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The basonym of Shigella dysenteriae (Shiga 1898) Castellani and Chalmers 1919 was first and officially proposed in 1897 as Bacillus dysentericus (sic) in Saikingaku Zasshi, published in Japan. In accordance with Section 6 (Citation of Authors and Names) of the Bacteriological Code (1990 Revision), the year of the first proposal of Shigella dysenteriae has to be corrected from Bacillus dysenteriae 1898 to Bacillus dysentericus (sic) 1897.

A 57-year-old man presented with pneumonia, respiratory distress, and myelodysplastic syndrome. A diagnosis of Legionnaires' disease due to Legionella pneumophila (L. pneumophila) was established. The patient had long been drinking tap water via a conduit from a hot spring resource, from which L. pneumophila was also isolated. Both the patient's strain and the water strain of L. pneumophila were identified as serogroup 1, and the genetic relatedness between the two strains as seen by pulsed-field gel electrophoresis was 87%. The patient was successfully treated with erythromycin, fluoroquinolone, and rifampicin. This case raises an important issue on public health represented by legionellosis in Japan.

A 56-year-old Japanese male was admitted to Toyohashi Municipal Hospital because of fever, cough, and dyspnea. Chest X-ray film showed bilateral alveolar infiltrates. He suffered from severe hypoxemia and was given a diagnosis of acute respiratory distress syndrome. He was also complicated with disseminated intravascular coagulation and pseudomembranous colitis. He fully recovered by intensive treatment with antibiotics, mechanical ventilation and endotoxin eliminating therapy. Legionella longbeachae was isolated from his respiratory specimens and was regarded as the etiologic agent of his pneumonia.

A novel sphingoglycolipid was isolated from Sphingomonas yanoikuyae, and its structure was identified as a galacturonosyl-beta (1-->1)-ceramide. This was a characteristic sphingoglycolipid present in S. yanoikuyae and certain other species of Sphingomonas, such as Sphingomonas mali, Sphingomonas terrae, and Sphingomonas macrogoltabidus, but not in the type species of Sphingomonas, Sphingomonas paucimobilis.

A polar multitrichous gram-negative motile rod, EY 3383, originally identified as Burkholderia thailandensis, revealed a DNA-DNA reassociation rate of 36.7%, under stringent conditions, with the type strain of B. thailandensis, despite the 16S rDNA homology value between two type strains being as high as 97.9%. The strain was clearly differentiated from the type strain of B. thailandensis by physiological, bio-chemical, and nutritional characteristics, without significant difference in cellular fatty acid and lipid composition. Based on the results of 16S rDNA sequence analysis, DNA-DNA hybridization and phenotypic characterization, Burkholderia uboniae sp. nov. is herein proposed. The type strain is NCTC 13147=EY 3383, isolated on 8 December 1989 from surface soil along the roadside in Ubon Ratchathani, Thailand. Major respiratory quinone is ubiquinone-8(Q8). G+C content of DNA is 69.71%.

Based on the results of phylogenetic analysis of the 16S rDNA sequences and the presence of N-2'-hydroxymyristoyl dihydrosphingosine 1-glucuronic acid (SGL-1) and 2-hydroxymyristic acid (non-hydroxymyristic acid in Zymomonas) in cellular lipids, a new family, Sphingomonadaceae, for Group 4 of the alpha-subclass of the class Proteobacteria is herein proposed and a description of the family is given. The family consists of six genera, Sphingomonas, Erythrobacter, Erythromicrobium, Porphyrobacter, Sandaracinobacter and Zymomonas. Thus, all the validly published and currently known genera in Group 4 of the alpha-subclass of Proteobacteria belong to Sphingomonadaceae fam. nov. Among them, type strains of two species, Porphyrobacter and Erythrobacter, Sandaracinobacter sibiricus and Sphingomonas ursincola, respectively, are facultatively photosynthetic due to bacteriochlorophyll (Bchl)-a. The type strains of two subspecies of Zymomonas mobilis are facultative anaerobes. The presence of SGL-1 together with the results of a phylogenetic analysis of the 16S rDNA sequences recommends a new criteria by which to define the new family Sphingomonadaceae. The type genus is Sphingomonas Yabuuchi et al. 1990.

