MIC 205: Exam 1 Study Guide

Introduction to Microbiology

 Know what the Spontaneous Generation Debate was, which scientists played a part,
and how each contributed to settling the debate. Who finally disproved it? How?
Why didn’t the other experiments prove/disprove spontaneous generation?

Spontaneous generation is a theory that life can generate from non-living sources.
Scientists:
Francesco Redi: Did not believe in spontaneous generation. He made an experiment to
disprove it in 1668. Experiment consisted of meat in a flask. First flask was uncovered,
second had a film, and third had a cork. He found flies and maggots on the uncovered flask.
Robert Hooke: Observed microorganisms for the first time with a microscope and
discovered the “cell.”
Anton Van Leeuwenhoek: Wrote to the royal society of London for improving natural
knowledge about his observations on the plaque between his teeth. Rekindled spontaneous
generation debate after discovering microbes randomly appearing in rain water after a few
days.
John Needham: Conducted an experiment that “proved” spontaneous generation. He briefly
was selected as a member of the Royal Society due to this experiment. Experimented with
briefly boiling broth in a flask without sealing it for days, which was a flaw in the
experiment.
Lazzaro Spallanzani : Disproved the theory of spontaneous generation by recreating John
Needham’s experiment, however, making sure that the sample had not been exposed to air.

 Understand the Germ Theory of Disease, what steps were taken (or proposed) early
on to prevent infections?
Germ Theory was coined by Louis Pasteur in 1861. He said that the infection could be
prevented by pasteurizing products such as beer, wine, and milk. The Germ Theory states
that, “Many diseases are caused by organisms. These small organisms, too small to see
without magnification, invade humans, animals, and other living hosts. Their growth and
reproduction within their hosts can cause a disease.”

 Be able to explain how Koch’s Postulates are used to determine etiology of a disease.
Be able to determine how each postulate can be invalidated (like the slide in class).

Specific microbe is always associated with a given disease (not always true, multiple
organisms can cause some disease/symptoms. Not always caused by microbe: prison, fungus,
plaque, virus, etc.) Microorganism can be isolated from the diseased animal and grown in
pure culture (not all cultures can be grown in pure culture, may not be cultured at all if it’s
not caused by the microbe). Cultured microbe will cause disease when transferred to a
healthy animal (some are opportunistic pathogens and only affect those with weak immune
systems, some are not directly the cause of disease).
  What kind of evidence do scientists use to categorize and classify living things?

*** 16s rRNA, metabolic, morphology, or surface proteins

 What did the Hershey and Chase Experiment prove and disprove? Be able to
describe the method used to determine if DNA or protein was injected into a
bacteria from a bacteriophage.

The experiment proved that DNA was the genetic material of bacteriophages. Proved
bacteriophages were injecting DNA, not protein. Using phage radioactively labeled with P32
(DNA) or S35 (protein), they infected bacteria cells. Hershey and Chase wanted to know if
the bacteriophages injected DNA or protein into the bacterial cell. The bacteriophages
injected DNA. That means that P32 should be inside the cell. They blended the bacteria,
radioactive sulfur escaped the DNA pellet at the bottom and thus, the liquid was radioactive.
Blended the bacteria, radioactive phosphorus was retained in the DNA pellet at the bottom
and thus, the pellet was radioactive.

 What are the lytic and lysogenic cycles of a bacteriophage as described in class on
the board.

Feet of bacteriophage act as lock/key with external liposaccharides on bacteria surface.

Lysogenic: bacterial genome has an insert of viral DNA, just inserted, nothing happens at the
moment. This state can last forever, but can turn into lytic cycle if activated. Lytic cycle:
Viral DNA takes over machinery of bacterial cell, takes control of everything in order to
produce DNA, protein, lipase, and to break these down. Attacks bacterial genome and cuts it
randomly into smaller pieces. Bacterial genome is destroyed. Bacteria cannot do anything or
defend itself. Viral DNA used ribosomes to produce new bacteriophages (tens of thousands)
keeps producing. DNA parts self assembles into new bacteriophages. Viral DNA gets
replicated, again, and put into heads of new bacteriophages. Once they are all assembled,
they explode out and lyse the bacterial cell.

