General bacteriology

Introduction

 Bacteriology: the discipline of biology related to the study of

bacteria.
• Unicellular organism

• Bacteria are prokaryotes

•Fact: Each square centimeter of your skin averages about

100,000 bacteria. A single teaspoon of topsoil contains more
than a billion (1,000,000,000) bacteria.
Bacteria Characteristics
• Some bacteria are aerobic, meaning that they require
oxygen in order to survive. Other bacteria are
anaerobes, meaning that they do not require oxygen to
survive.
• Most bacteria are harmless and offer beneficial
functions to living things and humanity.
• Bacteria can be moved by air and water currents, and
on any surface such as clothing, hands, or any object.
Bacteria contain:
• a singular, circular
piece of DNA
• tiny circular pieces
of DNA called
plasmids
• ribosomes
Bacteria
reproduce:

Asexually
(mitosis)
using binary
fission.
Bacteria reproduce:
sexually using
conjugation.
Bacteria exchange
plasmid DNA.

This is how bacteria

become antibiotic
resistant.
Bacteria have cell walls made of:
• peptidoglycan (polysaccharide linked
with chains of amino acids).
• this may be covered with an outer
membrane of lipopolysaccharide
(chain of sugar with a lipid attached).
Some bacteria:
• have a gelatinous layer called a
capsule surrounding the cell wall.
• form thick-walled endospores around
chromosomes when they are
exposed to harsh conditions
(drought, high temperatures) - these
types cause botulism
Some bacteria have:
• flagella for
locomotion.
• pili (short, thicker
outgrowths that
help cell to
attach to
surfaces)
¨Three bacterial shapes:
1. Rod shaped are called bacillus(i)
2. Sphere shaped are called coccus(i)
3. Spiral shaped are called
spirillum(I)
Gram stains:
• Important in medicine because
provides information for treatment of
bacterial disease.
•Bacteria stain either gram positive
(purple) or gram negative (pink).
•Gram positive tend to respond to
penicillin and like antibiotics.
•Gram negative respond to types of
antibiotics unrelated to penicillin.
 Gram Stain Process:

Crystal violet dye Iodine

Purple Decolorizes
Alcohol Safranin
Pink Counterstain
A Gram Stain is
usually performed
on a smear
preparation that
has been heat
fixed. One
function of
fixation is to
secure (fix) the
cells to the slide.
Gram positive = purple because of large
amount of peptidoglycan in cell wall.

Polymer consisting of sugars and amino acids that forms a mesh-

like layer outside the plasma membrane of eubacteria.
Gram negative = pink because¨
lipopolysaccharide membrane is removed by
alcohol and thin peptidoglycan layer allows dye to
escape.

(LPS) is a large molecule consisting of a lipid and a

polysaccharide (carbohydrate) joined by a covalent
bond.
• work by preventing cell wall
formation, breaking up cell
membranes, or disrupting chemical
processes.

•Antibiotics cannot treat viral

infections.
There are three types of
bacteria based on how they
obtain energy: heterotrophs,
photosynthetic, and
chemoautotrophs.
This is the
anthrax
bacterium.
1. Most are heterotrophs:
a.Decomposers – feed on and recycle organic material
• Bacteria are decomposers, they break down the chemical elements inside
other living or dead organisms. Some bacteria live in the intestines of
humans and animals
b.Pathogens – parasitic, disease-causing bacteria
• Either attack cells or secrete toxins
c.Nitrogen-fixing bacteria (Rhizobium)
• Found in nodules of soybeans, peanuts, alfalfa, and clover
• Convert atmospheric nitrogen (N2) into ammonia, called nitrogen fixation.
• Used in crop rotation
• Rhizobia bacteria convert nitrogen gas into other substances that help plants
grow.
2. Some, like Cyanobacteria, are
photosynthetic.
• These are autotrophs that use the
sun’s energy to make food.

