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Lecture 1

Basics of Microbiology:
Introduction to Structure and
Function of Bacteria

Dr Charlene Kahler
Rm 2.03A
L Block QE II Medical Center
email: charlene.kahler@uwa.edu.au
What can go wrong?
Toddler Tarliah O'Connell's tooth agony 'ignored' for days
Gemma Jones

From: The Daily Telegraph

February 20, 2010 1:43AM

Doctors gave her only Panadol

She lost her teeth after four days of pain

Hospital ended up giving her morphine

A TODDLER suffering a mouth infection so severe it later required surgery and teeth removal
was given only Panadol while the infection worsened at a hospital.

Tarliah O'Connell, from Penrith, NSW, was moaning in pain after four days at Nepean Hospital and the painful
infection then spread across her face.

After being given only Panadol and Nurofen, a desperately sick Tarliah was transferred to The Children's
Hospital, Westmead, last Thursday.

Her mother Raquel said Tarliah was immediately given morphine to control her terrible pain. She said her
daughter has been left so traumatised she was now scared of medical staff.

Tarliah has undergone serious facial surgery to treat the infection which even spread to her throat.
Objectives for this lecture:
At the end of this lecture, you should be able to:
Explain the history of microbiology in dental hygiene
Describe the difference between eukaryotes and bacteria
Explain the diversity of microorganisms in relation to size, shape
and arrangement.
Draw a figure showing the basic features of a bacterial cell
Describe the function of the following components of a bacterial
cell: nucleiod, cytoplasm, cytoplasmic membrane, cell wall.
Explain binary fission using a diagram
Describe the principle behind the Gram stain which
differentiates bacteria into Gram positive and Gram negative
categories.
List the structural differences and similarities of Gram negative
and positive bacterial cell walls and explain how these features
impart different properties to the cell.
What are: capsules, pili, flagella and endospores?
Microbiology
Definition: The study of microorganisms
includes bacteria, archaea, yeasts, fungi
(molds), viruses, algae, and protozoans
involves understanding
How do they spread from location to another?
Why do some cause disease and others are
beneficial?
What is their role in the environment?
How do they operate?
Microbiology and dental hygiene
1676: Anton van Leeuwenhoek
first person to observe
microorganisms (70 -300 fold mag).

QUOTE:
There are more animals living in the
scum of a mans mouth, than there are
men in a whole kingdom? Especially in
those who dont ever clean their teeth,
whereby such a stench comes from the
mouth of many of them that you can scarce
bear to talk with them.

A.V. L. Letter 39
17 September 1683
Published in Proc. Royal. Soc. XIV, No 159, p 568 1684
Important events
1498: Chinese encyclopaedia
The modern toothbrush

1819: Dr. Levi Spear Parmly


Introduced the concept of preventive oral
health practices by flossing teeth

1839: Baltimore College of Dental


Surgery
Enamel
First dental school in the world
Dental
1857: Louis Pasteur Caries
proposed the "germ theory" of disease

1889: Dr Willoughby Dayton Miller


Root
established the bacterio-chemical theory of
dental caries
http://odonto-
red.com/cariesdental.htm
Diversity of Microorganisms
Diversity of microorganisms
Prokaryotes
Bacteria*
Archaea*
Eukaryotes
Yeasts*
Fungi (molds)*
Algae
Protozoans*
Viruses*

* Causative agents of the


majority of diseases in the oral
cavity

Showing Domians and Phylums


Bacteria and Eukaryotes
Bacteria Eukaryotes (fungi, molds)
DNA not enclosed in a DNA found in a nucleus,
membrane, circular organized as linear
No organelles chromosomes
Most have cell walls Organelles are present
containing muramic acid Cell walls with no muramic
(peptidoglycan) acid
Diversity of Microorganisms: Size

The basic unit of length is the meter (m),


and all other units are fractions of a meter.
Nanometer (nm) = 10-9 = .000000001 m
Micrometer (mm) = 10-6 = .000001 m
Millimeter (mm) = 10-3 meter = .001 m
Diversity of Microorganisms: Shape
Five basic shapes
Spherical
Coccoid

Cylindrical
Rod/Bacillus
Pleiomorphic:
Spiral Actinomyces israeli
Spirillium A mixture of microbes
Helical (different colours)
(Scanning electron
Spirochete microscope, falsely
Square coloured)

