We identified the serotypes and genome patterns of 100 strains of Pseudomonas aeruginosa (P. aeruginosa) that had been isolated from patients who were admitted to the hospital of the Fukui Medical School between 1992 and 1995. A monoclonal diagnostic kit was used to identify the serotypes. Genome patterns were determined by pulsed field gel elecrophoresis (PFGE). Serotypes A, B, C, D, E, F, G and I exhibited distinct genome patterns. Differences in genome patterns were also observed in strains of serotypes E and G, depending on the types of clinical samples collected and/ or the area of the hospital from which they were isolated. Many of the multiple antibiotic-resistant strains of P. aeruginosa exhibited serotype E. The genome pattern differed between strains that were susceptible vs. resistant to multiple antibiotics. The latter strains exhibited similar genome patterns regardless of their origin. These findings suggest that analysis of genome patterns is important for identifying the origin of a nosocomial infection caused by P. aeruginosa, serotype E.

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Biophila wadsworthia is a recently recognized nonspore-forming anaerobic gram-negative rod and is reported to be associated with various infections such as gangrenous perforated appendicitis, peritonitis, osteomyelitis and bacteremia. Although the isolation of B. wadsworthia seems to be facilitated by using Bacteroides bile esculin (BBE) agar, the reliable scheme of identification of this species has been not published. This study was conducted to find simple, rapid, and reliable measures for identification of B. wadsworthia. A total of 32 B. wadsworthia-suspected clinical isolates as well as B. wadsworthia WAL 7959 and Desulfomonas pigra DSM 749, which is the species that shows bacteriological characteristics similar to B. wadsworthia, were used. Through the study of various biochemical and enzymatic tests and culture on selective media, it is indicated that a nonspore-forming, gram-negative anaerobic rod that is nonmotile, forms “blackeyed” colonies on BBE agar, and demonstrates strongly positive catalase test, positive acid phosphatase test, and no-growth on Baceterides medium or to be susceptible to colistin (10μg disk) can be identified as B. wadsworthia.

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Genotyping of 67 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) separated from patients in a hospital in Mizusawa City in 1994 and 1995 was studied by pulsed field gel electrophoresis (PFGE) and arbitrarily primed-polymerase chain reaction (AP-PCR). Two main genotypes were observed by PFGE, and more than 70% of the 67 MRSA isolates produced coagulase type II. One group diverged well and gained higher tolerance in 1994, but was not isolated in 1995. The other group was continually isolated during the two-year period and showed moderate tolerance in 1994, and higher tolerance in 1995. AP-PCR was able to classify the genotypes of MRSA into 6 subgenotypes under the present conditions, which supported the results obtained by PFGE. These results suggest that AP-PCR could become a convenient and useful typing method by improving both sequence and length of a primer.

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In order to clarify the possibility of feco-oral transmission of Helicobacter pylori, we have employed an immunomagnetic beads (IMB) separation technique and tried to isolate _{H. pylori} from fecal samples. Microaerophilic cultivation and PCR of H. pylori were positive when H. pylori (10³ cfu) was suspended in Hanks' balanced salt solution (HBSS) for the IMB separation technique. In contrast, when a lower number of H. Pylori (10² cfu) was suspended in HBSS, 2nd PCR was positive but cultivation was negative. After IMB separation technique from fecal sample containing H. pylori (2-10⁴ cfu), 20 cfu of H. pylori was recovered by culture. In this case, 1st PCR was negative but 2nd PCR was positive. From a fecal sample of a patient infected with H. pylori, H. pylori was detected only by combination of IMB separation and PCR, although H. pylori was not recovered by culture. These results indicate that IMB separation technique followed by cultivation or PCR would be useful for estimation of the presence of H. pylori fecal samples.

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We have characterized clinical and diagnostic features in 18 cases of Legionella pneumonia. Age average of patients was 62.0 years old (male: female= 14: 4) and underlying diseases were observed in 12 patients. Legionella pneumonia were diagnosed in 3, 5, 8 and 9 cases by culture, serum antibody measurement, urinary antigen detection and PCR, respectively. Sixteen cases were caused by L. pneumophila, while the other 2 cases were L. bozemanii pneumonia and L. pneumophila or L. dumoffii pneumonia. Chest X-rays of those patients showed multiple pneumonia shadows in 14 cases, alveolar shadows in 10 cases, pleural effusion in 5 cases. Blood-gas analysis on admission indicated hypoxemia in all cases with abnormal A-a D0₂. Laboratory findings showed abnormal data in WBC, CRP, LDH, CPK and liver function tests (ex. GOT, GPT) in most cases. Serum antibody testing showed positive by 5 weeks after onset of pneumonia, but 10 cases of Legionella pneumonia diagnosed by other techniques were judged to be negative. In urinary antigen detection test, 6 and 2 cases showed positive 1 and 4 weeks after onset of pneumonia, respectively. Macrolide antibiotics were administered in all cases during the episode, but delay of macrolide administration was observed in 3 of 4 cases of dead outcome. Serum antibody measurement, urinary antigen detection and PCR, in addition to culture of bacteria, may be required for exact diagnosis of Legionella infection.

