Waste of seawater used for cooling water for machines in some kinds of industries in a littoral district in Sakai City, comes under the application of BOD control of the pollution control ordinance in Osaka Prefecture. But unfortunately the BOD measurement method of seawater sample has not been established.
In order to establish the measurement method, we studied the strength of activation of aerobic bacteria in the incubation water used and the dilution water.
From our results it was proved that we can use the incubation seawater consisting of more than 10³ cells/ml general bacteria count and more than 0.50 μg/l ATP (Adenosin triphosphate) as the incubation water and the artificial seawater as the dilution water.

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Isepamicin (ISP) and piperacillin (PIPC) were shifted to the urinary concentration by employing an in vitro complicated cystitis model operated by a computer-controlled automatic simulator for urinary concentration, and administrated to bacteria in the urinary bladder model (Pseudomonas aeruginosa: P. aeruginosa, initial cell concentration: 10⁷ cfu/ml). In this case, effects by single treatment with ISP or PIPC on cell number curves were examined. Further, significance of thecombined treatment with ISP and PIPC were investigated by changing the order of each treatment. And following the results were obtained.
1. In a single treatment with PIPC the cell concentration was minimum (104 cfu/ml) at 9th hour after its treatment and thereafter, regrowth to the same level as the initial concentration was obserbed at 16th hour.
2. In the case of single treatment with ISP, the cell concentration became minimum (10² cfu/ml) at 13th hour after the treatment and raised to the same concentration as the initial one at 25th hour.
3. In the combined treatment, the cell concentration was minimum (less than 10¹ cfu/ml) at 26th hour in the case of prior treatment with ISP. Thereafter, regrowth was observed and the cell concentration at 42th hour reached to the initial cell concentration.
4. In simultaneous treatment with ISP and PIPC, the cell concentration at 24th hour was minimum (10¹ cfu/ml) and reached to the same level as the initial one after regrowth.
From these results, it was found that the combined treatment with ISP and PIPC caused more reduction of the cell concentration than either single treatment. Further, regrowth of the cells was suppressed for longer duration. As to the order of those treatments, it was found that the simultaneous treatment with ISP and PIPC was the most superior for the rate of reduction in the cell concentration and prior treatment with ISP was most superior to reduce the cell concentration. Conversely, prior treatment with PIPC was the most superior in respect to the duration when regrowth of the cell was suppressed.

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In Okinawa prefecture, prevalence of hepatitis B surface antigen (HBsAg) among blood donors is 3.5% and is twice as high as the average for the whole of Japan (1.5%), and is the highest in Japan (p<0.005). In contrast, mortality rates of both liver cirrhosis (LC) and primaryliver cancer (PLC) in Okinawa are the lowest in Japan. Many epidemiological studies have shown that the positive rate of HBsAg correlates with mortality rate of PLC. To elucidate the cause of this epidemiological discrepancy, cross-sectional seroepidemiological studies and a prospective clinical study were conducted.
In the cross-sectional studies, the following results were obtained;
(1) Positive rate of HBsAg among patients with LC in Okinawa was 15.2% and lowerthan the average for the whole of Japan (23.4%). A similar comparison among patients withhepatocellular carcinoma showed 24.4% in Okinawa Vs. 31.4% in the whole of Japan. (2) The age-specific hepatitis Be antigen positive rate among 829 HBsAg positive health examinees tend to decreasewith increase in age; 50% in<20 years old age group, 15.7% in third decade and 2-3% or less in 30 or more age group. Of the 829, 431 HBsAg positive subjects were referred our liver out-patient clinic.Then, of the 431, 27 (6.3%) were diagnosed or suspected as having chronic hepatitis and one (0.2%) was diagnosed as havingcirrhosis. Of the 431, 381 (88.4%) were diagnosed as healthy HBsAg carrier, the great majority (94.0%) of whom had positive reaction of and-HBe antibody and normal values of both GOT and GPT.
On the other hand, the prospective clinical study showed that; Among 24 initially HBeAg positive patients with chronic type B hepatitis, all of whom were followed-up for more than 12 months, 14 cleared HBeAg and subsequently 10 of the 14 acquired anti-HBe antibody. The cumulative clearance rate of HBeAg within the first two years was 56.3% and the annual clearance rateof HBeAg was 25.6%.
The tendency of early disappearance of HBeAg among HBsAg carriers during their life time or the course of chronic hepatitis might lead to the low death rate of hepatocellular carcinoma, particular that of HCC associated with hepatitis B virus infection in Okinawa.

