EXERICSE 11

SEPARATION OF PURE CULTURES FROM A MIXED SUSPENSION

INTRODUCTION
Bacteria grow among diverse and mixed populations, where it is impossible to study a
single species of bacteria. Therefore, it is necessary that the pure culture of bacterial
colonies be obtained for studying. Pure culture is defined as a mass or group of cells arising
from the same parent cell. Pure culture techniques, such as streak plate, isolate bacterial
colonies from mixtures so that colonies comprising of the identical organisms can be
studied. Separation of pure culture is vital for characterizing a single species of bacteria
otherwise presence of contaminants can lead to inaccurate observations.

AIM
To culture the microorganisms on artificial medium and observe the growth pattern and
colonial characteristics. Cultural methods not only help in identification but also are useful
in isolation and determination of kind and load of microorganisms present food stuff we
consume.

MATERIALS
1: A mixed suspension of micrococcus lutea and E or Coli
2: 1 Nutrient Agar plate
3: Gram Stain Reagents
4: Bunsen burner & Microbiological loop

METHODS
We Arranged all the supplies needed for the procedure on the laboratory bench and firstly
we placed the Bunsen burner to the right side on the bench and the agar plate on the left.
Light up the Bunsen burner and heat up the loop to sterilized. Move the loop, back and
forth many times across the agar surface from the rim to the center of the plate on a zigzag
motion. We repeat the process over and over to acquire subsequent manipulations.

RESULTS
This are the results we obtained after doing separation of pure culture from a mixed
cultures as we went through the above procedures.

DISCUSSION
By observing the results, it is visible that the streak plate technique isolates individual
bacterial cells (colony-forming unit) on the surface of an agar plate using a wire loop. The
streaking patterns shown in the figure above result in continuous dilution of the inoculum
to give well separated surface colonies. Once again, the idea is to obtain isolated colonies
after incubation of the plate. We haven’t encountered any problems in separating pure
culture from a mixed culture as we master the techniques.
In my personal point of view, I could say that, both colony and cell morphology are
characteristics of bacteria and which are also useful in the identification of an unknown
microorganisms. When a bacterium grows on a solid agar surface, the number of cell
increases until a visible mass of cell or colony appears, like what is shown in the result.

CONCLUSION
To conclude, the most commonly used method in the laboratory for
isolating microbes is the streaking plate and this method rely on dilution of bacterial cells
in a sample
to the point at which a single cell can divide giving rise to a single pure colony . The
pure colony is essentially a clone of cells that all are identical copies of the original
cell and can be used for further study. This lab practical encourages us to understand the
theory of bacterial cell morphology and cell division in correlation to cell structure.
REFERENCES

1. Azlin Sham Rambley (2018), Lecture Note, MLT 415, HS241, Chap. 9, Title: Media,
pg 1-38

2. Azlin Sham Rambley (2018), Lecture Note, MLT 415, HS241, Chap 6, Title:
Bacterial Physiology and Metabolisms, pg 1-25.

3. Divakaran A/L Chandrahasan (2012), Media Preparation, Isolation of Pure Culture

and Bacterial Growth, 1-6.

4. General Microbiology FT262 Laboratory Manual, Reviewed by R. Gubag- SIPOU.

July 2014. Page. (25-27)