Enteropathogenicity and enteroreactive-toxins were examined in 66 strains of Vibrio mimicus and the following results were obtained. Frequencies of enteropathogenic strains judged by the result of suckling mouse tests were 11/13 (85%) for clinical isolates and 37/53 (70%) for fish or environmental isolates. Frequencies of preservation of cholera toxin gene and NAG-ST gene were 2 and 15%, respectively, for 48 enteropathogenic strains, and 0 and 6%, respectively, for 18 non-enteropathogenic strains. Frequencies of production of NAG-rTDH, FAF and hemolysin were 4, 63 and 100%, respectively, for 48 enteropathogenic strains, and 6, 50, and 100%, respectively, for 18 nonenteropathogenic strains. No correlation between serovar and enteropathogenicity was observed in the suckling mouse test. Six out of 12 enteropathogenic strains produced hemolysin in ligated rabbit ileal loop, while 1 out of 12 non-enteropathogenic strains did so under the same condition. A significant inhibition of fluid accumulation in the ligated rabbit ileal loop test with viable cells was noted in rabbits immunized with hemolysin of non-01 V. cholerae. These results suggest that approximately two-thirds of environmental isolates are enteropathogenic and that hemolysin is the most important toxin in the enteropathogenic mechanism of V. mimicus strains.

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Fourty-two febrile episodes of 32 patients with hematologic disease during neutropenia were treated with two randomly assigned antibiotic combinations of either piperacillin plus gentamicin or piperacillin plus aztreonam. Eleven of the 22 febrile episodes treated with piperacillin plus gentamicin and 12 of the 20 febrile episodes treated with piperacillin plus aztreonam responded. Addition of cefamandole to non-responders improved the outcome in 2 of the 16 febrile episodes. Mean nadir leucocyte count, age, sex, and underlying disease were not significantly different in both groups. Side effects were tolerable in both groups, although 1 patient treated with piperacillin plus gentamicin showed severe renal impairment. Piperacillin plus aztreonam is as effective as piperacillin plus gentamicin as an empiric antiotic combination in the treatment of febril episodes with hematologic disease during neutropenia.

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A total of 182 isolates of motile Aeromonas from patients with diarrhea and environmental sources was investigated for hemolytic activity to rabbit erythrocyte and cytotoxicity to HeLa 229 cell. Furthermore, the relation between 0 serogroup and virulence factors, which were lethal to mouse and autoagglutination, were investigated.
There were many strains possessing both the hemolytic and cytotoxic activities in A. hydrophila from overseas traveller's diarrhea, suggesting that these activities were associated with intestinal pathogenecity. Although there was a clear correlation between hemolytic and cytotoxic activities in A. hydrophila from overseas traveller's diarrhea, the correlation was not found in A. hydrophila from domestic cases of diarrhea and A. sobria from overseas traveller's diarrhea. Especially, some A. hydrophila isolates from domestic case of diarrhea produced only hemolysin. These results indicated that there was a difference in specificity between the toxins accounted for hemolytic and cytotoxicactivities, and more than two different toxins were developed.
0 serogroups 11, 34, 14, 16, and 35 in that order were the most frequent serogroups. About half of011 and 034 strains possessed lethal activity to mouse. Autoagglutination phenomenon did not seem to be associated with the lethal activity. In 011 strains, high cytotoxic titer was more frequently found in lethal activity positive-strains than in the activity negative-strains, suggesting that cytotoxicity contrlbuted preferentially to lethal activity to mouse. But such a correlation was not found in 034 strains, so other virulence factors than hemolysin and cytotoxin may be associated with the lethal activity.

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In an in vitro complicated cystitis model, the concentrations of the urinary antimicrobial agents were determined using a computer-controlled automatic urine concentration simulator. The effects on the bacterial count curves showing the presence or absence of PAE in antimicrobial agents were studied by comparing the times required for regrowth to the concentration at the initial inoculation, i.e., effective regrowth time (ERT). The following results were obtained.
1. When beta lactam antimicrobial agents (such as AMPC and CFIX) with no PAE against the gram-negative rods were tested, the ERT of the gram-negative rods were about two hours shorter than that of the gram-positive coccus.
2. When new quinolone antimicrobial agents (such as OFLX) and aminoglycosides (such as ISP) that possess PAE against both the gram-positive and negative organisms were used, there was no difference between ERT of the gram-negative rods and gram-positive coccus.
Therefore, it was demonstrated that the presence or absence of PAE is also reflected in the cell number curve in the case of this in vitro model, more closely related to clinical cases, when the antibiotics is simulated in urinary concentration shifting.

