Antigenic relationships among the 11 strains of Japanese encephalitis (JE) virus were analyzed by using monoclonal antibodies (NARMA) against Nakayama-RFVL strain in the hemagglutination inhibition (HI) and neutralization (Nt) tests. Of the 14 JE virus specific HI antibodies, all except NARMA 5 showed the Nt activities against the homologous strain. The HI and Nt titers of these antibodies were not parallel. They consisted of the following characteristics antibodies: NARMA 3, a species-specific antibody with HI and Nt activities against JE virus; NARMA 13, a species-specific antibody with HI antibody; NARMA 6, a Nakayama strain-specific antibody with HI and Nt activities; NARMA 5, a Nakayama strain-specific HI antibody; 10 other antibodies, all of the intermediate type.
The eleven strains of JE virus were classified into the following antigenic groups:
I-1, Nakayama-RFVL and Nakayama-Yoken strains.
I-2; Nakayama-Yakken strain.
II; G-1 late, JaGAr 01 and JaGAr 02 strains.
III-1, Kamiyama, Mie 44-1 and Kumamoto 80679 strains.
III-2; 691004 strain.
IV; Muar strain.
These results were in reasonably good agreement with the antigenic classification of the JE virus strains performed by the HI test using 5 monoclonal antibodies (NARMA 1, 4, 5, 8 and 13), although slight antigenic differences were found among some strains of the same group.

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Safe and simple methods for sampling sputum, which can be applied to general clinical fields, are desired for the survey of causative bacteria in pulmonary infection. In 171 patients with pulmonary disease, local bronchial sputum was sampled with a brush used for the diagnosis of pulmonary carcinoma. The cultures of the samples thus obtained (bronchial secretions and exudates, abbreviated to BS) and of simultaneous, expectorated sputum were compared, and the following results, with regard to the degree of contamination associated with this method, the conditions for determining the causative bacteria of the isolated potential pathogens, etc., were obtained.
1) Agreement of the component species of bacterial flora between BS and simultaneous expectorated sputum was found in 26.9% of the patients.
2) The colony counts of Neisseria isolated from BS were judged as ++ in 5.3% and as slight or less in 65.5%. With expectorated sputum, corresponding counts were assigned to ++ or +++ in 65.5%. A similar distribution of colony counts was obtained for α-Streptococci. Thus, it is presumed that this sampling method is associated with minimal contamination of BS by indigenous bacterial flora in the upper airway.
3) The colony counts and the prophlogisticity of the pathogens were compared between pulmonary parenchymal infection and airway infection. When the colony counts of bacteria isolated from expectorated sputum were rated as ++ or +++, 31.3% of the bacteria were causative in pulmonary parenchymal infection, and 100% in airway infection. When the colony counts of bacteria isolated from BS were ++ or +++, 100% were causative in both infections.
4) The frequency of causative bacteria isolated from purulent sputum was markedly higher than in other specimens.
On the basis of the above results, the position of this method in the survey of causative bacteria in pulmonary infection is also discussed herein.

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We examined the prevalence of chlamydial infection in a population of pregnant women and observed their infants to determine the risk of development of infection.
In this study cycloheximide-treated McCoy cell was used for isolation of C. trachomatis.
C. trachomatis was isolated from cervical swab specimens of 3 (3.6%) of 82 pregnant women in the S.C. hospital, 1 (0.9%) of 108 pregnant women in the D.A. hospital and 1 (1.6%) of 12 pregnant women having antibody to C. trachomatis in the S.T. hospital.
Development of chlamydial infection was proved 2 of 5 infants born to the infected women.

