DISEASE CAUSING AGENTS

• The agents of human infectious diseases belong to five major groups of organisms:
• Bacteria -------------------- Kingdom Prokaryotes
• Fungi -------------------- Kingdom Fungi
• Protozoa -------------------- Kingdom Protista ------------------- Parasites
• Helminths -------------------- Kingdom Animalia ------------------- Parasites
• Viruses --------------------- Acellular
 IMPORTANT FEATURES OF MICROBES
• Structure:
• Cells have a nucleus or nucleoid, which contains DNA; this is surrounded by cytoplasm,
within which proteins are synthesized and energy is generated.
• Viruses have an inner core of genetic material (either DNA or RNA) but no cytoplasm.
• They depend on host cells to provide the machinery for protein synthesis and energy
generation.

• Nature of the nucleic acid:

• Cells contain both DNA and RNA, whereas viruses contain either DNA or RNA but not both.
• Method of replication:
• Cells replicate either by binary fission or by mitosis, during which one parent cell
divides to make two progeny cells while retaining its cellular structure.
• Prokaryotic cells (e.g., bacteria) replicate by binary fission.
• Eukaryotic cells replicate by mitosis.
• Viruses disassemble, produce many copies of their nucleic acid and protein, and
then reassemble into multiple progeny viruses.
• Viruses must replicate within host cells because, they lack protein-synthesizing and
energy-generating systems.
 SHAPE AND SIZE OF BACTERIA
• Bacteria are classified by shape into four basic groups:
• Cocci ---- Round Shaped
• Bacilli ---- Rod Shaped
• Spirochetes ---- Spiral Shaped
• Pleomorphic ---- Variable Shaped

• The shape of a bacterium is determined by its rigid cell wall.

• The microscopic appearance of a bacterium is one of the most important criteria
used in its identification.
• The arrangement of bacteria is important. For example:
• Occur in pairs (diplo-)
• Occur in chains (strepto-)
• Occur in grapelike clusters (staphylo-)
• These arrangements are determined by the orientation and degree of attachment of the
bacteria at the time of cell division.
• Bacteria range in size from about 0.2 to 5 μm.
• The smallest bacteria (Mycoplasma) are about the same size as the largest viruses
(poxviruses) and are the smallest organisms capable of existing outside a host.
• The longest bacteria rods (Thiomargarita magnifica) are the size of some yeasts and human
red blood cells (7 μm).
STRUCTURES IN CYTOPLASM
 CYTOPLASM

• The cytoplasm has two distinct areas when seen in the electron microscope:
• An amorphous matrix that contains ribosomes, nutrient granules, metabolites, and
plasmids.
• An inner, nucleoid region composed of DNA.
 RIBOSOMES
• Site of protein synthesis

• Composed from ribonucleic acid (rRNA) and Proteins -------- Ribonucleoprotein particle.

• Differ from eukaryotic ribosomes in size and chemical composition.

• 70S in size, with 50S and 30S subunits

• The differences in both the ribosomal RNAs and proteins constitute the basis of the
selective action of several antibiotics that inhibit bacterial, but not human protein
synthesis.
 GRANULES

• Contains several different types of granules

• Serve as storage areas for nutrients

• Stain characteristically with certain dyes

• For example, volutin is a reserve of high energy stored in the form of polymerized
metaphosphate (PO3-).

• “Metachromatic” granule (Store metaphosphate).

 NUCLEOID

• Area of the cytoplasm in which DNA is located.

• The DNA of prokaryotes is a single, circular molecule.

• Contains about 2000 genes (Human DNA has approximately 1,00,000 genes)

• No nucleolus, no mitotic spindle, and no histones.

• One major difference between bacterial DNA and eukaryotic DNA is that bacterial
DNA has no introns, whereas eukaryotic DNA does.
 PLASMID

• Extrachromosomal, double-stranded, circular DNA molecules that can replicate

independently of the bacterial chromosome.

• They can be integrated into the bacterial chromosome.

• Occur in both gram-positive and gram-negative bacteria.

