The inhibitory effect of Clostridium butyricum MIYAIRI 588 against various enteropathogens was investigated in mixed cultures. It was observed that C. butyricum M588 inhibited the growth of Vibrio cholerae O1, V. cholerae non-O1, Aeromonas hydrophila, and Shigella flexneri. Considering that the interaction between C. butyricum and Shigella is especially important because of their proliferation site in the lower intestine, further examinations were carried out on Shigella in particular. Results were as follows:
1) In BHI broth culture of Shigella, the pH of culture fluid went down to 5.2, but the growth of Shigella was not inhibited.
2) In the mixed culture of Shigella and C. butyricum, the growth of Shigella was inhibited, nevertheless the pH of the culture fluid was 5.6.
3) In the mixed culture with phosphate buffered BHI maintaining the pH higher than 6.0, the growth of Shigella was inhibited.
4) In case of pure culture of C. butyricum in BHI broth, the pH of culture fluid indicated 5.5, and Shigella failed to grow in the cell free culture supernatant.
5) The growth of Shigella was not inhibited in the culture supernatant when the pH was adjusted at 7.2.
These results suggested that the inhibition of Shigella in the mixed culture with C. butyricum was not due to a single factor such as pH or fatty acid etc. but due to multif actors including live cells of C. butyricum.

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The usefullness of persistently infected C6/36 (C6/J-121) cells with Japanese encephalitis virus (JEV) for a rapid serodiagnostic test was examined with the sera of men, swine and laboratory animals by indirect immunofluorescent antibody (IFA) technique.
Detection of specific antibodies was completed within 3 to 5 hours.
Nonspesific fluorescence frequently observed in other IFA tests was few.
The sensitivity of antibody detection and the serological specificity of IFA (IgG) were similar to those of hemagglutination-inhibition (HI) test.
For the detection of IgM antibodies in sera of JE-patients and naturally JEV infected swine, this method was found to be simpler and more sensitive and rapid than the conventional HI test which required 2-ME treatment of the sera.

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Mycobacteria other than Mycobacterium tuberculosis isolated in Tokyo University Hospital during 1970-1980 were identified to the level of species using a commercial DNA probe Kit (Gen-Probe Rapid Diagnostic System for Mycobacterium avium Complex; Gen-Probe Corp., California, USA). The cell count to give a positive result, that is, the absolute hybridization of more than 10%, was 5×10⁶ CFU/ml for M. avim and 1×10⁶ CFU/ml for M. intracellulare, respectively. At this condition, compared with the conventional biochemical test, the DNA prove procedure showed sensitivity of 98.8% and specificity 100%. The agreement of the identification between the two methods was 99.1%. The isolation frequency at the level of species among 104 isolates tested in this method was: M. avium72.1%, M. kansasii 7.7%, M. intracellulare 3.8%, M. gordonae 3.8% and others 12.5%. Thus, this DNA probe test to identify and distinguish M. avium complex gave a satisfactory result in its simplicity and accuracy.

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Bovine colostrum whey and immunoglobulins were prepared. Their characteristics and anti-viral activities were studied: IgG, IgA and IgM were found in bovine colostrum. Most IgG was polymerized. Although neutralization activities against bovine, simian and human rotaviruses existed, anti-human adenovirus antibody was not found.
Effects on prophylaxis and treatment for rotavirus gastroenteritis were expected.

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The Plasmid DNA patterns of 22 penicillinase-producing Neisseria gonorrhoeae (PPNG) strains isolated from male patients with genococcal urethritis in 1988 at Tokyo Metropolitan Taito Hospital were determined.
The 3.3 megadalton plasmid, the so called the Africantype plasmid, was present in 3 isolates out of 22 PPNGs, all of which occurred with a conjugative 24.5 megadalton plasmid. 4.4 megadalton plasmid was present in 19 isolates out of 22 PPNGs, 5 of which occurred with a conjugative 24.5 megadalton plasmid.
The infective sources of the 3 cases of the African type PPNGs were found as 1 from philippines and 2 from prostitutes, so called Soap Land Girl in Japan. And they suggests that the African type PPNGs are already common in Japan.

