Microbiology: Theory and Lab Skills

Outcome 1
Describe and explain aspects of prokaryotic micro-organisms

C Larkin after M. McIlroy/SQA/Others

The Discovery of Microorganisms

Microbiology began with the microscope
Robert Hooke (16351703): the first to describe microbes
Illustrated the fruiting structures of molds
Antoni van Leeuwenhoek (16321723): the first to describe
bacteria
Further progress required development of more powerful
microscopes
Ferdinand Cohn (18281898): founded the field of bacterial
classification and discovered bacterial endospores

Figure 1.13

Figure 1.14

Phylogenetic Tree of Life

BACTERIA

ARCHAEA

Entamoebae

Green nonsulfur
bacteria
Mitochondrion
GramProteobacteria positive
bacteria
Chloroplast
Cyanobacteria

Euryarchaeota
Methanosarcina
MethanoExtreme
Crenarchaeota bacterium
halophiles
Thermoproteus

Slime
molds

Macroorganisms

Animals

Fungi
Plants
Ciliates

Thermoplasma

Pyrodictium
Thermococcus
Nitrosopumilus

Green sulfur
bacteria

EUKARYA

Pyrolobus

Flagellates
Methanopyrus

Trichomonads

Thermotoga
Microsporidia

Thermodesulfobacterium
Diplomonads

Aquifex

C Larkin after M. McIlroy/SQA/Others

Bacteria
Bacteria are prokaryotic organisms (ie. They lack a defined
nucleus).
They can be split into two distinct groups:
1. Eubacteria true bacteria

2. Archaea lacking peptidoglycan in cell walls

C Larkin after M. McIlroy/SQA/Others

Cell wall
Cytoplasmic
membrane
Nucleoid
Cytoplasm
Plasmid

Bacteria

Ribosomes

Prokaryote

Archaea

Cell wall
Cytoplasmic
membrane
Mitochondrion
Nuclear
membrane
Nucleus
Ribosomes
Endoplasmic
reticulum
Cytoplasm
Golgi
complex
Eukarya

Eukaryote

C Larkin after M. McIlroy/SQA/Others

Bacterial Morphology (Shape)

Different bacterial species have different shapes which help
us characterise and differentiate it from other types of
micro-organisms.

C Larkin after M. McIlroy/SQA/Others

Bacterial Morphology cont.

Spheres - Cocci

Spirals

Rods - Bacilli

C Larkin after M. McIlroy/SQA/Others

Cell Division
There are two possible arrangements of bacterial cells
which do not separate after cell division.
They can arrange in two forms:
1. Clusters e.g. staphylococci
2. Chains e.g. streptococci

C Larkin after M. McIlroy/SQA/Others

Groups of Bacteria

Staphylococcus sp.
Bunch of grapes

Streptococcus sp.
Chains

C Larkin after M. McIlroy/SQA/Others

Bacterial Structure

C Larkin after M. McIlroy/SQA/Others

Bacterial Structure
Bacteria have 5 essential structural components:
1. A genome (DNA) (nucleoid region)
2. Ribosomes
3. Cell membrane
4. Cell wall
5. Surface layer (may form part of the cell wall)

C Larkin after M. McIlroy/SQA/Others

Genome
Genetic information is found on free floating
chromosomes present in the cytoplasm of the cell
in a region called the nucleoid.
Smaller pieces of DNA exist in the cell in the form
of plasmids.
This combination of chromosomes and plasmids
makes the genome of the cell.

C Larkin after M. McIlroy/SQA/Others

M.McIlroy after SQA and others (2013)

Ribosomes
Bacterial ribosomes are smaller than ribosomes
found in eukaryotic cells.
They are the site of translation during protein
synthesis ie. When the proteins are formed.
50S Ribosome Large
30S Ribosome Small

Differ from eukaryotic ribosomes

target for antibiotics eg Neomycin or
Tetracyclin
C Larkin after M. McIlroy/SQA/Others

C Larkin after M. McIlroy/SQA/Others

Bacterial Outer Layer

C Larkin after M. McIlroy/SQA/Others

Gram Staining Hans C. Gram

1853 - 1938

Bacteria can be classified depending on the content of a

complex molecule called peptidoglycan in their cell wall.
1. Gram positive bacteria contain lots of peptidoglycan
(20-30nm). They will retain the purple crystal violet dye.
2. Gram negative bacteria contain little peptidoglycan (13nm) and retain the pink counterstain safranin.
C Larkin after M. McIlroy/SQA/Others