• Two types of plasmids:

• Transmissible: Can be transferred from cell to cell by conjugation

• Non-transmissible: Can not be transferred from cell to cell by conjugation

• Plasmids carry the genes for the following functions and structures of medical
importance:

• Antibiotic resistance, which is mediated by a variety of enzymes.

• Resistance to heavy metals, such as mercury which mediate antiseptic effects

• Resistance to ultraviolet light, which is mediated by DNA repair enzymes.

• Pili (fimbriae), which mediate the adherence of bacteria to epithelial cells.

Transposons (Jumping genes):
• Pieces of DNA that move readily from one site to another either within or between
the DNAs of bacteria, plasmids, and bacteriophages.

• They can code for drug resistant enzymes, toxins, or a variety of metabolic enzymes
and can either cause mutations in the gene into which they insert or alter the
expression of nearby genes.

• Transposons typically have four identifiable domains.

• More than one transposon can be located in the DNA; for example, a plasmid can
contain several transposons carrying drug resistant genes.
STRUCTURES OUTSIDE CELL WALL
 CAPSULE
• Gelatinous layer covering the entire bacterium.

• Composed of polysaccharide.

• Sugar components of the polysaccharide vary from one species of bacteria to another.

• The capsule is important for four reasons:

• Determinant of virulence

• Prevent it from ingesting

• Specific identification

• Play a role in the adherence of bacteria to human tissues

 FLAGELLA
• Long, whiplike appendages that move the bacteria toward nutrients and other
attractants, a process called chemotaxis.

• Composed of many subunits of a flagellin protein

• Need adenosine triphosphate (ATP) for movement

• Only certain bacteria have flagella.

• Many bacilli bacteria have flagella ------ Motile

• Most cocci do not have flagella ---------- Non-motile

• Spirochetes has flagellum like structure ------- Axial filament

• Flagella are medically important for two reasons:

• Some species of motile bacteria (e.g., E. coli and Proteus species) are common
causes of urinary tract infections. Flagella may play a role in pathogenesis by
propelling the bacteria up the urethra into the bladder.

• Some species of bacteria (e.g., Salmonella species) are identified in the clinical
laboratory using specific antibodies against flagellar proteins.
• Depending on the arrangement, flagella can be of the following types:

• Atrichous (No flagellum) e.g., Shigella.

• Monotrichous (single polar flagellum), e.g., Vibrio cholerae.

• Lophotrichous (multiple polar fl agella), e.g., Spirilla.

• Peritrichous (flagella distributed over the entire cell), e.g., Salmonella Typhi, E. coli.

• Amphitrichous (single flagellum at both the ends), e.g., Spirillum minus

• Structure:

• 3–20 micro meter in length and 0.01–0.03 micro meter in diameter.

• The main part of the filament is made up of protein subunits called flagellin arranged
in several helices around a central hollow core.

• The flagellum is attached to the bacterial cell body by a complex structure consisting
of a hook and a basal body.

• The basal body bears a set of rings;

• One pair in Gram positive bacteria

• Two pairs in Gram-negative bacteria

• Through these rings bacteria rotates in either clockwise or anticlockwise direction.

• Above the base of filament is the hook, a short curved structure between the external
filament and basal body.

• This part produces a propeller-like repulsion from the revolving flagellum.

• Spirochetes are motile bacteria but without any external flagella. They are motile
due to the presence of an axial filament.

• Axial filament consists of a bundle of flagellum-like structures that lie between the
cell surface and an outer sheath, and connects one end of the cell to the other. They
are sometimes called the endo-flagellates.
• Functions:

• They are primarily responsible for motility of bacteria by chemotaxis.

• They may play a role in bacterial survival and pathogenesis.

• They are highly antigenic, they possess H antigens, and some of the immune
responses to infection are directed against these proteins.

• The flagella of different bacteria differ antigenically. Flagellar antibodies are not
protective but help in serodiagnosis.
 PILLI (FIMBRIAE)
• Hair like filaments that extend from the cell surface.

• Shorter and straighter than flagella

• Composed of subunits of pilin

• Found mainly on gram-negative organisms.

