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MCB 4403
PROKARYOTIC BIOLOGY
Lecture 2. Microbiology, a Historic Perspective

Life expectancy in the USA, 1900-98
Crude Mortality Rates for All Causes, Noninfectious
Causes, and Infectious Diseases
40 states have health
departments
Last human-to-human
transmission of plague
First use of
penicillin
Salk polio vaccine
introduced
First case of AIDS
First continuous use of
chlorine in municipal water
in US
1918 Spanish Flu
influenza pandemic
What Is a Microbe?

Microbes are
organisms that
are too small to
be seen with the
unaided eye.
Units of measurement
! Unit is meter (m).
! 1 !m = 10
6
m = 10
3
mm
! 1 nm = 10
9
m = 10
6
mm
! 1000 nm = 1 !m
! 0.001 !m = 1 nm
! unaided eye: resolution is
0.4 mm (400 m)
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Type of Microbial Cells
Prokaryotic cells
lack a true membrane-delimited nucleus
Eukaryotic cells
have a membrane-enclosed nucleus
are more complex morphologically
are usually larger than prokaryotic cells
! Microbes include members of the three
domains of life
Figure 1.5
Classification Schemes
Three domain system, based on
a comparison of ribosomal RNA
genes, divides microorganisms
into
Bacteria,
Archaea
Eukarya (eukaryotes)
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Carl Woese
Domain Bacteria
Usually single-celled
Majority have cell wall with peptidoglycan
Most lack a membrane-bound nucleus
Ubiquitous and some live in extreme
environments
Cyanobacteria produce significant amounts of
oxygen
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Domain Archaea
Distinguished from Bacteria by unique rRNA
(16S) gene sequences
Lack peptidoglycan in cell walls
Have unique membrane lipids
Some have unusual metabolic characteristics
Archaeal ribosomes are similar to eukaryotic
ribosomes
Many live in extreme environments (eg. high
temperature)
Domain Eukarya - Eukaryotic
Protists generally larger than Bacteria and
Archaea
algae photosynthetic
protozoa may be motile, hunters, grazers
slime molds two life cycle stages
water molds devastating disease in plants
Fungi
yeast - unicellular
mold - multicellular
Acellular Infectious Agents
Viruses
smallest of all microbes (?)
requires host cell to replicate
some cause diseases including cancers
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viruses on earth
>10
6
viral particles per ml in seawater
everyone is infected by herpesviruses.
we eat and breathe billions of viruses regularly.
Viroids and virusoids
infectious agents composed of RNA (eg. HDV)
Prions infectious proteins
Viruses
Acellular.
DNA or RNA genome.
Genome is surrounded by a
protein coat called capsid.
Coat may be enclosed in a lipid
envelope.
Viruses are replicated only when
they are in a living host cell
(intracellular parasites).
envelope

capsid
Nucleic acid
What Is a Microbe?

Microbes are living creatures (except for viruses?).
Have proteins, nucleic acids, lipid, sugars (same as us
chemically).
Metabolize, grow, reproduce, response to environment (signs of
life).
Microbial genomes (many are sequenced)
Genome = organisms total genetic content.
The phage "-X174 (5368 bps) was sequenced by Fred
Sanger in 1977.
The complete genome sequence of a cellular microbe is
Haemophilus influenza ( 2 million bps, 1,700 genes).
First draft of human genome was published in 2000.
Microbes have greatest diversity of genomes
Important for understanding evolution.
Whose genome has been sequenced first?

http/www.jcvi.org/
Have you heard these?
Metagenomics
o Metagenomics (also Environmental Genomics, Ecogenomics or
Community Genomics) is the study of genetic material recovered
directly from environmental samples.
o http://en.wikipedia.org/wiki/Metagenomics
Synthetic biology
o http://en.wikipedia.org/wiki/Synthetic_biology
Reconstruction of the 1918 influenza virus
By Jeff Taubenberger and Johan Hultin
http://www.npr.org/templates/story/story.php?storyId=4946718
Gene sequencing
Gene reconstruction
Pathological specimen
(circa 1918)
.
.
.
.
.
Reverse genetics, Synthetic Biology
Experiment in Tissue culture
And Animal models
PCR
A mosquito in amber.
1918 Flu tissue specimen
Site of a mass grave, Brevig Mission, Seward Peninsula, Alaska
Reconstruction of the 1918 influenza virus
By Jeff Taubenberger and Johan Hultin
Discovery of Microbes

Light microscope invented in 1600s
Quality improved continuously
mid-1600s: Robert Hooke observes
small eukaryotes
1676: van Leeuwenhoek discovers
bacteria
Microbes Are Living Organisms

Microbes arise only from other microbes
No spontaneous generation
1688: Redi shows that flies do not spontaneously
generate
1861: Louis Pasteur shows that microbes do not
grow in liquid until introduced from outside
No growth
Flask neck broken,
bacteria fall into and
grow in medium
Louis Pasteur