Gram Staining Cont

C Larkin after M. McIlroy/SQA/Others

Gram Staining Cont

C Larkin after M. McIlroy/SQA/Others

C Larkin after M. McIlroy/SQA/Others

Peptidoglycan
Peptidoglycan is a large polymer that consists of a
backbone of alternating NAG and NAM molecules.
N-AcetylGlucosamine and N-AcetylMuramic acid

C Larkin after M. McIlroy/SQA/Others

N-Acetyl
group

Peptide
cross-links

Lysozymesensitive
bond

Glycan tetrapeptide

N-Acetylglucosamine ( G ) N-Acetylmuramic acid ( M )

L-Alanine
D-Glutamic acid
Diaminopimelic
acid
D-Alanine

Figure 2.25

Function of Peptidoglycan
Involved in shape and integrity.

C Larkin after M. McIlroy/SQA/Others

Gram Positive Cell Wall

Gram positive cell walls also contain teichoic acid that
confers a negative charge.
The main role of teichoic acid is to:
Provide rigidity to the cell wall

Play a role in the attachment of bacterial cells to tissue

surfaces
Some may act as receptors for viruses that infect
bacteria

C Larkin after M. McIlroy/SQA/Others

Function of Teichoic Acids

Confer negative charge and antigenicity (ability to
bind to surface receptors) in Gram positive
bacterial cells.
Aid attachment of bacterial cells to tissues.

C Larkin after M. McIlroy/SQA/Others

Gram Negative Cell Walls

Gram
negative
cell
walls
also
contain
lipopolysaccahride (LPS) which is an endotoxin.
Endotoxins play a role in infection by Gram
negative bacteria.
Lipid A is the toxic component of the LPS
endotoxin.

C Larkin after M. McIlroy/SQA/Others

Structure of Lipopolysaccharide

C Larkin after M. McIlroy/SQA/Others

Function of LPS (Endotoxin)

Confers toxicity (Lipid A).
Determines virulence (organisms ability to cause
disease).

C Larkin after M. McIlroy/SQA/Others

Cell Wall Function

1. Gives bacterial cell shape.
2. Provides protection from osmotic lysis and toxic
substances.
3. Plays a role in cell division and motility.
4. Pathogenic bacteria have components which
contribute to pathogenicity.
5. Site of action of many antibiotics.
C Larkin after M. McIlroy/SQA/Others

Function of Outer Membrane

1. Forms the outer limit of the periplasmic space.
2. Helps the bacterial cell evade phagocytosis and
the action of complement (activation of the
immune response)
3. Provides a permeability barrier
4. Allows passage of nutrients through porins.

5. Receptor for F pilus (conjugation)

6. Attachment site for bacterial viruses
C Larkin after M. McIlroy/SQA/Others

Function of Cell Membrane

1. Selectively permeable membrane
2. Separates the cytoplasm and the unit bacterial
cell from its environment.
3. Provides transport systems for movement of
large or charged molecules across the
membrane.

C Larkin after M. McIlroy/SQA/Others

Function of the Cell Membrane Cont

4. Site of respiration and energy generation.
5. In photosynthetic bacteria it is the site of
photosynthesis.
6. Site of biosynthesis of lipids and cell wall
components eg peptidoglycan.
7. Co-ordination of DNA replication.
8. Contains receptor molecules which allow the
bacterial cell to respond to chemicals in the
environment ie chemotaxis.
C Larkin after M. McIlroy/SQA/Others

Bacterial Cell Parts Continued

In addition to the cell components discussed
bacteria also have:
Mesosomes
Inclusion bodies
Periplasmic Space

Flagella
Cillia
C Larkin after M. McIlroy/SQA/Others

Function of Mesosomes
Mesosomes are folds in the bacterial cell
membrane found in gram positive and gram
negative bacteria.
They have various functions including:

involved in DNA replication and segregation

involved in cell wall synthesis
play a role in respiration and photosynthesis
C Larkin after M. McIlroy/SQA/Others

Inclusion Bodies
Inclusion bodies are bodies of reserve material within the
cell.
They have different roles depending on the type of cell they
are in. E.g. gas vesicles provide buoyancy in water.

