Prepare serial dilutions of the sample.
3. Plate out 100 l of 10-6, 10-7 and 10-8 on LB agar plates (two each) and
the lowest one on PDA.
4. For pour plate, add 100 l of the above dilutions to 25 ml of the molten
medium (maintained at 50oC).
5. Incubate one plate of each dilution at 37oC and the other plate at 28oC
for 48 hours. Incubate PDA at 28oC for 48 hours.
6. Check for colonies.
7. Quantify using cfu/ml

Observation:
Dilution
10-6
10-7
10-8

LB Agar
28oC
700
75
8

37oC
670
65
10

PDA
28oC
NA
10
2

Figure 1: Soil sample on PDA plates

Calculation:

Result:
Conclusion:
The soil sample was analyzed and various organism found were Isolated
and studied.

ANALYSIS OF WATER
Aim:
To determine the microbial load in the given sample of water

Principle/Theory:
There are different organisms present in water some are pathogenic some
are not. The pathogenic bacteria are generally gram negative short rods
called coliforms which are found in water depending if the water was
stagnant or polluted a number of other organisms such as algae n different
microbes can be found in it. By performing the water analysis we can find out
the different organisms present and can also isolate a possible threat of
contamination.

Material Required:
Glassware:
Sterile Petriplates
Sterile 1 ml and 10 ml pipettes
Sterile test tubes
Sterile beaker
Spreaders
Media:
Luria Bertani Agar (LB agar)
Potato Dextrose Agar (PDA)
MacConkeyss Agar
Physiological saline (0.85%)
Sample:
Soil from university compound

Procedure
1. Collect water sample from pond and water cooler.
2. Perform serial dilutions of both the samples.
3. Plate out 100 l of 10 -6, 10-7 and 10-8 dilutions of pond water sample on
LB agar plates (two each).
4. For pour plate, add 100 l of the above dilutions to 25 ml of the molten
medium (maintained at 50oC).
5. Plate out 100 l of 10-2, 10-4 and 10-6 dilutions of cooler water sample
on LB agar plates.
6. Incubate one plate of each dilution at 37oC and the other plate at 28oC
for 48 hours.
7. For MacConkeys and PDA plates, plate out 100 l of 10 -2, 10-4 dilution
of cooler water and 100 l of 10-4, 10-6 dilution of pond water.
8. Incubate MacConkeys plates at 37oC and PDA at 28oC.
9. Check for colonies.
10.
Quantify using cfu/ml

Observation:
Pond water count
Dilution

10-6
10-7
10-8

LB Agar
28oC
700
75
8

PDA
37oC
670
65
10

28oC
10
1
NA

MacConkey
s Agar
37oC
78
8
NA

Cooler water count

Dilution

10-2
10-4
10-6

LB Agar
28oC
133
20
4

PDA
37oC
120
11
1

28oC
5
NA

MacConkey
s Agar
37oC
70
8
NA

Figure 2: Cooler water Sample on LB Agar plate

Calculation:
Result:
Conclusion:
The analysis of water was carried out and the colonies of different microbes
were observed