Upload
aubrey-dawson
View
215
Download
2
Embed Size (px)
Citation preview
Biochemical Characterization of Bacteria
LECTURE 8:
Microbiology and Virology; 3 Credit hours
Atta-ur-Rahman School of Applied Biosciences (ASAB)National University of Sciences and Technology (NUST)
Catalase testo Principle:• Tests the ability of some bacteria to produce catalase
enzyme.• Procedure:• Pick up the test colony on a platinum loop and immerse it in
few drops of 3 % H2O2 (hydrogen peroxide).• Interpretation:• Rapid bubbling indicates oxygen production and a positive
test.• This test is used to differentiate between staphylococci (+Ve)
and streptococci (-Ve).
Catalase Test
Coagulase test
• Detects the coagulase bound to the organism.• Procedure:• Homogenous suspension of the test organism is
made in a drop of saline on a slide then mixed with a drop of undiluted human or rabbit plasma.
• Staphylococcus aureus clumps within 15 sec. because coagulase enzyme precipitates the fibrin in the plasma on the cell surface.
Coagulase test
Oxidase test
• Principle:• Some bacteria produce oxidase enzyme which reduces the
oxidase reagent (tetramethyl-p-phenylene-diamine hydrochloride) to a deep purple color.
• Procedure:• Done by picking up a portion of the colony tested and
smearing it on a filter paper impregnated with oxidase reagent.• Interpretation:• The immediate development of a deep purple color indicates
positive test.• Examples: Neisseria & Pseudomonas & Vibrios.
Oxidase test
H2S test
• Principle:• Production of H2S from proteins.• Procedure:• Inoculate the organism on peptone water.• After incubation, put lead acetate paper at the mouth
of the tube.• Interpretation:• If the organism produces H2S, a black color is formed.• Examples: Proteus & Salmonella typhi.
H2S test
• Tests for enzyme cysteine desul-furase bacteria with this enzyme can break down amino acid cysteine
• Growth medium- SIM medium agar • Positive= black• Negative= medium remains yellow or black
line forms just below surface
H2S test
Hydrogen Sulfide (H2S) TestPositive=blackNeg= yellow or yellow with black line