Answers to exam of March 8, 2011

Multiple choice answers are on the sheets returned to each student. Question 18 had two correct answers and
credit was given for either a or d.

Short answer questions:

76. Why was the development of pure culture techniques necessary for the application of Koch’s
Postulates?

The essential purpose of Koch’s postulates was to show that a specific organisms caused a specific
disease. The only way to know that for sure is to have a pure culture of the microorganism in
question and show that it caused the disease when inoculated into a healthy animal. Without pure
culture techniques, he would probably be doing the test inoculations with mixed cultures and it would
then be impossible to know which one was causing the disease.

Descriptions of action of agar or its properties were not part of this question, since it did not ask
HOW the pure cultures were created. Similarly a description of the steps in Koch’s postulates
without explanation such as that above does not answer the point of this question.

77. Why do transmission electron microscopes have better resolution than bright field microscopes?

The primary difference in resolution between the two instruments is the wavelength of the
electromagnetic radiation used for illuminating the specimen. The bright field microscope uses
visible light and the transmission electron microscope (TEM) uses beams of electrons which behave
as a wave of very short wavelength,

To get full credit, it was necessary to explain how the shorter wavelength improves resolution, not
just say it. The easiest way to do this was to provide the equation for resolution and discuss that in
this context, although that was not required. Several students mentioned index of refraction and this
is a physical property that has no meaning in the light microscope. Other students mentioned the
electromagnets used to focus the electron beam in the TEM. Although this is an accurate statement
about focusing the TEM, this has nothing to do with resolution.

78. Yeast cells, which are eucaryotic, stain Gram positive. Explain.

The Gram stain procedure provides its differential results entirely because of the thickness of the cell
wall. The chemical composition of the wall has no effect on the staining reaction; it is just the
physical matter of thickness. Yeast cell walls do not contain peptidoglycan, but since they are thick
they dehydrate upon the addition of alcohol and collapse.

Common mistakes included references to membranes instead of cell walls and some students said
that yeast cells have peptidoglycan, which is a very serious mistake.
79. Why is lysozyme active against all bacteria and not just against actively growing bacteria?.

Lysozyme cleaves the polysaccharide chains in peptidoglycan whether the cells are actively growing
or not. This is different from penicillin, which interferes with the formation of peptide crosslinks, but
only in actively growing cells.

Common wrong answers were to say that lysozyne cleaves peptide linkages or to imply that it was
active against both polysaccharides and peptides.

80. Consider two different bacteria. One is in a solution that has no attractants or repellents and the other
is in a solution with a uniformly high concentration of an attractant such as glucose. Which
bacterium will have longer runs and less frequent twiddles? Explain.

The two cells behave the same, displaying short runs and frequent twiddles, The key concept to keep
in mind for this question is the need for there to be a gradient in order for the bacteria to display
chemotaxis. The two cells referred to in this question are both in solutions with no gradient. Once
has high nutrients (glucose), but since there is no gradient, then the cells behave just as though there
were no attractants.