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3 Table 1. Type strains used for DNA-DNA hybridization ATCC: American Type Culture Collection, Maryland. CCM: Czechoslovak Collection of Microorganisms, Purkyne University of Brno, Brno. DSM: Deutsche Sammlung von Mikroorganismen und Zellkulturen Gottingen. GIFU: Department of Microbiology, Gifu University School of Medicine, Gifu. JCM: Japan Collection of Microorganisms, Saitama. NCTC: National Collection of Type Cultures, London. TCC: Terumo Culture Collection, Kanagawa.
4 Fig. 1 Size and substrate arrangement of STAPHYOGRAM plate
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6 Fig. 2 DNA-DNA homology among the 30 strains of aerobic, Gram-positive and catalase-positive cocci. DNA from each type strain for the 30 species of aerobic Gram-positive and catalase-positive cocci was labeled with photobiotin, and fluorometric hybridization were carried out against the purified DNA of each type strain fixed in well. Numbers on the vertical axis indicated homology values, and strains in well number 1 through 31 (horizontal axis) are in order shown in Table 1.
7 Fig. 2 Continued
8 Table 2. Different profile number obtained from STAPHYOGRAM plate culture among the type strain of 30 species of three genera
9 Table 3. Identification of aerobic Gram-positive, catalase-positive cocci by microplate hybridization in comparison with those by conventional method Table 4. Frequency of STAPHYOGRAM profiles a): Species identity was determined by DNA-DNA hybridization.
10 Table 5. Result of STAPHYOGRAM additional tests a): Species identified by DNA-DNA hybridization b): COAL: Coagulase HEM: Hemolysis (bovin) NOVR: Novobiocin resistance RIB: Acid from ribose PAL: Alkaline phosphatase CEL: Acid from cellobiose PIG: Pigment ORN: Ornithine decarboxylated Table 6. Number of strains required additional tests for identification
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12 1) Baird-Parker, A.C. (1967): International committee on nomenclature of bacteria subcommittee on taxonomy of staphylococci and micrococci. Int. J. Syst. Bacteriol., 17, ) Christensen, G.D., Simpson, W.A., Bisno, A.L., and Beachey, E.H. (1982): Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infect. Immun., 37, ) Darbyshire, P.J., Weightman, N.C., and Speller, D.C.E. (1985): Problems associated with indwelling central venous catheters. Arch. Dis. Child., 60, ) Devriese, L.A., Poutrel, B., Kilpper-Bali, R., and Schleifer, K.H. (1983): Staphylococcus gallinarum and Staphylococcus caprae, two new species from animals. Int. J. Syst. Bacteriol., 33, ) Ezaki, T., Hashimoto, Y., and Yabuuchi, E. (1989): Fluorometric deoxyribonucleic acid-deoxyribonucleic acid hybridization in microdilutin wells as an alternative to membrane filter hybridization in which radioisotopes are used to determine genetic relatedness among bacterial strains. Int. J. Syst. Bacteriol., 39, ) Falk, D. and Guering, S.J. (1983): Differentiation of Staphylococcus and Micrococcus spp. with the Taxo A bacitracin disk. J. Clin. Microbiol., 18, ) Igimi, S., Takahashi, E., and Mitsuoka, T. (1990): Staphylococcus schleiferi subsp. coagulans subsp. nov., isolated from the external auditory meatus of dogs with external earotitis. Int. J. Syst. Bacteriol., 40, ) Judicial commission opinion 17. (1958): Conservation of the generic name Staphylococcus Rosenbach, designation of Staphylococcus aureus Rosenbach as the nomenclatural type of the genus Staphylococcus aureus Rosenbach, and designation of a neotype culture of Staphylococcus aureus. Int. Bull. Bacteriol. Nomencl. Taxon. 8, ) Kloos, W.E. and Schleifer, K.H. (1975): Simplified scheme for routine identification of human Staphylococcus species. J. Clin. Microbiol., 1, ) Kocur, M., Pacova, Z., and Martinec, T. (1972): Taxonomic status of Micrococcus luteus Cohn 1872 and designation of the neotype strain. Int. J. Syst. Bacteriol., 22, ) Macfaddin, J.F. (1980): Alkaline phosphatase, heat-resistant tests, 1-4, In Biochemical tests for identification of medical bacteria, Second edition, Baltimore, Williams & Wilkins Co. 13) Macfaddin, J.F. (1980): Decarboxylase tests and dehydrolase tests, 78-93, In Biochemical tests for identification of medical bacteria, Second edition, Baltimore, Williams & Wilkins Co. 14) Moller, J.K. (1988): Observation on multiple drug resistance in coagulase-negative staphylococci isolated in hospitals from 1975 to J. Hosp. Infec., 11, ) Moller, V. (1955): Simplified tests for some amino acid decarboxylases and for the arginine dihydrolase system. Acta Pathol. Microbiol. Scand., 36, ) Poutrel, B. and Caffin, J.P. (1981): Lysostaphin disk test for routine presumptive identification of staphylococci. J. Clin. Microbiol., 13, ) Schleifer, K.H. (1986): Family Micrococcaceae , In Krieg, N.R. and J.G. Holt (eds), Bergey's manual of systematic bacteriology Vol.2, Baltimore, Williams & Wilkins Co. 18) Schleifer, K.H., Kloos, W.E., and Moore, A. (1972): Taxonomic status of Micrococcus luteus Cohn 1872: correlation between peptidoglycan type and genetic compatibility. Int. J. Syst. Bacteriol., 22, ) Schleifer, K.H. and Kocur, M. (1973): Classification of staphylococci based on chemical and biochemical properties. Arch, Microbiol., 93, ) Skerman, V.D.B., McGowan, V., and Sneath,
