MICROBIOLOGY

THE BACTERIA
MICROBIOLOGY – I
DR SABREEN
OUTLINE
• Bacterial cell structure
• Bacterial forms and function
• Bacterial identification and classification
• The gram stain.

• https://www.atsu.edu/faculty/chamberlain/Website/Lects/Bacteria.h
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 MICROBIOLOGY
• The science (logos) of small (micro) life (bios).
• Study of microorganisms.
MICROORGANISM
• Living things which individually are too small to be seen with the naked eye.
• All of the following may be considered microorganisms:
• bacteria (eubacteria, archaebacterial)
• fungi (yeasts, mold )
• Protozoa
• microscopic algae
• viruses
• various parasitic worms
 BACTERIA
• Bacteria is unicellular, free-living, microscopic
microorganisms capable of performing all the
essential functions of life.
• They possess both deoxyribonucleic acid (DNA)
and Ribonucleic acid (RNA).
• Bacteria are prokaryotic microorganisms that do
not contain chlorophyll.
• They occur in water, soil, air, food, and all natural
environment.
• They can survive extremes of temperature, pH,
oxygen, and atmospheric pressure.
SIZE OF BACTERIA
• Bacteria are very small microorganisms which are visible under the
microscope.
• They are having the size range in microns.
• Bacteria are stained by staining reagents and then visualised under
high power of magnification (1000X) of compound microscope.
• An electron microscope is used for clear visualization of internal
structure of bacteria.
SHAPE OF BACTERIA
• On the basis of shape bacteria are classified as
1. Cocci
2. Bacilli
3. Vibrios
4. Spirilla
5. Spirochetes
6. Actinomycetes
7. Mycoplasma
1. COCCI
• Cocci are small, spherical or oval cells.
In greek ‘Kokkos’ means berry.
Eg: micrococcus

2. BACILLI
• They are rod shaped cells. Eg: Bacillus anthracis.
• It is derived from greek word “ Bacillus” meaning stick.
• In some of the bacilli the length of cell may be equal to width. Such
bacillary forms are known as coccobacilli.
Eg: Bracella
3. VIBRIOS
• They are comma shaped curved rods.
Eg: Vibrio comma.

4. SPIRILLA
• They are longer rigid rods with several curves or coils.
• They have a helical shape and rigid body.
• Eg: Spirillum ruprem.

5. SPIROCHETES
• They are slender and flexuous spiral forms.
6. Actinomycetes
• The characteristic shape is due to the presence of rigid cell wall.
Eg: Streptomyces.
• They are branching filamentous bacteria.
Eg: Streptomyces species.

7. Mycoplasma
• They are cell wall deficient bacteria and hence do not possess stable
morphology.
• They occur as round or oval bodies with interlacing filaments.
. ARRANGEMENT OF BACTERIAL
CELLS
• Appears as several characteristics arrangement or grouping.
1.Diplococci
2.Streptococci
3.Tetracocci
4.Staphylococci
5.Sarcinae
1. Diplococci
• They split in one plane and remains in pair.
Eg: diplococcus pneumoniae.

2. Streptococci
• These cells divide in one planes and remain attached , to
form chains.
Eg: streptococcus lactis.

3. Tetracocci
• They divide in two planes and live in groups of four.
Eg: Gaffyka tetragena.
4. Staphylococci
• Cocci cells divide in three planes in an irregular pattern.
These cells produce bunches of cocci as in grapes.
Eg: staphylococcus aureus, staphylococcus albus.

