Introduction To Microbiology

biology of the cell
Dr. Christian Rudolph

Heinz Wolff building, Room 147
christian.rudolph@brunel.ac.uk
a brief introduction
to microbiology
A very brief historic overview
Antonie van Leeuwen

hoek (16321723)
Replica of the Leeuwenhoek microscope
and a sketch of what he observed
Leeuwenhoeks observations are still of relevance

Others
Cocci
coccus
diplococcus
diplococci
encapsulated
Staphylococci
Pneumococcus
enlarged rod
Fusobacterium
streptococci
sarcina
tetrad
Vibrio
commas form
Bdellovibrio
Bacilli
coccobacillus
club rod
bacillus
diplobacilli
Corynebacteriaceae
palisades
helical form
Helicobacter pylori
corkscrews form
Borrelia burgdorferi
streptobacilli
Budding and appendaged bacteria
hypha
stalk
filamentous
spirochete

Others
Cocci
coccus
diplococcus
diplococci
encapsulated
Staphylococci
Pneumococcus
enlarged rod
Fusobacterium
streptococci
sarcina
tetrad
Vibrio
commas form
Bdellovibrio
Bacilli
coccobacillus
club rod
bacillus
diplobacilli
Corynebacteriaceae
palisades
helical form
Helicobacter pylori
corkscrews form
streptobacilli
hypha
stalk
filamentous
spirochete

Others
Cocci
coccus
diplococcus
diplococci
encapsulated
Staphylococci
Pneumococcus
enlarged rod
Fusobacterium
streptococci
sarcina
tetrad
Vibrio
commas form
Bdellovibrio
Bacilli
coccobacillus
club rod
bacillus
diplobacilli
Corynebacteriaceae
palisades
helical form
Helicobacter pylori
corkscrews form
streptobacilli
hypha
stalk
filamentous
spirochete

Others
Cocci
coccus
diplococcus
diplococci
encapsulated
Staphylococci
Pneumococcus
enlarged rod
Fusobacterium
streptococci
sarcina
tetrad
Vibrio
commas form
Bdellovibrio
Bacilli
coccobacillus
club rod
bacillus
diplobacilli
Corynebacteriaceae
palisades
helical form
Helicobacter pylori
corkscrews form
streptobacilli
hypha
stalk
filamentous
spirochete

Others
Cocci
coccus
diplococcus
diplococci
encapsulated
Staphylococci
Pneumococcus
enlarged rod
Fusobacterium
streptococci
sarcina
tetrad
Vibrio
commas form
Bdellovibrio
Bacilli
coccobacillus
club rod
bacillus
diplobacilli
Corynebacteriaceae
palisades
helical form
Helicobacter pylori
corkscrews form
streptobacilli
hypha
stalk
filamentous
spirochete
Vibrio cholerae
Staphylococcus aureus
Campylobacter jejuni
EHEC
Eukaryotic cells
Eukaryotic cells
Eukaryotic cells
Nucleus
Cell wall
Cytoplasm
Lipid bodies
Mitochondrium
Prokaryotic versus eukaryotic cells
Prokaryotic versus eukaryotic cells
Prokaryotic cells
Eukaryotic cells
No nucleus present.
The bacterial DNA forms a nucleoid.
Nucleus is formed
by a double membrane.
Prokaryotes have mostly

one single circular chromosome.
Eukaryotes have multiple chromosomes

that are linear.
Prokaryotic ribosomes are less complex

than eukaryotic ribosomes.
Eukaryotic ribosomes consist of five

rRNA species and approx. 80 different
proteins.
Prokaryotic cells do not contain

any membrane-enclosed organelles.
Eukaryotic cells contain membrane-enclosed organelles such as mitochondria,

the golgi apparatus and the endoplasmatic reticulum.
Prokaryotic cells are relatively small.
Eukaryotic cells are, as a general rule,

relatively large.
Prokaryotic cells
Prokaryotic cells
Prokaryotic cells
Cells contain enormous