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Approaching the problem of how close the sporadic diarrhea diseases correlate with epidemics of infectious diarrheas, including food poisonings, bacteriological studies were made on the characteristics of sporadic diarrheas in the past 10 years and analyzed.
In collaboration with physicians, mostly pediatricians, in Oita district during 1985 to 1986 and 1989 to 1996, a total of 1707 specimens was obtained from diarrhea patients, suspected of bacterial cause. We have isolated 717 strains from 670 specimens (ca 40% of the total). The majority were; Campylobacter, Salmonella and EPEC with the rate of about 40, 24 and 23% respectively. On the other hand, isolation of Vibrio spp. and Staphylococcus were very few.
The yearly trend of detection rates of such major agents were as follows: Campylobacter had the highest rate in the early period, 1989, but thereafter tended to decrease. In contrast, Salmonella continued to increase in rate through the study period until the present when it exceeds that of Campylobacter. This alternation is due primarily to the rapid increase of S. Enteritidis since its recent appearance. VTEC stays at a low rate with a slow increase showing no prevalence so far.
From the above results the following may meet with attention. Vibrios and Staphylococci, known as popular agents for food poisonings, seem unlikely to correlate with sporadic diarrheas. Instead, the evidence that recent food poisoning is often represented by Salmonella, especially S. Enteritidis, suggests that epidemics of infectious diarrhea may be attributed to the background of common sporadic diarrheas.

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We studied the experimental urinary tract infections (UTIs) on compromised mice with Esherichia coli to elucidate the effects of three factors (bacterial strains, doses of hydrocortisone, infectious periods) on the bacterial colonization in the bladders.
We selected three strains from the stock of E. coli isolated from the urine or feces of patients with lower UTI, because each of the three strains showed a tendency in colonization in previous experiments: One strain, which was named H70, showed a relative slight and intracellular colonization; the H99, an extracellular and the K88, both intracellular and extracelluar growths which were large.
The rate of bacterial microscopic recovery from the bladders of mice inoculated strains H70, H99 and K88 were 25.0, 41.7 and 79.2% respectively. And the recovery from the mice inoculated cortisone 0, 2, 4 and 8 mg/mouse were 38.9, 55.6, 55.6 and 44.4% respectively. The detections of various colonizations at 3, 6 and 24 hr after the bacterial inoculation were that extracellular types were 45.8, 20.8 and 20.8%, intracellular types were 4.2, 16.7 and 12.5%, simultaneous colonizations were 4.2, 16.7 nad 4.2% and the detections of at least one of them were 54.2, 54.2 and 37.5% respectively.
Thus, the global findings were that extracellular growth of the bacteria were predominant at first, then the bacteria invaded the mucosal cells and the intracellular bacteria stayed after the voiding of extracellular bacteria. Intracellular colonizations seemed more strongly interacting with hosts than extracelluar types, because cortisone enhanced the intracellular colonizations more strongly than the others.
However, it seemed that urinary tract infections were not simple events but the results of the interactions of the bacterial strains and hosts, because the locations and degrees of the bacterial colonization and the effects of hydrocortisone on them were different in each bacterial strain and infectious period.

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The phagocytic bacterial activity on Streptococcus intermedius and Streptococus oralis, clinical isolates from patients suffering from severe oral surgical infections, was evaluated by using sera and neutrophils of patients who had been treated with polyethylene glycal treated human normal immunoglobulin (Venoglobulin-IH^®).
The phagocytic function of the neutrophils against the two strains, approximately 10 cells/ neutrophil, was unrelated to the administration of Venoglobulin-IH. The bactericidal function of the serum and neutrophils on the third day following Venoglobulin-IH administration, on the other hand, rose to 10 times that of the sera and neutrophils of patients with mild infections. The agglutinin titer of Venoglobulin-IH against S. intermedius and S. oralis were 32 and 16 times, respectively, indicating an evident rise in these titers against the clinical isolates in comparison with the reference strains of the same organisms.
These findings indicated that the oral streptococci that are isolated from patients with grave infections are associated with an antigenicity that is different from the normal bacterial flora; and these organisms are more labile to phagocytosis by neutrophils that were obtained from patients who have been treated with Venoglobulin-IH.