5. Sarcinae
• Sarcinae cells divide in three planes in a regular pattern.
• These cells produces a cuboidal arrangement of group of a eight
cells.
• Eg: Micrococcus tetragena
Arrangement of grouping formed by
bacilli species
1. Diplobacilli
2. Streptobacilli
3. Trichomes
 STRUCTURE OF BACTERIA

Essential components Nonessential components

• Cell wall • Capsule
Peptidoglycan
• Pilus or fimbria
Outer membrane of gram negative bacteria
Surface fibers of gram-positive bacteria • Flagellum
• Plasma membrane • Spore
• Ribosome • Plasmid
• Nucleoid
• Granule
• Mesosome
• Periplasm
• Glycocalyx
CELL WALL
• Structure of Cell Wall
• Some bacteria have surface features, external to cell wall like capsule,
flagella and pili.
• Cell wall is a multi-layered structure, located external to cell
membrane.
• It is composed of inner layer, peptidoglycan and outer layer which is
of variable thickness.
• Peptidoglycan gives structural support and maintains specific shape
of the cell.
Gram+ & Gram- Cell Wall
Composition
• Structure, chemical composition and thickness of cell wall varies in G+ and
G – bacteria.
• Peptidoglycan layer is much thicker in G+ than G- bacteria.
• Some G+ bacteria have fibers of teichoic acid, whereas G- bacteria lack it.
• In contrast, G- have complex outer layer of lipopolysachride, lipoprotein
and phospholipids.
• There is a periplasmic space between outer cell wall and cytoplasmic
membrane.
• It is the site of betalactmase enzyme which degrades penicillin and
betalactam drugs.
Properties of Cell Wall
• G- bacteria contain endotoxins (lipopolysachrides).
• Polysachrides and proteins are antigens.
• Porin proteins play role in facilitating the passage of small hydrophillic
molecules into the cell.
• They also act as channels to allow the essential substances like sugar,
aminoacids, vitamins, metals and drugs into the cell.
Cell Wall of Acid fast bacteria
• Mycobacterium tuberculosis have unusual cell wall which cannot be
stained by gram stain.
• Bacteria resist decolorization with alcohol after staining with
carbolfuchsion.
• This due to high contents of mycolic acid in the cell wall.
 Peptidoglycan
• It is complex, interwoven network which surrounds the entire cell.
• Composed of single covalently linked macro molecule present only in
bacteria to give rigid support to the cell.
• It allows the cell to withstand media of low osmotic pressure e.g water.
• It consists of peptide and sugar (glycon) which make the molecule.
• Carbohydrate is the backbone, which is composed of alternate N
acetylemuramic acid and N acetyleglusomin molecule.
• Each muramic acid molecule is attached to tetrapeptide consisting of both
D-L amino acids, composition of which differs from one bacteria to other
bacteria.
• Two important aminoacids, diaminopimelic acid and D-alanine which is
involved in cross linking of tetra peptide.
• Peptidoglycan is not present in human cells.
• It is a good target for antimicrobials.
• Drugs like penicillin, cephalosporin and vancomycin inhibit the synthesis of
peptidoglycan by preventing the transpeptidase engyme from creating cross
linkage between two adjacent tetrapeptides.
 Lysozyme
• Lysozyme is an enzyme present in human tears, saliva and mucous
which can decompose the peptidoglycan backbone by breaking its
glycerol bonds and increasing natural resistance of host against
bacteria.
• The cell swells and ruptures as a result of water entry into the cell
after treatment by lysozymes.
 Lipopolysaccharides (LPS)
• LPS of outer membrane of cell wall of G- bacteria is endotoxin,
responsible for disease features e.g fever, shock and hypotension.
• Endotoxin is the integral part of G- bacteria cell wall.
• LPS is composed of 3 distinct units:
• phospholipids called Lipid A, responsible for toxic effects.
• A core of polysachride of 5 sugars, linked through
ketodeoxyoctulonate (KDO) to lipid A.
• Outer polysachride consisting of up to 25 repeated units of 3-5
sugars.
 Teichoic Acid
• Present in G+ cell wall as fibers of glycerolphosphate/ribitolphosphate.
• Teichoic acid is linked to lipids in the cytoplasmic membrane called
lipoteichoic acid.
• Teichoic acid can induce septic shock in G+ bacteria.
Cytoplasmic membrane
• Just inside the peptidoglycan, lies the cytoplasmic membrane,
composed of phospholipid bilayer.
• It has 4 important fuctions:
• Active transport of molecules into the cells.
• Energy production by oxidative phosphorylation.
• Synthesis of cell wall presussors.
• Secretions of enzymes and toxins
MESOSOMES
• This is the invagination of cytoplasmic membrane which divides the
cell in half as the binding site of DNA, that will become genetic material
of each daughter cell.
RIBOSOMES
• It is the protein synthesis site in eukaryotics.
• Prokaryotic ribosmes (70S with 50S, 30S sub units).
• Eukaryotic ribosome (80Swith 60S, 40S sub units).
• Activity of antibiotics is dependent on robersomal RNA differences
and protein sysnthesis.
GRANULES
• Cytoplasm has different types of granules which serve as nutrient
storage that can be strained for diagnosis e.g volutin granule is the
reserve of high energy stored in the form of polymerized
metaphosphate.
• On staining, it appears as metachromatic granules (red & blue) which
are characteristics of corynebacteriumdiphtheriae, the cause of
diphtheria
 Nucleoid
• Area of cytoplasm where DNA is placed.
• Prokaryotics have DNA, which is single circular molecule weight 2 x 10 9
contains 2000 genes in comparison to human DNA with 100,000 genes.
• There is no nuclear membrane, no nucleus, no mitotic figure and no
histones.
 Plasmid