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As we have treated children with malignancies for combination chemotherapy of multiple anticancer agents, severe neutropenia has been often recognized. Once they become severe neutropenia, they often have high fever, but we cannot find out the etiology of their fever. Most researchers recognize that the etiology of this fever is septicemia, but they cannot almost detect pathogenic microorganisms by conventional blood culture. In this paper, I discussed that the possibilities of improvement in blood culture method, and I obtained next results.
1) I evaluated whether in vitro antibiotics, which were present in blood, have influence on the detection of microorganisms by blood culture. In artificial bacteremia 3.0×10^°FU/ml of S. aureus was inhibited by 1.0μg/ml of gentamicin. Therefore, if bacteremia level is less than 3.0×10^°FU/ml of S. aureus, blood culture, which was done within 6 hours after drip infusion of 2 mg/kg of gentamicin, might not be positive. As in bacteremia of E. coli the growth of bacteria was inhibited by more than 10.0μg/ml of gentamicin, the results of blood culture were not influenced by conventional dosage of gentamicin. I evaluated whether in vivo antibiotics which were present in patients' blood have influence on the detection of pathogens by blood culture. Blood culture which was done after 2 hours after administration of antibiotics did not have influence on the growth of microorganisms.
2) I evaluated that blood volume from 5% to 30% inoculated into media has influence on the detection of microorganisms. At both blood of leukemic patients with leukopenia and normal healthy adults, the detection rates of bacteria made no difference. Namely it took more than 24 hours to recognize the growth of bacteria at 5% inoculated blood volume, and even 30% inoculated blood volume did not inhibit the growth of bacteria.
3) I tried to separate bacterial pathogens from all antibacterial properties of the blood, including antibiotics, complement, phagocytes and antibodies by centrifugation. But controlled culture with no treatment had a trend to growth more rapidly, I could not obtain more sensitive culture method by centrifugation.
4) I tried to make the layer in which almost all of microorganisms were present by centrifugation. At the speed of 3, 000 rpm, bacterial pathogens remained in the layer of blood cells. All microorganisms could not be sedimented at the bottom of blood cells layer.

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On the course of treatment for children with malignancies, we have often experienced so-called sepsis like symptomes. From these cases, it is very difficult to detect pathogenic microorganisms by conventional blood culture methods. In this report, I discussed about the Lysis-Filtration blood culture technique which was first described by Zierdt et al. and I tried to reform some important points.
The following results were obtained.
1) The optimal condition of Lysing Solution (L.S.) was determined for both bacterial toxicity and lysing capability. The L.S. was satisfied by the condition with concentrations ranging from 0.05 to 1.0% of Protease, ranging from 0.1 to 2.0% of Tween 20 and ranging from 6.0 to 10.0 of pH.
2) There were no specific changes in the nature of L.S. when stocked at 4°C for one year.
3) As the antibactericidal factors of the blood are removed before cultibation, the detected CFU of bacteria was correlated with inoculated blood volume. This is the reason why L-F method was more effective as the lower level bacteremia with less than 1 CFU/ml.
4) The L-F method is higher than the broth method regarding the detection rate of bacteria in artificial bacteremia by S. aureus (p<0.05). When the bacterial level was less than 1 CFU/ml, the significance was getting higher (p<0.01). At the artificial bacteremia by E. coli, the same tendency was recognized.
5) Both culture methods were used at the same time for the patients of malignancies who had febrile episodes. 9 cases of 163 (5.5%) L-F method and 8 cases of 162 (4.9%) conventional method detected bacteria, but no significance was recognized between the detective rate of both ways.

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Anaerobes isolated from 804 clinical specimens, considered not to have been contaminated by normal flora, during the five years between January 1978 and December 1982 at the Medical College Hospital of Oita and Nagasaki University Hospital, and aerobes isolated with the anaerobes were analyzed according to the species and frequency, in order to obtain data on the pathogenesis of anaerobic infection and its treatment.Anaerobes alone were isolated from 34.5% of the specimens, and they were isolated together with aerobes from 75.5% of the specimens.Of 940 strains of anaerobes, the frequency of isolation of gram-negative rods including Bacteroides fragilis was highest (47.6%), followed by grampositive cocci (21.5%), non-spore forming gram-positive rods (20.6%), gram-negative cocci (7.1%) and Clostridium species (3.2%), in that order.Of 1, 183 strains of aerobes isolated with anaerobes, the isolation frequency of the family Enterobacteriaceae was highest, 44.2%, followed by gram-positive cocci (41%), glucose non-fermentative gram-negative rods (7.4%), glucose fermentative gram-negative rods (3.8%), gram-negative cocci (3.1%) and grampositive rods (0.5%), in that order.The frequency of Escherichia coli isolation was highest (38.9%), followed by α-streptococci (27%), Streptococcus faecalis (21.3%), coagulase-negative staphylococci (16%), Klebsiella aerogenes (13.3%) and Pseudomonas aeruginosa (13%), in that order.There were characteristic relationships among the infection site, the isolates and isolation frequency of the species.The frequency of isolation of B.fragilis was significantly correlated with that of E.coli and S.faecalis, which were isolated along with B.Fragilis (by x²-test: p<0.01, p<0.05, respectively).However, this tendency was not definitely observed for P.aeruginosa or Klebsiella species.

