Five strains of Rickettsia tsutsugamushi were isolated in Ehime Prefecture during December 1987 to January 1990. Of these, two strains, the Yamazaki and Noma-3, were isolated at Noma area of Imabari city and three strains, the Kakiwara-l0, -11, -12, at Kakiwara area of Uwajima city. The Yamazaki strain was isolated from a patient of tsutsugamushi disease and the other strains from wild rodents (Apodemus speciosus). These strains showed virulence in euthymic mice. The calculated LD₅₀ of the Yamazaki and Kakiwara-l0 strains showed 10^-3.0 and 10^-1.8, respectively.
The immunofluorescent antibody test using thirty monoclonal antibodies to six representative strains, the Gilliam, Karp, Kato, Irie, Hirano and Shimokoshi, revealed that two strains isolated at Noma area, the Yamazaki and Noma-3, were identified as the Karp type and three strains at Kakiwara area, the Kakiwara-l0, -11, -12, were identified as the Kato type. It was clarified that the serotypic differences were present among the strains isolated in Ehime Prefecture. Moreover, these five strains isolated in Ehime Prefecture did not react with the serotype-specific monoclonal antibodies to the Irie, Hirano and Shimokoshi strains known as the representative strains of so-called new type of tsutsugamushi disease, showing antigenic differences.

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During the 20-year period, 1972-1991, 286 episodes of bacteremia occurred in 200 (45%) of 445 patients with acute leukemia in a hematology ward, giving an incidence of 482 episodes per 1, 000 hospital admissions. The frequency of bacteremia was almost unchanged throughout the study period. The frequency of gram-negative bacilli decreased significantly, however, from 81% of all the isolates for the first half of the study period to 50% for the latter half. Despite the common use of ceftazidime and imipenem during the last 5-year period, Pseudomonas aeruginosa increased in frequency to be the most frequent organism. This was opposite to the decreased frequencies of Escherichia coli, Klebsiella pneumoniae and Enterobacter cloacae. The isolates of P. aeruginosa obtained during this period, all of which proved sensitive to ceftazidime and/or imipenem, were almost equally distributed among five serogroups, although a temporal preponderance of a limited number of serogroups was observed during the preceding 15-year period. On the other hand, the frequency of gram-positive cocci increased from 9% in the first decade to 35% in the latter decade. Staphylococcus epidermidis, Enterococcus species and, to a lesser extent, Staphylococcus aureus were ranked as major pathogens. Among the recent isolates of S. aureus, methicillin-resistant strains virtually replaced methicillin-sensitive ones. Therefore, until more effective ective means for control of P. aeruginosa bacteremia in particular become available, the occurrence of this infection will continue to limit the successful treatment of acute leukemia.

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Picogram quantifies per ml of endotoxin showed a positive limulus test. The concentration more than ten pg/ml of curdlan ((1-3)-β-D-glucan) was also positive to a conventional limulus test, which contains a factor G. One nanogram per ml of CM-curdlan was positive, but higher concentration of 10 pg/ml of CM-curdlan decreased its optical density of the conventional limulus test.
More than one hundred ng/ml of mannan from Saccharomyces cerevisiae did not react with factor G. However, a culture supernatant of Saccharomyces cerevisiae activated factor G. This result suggested that β-glucan activated factor G.
In addition, culture supernatant of Aspergillus fumigatus, Rhodotorula rubra, Candida parapsilosis, Candida tropicalis, Candida krusei also activated factor G, but Cryptococcus neoformans did not activate it.

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We compared the growth of Chlamydia pneumoniae, reference strain TW183 and an isolate from a Japanese infant, AC43 in HeLa229, HL and Hep-2 cells. The mean number of inclusion-forming units was significantly higher on HL cells than HeLa229 cells, when the cells were not pretreated with DEAE-dextran.
When the cells were not pretreated with DEAE-dextran, Hep-2 cells had a higher mean number of inclusion-forming units and a higher yield than other cell lines. Iscove's modified Dulbecco medium enhanced growth of strain TW183 in HeLa229 cells but had a low yield of strain AC43 in Hep-2 cells.

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In 149 patients with purulent meningitis we encountered in the period of 20 years from 1970 to 1990, their clini cal symptoms and prognosis were investigated. Death or sequela was noted in many of the patients having loss of consciousness, stiffness of the members and/or cyanosis as primary clinical symptoms, or having hypoalbuminemia (<2.5 g/dl) and/or thrombocytopenia as abnormal laboratory findings, or having excessive protein levels and/or excessively low sugar levels in cerebro-spinal fluid.
Early initiation of adequate antibiotic therapy, as well as symptomatic treatment using transfusion, steroids and anticonvulsants, are important.