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P. mirabilis and P. vulgaris are the two wellknown species in the genus Proteus. P. myxofaciens and P. penneri are recent additions to the genus. We isolated P. penneri from the pus of a patient with suppurative otitis media and an epidural abscess. The characteristics of the organism, including morphology, staining, physiology and biochemistry, were studied. Clinical microbiological laboratories should suspect P. penneri in the case of as Proteus strain that is negative for indole, salicin and esculin, but otherwise resembles P. vulgaris.
Proteus penneri, formerly known as Proteus vulgaris indole-negative or as Proteus vulgaris biogroup 1, was named by Hickman et al in 1932. Little information about human infection by this organism is available. In 1982, Hickman and co-workers studied 20 strain of P. penneri which were isolated from clinical specinmens (urine, stool, etc.) in the USA. However, its clinical significance, until recently, was unknown. We isolated a strain of P. penneri from the pus of a patient with suppurative otitis media and an epidural abscess on June 10 and 15, 1989. This paper concerns the problems encountered in identifying this organism and its clinical significance.

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Identification of six Legionella species, which we previously identified by serological test as Legionella bozemanii (L. bozemanii), was performed by DNA-DNA hybridization using a commercial DNA-DNA hybridization kit (Kobayashi Pharm. Co., Japan) introduced by Ezaki et al. All strains were identified as Legionella anisa (L. anisa), this being the first identification of L. anisa in Japan. Conventional laboratory tests were performed following the DNA-DNA hybridization. In this study the results of biochemical examination obtained, corresponded closely with those described in previous reports, but the oxidase reaction was very weak and varied according to the age of the culture, indicating the unreliability of this test in our case. All strains examined under long wave ultraviolet (UV) light (366nm) revealed a blue-white fluorescence, the intensity of which ranged from strong to weak. Serological identifications were performed by both the slide agglutination test (SAT) and indirect immunofluorescent assay (IFA). SAT using commercially available antiserum (Denka Seiken., Japan) supposedly specific for L. bozemanii showed cross-reaction between L. bozemanii and L. anisa. Hyperimmune rabbit antisera prepared in this study for both L. bozemanii and L. anisa, from which cross-reactive antibodies were removed by the absorption of each antigen, reacted only with homologous antigens. IFA using a commercially available antiserum and hyperimmune rabbit antiserum previously described, gave positive reactions with each strain.

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Serological diagnosis was examined for deep mycotic infections, especially candidiasis. Candida antigen and antibody researches were made by means of CAND-TEC (RAMCO Co., Ltd) and passive hemagglutination test (PHA) (Roche Co., Ltd), respectively, and simultaneous determinations were made of D-arabinitol, a fungal metabolite and (1-3)-β-D-glucan, fungal parietal component.
1) For normal subjects, 1 of 173 cases (0.6%), 5 of 200 cases (2.5%) and 7 of 157 cases (4.5%) showed <1: 4, 320 × and <11, μmol/1 for CAND-TEC, Candida antibody and D-arabinitol, respectively, and values above these were judged positive.
2) Of the 171 patients with pyrexia refractory to general antibiotic agents, who were treated at our Department and related institutions from November 1988 to March 1990, 41 with obvious Candida infections were CAND-TEC, Candida antibody, D-arabinitol and (1-3)-β-D-glucan positive in 33 (80.5%), 21 (51.2%), 29 (70.7%) and 13 cases (56.6%), respectively. At least one item-positive patient was as high as 97.4%. On the other hand, in the group having no Candida infection at all, 1 of 57 cases on CAND-TEC (1.7%) and 10 of 55 cases on Candida antibody (18.2%) were positive, indicating significant differences from the surely Candida-infectious group.
3) Comparative examination of CAND-TEC and other testing methods revealed a correlation of CAND-TEC with D-arabinitol in cases showing 1: 4 and 1: 8>, but not other significant difference.
4) Also in the respiratory Candida infectious group strongly suspected of Candida infection and the group suspected of urinary Candida infection, CAND-TEC, D-arabinitol and (1-3)-β-D-glucan showed significantly high positivity compared with the group having no Candida infection.
The above results suggest that≥ 1: 4with CAND-TEC is significant for serological diagnosis of candidiasis and that a combined use of CAND-TEC with some other testing method makes early serological diagnosis of Candida infection possible.

