Characterization and

classification
These are the group of microscopic single
celled organism that live enormous number in
almost every environment on the surface of
earth, from deep vents to the digestive tracts of
humans.
It can be classified and characterized on the
bases of morphology, biochemical
characterization, physiological
characterization……………..
SHAPE………….
Shapes……………
Coccus
Chain = Streptoccus
Cluster = Staphylococcus
Bacillus
Chain = Streptobacillus
Coccobacillus

Vibrio = curved

Spirillum

Spirochete

Square

Star
Flagella, Fimbriae, pili……..
Short protein appendages
 smaller than flagella
Adhere bacteria to surfaces
 E. coli has numerous types
K88, K99, F41, etc.
 Antibodies to will block adherance
F-pilus; used in conjugation
 Exchange of genetic information
Flotation; increase boyancy
 Pellicle (scum on water)
 More oxygen on surface
Gram-positive cell walls Gram-negative cell
walls
 Thin peptidoglycan
 Thick peptidoglycan
 5-10% peptidoglycan
 90% peptidoglycan
 No teichoic acids
 Teichoic acids
 3 layers
 1 layer
 Outer membrane has
 Not many polysaccharides
lipids, polysaccharides
 In acid-fast cells,
 No acid- fast cells
 contains mycolic acid
(mycolic acid)
 Pseudomonas

Neisseria

Streptococcus
Attach to outer surface of cell wall
“Glycocalyx” (gly-koh-kay’licks) is the general term of
the polysaccharide capsule/ slime layer
Produced when proper growth conditions
 Excess of nutrients
Made from a variety of polysaccharides
 Poly D-glutamate in Bacillus sp.
 Capsule: the polysaccharide that has a uniform
thickness & can be thicker than cell
 Streptococcus pneumoniae  infectious to human,
capsule help protect the cell against phagocytosis
 Can be observed by performing Negative staining
with India Ink
 Slime layer: the polysaccharide adhered to the
cell wall in a diffuse arrangement
 Bacteria that cause dental carries  help bacteria to
colonise surface
 Flagella
 Pili
 Capsule
 Plasma Membrane
 Cytoplasm
 Cell Wall
 Lipopolysaccharides
 Teichoic Acids
 Inclusions
 Spores
Colonial Morphology………….
• Shape
• Edge
• Elevation
• Size
• Chromogenesis
• Opacity
• Surface
• Consistency
• Emulsifiability
• Odor
 Carbon Sources
Autotrophs CO2 sole or principal biosynthetic
carbon source.
Heterotrophs Reduced, preformed, organic
molecules from other organisms
 Energy Sources
Phototrophs Light
Chemotrophs Oxidation of organic or inorganic
compounds (chapter 9)
 Electron Sources
Lithotrophs Reduced inorganic molecules
Organotrophs Organic molecules
. Psychrophiles
-- cold loving microbes
2. Mesophiles
--moderate-temperature loving
organism
-- most pathogens and indigenous flora
3. Thermophiles
-- heat loving microbes
-- ex. Thermophilic cyanobacteria
found in hot springs
-- Thermodurics: organisms that can
survive or endure boiling
-- ex. Endospores and viruses
1. Minimum Growth temperature
- lowest temperature at which the species
will grow
2. Optimum Growth temperature
- temperature at which the species grows
best
3. Maximum Growth temperature
- highest temperature at which growth is
possible
Classification Temperature Range (°C) Optimum
Growth Temperature
(°C)

Psychrophile -10 to 20 10
Psychrotroph 5 to 30 25
Mesophile 10 to 45 37
Thermophile 40 to 75 55
Hyperthermophile 65 to 120 90-100
Acidity or alkalinity of a solution
1. Neutrophiles
-- neutral growth medium (pH 7)
-- most microorganisms
2. Acidophiles
--prefer a pH of 2-5
--microbes that can live in the stomach
3. Alkaliphiles (Basophiles)
--prefer pH greater 8.5
-- found in intestine
-Based on relationship to O2
1. Aerobes---use molecular O2 for life and
reproduction
a. Obligate aerobes
- require an atmosphere that contains O2
similar to room air (20-21% O2), Ex. Mycobacteria
b. Microaerophiles
- require O2 lower than room air (=5% O2)
- ex. Neisseria, Campylobacter
2. Anaerobes
- do not require O2 for life and reproduction
- vary based on sensitivity to O2
a. Obligate anaerobe
- unable to grow in O2, ex. Clostridium
 Source of Energy
--Phototrophs---light
--Chemotrophs– inorganic or organic compounds
 Source of Carbon
-- Autotrophs---CO2
-- Litotrophs—inorganic compound except CO2
-- Heterotrophs (Organotrophs) ---Organic compounds
 Energy Source and Carbon Source
--Photoautotrophs---Light + CO2
---ex. Plants, algae, cyanobacteria, purple and green
sulfur bacteria
--Photoheterotrophs (Photoorganotrophs) --- Light + organic
compounds
—ex. Green and purple non-sulfur bacteria
--Chemoautotrophs– Chemical + CO2
ex. Nitrifying, hydrogen, iron and sulfur
bacteria
--Chemolitotrophs--- Chemical + inorganic
compound except CO2
-- Chemoheterotrophs– Chemical + organic
compound
ex. All animals, protozoa, fungi, most
bacteria
-- Photolithotrophs – Light + inorganic compound
except CO2
ex. Plants and algae: producers of food and
O2 for chemoheterotrophs
Bacteria are
susceptible to
certain
antibiotics which
can be tested by
sensitivity test.
Kurby baur
method is used
to test the
susceptibility, by
diffusion
method.
Indole
Methyl Red
Citrate
H2S production
Urea hydrolysis
Motility
Lactose fermentation
Sucrose fermentation
Glucose fermentation & gas production
How to Perform Test: Inoculate Tryptone broth with
inoculating loop.
Property it tests for: This test is performed to help
differentiate species of the family Enterobacteriaceae. It
tests for the bacteria species’ ability to produce indole.
Bacteria use an enzyme, tryptophanase to break down the
amino acid, tryptophan, which makes by-products, of
which, indole is one.
Media and Reagents Used: Tryptone broth contains
tryptophan. Kovac’s reagent—contains hydrochloric acid,
dimethylaminobenzaldehyde, and amyl alcohol—yellow in
color.
Reading Results: Kovac’s reagent reacts with indole
and creates a red color at the top part of the test tube.
How to Perform Tests: Inoculate glucose broths
with inoculating loop. After 48 hours of incubation, add a
few drops of MR to tube.
Properties they test for: tests is used to help
differentiate species of the family Enterobacteriaceae.
 MR—tests for acid end products from glucose fermentation.