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C. trachomatis isolation by culture method using McCoy cells and direct immunofluorescence test (DFA) were carried out on specimens collected from patients with suspected chlamydial infections and the results were compared.
Follow-up survey was also made on C. trachomatis positive patients. The results are summarized as follows:
1) One thousand and eighty-six specimens were examined. Postitive-coincidence ratio obtained from both methods was 80% and negative-coincidence ratio was 92%. The antigen detection by the DFA exceeded a little bit in positive ratio than that by the culture method.
2) C. trachomatis was positively detected in 31 of 94 men (33%) and 5 of 27 (17%) women during periods from 7 days to 2.5 years in follow-up examinations.

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The basic studies on detection and titration of the 60-kDa cross-reacting protein antigen (CRPA), which is common to gram-negative rods, by reversed passive gelatin agglutination test with monoclonal and polyclonal antibodies were performed. The gelatin particles sensitized with 500μg/ml of the monoclonal antibody had an ability to react with 200 ng/ml of the purified CRPA for 1.5 h.
In contrast, the particles sensitized with 200μg/ml of the polyclonal antibody had an ability to react with 25 ng/ml of the purified CRPA for 1.5 h.
In addition, the detection of the 60-kDa CRPA in urinary tract pathogens, which consisted of 60 bacterial strains representing 14 species, was carried out by using the gelatin particles sensitized with 200μg/ml of the polyclonal antibody.
The CRPA was detected in all gram-negative rods of urinary tract pathogens, but not in grampositive cocci.
These reuslts suggested that the detection of the 60-kDa CRPA by reversed passive gelatin agglutination test was a rapid and simple method for screening gram-negative bacteriuria.

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To obtain basic data on the route of Giardia infection as zoonosis, we examined feces from 80 dogs and 16 cats for Giardia cysts and evaluated the detection rates. In addition, familial infection was studied in 3 family members of the patient with giardiasis.
Giardia cysts were detected in 10 of the 80 dogs (12.5%) but none of the cats. Of the 10 dogs from which Giardia cysts were isolated, 6 had been maintained by the breeders. Tow other dogs were for examination in the research institutions. None of the family members of the patient with giardiasis had this infection.
Since the detection rates of Giardia cysts in the dogs and cats were low, the possibility that human infection is acquired from dogs or cats seemed to be low. However, some of patients with giardiasis we encountered had never been abroad. This fact together with the presence of Giardia cysts in the dogs despite the low detection rates suggest that attention should be paid to the possible association between human giardiasis and pets since the number of people keeping pets is increasing.

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This study showed several accumulated data through ten years from our experiencein hematopoietic disorders and associated infections, which has been analyzed by the Hanshin Study Group of Hematopoietic Disorders and Infections.
Since 1979 to 1988, our group had evaluated the sorts of causative organisms andthe efficacy of various antibiotics therapy in 2119 cases of infectious diseases associated withhematopoietic disorders. On behalf of evaluating the changes of disease profile for ten years, we divided the accumulated data into three phases; former phase the first three years, middle phase the second three years and late phase the last four years.
There was no significant difference in the frequency of various hematopoietic disorders among the three phases. Each leukemia patients occupied 77% of all cases. Sepsis suspectedis the most frequent infectious disease accounting for 68.8%. The other infectious diseses were 8.4% of the sepsis, 14.8% of the respiratory infections and 3.1% of the urinary tract infections. Comparing the frequency of infections among the three phases, the respiratory and urinary tract infections inclined to decrease.
Of the 532 strains isolated from 2119 cases and identified as causative organisms, gram-negative bacilli occupied 62.8% and gram-positive bacteria 36.5%. In comparing the percentage of gram-negative bacilli among the three phases, it showed a decreasing tendency in order former phase 63.6%, middle phase 76.4% and late phase 43.8%. Pseudomonas, however, had been isolated at almost constant ratio through ten years. On the other hand, the ratio of gram-positive bacteria isolated were 34.5% in former phase, 23.6% in middle phase and 56.3% in late phase, showing increasing a tendency through the period.
Twenty-three kinds of antibiotics were administered by intravenous drip infusion. The efficacyicacy rate was 43.9% to 67.2%, In paticular, effectiveness of antibiotic therapy oftendepends on the change of peripheral neutrophil counts from the onset and during the therapy. The efficacyrate, however, was 36% even neutrophil counts have not shown the tendency of increase from less than 100/μl.