13 P.H.A. (1980): Approved lists of bacterial names. Int. J. Syst. Bacteriol., 30, ) Subcommittee on taxonomy of staphylococci and micrococci. Recommendations. (1965), Int. Bull. Bacteriol. Nomencl. Taxon., 15, ) Varaldo, P.E., Kilpper-Balz, R., Biavasco, F., Satta, G., and Schleifer, K.H. (1988): Staphylococcus delphini sp. nov., a coagulasepositive species isolated from dolphins. Int. J. Syst. Bacteriol., 38, ) Wayne, L.G., Brenner, D.J., Colwell, R.R., Grimont, P.A.D., Kandler, O., Krichevsky, M.I., Moore, L.H., Moore, W.E., Murray, R.G.E., Stackbrandt, E., Starr, M.P., and Truper, H.G. (1987): Report of the Ad Hoc committee on reconcilation of approaches to bacterial systematics. Int. J. Syst. Bacteriol., 37,
14 STAPHYOGRAM, a new rapid identification kit for the aerobic, Gram-positive, catalase-positive cocci-application of fluorometric microplate hybridization for the pre-identification of 386 isolates used Yoshiaki KAWAMURA1), Kouji NAGASE2), Masakazu ENDO2), Takashi SHIGEMATSU2), Takayuki EZAKI1) and Eiko YABUUCHI1) 1) Department of Microbiology, Gifu University School of Medicine, Gifu 500 2) Diagnostic Products Department, R & D Center, Terumo Corporation, Ashigarakami-gun, Kanagawa , Japan A new simplified test kit, STAPHYOGRAM plate, was developed for 4-hr identification of aerobic, Gram-positive and catalase-positive cocci. The plate has 18 wells, in which different dehydrated substrates and nutrients are fixed. An 18-hr agar-culture suspension of a test strain with a turbidity of McFarland No.4 was distributed into all wells in 50-ƒÊl quantities. After 4-hr incubation at 37C, the profile number was obtained by summarizing positive reactions. The ability of the plate to differentiate the type strains of the 30 species of the three genera in the family Micrococcaceae was confirmed. These three genera are Staphylococcus, Micrococcus and Stomatococcus. The applicability of the fluorometric microplate hybridization technique to identification of aerobic, Gram-positive and catalase-positive cocci was confirmed by homologous hybridization among the type strains of the 30 species. Thus, 386 isolates of human and animal origin were pre-identified by microplate hybridization and used for evaluating the STAPHYOGRAM plate. Of the 236 profile numbers thus obtained with the 386 isolates, 218 (92.4%) were speciesproper each and all for the 15 species of Staphylococcus and Stomatococcus mucilaginosus. A total of 342 (88.6%) of the 386 isolates were given such profile numbers, and were identified without any additional test. Among the 15 species identified primarily by the results of STAPHYOGRAM plate culture, S. caprae, S. lugdunensis, S. gallinarum and S. delphini were validly published after Approved Lists of Bacterial Names. The identified strains of S. caprae (48), S. haemolyticus (46), S. capitis (35) numbered between those of S. epidermidis (67) and S. saprophyticus (31). Profile numbers common to two species were seven (27 strains) and that to four species was one (17 strains). These 44 strains were identified with one to three additional tests. From these results, we were convinced that the STAPHYOGRAM test plate is useful for the rapid identification of members of family Micrococcaceae. By compiling STAPHYOGRAM plate data on genetically identified strains, an exclusive list of profile numbers will soon be prepared for perfection of the kit. Present address: Yoshiaki KAWAMURA2) Eiko YABUUCHI Department of Bacteriology, Osaka City Univ. Medical School, Abeno-ku Osaka 545, Japan
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CHLAC 1 2 3 3,. (CHLAC), 1).,.,, CHLAC,.,. Suspicious Behavior Detection based on CHLAC Method Hideaki Imanishi, 1 Toyohiro Hayashi, 2 Shuichi Enokida 3 and Toshiaki Ejima 3 We have proposed a method for
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2011 Vol.60 No.4 p.332 338 Usefulness of regional education program for dietary salt reduction: Self-monitoring of urinary salt excretion Kenichiro YASUTAKE[1] Kayoko SAWANO[1] Shoko YAMAGUCHI[1] Hiroko