Classification of bacteria
11.03.2025
General Microbiology 2022/2023 batch
PT 1202
Classification and identification of organisms
• Two separate but inter-related processes.
Classification involves identification of groups of organism
• Share common properties
• Differ from other groups.
Identification entails assignation of an unknown organism
to a group within a scheme of classification.
>100 years classified according to their “Gram reaction”
• Named after Christian Gram (1884).
• Based on ability to retain a crystal violet-iodine
complex when treated with organic solvents (acetone
or alcohol).
• Gram-positive bacteria retain stain – purple / blue-black
• Gram-negative bacteria cannot retain the dye complex,
need to be counterstained with a red dye such as
carbol fuchsin - appear pink to red
Shape of bacteria to classify them – 3 basic shapes.
• Cocci, from the Greek kokkos - a berry
• Rod shaped (bacilli, from the Latin bacillus - a stick or rod)
• Spiral.
Gram stain
Note
• most used stain in bacteriology
• differentiates between bacteria
•Colour / shape / arrangement
STAIN Bright Field Microscopy – 100X (oil immersion objective)
Gram Stained Cells
Gram Positive Cocci Gram Positive Rods
STAIN Bright Field Microscopy – 100X (oil immersion objective)
Gram Negative Cocci Gram Negative Rods
Taxonomy
Science of classification, identification, and nomenclature.
Classification - organisms usually organized into
subspecies,
species,
genera,
families,
higher orders.
Taxonomy:
Domains of life
Prokaryotae
Carl Woese(1970’s)
Image from Scientific American blog
3 domains
Classification
Orderly arrangement of bacteria into groups.
Nothing inherently scientific about classification
Different groups of scientists may classify the same organisms
differently.
(Eg :- clinical microbiologists are interested in the serotype, antimicrobial
resistance pattern, and toxin and invasiveness factors in Escherichia coli,
whereas geneticists are concerned with specific mutations and plasmids.
Identification
Practical use of classification criteria to distinguish certain
organisms from others
Verify the authenticity or utility of a strain or a particular reaction
isolate and identify the organism that causes a disease
Nomenclature (naming)
means by which the characteristics of a species are defined and
communicated among microbiologists.
A species name should mean the same thing to all
microbiologists, yet some definitions vary in different countries or
microbiologic specialty groups.
Eg: the organism known as Clostridium perfringens in the United States is
called Clostridium welchii in England.
Bacterial Species
Distinct organism with certain characteristic features,
or
Group of organisms that resemble one another closely in the
most important features of their organization.
( In the past, unfortunately, there was little agreement about these criteria or
about the number of features necessary to distinguish a species. )
Species were often defined solely by such criteria as
host range, pathogenicity
ability to produce gas during the fermentation of a given sugar.
NOW
Generally accepted genetic criteria that can be used to define
species in all groups of bacteria.
Numerical taxonomy
• Determined on phenotype
• Large number of phenotypic tests used
– Morphology
– Biochemical
– Cultural characteristics …..
• Relatedness measured by degree of
similarity
Strain – isolate from one specimen
Problems:
• Only about 5-20% of genetic potential tested
• Easier to perform tests more frequently used
• Gene and reaction not a straight forward one
• other factors may affect phenotypic
characterization
• Same set of reactions cannot be used to
classify all organisms
Phylogenetic approach
• Comparison of genes
• Usually carried out in research laboratories
• Results compared with phenotypic characterization to select
tests which would give most reproducible and accurate results
– Genome size (measured in molecular weight)
– Guanine + cytosine content (25%-75%)
– DNA relatedness under conditions optimal for DNA reassociation
– Thermal stability of related DNA sequences
– DNA relatedness under supraoptimal conditions for DNA reassociation
DNA relatedness under conditions optimal for DNA
re-association
– Determined by allowing single stranded DNA from
one strain to associate with single stranded DNA
from another strain
• strains within a bacterial
species show 70– 100%
relatedness
•Relatedness between different
species is between 0-65%
•Relatedness is not = identical
Polyphasic approach
Thank You!
