Morphology of Bacteria

Depending on their shape, bacteria are classified into several types:

1. Cocci (from kokkos, meaning berry): These are oval or spherical cells. These cocci may be
arranged in pairs (diplococci), chains (streptococci), clusters (staphylococci) and groups of
four (tetrads) or eight (sarcina).
2. Bacilli (bacillus, meaning rod): These are rod shaped cells. Some of these bacilli may be
having peculiar arrangement or shape as follows:
(i) Cocco bacilli- length of bacteria is approximately same as its width e.g. Brucella.
(ii) Streptobacilli- These bacilli are arranged in chains e.g. Streptobacillus.
(iii) Chinese letter or cuneiform pattern-arranged at angles to each other e.g.
Corynebacterium.
(iv) Comma-shaped- curved appearance e.g. Vibrio.
(v) Spirilla- rigid spiral forms e.g. Spirillum.
3. Spirochaetes (from spiera meaning coil; chaite meaning hair): These are slender, flexous
spiral forms e.g. Treponema.
4. Actinomycetes (from actis, meaning ray, mykes meaning fungus): These are branching
filamentous bacteria resembling fungi. They have a rigid cell wall.
5. Mycoplasmas: These bacteria are cell wall deficient and hence do not possess a stable
shape. They may occur as round or oval bodies and as interlacing filaments. They are very
small in size (50-300 nm in diameter). They can reproduce in cell-free medium.
6. Rickettsiae and Chlamydiae: These are very small, obligate parasites. Due to their inability
to grow outside living cells, they were previously considered as viruses. Now they are
classified as bacteria because of typical bacterial cell wall, possession of various bacterial
enzymes and structural similarities with bacteria.

Bacterial anatomy
The outer layer or cell envelope of a bacterial cell consists of two components
1. a rigid cell wall.
2. underlying cytoplasmic or plasma membrane.
The cell envelope encloses the protoplasm which comprises cytoplasm, cytoplasmic
inclusions (mesosomes, ribosomes, inclusion granules, vacuoles) and a single circular
chromosome of deoxyribonucleic acid (DNA) . Besides these essential components, some
bacteria may possess additional structures, such as, capsule, flagella and fimbriae.
1. THE CELL WALL
The cell wall is a tough and rigid structure, surrounding the bacterium like a shell. It
weighs about 20-25% of the dry weight of the cell. The thickness of Gram negative
cell wall is 10-25 nm. The cell wall has following functions:
1. Accounts for the shape of the cell.
2. Provides protection to the cell against osmotic damage.
3. Confers rigidity upon bacteria.
 4. It takes part in cell division.
5. It possesses target site for antibiotics, lysozymes and bacteriophages. It carries
bacterial antigens that are important in virulence and immunity.
The rigid part of the cell wall is a peptidoglycan which is a mucopeptide (murein)
composed of N-acetyl murarnic acid and N-acetyl glucosamine molecules alternating
in chains, cross linked by peptide subunits
Gram-negative Cell Wall
The Gram negative cell wall is a complex structure with the following
components. (a) Lipoprotein layer: It connects the peptidoglycan to outer
membrane.
(b) Outer membrane: this contains certain proteins named as outer membrane
proteins (OMP). These are target sites for phages, antibiotics and bacteriocins.
(c) Lipopolysaccharide (LPS): This layer consists of lipid A, to which is attached
a polysaccharide. LPS constitutes the endotoxin of gram-negative bacteria. The
polysaccharide determines a major surface antigen, the O antigen. The toxicity
(pyrogenicity, lethal effect, tissue necrosis) of bacteria is associated with lipid A.
(d) The periplasmic space: It is the space in between the inner and outer
membranes. It contains various binding proteins for specific substrates.
Peptidoglycan.
ii) Gram-positive Cell Wall
Peptidoglycan: This layer in Gram positive bacteria is thicker (16-80 nm) than that
in Gram negative bacteria (2 nm).
(b) Teichoic acid: Gram positive cell wall contains significant amount of teichoic
acid which is absent in Gram negative bacteria. The teichoic acids constitute
major surface antigens of Gram positive bacteria. They are water soluble
polymers, containing ribitol or glycerol polymers. Teichoic acids are of two types,
cell wall teichoic acid and membrane teichoic acid. The cell wall teichoic acid is
covalently linked to peptidoglycan; and the membrane teichoic acid to
cytoplasmic membrane.
(c) Other components: Certain Gram positive cells also contain antigens such as
protein and polysaccharides.