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In this study of Chlamydia trachomatis (C. trachomatis) from genital origin in McCoy cells, minocycline (MINO) was added to infected cell cultures at specific stages of their growth cycle. Sequential observations were made at each stage by electron microscope with the following results:
1) The minimal inhibitory concentration of MINO against C. trachomatis in McCoy cells was 0.025μg/ml.
2) When MINO and chlamydial inoculation were simultaneously added (inoculum containing 1μg of MINO per ml), MINO did not inhibit attachment and invasion of elementary bodies (EBs) to McCoy cells in both cases of inoculation, wiht and without centrifugation.
As 48 hours after inoculation 74% of the infectious EBs in McCoy cells were converted to early intermediate forms (IFs). This suggests that MINO did not inhibit the transformation of EBs to early IFs. Under the influence of MINO, even after 48 hours, reticulate bodies (RBs) and chlamydial inclusions were not recognized. At the stages of transformation of infectious EBs to early IFs, duplication of the cell wall and electron lucent areas in the cytoplasm were observed.
3) Meanwhile, in the case of replacing the media with one containing MINO (1μml) at 12 hours after inoculation, the conversion of RBs to late IFs was blocked.
4) Electron microscopically, the average numbers of late IFs and EBs per cross section of the inclusion at 24 hours after inoculation, were 0.64 and 0.17, respectively. At 48 hours after inoculation without MINO (control), the average numbers of late IFs and EBs were 10.5 and 65.6, respectively. When MINO (1μg/ml) was added at 24 hours after inoculation and observed at 48 hours, the average numbers of late IFs and EBs numbered 0.10 and 0.67, respectively. This indicates A) statistically, a significant decrease (0.64→0.10) for late IFs and a significant increase (0.17→0.67) for EBs compared with the average number of those at 24 hours after inoculation, B) when compared with the average number of those at 24 hours after inoculation, the sum of late IFs and EBs were nearly the same (0.64+0.17=0.10+0.67), C) somewhat similar ratios of late IFs to EBs (10.5: 65.6=1: 6.2=0.10: 0.67=1: 6.7) compared to the average number of those at 48 hours after inoculation without MINO (control).
B) suggests that the conversion of RBs to late IFs was blocked by MINO.
A), B) and C) suggest that the conversion of late IFs to EBs was not inhibited by MINO.
As MINO has been identified as an inhibitor of protein synthesis in bacterial ribosome, new protein synthesis may not be necessary for EBs to invade McCoy cells and for infectious EBs to convert to early IFs. New protein synthesis may be necessary or early IFs to become RBs and for RBs to convert to late IFs. It also may not be necessary for late IFs to transform to EBs.

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Serological typing of group B streptococci is important for the epidemiological study of group B streptococcal infections. These days, non typable (NT) strains by conventional serotypes were on an increase.
In 1984, strain “M9” was isolated from a pregnant woman in Meijo Hospital, Nagoya City; herein the antigenicity and prevalence of strains typed as “M9” were investigated and discussed.
The results obtained were as follows:
1) It was confirmed that strain “M9” had a new polysaccharide antigen, different from conventional types, Ia, Ib, II, III, IV and V, type candidate NT6 and type candidate 7271, as found by precipitation and precipitation absorption reaction. This procedure, moreover, was useful for differentiation type candidate NT6 and 7271 from “M9” because of their provisionality.
2) Group B streptococci typed as “M9” were isolated not only from carriers but from patients who were newborn babies and suffered from sepsis.
3) Strain “M9” was not necessarily located in Nagoya City but in Chiba and in Kyoto, this type was isolated from clinical materials already in 1979.
4) It was observed that polysaccharide “M9” liked to combine with protein “R” and without other proteins, as our collection extends.