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Human papillomavirus (HPV) has been detected on the genitalia without any macroscopicabnormality and the possibility of latent infection of HPV has been suggested. Using Vira TypeTM (Toure Co.), we have detected 7 genotypes of HPV DNA under a high stringent condition on the genitalia of patients with sexually transmitted diseases (STD), who were suspected of having had many sexual partners.
In male cases of STDs other than condyloma acuminatum, the HPV-positive rate of the glans and sulcus coronarius was 4.7%(5/106). In healthy men, the HPV-positive rate was 6.1%(2/33), while in chronic prostatitis cases it was 3.4%(7/205) and in benign prostatic hypertrophy cases HPV was not detected.
In female cases of STDs other than condyloma acuminatum, the HPV-positive rate of uterinecervix was 5.1%(3/58). In pregnant women, the HPV-positive rate was 4.6%(9/197).
With regard to the HPV-positive rate within different age groups of STD and non STD cases, the rate tended to be higher in young people.
After several weeks, follow-up studies were conducted on HPV-positive cases. HPV DNA was detected in one case of 10 male STD cases and two of 10 pregnant women, and the HPV DNA was the same type as at the first examination. However, after 3-4 months, all three of these cases had become negative for HPV DNA.

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A method using polymerase chain reaction (PCR) was compared to an enzyme immunoassay (Chlamydiazyme) for detection of Chlamydia trachomatis by testing a reference strain and clinical specimens. Two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L₂ were used as primers for the PCR. A DNA fragment of 242 by specific for C. trachomatis was amplified by the PCR, when DNA of ≥10² C. trachomatis was used as template for the PCR. A chlamydial antigen was detected by Chlamydiazyme, when ≥2.6×10³ C. trachomatis were applied for the enzyme immunoassay. The PCR method was 26 times more sensitive than Chlamydiazyme in detection of C. trachomatis. The PCR method and Chlamydiazyme were carried out to examine 74 urethral swabs obtained from male patients with urethritis for detection of C. trachomatis. In 45 of 74 specimens, the DNA fragment of C. trachomatis was amplified by the PCR, and in 41 of 74, the chlamydial antigen was detected by Chlamydiazyme. The detection rate of the PCR method (60.8%) was higher than that of Chlamydiazyme (55.4%). The positive coincidence rate of the PCR method to Chlamydiazyme was 100%(41/41) and negative coincidence rate was 87.9%(29/33). The overall coincidence rate between the two methods was high (94.6%). Thus, the PCR method was more sensitive than Chlamydiazyme for detection of C. trachomatis and specific for diagnosis of chlamydial urethritis.

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Treatment of infections by the use of antimicrobial agents should be made essentially in a dose close to the minimally required dose. Acute uncomplicated cystitis in female fits as the subject for a single-dose therapy since it is an infection reactive relatively easily to antimicrobial agents.
Accordingly, an assessment has been made regarding the therapeutic results of the single-dose therapy in 76 female cases of acute uncomplicated cystitis by the use of LVFX 200 mg which is a new quinolone.
The urinary concentration more than MIC₉₀ to Escherichia coli is sustained for about 3 days by this single-dose therapy.
As a result of judging the therapeutic results from the reactions of the three clinical findings of pain on micturition, pyuria and bacteriuria, excellent therapeutic results were obtained with effective rates being 100%(76/76) on the day 3, 93.9%(46/49) on the day 7 and 94.4%(34/36) on the day 14.
The rate of cystitic symptoms which recurred posed no problem, being 12.5%(5/40) up to three months, as investigated by a questionnaire.
As a result of performing close urological examinations such as cystoscopy on six cases with insufficient results or recurrence, we could detect mild underlying conditions which are considered to be intractable factors in the bladder in three cases.
From the above results, the single-dose therapy of acute uncomplicated cystitis in the female by LVFX which is a new quinolone was considered to be an excellent therapeutic drugfrom its characteristics such as its therapeutic results being the same as the conventional therapy by daily administration, excellent drug compliance, low cost, hard selectiveness of resistant strains, less side effects and furthermore it gives the opportunity of detecting a latent and mild underlying condition.

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The author devised a method which permits simultaneous isolation and identification of Chlamydia using a slide chamber with 8 wells. Contaminating bacteria were eliminated by filtration with the menbrane filter. The procedure allowed isolation of Chlamydia pneumoniae and Chlamydia trachomatis from 11 (10.1%) out of 109 and 10 (9.2%) out of 109 otolaryngologic clinical specimens, respectively. The use of HL cells together with Hela-229 cells had been considered essential to isolate Chlamydia from specimens in this field. The conditions for isolating Chlamydia pneumoniae were similar to those for Chlamydia trachomatis. The slide chamber procedure with menbrane filtertreated specimens was suitable for simultaneous isolation and culture of various species of Chlamydia and allowed easy differentiation of Chlamydia pneumoniae from Chlamydia trachomatis with identification by the indirect fluorescent-antibody technique.
Routine examination of clinical specimens using this procedure should be required in the future in order to better understand chlamydial infections.

