INTRODUCTION TO

BACTERIOLOGY
DAVID K. NGUNJIRI
 BACTERIOLOGY
• STUDY OF BACTERIA
• Learning objectives
By the end of the lesson you will be able to
1. Define a bacteria
2. Describe the general structure of a bacteria cell as
seen under a light microscope.
3. Describe the unique morphological features of certain
bacteria and
1. state function of these features indicating the examples of
bacteria that possess them.
 INTRODUCTION TO BACTERIA
• Bacteria are the large group of unicellular microorganisms that are ubiquitous in
nature.
• They are extremely small organisms and cannot be seen by the naked eyes.
• On the solid media plates, they grow as the colonies that contain the large no.
of individual cells of the bacteria.
• A bacteria is a single cell organism (unicellular)
• An individual bacteria cell measures 0.5- 1cm in width and 0.5 – 8cm in length
• Bacteria are the Prokaryotic organisms characterized by the lack of the well-
defined nucleus, cell-organelles like Mitochondria, Golgi apparatus, Endoplasmic
Reticulum etc.
• contains 70S Ribosome (30S & 50S Subunits), surrounded by a protective cell
wall containing peptidoglycan, have small rings of DNA called Plasmids.
 Basic structure
• A bacteria cell is generally like any other cell thus most of the structure found
in ordinary cells are found in bacteria.
• Observed under light microscope shows the following structures;
1. An outer layer or cell envelope ;consists of two components:
i. Cell wall (cell wall is rigid)
ii. Cytoplasmic Membrane (or plasma membrane below the cell wall)
2. Cytoplasm
• Inside the cell envelope, there is protoplasm consists of the cytoplasm and
cytoplasmic inclusions such as;
– Ribosomes,
– mesosomes,
– granules,
– vacuoles and
– the nuclear body.
A TYPICAL BACTERIAL CELL
 BACTERIA CELL WALL
• It is a tough outer and rigid structure
• It surrounds the bacteria like a shell
• gives a particular shape to a bacterium
• It is rigid & permeable .
• prevents swelling and bursting of the cell.
• Without the wall, the cell will burst if placed in hypotonic media.
• It protects the internal structures of the bacterial cell.
•  It is about 10 – 25nm in thickness.
•  It weighs about 20 – 30% of the dry weight of the cell.
•  Chemically the cell wall is composed of peptidoglycan which is composed of
two amino acids alternatively arranged and cross-linked by a peptide bond.
i. N-acetyl glucosamine (NAG) and
ii. N-acetyl muramic acid (NAM)
 peptidoglycan layer
• It is present in both gram-positive and gram-negative organisms.
• It is composed of repeating disaccharides with 4 amino acids in a
side chain extending from each disaccharide.
• The amino-acid chains of the peptidoglycan covalently bind to
other amino acids from neighboring chains resulting in a stable
cross-linked structure.
• The enzyme that catalyzes the formation of this linkage is called
transpeptidase and is located in the inner cytoplasmic
membrane.
• penicillin binds to and inhibits this enzyme,hence it is also called
penicillin binding protein.
 chemical nature of bacterial cell wall
•  Due to differences in the chemical nature of the
bacterial cell wall, bacteria differ in staining property.
i.  Gram-negative bacteria contain lipopolysaccharide,
ii. Gram-positive bacteria contains teichoic acid in cell wall
iii. Acid-fast bacilli contain mycolic acid in the cell wall.
NB//
•  The cell walls of gram-positive bacteria have simpler
chemical nature than those of gram-negative bacteria.
• Animal cells have only a single cytoplasmic membrane
composed of a phospholipid bilayer.
 Cell wall
The gram-positive cell wall.
• very thick and has extensive cross-linking of
the amino-acid side chains.
The gram-negative cell wall ;
• very thin with a fairly simple cross-linking
pattern.
 CYTOPLASMIC/PLASMA MEMBRANE OF
BACTERIA:
• This is the inner membrane lying next to the cell wall.
• It is a thin and semi permeable membrane;
•  It is 5-10nm thin
• Chemically, the membrane a phospholipid bilayer consisting
of lipoproteins with small amounts of carbohydrates.
NB//
• It is a phospholipid bilayer such as found in the membrane of
mammalian cells.
• But unlike mammalian & fungi cells, it rarely contains sterols
 chemical composition
• similar to that of membranes in mammalian cell:
phospholipid
– 20-30% lipid
– protein 60%.
NB //
1. Unlike animals, the bacterial cytoplasm membrane has no
cholesterol or other sterols.
2. Teichoic acid is an important polysaccharide present in the
gram positive cell wall.
3. It acts as an antigenic determinant, hence important for
serologic identification of many gram-positive species. 
 Cytoplasmic membrane functions
• It acts as a semi-permeable membrane –
controls the inflow and outflow of metabolites
to and from the cytoplasm. i.e.It determines
which substances leave or enter the cell
• Permeable to all ions.
• non-ionized molecules larger than glycerol
penetrate very slowly except by specific
transport.
 functions of the membrane
• It contains enzymes necessary for cell wall synthesis.
• Also contains oxidative enzymes such as
– cytochromes,
– quinones,
– ATPase
• Membrane contains selective transport systems to pump in
nutrients from dilute external media .eg
– specific sugars,
– amino acids,
– metals, etc