• Pili have two important roles:

• They mediate the attachment of bacteria to specific receptors on the human cell

• Sex pilus, forms attachment between the male (donor) and the female
(recipient) bacteria during conjugation.
• Structure:

• The pili are shorter and straighter than flagella, although the basic structure is same.

• Like flagella, it consists of helics of protein called pilins, arranged around a hollow core but without a motor.

• They are 0.5 micro meter long and 10 nm thick.

• They are antigenic in nature.

• They are classified into four types as follows:

• Type 1: These occur in E. coli, Klebsiella, Shigella, and Salmonella. They are mannose sensitive.
• Type 2: These are present in S. Gallinarum and S. Pullorum, devoid of any hemagglutinating or adhesive
properties.
• Type 3: These are present in some strains of Klebsiella, Serratia, etc. They agglutinate RBC only after
heating and are mannose resistant.
• Type 4: These are mannose resistant and occur in Proteus.
• Sex pili:

• A specialized kind of pili called sex pili is responsible for the attachment of donor and recipient
cells in bacterial conjugation.

• These pili are longer (10–20 micrometer) and vary 1–4 in number.

• The sex pili are of two types:

• F pili:

• They specifically adsorb male specific RNA and DNA bacteriophages.

• They are encoded by sex factor F and fertility inhibition–positive resistance factors (fi-R
factors).

• I pili:

• They adsorb male specific filamentous DNA phages, encoded by col factor and fi-R factor.
• Functions:

• Pili play a major role in the adherence of symbiotic and pathogenic bacteria to
host cells, which is a necessary step in initiation of infection.

• Transfer of bacterial DNA takes place through sex pili during the process of
conjugation.
 BACTERIAL SPORES
• Highly resistant structures formed in response to adverse conditions

• Found in two genera of gram-positive rods.

• genus Bacillus, which includes the agent of anthrax

• genus Clostridium, which includes the agents of tetanus and botulism

• Spore formation (sporulation) occurs when nutrients are depleted.