Disapproval of spontaneous
generation.
Microbes cause fermentation.
Early vaccine development.
Pasteurization.
o 63C 30 min
o 72C 15 sec (HTST).
o 140C 1 sec (UHT).
o Kill pathogen and spoilage microbes.
o #sterilization.
Louis Pasteur
Germ Theory of Disease

Observations:
Germs can infect and grow on food.
Hypothesis:
Can germs infect and grow on people?
i.e. Do germs cause disease?
Hypothesis is testable:
Are germs can be found in infected tissue?
Can transmission of germs cause disease?
Kochs Postulates

Provides means of testing hypothesis:
Does this germ cause that disease?
Organism must meet 4 criteria:
1. Microbe always present in diseased
Absent in healthy
2. Microbe is grown in pure culture
No contamination of other microbes.
3. Introduce pure microbe into healthy individual
Individual becomes sick (same disease)
4. Same microbe re-isolated from now-sick individual
Robert Koch
Figure 1.18
Limitations of Kochs Postulates
Some organisms cannot be grown in pure culture.
Using humans in completing the postulates is
unethical (no animal model).
Cofactors are needed for disease development
(ubiquitous pathogen).
Some diseases develop extremely slow.
Molecular and genetic evidence may replace and
overcome these limits.
" Beginning with Pasteurs work,
discoveries included the relationship
between microbes and disease, immunity,
and antimicrobial drugs
The Golden Age of Microbiology
Corollary to Germ Theory

Stop germ transmission, stop disease spread
Kill germ, prevent disease
Antiseptics
1865: Antiseptic surgery
Joseph Lister

Antibiotics
1929-1941: Penicillin
Alexander Fleming
Many newer antibiotics
Bacteria become resistant
Figure 1.21
Corollary to Germ Theory

Stop germ transmission, stop disease spread
Stop spread of germs
Epidemiology, public health measures

Resistant individuals prevent spread of germs
1798: Vaccination with cowpox prevents smallpox
Turkish physicians, Lady Montagu, Edward Jenner
Microbes Shape Human History

Microbial diseases change history
Black plague in Europe
Smallpox in Americas
Microbes affect food availability
Destroy crops, preserve food
bread, wine, cheese
Chocolate!
Microbial Ecology and Evolution

Most microbes dont grow on typical medium
Many live in varied conditions
Anaerobic
bottom of swamp, in our gut
High pressure
Bottom of ocean
Hot or cold temperatures
Below 0C to 113C
No organic carbon
Use light for energy, CO
2
for carbon
Microbes existed before animals, plants
Earth had no oxygen in atmosphere
Microbial Ecology

Microbes cycle most elements on earth
Nitrogen cycle
Bacteria fix N
2
to NH
4
Nitrify NH
3
to NO
3
-
Carbon cycle
Photosynthetic microbes
fix most carbon
Many other conversions
Sulfur cycle
Phosphorus cycle
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Genetics and DNA Revolution

Molecular genetics depends on bacteria
Concept of gene proposed for bacteria
DNA structure
Genetic code
Transcription, translation
Restriction enzymes
Recombinant DNA
Cloning
PCR reaction
E. coli has best understood genome
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Amazing usage of microbiology:
Botox
! Botox is a neurotoxin made by
bacterium Clostridium botulinum.
! Botulinum (Latin botulus =
"sausage) toxin is one of the
most poisonous naturally
occurring substances in the world.
! A small dose of Botox decreases
muscle activity, so the muscle is
unable to contract for a period of
4 to 6 months.
The Importance of Microorganisms
Most populous and diverse group of
organisms
Found everywhere on the planet
Play a major role in recycling essential
elements
Source of nutrients and some carry out
photosynthesis
Benefit society by their production of food,
beverages, antibiotics, and vitamins
Some cause disease in plants and animals
Summary
What are microbes? Three domains of life.
History of microbiology (germ theory, golden
age, important persons).
Medical microbiology, infectious diseases, and
human history.
Applied and environmental microbiology.
Contribution of microbiology to modern
molecular biology.
New developments in microbiology.

Key terms you should know:

Alexander Fleming, antibiotic (penicillin),
antiseptic, archaea, aseptic, autoclave, bacteria,
colonies, eukaryote, genome, Golden age of
microbiology, Joseph Lister, Kochs postulates, Louis
Pasteur, microbe, Petri dish, prion, prokaryote, pure
culture, recombinant DNA, Robert Koch, Spanish flu,
spontaneous generation, Swan-necked flask, unit of
measurement, virus.
In 1918, a pandemic of ___________ killed ~
40 million people all over the world.

A) Pneumonia
B) Influenza
C) AIDS
D) Tuberculosis
E) Cholera