MICROBIOLOGY LECTURE 1 – Bacterial Morphology and growth and reproduction

Ultrastructure – Dra. Garduque 3. Obligate intracellular parasites- chlamydia and rickettsia

Notes from Lecture 4. Differs from eukaryotes
USTMED ’07 Sec C – AsM a. do not contain 80 s ribosomes or membrane bound
organelles (nucleus, mitochondria, lysosomes,
endoplasmic reticulum or Golgi bodies)
Only two types of cells are produced by all living organisms on b. they lack the 9+2 fibril flagellum or cilia structures
earth. characteristic of eukaryotic cells
1. Prokaryotes (pro. or primitive nucleus) do not have a 5. Have 70 s ribosomes and a naked single circular chromosomes
membrane bound nucleus (nucleiod) composed of DS DNA that replicates amitotically
a. eubacteria (true bacteria) 6. Cytoplasmic membrane of eubacteria - contains ester-
b. archaebacteria (ancient bacteria) linked lipids and carries out transport, energy production
2. Eukaryotes (eu, or true nucleus) have a membrane specialized biosynthetic functions
bound nucleus 7. Motility, if present, is conferred by single filament
flagellar structures
a. Algae
8. Some produce external microfibrils(pili or fimbriae) that
b. fungi
serve adhesive functions
c. protozoa
9. Mycoplasmas do not possess cell walls, whereas other
d. plants
eubacteria produce envelope that contains a chemically
e. animals
similar cell wall peptidoglycan
10. Cell-wall producing eubacteria and archaebacteria may
PROKARYOTES EUKARYOTES
occur as spheres(cocci), rods(bacilli) or curved or spiral-
Size of cell typically 0.20 -2 um in typically 10-100 um in shaped cells
diameter diameter 11. Chemical nature of envelope components and structures-
imparts useful staining characteristics that classifies
usually < 5 um in usually >10 um in eubacteria into: gram positive, gram negative and acid
diameter diameter fast organisms
Nucleus a single DNA molecule true nucleus within a 12. Archaebacteria may differ from the eubacteria in their:
not bounded by a membrane a. chromosome components
membrane b. presence of ether linked membrane lipids
c. absence of eubacterial peptidoglycan
DNA chains replicate divide by mitosis d. possession of different types of metabolism and
and separate producing cofactors
two nuclear bodies

single chromosome, always more than one THE BACTERIAL CELL

circular, naked chromosome
1. Most bacteria produce a layered cell envelope that includes
DNA not complexed DNA complexed with the plasma membrane, cell wall and associated proteins and
with histones histones polysaccharides.
Cytoplasm no cytoskeleton or cytoskeleton; 2. Some bacteria produce capsules or slimes.
cytoplasmic streaming cytoplasmic streaming 3. External filamentous Appendages(flagella and pili) may also
Cytoplasmic structures occur.
Endoplasmic Absent present 4. The cell wall is a rigid structure that encloses and protects
reticulum the protoplast from physical damage and conditions of low
Golgi Absent Present external osmotic pressure.
apparatus 5. The cell wall also generally allows bacteria to tolerate a
Mitochondria Absent present wide range of environmental conditions.
Lysosomes Absent present 6. The protoplast comprises the naked cytoplasmic membrane
Ribosomes 70 s free in the 80 s in the endoplasmic and its contents.
cytoplasm reticulum and free in 7. Internally, bacteria are relatively simple cells.
the cytoplasm; 70 s in 8. Major cytoplasmic structures include a central fibrillar
organelles chromatin network surrounded by an amorphous cytoplasm
Plasma sterols usually absent contains sterols; that contains ribosomes.
membrane except in mycoplasma; CHO serve as receptors 9. Cytoplasmic inclusion bodies or energy storage granules vary
no CHO in chemical nature according to species and in amounts
Cell wall chemically complex; chemically simple; depending on growth phase and environment.
mucopeptides if present, consists 10. Some cytoplasmic structures, such as endospores, are
containing muramic of simple polysacc- limited to only a few bacteria.
acid and DAP or lysine rides or inorganic 11. Typical gram positive and gram negative bacterial cells
substances(cellulose differ primarily in cell envelope organization.
and chitin)
Size of bacteria
Capsule Frequently present Absent
- Unit of microbial measurement
Double Absent Mitochondria,
membrane chloroplasts
organelles 1. micrometers (um)
Simple chlorobium; vesicles Golgi membranes; o 1 um being 10-6 m or 0.000001 m(1/25,000
membrane organelles lysosomes; endoplasmic inch)
organelles chromatophores, reticulum; 2. nanometers
thylakoids vacuoles o 1 nm being 10-9 or 0.000000001 m.
Locomotion flagella that rotate; flagella and cilia that
gliding motion undulate; ameboid - Pathogenic bacterial species vary from approximately
motion 0.4 to 2 um in size
Flagella submicroscopic; microscopic, contain 20
each equivalent to a fibrils (nine pairs
single eukaryotic fibril surrounding two single
fibrils)

