An indirect fluorescent anti-leptospiral antibody technique (FLAi) was studied to detect the antibody in immune rabbit sera. Seven serotypes of leptospira found in Japan were used as the antigens. The fluorescein-labelled antibody used was “Dried Fluorescein-conjugated Goat Antiserum to Rabbit 7S Globulin Laboratory Reagent (Hyland)”. For fixation of the antigen, Kirkpatrik's solution, mixture of ethylalcohol, ether and formalin (6: 3: 1) was found to be fittest. Anti-leptospiral immune sera were prepared by injecting rabbits with 0.1% formalin killed leptospiral suspensions which were made of fresh cultures in Korthof's medium, stored at-20C until use.
The procedure of FLAi was as follows: Leptospiral antigen was smeared on slide glass, fixed, dried, and layered with each of 2-fold serial dilutions of a test serum, sensitized at 37C for an hour in a wet room, and washed several times first with PBS and then with purified water. After drying by the fan, the smear was layered with the fluorescein-labelled antibody, sensitized at 37C for an hour in a wet room following by washing and drying as mentioned above. It was embedded in PBS-glycerin, examined under the fluorescence microscope. The result was evaluated by BV excitation with No.3 filter (Y51).Agglutination was carried out by the Schiifner-Mochtar's microscopic agglutination (MA) method.
Both FLAi and MA gave the highest titers against the homologous serotype antigen respectively, though FLAi showed higher cross reactivity as compared with the case of MA. A rabbit serum immunized with Reiter strain of Treponema produced positive reaction with 3 of 6 antigens at 40-fold serum dilution, while a normal rabbit serum was perfectly negative. In the rabbit infected with a vilulent copenhageni, the antibody was shown by FLAi and MA using the homologous antigen at least for 8 weeks, though FLAi titer tended to lower than MA titer. The reactivity to heterologous antigen was demonstrated by FLAi but not by MA. The mixture of various serotypes may be suitable for epidemiological survey of leptospirosis as the antigen of FLAi.
The detection of leptospira by FLAi technique from urine of pyrogenes-carrying mouse and from blood of copenhageni infected guinea pig was attempted, good stainability of the organisms was demonstrated.
In laboratory possessing the fluorescent antibody technique, it may be possible to detect anti-leptospiral antibody by FLAi supplying no reagents but the leptospira antigen.

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Application of indirect fluorescent anti-leptospiral antibody technique (FLAi) to serodiagnosis of leptospirosis in humans and dogs was studied. Six serotypes of leptospira found in Japan were used as the antigen, they were fixed onto slide glass using Kirkpatrik's solution (Ethanol: ether: formalin=6: 3: 1). The procedure of FLAi was arranged after the FTA-ABS method for serological diagnosis of syphilis. Dried Fluorescein-Conjugated Goat Antiserum to Human IgM (μ-chain specific) Laboratory Reagent (Hyland) and Fluorescein-Labelled Anti-Human γ-Globulin (mainly IgG) Rabbit Serum (Eiken Chemical) were used. Anti-dog γ-Globulin Fluorescent Antibody was prepared by the author. For screening of positive antisera, a mixture of equal amounts of the 6 serotypes was used, the test sera were examined at 5- and 20-fold dilutions.
The antibody titer of the sera from patients of Weil's disease was higher forFLAi than for microscopic agglutination (MA). The sera from 17 undoubted patients at acute stage from whose bloods serotype pyrogenes was isolated, were unexceptionally negative in MA, whereas 9/17 cases to copenhageni antigen and 12/17 cases to the mixed antigen were positive in FLAi (IgM). The serum fromthe convalescent stage showed serotype specificity in MA, whereas considerable cross reactivities inFLAi. There was not so much difference between FLAi (IgM) and FLAi (IgG) titers both in the acute and convalescent sera, nor any consistent tendency between MA and FLAi (IgG) titers in the convalescent sera. From these results, it was considered to be more desirable to use the labelled serum containing both anti-IgM and anti-IgG antibodies for serodiagnosis of human leptospirosis.
In the sera from 4 patients at more than 5 years after the infection, FLAi demonstrated the antibody at the same grade with MA. Out of 5 MA negative sera, one was positive when they were examined at 5-fold dilution, but the others negative. By screening of 295 serum samples from the patients with liver complaints using the mixed antigen of 6 serotypes, 18 were positive at 5-folddilution of the samples and 5 at 20-fold dilution. After absorption with Reiter's antigen, all except 5 turned negative.
FLAi was not always genus-specific in dog, some giving high titer against copenhageni and others against canicola. In the sera of 3 dogs with doubtless infection (1 disease and 2 carrying), MA detected higher titer than FLAi in all cases. It was concluded that the mixed antigen was more adequate than the single serotype antigen for screening purpose in both dogs and human patients.

