PART 1

Prokaryotic Microbes
Contains 23 phyla, 32 classes, 5
subclasses, 77 orders, 14 suborders, 182
families, 871 genera, and 5,007 species.
Phenotypic categories

• Gram-negative and have cell wall

• Gram-positive and have cell wall
• Lack cell wall.
 Cell morphology
 Staining reactions
 Motility
 Colony morphology
 Atmospheric requirements
 Nutritional requirements
 Biochemical and metabolic activities
 Specific enzymes that the organism produces
 Pathogenecity
 Genetic composition
Shapes and Arrangements of Bacteria
(Separate Presentation)
Bacteria can lose their characteristic
shape because of adverse growth
condition which prevents the production of
normal cell wall.
Some can revert back to normal shape
while others cannot.
Mycoplasma do not have cell walls.
Pleomorphism- ability to exist in variety of
shapes.
Mycoplasma pneumoniae
Preparation of smear
Heat fixation
Methanol fixation
Purpose of fixation:

• Kills the organism

• Preserves their morphology
• Anchors the smear to the slide
Simple staining
• Determine the morphology
• Dye is applied to the fixed smear, rinse, dried and
examined using OIO.
Structural staining
• Use to observe bacterial capsules, spores, and
flagella.
Differential staining
• Gram staining
• Acid-fast staining
Gram-positive & Gram-negative
 Difference between Gram-positive
and Gram-negative Bacteria

Gram Positive Gram Negative

Color Blue-to-purple Pink-to-red
Peptidoglycan Thick layer Thin layer

Teichoic acids and

lipoteichoic acids in Present Absent
cell walls
Lipopolysaccharide Absent Present
in cell walls
 Clostridium tetani Bacillus subtilis

Gram-positive bacteria tend to be killed by penicillin and detergents.

Gram-negative bacteria are more resistant to antibiotics.
Use to identify Mycobacteria spp.
Carbol fuschin (bright red) is driven into
the bacterial cell using heat.
Heat softens the waxes of the cell walls of
Mycobacteria, enabling the stain to
penetrate.
A decolorizing agent (acid-alcohol) is
then used in an attempt to remove the red
color from the cells.
Because Mycobacteria are not decolorized
by the acid-alcohol, they are said to be
acid-fast.
Most bacteria are non acid-fast.
Acid-fast stain is especially used in TB
labs.
Developed in 1882 by Paul Ehlrich.
Acid-fast staining: Mycobacterium leprae
Negative staining is useful for capsules.
Endospores staining- heat is required to
drive a stain into endospores.
Flagella staining requires a mordant to
make the flagella wide enough to see.
Motileand nonmotile
Presence of flagella, axial filaments or
capable of gliding motility.
Endoflagella
Inspirochetes
Anchored at one
end of a cell
Rotation causes
cell to move
Bacterial colony- mound or pile of bacteria
in a solid culture medium.
Contains million of organisms.
Colony morphology varies from one
species to another.
• Size
• Color
• Over-all shape
• Elevation
• Margin
Oxygen (O2)

Obligate Facultative Obligate Aerotolerant

Microaerophiles
aerobes anaerobes anaerobes anaerobes
CO - Canophiles
2
Nitrogen
• In amino acids, proteins
• Most bacteria decompose proteins
• Some bacteria use NH4+ or NO3−
• A few bacteria use N2 in nitrogen fixation
Sulfur
• In amino acids, thiamine, biotin
• Most bacteria decompose proteins
• Some bacteria use SO42− or H2S
Phosphorus
• In DNA, RNA, ATP, and membranes
• PO43− is a source of phosphorus
Organic Growth Factors
• Organic compounds obtained from the
environment
• Vitamins, amino acids, purines, pyrimidines
Carbon

• Structural organic molecules, energy source

• Chemoheterotrophs use organic carbon sources
• Autotrophs use CO2
Trace Elements
• Inorganic elements required in small amounts
• Usually as enzyme cofactors
Capsules
Pili
Endotoxins
Exotoxins
Exoenzymes
Nuclear area (nucleoid)
Obligate intracellular parasites:
• Ehrlichia and Anaplasma spp. Tick-borne,
ehrlichiosis
• Rickettsia. Arthropod-borne, spotted fevers
R. prowazekii Epidemic typhus
R. typhi Endemic murine typhus
R. rickettsii Rocky Mountain Spotted Fever
Human pathogens:
• Bartonella Trench fever
• B. hensela Cat-scratch disease
• Coxiella Q fever transmitted via aerosols or
milk
 Chlamydia
 Energy parasites
 Mode of transmission:
inhalation of aerosols
or direct contact
between hosts
 C. trachomatis
• Trachoma
• STD, urethritis
 C. pneumoniae
 C. psittaci
• Causes psittacosis
 Mycoplasma
 Wall-less, pleomorphic
 0.1 - 0.24 µm
 M. pneumoniae
 Pleuropneumonia-like
organsims (PPLO)
 Tiny colonies (fried egg
colonies)
 Resistant to antibiotics
Thiomargarita namibiensis- largest
bacteria (750 µm)
Epulopiscium fishelsonii- another
enormous bacteria (80 µm X 600 µm)
Nanobacteria- less than 1 µm, found in
soil, minerals, ocean water, dental plaque,
and even rocks (meteorites)
Thiomargarita namibiensis
Epulopiscium fishelsonii
Nanobacteria
Includespurple bacteria, green bacteria
and cyanobacteria.
Oxygenic photosynthesis
Anoxygenic photosynthesis

light
2H2O + CO2 (CH2O) + H2O + O2

light
2H2S + CO2 (CH2O) + H2O + 2S0
Cyanobacteria
purple sulfur bacteria green sulfur bacteria
Discovered in 1977
Genetic contains 2 phyla, 8 classes, 12
orders, 21 families, 69 genera, and 217
species.
Genetically, archaeans are closely related
to eukaryotes than bacteria.
Thermophiles
 Heat-loving
archaebacteria found
near hydrothermal
vents and hot springs
 Many thermophiles
are chemosynthetic
using dissolved sulfur
or other elements as
their energy source
and iron as a means
of respiration
Halophiles
 Thrive in unusually
salty habitats. Some
can thrive in water
that’s 9% salt; sea
water contains only
0.9% salt.
 Have light-sensitive
pigment
bacteriorhopsodin
which absorbs energy
from sunlight
 Example: Salt Lake
and Dead Sea
Psychrophiles

likeextremely cold
temperatures (even
down to -10
degrees Celsius).
Live in arctic and
antartic oceans
Methanogens

 are anaerobic
archaebacteria that
produce methane
 are found in sewage
treatment plants,
bogs, and the
intestinal tracts of
ruminants.
 ancient methanogens
are the source of
natural gas.
Next topic: Part 2 Eukaryotic
Microbes