Classification and

Identification of Bacteria
 OBJECTIVES
1. Define classification, identification, species, genus, and binomial
nomenclature.

2. Identify the characteristics of microorganisms.

3. Describe how genetic alterations and diversity provide a mechanism for

the evolution and survival of microorganisms.

4. Describe how the classification, naming, and identification of organisms

play a role in diagnostic microbiology in the clinical setting.
SYSTEMS USED TO CLASSIFY BACTERIA:

1. Biological classification
- based on observable characters

a. physiological
b. immunological
c. ecological

Division Protophyta

Class: Schizomycetes
Orders: 1 Pseudomondales
2 Eubacterales
3 Actinomycetales
4 Spirochaetales
5 Mycoplasmaetales
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SYSTEMS USED TO CLASSIFY BACTERIA:

2. Morphological classification:
- based on main characters of each order, families and genera

Two groups
A. Higher bacteria:
- filamentous and grow by branching to form mycelium,
e.g. antinomycetes
- organisms producing true mycelium among actinomycetales
are further classified into:
a. Vegetative mycelium fragments into bacillary or
coccoid element - Gram positive.

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SYSTEMS USED TO CLASSIFY BACTERIA

Types of Vegetative mycelium fragments into bacillary or coccoid

element:
i. Anaerobic, acid fast, e.g. nocardia
ii. Anaerobic non-acid fast,
e.g. Actinomyces israelii; Actinomyces bovis

b. Vegetative mycelium does not fragment into bacillary or coccoid form.

- Conida are formed in chain from aerial hyphae, e.g. streptomyces.

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SYSTEMS USED TO CLASSIFY BACTERIA
B. Lower or true bacteria
- unicellular and never form mycelium.
- grouped on the basis of their shape:
a. Cocci—(from kakkos, meaning
berry) spherical
b. Bacilli—rod shaped
c. Vibrio—Comma shaped
d. Spirilla—Spiral twisted non-
flexous rods
e. Spirochaetes—Thin spirally
twisted, flexous rods
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SYSTEMS USED TO CLASSIFY BACTERIA:

a. Cocci: Follows types of arrangement

Diplococcus: Binary fission occurs in one plane,

e.g. pneumococci

Streptococcus: Cocci are arranged in chain,

e.g. Streptococcus pyogenes, Streptococcus viridans

Staphylococcus: Cocci are arranged in cluster,

e.g. Staphylococcus aureus, Staphylococcus albus

Tetracoccus: Arrangement of cocci in group of four,

e.g. Micrococcus tetragena
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GRAM POSITIVE AND GRAM-NEGATIVE COCCI

COCCI

GRAM NEGATIVE GRAM NEGATIVE

▪ NEISSERIA GONOCOCCUS ▪ STAPHYLOCOCCUS
▪ NEISSERIA MENINGOCOCCUS ▪ STREPTOCOCCUS
▪ PNEUMOCOCCUS
▪ MICROCOCCUS
▪ SARCINA

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GRAM POSITIVE AND GRAM-NEGATIVE COCCI

GRAM POSITIVE
- divided on the basis of arrangement of cells

Vibrio: They are curved, non-flexible, Gram negative, highly motile

- e.g. Vibrio cholerae.

Spirilla: Consists of coiled, non-flexous motile cells

- e.g. Spirilum minus

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GRAM POSITIVE AND GRAM-NEGATIVE COCCI

BACILLI

GRAM POSITIVE GRAM NEGATIVE

MOTILE NON-MOTILE MOTILE NON MOTILE

▪ VIBRIO ▪ SHIGELLA ▪ B. ANTHRACOID ▪ CORYNEBACTERIUM
▪ SALMONELLA ▪ KLEBSHIELLA CLOSTRIDIUM ▪ MYCOBACTERIUM
▪ ESCHERECHIA COLI ▪ BRUCELLA ▪ LACTOBACILLUS
▪ PROTEUS ▪ PASTURELLA ▪ LISTERIA (MOTILE
▪ PROVIDENTIA ▪ HEMOPHILUS ONLY AT 22C)
▪ PSEUDOMONAS ▪ ACTINOMYCES
▪ CITROBACTER ▪ CLOSTRIDIUM
▪ ALCALIGEN FECALIS PERFRINGENS
▪ HAFNIA ▪ CLOSTRIDIUM
TETANI

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SYSTEMS USED TO CLASSIFY
BACTERIA

3. Adansonian or numerical classification

- this system determines the degrees of
relationship between strains by a statistical
coefficient- (score+1 if the character is present, –
1 if it is absent)
- the classification depends upon total
scores of as many similarities and differences as
can be observed
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 SYSTEMS USED TO CLASSIFY
BACTERIA
4. Biochemical classification
- N-acetyl muramic acid does not occur in eukaryotic cell
- prokaryotic cell wall membrane lacks sterols which typically occur in
the membrane of eukaryotic cells
- Gram-positive bacteria contain glycerol techoic acid and ribitol techoic
acid and some monosaccharides like rhamanose and arabinose