Bacteria with defective cell wall

 Bacteria with defective cell wall may probably have a role in the persistence of
certain chronic infections such as pyelonephritis. Bacteria without cell walls or
with deficient cell walls are of four types:
Mycoplasma: This is a naturally occurring bacteria without cell walls.
Mycoplasma is classified as an independent bacterial genus. They don't require
hypertonic environment for maintenance and are stable in culture medium.
L -forms: Kleineberger-Nobel, while studying Streptobacillus moniliformis in the
Lister Institute, London, observed abnormal forms of the bacteria and named them
L-forms after the Lister Institute. L-forms develop either spontaneously or in the
presence of penicillin or other agents that interfere with synthesis of cell wall. L-
forms are sometimes spontaneously formed in patients treated with penicillin. L-
forms are more stable than protoplasts and spheroplasts.
Protoplasts: These are derived from Gram positive bacteria. They contain
cytoplasmic membrane and cell wall is totally lacking. These are produced
artificially by lysozyme in a hypertonic medium. These are unstable. Hypertonic
condition is necessary for their maintenance.
Spheroplasts: These are derived from Gram negative bacteria. They are produced
in presence of penicillin. They are osmotically fragile and must be maintained in
hypertonic culture medium. They differ from the protoplast in that some cell wall
material is retained.

2 Cytoplasmic membranes
It is 5-10 nm thick elastic semipermeable layer which lies beneath the cell wall
separating it from the cell cytoplasm. Electron microscopy shows the presence of
three layers constituting a unit membrane. Chemically, it consists of lipids and
protein molecules. Sterols are absent, except in mycoplasma. Cytoplasmic
membrane act as an osmotic barrier. It is the site of numerous enzymes (permease,
oxidase, polymerase) involved in the active transport of selective nutrients. It acts
as a semipermeable membrane controlling the inflow and outflow of metabolites
to and from the protoplasm. It also contains cytochrome oxidase, enzymes of
tricarboxylic acid cycle and enzymes necessary for the cell wall synthesis.

3. CYTOPLASM
The bacterial cytoplasm is a colloidal system containing a variety of organic and
inorganic solutes in a viscous watery solution. It lacks mitochondria and
endoplasmic reticulum of eukaryotic cell. It contains ribosomes, mesosomes,
vacuoles and inclusions. The cytoplasm stains uniformly with basic dyes in young
cultures.
(i) Ribosomes These are the centres of protein synthesis. These are
composied of ribosomal RNA (rRNA) and ribosomal proteins. Ribosomes
are integrated in linear strands of mRNA to form polysomes and it is at
this site that code of mRNA is translated into peptide sequences. They
arel0-20 nm in size with a sedimentation constant of 70 S (S for Svedberg
units). Each 70 S unit consists of a 30 S and a 50 S subunit.
(ii) Intracytoplasmic inclusions These are sources of stored energy and present
in some species of bacteria. They are most frequent in bacteria grown
 under conditions of nutritional deficiency and disappear when the deficient
nutrients are supplied.
(iii) Mesosomes (chondroids) Mesosomes are vesicular, convoluted structures
formed as invaginations of the plasma membrane into the cytoplasm. They
are the principal centres of respiratory enzyme and are analogous to
mitochondria of eukaryotes. These are more prominent in Gram positive
bacteria.