We have evaluated urine specimens of presumptive cases of legionnaires' disease (110 cases, 173 sample), collected in the past eight years (April, 1990-August, 1998) with the Binax EIA kit which detects the soluble antigen of Legionella pneumophila serogroup (SG) 1, and the Biotest EIA kit which detects Legionella species. Seven cases (19 specimens) were positive for the Binax EIA kit, and nine cases (22 specimens) were positive for the Biotest EIA kit. The sensitivity for culture, PCR, IFA method were 100%, 100%, and 50%, the specificity for these method were 93%, 97.1%, and 90% respectively. Overall agreements for these method were 93.5%, 97.4%, 86.8%, these results suggested that the urinary antigen detection test had high sensitivity and specificity. Our study indicated that concentrated urine samples increase sensitivity. We also evaluated the capabilities of both EIAs to detect soluble antigens were extracted from bacterial suspension of 18 strains of 5 Legionella species by heating. Both assays detected L. pneumophila serogroups 1 to 14, L. bozemanii. The Binax EIA proved to be useful as the Biotest EIA for diagnosis of legionellosis caused by Legionella species and serogroups other than L. pneumophila serogroup 1. Some cases have been shown to excrete antigen for prolonged period of times despite recovery from infection, so that the patient's history should be sought. The urine antigen detection EIA methods proved to be rapid and easy to use, detect antigen in the early stage of the disease with high sensitivity and specificity. Its use for the definition of legionellosis should be considered in Japan.

Based on the results of a phylogenetic analysis of 16S rRNA and the presence of sphingoglycolipid in cellular lipids of the type strains, transfer of "Rhizomonas" suberifaciens, Blastomonas natatoria and Erythromonas ursincola to the genus Sphingomonas as Sphingomonas suberifaciens (van Bruggen et al 1990) comb. nov., Sphingomonas natatoria (Sly 1985) comb. nov., and Sphingomonas ursincola (Yurkov et al 1997) comb. nov. are herein proposed together with the emendation of genus Sphingomonas. The type strain of S. suberifaciens is van Bruggen Cal=ATCC 49382=NCPPB 3629=IFO 15211=JCM 8521, that of S. natatoria is ATCC 35951 =DSM 3183=NCIMB 12085=JCM10396, and that of S. ursincola is DSM 9006= KR-99.

Microbiological contamination of hot spring bath water is a public health concern. A province-wide survey was carried out to determine the extent and distribution of both Legionella and free-living amoebae contamination. Among 30 samples of hot spring bath from 12 sites in Kanagawa, Japan, L. pneumophila was detected in 21 water samples from 11 sites, ranging from 10(1)-10(3) CFU/100 ml. Serogroups 3, 5 and 6 of L. pneumophila were predominantly isolated from the samples. Naegleria (46.7%), Platyamoeba (33.3%), Acanthamoeba (10.0%) and 2 other genera of free-living amoebae were detected in 22 samples from 11 sites. One or more genera of host amoebae of Legionella occurred in 17 samples (56.7%) from 9 sites. Another thing to be noted is that 13 water samples contained N. lovaniensis. Although N. lovaniensis is nonpathogenic, it is considered an indicator organism for places that are suitable for the growth of N. fowleri, a causative agent of primary amoebic meningoencephalitis in man.