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In a total of 149 children with purulent meningitis we encountered in our institute in the last 20 years, the causatives, the changes in therapeutic management and the prognosis were investigated. The causatives could be detected in 109 patients (73%): H. influenzae; 30 patients (20%), S. pneumoniae; 18 patients (12%), E. coli; 13 patients (9%), GBS; 7 patients (5%) and S. aureus; 6 patients (4%). These ve causatives were detected in 49% of the total patients, or 67% of the patients in whom causatives could be detected. Of these five causatives, E. coli were detected the most frequently in the first half of 1970's, but, in recent years, the detection of GBS, S. pneumoniae and H. influenzae has been remarkably increasing. In spite of progress in antibiotics, the prognosis of the disease due to S. pneumoniae, GBS and S. aureus was poor. In the majority of the patients who died, the death came within five days after hospitalization due to loss of consciousness, convulsion etc. It is therefore necessary not only to initiate strong antibiotic treatment was soon as possible after early diagnosis, but also to take symptomatic measures such as steroidal treatment, treatment of shock etc.

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In this study, role of capsular polysaccharide (CPS) and lipopolysaccharide (LPS) of Klebsiella pneumoniae was investigated in experimental mice pneumonia model.
Inoculation with K. pneumoniae mucoid strain DT-S into mice lung induced expansive, voluminous lethal pneumonia characterized with thickening of the alveolar septa caused by infiltration of inflammatory cell and packing of bacteria within alveolar spaces. On the other hand, mice lung inoculated with K. pneumoniae DT-X, which was non-mucoid mutant isolated from DT-S during natural passage, showed infiltration of inflammatory cell into alveolar spaces but there was no death of mice during the course of this pneumonia. Inoculation of CPS 100p of DT-S strain into mice lung induced lesser extent of accumulation of inflammatory cell than that of LPS 4, ug of this strain. Stimulation of alveolar and pertoneal macrophage with CPS, even at a concentration of 100, ug/ml, induced weaker Interleukin-1 (IL-1) activity than stimulation with LPS 4μg/ml.
These results suggest that since CPS of K. pneumoniae DT-S encapsulate bacteria including LPS, CPS may inhibit chemotaxis of inflammatory cell and IL-1 production of macrophage to be induced by LPS during course of pneumonia. It is speculated that existence of CPS have important role in modulating host response to bacterial LPS, and this effect of CPS may be related with difference of pathological findings of lung and lethality between K. pneumoniae DT-S and DT-X.

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To obtain the basic data on the route of Giardia infection as zoonosis, of many regions in Japan, the feces from 2218 dogs were examined for detection of Giardia cysts. Giardia cysts were detected in 239 of the 2218 dogs (10.9%), which was the same as previous reports from America.None were found from the owners of 51 dogs in which Giardia cysts were detected. The detection rates of each facilities were, 68 of 366 (18.6%) from the breeder's kennels, 169 of 1811 (9.3%) in individual houses, 2 of 42 (4.9%) from research institutes. The detection rate of the breeder's kennels was higher than the other two facilities (p<0.05, p<0.001). The detection rates of Kanagawa and Shizuoka prefectures among 17 regions in Japan were higher than the others (p<0.001, p<0.05). Especially in Shizuoka, the rate of the individual houses was higher than from breeder's kennels. In kanagawa the rates of the individual houses and the breeder's kennels were higher than the mean in Japan (p<0.001). Therefore one must instruct the breeders when teaching health education to included zoonosis, and that the detection rate of the age groups of less than 3 years old was high-221 of 1276 (17.3%).
Since the detection rates of Giardia cysts in the dogs were low, the possibility that human infection acquired from dogs was low. However, some of patients with giardiasis we encountered had never been abroad, and it is not yet clear whether Giardia is strictly host specific or not, so attention should be paid to the possibility of cross-infection between man and animals.

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Since 1989, the number of salmonellosis cases caused by S. Enteritidis has increased considerably in Japan.
Genetic differentiation of 385 strains isolated from January 1982 to December 1988 and January 1989 to April 1991 were used for plasmid profiles.
Plasmids were found in 377 out of 385 strains; therefore, only 8 strains carried no plasmid. Among 377 strains, 15 different plasmid profile types (OP-1 to OP-15) were classified.
The most common plasmid profile types from 1982 to 1988 were OP-7 (70 kbp) and OP-8 (70 kbp and 2 kbp). On the other hand, the most common plasmid profile types from 1989 to 1991 were OP-1 (60 kbp) and OP-2 (60 kbp and 54 kbp).
Serovar-specific virulence 60 kbp plasmids of S. Enteritidis were identified in 7 plasmid profile types (A total of 200 strains). In the other plasmid profile types, 70 kbp plasmids were found in 5 plasmid profile types (A total of 173 strains). In restriction enzyme analysis of 70 kbp plasmid DNAs obtained from 5 plasmid profil types of S. Enteritidis, we found that these plasmid DNAs shared both 60 kbp and 10 kbp fragments. These results indicate that these plasmid profile types also carried serovar-specific virulence plasmids of S. Enteritidis.
The strains of plasmid profile type OP-2 were SA (sulfisoxazole) and SM (streptomycin) resistance, and the 54 kbp plasmids in the strains of OP-2 were transferred by bacterial conjugation into the E. coli strains. All transconjugants acquired SM resistance.