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Children with pertussis infection or vaccination were studied for the lymphocyte count and the subpopulation in the blood at various stages.
Over the period of the 1st four weeks following pertussis infection, the lymphocyte count was twice as high as that of controls (13.46-13.74 × 10³/cumm). The absolute numbers of T cells and B cells were also greater than that of controls throughout the study period. The number of B cells reached the highest level of 5.43 × 10³/cumm during the first week of illness and decreased gradually thereafter; 4.20 × 10³/cumm at the second week, 3.95 × 10³/cumm at the third week, 3.71 × 10³/cumm at the fourth week and 2.44-103/cumm at the fifth week. T cells continued to increase in number until the fourth week; 7.86 × 10³/cumm at the first week, 9.08 × 10³/cumm at the second week, 9.62 × 10³/cumm at the third week and 9.71 × 10³/cumm at the fourth week and then began to decrease, 7.64 × 10³/cumm at the fifth week. Accordingly T/B ratio showed a steady increase over the first five weeks (1.45, 2.17, 2.43, 2.62, 3.13).
At 0, 4, 15, 22 and 150 hours following the pertussis vaccination, the number of the lymphocytes was 2.91 × 10³/cumm, 4.89 × 10³/cumm, 5.47 × 10^3/cumm, 4.07 × 10³/cumm and 3.84 × 10³/cumm respectively. T cells continued to increase until 15 hours following the vaccination and then began to decrease gradualyi; 2.06 × 10³/cumm before the injection, 3.20 × 10³/cumm after 4 hrs, 4.07 × 10³/cumm after 15 hrs, 3.07 × 10³/cumm after 22 hrs and 2.87 × 10³/cumm after 150 hrs. B cells increased promptly after the vaccination and then gradually decreased; 1.92 × 10³/cumm, 1.83 × 10³/cumm, 1.11 × 10³/cumm, 1.03 × 103/cumm. T/B ratio increased steadily until 150 hours after the vaccination; 1.67, 2.23, 2.76, 2.78. Although B cells and T cells remained elevated throughout the study periods, a remarkable increase in the number of B cells was promptly observed following both natural pertussis infection and the vaccination. It appears likely that the early lymphocytosis reflects an increase in the number of B cells.

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This study was done to assess the incidence and significance of F. meningosepticum in bacteremia and to investigate the clinical backgrounds and bacteriological informations of the patients infected.
In the one year period, 1978, there were 113 patients who had bacteremia in the hospital. From 3 patients F. meningosepticum was isolated singly and from one patient in mixed culture.
Two cases had a coronary artery occlusion, one pyothorax and one severe burn and all of them received antimicrobial agents prior to the isolation.
This study showed that F. meningosepticum is one of the important microorganisms of nosocomial infections.

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A 70-year-old male was admitted with remittent fever and purulent sputum. Chest X-ray revealed infiltrative shadows with multiple irregular cavities in the right lower love. He was treated with ceftezole and tobramycin or sulbenicillin, lincomycin and ceftezole under the diagnosis of pulmonary abscess, but was not improved. Chest X-ray for three weeks after admission revealed a right pleural exudate. w By the puncture of the right pleural cavity purulent exudate was collected and Nocardia asteroides was isolated.
The administration of sulfamethoxazole was immediately started. Clinical symptoms improve completely in about three weeks after the treatment with sulfamethoxazole. It is certain that sulfamethoxazole is very effective for nocardiosis.

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Two patients with acute myelogenous leukemia developed Serratia marcescens septicemia during induction therapy.
One patient developed hemorrhagic cellulitis due to S. marcescens at the peno-scrotal area, which rapidly progressed to fatal septicemia. Both strains isolated from the lesion and blood were non-pigmented and quite resistant to commonly used antibiotics including amikacin (minimum inhibitory concentration, MIC:>100 t, g/ml). They were not agglutinated with any of the commercially available 17 grouping sera. He died in shock within 24 hours of onset of septicemia.
The other patient had severe oral herpetic ulcers preceding the onset of septicemia. S. marcescens was isolated in abundance from the lesion. Both strains isolated from the lesion and blood were pigmented and belonged to serogroup 05. They were sensitive to sulbenicillin and gentamicin (MICs: 25 and 3.13 μg/ml, respectively). She was treated with the combination of these antibiotics in addition to daily granulocyte transfusions, which proved to be effective in association with complete remission of leukemia. No significant rise in serum 0-agglutinin titer was observed.
Serratia appeared in throat and/or urine cultures a few days after initiation of antibiotic therapy for the preceding infection in both cases. Moreover, severe granulocytopenia (<100/mm³) was seen prior to onset of septicemia.
It was suggested that severely immunosuppressed patients receiving broad-spectrum antibiotic therapy may develop Serratia septicemia with increasing frequency in this country.

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