Techniques in Microbiology

 What is the difference between a simple and differential stain?

Simple stain stains all bacterial cells, allows for you to look at cell morphology. Differential
stain stains certain bacterial cells allowing you to differentiate between those and others.

 What does a Gram stain detect? How does it differentiate bacterial cells? What
color is a Gram-positive organism? What color is a Gram-negative organism?

The gram stain detects peptidoglycan/gram-positive stain: with crystal violet (appears
blue/purple). Gram-negative stain: with safranin (appears red/pink).
  What are the principle differences between a gram – and gram + bacterial cell? (this
is also covered in more detail in the external structures section)?
 What do penicillins and cephalosporin antibiotics act on with the bacterial cell? Are
gram + or gram – bacteria more susceptible to these antibiotics? Why don’t they
harm eukaryotes?

Antibiotics act on peptidoglycan. Gram + bacteria is more susceptible. Penicillins and

Cephalosporin antibiotics don’t harm eukaryotes because they do not have peptidoglycan.

 What does an acid-fast stain detect? How does it differentiate bacterial cells? What
color are acid-fast organisms after staining with an acid-fast stain? Name an acid-
fast organism.

Acid fast stain detects mycolic acid. It differentiates bacterial cells because some bacteria
have mycolic acid on the cell wall. After staining with an acid-fast stain, the acid-fast
organisms are red. Examples of acid-fast organism: Mycobacterium and Nocardia.

 Is a Schaeffer-Fulton (differential endospores green other bacterial cells red) stain a

differential stain or a simple stain? What has to be done to the proteins in the cell
membrane in order to drive the primary stain into the endospore?

The Schaeffer-Fulton stain is a differential strain. The proteins in the cell have to be heated in
order to drive the primary stain into the endospore.

 What is an endospore? Why do bacteria make endospores? What are their

characteristics? What bacterial group discussed in class is medically important and
form endospores? How does this relate to C. diff?
 What does it mean for a bacterial growth medium to be selective? Differential?
 Understand the selective and differential aspects of MacConkey's agar (MAC).
What information does MAC provide about bacteria growing on this medium?
 Understand the selective and differential aspects of Mannitol Salt agar (MSA).
What information does MSA provide about bacteria growing on this medium?
What specific bacteria can be identified in this agar (how are they clinically
important)?
 What type of clinical samples is plated on Blood agar (BAP)? What broad type of
bacteria are we looking for from a sample plated on BAP?
 What are beta, alpha and gamma hemolytic bacteria? How would they appear on
blood agar? What are some examples that were discussed in class?
 What is the difference between colony morphology and cell morphology?