This bacteria has

chlorophyll and uses the
process of photosynthesis
to produce food very much
like a plant.
3. Chemoautotrophs
•Obtain energy by removing electrons from
inorganic molecules such as ammonia
and methane to make food.
•Examples: Nitrobacter and Nitrosomonas
- live in soil
- have a crucial role in nitrification (turn
ammonia into nitrates, the
form of nitrogen commonly
used by plants).
Some Good Bacteria

• Lactic acid bacteria have been used to ferment or

culture foods for at least 4000 years.
• Examples: products like yogurt and cheese

• Wastewater bacteria feed on everything from

solid human waste matter to last night's leftovers.
As the bacteria eat, they convert organic matter to
carbon dioxide, releasing electrons, the basic
element of electrical current.
Taxonomy:
Naming, Classifying, and
Identifying Microorganisms
• Microbial nomenclature- naming
microorganisms
• Taxonomy- classifying living things
originated over 250 years ago with the work of
Carl von Linné
• Identification- discovering, comparing
and recording the traits of organisms so
they can be named and classified
• Levels of Classification
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 Nomenclature

• Binomial (scientific) nomenclature

• Genus – Bacillus, always capitalized

• species - subtilis, lowercase

• Both italicized or underlined

– Bacillus subtilis (B. subtilis)
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 Identification

• The process of discovering, comparing

and recording the phenetic (physical,
biochemical) and genetic traits of
organisms, thereby, placing them in a
taxonomic scheme.

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 Bacterial classification and nomenclature

• There is no official classification of bacteria, but

classification is based on several methods.
 Bacteria are classified in to different categories
in Bergey’s manual of determinative
bacteriology, (1974), and the classification is
based on
1. Morphology 8. Amino acid sequencing of proteins
2. Staining 9. Genetic composition
3. Motility
4. Growth
5. Nutritional requirement
6. Bio chemical and metabolic activity
7. Pathogenicity
 Morphology: -
 Bacteria vary widely in size, ranging from 0.2 um to
10um long
-There are three basic morphological forms/shapes
1. Spherical or coccoid/cocci- (singular –coccus)
2. Rods or bacilli (singular - bacillus)
3. Spirals or spirilla
Fig . Different bacterial morphologies A. Cocci, B. Bacilli, C.
Spiral shape
Fig. different bacterial arrangements
 Basic features of Bacterial Cell

• General property
– Typical prokaryotic cell
– Contain both DNA and RNA
– Most grow in artificial media
– Replication is by binary fission
– Contain rigid cell wall
Bacterial cell structure and Function
 Structures in bacterial cells

Structures common to Structures found in Structures found in some

most bacterial cells bacterial cells

all bacterial cells
• Flagella
• Cell membrane • Cell wall
• Pili
• Surface coating or
• Cytoplasm • Fimbriae
glycocalyx
• Ribosomes • Capsules

• One (or a few) • Slime layers

• Inclusions
chromosomes
• Actin cytoskeleton
• Endospores
 Bacterial Structure

Bacterial structure is considered at three levels.

1. Cellular element external to the cell envelope (appendages):

Flagellum, Pilus

2. Cell envelope proper:

Glycocalyx, cell wall and cell membrane

3. Cellular element enclosed with in the cell envelope:

Mesosomes, ribosome, bacterial chromosome, cytoskeleton

and cytoplasmic granules.
Fig: Bacterial cell structure
1. Cellular Element External to the Cell Envelope

A. Flagellum
• organ of locomotion/ motility containing
filaments
• Attached to cell membrane
• It is composed of protein named as flagellin
• Antigenic: the flagellar antigen is named as H
(Hauch) antigen.
Each flagellum has 3 distinct parts:
a) Filament or shaft
– It is external to the cell surface twisted helically
and connected to the hook at the cell surface.
b) Hook – connect the basal body with the
filament
c) Basal body – embedded in cell envelope and consists of a small,
central rod surrounded by a series of ring

Fig. Bacterial flagellar structure

 Flagellum … Cont’d
Flagellar arrangements
A. Atrichous: Bacteria with no
flagellum. E.g. All cocci bacteria
B. Monotrichous: Bacteria with single
polar flagellum
E.g. V. cholarae
C. Amphitrichous: Bacteria with
flagella at both poles.
Eg. Alcaligenes faecales
D. Lophotrichous: Bacteria with bunch
of flagella at one pole.
E.g. helicobacter
E. Peritrichous: Bacteria with flagella
all over their surface:.
E.g. S. typhi , E. coli
B. Pili (Fimbriae): used interchangeably

• short, hair-like structures (finer

filaments) on the surfaces of
prokaryotic cells.
• are extruding from the
cytoplasmic membrane
• are shorter and stiffer than
flagella, and slightly smaller in
diameter.
• composed of protein called pilin
Two functional types of pili may occur
independently or together on some cell.