amounts of DNA
Back to the 18 century

th
Where are food contaminations coming from?
open flask
broth
Groth is
observed
So with animals, some spring

from parent animals according to their kind,
whilst others grow spontaneously
and not from kindred stock; []
Aristotle, History of Animals, Book V, Part 1
The theory of spontaneous generation

was finally disproved by Louis Pasteur
Louis Pasteur
(18221895)
Heat was known to prevent bacterial growth
open flask
broth
Broth remains
sterile
Bunsen
open flask
broth
Broth remains
sterile
Bunsen
Conclusion: microbial cells

can only come from air.
open flask
broth
Broth remains
sterile
Bunsen
Conclusion: microbial cells

can only come from air.
Spontaneous generation
might require fresh air
The potential for spontaneous
generation might be destroyed
by heating
Pasteurs experiment
flask with
swan neck
Steam sterilises
inside of flask
broth
Content of flask
remains sterile
over long periods
Bunsen
Pasteurs experiment
flask with
swan neck
Steam sterilises
inside of flask
broth
Content of flask
remains sterile
over long periods
Bunsen
Pasteurs experiment
flask with
swan neck
Steam sterilises
inside of flask
broth
Content of flask
remains sterile
over long periods
Bunsen
Area contaminated
with dust and
microorganisms
Groth is
observed
in broth
Broth gets
contaminated
Pasteurs experiment
flask
Conclusion:
air alone is not
with
swan neck
sufficient to allow growth
and the potential for spon
taneous generation is not
broth
destroyed by boiling.
Steam sterilises
inside of flask
Content of flask
remains sterile
over long periods
Bunsen
Area contaminated
with dust and
microorganisms
Groth is
observed
in broth
Broth gets
contaminated
Pasteurs experiment
flask
Conclusion:
air alone is not
with
swan neck
sufficient to allow growth
and the potential for spon
taneous generation is not
broth
destroyed by boiling.
Spontaneous generation
might
require
fresh air
Steam
sterilises
inside of flask
The potential for spontaneous
generation might be destroyed
Content of flask
remains sterile
by heating
over long periods
Bunsen
Area contaminated
with dust and
microorganisms
Groth is
observed
in broth
Broth gets
contaminated
Disproving the spontaneous

generation theory allowed development
of effective sterilisation techniques
(pasteurisation of milk).
Disproving the spontaneous

generation theory allowed development
of effective sterilisation techniques
(pasteurisation of milk).
Pasteurs work was the foundation
for the germ theory and the cell theory.
The germ theory of disease
Since the 16th century it was thought that something

could be transmitted from a diseased to a healthy person.
Since the 16th century it was thought that something

could be transmitted from a diseased to a healthy person.
Diseases were called contagious,
the spreading substance was called contagion.
Robert Koch
(18431910)
Lets think about this for a minute.

Suppose I tell you that tuberculosis
is not caused by a bacterial infection.
Instead, it is caused by a mysterious
and un-identified contagion.
How do you prove this idea wrong?
Koch experiment
sick mouse
healthy mouse
microscopy of
blood sample
purification
of single colonies
injection into healthy mice
Koch experiment
sick mouse
healthy mouse
microscopy of
blood sample
purification
of single colonies
1.|
The suspected pathogenic organism

should be present in all cases of the disease
and absent from healthy animals.
Koch experiment
sick mouse
healthy mouse
microscopy of
blood sample
purification
of single colonies
2.|
The suspected pathogenic organism

should be grown to pure culture.
Just taking a sample and looking under a microscope

is rarely enough a suitable approach
Sites of the human body
that normally do not contain
any bacteria:
Spinal fluid
Blood
Joint fluid
Pleural fluid
Middle ear
Urine

any bacteria:
Spinal fluid
Blood
Joint fluid
Pleural fluid
Middle ear
Urine
Samples can get easily con

taminated. For instance, urine
is normally sterile but gets
easily contaminated upon
exiting the body.

