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The major hydroxy fatty acids of cellular lipids in Flavobacterium meningosepticum and Flavobacterium sp. King's group IIb were identified as 2-hydroxy 13-methyltetradecanoic, 3-hydroxy 13-methyltetradecanoic, 3-hydroxy palmitic, and 3-hydroxy 15-methylhexadecanoic acids using gas chromatography-mass spectrometry and GC-mass fragmentography. The concentration of these hydroxy fatty acids comprised up to 30-40% of the total extractable and 20-30% of the bound lipid fatty acids, respectively. From the stability for mild alkaline hydrolysis, 2-hydroxy fatty acids seemed to be attached with ester linkage, and 3-hydroxy fatty acids with amide linkage. [ABSTRACT FROM AUTHOR]

We have recently isolated from the gingival pockets, periapical lesions and saliva some anaerobic gram-negative, black-pigmented rods. Many of these isolates exhibited phenotypic characteristics similar to Prevotella intermedia (Bacteroides intermedius). However, several of these isolates, although resembling P. intermedia in most of the phenotypic expressions, were capable of fermenting lactose, a biochemical characteristic atypical of P. intermedia. These atypical clinical isolates (strains capable of fermenting lactose) and isolates exhibiting more typical phenotypic characteristics (i.e., lactose nonfermenting) were definitively identified as P. intermedia by DNA-DNA hybridization using a photoprobe biotin method. Quantitative hybridization of clinical isolates with labeled DNA of P. intermedia-type strains (ATCC 25611 and ATCC 33563) showed that almost all the clinical strains isolated from disease sites of adults belonged to the ATCC 25611 group, whereas strains isolated from the saliva of children belonged to the ATCC 33563 group. These data, together with the phenotypic characterization of the isolates, suggested that P. intermedia is a heterogeneous species both pheno- typically and genetically. [ABSTRACT FROM AUTHOR]

Rapid genetic identification of 23 Legionella species was performed using microdilution plate hybridization. When an isolate appeared to be a strain of the Legionella species, its DNA was quickly extracted, labelled with photobiotin and distributed to microdilution wells where 23 different DNAs had previously been immobilized. After hybridization for 90 min at 45 °C, the microdilution well which emitted the maximum fluorescence was determined. The criterion for interpreting results of this genetic identification method is given. [ABSTRACT FROM AUTHOR]

The susceptibility of 3233 strains of Pseudomonas aeruginosa, isolated primarily in 2001, as agents of infection at 37 medical institutes with various specialties in seven regions of Japan (ranging from Hokkaido to Kyushu/Okinawa), to 18 antipseudomonal agents known to be active against P. aeruginosa was evaluated, in accordance with the National Committee for Clinical Laboratory Standards (NCCLS) guidelines. Of the 18 antipseudomonal agents, including some combinations of beta-lactamase inhibitors and antibacterial agents, ciprofloxacin had the lowest minimum inhibitory concentration (MIC)(50) (0.25 microg/ml) against P. aeruginosa, followed by meropenem, with an MIC(50) of 0.5 microg/ml. The MIC(50) of 7 of the examined antibacterial agents (ceftazidime, cefozopran, imipenem, biapenem, gentamicin, tobramycin, and levofloxacin) was between 1 and 2 microg/ml. Among the antipseudomonal agents tested, tobramycin showed the lowest MIC(90) (2 microg/ml), which was not significantly different from its MIC(50) (1 microg/ml). The MIC(90) of the other antibacterial agents examined ranged from 8 to 64 microg/ml and more. The susceptibility of the 3233 strains to the 12 antibacterial agents covered by the NCCLS guidelines was determined according to the standard method of the NCCLS guidelines. The frequency of strains resistant to meropenem, gentamicin, or tobramycin was relatively low (7.5%-8.3%). The frequency of strains showing intermediate to severe resistance to tobramycin was particularly low (8.0%). The frequency of strains resistant to aztreonam, imipenem, or levofloxacin was 16.7%-19.0%, about twice as high as the frequency of strains resistant to tobramycin. The susceptibility pattern of the 3233 strains (isolated from seven regions of Japan) to five antibacterial agents (ceftazidime, piperacillin, imipenem, gentamicin, and ciprofloxacin) was evaluated in relation to the regions from which they were isolated. The MIC(50) values of these antibacterial agents did not differ significantly among the regions. However, the MIC(90) values of ceftazidime and gentamicin were higher for strains isolated from the Kansai region than for strains isolated from other regions. The MIC(90) of ciprofloxacin was higher for strains isolated from the Tohoku, Kansai, and Kyushu/Okinawa regions than for strains isolated from other regions. Of the 3233 strains, 89 were classified as multiple-drug-resistant (imipenem, gentamicin, and ciprofloxacin) strains. Of these 89 strains, 42 were isolated from urine, 17 from sputum or pharyngeal mucus, 13 from pus, 8 from blood, 1 from cerebrospinal fluid, and 8 from other specimens. The frequency of multiple-drug-resistant strains was higher among strains isolated from the Tohoku and Kansai regions than in strains isolated from other regions.