• Extra chromosomal material, double stranded circular molecule, capable of replicating

independently.
• Can be integrated into the bacterial chromosomes.
• Plasmid is present in both G+ and G- bacteria.
• Several different types of plasmids can co-exist in one cell.
• There are 2 types of Plasmids.
• Transmissible:
Can be transferred form cell to cell by conjugation process.
They are large molecules and contain dozens of genes responsible of sex pilus and enzymes
production required for transfer.
• Non Transmissible:
Small molecules and do not contain transfer genes.
FUNCTIONS OF PLASMID:
• Antibiotic resistance, mediated by enzymes.
• Resistance to heavy metals, e.g mercury used in antiseptics.
• Resistance to UV light, DNA repair enzyme.
• Resistance to pili, adherence of bacteria to cell epithelium.

MECHANISM OF PLASMIDS
• Degradation of bacterial cell membrane by making pores in the
membrane.
• Bacterial degradation of DNA by DNAse enzymes.
• Bacteriocin may be useful in treating infections by antibiotic resistant
bacteria
 Transposons
• Pieces of DNA that move from one place to another within or
between the DNAs of bacteria, plasmid and bacteriophage.
• They are also called jumping genes, they keep on making new copies
in the cytoplasm.
• They code for drug resistance enzymes, toxins or variety of metabolic
enzymes which result in mutation of genes.
• Transposons are not capable of independent replication.
• A single plasmid can contain several transposons carrying drug
resistance genes.
Specialized structures outside Cell
Wall
CAPSULE:
• Gelatinous material covering the whole bacteria.
• Composed of polysaccharide except in anthrax bacilli, which contain
polymerized D-glutamic acid.
• Sugar components vary from one bacterium to another e.g 84 different
serological types of streptococcus pneumoniae.
 Importance of Capsule:
• Determinant of virulence of many bacteria since it limits the ability of
phagocytes to engulf bacteria.
• Negative charge on capsule repels negatively charged nautrophils from eating
the bacteria.
• Specific identification of bacteria can be made by using antiserum against
polysaccharide capsule.
• The capsule will swell and this process is called quellung reaction.
• Vaccine can be made from polysaccharide capsule antigen, making specific
antibodies e.g 23 types of strept. pneumonae are present in the current
vaccine.
• Capsule plays a role in adherence of bacteria to human tissue.
• Opsonization is process by which antibodies enhance phagocytosis.
 FLAGELLA
• Long whip like structures which move the bacteria to nutrients by a process called
chemotaxis.
• These propellers are composed of many sub units of single protein flagellin, arranged in
chains.
• Energy for the movement is provided by ATP.
• Flagellated bacteria have specific number and locations on the bacterial cell wall.
• Flagella are the means of motility of the bacteria.
• Spirochetes move by flagellum like structures called axial filaments which give them
undulating movements.
• Some motile bacteria (E.colli, proteus) are common cause of UTI. Flagella can play a role in
ascending infection to urethra and bladder.
• Some bacteria (salmonella spp.) are identified in the lab by using specific antibodies
against flagellar proteins.
 Pili (Fimbriae)
• Hair like structure, shorter and straight which extends from the cell
wall.