• Simple passive diffusion is insufficient for the bacterial cell in its
usual environment of dilute nutrients.
 CYTOPLASM OF BACTERIA
•  It is the liquid part of the cell.
• It is a viscous watery solution containing a
variety of organic and inorganic solutes.
• It differs from the eukaryotic cytoplasm in
– not exhibiting internal mobility (protoplasmic
streaming)
– the absence of endoplasmic reticulum or
mitochondria.
 Cytoplasm composition
• It contains the essential component of the cell
such as
i. the nuclear
ii. Ribosomes,
iii. Mesosomes,
iv. Intracytoplasmic inclusions, and vacuoles.
 NUCLEUS IN BACTERIA
•  Bacteria do not possess a true nucleus.
• It has no nuclear membrane or nucleolus
• no organization of DNA into visible
chromosomes.
• Nucleoid which is the nuclear equivalent is a
single large molecule of double stranded DNA
which is coiled and resides freely inside the
cytoplasm.
 Bacterial DNA
• DNA molecules are necessary for reproduction
& the antigenic typing of the bacteria.
• consists of a single circle of double-stranded
DNA.
• Some bacteria possess extra Smaller adjacent
circles of double-stranded DNA (chromosomal
genetic material) called plasmids or episomes.
• they often contain antibiotic resistance genes.
 Mesosomes
• Mesosomes are vesicular, multi-laminated or
convoluted structures formed as invaginations
of the plasma membrane into the cytoplasm.
• Principal sites of respiratory enzymes in bacteria
• They are analogous to mitochondria in
eukaryotes cells.
• More prominent in gram-positive bacteria.
• Also called Chondroid
 2. Ribosomes ;
• Ribosomes are the sites of protein synthesis.
• composed of protein and RNA involved in the
translation process, during the synthesis of proteins.
• Bacteria Ribosomes have a sedimentation constant
70S; made up of 50S and 30S subunits.
• NB//Bacteria as procaryotes, have smaller ribosomes
(70S) than animals eucaryotes (80S)
 Bacterial ribosomes subunits
• Bacterial ribosomes consist of 2 subunits
1. a large subunit (50S)
2. a small subunit (30S).
• These numbers relate to the rate of sedimentation.
• Antibiotics have been developed to attack like magic
bullets attack by inhibiting protein synthesis at the
bacterial ribosomal subunits while leaving the
animal ribosomes alone.
– Erythromycin works at the 50S subunit.
– Tetracycline blocks protein synthesis at the 30S subunit.
 Intra cytoplasmic inclusions
• These are the storage form of food material in
bacteria which may be of various types,
• The chief of which are ;
– volutin,
– polysaccharide,
– lipid and
– sulfur granules.
• These are characteristic of different species and
depend on the age and condition of the culture.
NTRACYTOPLASMIC INCLUSIONS
inclusions Other names Found in
Volutin granules Polyphosphate/ Corynebacterium,
metachromatic/ babes- spirillum, rhizobium and
Ernst granules bacillus.
Polysaccharides Polyglucan or Clostridium,
Iodophillic granules Citrobector, Escherichia,
klebsiella
Lipid granules Sudanophillic or poly- Rhizobium, bacillus,
beta-hydroxy butyrate alcaligenes and soil
granules bacteria
Sulfur granules Sulfur Globules Thiomagrita,
Thiobacillus.
 Vacuoles –
• These are fluid-filled cavity present in the
bacteria.
• Separated from the cytoplasm by a
membrane.
• Gas vacuoles are gas-filled cavities present in
bacteria and provide buoyancy to organisms in
the aquatic habitat.
DIFFERENCES BETWEEN GRAM-
POSITIVE AND GRAM-NEGATIVE
ORGANISMS
 The gram-positive cell envelope
• Has an outer cell wall composed of complex
cross-linked peptidoglycan, teichoic acid,
polysaccharides and other proteins.
• The inner surface of the cell wall touches the
cytoplasmic membrane.
• The cytoplasmic membrane contains proteins
that span the lipid bilayer.
Gram Positive
 gram positive layer
• Thick peptidoglycan cell wall structure, with
intertwined chains of teichoic acid emerging at the
cell surface .
• Organization of underlying cytoplasmic membrane
shows bilayer phospholipids, transmembrane and
inner and outer leaflet proteins,
• transmembrane proteins involved in active
transport, and proton channel (the energy-
transducing system).
 The gram-negative cell envelope.
• Gram-negative bacteria has 3 layers,excluding
the periplasmic space.
 1) a cytoplasmic membrane
2) a peptidoglycan layer.
3) outer cell membrane.
the gram-negative cell envelope.
  1) a cytoplasmic membrane
• The inner cytoplasmic membrane as in gram-
positive bacteria contains a phospholipid bilayer
with embedded proteins .
• a periplasmic space
• There is a periplasmic space between the
cytoplasmic membrane and peptidoglycan layer.
• This periplasmic space is filled with a gel that
contains proteins and enzymes.
 2) a peptidoglycan layer.
• An extremely thin layer
• Does not contain teichoic acid.
• have a small helicallipoprotein called murein
lipoprotein.
• This lipoprotein originates from the
peptidoglycan layer and extends outward to
bind the third outer membrane
 3) outer cell membrane.
• This last membrane is similar to other cell
membranes.
• in that it is composed of two layers of
phospholipid (bilayer) with hydrophobic tails
in the center.
• 1) oligosaccharide -Outer carbohydrate chains
• 2) The center part
• 3) lipid A, Interior to the core
 i) Outer oligosaccharide carbohydrate