Pleomorphic
Diversity of Microorganisms:
Arrangement
Determined by the plane
of cell division (binary
fission)
Rods
1 plane to produce a
single rod or chain of rods
Cocci
1 plane to produce single
cocci or diplococci
2 plane to produce a
tetrad
3 random planes to
produce a bunch
Bacteria: Binary Fission
When one bacterial cell
undergoes repeated
rounds of division on a
solid surface, it results in a
single colony composed of
identical cells
This is a very fast process
Escherichia coli
undergoes one round of
cell division in 20 min
Bacteria: Composition
WATER 70%
DRY WEIGHT 30%
Composed of:
DNA 3% (MW = 2 x 109)
RNA 12%
Protein 70%
found in:
4 6
Ribosomes (10 ) (RNA-protein particles, MW = 3 X 10 )

Enzymes

Cell Wall and membranes


Polysaccharide 5%
Lipid 6%
Phospholipid 4%
Bacteria: Structure
Nucleoid
Genetic information
Cytoplasm
Cytoplasm
Metabolism Nucleoid
Cell Wall
Ribosomes performing protein Cell membrane
synthesis
Plasma membrane
Consists of phospholipids and
proteins
Permeable
Controls exchange of gases,
nutrients etc
Cell wall (except Mycoplasma)
Rigid
Optional
Vacuoles, capsules, slime
layers, pili, flagella, endospore
Bacteria: Genetic material
Nucleoid
Double stranded Deoxyribonucleic
acid (dsDNA)
Usually Circular in single copy Relaxed
Complexed to basic proteins that
enable supercoiling
Size <0.2uM in diameter
Encodes genes which are
essential for life

Plasmids
Circular (single or multiple copies)
supercoiled dsDNA (2-200kb) Supercoiled
Replicate independently of the Unit lengths
1000 base pair (bp) units = kilobase (kb)
host chromosome
1000 kb = 1 megabase (Mb)
Carry genes which are not Escherichia coli genome = 4.6Mb or 1,400mM
essential to the host but may be
useful in certain environments
Bacteria: Cytoplasm
Cytoplasmic matrix
Located between the plasma
membrane and the nucleoid
Composition
70% water
Contains ribosomes
Composed of
proteins/ribonucleic acid
(RNA)
Two protein subunits: 30S and
50S
Essential for protein synthesis
Highly organised cytoskeleton-
like system of proteins for cell
division
Optional: a variety of inclusion
bodies (specific to different
bacteria)
Bacteria:
Cytoplasmic membrane
Composition
Amphipathic phospholipids
organised as a bilayer
Proteins
Function
Selectively permeable
membrane that determines
transport of molecules into
and out of the cell
Nutrient transport
Energy metabolism
First step in transport of
proteins out of the cell
Bacteria: Cell Wall
Function
Maintains shape
Provides protection from
Osmotic shock
Action of anti-bacterial
agents (eg. Antibiotics,
antiseptics etc)
Contain components
contributing to pathogenicity,
in particular, inflammation
Present in all bacteria
(except Mycoplasma) and
Archaea
Procedure for the Gram strain
Characteristics of Gram positive and
negative cells
Bacteria:
Components external to the Cell Wall
Variety
Capsules and Slime Layers
Pili / Fimbriae
Flagella
Roles
Protection from the environment
Adhesion
Motility
Capsules and Slime Layers
Capsule
Organized matrix firmly
anchored to the cell
Streptococcus
Slime layer mutans
Diffuse, disorganized matrix,
easily removed
Capsule
Composition
Polysaccharides
Poly amino acid
Capsule stain of
Role Streptococcus
Resist phagocytosis lactis
Protection from desiccation
Adherence
Gliding motility (slime)
Pili / Fimbriae
Short, fine, hair-like
appendages
Hollow brittle tube
Consists of multiple pilin
subunits organized as a helix
Types I-IV defined by
Length
Diameter of the fibre
Location on the cell
Polar
Peritrichous
Single, multiple
Twitching