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Recently, “low dose and long term” erythromycin (EM) treatment has been reported as effective on chronic lower respiratory tract disease, including diffuse panbronchiolitis (DPB). However the effective mechanism of EM is still obscure. In this study, we investigated the effect of EM on intrapulmonary influx of neutrophils by intratracheal injection of lipopolysaccharide (LPS), and thef ollowing results were obtained.
1) The intrapulmonary influx of neutrophils was significantly suppressed (p<0.001) in mice intraperitoneally injected with EM at 5 mg per animal 2 hr before intratracheal injection of LPS (control group: 6.5 ± 0.8×10⁵ vs EM-treated group: 1.7 ± 0.3×10⁵), but not 10 hr before lung challenge. This inhibition was observed at 6 hr after lung challenge, and became maximum with 84% suppression at 24hr.
2) The intrapulmonary influx of neutrophils was not affected when EM was injected intraperitoneally daily for 3, 7, or 14 days, and lung challenge was performed 24 hr after the final administration of EM.
3) The number of neutrophils in the peripheral blood was not affected by EM.
These results suggest that EM treatment impairs the capacity for pulmonary inflammation by reducing, at least in part, the migration of neutrophils to inflammatory sites.

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In Chiba Prefecture, the first patient of infection with spotted fever group Rickettsia was found in 1987, thereafter nine patients were detected serologically by the end of 1990. Patients were found in the villages of Amatsukominato, Katsuura and Ootaki, which were located in southern part of Chiba Prefecture. The illness occurred from June to October. On the other hand, patients with Rickettsia tsutsugamushi were seen from October to September. Difference of prevalent seasons of these two types of rickettsiosis is important to make a clinical diagnosis and serological identification of this spotted fever group rickettsiosis. Antibody of the patients showed the highest titer to YH strain of Rickettsia and showed high cross-reactivity to other spotted fever group rickettsiae. For the diagnosis the patient serologically, it was confirmed that any strain of spotted fever group Rickettsia were useful.

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The clinical features of tracheobronchial tuberculosis are distinct from those of pulmonary tuberculosis in some aspects. Streptomycin (SM) is claimed by some investigators that it has a tendency to promote the development of bronchial stenosis due to scarred healing of the involved endobronchial mucosa, and, for that reason, they recommend not to use it in treatment of bronchial tuberculosis. In some patients with pulmonary tuberculosis, who have renal orhearing disturbance, SM avoided. Yet unless SM is used improvement of tuberculosis could be delayed.
It is the purpose of the present investigation to point out that aersol therapy of SM is useful for patients with respiratory tuberculosis. Furthermore, we wish to demonstrate that we can safely treat the patients with respiratory tuberculosis who suffer from renal function or hearing disorder by use of aerosol therapy of SM.
Prior to clinical application of the inhalation therapy, we comfirmed that the therapy was not harmful. Serum concentration of SM, when inhaled, was measured in 9 volunteers. Before and after administration of SM aerosol, spirograms were examined in 4 volunteers, nevertheless, no special abnormality was recognized. It seemed that serum concentration of SM after the administration was too low to evoke adverse reactions (less than 3.0 γ). In 6 patients with pulmonary tuberculosis, blood gases were measured and no obvious change was observed. As a result, it was demonstrated that the endobronchial tuberculous lesions improved faster when treated by inhalation of aerosolized SM as compared with the conventional injection therapy, without evoking any apparent adverse reactions. In some of the patients with pulmonary tuberculosis, it seemed that the therapy was useful.
We conclude that the inhalation therapy with aerosolized SM is very useful for the patients with tracheobronchial tuberculosis and the therapy is valuable in treatment of thepatients with pulmonary tuberculosis.