consists of two protein complex; consists of

building blocks multiple microtubules

GENERAL PROPERTIES OF BACTERIA

1. Single celled organisms that reproduce by simple division, ie,

binary fission
2. Most are free living and contain the genetic information and
energy producing and biosynthetic systems necessary for
 1. Plasma membrane or cytoplasmic membrane
2. Cytoplasm
3. Nuclear area
4. Ribosomes
5. Inclusions
a. metachromatic granules or Babes Ernst
granules
b. polysaccharide granules
c. lipid inclusions
d. sulfur granules
e. carboxysomes
f. gas vacuoles
6. Endospores
7. mesosomes

Glycocalyx
- a viscous(sticky), gelatinous polymer that is external to
the cell wall
- composed of polysaccharide and polypeptide or both
- chemical composition varies widely with the specie
- made inside the cell and excreted to the cell surface
1. Slime layer - if the substance is unorganized and only
loosely attached to the cell wall
2. Capsule
o if the substance is organized and is firmly
attached to the cell wall
o found in both gram positive and Gram
negative cells
o presence denotes virulence
o encapsulated cells form smooth or mucoid
colonies
o nonencapsulated cells form rough colonies
o not seen in the usual stained smear because of
their failure to retain the dye
o capsular substance is antigenic functioning
largely as a partial antigen or hapten

A. Two simple methods to distinguish the capsule

1. India ink technique - most satisfactory method of
demonstrating the capsule
- Bacteria is suspended in diluted India ink.
- Bacterial cells appear to lie in a lacunae.
Chemical Composition of Bacteria
1. Water - 70% 2. Quellung reaction - Homologous antibody is added to a
2. Dry weight - 30% composed of: preparation of capsule.
a. DNA - 5% MW 2,000,000,000
b. RNA - 12% • microprecipitation at the periphery of the capsule
c. protein- 70% found in: altering its refractive index rendering the capsule to be
i. Ribosomes(10,000) – RNA visible
ii. Protein particles - MW 3,000,000
iii. Enzymes
iv. Surface structures
d. polysaccharides - 5%
e. lipids - 6%
f. phospholipids - 4%

Components of the Bacterial Cell

A. Structures external to the cell wall
1. Glycocalyx
a. Capsules B. Functions of the Capsule
b. slime layer 1. Protection
2. Flagella 2. Identification
3. Axial filaments 3. Vaccine preparation
4. Pili or fimbriae 4. Tissue attachment
5. Antibiotic barrier
B. Cell Wall
C. Structures internal to the cell wall C. Medical Importance - rapid serological identification of:
 1. Several groups of streptococci
2. Meningococcus
3. Hemophilus influenzae
4. Klebsiella pneumoniae
5. Some of the coliforms
6. Yersinia and Bacillus specie
Amphitrichous flagella Peritrichous flagella
Flagella
- organ of locomotion
- long filamentous appendage originating in a spherical
body or basal granules
- occur most commonly although not exclusively among
the rod-shaped bacteria

A. Three morphological regions

1. Helical filament
o long outermost region; composes up to 90% of
its length
o contains the globular (roughly spherical)
protein flagellin arranged in several chains
that intertwine and form a helix around a
hollow core
2. Hooked or curved area
o filament is attached; consists of a different
protein
3. Basal body
o terminal portion of the flagellum
o anchors the flagellum to the cell wall and
plasma membrane
o composed of a central rod inserted into a
series of rings

a. Gram negative - 2 pairs of rings

i. Outer pair - anchored to the outer
membrane (Lring) and peptidoglycan
layer (P ring)
ii. Inner pair - anchored to the plasma
membrane (SM ring)
b. Gram positive - only inner pair is present