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Before 1983, S. hadar was seldom isolated from man, animals, food or the environment in Japan: only one strain having been isolated from man and one from the environment. In subsequent years there has been a progressive increase in the number of isolations. S. hadar is now one of the commonest serotypes isolated from cases of sporadic diarrhea in Shizuoka Prefecture (Table 1). However, the epidemiology of S. hadar is not clearly understood. Reports on the clinical features of S. hadar gastroenteritis are also scarce. We examined the clinical symptoms of 15 cases of S. hadar gastroenteritis. S. hadar was encountered in patients of all ages. Infants and young children below 10years of age constituted 47% of all cases. Seventy per cent of young children below 10 years of age experienced fever of more than 39°C. This incidence was significantly higher than that of general Salmonella gastroenteritis observed in our previous study, in which fever of more than 38°C was noted in 61% of the children. We found one case of presumptive person-to-person spread. No other household contacts of index patients suffered from diarrhea during the same period. Systematic examination for Salmonella contamination was performed for poultry farms, broiler chickens, broiler processing plants and meat on the market. 259 Salmonella strains were isolated from 1197 samples. S. hadar accounted for 37.1%(96) of all isolations (259). A drug resistance test was performed for 51 strains of the diarrhea cases and 67 strains of the environment. The pattern of the distribution of MICs of 9 drugs was similar in the two groups. Two clear peaks of resistant and sensitive strains to ABPC, KM and CER were noted for both groups. The proportion of resistant strains was higher in the case of strains from humans (47/51, 92%) compared to strains from the environment (35/67, 52%). Phage type determination was performed for 30 strains of S. hadar (Table 7). Phage type 2 comprised 50%(15 strains), followed by types 11 and 22 (17%, 5 strains each). The dominant occurrence of type2 among strains isolated from sporadic diarrhea patients or poultry meat on the market seemed to indicate that such meat is an important source of infection in humans. The plasmid profile for 51 strains from sporadic diarrhea patients and 67 strains from the environment is shown in Table 8. The wide distribution of the plasmid profile in both groups made the value of this examination as an epidemiological marker rather poor. However, the similarity noted in the strains collected on the same day or in the same place showed its value in limited situations.

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A systematic examination was performed for environmental food contamination by Salmonella in poultry farms, broiler chickens, broiler processing plants and meat on the market. Salmonella was isolated from 219 of 1197 samples and the serotypes showed a wide distribution. (Table 1-1). S. hadar accounted for 37.1%(96) of all isolations (259). The contamination rate of broiler chickens on arrival at broiler processing plants is relatively low. However, in the broiler processing plants, containers, processing machinery, cooling water and slaughters were highly contaminated by Salmonella, S. hadar being the most prominent serotype in the plants. 64% of chicken meat on the market was contaminated by Salmonella, S. hadar being the second most prominent serotype. 11% of the pork and none of the beef or horseflesh was contaminated by Salmonella. These results indicate that poultry is the main source of S. hadar infection in humans. However, no S. hadar was isolated from cultures of 119 samples of feed for chickens from each delivery (Table 1-1). Thus, as the main source of infection by S. hadar of broiler chickens, an association with the feed seems to be ruled out. S. hadar was isolated at three of 18 poultry farms within Shizuoka Prefecture.
Follow-up studies were performed at the three poultry farms which revealed that in two of them, Salmonella was completely eradicated on completion of disinfection. In the other one farm, which is still being disinfected, various strains of Salmonella including S. hadar still survived. We conclude that the main cause of the problem is the magnification of contamination of Salmonella-free material during the process at the broiler processing plants. Frequent washing and disinfection at broiler processing plants will reduce the magnificantion of contamination. The control and prevention of S. hadar infections in poultry farms are also important. The possibility of the eradication of Salmonella was confirmed by the complete disinfection of the contaminated chicken houses by means of a defined method after removing the chickens. Further attempts to eradicate the organism from commercial machinery should be made.

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Modified microdilution plate hybridization was used for genetic identification of 25 Legionella species.
The cell wall of the isolate was broken down by glass beads. And the DNA was extracted, labeled photo-reactive biotin, and hybridized with reference DNAs immobilized in microdilution wells. Hybridized DNAs were detected by colormetric method.
Each type strain of 25 Legionella species was clearly differentiated by this method.
Among 103 clinical and environmental Legionella strains, 97 strains were genetically identified by this method. 60 strains were identified as the same species as by the conventional method. Concerned with ten strains that had been serologically identified as L. bozemanii, two strains isolated from the human lung were genetically identified as L. bozemanii, but the remaining environmental 8 strains were identified as L. anisa.
Among the 35 strains of legionellae that had been unidentified at species level, by physiological and serological tests, 27 strains were genetically identified as L. pneumophila, 1 strains as L. feeleii, and another, as L. anisa, but 6 strains were not identified. We found that three strains among the 6 strains belong to a single species of genus Legionella other than the 25 reference species used in this study.