Following cerebrating ceremony in 20 June 2002, for the completion of Hiuga Sun-Park Hot Spring Bath "Ofunade-no-Yu" facilities, Miyazaki Prefecture, Kyushu Island, 200 neighbors were invited each day to experience bathing on 20 and 21 June. The Bath "Ofunade-no-Yu" officially opened on 1 July 2002. On 18 July, Hiuga Health Center was informed that 3 suspected Legionella pneumonia patients in a hospital and all of them have bathing history of "Ofunade-no-Yu". Health Center officers notified Hiuga City, the main proprietor of the Bath business, that on-site inspection on sanitary managements will be done next day and requested the City to keep the bath facilities as they are. On 19 July, Health Center officers collected bath water from seven places and recommended voluntary-closing of "Ofunade-no-Yu" business. Because of various reasons, Hiuga City did not accept the recommendation and continued business up to 23 July. Because Legionella pneumophila serogroup 1 strains from 4 patients' sputa and several bath water specimens were determined genetically similar by Pulsed Field Gel Electrophoresis of Sfi I-cut DNA. "Ofunede-no-Yu" was regarded as the source of infection of this outbreak. On 24 July, "Ofunade-no-Yu" accepted the Command to prohibit the business. Among 19,773 persons who took the bath during the period from 20 June to 23 July, 295 became ill, and 7 died. Among them, 34 were definitely diagnosed as Legionella pneumonia due to L. pneumophila SG 1, by either one or two tests of positive sputum culture, Legionella-specific urinary antigen, and significant rise of serum antibody titer against L. pneumophila SG 1. In addition to the 8 items shown by Miyazaki-Prefecture Investigation Committee as the cause of infection. Hiuga City Investigation Committee pointed out following 3 items: 1) Insufficient knowledge and understanding of stuffs on Legionella and legionellosis; 2) Residual water in tubing system after trial runs might lead multiplication of legionellae in it; and 3) Inadequate disinfection and washing for whole circulation system prior the experience bathing. The Hiuga City Committee directed 24 measures to improve the sanitary condition of the facility including following 5 items. 1) Fix the manual for maintenance and management of the bath. 2) Keep sufficient overflow of bath water. 3) Put disinfection of filters into practice. 4) Precise measurement and control of the residual chlorine concentration in bath water. 5) Replacement of filtrating material from crushed porous ceramic into natural sand.

Growth characteristics including cell-arrangement of the type strain of Bacillus anthracis EY 3169T=ATCC 14578T grown on agar plates in level 3 laboratory were observed by both light and laser microscopes. Small daughter colonies appeared on parent colonies grown on 5% sheep blood or chocolate agar plates after 12 days incubation at room temperature. Daughter colonies, stained by Wirtz-Conklin method, were composed with vegetative cells and spores. Growth of daughter colonies might be supported by the debris of cells in the parent colony. Colonies grown under anaerobic conditions were flat with smooth edges, and the cells neither formed chains of any length, nor produced any spores after 25 days incubation at room temperature. It was thought that spores of B. anthracis were produced at the terminal stage of individual cell life instead of under unfavorable conditions for the organism. Air is needed for spore formation and cell-chain formation. More nutrients, probably amino acids, are needed for anaerobic growth rather than aerobic.

A polyphasic taxonomic study was performed on the phytopathogenic bacterial strains DAPP-PG 224(T) and DAPP-PG 228, which cause brown spot on yellow Spanish melon (Cucumis melo var. inodorus) fruits. Based on the presence of glucuronosyl ceramide (SGL-1) in cellular lipids, the results of fatty acid analysis and 16S rDNA sequence comparison, the strains had been identified as belonging to the genus Sphingomonas and as phylogenetically related to Sphingomonas mali, Sphingomonas pruni and Sphingomonas asaccharolytica. The levels of 16S rDNA sequence similarity of these three species to strain DAPP-PG 224(T) were respectively 98.0, 98.0 and 97.4%. DNA-DNA hybridization experiments between strains pathogenic on melon fruit and S. mali, S. pruni and S. asaccharolytica revealed < or = 16% relatedness. Based on these results, the two isolates studied are regarded as independent from the type strains of the three species mentioned above. Sphingomonas strains from melon fruits are recognized as forming a genetically and phenotypically discrete species and to be differentiated by phenotypic characteristics from all 29 named species of the genus. Thus, the name Sphingomonas melonis sp. nov. is proposed for the isolates from diseased melon fruits. The type strain is DAPP-PG 224(T) (= LMG 19484(T) = DSM 14444(T)). The G+C content of DNA of the type strain is 65.0 mol%.