Group B Streptococcus were isolated from 1404 pregnant out-patient women in the Department of Obstetrics and Gynecology, St. Marianna University School of Medicine, from June 1, 1992 to May 31, 2001. Serotype of 187 (13.3%) fresh isolates of GBS was determined by using hemolytic streptococcustyping immune sera (Denka Seiken, Tokyo, Japan). With these strains there were 59 (31.6%), 46 (26.6%), 19 (10.2%), 16 (8.6%), 16 (8.6%), 12 (6.4%) and 3 (1.6%) indicating that their types were VIII, VI, III, Ia, V, Ib and II respectively. Also, bacterial agglutination reaction was employed for detection of titer to GBS in pregnant women serum. Positive reaction showed in 31 (57.4%) samples but 23 (42.6%) samples were not seen in this assy. It is assumed that the high antibody levels in pregnant women serum plays an important role as an in vivo defence factor in neonatal infection by GBS.

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Three patients developed acute Q fever after returning from an inspection tour of farms and abattoirs to Australia. Serum levels of antibodies to Coxiella burnetii and the presence of C. burnetii-DNA in blood samples were examined for more than 100 days. Four-fold raises of IgM and IgG antibodies against C. burnetii phase2 were observed within the first three weeks in all the three cases. Maximum titers of IgM and IgG antibodies were 1, 024-2, 048 and 512-4, 096, respectively. According to the temporal diagnostic criteria of acute Q fever in the convalescent serum: the IgM titer of≥64 and IgG titer of≥512 against phase 2, patient A, B and C were determined to be antibody positive for 45, 199 and 122 days, respectively. The result suggests that this standard is practical and reasonable for diagnosis of acute Q fever. C. burnetti-DNA was detected in the sera and buffy coat samples of patient A who developed high fever, severe thrombocytopenia and liver disfunction, but not in those of patient B and C. This study provides useful information for optimization and standardization of Q fever diagnosis in Japan.

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Fifty-six levofloxacin-susceptible strains of streptococcus pneumoniae were isolated from various clinical material in July, 2002 from June, 2001, examined antimicrobial susceptibility testing of levofloxacin and sparfloxacin, and performed analysis of gyrA gene and parC gene. 56 strains were not sparfloxacin-resistance. There was not found to mutation of gyrA gene. However, the individual mutations of parC gene were accepted by 13 strains among 56 strains which showed sensitivity by levofloxacin. One strain was Asp-78->Asn, other one strain was Ser-79->Phe, and 11 strains were Lys-137->Asn. These results suggest that fluoroquinolone-resistance could be due to the multiple mutations in gyrA gene and parC gene, although the individual mutation of parC gene existed also in levofloxacin-susceptible strains.

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In order to study the prevalence of intestinal parasites among Japanese expatriates of developing countries, we conducted fecal specimen examinations annually from 1995 to 2000. Fecal specimens were collected from Japanese expatriates of each area: Asia, the Middle East, East Europe, Africa and Latin America. The specimens were fixed with 10% formalin in the local area, and then examined in Japan. We used the formalin-ether sedimentation method to find protozoan cysts or helminth eggs.
In 1995, the infection ratio was 3.0% (N=981), decreasing to 2.4% (N=1, 275) in 1996, 2.3% (N=1, 620) in 1997 and 1.6% (N=1, 574) in 1998. However, the ratio began to increase in 1999 (2.0%, N=1, 713) and 2000 (2.5%, N=1, 806). The ratio in Africa was the highest in each year, followed by Asia and Latin America. Heterophyidae (51 cases), Giardia lamblia (42) and Trichuris trichiura (30) were detected most frequently. In Egypt, the ratio of Heterophyidae rose by 28.8% in 2000. This was the prime reason for the increase in the overall ratio of infections worldwide. Most of the Japanese infected with Heterophyidae in Egypt had ingested dried mullet roe. This may explain the reason for the increase in the infection ratio there.
Although the prevalence of intestinal parasites among Japanese expatriates in developing countries is decreasing, those who eat fish in these areas are still at risk. In order to eradicate intestinal parasitosis from this group, we must continue preventive measures such as health education.

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The molecular identification of Mycobacterium tuberculosis DNA in ancient human remains has been achieved mainly in mummies with macroscopic changes but not in the skeletons without bone tuberculosis. Using polymerase chain reaction studies, we identified mycobacterial DNA in 2000-year-old human skeletons without pathological changes. Our findings suggest that these people suffered from an outbreak of tuberculosis. Molecular examinations for mycobacterial DNA in the bone marrow of skeletons may contribute to the clarification of ancient diseases in old human populations.