Prokaryotic and Eukaryotic Cells

 Know the two basic types of cells.
 What is the 16S rRNA gene used for? What does it encode for? Why is the 16S
rRNA gene chosen as an ‘identifying characteristic’, the barcode for life?
 How do the archaea differ from eukaryotes and prokaryotes?
 What is phage typing? What characteristic of a bacteriophage make this possible?
 What is a biolog plate used for?
 Understand diffusion, osmosis, and different types of transport. What are the
different ways that molecule can move across the plasma membrane?
 What are the different transport proteins? How do they operate?
 Understand the importance of the plasma membrane in regulating the movement of
substances into and out of the cell.
 How do bacteria reproduce? How is the cytoskeleton involved in this process?
 What is the implication of bacteria not having a nucleus while eukaryotes do, in
terms of transcription and translation? Where are the ribosomes in each? What
implication does this have for the speed of transcription and translation in
prokaryotes versus eukaryotes?
 Understand osmosis and tonicity (isotonic, hypertonic, hypotonic), and be able to
solve a tonicity problem (i.e. If I describe a situation of a specific type of cell sitting
in a specific type of solution, be able to tell me which way water would move, into or
out of the cell.)
 Know the structural parts of a prokaryote and the function of those structures.
 What is the fluid mosaic model? Describe the composition of the cell wall. Know
how it is influenced by stresses.
 What are the characteristics of staphylococcus versus streptococcus?
 Understand the importance of the prokaryotic cell wall. What makes a Gram+ cell
different than a Gram- cell; how the differences impact control of microorganisms.
 What is the role of peptidoglycan? (There are several roles)
 Know the different types of glycocalyxes and the advantage that they provide to
bacteria that have them (especially in pathogenesis)
 What are the characteristics of a microbial flagella? How does the flagella move and
how do the bacteria move? Compare to cilia in eukaryotes. How is a flagella
powered?
 What is an endoflagella? What movement characteristics are induced by an
endoflagella?
 How does the shape of a bacteria possibly help in infection?
 Know the different types of surface appendages on prokaryotic cells. What does
each one do (e.g. attachment, movement, electron transport)?
 What advantage do endospores provide to bacteria?
  Know the basic shapes and arrangements of prokaryotic cells. (e.g. Staph vs. Strep),
cocci, bacilli
 What is the role of Clostridium difficile? What are the treatments?
 What is the method of action for beta-lactam antibiotics (e.g. Penicillum,
cephalosporin)
 How are endospores stained? What is a capsule stain?
 What is the role of Pili gene transfer? What do they transfer? How do Fimbriae
differ from pili?
 What is a biofilm? How do bacterial external structures aid in the development of a
biofilm? Why is it important in pathogenesis?
 What can the glycocalyx store? Why would a bacteria store this outside the cell?
How does this differ from granules?
 We have discussed competition several times. What do bacteria compete for and
what would make a bacteria ‘lose’ or ‘win’ a competition?
 Why do eukaryotes have cilia? What are some different roles of cilia?
 What is an antigen? What is an antigenic determinant? What are some examples of
antigenic determinants in bacterial external cell structures?
 What is the impact of lipid A on a human? What is techoic acid? What is the role of
Braun’s lipoprotein? What types of bacteria have them?
 Compare and contrast uniport, antiport and symport transport. Why do bacteria
transport molecules in/out of the cell? What types of molecules can diffuse passively
into the cell?
 What is the difference between active and passive transport?
 What is Nag and Nam?
 What is a murein protein, where is it found and in what type of bacteria?
 What is lipotechoic acid, who has it, where is it found?
 Where are LPS found? In what type of bacteria? What are the different portions
and what are their role? How does this affect antibiotic therapy?
 How do bacteria store nutrients?
 What is the role of fimbriae in urinary tract infections?
 What is the role of the capsule? How can a capsule ‘hide’ a cell from our immune
system? How else can it evade our immune system?
 Explain how each bacterial external structure can aid in the development of
infection.
 What is the phospholipid bilayer?
 What is the composition of the gram – lipopolysaccharide (LPS)?
 Why do jam and jellies prevent the growth of bacteria?

Microbial Metabolism
  Understand difference between catabolism / anabolism and what these reactions
have to do with metabolism.
 Understand what a redox reaction is and what this type of reaction has to do with
metabolism. Be able to identify what is being oxidized and what is being reduced.
 Understand what ATP is and its significance in metabolism.
 What are FADH2 and NADH? What is their role in metabolism?
 Understand the steps of aerobic cellular respiration to the detail that we discussed in
class (starting molecule, end product molecule, what is produced in each pathway or
subpathway. You do not need to know the name of every molecule at every substage
of these processes).
 Understand how aerobic respiration, anaerobic respiration and fermentation differ
and in what conditions each would be used.
 What are the electron acceptors in anaerobic respiration? What energetic
consequence do they have (in comparison to oxygen)? How does this influence a
bacteria’s niche?
 Be able to contrast the amount of ATP produced by the three different pathways.
 What is the electron acceptor in fermentation? What are some fermentation
products?
 What are exoenzymes? What do they do? What are the benefits but also some
potential pitfalls of bacteria producing extracellular enzymes (think competition)?
 What is the ETC? Where is it located? What is the overall function?
 Why would you take a pill of frozen feces? What is Clostridium difficile? What
causes C. diff to increase in abundance in the human gut?
 How do enzymes work, how can they be inhibited? Where are they located in
regards to the bacterial cell?
 Be able to identify the alternate electron acceptors from the redox series.
 How does protein and lipid catabolism differ from aerobic respiration? How are
they similar?

Additional material:

C. diff infections, lytic and lysogenic cycles of bacteriophage “infections” of bacteria, why
are biofilms important in disease,

These material are NOT comprehensive. Any additional material that was written solely on
the board is not necessarily included in this review.