4. NUCLEUS
Bacterial nucleus has no nuclear membrane or nucleolus. It cannot be
demonstrated by ordinary microscope but needs an electron microscope. The
nuclear deoxyribonucleic acid (DNA) doesn't appear to contain any basic protein.
The genomic DNA is double stranded in the form of a circle. It measures about 1
mm (1000 µm) when straightened. The bacterial DNA is haploid, replicates by
simple fission and maintains bacterial genetic characteristics.
Some bacteria may possess extranuclear genetic material in the cytoplasm
consisting of DNA named as plasmids or episomes. The plasmid replicates
autonomously. They are not essential for the life of the cell, but may confer on the
bacteria certain properties, such as drug resistance and toxigenicity which
constitute a survival advantage to the bacteria. These plasmids can be transmitted
from one bacterium to another either by conjugation or by the agency of
bacteriophage. Besides these methods, plasmids may be transferred to daughter
cells during cell division by binary fission.
5. BACTERIAL CAPSULE AND SLIME LAYER
It is amorphous viscid bacterial secretions which surrounds some bacteria as their
outermost layer (Fig. 2.2). When it diffuses into the surrounding medium and
remains as a loose undermarketed secretion as in Leuconostoc, it is known as
slime layer. When this secretion is organised into a sharply defined structure, as in
Streptococcus pneumoniae (pneumococcus), it is known as the capsule. Capsules
which are very thin and cannot be demonstrated under the light microscope are
ca11ed microcapsules e.g. Nezsseria meningitidis. The slime layer or capsule is
generally-polysacchanae in nature but it is polypeptide in anthrax bacillus. Some
bacteria may have both a capsule and a slime layer (e.g. Streptococcus salivarius).
Bacteria like Klebsiella secreting large amount of slime produces mucoid growth
on agar, with a stringy consistency when touched with the loop.
Functions
(i) Capsule enhances bacterial virulence by inhibiting phagocytosis. Loss of
capsule may render the bacterium avirulent. Bacteria tend to lose capsules
on repeated subcultures.
(ii) It acts as protective covering against antibacterial substances such as,
bacteriophages, phagocytes, enzymes.
(iii) Capsular antigen is specific for bacteria and can be used for identification
and typing of bacteria.
Demonstration of Capsule: Capsule has little affinity for basic dyes; therefore, it
can't be stained by Gram staining. The following methods have been used for
demonstration of capsule.
Indian ink staining (negative staining): Capsule appears as a clear halo around
bacterium as the ink can't penetrate the capsule.
Serological methods: Capsular material is antigenic and can be demonstrated by
mixing it with a specific anticapsular serum. When a suspension of capsulated
bacterium is mixed with its specific anticapsular serum and examined under the
microscope, the capsule appears 'swollen' due to increase in its refractivity. This
phenomenon &called as capsule swelling reaction or Quelling phenomenon. This
is also named as Neufeld reaction after the person who described it. This was
widely employed for the typing of pneumococci.
Special capsule staining: These techniques employ copper salts as mordants for
staining of capsule.
FLAGELLA : Flagella are cytoplasmic appendages protruding through cell wall.
These are thread-like structures composed of a protein (flagellin), 5-20 µm in
length and 0.01-0.02 µm in diameter. They are organ of locomotion. All motile
bacteria, except spirochaetes, possess one or more flagella.
Parts and Composition Each flagellum consists of three parts
(i) filament
(ii) hook
(iii) basal body
The filament lies external to the cell and is connected to the hook at the cell
surface. The hook-basal body portion is embedded in the cell envelope. The basal
body contains outer and inner rings by which the basal body is attached to the
cytoplasmic membrane. Outer rings are absent in Gram positive bacteria. The
flagella is made up of protein...(flagellin) which is similar to myosin. Although
chemical composition of deferent genera of bacteria is similar, they are
antigenically.
Arrangement/Types:
Monotrichous- Single polar flagellum (at one end) e.g. Vibrio cholerae.
Amphitrichous-Single flagellum at both the ends e.g. Alcaligenes faecalis.
Lophotrichous-Tuft of flagella at one or both ends e.g. Spirilla.
Peritrichous: flagella arrange all around the cell e.g salmonella typhi
Demonstration: Flagella are about 0.02 µmin thickness and hence beyond the
resolution limit of the light microscope. The following methods are used for its
demonstration:
(i) Dark ground illumination.
(ii) Special staining techniques in which thickness of flagella is increased by
mordanting.
(iii) Electron microscopy
9