Function motility
Adherence
Twitching motility
Gliding motility
Myxococcus
Synechocyctis
Flagella and Motility
Hair-like appendage
10-20um long
0.02 um thick
Consists of
Hollow filament composed of
flagellin protein
Basal motor structure drives rotation
of flagellum which acts like a
propeller
Different attachment locations on
cell
Polar
Peritrichous
Mono- or multiple
Role
Swimming in aqueous environments
Seeking nutrients: chemotaxis
Alternate cell structures:
Endospores
Distribution
Produced by small number of
Terminal eg
bacteria Clostridium tetani
Bacillus, Clostridium
Structure

Subterminal eg
Bacillus subtilis

Central eg
Bacillus anthracis
Endospores: Characteristics
Property Vegetative cells Endospores
Characteristics
Resist extreme conditions Typical Gram- Thick spore coat,
positive murein cortex, and
which would kill the vegetative Surface coats
cell wall peptidoglycan
cell, for example polymer core wall
Nutrient starvation
High temperatures (100oC for Microscopic
Nonrefractile Refractile
1 hr) appearance
Salt Calcium dipicolinic
Absent Present in core
Core contains dipicolinic acid acid
(15% dry weight) Cytoplasmic water
High Very low
Sporulation occurs in response activity
to limited carbon, nitrogen,
Enzymatic activity Present Absent
phosphorous
Role Macromolecular
Present Absent
synthesis
Endospore germinates in
favourable conditions to Heat resistance Low High
become a vegetative cell Resistance to
chemicals and Low High
acids
Remove them from the
Radiation resistance Low High
environment by autoclaving
Sensitivity to
Sensitive Resistant
lysozyme
Sensitivity to dyes
Sensitive Resistant
and staining
Endospores: Sporulation
Alternate Cell structures:
Conidiospores
Distribution
Produced by
Streptomyces
Response to nutrient starvation
Structure
Thin wall spores at the end of
filaments
Characteristics
Resistant to desiccation but not
heat

(Scanning electron micrograph,


Mark Buttner, Kim Findlay, John
Innes Centre
Background text
Examinable content
Microbiology. Eds: Prescott, Harley and
Klein. 6th Ed. Chapter 1 and 3.

Prescotts Microbiology. Eds.


Willey/Sherwood/Woolverton. 8th Ed.
Chapter 2- pgs 36-38, Chapter 3- pg 46-
63.
Revision questions (these are examples- so think of
the variants that can come from each one!)
Who was the first person to observe microorganisms? (another variant is who proposed the germ theory of
disease etc)
What process did Dr Levi Spear Parmly introduce to the practise of dentistry? (another variant is what was
Lois Pasteur known for?)
How many domains of life are there? What are the domains of life called?
To which domain do bacteria belong? (same can be asked for archaea, yeasts, fungi, algae, protozoans,
viruses)
Organisms in which phylum are not found in the oral cavity?
What are the key structural features of a bacterial cell?
What are the key structural features of eukayote fungi and molds/
How many planes of cell division are there in cocci? How does this affect the arrangement of bacterial
cells?
What are the steps in binary fission/
What is the major component of protein found in bacterial cells? What is the most abundant nucleic acid in
bacterial cells?
What are the four main structures of a bacterial cell? What are their roles?
What does the nucleoid/cytoplasm/cytoplasmic membrane/cell wall consist of?
How is the genetic material arranged in the bacterial nucleoid?
What are plasmids? Are plasmids in the nucleoid?
What is the structure of a phospholipid? What do phospholipids do in an aqueous environment?
What is the structure of a Gram positive cell wall?
What is the structure of a gram negative cell wall?
What are the steps in a Gram stain? What does each step do?
What is a capsule layer? How does this differ from a slime layer? What is the role of capsules and slime
layers?
Revision questions (these are examples- so think of
the variants that can come from each one!)
What is a pili/fimbriae? What is the structure of pili? What is the role of pili? How are pili located on the
bacterial cell surface?
What is a flagella? What is the structure of flagella? What is the role of flagella? How are flagella located
on the bacterial cell surface?
What is an endospore? What bacterial species produce endospores? What is the structure of an
endospore? What are the characteristics of an endospore? What triggers sporulation? What triggers
germination? How do you kill endospores?
What is a conidiospore? What bacterial species produces conidiospores? What triggers the production of
conidiospores? What is the structure of a conidiospore? How do you kill a conidiospore?