Medical Microbiology

The Bacterial Structures

Key laboratory of Medical Molecular Virology
Ministry of Education and Ministry of Health
Shanghai Medical College of Fudan University

H
How
tto use th
the ttextbook,
tb k it is
i nott a easy book
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b i
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2

Key Words
Prokaryotic
Eukaryotic
Eubacteria (Bacteria)
Archaebacteria (Archaea)
Ch
Chromosome
Gram staining
Gram negative
Gram positive

Cell wall
Peptidoglycan
(murein, mucopeptide)
Outer membrane (LPS )
Cell membrane
Spheroplast/protoplast
L form
Flagella (Chemotaxis)
Pili (fimbriae)
Capsule (slime layer, glycocalyx)
Spore (resistant)

EUKARYOTES
-fungi
-plants
-animals

PROKARYOTES

BACTERIA

ARCHAEA

Chapter 3
3, p
p.41
41
Classification of Bacteria
Table 3-1 Taxonomic Rank

Eubacter-"True"
E
b t "T " bacteria
b t i
-human pathogens
-clinical
clinical or environmental
-one kingdom
Archaea
-Environmental organisms

Characteristic Prokaryotic

Eukaryotic

Drug targets?

Size (diameter) 0.1-2.0

0 1 2 0 m

10 100 m
10-100

homework

Nucleus

Nucleoid,
Nucleus
no nucleoli, no membrane

Organelles

Absent

Present

Glycocalyx

Capsule or slime layer

In some cell

Cell wall

Usually present
-peptidoglycan
peptidoglycan

Most no
-celluose
celluose /chitin

Plasma
membrane

Lack cholesterol

cholesterol

Ribosome

70S : 30S (16S rRNA)

50S (5S & 23S rRNA)

S=sedimentation
80S 40S/60S
70S(mitochondria) coefficient

Chromosome

Single circular
circular Haploid

Diploid

Cell division

Binary fission

Mitosis

S
Sexual
lR
Rec.

No, H
N
Horizontal
i
t l ttransfer
f off
DNA

M i i
Meiosis
5

Eukaryotic cell

Prokaryotic cell

Gram +
Rough
R
h endoplasmic
d l
i
reticulum
Nucleus

C ll membrane
Cell
b

Flagellum

Nucleoid

C ll wall
Cell
ll

Gram Pili

Granule

Cytoplasm
y p
Mitochondria

Capsule
Cell (inner) membrane
Ribosomes

Outer membrane

Cell wall

Size of Bacteria
Average bacteria 0.5 - 2.0 um in (- microsopes)
-- RBC is 7.5 um in diam.
Surface Area to Volume is 3:1
-- Typical Eukaryote Cell SA/Vol is 0.3:1
Nutirtion enters through SA, quickly reaches all parts of
bacteria
-- Eukaroytes need structures & organelles
Light microscope: Bright field
field microscope
100xobjective lens Phased contrast microscope
10x ocular lens
Dark-field microscope
Fluorescence microscope
Electron microscope
Bacteria are transparent
p
Scan electron microscope
Confocal microscope
Chapter 2, p. 9

Shapes of Bacteria
Bacteria are classified by shape into 3 groups:

Spiral:
Spirilla, Spirillum
Rod-shaped:
bacilli, bacillus
Round:
Cocci, coccus

Shapes
p of Bacteria
Coccus
Chain = Streptoccus
Cluster = Staphylococcus
p y
Diplopcoccus
Bacillus
Chain = Streptobacillus
Coccobacillus
Vibrio = curved
Spirillum
Spirochete

B ill
Bacillus

Spiral bacterium

Vibrio

Spirillum

Helicobaterium

Bacterial Structures
Cell Wall
-Lipopolysaccharides
Li
l
h id
-Teichoic Acids
Cytoplasm
-Inclusions
Ribosomes
Nucleoid
-Chromosome & Plasmids
Capsule
Flagella
Pili
Spores

p 2,, p.
p 21
Chapter
Cell Wall

The Cell Wall

- outside of cell
membrane
- rigid, protecting cell
from osmotic lysis
y

Hans Christian Gram

Gram staining
1884

Gram Positive

Gram Negative

Grams Serendipitous Stain,

still forms the basis for
identification of bacteria

p. 36

The Cell Wall

I am aware that as yet it is very

defective and imperfect
Hans Christian Gram

The Cell Wall

Peptidoglycan
G+ Bacteria (~90%)