• It is composed of pilin protein arranged in helical strands in G-
bacteria.
• It has two important roles :
• Helps in attachment of bacteria to specific receptors on human
cells. Mutants which do not have pili are non pathogenic since they
cannot anchor to the cell surface.
• Sex pilus makes the attachment between male and female bacteria
during conjugation.
 GLCOCALYX ( SLIME LAYER)
• Polysaccharide coating secreted by many bacteria.
• It forms a slimy film and allows the bacteria to adhere
firmly to various structures e.g skin, heart valves and
catheters.
• Glycocalyx has special medical importance, i.estrains of
pseudomonas aeruginosa causes respiratory tract infection
in cystic fibrosis while staph.
• Epidermidis and viridans streptococci cause endocarditis.
• Strept. mutansadheres to the teeth surface due to
glycocalyx and causes plaque formation, leading to dental
carries.
 Spores
• Highly resistant structures, formed to cope up adverse conditions
e.g G+ rods, genus bacillus (anthrax) and clostridium which includes tetnus and botulism.
• Spore formation takes place when nutrients such as carbon and nitrogen are depleted.
• The spore forms inside the cell and contains bacterial DNA, cytoplasm, cell membrane,
peptidoglycan and little water.
• Thick keratin like coat is responsible for the resistance of spores to heat, dehydration and
chemicals.
• Spores have no metabolic activity and can stay dormant for years.
• On exposure to water and necessary nutrients, the enzyme breaks the coat.
• Water and nutrients enter the cell and activity starts into being pathogenic bacterial cells.
• As a result of spores heat resistant nature, the sterilization cannot be completed by boiling.
• Hence autoclaving is needed.
• Spores are often not seen in clinical specimens because supply of nutrients is inadequate.
FUNCTION OF BACTERIA
1) FUNCTIONS OF CELL WALL
1. Provides shape to the bacterium
2. Gives rigidity to the organism
3. Protects from environment
4. Provides staining characteristics to the bacterium
5. Contains receptor sites for phages/complements
6. Site of action of antibody and colicin
7. Contains toxic components to host
2)FUNCTION OF CELL MEMBRANE
• 1. Regulates the transport of nutrients and waste products into and
• out of the cell.
• 2. Synthesis of cell wall components
• 3. Assists DNA replication
• 4. Secrets proteins
• 5. Carries on electron transport system
• 6. Captures energy in the form of ATP
STRUCTURE & FUNCTION OF
BACTERIA
 Bacterial Shapes, Arrangements &
Sizes
• Bacteria are classified by shape into three basic groups:
• Based on shape, arrangement and size.
• Shape :
• Cocci (round)
• Bacilli (rod)
• Spirochets (spiral shape)
•Pleomorphic (variable shape)
• Shape is determined by rigid cell wall.
• Microscopy remains the main stay in identification.
Arrangements:
• Diplococci are in short chains (streptococci).
• Grape like clusters (staphylococci).
• Arrangements depend upon orientation and degree of attachment of the
bacteria at the time of cell division.

Size:
• Bacteria range in size from mycoplasma, the smallest bacteria (0.2um), to
bacillus anthracis, one of the largest bacteria.
• Viruses range from polio virus (the smallest virus) to pox virus (the largest virus).
• Yeast are larger than bacteria.
• Size of bacteria varies from 0.2 to 5um.
• Largest bacterial rods are of the same size as of yeast