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We examined the sera of 587 hunters in Hokkaido (Japan's northernmost island) for the antibody to Borrelia burgdorferi (B. burgdorferi) by enzyme immunoassay, clarified the conditions related to antibody positivity in these subjects according to region, and studied the effects of factors such as age and lifestyle on the antibody titer. In contrast with an and-B. burgdorferi antibody positive rate of 7.1% in control sera, that in the hunters' sera was 16.0%. Among those positive for the and-B. burgdorferi antibody, the antibody positive rate in sera excluding those testing positive in the serological test for syphilis was 5.5% in the controls, and 15.4% in the hunters, the latter rate being significantly higher (p<0.05). In both hunters and control groups, the antibody-positive rate tended to be higher in older subjects, but the antibody titer showed no correlation with their age, or the duration of their hunting experience. Examination of the hunters' occupations revealed a tendency toward high titers in those engaged in dairy farming. The antibody positivity of those who went gathering edible wild plants was significantly higher than those did not (p<0.05). These observations suggested that the high antibodypositive rate in hunters may have been due largely to the effect of activities other than hunting as sources of infection by Borrelia.

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We determined the minimum inhibitory concentration (MIC) of DR-3355, a newly developed quinolone-derivative antibacterial agent, against clinical isolates of various bacterial species from enteritis patients, and compared them with those of ofloxacin (OFLX), ciprofloxacin (CPFX), nalidixic acid (NA), ampicillin (ABPC), kanamycin (KM). MIC₉₀ of DR-3355 against 94 strains of Shigella spp. and 5 strains of Escherichia coli, 36 strains of Salmonella spp., 22 strains of Vibrio cholerae, 5 strains of Vibrio parahaemolyticus, and 19 strains of Campylobacter jejuni were 0.05, 0.10, 0.0025, 0.39, and 0.78μg/ml, respectively. These values were 1/2 of that of OFLX, and two times of that of CPFX. MIC₉₀ of DR-3355, OFLX and CPFX against C. jejuni were 0.78μg/ml. MIC₉₀ of DR-3355 against isolates from enteritis patients except for Vibrio spp., were 1/30 to 1/60 of those of NA, ABPC, and KM.

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We investigated the influence of the new quinolones for the ability to kill on polymorphonuclear leukocytes (PMNs) in phosphate buffer solution (PBS) and in the urine in order to clarify the in vivo effects of the new quinolones.
Four new quinolones, i.e., norfloxacin, enoxacin, ofloxacin and ciprofloxacin were employed. PMNs derived from peripheral blood in PBS and in hypotonic urine were stimulated with phorbol myristate acetate or opsonized zymosan, and superoxide anion generation of PMNs was measured by means of the chemiluminescence method. The superoxide anion generation of PMNs was significantly enhanced in the presence of 10μg/ml of all new quinolones and significantly suppressed in the presence of 100μ/ml of all new quinolones in PBS and in hypotonic urine.
From these results, it was suggested that no influence of new quinolones on the ability to kill the PMNs might be seen in human blood, however, a great influence might be expected in the urine in the human urinary tract.

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To examine the prevalence of hepatitis C virus (HCV) in haemodialysis patients without blood transfusion in Hiroshima Prefecture, antibody to HCV (anti-HCV) was studied by the Ortho ELISA Kit in sera from 393 consecutive haemodialysis patients and in sera from 510 age and sex matched healthy members of the general population (control). An additional confirmatory test was done by a recombinant immunoblot assay.
1) Anti-HCV was detected in 70 of the 393 dialysis patients and 3 of the 510 healthy controls (17.8% vs 0.6%, p<0.01). Prevalence of anti-HCV in haemodialysis patients sera was increased by the volume of blood transfusion, and even in dialysis patients who had no blood transfusion, the frequency of anti-HCV positivity (9.2%) was greater than the healthy controls (p<0.01). Thus, the major route of HCV transmission in haemodialysis patients without blood transfusion may be via the haemodialysis treatment.
2) The prevalence of anti-HCV increased significantly with the ALT level and abnormal ALT activity of the anti-HCV positive group were significantly greater than that of the negative group. Thus, it is suggested that HCV infection may be an etiologic factor of liver dysfunction in haemodialysis patients.

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Chlamydia trachomatis (C. trachomatis), C. psittaci, and C. pneumoniae are now well established as pathogens of respiratory infections including pneumonia. Serum samples from 223 infants and children with pneumonia, 31 patients with adult inclusion conjunctivitis, 16 parents of babies with neonatal inclusion conjunctivitis and others were tested for IgM antibodies to Chlamydiae.
Diagnostic kits for chlamydial IgM antibodies (SeroELISA^® and IPAzyme^®) have been also evaluated for their diagnostic value. It was found that detection of specific IgM antibodies with SeroELISA has a diagnostic value in chlamydial pneumonias.