G- Bacteria(~10%)

The Cell Wall

Peptidoglycan
Peptidoglycan Polymer (amino acids + sugars)
Unique to bacteria
Sugars; NAG & NAM (backbone)
N-acetylglucosamine
N-acetylmuramic acid
(The same in all bacterial species)

Tetrapeptide (vary from species)

-D form of Amino acids (not L form)
Hard to break down D form
Pentapeptide (vary from species)

- Amino acids cross link NAG & NAM

Fig. 2-15
p 23
p.23

The Cell Wall

Peptidoglycan

More than 40 sheets

in Gram positive bacteria

Only 1-2
1 2 sheets
In Gram negative bacteria

Gram+ Peptidoglycan
p
gy

17

The Cell Wall

Gram- Peptidoglycan

Fig. 2-15
p.23

G+

Peptidoglycan

G- Peptidoglycan

Peptidoglycan recognition protein, PGRP

19

The Cell Wall

E. coli
DAP

The Cell Wall

Peptidoglycan synthesis

Bacitracin

Bactoprenol phosphate transports NAM and NAG across

the cell membrane in the synthesis of peptidoglycan
Figure 6-19 p.88

GRAM POSITIVE CELL WALL

Special components: Teichoic acid TA (p.22)
Teichoic acid (WTA wall associated)
Lipoteichoic acid (LTA, membrane associated)
-Negatively charged
-Surface
S f
antigen,
ti
attachment
tt h
t off bacteria
b t i to
t animal
i l cells
ll
Lipoteichoic acid

Peptidoglycan-teichoic acid

Peptidoglycan

Cytoplasmic membrane

Figure 2-16 p.24

Cytoplasm

22

GRAM NEGATIVE CELL WALL

Special components: outer membraneLPSlipoprotein
Outer Membrane
(Major permeability barrier)

Porin

Lipopolysaccharide

B
Braun
li
lipoprotein
t i

Peptidoglycan

Inner (cytoplasmic) membrane

Periplasmic binding protein

Cytoplasm

Permease

23
Figure 2-17 p.25

Fi
Figure
2
2-17
17 p.25
25

Outer Membrane
Gram negative bacteria
major permeability barrier
space between inner and outer membrane
periplasmic
i l
i space
store degradative enzymes
Gram positive bacteria : no periplasmic space

Cell Wall Summary

Unique to bacteria
Determine shape of bacteria
(L form bacterias shape ?)
Strength prevents osmotic rupture
G+ -peptidoglycan +TA
G- -out membrane (LPS) + peptidoglycan
Some
e antibiotics
a b o cs e
effect
ec d
directly:
ec y
So
lysozyme
Penicillin

27

Wall-less forms (bacteria)

Result from action of:

y
y
lytic
y for cell wall
- Lysozyme
- antibiotics block peptidoglycan biosynthesis

In osmotically protective media (isotonic)

G- spheroplasts (with outer membrane)
- protoplasts (no outer membrane)
G+

If wall-less bacteria can g

grow and divide
L forms bacteria
chronic infection
Induced by antibiotic (penicillin)
-resistant
i t t to
t antibiotic
tibi ti ttreatment
t
t
-reversion (to normal with wall)
-relapses of the overt infection
drug resistant

29

Cell Membrane

Bilayer Phospholipid
Water can penetrate
Flexible
Not strong, ruptures easily
Osmotic
O
ti pressure created
t d by
b cytoplasm
t l

Cytoplasm
80% Water,
Water 20% Salts-Proteins)
Salts Proteins)
Osmotic Shock important

Inclusion body
-granules
granules for identification of bacteria
Chromosome
Plasmids
No organelles (Mitochondria
(Mitochondria, Golgi
Golgi, etc
etc.))