C Larkin after M. McIlroy/SQA/Others

Periplasmic Space
Periplasmic space is the border between the cell membrane
and the cell wall.
Gram positive bacteria; either do not have a periplasmic
space or the periplasmic space is small.
Gram negative bacteria; have large periplasmic spaces.

C Larkin after M. McIlroy/SQA/Others

Comparison of Gram (+) and Gram (-) outer

membranes

C Larkin after M. McIlroy/SQA/Others

Flagella
Flagella are filamentous structures made of the protein
flagellin.
They allow the cell to be motile and can extend beyond the
cell wall up to 10 times the length of the cell.
The flagella rotates at up to 60 revolutions per second.

C Larkin after M. McIlroy/SQA/Others

Flagella

Fimbriae

Figure 2.33

Bacterial motion

http://www.youtube.com/watch?v=4hexn-DtSt4
C Larkin after M. McIlroy/SQA/Others

Flagellar Arrangement
Flagellum distribution can be one of four types:
1. Monotrichous one flagellum located at one pole of the
cell.
2. Amphitrichous a single flagellum at both poles of the
cell.
3. Lophotrichous a short cluster of flagella at one or both
poles of the cell.
4. Peritrichous lateral flagella distributed over the whole
surface of the cell.
C Larkin after M. McIlroy/SQA/Others

Flagellar Arrangement Cont...

The flagellar arrangement
allows us to differentiate
different types of bacteria.
Eg. Pseudomonas spp.
Have polar flagellar
Samonella spp. Have
peritrichous flagellar

C Larkin after M. McIlroy/SQA/Others

Pili
Pili or fimbriae are short hair like structures found in both
gram negative and gram positive bacteria.
They have 2 distinct functions.
1. Common pili
(attachment)

involved

in

specific

adherence

1. F pili - bring about genetic exchange or mating in

bacteria through a process called conjugation.

C Larkin after M. McIlroy/SQA/Others

Biofilms
A biofilm is formed when bacteria attach to a surface,
produce slime and then divide forming microcolonies within
the slime layer.
The biofilm provides an enriched, protected environment for
themselves and other bacteria forming a complex bacterial
environment.

http://www.youtube.com/watch?v=OdzJESVgz10

C Larkin after M. McIlroy/SQA/Others

Function of Biofilms
1. Aid in attachment of bacterial cells to surfaces (including
cells).
2. Protect bacterial cells from engulfment by protozoa and
by white blood cells (phagocytosis).
3. Protect against attack by antimicrobial agents.
4. Exclude viruses and other toxic material eg detergents
5. Protect bacterial cells against desiccation.
6. Aid in pathogenicity by helping bacterial cells to resist
phagocytosis by phagocytes.
C Larkin after M. McIlroy/SQA/Others

Bacterial Endospores
Bacterial Endospores are a protective, highly resistant
structure that allow bacteria to survive in hostile
environments.
They are typically gram positive bacteria with the most
notable being the Bacillus genus, and the Clostridium
genus.
Endospore are known as cryptobiotic in that they show no
signs of life, however they ensure the cell remains viable
when environmental conditions return to normal.

C Larkin after M. McIlroy/SQA/Others

C Larkin after M. McIlroy/SQA/Others

Formation of Spores

http://www.youtube.com/watch?v=NAcowliknPs
C Larkin after M. McIlroy/SQA/Others

Bacterial Endospores cont...

Bacterial endospores can survive:
high temperature eg boiling in water at 100C for 2 hours
or more
irradiation

desiccation (eg 70% alcohol for 20 years)

strong acids
organic solvents
disinfectants
C Larkin after M. McIlroy/SQA/Others

Bacteria Reproduction
Bacteria reproduce asexually by binary fission, results in
two identical daughter cells.

C Larkin after M.McIlroy/SQA/Others, 2016/17

C Larkin after M. McIlroy/SQA/Others

Binary Fission
1.

The chromosome duplicates

2. The cell grows and elongates new cell wall and
membrane is produced
3. The plasma membrane pinches inward at the centre of
the cell, when the nuclear material has been evenly
distributed, the cell wall thickens and grows inward to
separate the dividing cell into 2 new daughter cells. (No
spindles or asters are present as in eukaryotes)
4. The daughter cells are genetically identical to the parent
cell.

C Larkin after M. McIlroy/SQA/Others

Questions?

C Larkin after M. McIlroy/SQA/Others