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We reviewed the clinical manifestations, sequential changes in cryptococcal antigen titers in serum and cerebrospinal fluid (CSF), and the antifungal drug susceptibility of Cryptococcus neoformans in three patients with cryptococcal meningitis between 1996 and 2000. Cryptococcal antigen titers were measured using the latex agglutination method with Pastrex Cryptococcus (Fuji Mebio, Tokyo) and Serodirect Cryptococcus (Eiken Chemical, Tokyo). The underlying systemic diseases in the three patients were liver cirrhosis, non-Hodgkin's lymphoma associated with miliary tuberculosis, and malignant thymoma associated with systemic lupus erythymatosus. The CSF samples showed positive indian ink staining in two of the three patients and C. neoformans was cultured from all three. The cryptococcal antigen titers in serum were higher than those in the CSF. The serum and CSF cryptococcal antigen titers measured by Serodirect Cryptococcus were higher than those measured by Pastrex Cryptococcus. The maximum titers of antigen in serum and CSF measured by Serodirect Cryptococcus were greater than 1, 024 in all three patients. The treatment regimens used for the three patients were amphotericin-B (AMPH-B) and flucytosine (5-FC), fluconazole (FLCZ) and intrathecal AMPH-B, FLCZ and 5-FC, and intrathecal AMPH-B, respectively. The antigen titers in serum and CSF decreased after treatment in all three patients. The antigen titers decreased slowly over 7.3 months in the most seriously ill patient who had non-Hodgkin's lyfnphoma associated with miliary tuberculosis. The time between the beginning of treatment and CSF cryptococal antigen titers falling to less than 8 was 1.7 to 7.3 months in the three patients, but the serum titers did not decrease to less than 8 during this period. The minimum inhibitory concentration was 0.06-0.25μg/ml for AMPH-B, 4-8μg/ml for 5-FC, 2-8μg/ml for FLCZ, 0.125-0.5μg/ml for miconazole and 0.03-0.125μg/ml for itraconazole. The measurement of sequential changes in cryptococcal antigen titers in serum and CSF was useful for evaluating the response to treatment.

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There were 71 patients with candidemia in our hospital from November 1, 1993 to October 31, 1 999. We investigated the 59 patients from isolated species, route of infection, underlying disorders, risk factors, complications, treatment and prognosis.Candida albicans was the most commonly isolated species (52%), followed by Candida tropicalis (11%). Eighty eight percent of the patients developed candidemia from central venous catheter related infections. The risk factors to candidemia included keeping the catheter in place for more than 5 days, gastrointestinal tract malignancies, postoperative state of gastrointestinal tract surgery, administration of broadspectrum or combination antibiotics for more than 5 days, and under corticosteroid therapy. About half of the patients (47%) had complications, including endophthalmitis (19 patients, 32%), septic shock (12 patients, 20%). Mortality rate associated with candidemia was 46%. Mortality rate was lower in 20 patients who were treated with amphotericin B (40%) than in 34 patients treated with only fluconazole (50%), but it was not statistically significant. In order to make an early diagnosis of candidemia, taking blood cultures and ophthalmologic examinations are essential, especially for patients who have those risk factors to candidemia mentioned above. If the patient was suspected of having catheter related infection, the catheter should be removed quickly and the catheter tip should be cultured. Once candidemia is found, ophthalmologic examination and systemic antifungal therapy are needed. Antifungal therapy with Amphotericin B should be used for patients with severe candidemia or with candidemia of non-albicans Candida species.

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After eating raw shellfish, 62-year-old man suffered from watery diarrhea and leg pain. The disease developed to septicemia and necrotizing fasiitis of bilateral lower extremities. Vibrio vulnificus was isolated from wound. He was treated with antibiotics, surgical debridement and hyperbaric oxygen therapy, and consequently was saved. Effect of hyperbaric oxygen therapy on V. vulnificus infection was discussed.

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We report a case of septicemia type Vibrio vulnificus infection. The patient was a 74-year-old man who had liver cirrhosis and hepatocelluler carcinoma. He felt a pain in the right femoral lesion after eating raw shellfish (Japanese “Umitake”) two days ago. He was admitted to our emergency center due to his shock status and thrombocytopenia two days after the onset. We diagnosed necrotizing fasciitis due to Vibrio vulnificus infection, his life was saved by emergency amputation of the right lower extremity. The culture of the blood and vesicle fluid showed Vibrio vulnificus. There are some reports that the debridement was effective to necrotizing fasciitis due to Vibrio vulnificus infection, but these reports are all about single upper extremity lesion. As far as we know, this is the second report of lower extremity necrotiaong fasciitis due to septicemia type Vibrio vulnificus infection rescued by extremity amputation in Japan. The mortality of septicemia type Vibrio vulnificus infection with necrotizing fasciitis is very high, this is quite a valuable report in making a decision for therapy of septicemia type Vibrio vulnificus infection.

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