Do the tests for denitrification( P. putida does not denitrify), gelatin
hydrolysis, flourescent pigment, and non-flourescent, diffusable pigment.
To test for:
denitrification- inoculate a fluid mineral salt + 0.5% glucose + 0.5%
nitrate. Incubate anaerobically at 30 degrees 2-4 days. I used a 4% agar
seal. Denitrifiers will show growth and gas production. P. putida is
not a denitrifier whereas many others are.
Gelatin hydrolysis- stab a gelatin deep. Incubate at 30 degrees 2-4
days. Cool to 20 degrees and check for liquefaction. P. putida does not
hydrolyze gelatin while many other species do.
pigmentation- inoculate a King B slant with your isolate, incubate at 30
degrees 2-4 days. Observe under a UV lamp. This should show any
flourescent pigment. Also check for diffusible pigments that do not
flouresce. P. putida should show flourescent pigment but no diffusible
pigment. There are three common species showing flourescent pigment.
King B agar
peptone 20.0g
Agar 15.0g
Glycerol 10.0g
Dipotassium phosphate 1.5g
Magnesium Sulfate 1.5g
Distilled water to 1000.0ml
Hope this helps,
Amy Lotto
aurelia at u.washington.edu