any bacteria:
Spinal fluid
Blood
Joint fluid
Pleural fluid
Middle ear
Urine
Other bodily areas are

only healthy if they contain
a multitude of bacteria
Over 100 different bacterial
species have been found on
healthy human skin.
60% of the dry weight
of human feces are bacteria.
On average there are 2 kg of
bacteria in our intestines.
How can we separate all the bacterial

species from a sample?
Streaking bacteria to single cells on agar plates

Spread the inoculum over
a small area towards the
edge of the plate.

edge of the plate.
1. |
2. |
3. |
Sterilize the wire loop

and make several streaks.

edge of the plate.
1. |
2. |
3. |

Sterilize the loop again
and then repeat
the procedure twice.

edge of the plate.
1. |
2. |
3. |

and then repeat

edge of the plate.
1. |
2. |
3. |

and then repeat
Incubate the plates
at 37C overnight.
microscopy of
blood sample
Koch experiment
purification
of single colonies
healthy mouse
sick mouse
re-injection
into healthy mice
3.|
Cells from a pure culture

of the suspected organism
should cause disease
in a healthy animal
purification
of single colonies
Koch experiment
healthy mouse
sick mouse
re-injection
into healthy mice
4.|
The organism should be re-isolated

and shown to be the same
as the original
Koch was able to prove that germs can cause disease

(Kochs postulates).

(Kochs postulates).
He developed purification procedures for microorganisms
on solid media.

(Kochs postulates).
on solid media.
Koch was able to show that tuberculosis is caused
by the bacterium Mycobacterium tuberculosis. He was able
to purify and grow this bacterium on agar plates.

(Kochs postulates).
on solid media.
Koch was able to show that tuberculosis is caused
by the bacterium Mycobacterium tuberculosis. He was able
to purify and grow this bacterium on agar plates.
Koch developed a staining procedure
for Mycobacterium tuberculosis.
Mycobacterium tuberculosis stained by acid-fast staining procedure
Two large groups: Gram positive and negative bacteria
periplasm
peptidoglycan
pili
pili
peptidoglycan
inner
membrane
capsule
capsule
cell
membrane
outer
membrane
cytoplasm
Gram positive
periplasm
Gram negative
cytoplasm
periplasm
peptidoglycan
pili
pili
peptidoglycan
inner
membrane
capsule
capsule
cell
membrane
outer
membrane
cytoplasm
Gram positive
periplasm
Gram negative
cytoplasm
periplasm
peptidoglycan
pili
pili
peptidoglycan
inner
membrane
capsule
capsule
cell
membrane
outer
membrane
cytoplasm
Gram positive
periplasm
Gram negative
cytoplasm
periplasm
peptidoglycan
pili
pili
peptidoglycan
inner
membrane
capsule
capsule
cell
membrane
outer
membrane
cytoplasm
Gram positive
Staining technique developed

by Hans Christian Gram
(18531938)
periplasm
Gram negative
cytoplasm
periplasm
peptidoglycan
pili
pili
peptidoglycan
inner
membrane
capsule
capsule
cell
membrane
outer
membrane
cytoplasm
Gram positive
Staining technique developed

by Hans Christian Gram
(18531938)
periplasm
cytoplasm
Gram negative
Crystal violet
stains peptidoglycan
Safranin stains nuclei
Cells stained
with crystal violet
Add iodine solution,

which acts as trapping agent
Wash with ethanol
Counterstain with safranin
Cells stained
with crystal violet

Wash with ethanol
Cells stained
with crystal violet

Wash with ethanol
Cells stained
with crystal violet

Wash with ethanol
Cells stained
with crystal violet

Wash with ethanol
Mycobacterium tuberculosis has an unusual cell wall.

It is thicker than in many other bacteria, hydrophobic
and has a waxy coating.
Mycobacteria appear
Gram positive
in the Gram staining.
Mycobacterium tuberculosis has an unusual cell wall.