Based on the results of GC content determination and 16S rRNA sequence analysis among the type strains of Achromobacter xylosoxidans, 4 Alcaligenes species, 5 Bordetella species, and 12 species of 4 other genera, the separation of genus Achromobacter Yabuuchi and Yano 1981, with the type species Achromobacter xylosoxidans, is confirmed. Alcaligenes ruhlandii (Packer and Vishniac) Aragno and Schlegel 1992 is a distinct species and not a senior synonym of Achromobacter xylosoxidans. Alcaligenes ruhlandii and Alcaligenes piechaudii Kiredjian et al 1986 are transferred to genus Achromobacter. Thus 2 new combinations, Achromobacter ruhlandii (Packer and Vishniac) and Achromobacter piechaudii (Kiredjian et al) are proposed; their type strains are ATCC 15749 and ATCC 43552, respectively. Alcaligenes denitrificans Rüger and Tan 1983 is also transferred to genus Achromobacter and ranked down to the subspecies of Achromobacter xylosoxidans. Thus a new subspecies name, Achromobacter xylosoxidans subsp. denitrificans (Rüger and Tan) is proposed. The type strain of the subspecies is ATCC 15173. This proposal automatically creates type subspecies, Achromobacter xylosoxidans subsp. xylosoxidans, with type strain ATCC 27061. An emended description of genus Achromobacter and of type species Achromobacter xylosoxidans are given.

In order to promote the serological diagnosis of legionellosis in the clinical laboratory, the cutoff values of serum anti-Legionella antibody titers for microplate agglutination (MPAT) test were determined. Antibody levels were tested for 178 serum specimens including 98 healthy persons, 22 ordinally workers having either metabolic or renal failure, definitely diagnosed patients of 17 mycoplasmal and 9 chlamydial pneumonia, 32 patients of other bacterial pneumonia. Heat killed unstained cell suspension of each strain of Legionella pneumophila serogroup (SG) 1a, 1b, SGs 2 to 6, L. bozemanii, L. dumoffii, L. gormanii, and L. micdadei were used as antigens. Strains of L. pneumophila SG 1b were mainly isolated from environmental specimens. However, in some Legionella pneumonia cases, etiologic agents were determined as L. pneumophila SG 1 b. Thus the representative strain of SG 1b was used as an antigen for the determination of the patient's antibody titer. Quantitative agglutination was performed by using a 96-well U-bottom microplate for each antigen. Test sera were diluted from 1:16 to 1:256. Results were read after 20 h at 25 degrees C. Cutoff values for 11 antigens were determined, at this moment, as 4-fold or greater increase in level to > or = 1:128 in paired sera, and > or = 1:256 in single serum. However, final diagnosis should be given by over-all coordination of serological results and clinical symptoms together with other laboratory findings. Two culture-positive Legionella pneumonia cases due to either SG3 or 6 in which significant rise of serum antibody titers against organisms of corresponding SG estimated by MPAT method were discussed.

To determine the infection source of a sporadic Legionella pneumonia case associated with a hot spring bath, we used five molecular methods, including repetitive element polymerase chain reaction (rep-PCR), arbitrarily primed PCR (AP-PCR), ribotyping, restriction endonuclease analysis (REA), and macrorestriction endonuclease analysis (MREA) by pulsed-field gel electrophoresis. L. pneumophila serogroup (SG) 3 strain EY 3702, isolated from an intratracheal specimen of a 71-year-old Japanese female who developed pneumonia after nearly drowning in a hot spring spa bath, produced rep-PCR and AP-PCR fingerprints identical to those of L. pneumophila SG 3 strains EY 3768 and EY 3769 isolated from the bath water. Four epidemiologically unrelated L. pneumophila SG 3 strains showed different rep-PCR or AP-PCR fingerprints from those of the three EY strains (EY 3702, 3768, and 3769). The three EY strains were also genotypically indistinguishable by ribotyping with EcoRI and PstI, by REA with EcoRI or HindIII, and by MREA with NotI. Based on these results, we identified the bath water of the hot spring spa as the source of infection of this patient, even though the viable number of the organisms in the bath water was low (3 CFU/100 ml) when determined 27 days after her nearly drowning.