5. DNA composition
- measures G + C (guanine and cytosine) content of bacterial
preparation
- there is wide range of G + C components of DNA in bacteria, varying
from 25 to 80 moles percent in different generation

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 Identification of Bacteria
1. Microscopic examination
- helps to find out whether the bacteria is cocci,
bacilli, vibrio spirillum or spirochaete.
- Gram staining
a. Gram positive organisms
b. Gram-negative organisms
2. Motility
- Pathogenic cocci are non-motile
- Among Gram-negative: bacilli, salmonella, Escherichia coli, proteus, pseudomonas
Motile: Alcaligens fecalis, Vibrio cholerae
- Among Gram-positive: bacilli clostridia and bacillus are motile
- Hanging drop preparation, dark ground microscopy, phase contrast, electron
microscope help in their study

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 2. Negative staining bacteria are mixed
with dyes (India ink nigrosin) - background
1. Simple stains - watery gets stained leaving the bacteria
solution of single basic dye such contrastingly colorless. It is useful in
as, methylene blue or basic demonstration of bacterial capsule
fuchsin are used.
4. Differential stains impart different
3. Impregnation methods where bacterial cells colors to different bacteria or their
and appendages that are too thin and delicate structures. In a stained film, bacterial
cannot be seen under ordinary microscope. shape, arrangement and presence of
They are thickened by impregnation of silver other cells (pus cells) are noted. The
on the surface to make them visible under two commonly used differential
light microscope, e.g. demonstration of stains are Gram’s stain and acid fast
spirochaetes and bacterial flagella. stain

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Gram
Stain - named after the Danish scientist, Hans
Christian Gram
- He originally devised the staining technique
- the most widely used stain in microbiology
that differentiates bacterial species into two
large groups (Gram positive and Gram
negative) based on the physical and chemical
properties of their cell walls.
- Basic steps:
Applying a primary stain (crystal violet) to
a heat-fixed smear of a bacterial culture,
followed by the addition of a mordant (Gram’s
iodine),

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Ziehl-Neelsen (ZN) Stain or Acid Fast Stain
- first described by a bacteriologist, Franz Ziehl and a pathologist
Friedrich Neelsen
- It is a differential bacteriological stain used to identify acid fast
bacteria, especially Mycobacterium.
- reagents used are Ziehl-Neelsen carbolfuchsin (primary stain), acid
alcohol and methylene blue (counterstain).
- 5% sulphuric acid is used for staining Mycobacterium leprae and 20%
for Mycobacterium tuberculosis.

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 Ziehl-Neelsen (ZN) Stain or Acid Fast Stain
- positive smear typically contain pink colored, rod shaped bacteria that
are slightly curved, sometimes branching, sometimes beaded in
appearance, present singly or in small clumps against a blue background
of other cells.

- Acid fastness of a bacterium is attributed to the high content of lipids,

fatty acids and higher alcohols found in the cell wall.

- Mycolic acid, acid fast, waxy material is present in all acid fast bacteria.

- Apart from lipids, integrity of the cell wall also contributes to the acid
fastness of a bacterium.

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Difference Between Prokaryotic Cell and
Eukaryotic Cell
Character Prokaryotic Cell Eukaryotic Cell

I Nucleus

Nuclear membrane Absent Present

Nucleolus Absent Present
Histones Absent Absent
Chromosome One More
Mitotic division Absent Present
DNA Circular Linear

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 Difference Between Prokaryotic Cell and
Eukaryotic Cell

II Cytoplasm Prokaryotic Cell Eukaryotic Cell

Cytoplasmic streaming Absent Present

Cytoplasmic ribosome 70s 80s
Pinocytosis Absent Present
Mitochondria Absent Present
Lysosomes Absent Present
Golgi apparatus Absent Present
Endoplasmic reticulum Absent Present

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 Difference Between Prokaryotic Cell and
Eukaryotic Cell
Prokaryotic Cell Eukaryotic Cell

III Chemical composition

Sterol Absent Present

Muramic acid Present Absent
Diaminopimelic acid May be Absent Present

IV Miscellaneous

Diameter 1 µm 10 µm
Oxidative phosphory Cell Chlorolation site membrane plast
Cilia Absent Present
Pili Present Absent
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 BASIC LAB
EQUIPMENT

MICROSCOPE INSERT TITLE HERE

 TYPES OF
MICROSCOPES

SIMPLE
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MICROSCOPE
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