• Made of chains of 1-50 oligosaccharide units

that extend into the surrounding media.
• These differ from one organism to another
and are antigenic determinants.
• This part is called the 0-specific side chain or
the 0-antigen.
• 2) The center part is a water soluble core
polysaccharide.
 lipid A
• 3) Interior to the core polysaccharide is the
third component, lipid A,
• It is a disaccharide with multiple fatty acid
tails reaching into the membrane.
• Lipid A is toxic to humans and is known as the
gram-negative endotoxin.
 porins
• Embedded in the gram-negative outer
membrane are porin proteins, which allow
passage of nutrients.
• These are also unique to gram-negative
organisms.
 DIFFERENCES BETWEEN GRAM-
-POSITIVE AND GRAM-NEGATIVE ORGANISMS
Gram-Positive Cells
Low lipid content
2 Layers:
1. Inner cytoplasmic membrane
2. Outer thick peptidoglycan layer
(60-100% peptidoglycan)
NO endotoxin (except Listeria
monocytogenes)
NO porin channel
Vulnerable to lysozyme and
penicillin attack
Gram-Negative Cells
High lipid content
3 Layers:
1. Inner cytoplasmic membrane
2. Thin peptidoglycan layer
(5- 10% peptidoglycan)
3. Outer membrane with
lipopolysaccharide (LPS)
Endotoxin (LPS) - lipid A
NO periplasmic space
Porin channel
Resistant to lysozyme and
penicillin attack
 clinical significance
• The differences between gram-positive and gram negative
organisms result in varied interactions with the environment.
The gram-positives
• have a thick meshed peptidoglycan layer
• Allows the diffusion of low molecular weight compounds,
• so substances that damage the cytoplasmic membrane such as
antibiotics, dyes,and detergents can easily pass through.
the gram negative
• outer lipopolysaccharide cell membrane blocks the passage of
substances such as antibiotics, dyes,and detergents .
• Therefore, antibiotics and chemicals that attempt to attack the
peptidoglycan cell wall such as penicillins and lysozyme are
unable to pass through.
 SPECIAL MORPHOLOGICAL
CHARACTERISTICS OF BACTERIA
• Bacteria has also been able to acquire certain
structure which are not common to all
bacteria
• These includes
i. capsules,
ii. spores
iii. flagella and
iv. cilia (fimbriae or pili
 CAPSULE
• Many bacteria are surrounded by discrete
covering layer of relatively firm gelatinous
material that lies outside and in contact with
the cell wall in wet condition.
– NB/ Gelatinous refer to viscosity of the surrounding.
• It is an amorphous viscid secretion which
surrounds them as the outermost layer.
–  When it remains as loose un-demarcated secretion,
it is known as the slime layer.
– When this secretion is organized into a sharply
defined structure, it is known as the capsule.
 CAPSULE
• Capsules are protective walls that surround the
cell membranes.
• Both g-positive and g-negative bacteria can form

