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Laboratory Identification of Bacteria and Taxonomy

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19 views28 pages

Laboratory Identification of Bacteria and Taxonomy

Uploaded by

siva dharshini
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© © All Rights Reserved
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Laboratory Identification of

Bacteria and Taxonomy

Prepared by:
Dr. Fahmi Iqbal Rabbi
Associate Professor
Department of Microbiology, NEMC
Identification of Bacteria
• morphology of bacterial colony on solid
medium.
• growth in liquid medium.
• biochemical reactions.
• antigenic structures.
• animal pathogenicity.
• antibiotic sensitivity.
• typing of bacterial strains.
• rapid identification methods.
• molecular methods.
Morphology of Bacterial Colony on
Solid Medium
■ The colonies may be a few millimeters in size:
pinhead size (Staphylococcus aureus) or pinpoint
(streptococci).
■ The shape may be circular or irregular, and surface
of the colonies may be smooth, rough, or granular.
■ The colonies on the medium may be flat, raised, low
convex, convex, or umbonate.
■ The edge may be entire, lobate, crenated, undulate.
■ The colonies may be transparent, translucent, or
opaque.
■ Certain bacterial colonies are associated with
production of pigments and hemolysis around them.
Growth in Liquid Medium
• Nutrient broth and peptone water are
frequently used as liquid media for growth of
bacteria.

• For example, streptococci produce granular


deposits t the bottom of the liquid medium,
whereas most of the Gram-negative bacteria
produce uniform turbidity.
Biochemical Reactions
• Catalase test
• Oxidase test.
• Indole test
• Carbohydrate fermentation test
• Oxidation–fermentation test
• Kligler’s iron agar/triple sugar iron agar test
• Urease test
• Citrate test
• Hydrogen sulfide production
• Voges–Proskauer test
Antigenic Structures

• Agglutination of biochemically confirmed


bacteria with specific antisera facilitates further
identification of the isolated bacteria.
Agglutination test is used in the identification of
presumptive isolates of pathogens (e.g.,
Salmonella) from clinical samples. It is also used
for the grouping of beta hemolytic streptococci.
Animal Pathogenicity
• Some pathogenic bacteria and their bacterial
metabolites produce characteristic lesions in
laboratory animals. The most commonly employed
animals are rats, guinea pigs, mice, and rabbits.

• For example, guinea pigs are commonly used for


performing animal pathogenicity of C. diphtheriae,
Clostridium perfringens, and M. tuberculosis.
Antibiotic Sensitivity
• Determination of antibiotic sensitivity of an
isolate from a patient is essential for the
choice of drug therapy. In some cases,
sensitivity of an organism to a particular agent
helps in the identification, e.g., Streptococcus
pyogenes is sensitive to bacitracin and
Streptococcus pneumoniae to optochin.
Typing of Bacterial Strains
• Phenotypic techniques :
■ Biotyping
■ Serotyping
■ Bacteriocin typing
■ Phage typing
• Genotypic techniques : Genotypic techniques
depend on differences related to the genome
of bacteria.
Rapid Identification Methods

• Latex particle agglutination, coagglutination,


direct fluorescent antibody test, and dot
enzyme-linked immunosorbent assay (ELISA) are
the most frequently used techniques in the
clinical laboratory for rapid detection of
microbial antigens directly in clinical specimens.
Molecular Methods
• Molecular methods, such as nucleic acid probes,
polymerase chain reaction (PCR), and other
amplification procedures are also used
increasingly nowadays for identification of
microorganisms. Genetic probes are based on
the detection of unique nucleotide sequences
with the DNA or RNA of a microorganism.
Bacterial Taxonomy

(a) Bacterial classification of organism and

(b) Nomenclature or naming of the microbial


isolates.
Bacterial Classification

• Bacterial classification may be defined as the


arrangement of organisms into taxonomic groups
(taxa) on the basis of their phenotypic
(observable) and genotypic (genetic) similarities
and differences. It allows proper and systematic
grouping of microorganisms.
Three-domain System

• The classification scheme currently favored by


most microbiologists is the three-domain
system. This designates all organisms as
belonging to one of the three domains—
Bacteria, Archaea, and Eucarya.
Five-kingdom System

• The five kingdoms in this system are the


Plantae, Animalia, Fungi, Protista (mostly
single-celled eukaryotes) and Prokaryotae.
• Organisms are classified into three main
kingdoms: Animals, Plants, and Protista.

• The Protista contains unicellular microorganisms


including eukaryotes and prokaryotes.
Phylogenetic classification
It denotes an evolutionary arrangement of species.

• Division: Protophyta
• Class: Schizomycetes
• Order: Eubacteriales
• Family: Enterobacteriaceae
• Tribe: Escherichiae
• Genus: Escherichia
• Species: coli
• Some characters of special importance, such
as Gram staining properties, lactose
fermentation, spore formation, etc., are used
to differentiate major groups, whereas less
important properties, such as nutritional
requirements for growth of bacteria,
production of certain enzymes by bacteria,
etc., are employed to distinguish minor
groups, such as the genera and species.
Adansonian or Numerical Classification

• The Adansonian or numerical classification, so


called after Michael Adanson who introduced
it in the eighteenth century, avoids the use of
weighted characteristics.
Intraspecies Classification
• It is often necessary to subclassify bacterial
species for diagnostic or epidemiological
purposes on the basis of biochemical
properties (biotypes), antigenic features
(serotypes), bacteriophage susceptibility
(phage types) or production of bacteriocins
(colicin types).
Genetic classification

• The genetic or molecular classification is


based on homology of the DNA base
sequences of the microorganisms.
Nomenclature of Microorganisms
• Nomenclature refers to the naming of
microorganisms.

• The nomenclature of microorganisms is


governed by the International Committee on
Systematic Bacteriology and published as
Approved List of Bacterial Names in the
International Journal of Systematic Bacteriology.
• The nomenclature and classification of viruses
are governed by the International Committee
on Taxonomy of Viruses.
Two kinds of names are usually given to bacteria—
common name and scientific name:

• The common or casual name for a


microorganism varies from country to country
and is usually known in the local language. For
example tubercle bacillus, typhoid bacillus,
gonococcus are common names for
communication at the local level.
• The scientific name is the international name
that is accepted throughout the world. By
accepted taxonomic conventions, the order
names end in ales (e.g., the order
Eubacteriales), family names end in aceae
(e.g., the family Enterobacteriaceae), and the
tribe names end in eae (e.g., the tribe
Proteae).
• The scientific name of the bacterium when
written for the first time, is written in full (e.g.,
Escherichia coli), but later mentioned in an
abbreviated form (e.g., Escherichia coli).

• When bacteria are referred to as a group,


their names are neither capitalized nor
italicized or underlined (e.g., streptococci).

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