B. Types of arrangement of flagella

1. Atrichous - no flagella C. Motility
2. Monotrichous - one polar flagellum - presumptive evidence that bacteria possesses flagella
3. Lophotrichous - tuft of several polar flagella although it gives no indication of the number or
4. Amphitrichous - flagella at both poles rearrangement of the flagella
5. Peritrichous - flagella over the entire cell surface
1. Types of bacterial motility
a. run or swim - when a bacterium moves in one
direction for a length of time
b. tumbles - periodic, abrupt random changes in
direction
c. swarming - rapid wavelike growth across a solid
culture medium
2. Two ways by which motility can be demonstrated
a. direct or microscopic
o Hanging drop preparation or wet mount
preparation
o Distinguishes:
i.Brownian movement - when the
bacteria show molecular movement
with no apparent effort to change
their position and direction to the
field
ii. true motility - if a bacterium
describes a rotatory, undulatory or
sinuous movement
b. indirect or macroscopic
o Stab inoculation of the semisolid media
i. nonmotile - growth is limited at the
point of inoculation
ii. motile - growth is diffuse or moves
away from the line of inoculation;
turbidity of the medium

c. Mechanism of flagellar movement - relative

rotation of the rings in the basal body of the
flagellum

D. Antigenicity
- flagellar or H antigen - useful in the serological
identification of serotypes of Salmonella organisms

Axial filaments
Monotrichous flagella Lophotrichus flagella - bundles of fibrils that arise at the ends of the cell
beneath the outer sheath and spiral around the cell
 - rotation moves an opposing movement of the outer a. teichoic acid - polymeric complexes of ribitol
sheath that propels the spirochetes by causing them to phosphate or glycerol phosphate
move like corkscrews o Functions:
i. Binds and regulate the movement of cations
(positive ions) into and out of the cell
ii. Assumes a role in cell growth
iii. Storage of phosphorus
iv. Regulates the activity of autolysins thereby
Found in Spirochetes and are preventing extensive wall breakdown and
similar to flagella, but are possible cell lysis (rupturing)
located between the cell wall v. Provides much of the wall’s antigenic
and an outer sheath, and are specificity making it possible to identify
attached to one end of the bacteria by serological means
organism.
b. Polysaccharides
o Streptococci - C- CHO - immunologically active
Fimbriae and pili substance present in the cell wall; responsible
- hairlike appendages that are shorter, straighter and for their serological differences and the basis
thinner than flagella of Lancefield Classification (A to U)
- used for attachment rather than for motility
- consist of a protein called pilin arranged helically c. proteins
around a central core o not an internal component of the cell wall but
maybe in a microcapsular or capsular layer
A. Fimbriae o Ex. M proteins of streptococci glutamyl
- can occur at the poles of the bacterial cell or they can polypeptides of Bacillus specie
be evenly distributed over the entire surface of the cell
- allow a cell to adhere to surfaces

B. Pili
- usually longer than fimbriae and number only one or two
per cell
- genetically determined by a fertility factor called F
factor which is carried on an episome (sex or F pilus)

- Two Types:

1. Sex or conjugation pili for the

transfer of extrachromosomal DNA
between donor and recipient

2. Attachment pili or fimbriae.

There are many and are used
for attachment to surfaces.
- Functions of Pili:
1. Sites of adsorption for RNA and DNA viruses
2. Act as a means of genetic transfer between similar or
related Gram negative enteric species
3. Provide the channel through which DNA from the donor
(male) cell is transferred to the recipient (female cell)
Cell wall
- major structural component of all microorganisms
- responsible for the shape of the microorganism
- a strong and rigid structure that protects and supports
the weaker and biochemically more active parts of the
cell
- high tensile strength is attributed to a layer composed
of a substance variously referred to as murein,
mucopeptide or peptidoglycan layer
A. Composition
1. Peptidoglycan layer
- component in both gram positive and gram negative
cells that provides the structural support for the cell
wall
- Components of the peptidoglycan layer:
a. backbone of polysaccharide - consists of
alternating residues of amino sugars, N-
acetylglucosamine and N-acetylmuramic acid
held by beta 1,4 linkages
b. a set of identical tetrapeptide side chains
attached to N- acetylmuramic acid
c. a set of identical peptide cross bridges
2. The gram positive cell wall
- thickness of the cell wall varies from 150 to 500 A
- many layers of peptidoglycan forming a thick rigid
structure
- peptidoglycan layer - 50 to 90% of the wall
- rest of the wall is made up of:
3. The Gram negative cell wall
- more complex than that of Gram positive cell wall
- peptidoglycan layer makes up only a portion of the wall
- 5 to 10%
- one or very few layers of peptidoglycan layer and an
outer membrane
- peptidoglycan layer is bounded to the lipoproteins in the
outer membrane
- peptidoglycan is embedded in a soft material, the
periplasmic gel, which contains a high concentration of
degradative enzymes and transport proteins
- does not contain teichoic acid
- more susceptible to mechanical breakage due to a small
amount of peptidoglycan layer
- outer membrane consists of:
a. lipoproteins
o Composed of protein covalently linked to lipid
o Firmly bind the outer membrane to the
underlying peptidoglycan layer via covalently
bonds
b. Lipopolysaccharides
i. Polysaccharide portion of LPS - composed of
sugars called O polysaccharides- functions as
antigens and are useful for determining
species of gram negative bacteria
ii. Lipid portion of LPS - Lipid A - referred to as
endotoxin and is toxic when in the host’s
bloodstream or gastrointestinal tract
c. phospholipids
  difficult to cultivate
 produced more readily by penicillin
than with lysozyme
 capable of reverting to the normal
bacillary form upon the removal of
the inducing stimulus
 infections are resistant to antibiotic
treatment since they become
sequestered in the protective
regions
2. interference in the formation of peptide cross
bridges of peptidoglycan thus preventing the
formation of a functional cell wall
Ex. Penicillin
- Functions of the outer membrane
Plasma (Cytoplasmic membrane or inner membrane)
1. Effective hydrophobic barrier to the passage of a large
number of molecules including fatty acids, lysozymes,
- a thin structure lying inside the cell wall and enclosing
the cytoplasm of the cell
detergents and in particular antibiotics
2. Blocks the escape of hydrolytic enzymes which
- consists primarily of phospholipids which are the most
abundant in the membrane and proteins
accumulate between the peptidoglycan layer and the
A. Structure
outer membrane
3. An intact outer membrane carrying the O antigens helps - Electron microscopy - two layered structure;
prevent the interaction of complement and antibiotics phospholipid bilayer; phospholipid molecules arranged in
on its surface two parallel rows
4. Makes bacteria vulnerable to attack by providing 1. Phospholipid molecule
attachment sites for viruses and bacteriocins a. polar head - composed of a phosphate group
5. Permeability due to porins - form channels; permit the and glycerol that is hydrophilic(water loving)
and soluble in water
passage of molecules such as nucleotides, peptides,
amino acids, vitamin B12 and iron b. nonpolar tail - composed of fatty acids that
6. Its strong negative charge is an important factor in are hydrophobic (water fearing) and are
insoluble in water; lies in the interior of the
evading phagocytosis and the action of complement,
bilayer
two components of the defenses of the host.
2. Protein molecule
a. peripheral proteins
B. Functions of the cell wall
o easily removed from the membrane by mild
1. Maintains the cell’s characteristic shape - the rigid wall
treatment
compensates for the flexibility of the phospholipids
membrane and keeps the cell from assuming a spherical o lie at the inner or outer surface of the
shape membrane
2. Countering the effects of osmotic pressure – the o Functions:
strength of the wall is responsible for keeping the cell i. Act as enzymes that catalyze chemical
from bursting when the intracellular osmolarity is much reactions
greater than the extracellular teichoic osmolarity ii. Act as a scaffold for support
iii. Act as mediators of changes in
3. Provides attachment sites for bacteriophages – teichoic membrane shape during movement
acids attached to the outer surface of the wall are like
b. integral proteins
landing pads for viruses that infect bacteria
4. Provides a rigid platform for surface appendages  can be removed from the membrane
-flagella, fimbriae, and pili all emanate from the wall only after disrupting the bilayer
and extend beyond it  penetrate the membrane completely
 contain channels through which
C. Atypical cell walls
substances enter and exit the cell
- naturally occurring wall less microorganisms
1. Genus Mycoplasma and related organism
o smallest known bacteria that can grow and
reproduce outside living host cells
o pass through most bacterial filters and were
first mistaken for viruses
o require cholesterol or sterols for growth
o cultivated in the laboratory only in hypertonic
media
2. Archaebacteria
o lack walls or may have unusual walls
composed of polysaccharides and proteins but
not peptidoglycan
o walls contain N-acetylalosaminuronic acid but
lacks the D amino acids found in the bacterial
cell walls