• Formed to resist phagocytosis by the tissue cell

• Enable bacteria to be more virulent because
macrophages and neutrophils are unable to
phagocytize the encapsulated bugs.
• .
 Capsular development
• Capsular development is determined by the environment
conditions
• It is best developed when organism is living in growing tissue.
• Bacteria secrete these sugar moieties, which then coat their
outer wall.
• They may be formed by
1. carbohydrates polysaccharides or
2. proteins polypeptides
3. Bacillus anthracis,is unique in that its capsule is made up of amino acid
residues.
• NB//Polysaccharides formed capsules are more resistance to
heat than polypeptides
 PURPOSE OF CAPSULES
1. They protect them against
– Lytic enzymes
– Phagocytosis by white blood cells
– Adverse variation in the temperature
2. Capsules enhance the;
1. invasiveness and
2. virulence of bacteria.
3. The capsule substance is also antigenic and
– capsular antigen plays an important role in determining the antigenic
properties of bacteria

NB//Capsules are not meant for any reproduction

For example,Streptococcus pneumoniae has
a capsule
 bacteria with capsules
• Examples of bacteria which posses capsules
include:-
1. Streptococcus pneumonia
2. Streptococcus pyogenes
3. Klebsiella pneumonia
4. Bacillus anthracis
5. Yersinia pestis
 
 SPORES
• Some bacteria have the ability to form the highly resistant resting
stage during unfavorable environmental conditions called as
Spores.
• spores are metabolically dormant forms of bacteria that are
resistant to
– heat (boiling),
– cold, drying and
– chemical agents.
• Since the spores are formed inside the bacterial cell, are called as
endospores.
• Sporulation is the process by which bacterial cell forms spore
under unfavorable conditions.
• Such bacteria are called spore bearing or sporing
bacteria
• The process of spore formation is called
sporulation
• Sporulation occurs when the environmental
condition do not favor the normal bacteria growth.
E.g. in starvation- adverse severe condition
• Bacteria severe are endospores and are not used
for reproduction.
 SPORES