FIMBRIAE: These are hair-like. appendages projecting from the cell surface as
straight filaments. They are also called pili. They are 0.1 to 1.0 µm in length and
less than 10 nm thick (shorter and thinner than flagella). Fimbriae are found in
some Gram negative bacteria. Each bacterium possesses 100-500 fimbriae
peritrichously. They are more numerous than flagella. They are antigenic. They
tend to disappear when subcultures are made on solid media. Fimbria is composed
of protein called pilin. They are unrelated to motility and are found on motile as
well as non-motile bacteria.
Types There are three main types of fimbriae.
Common pili- These are of six types depending / on their morphology, number per
cell, adhesive properties and antigenic nature.
Sex or F (fertility) pili
Col I ( colicin) pili
Functions
(i) Adhesion- Fimbriae are organs of adhesion. This property enhances the
virulence of bacteria.
(ii) Transfer of genetic material: sex pili are present in male bacteria. They
help the male cells to attach with non-male (female) cells in forming
"conjugation tubes" through which genetic material is believed to be
transferred from the donor (male) to the recipient (female) cell.
Detection of Fimbriae
(i) Electron microscopy.
(ii) Haemagglutination- Many fimbriated bacteria ( e.g. escherichia coli,
Klebsiella) strongly agglutinate red blood cells of guinea pigs, fowl, horses and
pigs; human and sheep cell ls weakly and ox cells scarcely. This property of
haemagglutination is a simple method for detecting the presence of fimbriae. The
haemagglutination can be specifically inhibited by D-mannose.
Bacterial spores
Spores are highly resistant resting stage formed in unfavourable environmental
conditions presumed to be related to the depletion of exogenous nutrients. As
bacterial spores are formed within the parent cell, these are called endospores.
Sporulation is not a method of reproduction. In the process of sporulation, each
vegetative cell forms only one spore and during subsequent germination, each
spore gives rise to only one vegetative bacterium.
Sporulation and morphology of spores
Bacterial cell undergoes spore formation in nutritionally deprived conditions and
this process is called sporulation. Spore develops from a portion of protoplasm
(forespore) near one end of the cell. The remaining part of cell is called
sporangium. Bacterial DNA replicates and divides into two DNA molecules. One
of these is incorporated into forespore and other into sporangium. A transverse
septum grows across the cell from the cell membrane. It divides forespore and
sporangium. The forespore is completely encircled by this septum as a double
layered membrane. The inner layer becomes the spore membrane and the outer
layer becomes thickened spore coat. Between the two layers is spore cortex.
Some spores have an additional apparently rather loose, outer covering called
exosporium.
Shape and Position of Spores:
The precise position, shape and relative size of spore are constant within a
particular species. Spores may be central, subterminal or terminal. They may be
oval or spherical in shape. The diameter of spore may be same or less than the
width of bacteria (Bacillus), or may be wider than the bacillary body producing a
distension or bulge in the cell (Clostridiufn ).
Resistance
Bacterial spores are extremely resistant to ordinary boiling, disinfectants and
heating. The high resistance of spores is due to hi content of calcium and
dipicolinic acid; low water content; the thick impervious cortex and spore coats;
their low metabolic and enzymatic activity. However, spores of all medically
important bacteria are destroyed by autoclaving at 121 °C for 15 minutes.
Germination
The process of conversion of a spore into vegetative cell under suitable
conditions is known as germination. The spore loses its refractivity, and swells
when transferred to conditions conducive to vegetative growth. The spore wall is
shed and the germ cell appears by rupturing the spore coat. The formation of
vegetative bacterium occurs by elongation of the germ cell.
Demonstration: Gram staining Spores appear as an unstained refractile body
within the cell.
Modified ·Ziehl-Neelsen (ZN) staining- Spores appear as acid-fast (red colour).
Ziehl-Neelsen staining with 0.25-0.5% sulphuric acid (instead of 20% sulphuric
acid as used in conventional method) as decolouring agent is used spore staining.
Uses of Spores: Spore of certain species of bacteria are employes as indicator for
proper sterilization e.g. Bacillus stearothermophilus which is destroyed at a
temperature of 121 °C for 10-20 minutes (same temperature and time as used in
autoclaving). These spores may be kept in autoclave prior to its use. Absence of
the spores after autoclaving indicates proper sterilisation.
Spore Forming Bacteria Obligate aerobes- genus Bacillus e.g. B. anthracis and B.
subtilis Obligate anaerobes- genus Clostridia e. g. Cl. tetani, Cl. welchii, Cl.
botulinum
Both Bacillus and Clostridia bacilli.