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To obtain the basic data on the route of infection of intestinal parasites as zoonosis, we examined the feces from 87 main-land raccoon dogs, all of which were kept in the Yokohama Kanazawa Zoo (Kanagawa) between 1994-1996, for detection of intestinal parasites and evaluated the detection rates. Intestinal parasites were detected from 72 of the 87 raccoon dogs (82.8%). Among the parasites, Ancylostoma spp., Trichuris vulpis, Ascaridoidea and Isospora spp. were most commonly detected. Of the 72 raccoon dogs with parasites, 43 (59.7%) had mixed infection of more than two species of parasites. Since Ancylostoma spp., Ascaridoidea and Isospora spp., which are important causes of human diseases, were detected at relatively high rates, it was suggested that the raccoon dog could be a source of human infection of intestinal parasites.

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This report concerns a male patient aged 25 years, diagnosed at the age of 12 years as suffering from chronic granulomatous disease. This patient had p47-phox deficiency. He was admitted to this hospital because of fever and dyspnea accompanied by right spontaneous pneumothorax. He failed to respond to medical treatment. He died from respiratory failure four months after admission. Autopsy demonstrated pigmented lipid histiocytes characteristic of CGD. These characteristic pigmented cells were distributed in the spleen, liver, lymph nodes and in the smal intestine. As for the nature of the pigment, lipofuschin-like compound were identified. Granulomatous component was seen in the mucosa of the stomach obtained by operation.
The presence and characteristic distribution of such pigmented macrophages in tissue in young adults may suggest the diagnosis of CGD.

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Schistosomiasis haematobium is a common infectious disease in certain countries in Africa and western Asia. However to our knowlege, only three Japanese patients have been reported to have this disease in Japan in the 1990's, and in all three cases the disease was contracted outside of Japan. Recently, we treated a non-symptomatic Japanese patient infected with Schistosoma haematobium who had recently returned from Africa. The purpose of this report is to emphasize the importance of parasitological investigation for people who have visited or resided in tropical developing countries, even if no symptoms of infection are evident.

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Infective endocarditis caused by Kingella denitrificans occurs rarely. A review of the literature reveals only 6 cases of endocarditis caused by the bacillus. K. denitrificans is normally a commensal of the upper respiratory airways, may exceptionally be responsible for endocarditis. A case of possible prosthetic endocarditis caused by K. denitrificans is presented.
A 78-year-old male with Type II diabetes was admitted to the hospital complaining of fever, a sore throat and arthralgia. He underwent replacement surgery of a St. Jude medical prosthesis for aortic stenosis at the age of 75. The only physical findings at admission were a temperature of 38.2°C and murmus of mild mitral regurgitation. The liver and spleen were not palpable, and there were no skin or eye lesions. Laboratory findings were as follows: white blood cell count 9500/μl with 77% neutrophils, erythrocyte sedimentation rate 71 mm/h (Westergren), blood urea nitrogen 50.2 mg/dl, serum creatinine 1.7 mg/dl and C-reactive protein 22.2 mg/dl. The Gram-negative bacillus isolated from the blood Was identified as K. denitrificans by the identification system, namely ID test·EFN-20 rapid^®(Nissui, Japan). Although an echocardiogram detected no vegetation, infective endocarditis was diagnosed because the same bacillus was detected by separate blood cultures and an obvious source of infection was not found other than the prosthetic valve. Initial treatment was flomoxef, which was changed to Ampicillin 2g/day after K. denitrificans was identified. Ampicillin was continued for 6 weeks. The clinical course was good and he did not require further surgery. He has been afebrile for 2 years after completing treatmen.
This case represents the first report of prosthetic valve endocarditis caused by K. denitrificans in Japan.

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A 75-year-old female with diabetes mellitus, who was born and lived in West north Kyusyu, was admitted to our hospital, because of unconsciousness and loss of appetite. The physical examination showed neck stiffness and a high fever. The laboratory data showed accentuation of inflammatory reaction and azotemia and positive HTLV-1 antibody. The spinal fluid showed increase of cell count and amount of protein. A stool and sputum smear revealed rhabditis form larvae of the nematode. Antibiotics and ivermectin were administered for the bacterial meningitis and hyperinfection of the strongyloides, respectively. Consequently, meningitis and strongyloidiasis improved. It was considered that the patient was infected with strongyloides from her husband who served in the army during World-War II, and hyperinfection of strongyloides resulted from the immunosuppressive state of diabetes mellitus. Ivermectin, an anti-strongyloides agent, was effective, and no side effects were seen. However, the therapeutic resistance in this case was associated with the positive HTLV-1 antibody.

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