The serodiagnosis of melioidosis is commonly performed with tests using protein or polysaccharide as antigen. However, due to the low sensitivity, specificity and difficulty in the preparation of the antigens, more simple, precise and reproducible diagnostic tests were required. A purified glycolipid antigen (GL) which is a specific lipid component of Burkholderia pseudomallei has been used in an ELISA. With this antigen, specific immunoglobulin G (IgG) was detected in 49 out of 50 melioidosis sera. IgG was also detected in 2 out of 185 (Japanese) and 16 out of 181 (Vietnamese) control sera. Thus, the sensitivity was 98.0%, and specificity was 98.9% and 91.1% in the Japanese and Vietnamese sera, respectively. When the ELISA and indirect haemagglutination (IHA) tests were combined, a sensitivity of 100% and specificity of 97.8% were achieved. The advantages of the glycolipid antigen are ease of preparation, stability, high sensitivity and specificity.

A 71-year-old Japanese female, was found unconscious by drawing, in a hot spring spa, at around noon of 20 October 1994. She recovered by emergency cardiopulmonary resuscitation, and admitted to the Takinomiya General Hospital, with adult respiratory distress syndrome (ARDS). Although she recovered from ARDS within 4 days after her admission, she developed severe pneumonia accompanied with the second attack of ARDS. Ordinary bacteriological culture of her respiratory specimens failed to yield any significant pathogen for her pneumonia, and neither cefazolin nor imipenem/cilastatin was effective. Thus minocyclin was given on the 7th hospital-day and this was effective for blood gas and C-reactive protein (CRP) levels. Intratracheal exsudate inoculated on BCYE alpha agar plate yielded grayish white colonies. Cells of the colonies were clearly agglutinated by anti-Legionella pneumophila serogroup (SG) 3 serum. Antibody titers of patient's paired sera against the strain L. pneumophila SG3 Bloomington-2 and the patient's strain (Y-1) were determined by microplate agglutination test, and a significant rise from 1:20 to 1:320 was demonstrated. Patient recovered by erythromycin treatment and was discharged on the 59th hospital day. L. pneumophila SG3 organisms were again isolated from the spa water where the patient drawn. From these findings described above, we diagnosed the patient as pneumonia due to L. pneumophila SG3, and the spa water was the most probable source of infection.

In August 1994, an epidemic of acute febrile illness occurred at the Education Center Building of a company in Shibuya-ku, Tokyo. All 43 trainees attended in two groups and 2 staff members of the Center fell ill. The 45 patients came to one of our hospitals in two groups, and 35 patients were treated. The patients were 4 males and 31 females, and the average age was 29.0 years. The duration until falling ill was 36 to 90 hours after entering the Center. Symptoms were fever, lumbago arthralgia, headache, dyspnea, general fatigue, etc. Physical examination revealed slightly injected mucosa of the pharynx in a patient who complained of a sore throat. On laboratory examination, leukocytosis with a left shift of the nucleus and elevation of serum CRP levels were found. Erythromycin (600 mg, daily) and nonsteroidal antiinflammatory drugs (NSAIDs) were given by mouth to almost every patient. Two patients were hospitalized. The illness was self-limited, generally lasting from two to five days. Strains of legionellae isolated from the water of the cooling tower located at the top of the Center, were identified as L. pneumophila serogroup 7. Since seroconversion in a patient against the cooling tower strain from 1:16 to 1:256 was determined and the clinical courses agreed with the definition of Pontiac fever by Glick et al, we concluded that the epidemic was an outbreak of Pontiac fever due to L. pneumophila serogroup 7. Pontiac fever is considered to be one of the community-acquired diseases. Thus, we have to note that Pontiac fever may be misdiagnosed as we examine patients who complain of the symptoms noted above.