Nuclear material
Chromosome
circular Haploid
circular,
Advantages of 1N DNA over 2N DNA
-more
more efficient,
efficient grows quicker
-mutations allow adaptation to environment quicker
Plasmids
Extra-chromosomal DNA
I d
Independent
d
replication
li i
multiple copy numberhorizontal transfer
coding
- pathogenesis factors
- antibiotic resistance factors-superbug

32

Ribosome
Protein synthesis
Targets of antibiotics

Streptomycin
p
y

70S :
30S (16S rRNA)
50S ((5S & 23S rRNA))

Erythromycin

Capsules and slime layers

Envelope outside cell wall

Well defined: capsule
N td
Not
defined:
fi d slime
li
llayer or glycocalyx
l
l polysaccharide on external surface
Usually polysaccharide (Table 2-1, p30), but
often lost during in vitro culture
Protective in vivo
-adhere bacteria to surface
S mutans
S.
t
and
d enamell off tteeth
th
-prevent phagocytosis
complement cant
can t penetrate sugars
34

Fl
Flagella
ll
Some
So e bacte
bacteria
aa
are
e motile
ot e - movement
o e e t
Arrangement basis for classification
Monotrichous; 1 flagella
Lophotrichous; tuft at one end
Amphitrichous; both ends
Peritrichous; all around bacteria

Chapter 4

FLAGELLA

Locomotory
oco oto y o
organellesga e es flagella
age a
Swarming occurs with some bacteria
-Spread
Spread across Petri Dish
Proteus species most evident
Sense environment
Chemotaxis
-respond to food/poison
or unfriendly environments

Proteus

 Flagella
g

embedded in cell membrane

project as strand
Flagellin (protein) subunits
move cell by propeller like action

E. Coli with flagella

E
Shigela no flagella

38

Pili (fimbriae)
Short protein appendages (hair like)
smaller than flagella
pilins protein-vary
Adhere bacteria to host epithelium
E. coli has numerous types
K88, K99, F41, etc.
Anti-pili antibodies to block adherence
F-pilus
p
((Fertilityy factor))
-Used in conjugation (sex conjugation )
-Exchange
g of g
genetic information
Flotation, increase boyancy
Pellicle (scum on water)
More oxygen on surface

F Pilus for Conjugation

F-Pilus

Endospores (spores)
Dormant cell (non-reproductive structure )
-a thick celled structure formed inside the cell,
-encloses all the nuclear materials and some cytoplasm
Location important in classification
Central, Subterminal, Terminal
Sporulation (Fig. 2-28)
Contain calcium dipicolinate in core
Resistant to adverse conditions
Heat, irradiation, cold, organic solvents, extremely dry
g >1 hr still viable
Boiling
Sterilization, autoclave
allows the bacteria to survive for many years
There are viable
Th
i bl b
bacterial
t i l spores th
thatt h
have b
been ffound
d th
thatt are 40 million
illi
years old on Earth
-Steinn Sigurdsson

Endospores

Produced when starvedunder unfavorable condition

Bacillus anthracis form spores
p
in O2
anthrax-corpse - no necropsy
germination vegetative form (reproductive)
Dormant -germination
Activation conditions ?
(home work, -related
related with medical practice, Clostridium
tetani -tetenus )
- Bacillus stearothermophilus -spores
spores
Used for quality control of heat sterilization equipment
- Bacillus anthracis - spores (Bacillus and Clostridium)
Used in biological warfare

Bacillus anthracis G+
Aerobic
e ob c spo
spore-forming
e o
g bacte
bacteria
a
(form spores in O2 )

2011 911 attackk

Starting1week
g
after the
9/11/2001 attack,
letters containing anthrax
were sentt to
t media
di offices
ffi
and to Senators Tom Daschle
and Patrick Leahy

Summary : Bacterial Structures

Capsules and slime layers

Endospores (spores)
47

Review questions
1. In p38: question 2, 5, 6, 7, and give the explains
2 Summary the differences between the cell wall of
2.
G+bacteria and G-bacteria (related with medical practice)
3. Why penicillin and lysozame have less effect on Gbacte a
bacteria?
4. Spore germination activation conditions (related with
medical practice, Clostridium tetani -tetenus )
5. Explain which bacterial structure can be as an
antibiotics target.

49

The Cell Wall

I Gram-positive
In
G
iti b
bacteria,
t i th
the purple
l crystal
t l violet
i l t stain
t i iis ttrapped
db
by
the layer of peptidoglycan which forms the outer layer of the cell. In
Gram-negative bacteria
bacteria, the outer membrane of lipopolysaccharides
prevents the stain from reaching the peptidoglycan layer. The outer
permeabilized by
y ethanol treatment,, and the pink
p
membrane is then p
safranin counterstain is trapped by the peptidoglycan layer.

51

Peptidoglycan
synthesis and
antibiotics

Teichoic acid:
Polysaccharides of glycerol phosphate or ribitol
phosphate
p
p
linked via p
phosphodiester
p
bonds

53