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Eight cases of diphyllobothriasis have been experienced in the Juntendo University Hospital.
Seven of the 8 patients excreted tapeworm fragments. Eggs of Diphyllobothrium latum were found in the feces in 5 cases. One patient had a history of ingestion of raw trout (Sushi), and 2 raw salmon. One might have been infected in foreign countries, and 3 could not tell the source of infection.
Bithoinol was administered orally to 7 patients. Four of the 7 excreted the worm and the scolex was recognized in three of the four. Neither recurrence nor abnormal findings have been recognized so far.

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We analysed retropspectively 48 hospitalized patients with fever of unknown origin (FUO) from 1982 through 1988.
The criteria of FUO were (1) temperature of more than 38.3° documented on several occasions (2) overall duration of illness more than three weeks, (3) uncertain diagnosis till one week after hospitalization.
Of this group of FUO, 25 patients (52.1%) were found to have infections, 8 patients (16.7%) had collagen disorders, 7 patients (14.6%) had neoplastic disorders, 3 patients (6.3%) were crohn disease and 5 patients (10.4%) were undiagnosed.
Among infectious diseases, chronic tonsillitis was the most frequent (5 patients: 20%) and they were diagnosed by the provocative examination. Non bacterial meningitis and cervical lymphadenitis were diagnosed in all 3 patients (12% in all), Adult Still's disease was found in 3 patients (37.5%) and systemic lupus erythematosus (SLE) in 2 patients (25%) in collagen disease. Immunoblastic lymphadenopathy was diagnosed in 3 patients (42.9%) of malignant diseases. Three cases of Crohn disease were revealed in all the patients of the miscellaneous group.
Duration of fever was relatively short in infection diseases compared to malignant and Crohn diseases. The most common laboratory abnormality is an elevated erythrocyte sedimentation rate (89.6%).
As the final diagnosis of FUO are changing with the development of diagnostic tehniques, a new criteria of FUO is necessary.

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A 46-year-old man was admitted to our hospital on Mar. 16, 1988 with the chief complaint of productive cough. The chest roentgenogram and tomogram showed a tumorous shadow in the right upper lobe, accompanied with stenosis of the trachea and the right main bronchus. Bronchofiberscopical examination confirmed a nodular tumor protruding into the lower part of the trachea from the right lateral wall and nearly complete obstruction of the right main bronchus. Pathological specimen obtained by transbronchial biopsy revealed “low differentiated adenocarcinoma”. It was highly suspected that the primary lung cancer had directly invaded the trachea and the right main bronchus. His symptoms and roentgenological findings remarkably improved after radiation therapy. He was discharged on May 12.
On Sep. 14, he was admitted to our hospital again because of hoarseness, general fatigue and increasing dyspnea. The chest CT demonstrated severe stenosis of the trachea, which was treated with another radiation therapy. Although his symptoms diminished, he had a sudden onset of high fever on Oct. 15. Immediately a blood culture and transtracheal aspiration (TTA) were performed. Gram-negative bacilli were isolated from the blood culture four days later. The administration of fosfomycin and tobramycin was started. However he died because of massive hemoptysis on Oct. 23.
Several days after the death, an isolated strain was identified as Capnocytophaga ochracea by the biochemical characteristics. Culture of sputum obtained by TTA was negative for Capnocytophaga ochracea.
Septicemia due to Capnocytophaga spp. is very rare and only one case (due to Capnocytophaga sputigena) has been reported until now in Japan. Our patient is thought to be the first case of septicemia due to Capnocytophaga ochracea in Japan.

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