I. Common pili
• almost always called fimbriae
• Involved in adherence and
colonization of other cells
• Inhibit phagocytosis

E.g. Streptococcus pyogenes

II. F pili or Sex pili
• Found only in Gram negative cells
• specifically involved in
bacterial conjugation, i.e
transfer of genetic
material (DNA) from
one bacterium to
another.
• E.g. E. coli
 2. Cell envelope proper
A. slime layer and capsule

• Slime layer: a less discrete

polysaccharide or matrix
which embeds the cells

• Capsule: discrete detectable

layer of polysaccharides and
proteins deposited outside
the cell wall
 Function of Capsules and slime layers

Attachment:
• These structures are thought to help cells attach to
their target environment.
 E.g: S. mutans produces a slime layer in the
presence of sucrose, which result in dental plaque.

Resistance to drying:
• inhibit water from escaping into the environment.

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Protection from phagocytic engulfment:
• The capsule protect the bacteria by making it
difficult for the phagocyte to engulf the
microbe.
 E.g. S. pneumoniae, when encapsulated is
able to kill 90% of infected individual,
when non-encapsulated not kill.

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B. Cell wall
– a non-living secretion of the cell membrane,
– Multi layered structure and constitutes about 20% of the
bacterial dry weight
– Average thickness is 0.15-0.5 m
– Fully permeable to ions, amino acids, and sugars
(Gram positive cell wall may regulate movement of
cations)
Chemical composition of the cell wall
• The rigidity of the cell wall is due to the
presence of a unique substance called
peptidoglycan layer (murein). Some times
called back bone of bacteria.
• PG is found only in bacteria and not found in
other microorganisms.
• The beta lactam drugs prevent the action of
these enzymes and inhibit bacterial growth
 Function of the cell wall
• Provides shape to the bacterium
• Gives rigidity to the organism -prevent lysis of
cells due to osmotic pressures
• Maintains cell integrity
• Protects from destructive environmental factors
• Contains receptor sites for phages
 Types of cell wall
Depending on Gram stain two generally
different groups of bacteria are demonstrated:
I. Gram positive cell wall of bacteria
• has two layers (peptidoglycan) cross linked with
teichoic acid(These polymers of glycerol phosphate
or ribitol phosphate )
• Teichoic acids the major surface antigens of the
gram-positive cell wall.
• The PG layers is much thicker than Gram negative
bacteria and is 15 – 50μm thick
• The large amount of peptidoglycan make G+ve
bacteria susceptible to the lysozyme and penicillin.
Fig. Gram-Positive
the cell wall of
bacteria
II. Gram negative cell wall of bacteria

• Is a complex than Gram positive bacterial cell wall

 Has high lipid content (lipopolysaccharide) in the
outer membrane
• Has thin peptidoglycan layer (3 – 8μm)
• Has periplasmic space
Fig. Gram-negative cell
wall of bacteria
 Comparisons
Gram +ve vs. G-ve cell walls

Fig. Gram-Positive and Gram negative cell wall

of bacteria
 Gram Stain
Differential stain that distinguishes cells
with a Gram-positive cell wall from those
with a Gram-negative cell wall
◦ Gram-positive - retain crystal violet and stain
purple
◦ Gram-negative - lose crystal violet and stain red
from safranin counterstain
Practical aid in diagnosing infection, and
guiding drug treatment

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54
 Prokaryotes - Cell Wall
From the peptidoglycan inwards all bacteria are very similar. Going further out, the
bacterial world divides into two major classes (plus a couple of odd types). These are:

Gram-positive Gram-negative
 Atypical Cell Walls
• Some bacterial groups lack typical cell wall
structure i.e. Mycobacterium and Nocardia
– Gram-positive cell wall structure with lipid
mycolic acid
• Pathogenicity and high degree of resistance to
certain chemicals and dyes
• Basis for acid-fast stain used for diagnosis of
infections caused by these microorganisms

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 Bacterial lacking/ deficient cell wall

1. Mycoplasma spp: a genus of naturally

occurring stable bacteria that lack rigid cell wall.
e.g: Mycoplasma pneumoniae and Ureaplasma
urealyticum
2. L-forms of Bacteria: These are mutant bacteria,
produced in the laboratory and body treated with
penicillin. Able to grow as:
 Protoplasts: unstable bacteria derived from Gram-
positive bacteria and totally lacking cell walls;
produced artificially by lysozyme which destroy cell
wall
 Spheroplasts: derived from Gram-negative
bacteria; produced artificially by lysozyme or by
penicillin or any other agent capable of breaking
down the peptidoglycan layer,
 Other components of PG such as Lipid A, and
LPS are intact
3. Cellular Element Enclosed within the Cell Envelope

• Contents of the Cell Cytoplasm

– Gelatinous solution
– Site for many biochemical and synthetic activities
– 70%-80% water
– Also contains larger, discrete cell masses (chromatin body, ribosomes,
granules, and actin strands)
– Location of growth, metabolism, and replication
Cytoskeleton
 Cellular "scaffolding" or
"skeleton" within the
cytoplasm.

 Major advance in prokaryotic

cell biology in the last decade
has been discovery of the
prokaryotic cytoskeleton.

 Up until recently, thought to be

a feature only of eukaryotic
cells.
Mesosomes: are complex
invaginations of cytoplasmic
membrane into the cytoplasm seen
in many bacteria, but not in all.
• Increase in the total surface area of the
membrane.
• Mesosomes are attached to
chromosomes and are involved in DNA
segregation during cell division .
• It is involved in respiratory enzyme -
activity. (Site of oxidative
phosphorylation)
 Bacterial Chromosome
• It is not contained in a structure like a true
chromosome –
• There is no membrane round it and it is merely
one long single strand or loop of nucleic acid
(DNA), somewhat spread out in the cell
cytoplasm as a nucleoid.
• Carries the genetic information needed for
producing several thousand enzymes and other
proteins
Plasmids
• Nonessential, circles of DNA (5-100 genes)
• Present in cytoplasm but may become
incorporated into the chromosomal DNA
• Often confer protective traits such as drug
resistance or the production of toxins and
enzymes
• Pass on in conjugation
 Inclusions bodies
– Some bacteria lay down nutrients in these
inclusions during periods of nutrient abundance
– Serve as a store house when nutrients become
depleted
– Some enclose condensed, energy-rich organic
substances
– Some aquatic bacterial inclusions include gas
vesicles to provide buoyancy and flotation
Ribosomes
 Found within cytoplasm or
attached to plasma
membrane.

 Made of protein & rRNA.

 Cell may contain

thousands

 Protein synthesis
 Cytoplasmic Granules
• A type of inclusion body
• Can store a variety of molecules, including nutrients, energy, and waste
products
• Are not enclosed by membranes
• Staining of some granules aids in identification.

Figure 4.19
Bacterial Endospores: An Extremely Resistant Stage
• Dormant, tough, non-reproductive
structure produced by small number
of bacteria.

• Resistant to radiation, desiccation,

lysozyme, temperature, starvation,
and chemical disinfectants.

• Endospores are commonly found in

soil and water, where they may
survive for very long periods of time.
Figure 4.1
 Bacteria reproduction
• Binary fission - this process involves copying the chromosome and
separating one cell into two
– asexual form of reproduction
• Transformation - the prokaryote takes in DNA found in its environment that
is shed by other prokaryotes.
• Transduction - bacteriophages, the viruses that infect bacteria, sometimes
also move short pieces of chromosomal DNA from one bacterium to another
• Conjugation - DNA is transferred from one prokaryote to another by means
of a pilus