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Sterile pleural effusion was induced by intrapleural administration of turpentine, and empyema was induced by direct intrapleural inoculation of bacteria in rabbits. Experimental models of anaerobic thoracic empyema were successfully made in this study, using Escherichia coli, Peptostreptococcus asaccharolyticus, Bacteroides fragilis, using the following methods;(1) a single bacterium inoculation of, a) E. coli, b) P. asaccharolyticus, c) B. fragilis, (2) simultaneous inoculations with an aerobe and an anaerobe, a) E. coli+P. asaccharolyticus, b) E. coli+B. fragilis. (3) an anaerobic inoculation 5 days after an aerobic inoculation, a) P. asaccharolyticus inoculation 5 days after E. coli inoculation, b) B. fragilis inoculation 5 days after E. coli inoculation. The developments of several parameters (sialic acid levels, glucose levels, oxidation-reduction potential levels, bacterial counts and WBC counts in the pleural fluid and so on) were measured in each group.
There was little difference of the rate of incidence of empyema between each group. Empyema was highly induced even by a single anaerobe inoculation. The following tendencies were observed in development of parameters:(1) sialic acid levels and oxidation-reduction potential levels were decreased only in the conditions being associated by B. fragilis infections. (2) in mixed infections of E. coli and anaerobes, bacterial counts of E. coli increased in number in the late phases of the infections probably by the influences of anaerobes. The experiments of groups (2) and (3) were designed for demonstrations of biphasic infections between aerobes and anaerobes, but it turned out that quite unexpected results were seen in these models.

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Pasteurella multocida is a gram-negative short rod-shaped bacteria, which is a part of the indigenous flora of the oral cavity of many animals other than man. The number ofreports on cases of infections with this bacterium due to animal bites and/or scratches, bacterial infections of the respiratory tract, sepsis due to this organism and death caused by the bacteria have been increasing in recent years.
We investigated P. multocida in the hair and oral cavity of 3 dogs and 29 cats according to the classification of Mutters et al. We also studied the relationship between the carrier rate for Pasteurella in the oral cavity and kissing of pets in 24 pet owners (3 dogs and 11 cats).
No P. multocida was isolated from the hair of neither dogs nor cats. One strain of P. multocida subsp. multocida and two strains of P. stomatis, were isolated from the oral cavity of dogs, and 35 strains of Pasteurella were isolated from the oral cavity of cats. Two strains ofP. multocida subsp. multocida, whose biochemical properties were different, were detected in the oralcavity of one cat. In three cats, Pasteurella other than P. multocida subsp. multocida was isolated from the same oral cavity. No Pasteurella was detected in the oral cavity of 19 pet owners who had not kissed their cats, whereas P. stomatis was isolated from the oral cavity of one of 2 pet owners who had kissed their cats and in 2 of 3 pet owners who had kissed their dogs (the same bacteria was isolated from a dogthat was being kept by some of these positive pet owners). When one of these three pet owners refrained from kissing the pet animal, and was re-examined one month later, no bacteria were detected. Therefore the infection was considered transient.
The above results suggest that kissing pets can cause infections in humans.

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In 17 patients of genital herpes virus infections, herpes simplex virus (HSV)-specific IgA antibodyresponses were analyzed using an enzyme-linked immunosorbent assay (ELISA) and the absorption ofsera with Streptococcus pyogenes, AW43 and AR1, which bind monomeric IgA (m-IgA) and IgGantibodies, respectively. The ratio of polymeric IgA (p-IgA) to total IgA was calculated from theformula, the p-IgA index=IgA activity after absorption/IgA activity before absorption. In earlyconvalescent-phase sera from primary cases, the p-IgA indices were 0.5-0.9, whereas in recurrent orprovoked cases, they were lower than 0.2. There results indicate that the p-IgA index may be useful forrapid and simple differentiation of primary from non-primary HSV infections.

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A 53-year-old male visited our hospital due to nasal obstruction persisting for 6 months and constant rhinorrhea. Pasteurella multocida subsp. multocida was isolated from his nasal discharge and lavage fluid of the maxillary sinus, and also from the oral cavity of the dog he kept. The bacterial strains isolated from the patient and dog were identical in terms of biochemical properties, and drug sensitivity. Although serotype was different, the strain from the patient showed (A: 6) and that from his dog showed (A: 5). The microorganism is not present in the general environment. The patient had contact with his dog such as he kissed it frequently, gave it food with his chopsticks et al. From the mouth of the people who kiss one's dog, we detected Pasteurella of the same character of bacteria as from the mouth of the dog. We detected two Pasteurella multocida of different character from only one mouth of a cat. Pasteurella multocida was checked in only one colony for sero type. Sero type A is the popular type for dogs and cats. The above suggest that their was a high possibility that the Pasteurella multocida subsp. multocida found in the patient was from his dog.
In Japan, the incidence of Pasteurella multocida subsp. multocida infection has been increasing. In 1969, the Japanese Ministry of Health and Welfare officially communicated this infection as a zoonosis to related institutions. At both medical and surgical departments, wither the patient keeps a pet should be confirmed during interview, and guidance of pet keeping methods is important in some cases from the aspect of clinical bacteriology.
We await the establishment of a general institution which collects and analyzes data on various zoonoses in various fields, and formulates preventive measures.

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