Table 2
(continued)
Test
Inoculation
Incubation
Reagents
Positive reaction
Negative reaction
Growth on 5%
NaCl
Prepare a light suspension of
the test isolate from a pure
culture. Transfer 0.1 mL to
one L-J + 5% NaCl and one
control tube of NaCl-free
L-J medium
Incubate in 5–10% CO
2 at
35–37
C with caps
loosened. Check weekly for
up to 4 weeks
N/A
Growth
No growth
MacConkey
w/out
crystal violet
Inoculate 0.1 mL from a
growing test culture in
broth that is at least 7 days
old. Streak for isolation
Incubate in ambient air at
28–35
C for 5–11 days
N/A
Growth
No growth
Arylsulfatase
3 day/
14 day
Add one drop actively
growing culture in a liquid
medium to each tube
(culture should be at least
7 days old)
35–37
C in ambient air for
3 and 14 days
Six drops of 2 N sodium
carbonate
Red/pink
No color
development
Polysorbate
80
hydrolysis
Mix two drops of polysorbate
80 substrate to 1.0 mL of
sterile demineralized water.
Add one loopful of test
isolate from a young,
actively growing culture
slant
Aerobically at 35–37
C.
Examine at 1, 5, and
10 days. Do not shake
tubes while reading them
N/A
Red/pink
Amber color
Urea hydrolysis Inoculate a loopful of growth
from a young actively
growing culture into urea
broth. Vigorously mix
against bottom and sides of
tube
Aerobically at 33–37
C for
up to 7 days. Incubate an
uninoculated tube as a
negative control
N/A
Red/pink
No color change
18
Elizabeth Wallace et al.