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Antibodies against rubella virus in human sera were measured by ELISA and antibody titers werecalculated by the parallel line assay method.
The dose response regression curves of standard sera and test sera containing IgG antibodycalculated by the parallel line assay method showed linearity, and were parallel to one another.However, the regression lines of sera positive for IgM antibody against rubella virus were not parallelto one another in the low dilution region, but parallel in the higher dilution region.
A good correlation was observed between the rubella IgG antibody titers measured by thehemagglutination-inhibition (HI) test and those calculated by the parallel line assay method. Thecoefficient of the correlation was 0.781.
Time-course studies of IgG or IgM antibody titers against rubella virus in rubella patients or invaccines with MMR vaccine indicated that the ELISA was more precise and specific method than theHI test.
Thus, the parallel line assay method using ELISA is considered to be a more useful method for thedetection and quantification of antibodies to rubella than the conventional HI test.

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We evaluated the usefulness of a Nested PCR method for detecting Legionella pneumophila. This method resulted in L. pneumophila specific detection as far as we evaluated. The first and second step PCR achieved the sensitivity as small as 10 pg and 10 fg of the target DNA, respectively. In the detection from Legionella seeded sputa, the method could detect 0.1 cfu/ml of the bacteria, and it took about 12 hours to detect the target DNA. We demonstrated that the Nested PCR method was superior in sensitivity and rapidity for isolation of the bacteria to the conventional using low pH treatment and selective media for Legionella.

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Aerobic and anaerobic cultures as well as a Gram stain and wet mount preparations were made of vaginal swabs in twenty patients with clinical bacterial vaginosis. Mobiluncus spp. were detected in 7 cases (35%). Cultures appeared to indicate that mixed abnormal flora between aerobic and anaerobic bacteria are found in bacterial vaginosis, and that Mobiluncus spp. may play a role in bacterial vaginosis.

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Non-radioisotopic, alkaline phosphatase-labeled DNA (AP-DNA) probe tests for the identification of Mycobacterium tuberculosis complex (Mtb) and Mycobacterium avium complex (MAC) were evaluated. The overall agreement, sensitivity and specificity of the AP-DNA probes for Mtb and MAC were 100% respectively compared with the conventional biochemical method. Because the procedure is rapid (it can be completed approximately 120 min), safe (it does not use radioisotopes) and convenient (it does not need special the equipment to be performed), it can be easily performed in any clinical laboratory.

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The effectiveness of long-term chemotherapy using Erythromycin (EM) for chronic lower airway infection has been practically proved. However, there still exist some ineffective cases, or cases in which the clinical effect was scarcely seen. In the present study was administered Clarithromycin (CAM) to such cases and found that CAM was effective in alleviating symptoms in some of them. The results were presented along with clinical findings and other basic studies. The subjects were 4 cases in which EM was either ineffective or low in its clinical effect. The subjects consisted of 1 case of DPB and 3 cases of bronchiectasis. EM was clinically ineffective in 2 cases and slightly effective in the two others. The pathogen was Pseudomonas aeruginosa in all cases. The dosage of EM was 200-1200mg/day. The period of administration ranged from 2 years to 6 years 9 months. CAM was given orally after meals at a dose of either 200 or 400 mg/day. Chemotherapy had continued for 3-8 months at the time of final observation (Feb. 1992). Clinical effectiveness was evaluated on the basis of sputum volume and PaO₂ examination, as well as evaluation by the patients themselves. As a result, in all 4 cases, reduction in sputum volume and improvement of PaO₂ were observed. All subjects evaluated CAM therapy as being more effective than EM therapy. Moreover, it was found that CAM inhibited both the elastase and leucocidin produced in one of the ineffective cases by P. aeruginosa, whereas EM didnt. In studies on cytokines (IL-2, IL-4, IFN-γ) in mice, levels elevated in the mice given EM and CAM for 28 days. Moreover, in a study on the changes of IL-2 with the passage of time, IL-2 elevated earlier in the mice given CAM. On the basis of these findings, it was suggested that CAM had effect in EM-ineffective cases of chronic lower airway infection.