• Spores form when there is a shortage of needed

nutrients and can lie dormant for years.
• Surgical instruments are heated in an autoclave,
which uses steam under pressure, to 121°C for 15
minutes, in order to ensure the destruction of
Clostridium and Bacillus spores.
• When the spore is exposed to a favorable
nutrient or environment,
• it becomes active again.
 Spore development
•  Spore develops from a part of protoplasm is called
as forespore near one end of the cell and the
remaining part of the cell is called sporangium.
• They have a multilayered protective coat consisting
of:
1. A cell membrane
2. A thick peptidoglycan mesh
3. Another cell membrane
4. A wall of keratin-like protein
5. An outer layer called the exosporium
 Spores characteristics
• Bacterial spores are highly resistant to
– ordinary boiling,
– disinfectants, and
– beating.
•  Germination is the process of conversion of bacterial spores into a
vegetative cell under suitable conditions.
• NB/ Spores are formed during adverse condition while capsules
are formed to enhance invasiveness and virulence of the bacteria
when inside a living tissue
• None of them are involved in reproduction
• Exopores are formed by fungi and these may be involved in
reproduction
 spores forming bacteria
• Examples of the bacteria that sporulate are
1. Bacillus anthracis
2.  Bacillus stearothermophillus
3. Clostridium tetani
4. Clostidium welchi
5. Clostidium botulinum
6.  Clostridium perfringes 
size of the spore relative to the bacteria

• The size of the spore relative to the bacteria

may be described as follows
• Bulging – when they cause bulging of the cell
wall
• Non bulging – when they don’t cause bulging
of cell wall
  Shape of spores –
• Shapes –
i. Oval or
ii. Spherical
 STRUCTURE AND SITUATION OF BACTERIA
SPORES
• The bacteria spores may be
• Oval
• Spherical
• They may be situated
• Centrally
• Sub-terminally
• Terminally
Spherical and central
Oval and central
 Bulging
1. central spore

2.sub-terminal spore

3.Terminal spore
 Position
Non- bulging spores –

–
i. central

ii sub-terminal

Iii terminal
 FLAGELLA OF BACTERIA
• ⇒ Motile bacteria possess one or more
unbranched, long sinuous filaments called
flagella, are cytoplasmic appendages
protruding from the cell wall.
• ⇒ They are the organ of locomotion in the
bacterium.
• ⇒ Composed of a protein – flagellin, 5-20mm
in length and 0.02mm in
 FLAGELIAE
• They are long filamentous appendages
• They are about 0.02 in diameter
• They range from 1-20 par bacteria
• They are in ration of 1:15 to the body length
• May be situated in characteristics sites such as
• polarize originating from one or both ends
• Lateral originating from the sides
 Function of flagellae
• To facilitate the mobility of the bacteria
• Believed to increase the rate of the nutrients solute
intakes by continuous change of environment fluid in
contact with the bacteria cell
• Ensuring that at least some cell reach all locality suitable
for colonization by random movement and dispersion.
• Moving toward areas of favorable conditions and away
from areas of disinfectant and antibiotics .
• This is because bacteria prefer to grow in areas of high
nutrient and do not do well in adverse conditions.
 Arrangement of flagella
• ⇒ Arrangement of flagella varies species to
species,
1. Monotrichous – Presence of one flagellum at one
pole, e.g.
2. Amphitrichous – Presence of tufts of flagella at both
ends, e.g. Spirillum.
3. Lophotrichous – Presence of tuft of flagella at one
pol, e.g.psedomonas
4. Peritrichous – Presence of many flagella surrounding
the outer surface of the body, e.g. Bacillus.
 NOTE
• Bacteria with flagella are motile
• NB/ Certain bacteria are able to move even
when they are not flagellated e.g spirochetes
• Possession of flagellum and motility of micro-
organism are significant features in identification
• Examples ..
– Entro-bacteria are classified on whether they are are
flagellated and motile or not.
 D.Fimbriae/cilia
• Some certain bacilli possess filamentous
appendages out of different kind from flagella
called fimeriac
• They are more numerous than flagellae
• They are much shorter and are about ½ as thick
also called cilia
• They can’t be seen by light microscope but are
viewed under light microscope
 
 Function
• They help the bacteria cell to adhere to body
tissue and BBCS of human and other animals
• The sex fimbriae help the bacteria in
reproduction by the transfer of D.NA from
male bacteria to female.
 FIMBRIAE IN BACTERIA
• ⇒ These are very fine hair like appendages
projecting from the surface as straight filaments.
• ⇒ They are shorter and thinner than flagella, i.e.
0.1 – 1.0mm long and less than 10nm thick.
• ⇒ Fimbriae are found in some gram-negative
bacteria.
• ⇒ There are 3 types of fimbriae have been
identified – common, ‘sex’ Pili and col I (colicin)
pili.