• The spore forms inside the cell and contains bacterial DNA, a small amount of
cytoplasm, cell membrane, peptidoglycan, very little water, and most importantly, a
thick, keratin like coat that is responsible for the remarkable resistance of the spore
to heat, dehydration, radiation, and chemicals.
 STRUCTURE AND FUNCTIONS OF BACTERIAL CELL ENVELOPE
• The outer layer or cell envelope provides a structural and physiological barrier between the
protoplasm (inside) of the cell and the external environment.
• The cell envelope protects bacteria against osmotic lysis and gives bacteria rigidity and shape.
• The cell envelope primarily consists of two components:
• Cell wall
• Cytoplasmic or plasma membrane
• It encloses the protoplasm, which consists of:
• Cytoplasm
• Cytoplasmic inclusions (mesosomes, ribosomes, inclusion granules, vacuoles)
• A single circular DNA
• Prokaryotic cells almost always are bounded by a fairly rigid and chemically complex
structure present between the cell membrane and capsule/slime layer called the cell wall.
• Peptidoglycan is the main component of the cell wall and is responsible for the shape and
strength of the cell.
• It is a disaccharide and contains two sugar derivatives:
• N-acetylglucosamine
• N-acetylmuramic acid
• These together by short peptide chains.
• N-acetylmuramic acid carries a tetrapeptide side chain consisting of D- and L-amino acids (D-
glutamic acid and L-alanine) with mesodiaminopimelic acid (Gram-negative bacteria) or L-
lysine (Gram-positive bacteria).
• Tetrapeptide side chains are interconnected by pentaglycine bridges.
• Most Gram-negative cell walls lack interpeptide bridge.
• Cell wall provides shape to the cell and protects bacteria from changes in osmotic pressure,
which within the bacteria cell measures 5–20 atmospheres.
• Bacterial cells can be classified into:
• Gram-positive
• Gram-negative
• The cell walls of the Gram-positive bacteria have simpler chemical structures compared to
Gram-negative bacteria.
 A. GRAM POSITIVE CELL WALL
• The Gram-positive cell wall is thick (15–80 nm) and more homogenous than that of the thin (2
nm) Gram-negative cell wall.
• The Gram-positive cell wall contains large amount of peptidoglycan present in several layers
that constitutes about 40–80% of dry weight of the cell wall.
• The Gram-positive cell wall consists primarily of teichoic and teichuronic acids.
• These two components account for up to 50% of the dry weight of the wall and 10% of the
dry weight of the total cell.
• Teichoic acids
• Teichuronic acid
• Teichoic acids:
• Teichoic acids are polymers of poly ribitol phosphate or polyglycerol phosphate
containing ribitol and glycerol.
• These polymers may have sugar or amino acid substitutes, either as side chain or
within the chain of polymer.
• Teichoic acids are of two types:
• Wall teichoic acid (WTA)
• Lipoteichoic acids (LTA)
• They are connected to the peptidoglycan by a covalent bond with the six hydroxyl of
N-acetylmuramic acid in the WTA and to plasma membrane lipids in LTA.
• Teichuronic acid:
• Teichuronic acid consists of repeat units of sugar acids (such as N-acetylmannuronic
or D-glucuronic acid).
• They are synthesized in place of teichoic acids when phosphate supply to the cell
is limited.
• Gram-positive cell wall also contains neutral sugars (such as mannose, arabinose,
rhamnose, and glucosamine) and acidic sugars (such as glucuronic acid and
mannuronic acid), which occur as subunits of polysaccharides in the cell wall.
 B. GRAM NEGATIVE CELL WALL
• The Gram-negative cell wall is much more complex than the Gram-positive cell wall.
• Peptidoglycan content in the Gram-negative cell wall is significantly less than the
Gram positive cell wall.
• Only 1–2 layers of peptidoglycan (2–8 nm) are present just outside the cell
membrane.
• The Gram-negative cell wall outside the peptidoglycan layer contains three main
components:
• Lipoprotein layer
• Outer membrane
• Lipopolysaccharides.
• 1. Lipoprotein layer:
• The lipoprotein layer is mainly composed of Braun’s lipoprotein.
• Braun’s lipoprotein is a small lipoprotein that is covalently joined to the underlying
peptidoglycan and embedded in the outer membrane by its hydrophobic end.
• The lipoprotein stabilizes the outer membrane of the Gram negative cell wall.
• 2. Outer membrane:
• The outer membrane is a bilayered structure.
• Its inner part resembles in composition with that of the cell membrane, while its
outer part contains a distinctive component called lipopolysaccharide.
• The outer membrane and plasma membrane appear to be in direct contact at many
sites in the Gram-negative wall.
• The outer membrane has a variety of proteins as follows:
• Porins
• Outer membrane proteins (OMPs)
• A. Porins:
• The outer membrane has special channels consisting of protein molecules called porins.
• These porins have many functions:
• They permit the passive diffusion of low-molecular weight hydrophilic compounds,
such as sugars, amino acids, and certain ions.
• They exclude hydrophobic molecules
• They serve to protect the cell.
• B. Outer membrane proteins (OMPs):
• These include the following:
• They are responsible for most of the transmembrane diffusion of maltose and
maltodextrins.
• The outer membrane also contains proteins that are involved in the transport of
specific molecules, such as vitamin B12 and iron-siderophore complexes.
• It also contains a limited number of minor proteins, such as enzymes,
phospholipases, and proteases
• C. Lipopolysaccharides:
• Lipopolysaccharides (LPS) are complex molecules present in the outer membrane
of the Gram-negative bacteria.
• Structurally, the LPS consist of three main components:
• Lipid A
• Core oligosaccharide
• O polysaccharide or O-antigen.
• Periplasmic space:
• Periplasmic space is a distinct space between cell membrane and outer membrane
(innermost layer of Gram-negative cell wall) in Gram-negative bacteria.
• This space is filled with a loose layer of peptidoglycan matrix.
• The periplasmic space of Gram negative bacteria contains many proteins that
participate in nutrient acquisition, and many hydrolytic enzymes, beta lactamases
binding proteins, and enzymes that participate in the peptidoglycan synthesis.
• Polymers of D-glucose, called membrane-derived oligosaccharides, appear to play a
role in osmoregulation.
• The periplasmic space is less distinct in Gram positive cell walls.