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The survey on possible pathogenic bacteria and their susceptibilities to antibiotics in collaboration with 11 departments of oral surgery was made, and 326 strains of bacteria were isolated from the closed pus of 156 patients with odontogenic infections.The results obtained were as follows:
1.Mixed infection cases were dominant and the isolation rate of anaerobic bacteria was relatively high.These patterns were not related to the type of disease such as periodontital infection, pericoronitis or osteitis.
2.α-Streptococcus was the most major isolates suggesting important causative bacteria. The other bacteria isolated in higher frequency were β-Streptococcus, B.catarrhalis, Bacteroides, Peptococcus and anaerobic Streptococcus.
3.Antibiotics used for the susceptibility test were a new macrolide TMS-19-Q (TMS), josamycin (JM), midecamycin (MDM), erythromycin (EM), ampicillin (ABPC) and cephalexin (CEX).All antibiotics except for CEX showed good antibacterial activities to the isolates without high degrees of resistance.Particularly TMS, EM and ABPC exhibited strong activities though moderate degrees of resistant strains to ABPC were observed in Bacteroides.
In conclusion, like β-lactam antibiotics, macrolide antibiotics such as TMS or EM are suggested to be useful for acute odontogenic infections.

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A formalin-inactivated Japanese encephalitis virus (JEV) vaccinewas inoculated into mice treated with cyclophosphamide (CY) at various intervals, and its effectson the antibody response were investigated. CY treatment in a few days after immunization delayed the appearance of the HI and NT antibodies in sera and decreased the maximum levels of the antibody titers.As for the immunoglobulin classes of the HI antibody, however, the pattern that IgM appeared first then IgG became predominant was the same as that observed in non-CY-treated mice.
In another experiment, live JEV was inoculated into CY-treated mice after immunization with formalin-inactivated vaccine.A few mice died of encephalitis after a single injection of CY and more mice after double injections suggesting that the delayed antibody response at the early stage of JEV infection might lead to serious consequences in experimentally infected animals.
After a single injection of CY, peritoneal exudate cells (PEC) decreased in number during day 4 through day 7, and random migration of PEC was suppressed during day 1 through day 8 suggesting that macrophages were also damaged by CY at a dose of 240μg/g of mouse body weight.

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Several dozen cholera cases occur each year in Japan. Most of these are mild cases due to Vibrio cholerae eltor. This report presents one case of El Tor cholera, contracted from imported perishable foods and resulting in acute renal failure due to dehydration from severe diarrhea. This case was complicated by rhabdomyolysis at the onset, and bacteria were present in the gallbladder for up to 2 months after the disappearance of symptoms. This case was remarkable. in its clinical course, which differed from those for cholera usually seen in Japan.

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Although it has been though that hepatitis B virus (HBV) can be transmitted by sharing a razor, there have been no report on such a case. We found a case (case A) may have contracted hepatitis B by sharing a razor of her friend who was positive for HBsAg and HBeAg
In January 1983, a 15-year-old, female, junior high school student in Okinawa, Japanan area in which hepatitis B is endemic-developed fulminant, type B hepatitis. We investigated the route of HBV transmission. None of the four other members of her immediate family was positive for HBsAg but at the school she was attending, 14 of the other 341 students (4.1%) and four of the 20 teachers (20.0%) were positive for HBsAg, Eight of these 14 students, but none of the four teachers, were positive for HBeAg. Quetioning revealed that, on overnight school trip two months before she became ill, case A had shared a razor of one of the eight HBeAg-positive students. Case A used the razor to shave her legs immediately after the carrier had used it. Cases A said that they had no attempt to sterilize the razor after the carrier used it. Case A denied habing any other close contact, such as sexual contact or sharing of a tooth brush, with anyone in the six-months period preceeding the onset of her illness. The carrier's HBsAg subtypes was adw; case A's subtype could not be detected because of the low HBsAg titer in her serum.
From the findings described above and the fact that blood positive for HBeAg is thought to be highly infectious, we concluded that case A had possibly contracted hepatitis B by sharing a razor contaminated with HBV. We believe that HBsAg carriers should be taught how to avoid transmitting the disease to others.

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