It is thicker than in many other bacteria, hydrophobic
and has a waxy coating.
lipoarabinomannan
Mycobacteria appear
Gram positive
in the Gram staining.
phosphatidylinositol
mannoside
outer lipids
mycolic acid
polysaccharides
peptidoglycan
plasma membrane
Ziehl-Neelsen stain (acid fast)
Stain with Carbol Fuchsin

rinse
treat for 1 min with 30% hydrochloric acid in isopropanol
counter-stain with Methylene Blue

rinse
In Mycobacterium tuberculosis
and other acid-fast bacteria
the carbol fuchsin remains in the
waxy lipids of the cell wall.

rinse
In Mycobacterium tuberculosis
and other acid-fast bacteria
the carbol fuchsin remains in the
waxy lipids of the cell wall.
Methylene blue again is a DNA

stain, so it stains all bacteria.
Kochs work was the foundation

for applied medical microbiology
and immunology.
Further ways to distinguish between bacteria in clinical isolates
selective growth media

specialised tests such as the Entertube test
selective growth media

specialised tests such as the Entertube test
immuno-tests
PCR and sequencing tests
Vibrio cholerae
Staphylococcus aureus
Campylobacter jejuni
EHEC
Bassler 2009, TED
We are harbouring
10 more bacteria
than cells
Bassler 2009, TED
AGCTTTTCAT TCTGACTGCA ACGGGCAATA TGTCTCTGTG TGGATTAAAAAAAGAGTGTC TGATAGCAGC TTCTGAACTG