D. Damage to the cell wall

1. Hydrolysis with lysozyme
a. Protoplast
 Gram positive cell to a wall less
form
 no cell wall material is retained
 spherical and is capable of carrying
on metabolism
b. spheroplast
 Gram negative cell to a wall less
form
 some cell wall materials are
retained
c. L forms
 tiny mutant bacteria with defective
cell walls
 2. Ribosomes
3. Inclusions

Nuclear area or nucleoid

A. Bacterial chromosomes - single long, circular molecules of
double stranded DNA
1. Carries all the information required for the cell
structures and functions
2. Bacterial chromosomes do not include histones and are
not surrounded by a nuclear envelope (membrane)
3. Attached to plasma membrane, the proteins of which
are responsible for the replication of DNA and
segregation of the new chromosomes to daughter cells
in cell division
B. Plasmids - small, circular, double stranded DNA molecules
1. Extrachromosomal genetic elements
2. Replicate independently of chromosomal DNA
3. Associated with plasma membranes
4. Usually contain from five to 100 genes
5. Maybe gained or lost without harming the cell
6. May carry genes for such activities as:
a. antibiotic resistance
b. tolerance to toxic metals
c. production of toxins
d. synthesis of enzymes
7. Can be transferred from one bacterium to another

Ribosomes
- sites for protein synthesis
- composed of two subunits, each subunit being composed
of protein and a type of RNA called ribosomal RNA
(rRNA)
- prokaryotic ribosomes are called 70s ribosomes
- MA: inhibition of protein synthesis on the ribosomes Ex.
Streptomycin, tetracyclines