Media and Reagents Used:

 Glucose Broth
 Methyl Red indicator for acid
Reading Results:
MR— a + result is red (indicating pH below 6) and a – result is yellow
(indicating no acid production)
 How to Perform Test: Inoculate slant with inoculating loop.
 Property it tests for: This test is used to help differentiate
species of the family Enterobacteriaceae. It is selective for bacteria
that has the ability to consume citrate as its sole source of carbon and
ammonium as sole nitrogen source.
 Media and Reagents Used: Simmon’s Citrate Agar contains
sodium citrate (carbon source), ammonium ion (nitrogen source), &
pH indicator—bromthymol blue.
 Reading Results:
 A + result is blue (meaning the bacteria metabolised citrate and produced
an acid end product) and a – result remains green
Left positive and right negative
How to Perform Test: Stab media with inoculating
needle.
Property it tests for: This test is used to help
differentiate species of the family Enterobacteriaceae. This
test is used to determine the ability to reduce sulfur into
H2S.
Media and Reagents Used: media contains the
sulfur containing amino acid, cysteine, sodium thiosulfate,
& peptonized iron or ferrous sulfate.
Reading Results: H2S will react with the iron or ferrous
sulfate and produce a black precipitate. A positive result
has a black precipitate present and a negative result has no
black precipitate.
 How to Perform Test: Inoculate Urea broth with
 inoculating loop.
 Property it tests for: This test is done to
 determine a bacteria’s ability to hydrolyze urea to
 make ammonia using the enzyme urease.
 Media and Reagents Used: Urea broth
 Contains a yeast extract, monopotassium phosphate,
 disodium phosphate, urea, and phenol red indicator.
 Reading Results: Urea broth is a yellow-orange color. The
enzyme urease will be used to hydrolyze urea to make ammonia. If
ammonia is made, the broth turns a bright pink color, and is positive.
If test is negative, broth has no color change and no ammonia is
made.
How to Perform Test: Stab motility media with
inoculating needle.
Property it tests for: This test is done to help
differentiate species of bacteria that are motile.
Media and Reagents Used: Motility media
contains tryptose, sodium chloride, agar, and a color
indicator.
Reading Results: If bacteria is motile, there will
be growth going out away from the stab line, and test
is positive. If bacteria is not motile, there will only be
growth along the stab line. A colored indicator can be
used to make the results easier to see
 From left to right:
+ – +
 How to Perform Test: Inoculate lactose broth with inoculating
loop.
 Property it tests for: This tests for the bacteria’s ability to
ferment lactose.
 Media and Reagents Used: Lactose broth contains beef
extract, gelatin peptone, and lactose. A phenol red indicator is added
to indicate acid production from fermentation.
 Results
 A positive result is yellow after indicator is added (indicating
lactose fermentation)
 A negative result will have no color change or will be redish
 How to Perform Test: Inoculate sucrose broth with inoculating
loop.
 Property it tests for: This test is done to help differentiate
species of the family Enterobacteriaceae. This tests for the bacteria’s
ability to ferment sucrose and production of acid end-product
 Media and Reagents Used: Sucrose broth contains beef
extract, gelatin peptone, and sucrose. Phenol red indicator is added
to indicate an acid end-product.
 Results
 A positive result is yellow after indicator is added (indicating
sucrose fermentation)
 A negative result has no color change or is reddish.
 How to Perform Test: Inoculate broth with inoculating loop.
 Property it tests for: This test is done to help differnetiate
species of the family Enterobacteriaceae. This tests for the bacteria’s
ability to ferment glucose and produce gas and/or an acid end-
product..
 Media and Reagents Used: Glucose broth contains beef
extract, gelatine peptone, and glucose. A phenol red indicator is
added to indicate an acid enproduct. A Durham tube is added to
indicate gas production.
 Results
 A positive result for acid is yellow after indicator is added
(indicating glucose fermentation)
 A positive result for gas is a bubble in the Durham tube.
 A completely negative result has no color change or reddish color
and no bubble.
Thank you………………….