From August 20 to 22, 1994, an outbreak of acute febrile illness occurred in a Training Center building of a company in Shibuya-ku, Tokyo. All 43 trainees attended in two groups and 2 Center staffs were attacked. Illness was self- limiting, generally lasting three days. Though strains of legionellae, isolated from the water of the cooling tower located at the top of the building, were identified as Legionella pneumophila by microplate DNA-DNA hybridization, they failed to agglutinate with antisera against L. pneumophila serogroups 1 through 6. Two strains were sent to the Centers for Disease Control, Atlanta, Georgia, USA, and determined as serogroup 7 of the species. Since the clinical courses agreed with the definition of Pontiac fever by Glick et al. and seroconversion in a patient against the cooling tower strain (EY3698)from 1:16 to 1:256 was determined by indirect fluorescent antibody technique, the epidemic of acute febrile illness was concluded as an outbreak of Pontiac fever due to L. pneumophila serogroup 7. The cooling tower was a cylindrical open style, with volumetric flow rate of 130 liter/min, and was used for air- conditioning exclusively to the third floor of the building. The building equipped no air-inlet, and indoor-air of the training room exchanged at every break time through windows of 168 cm in height and 72 cm in width. The cooling tower was not operated for five days before the Group A trainees checked in the Center on 18 August followed by Group B trainees on 19 August. It was speculated that high atmospheric temperature and stagnation of cooling water during this period would lead L. pneumophila to overly multiply, which could be a source of infection by flowing in through opened windows to the training rooms.

Legionellae was found in 57 per cent of 94 cooling towers and 21.8 per cent of 78 other environmental sources. These figures reveal that the existence of legionellae in cooling towers is more prevalent than that found in other environmental sources. Legionella pneumophila serogroup 1 was the most prevalent organism among the legionellae found in cooling towers and other environmental sources. The recovery of legionellae in each province was seasonally independent and was found throughout the year. This is the first report of an environmental survey representing the existence of legionellae in every region of Thailand.

Viet nam is known as an endemic area of melioidosis but its etiologic agent originated in Viet nam was not extensively studied. For the first time, we analyzed the cellular lipid and fatty acid compositions of 15 Vietnamese isolates of Burkholderia pseudomallei, 10 from humans and 5 from the environment. Cellular lipid compositions were analyzed by two-dimensional thin-layer chromatography on silica gel G plates. Cellular fatty acid methyl esters were analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The major lipids in all the isolates were phosphatidylglycerol (PG), two forms of phosphatidylethanolamine (PE-1 and PE-2), and two forms of ornithine-containing lipid (OL-1 and OL-2). PE-1 contained non-hydroxy fatty acids at both sn-1 and -2 positions, while PE-2 possessed 2-hydroxy fatty acids and non-hydroxy fatty acids in a ratio of 1:1. Since snake venom phospholipase A2 digestion of PE-2 liberated 2-hydroxy fatty acids, it was confirmed that these acids are at the sn-2 position of glycerol moiety. In both OL-1 and OL-2, amide-linked fatty acid was 3-hydroxy palmitic acid (3-OH-C16:0), while ester-linked fatty acids were non-hydroxy acids in OL-1 and 2-hydroxy acids in OL-2. The total cellular fatty acid compositions of the test strains were characterized by the presence of 2-hydroxy palmitic (2-OH-C16:0), 2-hydroxy hexadecenoic (2-OH-C16:1), 2-hydroxy octadecenoic (2-OH-C18:1), 2-hydroxy methylene octadecanoic (2-OH-C19CPA), 3-hydroxy myristic (3-OH-C14:0) and 3-hydroxy palmitic (3-OH-C16:0) acids. There were significant differences in the concentration of hexadecenoic (C16:1), methylene hexadecanoic (C17CPA), octadecenoic (C18:1) and methylene octadecanoic (C19CPA) acids among the Vietnamese isolates of B. pseudomallei. However, no significant difference was observed in cellular lipid and fatty acid components between strains of human and environmental origins.

Based on the results of phenotypic characterization, cellular lipid and fatty acid analysis, phylogenetic analysis of 16S rDNA nucleotide sequences and rNA-DNA hybrization, Burkholderia pickettii, Burkholderia solanacearum and Alcaligenes eutrophus are transferred to the new genus Ralstonia, and Ralstonia pickettii (Ralston, Palleroni and Doudoroff 1973) comb. nov., Ralstonia solanacearum (Smith 1896) comb. nov., and R. eutropha (Davis 1969) comb. nov. are proposed. The type species of the new genus is R. pickettii. Type strain of R. pickettii is ATCC 27511T, of R. solanacearum is ATCC 10696T, and of R. eutropha is ATCC 17697T.