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The efficacy of recombinant human granulocyte colony-stimulating factor (G-CSF) in the treatment of Pseudomonas aeruginosa bacteremia in cancer patients receiving intensive chemotherapy was studied retrospectively. In 14 of the 24 episodes of P. aeruginosa bacteremia, which occurred in 23 severely neutropenic patients with hematologic malignancies during a three-year period, G-CSF was given subcutaneously or intravenously at daily doses of 75 & mu;g/body to 200 & mu;g/m² of body surface. Overall, survival at one week after onset was observed in 13 patients (54%). Treatment with G-CSF, however, had no statistically significant association with one-week survival, although a favorable outcome was well correlated with an increase in the neutrophil count during therapy. On the other hand, septic shock and appropriate antibiotic therapy were the major prognostic factors. The frequency of shock was reduced by appropriate therapy, but not by G-CSF treatment. These preliminary findings thus suggested that G-CSF should not be effective in the treatment of neutropenic cancer patients with P. aeruginosa bacteremia. No adverse effects of G-CSF were observed.

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The detectability of mycobacterium in culture by non-isotopic, chemiluminescent DNA probes for Mycobacterium tuberculosis (M. tuberculosis) and M. avium-intracellulare complex (MAC) was evaluated and compared with that by ¹²⁵I-labeled DNA probe for the same mycobacteria. The sensitivity and specificity of the AE-DNA probes for MAC were 97.2% and 100%, respectively, for the conventional method and were both 100% for the ¹²⁵I-labeled DNA probes. The detection limits of the AE-DNA probes tests for both M. tuberculosis and MAC were 10⁵-10⁶ CFU/tube which were almost the same as those of the ¹²⁵I-labeled DNA probes tests.
Because the procedure is simple, rapid (it can be completed within 90 min), and safe (it does not use radioisotopes), it can be easily performed in any clinical laboratory.

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Four cases of respiratory tract infections caused by Corynebacterium pseudodiphtheriticum were reported. The first two patients developed pneumonia with Corynebacterium pseudodiphtheriticum during steroid therapy used against their underlying diseases. The other two patients had acute exacerbation of chronic pulmonary diseases caused by C. pseudodiphtheriticum. These four patients improved by antibiotic therapy. Though nondiphtheria corynebacteria are regarded as “normal flora” when they are isolated from sputum, they should be recognized as potential pathogens.

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A 74-year-old housewife was admitted to the hospital with complaints of high fever and general fatigue. The physical examinations on admission showed no particular findings except for mild hepatomegaly, but labolatory findings showed severe liver dysfunction, active inflammation and negative tuberculine test. On the 4th day, she suddenly complained of severe respiratory distress. A chest X-ray film demonstrated surprising changes in comparison with that taken on admission. On suspicion of adult respiratory distress syndrome (ARDS) associated with miliary tuberculosis (Miliary TB), administration of Methylpredonisolone (1000 mg a day for 3 days) in addition to antituberculous drugs was immediately started. With this therapy she was recovered from such ill condition, but the general exhaustion and slight fever continued. We suspected that her condition might be due to adrenocortical involvement of Miliary TB and hormonal examinations were performed. Unexpectedly, Cushing's syndrome was suspected on the basis of the following; high level of plasma cortisol without normal daily variation, normal ACTH level, an absent response to the Dexamethasone suppression test. Computed tomography revealed left side adrenal mass. During these examinations, renal dysfunction probably due to Miliary TB grew gradually worse and she died of renal failure on the 56th day. Necropsy revealed disseminated tuberculosis involving the lungs and the liver, but the adrenal glands were not examined.

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A 42-year-old male was admitted with subarachnoidal hemorrhage. Dexamethasone 224 mg was used to reduce brain edema. His operation was successful without blood transfusion. No remarkable signs and symptoms were found except HBsAg positive and mild GPT elevation during his admission.
He was discharged on the 33rd day. But 2 weeks later, he felt general fatigue and became worse day by day. He was re-admitted on the 75th day. Several therapies were given but he died of hepatic failure on the 85th day. The autopsy showed liver cirrhosis with massive necrosis.
We believed that the steroid-withdrawal-phenomenon caused exessive immunological response and this process caused his hepatic failure leading to death.

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A 10-year-old male was admitted to our hospital because of lymphadenopathy and fever. Biopsy of a cervical lymphnode demonstrated nodular necrotic foci containing many macrophages phagocyting the karyorrhexic nuclei. He was diagnosed as subacute necrotizing lymphadenitis (SNL) and all the symptoms disappeared spontaneously within a month.
Five year later, lymphadenopathy recurred, and re-biopsy showed the same histological findings and he also recovered spontaneously.
Although the prognosis of SNL is generally excellent and the majority of the patients recover without any treatment, recurrences are very rare, especially in young male patient.

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