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Relationship between the normal throat flora and pathogenic bacteria recovered from the throat in 139 children with upper respiratory tract infections in winter were studied using quantitative analyses. Pathogenic bacteria examined include S. pyogenes, H. influenzae, S. aureus, and S. pneumoniae, and the normal floras include α-streptococci, γ-streptococci, Neisseria species, and Micrococci. Children with S. pyogenes in their throats (S. pyogenes group) were examined with antistreptococcal antibodies such as anti-streptolysin 0, anti-streptokinase, and anti-deoxyribonuclease B.
Eighty seven pathogenic bacteria were recovered from 72 children (51.8%) out of 139. S. pyogenes and S. pneumoniae groups showed significantly lower α-streptococci and γ-streptococci in incidence of appearance when compared with children with the no pathogenic bacteria in their throats (no bacteria group). H. influenzae group showed significantly lower γ-streptococci and higher Neisseria sp. in incidence of appearance compared with the no bacteria group.
Positive cases for anti-streptococcal antibodies showed a significantly lower α-streptococci in number compared with negative cases for antibodies and the no bacteria group, and a significantly lower γ-streptococci in incidence of appearance compared with the no bacteria group.
These data suggest that the normal throat flora may have a role in prevention of colonization by the pathogenic bacteria in vitro, as were shown in vitro by many authors, and that the quantitative analysis of the normal flora is useful because this methodology might reveal whether the bacteria recovered from the throat show the pathogenicity.

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Miconazole (400.1200 mg/day) was administered to patients with deep mycosis and suspected deep mycosis, and the efficacy evaluated. 13-Glucan was determined as the early diagnostic parameter of deep mycosis, and the relationships between the clinical efficacy of miconazole and the titers of β-glucan were also evaluated.
Forty-nine cases were evaluated, including 2 cases of deep mycosis and 47 cases of suspected deep mycosis. Most of the patients had hematological malignancies.
The rate of efficacy was 100%(2/2) in deep mycosis, 66.0%(31/47) in suspected deep mycosis, and 67.3%(33/49) in total.
β-Glucan was determined in 39 cases before the administration of miconazole. The rate of β-glucan positivity was 100%(1/1) in deep mycosis and 44.7%(17/38) in suspected deep mycosis.β-Glucan was also determined before and after the administration of miconazole in 11 cases. The titers of β-glucan became negative in 6 cases, decreased in 2 cases and increased in 3 cases. Thus, the β-glucan titers became negative or decreased in 72.7%(8/11) of the cases. Efficacy of miconazole was 83.3%(5/6) in the cases in which fi-glucan became negative, 50.0%(1/2) in the cases in which the titers of β-glucan decreased, and 33.3%(1/3) in the cases in which the titers of β-glucan increased.
Miconazole was effective in the treatment of deep mycosis, and the titers of β-glucan correlated well with the clinical efficacy icacy of miconazole. The determination of pglucan appears to be useful for the diagnosis of deep mycosis.

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A 59-year-old male was admitted to our hospital in Jan. 1991 with complaints of general malaise and palpitation. Laboratory findings on admission showed anemia, thrombocytopenia and leukopenia consisted of 2.0% myeloblasts with Auerbodies. The bone marrow study showed granuloid hyperplasia with 45.5% myeloblasts. The diagnosis of acute myeloblastic leukemia (M1) was made. After BHACAMP therapy, he obtained complete remission. However, he complained of fever and cough, and his chest X ray film showed a focal infiltrative shadow in the right upper lung field. Antibiotics for bacteria and fungus were administrated and the abnormal shadow improved in a week. However, as he had hemosputum, the bronchoscopic examination was performed, and multiple ulcers covered by yellow-white tissue were revealed on the wall of the trachea and bilateral main bronchi. Biopsy specimens obtained by transbronchial biopsy showed bronchial aspergillosis. Though intravenous infusion and inhalation of amphotericin B were effective for aspergillosis, he had a relapse of the leukemia and died in autumn, 1991.

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Continuous ambulatory peritoneal dialysis (CAPD) was introduced to Japan ten years ago and was established as the treatment for end-stage renal disease along with HD. Although the incidence of peritonitis in CAPD has decreased by educating the patients and parents and the improvement of various devises of CAPD, peritonitis is still one of the major complications of CAPD. Fungus is a rare pathogen for peritonitis in CAPD, but it must be considered as a causative agent in cases of intractable peritonitis.
This report describes the first case of Trichosporon beigelii (T. beigelit) peritonitis in CAPD in Japan. A nine year old boy with chronic renal failure due to bilateral vesicoureteral reflux was given CAPD treatment four years prior to admission. This patient had been admitted to our hospital frequently because of recurrent bacterial peritonitis. The peritonitis in CAPD was usually treated by changing the peritoneal fluid and antibiotic treatment. In this case T. beigelii was proved to be a pathogen of peritonitis by culture of CAPD fluid and also serum antibody titers. T. beigelii infection was successfully eradicate from the peritoneal cavity by administration of MCZ and by the removal of peritoneal catheter. The patient was switched from CAPD to HD. In the case of intractable peritonitis in CAPD, rare fungal pathogens such as T. beigelii must be considered as a causative agent.

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