GTTACCTGCC GTGAGTAAATTAAAATTTTA TTGACTTAGG TCACTAAATA CTTTAACCAA TATAGGCATAGCGCACAGAC
AGATAAAAAT TACAGAGTAC ACAACATCCA TGAAACGCATTAGCACCACC ATTACCACCA CCATCACCAT TACCACAGGT
AACGGTGCGGGCTGACGCGT ACAGGAAACA CAGAAAAAAG CCCGCACCTG ACAGTGCGGGCTTTTTTTTT CGACCAAAGG
vTAACGAGGTA ACAACCATGC GAGTGTTGAAGTTCGGCGGT ACATCAGTGG CAAATGCAGA ACGTTTTCTG CGTGTTGCCG
TATTCTGGA AAGCAATGCC AGGCAGGGGC AGGTGGCCAC CGTCCTCTCTGCCCCCGCCA AAATCACCAA CCACCTGGTG
GCGATGATTG AAAAAACCATTAGCGGCCAG GATGCTTTAC CCAATATCAG CGATGCCGAA CGTATTTTTGCCGAACTTTT
GACGGGACTC GCCGCCGCCC AGCCGGGGTT CCCGCTGGCGCAATTGAAAA CTTTCGTCGA TCAGGAATTT GCCCAAATAA
AACATGTCCTGCATGGCATT AGTTTGTTGG GGCAGTGCCC GGATAGCATC AACGCTGCGCTGATTTGCCG TGGCGAGAAA
ATGTCGATCG CCATTATGGC CGGCGTATTAGAAGCGCGCG GTCACAACGT TACTGTTATC GATCCGGTCG AAAAACTGCTG
GCAGTGGGG CATTACCTCG AATCTACCGT CGATATTGCT GAGTCCACCCGCCGTATTGC GGCAAGCCGC ATTCCGGCTG
ATCACATGGT GCTGATGGCAGGTTTCACCG CCGGTAATGA AAAAGGCGAA CTGGTGGTGC TTGGACGCAACGGTTCCGAC
TACTCTGCTG CGGTGCTGGC TGCCTGTTTA CGCGCCGATTGTTGCGAGAT TTGGACGGAC GTTGACGGGG TCTATACCTG
CGACCCGCGTCAGGTGCCCG ATGCGAGGTT GTTGAAGTCG ATGTCCTACC AGGAAGCGATGGAGCTTTCC TACTTCGGCG
CTAAAGTTCT TCACCCCCGC ACCATTACCCCCATCGCCCA GTTCCAGATC CCTTGCCTGA TTAAAAATAC CGGAAATCCTCAAGCACCAG GTACGCTCAT TGGTGCCAGC CGTGATGAAG ACGAATTACCGGTCAAGGGC ATTTCCAATC TGAATAACAT
GGCAATGTTC AGCGTTTCTGGTCCGGGGAT GAAAGGGATG GTCGGCATGG CGGCGCGCGT CTTTGCAGCGATGTCACGCG
CCCGTATTTC CGTGGTGCTG ATTACGCAAT CATCTTCCGAATACAGCATC AGTTTCTGCG TTCCACAAAG CGACTGTGTG
CGAGCTGAACGGGCAATGCA GGAAGAGTTC TACCTGGAAC TGAAAGAAGG CTTACTGGAGCCGCTGGCAG TGACGGAACG
GCTGGCCATT ATCTCGGTGG TAGGTGATGGTATGCGCACC TTGCGTGGGA TCTCGGCGAA ATTCTTTGCC GCACTGGCCCGCGCCAATAT CAACATTGTC GCCATTGCTC AGGGATCTTC TGAACGCTCAATCTCTGTCG TGGTAAATAA CGATGATGCG
ACCACTGGCG TGCGCGTTACTCATCAGATG CTGTTCAATA CCGATCAGGT TATCGAAGTG TTTGTGATTGGCGTCGGTGG
CGTTGGCGGT GCGCTGCTGG AGCAACTGAA GCGTCAGCAAAGCTGGCTGA AGAATAAACA TATCGACTTA CGTGTCTGCG
GTGTTGCCAACTCGAAGGCT CTGCTCACCA ATGTACATGG CCTTAATCTG GAAAACTGGCAGGAAGAACT GGCGCAAGCC
AAAGAGCCGT TTAATCTCGG GCGCTTAATTCGCCTCGTGA AAGAATATCA TCTGCTGAAC CCGGTCATTG TTGACTGCACTTCCAGCCAG GCAGTGGCGG ATCAATATGC CGACTTCCTG CGCGAAGGTTTCCACGTTGT CACGCCGAAC AAAAAGGC
ACACCTCGTC GATGGATTACTACCATCAGT TGCGTTATGC GGCGGAAAAA TCGCGGCGTA AATTCCTCTATGACACCAAC
GTTGGGGCTG GATTACCGGT TATTGAGAAC CTGCAAAATCTGCTCAATGC AGGTGATGAA TTGATGAAGT TCTCCGGCAT
TCTTTCTGGTTCGCTTTCTT ATATCTTCGG CAAGTTAGAC GAAGGCATGA GTTTCTCCGAGGCGACCACG CTGGCGCGGG
AAATGGGTTA TACCGAACCG GACCCGCGAGATGATCTTTC TGGTATGGAT GTGGCGCGTA AACTATTGAT TCTCGCTCGTGAAACGGGAC GTGAACTGGA GCTGGCGGAT ATTGAAATTG AACCTGTGCTGCCCGCAGAG TTTAACGCCG AGGGTGATGT
TGCCGCTTTT ATGGCGAATCTGTCACAACT CGACGATCTC TTTGCCGCGC GCGTGGCGAA GGCCCGTGATGAAGGAAAAG
TTTTGCGCTA TGTTGGCAAT ATTGATGAAG ATGGCGTCTGCCGCGTGAAG ATTGCCGAAG TGGATGGTAA TGATCCGCTG
TTCAAAGTGAAAAATGGCGA AAACGCCCTG GCCTTCTATA GCCACTATTA TCAGCCGCTGCCGTTGGTAC TGCGCGGATA
We are harbouring
10 more bacteria
than cells
Bassler 2009, TED

We are harbouring
10 more bacteria
than cells
We are harbouring
100 more bacterial
than human genes
Bassler 2009, TED

95% of bacteria
are harmless
or beneficial to us!
We are harbouring
10 more bacteria
than cells
We are harbouring
100 more bacterial
than human genes