The ability of 15 Pseudomonas pseudomallei strains of different origin to survive at 5 degrees C was tested, in comparison with the type strains of Pseudomonas aeruginosa and Pseudomonas cepacia. Viable cells of each strain were suspended in tryptic soy broth (diluted 1:10) at a concentration of about 10(6) CFU/ml. The suspensions were kept at 5 degrees C, and the number of viable cells was determined by counting colonies every 10 days. Four strains of P. pseudomallei, including the type strain for the species, died within 90-100 days at 5 degrees C. Ten other P. pseudomallei strains together with the type strain of P. cepacia survived 160 days or more, although viable counts of these strains decreased to 10(1-2) CFU/ml. The Oklahoma strain of P. pseudomallei and the type strain of P. aeruginosa maintained 10(5) CFU/ml after 170-190 days. It was concluded that resistance of P. pseudomallei to low temperature differs from strain to strain. From the results of our experiments and those reported in the literature of nation-wide soil contamination by P. pseudomallei in France, prejudice regarding the organism as a tropical inhabitant must be corrected.

A 41-year-old Japanese male with uncontrolled diabetes mellitus and alcoholic liver dysfunction developed melioidosis after his business trip to Indonesia and Singapore in 1988. His disease started with spiked fever on the following day after extraction of a tooth, and a liver abscess developed, followed by abscesses in the spleen and in the subphrenic space. In spite of splenectomy and intensive antimicrobial treatments for three months, he developed parotitis, prostatitis, and abscess of the right submandibular gland at 5 to 16-month interval. Pseudomonas pseudomallei was isolated from the blood and pus from each abscess. The lung was not involved. At present, he has returned to work, with continued intravenous instillation of imipenem/cilastatin.

We analyzed the initial and follow-up chest radiographs of 28 patients with culture-positive Legionella pneumonia, and developed a scoring system to quantitate the severity of radiological findings for pneumonia. Intrapulmonary shadows were observed on the initial chest radiograph in 26 patients, but pleural effusion was noted in only one. In one patient the initial chest radiograph had probably been obtained too early to reveal any pulmonary change. Alveolar shadows were noted on the initial radiograph in 21 (81%) patients, and interstitial shadows in 5 (19%). In ten (38%) patients shadows were present in both lung fields. Shadows were prominent in the middle and lower lung fields. A cavity was noted in only one patient, and pleural effusion was also noted at some time during the clinical course in 19 (70%). A large amount of pleural effusion was observed in four patients. The average pneumonia severity score was 3.3 in the 9 patients who survived, and 5.1 in the 17 who died (p > 0.05). The mortality rate was 53% in the 17 patients with pneumonia severity score of 5 or less and 89% in the 9 patients with a score of 6 or more (p > 0.05). Twelve patients died within one week after the initial chest radiograph was obtained. There were no differences among patients with community-acquired infection with or without underlying disease and those with nosocomial infection in characteristic and extension of shadow, presence of pleural effusion, or pneumonia score. The chest radiograph of Legionella pneumonia include bilateral shadow findings characteristic, pleural effusion and rapid progression of shadow, and are clinically useful for diagnosis.

