Introduction

Microbiology can be defined as the study of microorganisms, namely bacteria.

Bacteria can manifest any known location on earth. In order to further the study of
microbiology and combat the problem of fast growth of bacteria, students need to
familiarise themselves with aseptic techniques. Aseptic techniques is a method used
to prevent microorganisms from growing in an area.
Aseptic techniques play an important role in the field of microbiology. By
using aseptic techniques, contamination can be prevented and accurate results
will be achieved. Microbiologists use aseptic techniques for different procedures
such as transferring cultures, inoculating media and isolation of pure cultures.
Aseptic techniques affect the results we achieve in any given practical.

Methods and Materials

Refer to Bacteriology (Micr213) Practical Manual. Practical 1(Page 9-18)
The pour plate technique was not carried out.

Results
Broth culture inoculation (E.coli) Bacteria sediment at the bottom of the test
tube. When the test tube was shaken, the broth culture became turbid.
Agar deep inoculation (Micrococcus luteus) Growth along the line of
inoculation with a mucoid growth above the agar in the test tube.
Agar slant inoculation (E.coli) Bacteria was white in colour and showed a
beaded spreading growth pattern with a watery mucoid growth.

Figure 1: Drawing showing the distribution of colonies on the streak plate

Discussion
E.coli showed growth at the bottom of the broth culture test tube, in the
presence of an oxygen deficit. The movement of test tube created turbidity in
the broth culture which shows that the bacteria is motile.
The agar deep inoculation of M.luteus showed growth along the line of
inoculation which means the bacteria is not motile.
The agar slant inoculation of E.coli showed a spreading growth pattern which
shows that bacteria is motile. Growth was shown where the oxygen
concentration was the highest.
The results show that M.luteus is a non-motile bacteria. The mucoid growth
on the top of the agar shows that M.luteus is an obligate aerobe since most
growth is shown where the oxygen concentration is the highest.
The results show that E.coli is a facultative aerobe, since inoculations showed
growth in both high and low concentrations of oxygen. E.coli is a motile
bacteria, shown by the spreading growth pattern.
Air contamination is prevented by sterilizing the inoculation loop over the Bunsen
burner. The back and forth inoculation technique is avoided; the streak is carried out
with a medium slant in a straight line so that the growth pattern of the bacteria can
be determined.
By gaining knowledge of the characteristics and morphology of different types
of bacteria, It will be easier to identify in future microbiology practicals.

Conclusion
The aim of learning aseptic techniques, carrying out inoculations and
identifying microbial growth patterns has been satisfied. Aseptic techniques
ensure that no contaminants enter the culture, ensures that the bacteria does not
come into contact with the user and also ensures that the bacteria does not spread
to the outside environment. Inoculating loops and needles are important tools that
prevent direct contact with bacteria and prevent students from handling and
transferring bacteria by hand. Obtaining pure cultures, cultures of a single strain,
enable viewers to distinguish distinct characteristics and differences between
bacteria.
Bacteria is something that we cant live with, but also cant live without.
Therefore it is important for people to understand why it is beneficial to us,
and even more important to understand how harmful it can be. With these
results we have a deeper understanding of ubiquitous bacteria, their
morphology and factors that favour their growth.

References
Bacteriology(Micr213) Practical Manual 2015
http://en.wikipedia.org/wiki/Facultative_anaerobic_organism
http://amrita.vlab.co.in/?sub=3&brch=73&sim=212&cnt=1
http://www.scienceprofonline.com/microbiology/bacterial-colony-morphologyidentification-unknown-bacteria.html
Willy, Sherwood and Woolverton. Prescotts Microbiology. 9th edition.

Aseptic Techniques/Transfer and Isolation of

Microorganisms