CTAAAGTTCT TCACCCCCGC ACCATTACCCCCATCGCCCA GTTCCAGATC CCTTGCCTGA TTAAAAATAC CGGAAATCCT& Evans (2013). ATTTCCAATC TGAATAACAT
CAAGCACCAG GTACGCTCAT TGGTGCCAGC CGTGATGAAGFang
ACGAATTACCGGTCAAGGGC
GGCAATGTTC AGCGTTTCTGGTCCGGGGAT GAAAGGGATG Nature
GTCGGCATGG
CGGCGCGCGT CTTTGCAGCGATGTCACGCG
500:53839
95% of bacteria
are harmless
or beneficial to us!
We are harbouring
10 more bacteria
than cells
People whose guts contain a low diversity

of bacteria have higher levels of body fat
and inflammation than those with high gutmicrobial richness.
We are harbouring
100 more bacterial
than human genes
Applied Microbiology
Saccharomyces cerevisiae budding or bakers yeast
Microbiology as basic biological science
Provides accessible research tools for investigating

the general nature of life processes.

All cells have much in common. Bacteria, such as
Escherichia coli, are very tractable under laboratory
conditions.

All cells have much in common. Bacteria, such as
Escherichia coli, are very tractable under laboratory
conditions.
Bacteria can be very easily manipulated,
which has led to enormous advances
in our genetic and biochemical understanding
of general cellular metabolism.
Books used for this lecture:
Books for next lecture:
A classic for medical microbiology:
A classic for molecular biology:
Boye E, Lbner-Olesen A, Skarstad K (2000).

Limiting DNA replication to once and only once.
EMBO Rep 1: 479483.
Baker TA, Bell SP (1998).

Polymerases and the replisome:
machines within machines.
Cell 92: 295305.

Introduction To Microbiology

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Introduction To Microbiology

Enviado por

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biology of the cell

Dr. Christian Rudolph

A very brief historic overview

Antonie van Leeuwen

Replica of the Leeuwenhoek microscope

and a sketch of what he observed

Leeuwenhoeks observations are still of relevance

Budding and appendaged bacteria

Leeuwenhoeks observations are still of relevance

Budding and appendaged bacteria

Leeuwenhoeks observations are still of relevance

Budding and appendaged bacteria

Leeuwenhoeks observations are still of relevance

Budding and appendaged bacteria

Leeuwenhoeks observations are still of relevance

Budding and appendaged bacteria

Prokaryotic versus eukaryotic cells

Prokaryotic versus eukaryotic cells

Prokaryotes have mostly

Eukaryotes have multiple chromosomes

Prokaryotic ribosomes are less complex

Eukaryotic ribosomes consist of five

Prokaryotic cells do not contain

Eukaryotic cells contain membrane-enclosed organelles such as mitochondria,

Prokaryotic cells are relatively small.

Eukaryotic cells are, as a general rule,

Cells contain enormous

Back to the 18 century

Where are food contaminations coming from?

So with animals, some spring

Aristotle, History of Animals, Book V, Part 1

The theory of spontaneous generation

Heat was known to prevent bacterial growth

Heat was known to prevent bacterial growth

Conclusion: microbial cells

Heat was known to prevent bacterial growth

Conclusion: microbial cells

Disproving the spontaneous

Disproving the spontaneous

The germ theory of disease

Since the 16th century it was thought that something

The germ theory of disease

Since the 16th century it was thought that something

The germ theory of disease

Lets think about this for a minute.

injection into healthy mice

injection into healthy mice

The suspected pathogenic organism

injection into healthy mice

The suspected pathogenic organism

Just taking a sample and looking under a microscope

Just taking a sample and looking under a microscope

Samples can get easily con

Just taking a sample and looking under a microscope

Other bodily areas are

How can we separate all the bacterial

Streaking bacteria to single cells on agar plates

Streaking bacteria to single cells on agar plates

Sterilize the wire loop

Streaking bacteria to single cells on agar plates

Sterilize the wire loop

Streaking bacteria to single cells on agar plates

Sterilize the wire loop