The Working Party for Legionellosis headed by the Japanese Ministry of Health and Welfare processed to standardize the diagnostic procedures for Legionella pneumonia, as the first step to clarify the actual occurrence of patients with this respiratory disease. All the clinical data were collected and analysed on the 28 culture-confirmed patients in Japan during the past 11 years, from 1980 through 1990. The 28 patients were distributed throughout Japan, from Hokkaido to Kyushu. Out of 28 patients, 18 were community-acquired and 10 were nosocomially infected. In 8 of 18 community-acquired patients, any significant underlying disease was not observed. Though it was dominant in the age group in their 60s & 70s, victims were distributed in adults over 20 years of age and even in a new born baby. Only 5 out of 28 patients recovered successfully. From the autopsy findings, in 5 out of the remaining 23 expired patients, Legionella pneumonia seemed to be successfully treated, but other disease or other bacterial pneumonia put an end to the patients. The results of clinical laboratory tests and the efficacy of antibiotics to Legionella pneumonia were essentially the same as those reported in the literature.

Our survey of cooling tower water demonstrated that the highest density of legionellae, >/=10 CFU/100 ml, appeared in water containing protozoa, >/=10 MPN/100 ml, and heterotrophic bacteria, >/=10 CFU/100 ml, at water temperatures between 25 and 35 degrees C. Viable counts of legionellae were detected even in the winter samples, and propagation, up to 10 CFU/100 ml, occurs in summer. The counts of legionellae correlated positively with increases in water temperature, pH, and protozoan counts, but not with heterotrophic bacterial counts. The water temperature of cooling towers may promote increases in the viable counts of legionellae, and certain microbes, e.g., protozoa or some heterotrophic bacteria, may be a factor stimulating the propagation of legionellae.

Based on the morphological, physiological, biochemical and nutritional characteristics, cellular fatty acid and lipid composition, ubiquinone-8 as the major respiratory quinone, guanine-plus-cytosine content of DNA, DNA-DNA homology value, and sequence alignment of 16S rRNA nucleotides, Oklahoma isolate was reidentified as a strain of Pseudomonas pseudomallei.

Based on the 16S rRNA sequences, DNA-DNA homology values, cellular lipid and fatty acid composition, and phenotypic characteristics, a new genus Burkholderia is proposed for the RNA homology group II of genus Pseudomonas. Seven species in this group were transferred to the new genus. Thus seven new combinations, Burkholderia cepacia (Palleroni and Holmes 1981), Burkholderia mallei (Zopf 1885), Burkholderia pseudomallei (Whitmore 1913), Burkholderia caryophylli (Burkholder 1942), Burkholderia gladioli (Severini 1913), Burkholderia pickettii (Ralston et al 1973) and Burkholderia solanacearum (Smith 1896) were proposed.

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-microliters 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 species-proper 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.

The effects of biocidal treatments for cooling towers were examined through the use of chemicals and ultraviolet irradiation to inhibit the growth of legionellae and other microorganisms. In the water of cooling towers without continuous biocidal treatments, heterotrophic bacteria and bacterivorous protozoan first appeared, and then legionellae increased up to 10(4) CFU/100 ml. When a UV sterilizer was connected to the cooling tower, the legionellae count was 1/10 or 1/100 of that in the nontreated tower water. In the water of towers supplemented continuously with the biocidal chemicals, legionellae were not found during a 4-month period. The biocidal treatments tested were proved to suppress the increase of legionellae in cooling-tower water, and thus are useful in preventing the outbreak of legionellosis due to inhalation of contaminated aerosol from the cooling tower system.

Taxonomy is the basic stand point for every field of bacteriology and provides a common language to each worker and researcher in bacteriology throughout the world. Proper identification of isolates from human pathological specimens or from hospital environments is one of the five principal fields of bacterial taxonomy. Before the establishment of classification and stabilization of nomenclature, any isolate can not be identified. Collection of unidentifiable strains is the starting point of new taxonomic investigation. Doctors in medical fields tend to be too much concerned to the infrasubspecific subdivisions, such as serological classification, toxigenicity, resistance to antimicrobics, rather ignoring the classification above the rank of subspecies. Doctors who take care patients and workers in clinical bacteriology laboratories should recognize the importance of bacterial taxonomy in general and use the necessary information in taxonomy selectively.

