CADIMAS, Mae Lailani P. Mr.

Andrew Montecillo
MCB 150 CD-2L December 9, 2009

EXERCISE 2
Soil as an Environment

OBJECTIVES
1. To differentiate the four types of soil – acidic, non acidic, waterlogged and
non-waterlogged based on physical characteristics such as pH and
moisture content;
2. To determine the abundance of different microorganisms, such as
bacteria, fungi, actinomycetes and algae, in samples of the three types of
soil;
3. To relate the physical factors in soil with the distribution and kind of
microorganisms present in the four soil types; and
4. To determine microbial activity in the three soil samples based on carbon
dioxide evolution.

METHODOLOGY

I. Kind and Distribution of Microorganisms in Soil

A.

B.
 C.

II. Carbon Dioxide Evolution

RESULTS

I. Kind and Distribution of Microorganisms in Soil

Table 1. Physical characteristics of the four soil samples.

Soil Type pH Initial weight Oven dry % Moisture
(g) weight (g) Content

Acidic Soil 5.9 20 15.49 29.12

Non Acidic Soil 5.4 10 8.72 14.68

Water-logged 6.9 10 8.08 23.76

Non water- 7 10 7.60 31.58

logged

Table 2. Abundance of the different microorganisms in three types of soil as

determined by the standard plate count and most probable number technique
and expressed in terms of colony-forming units per gram (CFU/g) of sample and
MPN, respectively.
Soil Type Fungi Bacteria Actinomycetes Algae
(CFU/g) (CFU/g) (CFu/g) (MPN)

Acidic Soil 2.4 x 106 3.6 x 107 3.1 x 106 99

Non Acidic Soil 1.0 x 106 1.9 x 106 1.4 x 106 50

Water-logged 1.0 x 106 9.6 x 105 7.6 x 105 77

Non water- 1.5 x 106 2.5 x 10-4 5.8 x 106 38

logged ESPC
 Table 3. Amount of carbon dioxide evolved in the four soil types.
Soil Type Data CO2 evolved
(mg/100g soil)
With peptone Without peptone

Acidic Soil 0.1 ml of 0.05N Pink disappeared ~0.11

HCl rapidly

Non Acidic Soil No pink 1.55 ml of 0.05N HCl 1.71

Water-logged No pink Pink disappeared Cannot be

rapidly determined

Non water- No pink 0.89 ml of 0.05N HCl 0.98

logged

ANSWERS TO QUESTIONS

1. Why is it ok to use non-sterile materials for soil samples?

Since the soil is a known habitat for various microorganisms, the
microorganisms present in the non sterile material used in sampling and
experiment is far outnumbered. However, it might be difficult to determine which
among the microorganisms came from the soil and which came from the
materials used.

2. Why is MPN used to quantify soil algae and not the standard plate count method?
The standard plate count is not recommended because algal colonies are
not distinguished from each other since algae are filamentous microorganisms
that can form dense networks.

3. What are some potential errors associated with dilution and plating?

The potential errors associated with dilution and plating are: (1) the
number of dilutions may not be enough to reduce the number of microorganisms
to be observed therefore the growth during plating may be abundant (2) isolation
of all microorganisms present may not be possible since they do not have all the
nutrients necessary for the growth of all organisms in the soil sample.

4. Why is it important to determine the dry weight of the soil samples used in the
experiment?
The dry weight may determine the exact number of organisms per mass of
soil. The presence of moisture in the soil may be accounted for the abundance of
microorganisms in a certain area.

5. How does soil moisture content affect the activity of aerobic and anaerobic soil
microorganisms?
When the soil has more moisture, the growth of anaerobic soil
microorganisms is favored since the water in the soil occupies the pores which
 are previously occupied by air, therefore limiting the diffusion of air, particularly
oxygen resulting in an anaerobic condition.

6. How does soil moisture affect transport of soluble pollutants?

In soil, movement of water and moisture is observed. As the water activity of
the soil changes, it carries the soluble pollutants that are usually dissolved in the
water present in the soil. The dissolved pollutant in the soil moisture may be
included in the change in physical state of the water (e.g. evaporation) or may be
transported to other places through other water activities.

REFERENCES

Atlas, R.M. and R. Bartha. 1993. Microbial Ecology: Fundamentals and Applications
3rd ed. California: The Benjamin/Cummings Publishing Co., Inc.

Madigan, M., J. Martinko, and J. Parker. 2003. Brock Microbiology of Microorganisms

10th ed. Prentice Hall, New Jersey.

Prescott, Lansing M. 1993. Microbiology 2nd ed. Wm C. Brown Publishers, Iowa.