Inclusions
- reserve deposits

A. metachromatic granules or volutin or Babes Ernst granules

- spherical granules that are not membrane bound
B. Functions of the plasma membrane - contains relatively large amounts of trichloroacetic acid-
1. Holds the intracellular contents within the cytoplasm insoluble polymetaphosphate or polymerized phosphoric
and prevents their leakage acid or polymerized polymetaphosphate or volutin
2. Concentrates nutrients by effecting their transport from - generally formed by cells that grow in phosphate rich
the external environment of the cell to the cytoplasm of environments
the cell - found in algae, fungi and protozoans as well as bacteria
3. Provides the enzymes necessary for capsules, cell wall - stain red with certain blue dyes such as methylene blue
and cell membrane synthesis - characteristic of Corynebacterium diptheriae
4. Serves as the site for enzymes involved in electron B. Polysaccharide granules- not usually enclosed by a membrane
transport and energy metabolism - consist of glycogen and starch
- glycogen granules - reddish brown with iodine
C. Associated structures - starch granules - blue with iodine
1. Chromatophores or thylakoids C. Lipid inclusions
o infoldings of the plasma membrane that - appear in various species of Mycobacterium, Bacillus,
extend into the cytoplasm Azobacter, Spirillum
o contains the pigments and enzymes involved - polymer polybetahydroxybutyric acid - storage material
in photosynthesis D. sulfur granules
2. Mesosomes - sulfur bacteria – (Genus Thiobacillus) deposit sulfur
- one or more large, irregular folds in the plasma granules in the cell, where they serve as an energy
membrane; believed to be artifacts reserve
- Functions of Mesosomes: E. Carboxysomes
a. site for energy metabolism and respiration
- polyhedral and hexagonal inclusions
b. site for attachment of DNA to membrane and
are the site of growing septum formation - contain the enzyme ribulose 1,5 diphosphate
carboxylase
c. contain enzymes instrumental in the synthesis
of the cell wall, cell membrane and other - required by bacteria for carbon dioxide fixation during
components external to the cytoplasmic photosynthesis
membrane - Ex. Nitrifying bacteria, cyanobacteria, thiobacilli
d. site of exoenyzme synthesis and secretion for F. Gas vacuoles
penicillinase - hollow cavities found in many aquatic prokaryotes
- consist of rows of several individual gas vesicles which
Cytoplasm
are hollow cylinders covered by protein
- the internal matrix of the cell contained inside the - Function: maintain buoyancy so that the cells can
plasma membrane
remain at the depth in the water appropriate for them
- thick, aqueous, semitransparent and elastic
to receive sufficient amounts of oxygen, light and
nutrients
A. Composition
1. Water - 80%
Endospores
2. Proteins (enzymes)
- a refractile oval body formed within the bacterial cell
3. Carbohydrates
found intracellularly and extracellularly in the usual
4. Lipids
stained smear
5. Inorganic ions
- found in all species of Family Bacillaceae
6. Low molecular weight compounds
o Genus Bacillus - aerobic sporeforming rods
B. Major Structures
1. DNA o Genus Clostridium - anaerobic sporeforming
 rods
- found also in Sporosarcinae- Gram positive coccus Cocci Neisseria
Coxiella Burneti Rods
Facultative
A. Structure of the endospores Straight
Respiratory Haemophilus
1. Core or spore protoplast – contains: organisms Bordetella
a. complete nucleus Leigionella
b. all of the components of the protein Zoonoitic Brucella
synthesizing apparatus organisms Francisella
c. energy generating systen on glycolysis- energy Pasteurella
for germination- stored as 3 phosphoglycerate Enteric & Escherichia
2. Spore wall related Serratia
organisms Klebsiella
a. innermost layer surrounding the inner spore Salmonella
membrane Shigella
b. contains the normal peptidoglycan Proteus
c. Becomes the cell wall of the germinating Curved Camphylobacter
vegetative cell Vibrio
3. Cortex Aerobic Pseudomonas
a. thickest layer of the spore envelope with Anaerobic Bacteroides
many fewer cross links than are found in cell
wall peptidoglycan 3. Acid fast -Mycobacteria
b. Cortex peptidoglycan is extremely sensitive to
lysozyme B. Non-free living (Obligate intracellular parasites)
-Rickettsia
c. Contains dipicolinic acid, mucopeptide and -Chlamydia
calcium, all of which are significant in spore
resistance II. Flexible, thin walled cells (Spirochetes)
4. Coat - composed of keratin like portion containing -Treponema
many intramolecular disulfide bonds -Borellia
5. Exosporium - lipoprotein membrane containing some -Leptospira
carbohydrate
III. Wall-less cells -Mycoplasma
B. Medical Importance of Spores

1. Sporeforming pathogens may cause human illness.

a. Clostridium botulinum – botulism
b. Clostridium tetani – tetanus
c. Clostridium perfringens- gas gangrene
d. Bacillus anthracis – anthrax
e. Coxiella burneti - Q fever
2. Sporulation process provides medicine a number of
antibiotics
a. Bacitracins
b. Gramicidins
c. Tyrocidins

Classification of Medically Important bacteria Identification of Bacteria

I. Rigid, thick walled cells 1. Isolation of organisms in pure (axenic) Culture

2. Bacterial Colony Morphology
A. Free living(extracellular) 3. Microscopic Morphology and Staining Reactions
4. Biochemical Characteristics
1. Gram positive 5. Antibiotic susceptibility
6. Epidemiologic Phage and Serotyping
Cocci Streptococcus
Staphylococcus -end-
Sporeforming rods
Aerobic Bacillus ustmedc3@yahoogroups.com
Anaerobic Clostridium audsmartinez@gmail.com
Nonsporeforming rods
Nonfilamentous Corynebacterium
Listeria
Filamentous Actinomyces
Nocardia

2. Gram negative