In view of the wide-spread existence of legionellae in cooling-tower and other environmental water, asymptomatic infection of this organism could occur. In order to verify the possibility of colonization of legionellae at lower respiratory tract of patients with various pulmonary diseases, a total of 22,036 sputum samples from in- and out-patients at National Sanyoso Hospital were examined during a five-year period from September, 1984 to August, 1989. Four (0.073%) out of 5,502 cases were culture-positive for L. pneumophila. L. pneumophila strains were isolated from expectorated, subsequently washed sputum samples of two male and two female patients with respiratory tract diseases. The identification of the isolates was genetically confirmed by the fluorometric microplate DNA-DNA hybridization method. The serogroup (SG) and viable counts of L. pneumophila per ml of sputum of each patient were as follows: 73 y/o female K.H., SG-6, 10(3) CFU; 75 y/o male H.J., SG-5, 10(4) CFU; 61 y/o female M.S., SG-5, 10(5) CFU; and 77 y/o male M.G., not-agglutinable against SG-1-6 antisera, 10(4) CFU. None of the four patients was clinically suspected of legionellosis and antibody titer of paired sera remained 1:64 or lower than 1:32. From these findings, we concluded that L. pneumophila can cause, though quite rarely, asymptomatic infection in human respiratory tract. None of the environmental samples obtained from in- and out-side of the Hospital was culture-positive for legionellae. Thus, the source of infection has remained unknown.

Based on the partial nucleotide sequence analysis of 16S ribosomal ribonucleic acid (rRNA), presence of unique sphingoglycolipids in cellular lipid, and the major type of ubiquinone (Q10), we propose Sphingomonas gen. nov. with the type species Sphingomonas paucimobilis (Holmes et al, 1977) comb. nov. From the homology values of deoxyribonucleic acid-deoxyribonucleic acid hybridization and the phenotypic characteristics, three new species, Sphingomonas parapaucimobilis, Sphingomonas yanoikuyae, Sphingomonas adhaesiva, and one new combination, Sphingomonas capsulata, are described. S. parapaucimobilis JCM 7510 (= GIFU 11387), S. yanoikuyae JCM 7371 (= GIFU 9882), and S. adhaesiva JCM 7370 (= GIFU 11458) are designated as the type strains of the three new species. Emended description of the type strain of S. capsulata is presented.

The type and eight strains of Flavobacterium indologenes were clearly differentiated from the type and two reference strains of Flavobacterium gleum by deoxyribonucleic acid-deoxyribonucleic acid homology data and phenotypic characteristics. Phenotypic characteristics useful to differentiate the two species are presented.

Since the publication of the 8th edition of Bergey's Manual of determinative bacteriology, there has been considerable confusion concerning the names of 2 subspecies and the types of the subspecies of Campylobacter fetus (Smith and Taylor, 1919) Sebald and Véron, 1980. Despite the documentation of the names C. fetus subsp. fetus (Smith and Taylor) Véron and Chatelain and C. fetus subsp. venerealis (Florent) Véron and Chatelain, which appear in the "Approved lists of bacterial names", the influence of Smibert's description in the 8th edition of Bergey's Manual continues. To stabilize the nomenclature and to avoid or reject the use of names which may cause error or confusion, this paper will focus on the legitimacy of the 2 subspecific names of C. fetus proposed by Véron and Chatelain.

An electron microscopic study revealed that, within 30 min after inoculation into the ligated ileal loop of anesthetized mice, cells of Salmonella typhi GIFU 10007 adhered to the M cell surface of Peyer's patch lymphoid follicle epithelium, and induced almost complete destruction of M cells. The M cell cytoplasms were pinched off and extruded from the epithelial lining into the luminal space together with the lymphoid cells primarily enfolded into the corresponding M cells. When two or more M cells were destroyed, a large defect in the epithelial lining was apparent, and a number of bacteria appeared near the basal lamina of the epithelial lining. These findings suggest, as far as anesthetized murine ileal loops and strain 10007 are concerned, that ileal M cells are the target cell at an early stage of S. typhi infection and the infection may further progress to deeper tissues and to the general circulation.