Answer to review questions

1. Explain why the following steps are essential during subculturing:

a. Flaming the inoculating instrument prior to and after each inoculation.
The importance of flaming of instrument is in order to reach sterility.
b. Holding the test tube caps in the hand as illustrated in Figure 1.1 on page 8.
In order to do not contaminate the caps or tubes because of possible contamination on hands.
c. Cooling the inoculating instrument prior to obtaining the inoculum.
If instrument is not cooled prior to the excess heat can kill the bacteria or fix the samples.
d. Flaming the neck of the tubes immediately after uncapping and before recapping.
In order to do not spread other bacteria on the neck of tube into nutrient, and also to do not
spread incubating bacteria through the neck
2. Describe the purposes of the subculturing procedure.
The main purposes are to separate the bacteria to make observations on them, and to prepare
stock cultures. Moreover, the above procedure should be done aseptically in order to avoid
contaminations.
3. Explain why a straight inoculating needle is used to inoculate an agar deep tube.
The reason for this is that inoculum has to be pulled out from the bottom of tube through a
straight line by line of insertion.
4. There is a lack of orange-red pigmentation in some of the growth on your agar slant labeled
S.marcescens. Does this necessarily indicate the presence of a contaminant? Explain.
Actually this could mean that there was wrong inoculation of bacteria or incorrect nutrient for
bacteria growth.
5. Upon observation of the nutrient agar slant culture, you strongly suspect that the culture is
contaminated. Outline the method you would follow to ascertain whether your suspicion is
justified.
To grow the nutrient agar slant culture more in order to observe the growth of unknown bacteria.

Introduction
Serratia marcescens is a short rod-shaped and facultative anaerobe bacterium classified as an
opportunistic pathogen. Firstly, the bacterium was discovered
Materials and methods
Results
Discussion
Conclusion