677
such as those marketed by Difco. Heat-fi xed smears should be stained for 1 min
with crystal violet, washed in tap water, covered with Grams-iodine for 1 min, rewashed, decolorised by a few seconds in acetone-alcohol, and counterstained for
30 s in safranin. The smears are washed thoroughly, and gently blotted dry, prior to
microscopic examination preferably at a magnifi cation of ×1000.
• The acid-fast staining reaction
This reaction highlights the presence of Mycobacterium , Nocardia and possibly
Rhodococcus. Heat-fi xed smears may be fl ooded with carbol fuchsin, and heated
until the steam rises by means of wafting a source of heat (from a Bunsen burner or
cotton wool plug soaked with alcohol) underneath the slide. After 5 min, the stain is
washed away with tap water, and the smear decolorised with acid-alcohol until only
Table 13.1 (continued)
API ZYM test no.
Taxon (and source
of strains)
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Propionibacterium
acnes (CCM 3343)
− − − (+) − + − − − − − + − + − − − − + −
Ren. salmoninarum
(48 isolates)
− + − + − + − − + − + + − − − + − − (+) −
Rothia dentocariosa
(ATCC 17931)
− − − + − + + − + − − + − − − + − − − −
Sta. epidermidis
(NCIMB 2699)
− + (+) + − − − − − − + + − − − + − − − −
Str. agalactiae
(CCM 6187)
− + − + − − − − − − + + − − − (+) − − − −
Str. dysgalactiae
− + − − − + (−) − − − + (+) − − + + − − − −
Streptomyces
griseus (ATCC
23345)
− + − + − + − − − + − + − − − − − − − −
Presumptive
coryneform
(laboratory isolate
198)
− − + + − + − − − − + + − − − + − − − −
1 control, 2 alkaline phosphatase, 3 esterase (butyrate), 4 esterase (caprylate), 5 lipase (myristate),
6 leucine arylamidase, 7 valine arylamidase, 8 cystine arylamidase, 9 trypsin, 10 chemotrypsin, 11
acid phosphatase, 12 phosphoamidase, 13 α-galactosidase, 14 β-galactosidase, 15 β -glucuronidase, 16 α-glucosidase, 17 β -glucosidase, 18 N-acetyl- β -glucosaminidase, 19 α -mannosidase, 20
α -fucosidase
Sources: ATCC American Type Culture Collection, CCM Czechoslovak Collection of Microorganisms, NCDO National Collection of Dairy Organisms
NCIMB National Collection of Industrial and Marine Bacteria, NCTC National Collection of Type
Cultures; Dr. L.D. Ashburner, Freshwater Fisheries
Research Station, Victoria, Australia; Dr. B. Körmendy, Central Veterinary Institute, Hungary
a
15 ° C/18 h
b
Distinguish by results of the Gram-staining reaction
Phenotypic Tests
such as those marketed by Difco. Heat-fi xed smears should be stained for 1 min
with crystal violet, washed in tap water, covered with Grams-iodine for 1 min, rewashed, decolorised by a few seconds in acetone-alcohol, and counterstained for
30 s in safranin. The smears are washed thoroughly, and gently blotted dry, prior to
microscopic examination preferably at a magnifi cation of ×1000.
• The acid-fast staining reaction
This reaction highlights the presence of Mycobacterium , Nocardia and possibly
Rhodococcus. Heat-fi xed smears may be fl ooded with carbol fuchsin, and heated
until the steam rises by means of wafting a source of heat (from a Bunsen burner or
cotton wool plug soaked with alcohol) underneath the slide. After 5 min, the stain is
washed away with tap water, and the smear decolorised with acid-alcohol until only
Table 13.1 (continued)
API ZYM test no.
Taxon (and source
of strains)
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Propionibacterium
acnes (CCM 3343)
− − − (+) − + − − − − − + − + − − − − + −
Ren. salmoninarum
(48 isolates)
− + − + − + − − + − + + − − − + − − (+) −
Rothia dentocariosa
(ATCC 17931)
− − − + − + + − + − − + − − − + − − − −
Sta. epidermidis
(NCIMB 2699)
− + (+) + − − − − − − + + − − − + − − − −
Str. agalactiae
(CCM 6187)
− + − + − − − − − − + + − − − (+) − − − −
Str. dysgalactiae
− + − − − + (−) − − − + (+) − − + + − − − −
Streptomyces
griseus (ATCC
23345)
− + − + − + − − − + − + − − − − − − − −
Presumptive
coryneform
(laboratory isolate
198)
− − + + − + − − − − + + − − − + − − − −
1 control, 2 alkaline phosphatase, 3 esterase (butyrate), 4 esterase (caprylate), 5 lipase (myristate),
6 leucine arylamidase, 7 valine arylamidase, 8 cystine arylamidase, 9 trypsin, 10 chemotrypsin, 11
acid phosphatase, 12 phosphoamidase, 13 α-galactosidase, 14 β-galactosidase, 15 β -glucuronidase, 16 α-glucosidase, 17 β -glucosidase, 18 N-acetyl- β -glucosaminidase, 19 α -mannosidase, 20
α -fucosidase
Sources: ATCC American Type Culture Collection, CCM Czechoslovak Collection of Microorganisms, NCDO National Collection of Dairy Organisms
NCIMB National Collection of Industrial and Marine Bacteria, NCTC National Collection of Type
Cultures; Dr. L.D. Ashburner, Freshwater Fisheries
Research Station, Victoria, Australia; Dr. B. Körmendy, Central Veterinary Institute, Hungary
a
15 ° C/18 h
b
Distinguish by results of the Gram-staining reaction
Phenotypic Tests