As the foods are stocked below freezing and widely distributed, a kind of food-borne outbreak which occurs in separate regions or in different time, so called “diffuse outbreak”, has been found at the present day. Unless the outbreak is early recognized, the number of victims would increase. Some methods have been developed to analyze the relatedness of bacteria isolated from the patients of enteric infections. PFGE, pulsed-field gel electrophoresis, is one of the methods and powerful to discriminate the difference in nucleotide sequences among bacterial genomes. Availability of PFGE analysis is appreciated to examine the linkage of each incident of food-borne infections in epidemiological investigation. A PFGE network, PulseNet Japan, is now under construction among National Institute of Infectious Diseases, local Health Institutes and Ministry of Health, Labour and Welfare.

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We identified the species or genotypes of the six Cryptosporidium isolates from patients and C. parvum strain HNJ-1 using the seven previously described species-differentiation and genotyping PCR protocols for detection of Cryptosporidium parasites. In addition, we also discussed about the usefulness of these PCR-based protocols on the basis of the reports previously published.
Cryptosporidium diagnostic fragment was amplified by PCR with each primer pair, targeting the 18S ribosomal RNA (18SrRNA), Cryptosporidium oocyst wall protein (COWP), Heat shock protein 70 (HSP70), Polythreonine (Poly-T), Thrombospondin related adhesive protein of Cryptosporidium-1 (TRAP-C1), and unknown gene locus, in all isolates from patients and the strain HNJ-1. The RFLP profiles of 18SrRNA, COWP, HSP70, Poly-T, and TRAP-C1 PCR products in all isolates from patients were found to be the same among isolates, and were correspondent to those of C. parvum human genotype. While the RFLP profiles of HNJ-1 were strictly different from those of isolates from patients, and were correspondent to C. parvum cattle genotype. In addition, nucleotide sequences in 18 SrRNA gene of all isolates from patients and HNJ-1 were found to be identical to that of C. parvum human or cattle genotype, respectively. Therefore, the isolates from patients and HNJ-1 were identified as C. parvum human and cattle genotype, respectively.
According to the reports related to the PCR-based protocols applied in the present study, RFLP profiles targeting the HSP70, Poly-T, TRAP-C1 genes had been revealed in only a few species or genotypes, but those of 18SrRNA and COWP genes were in all species and genotypes. However, we supposed that it was difficult to distinguish between human or cattle genotype and other species or genotypes by RFLP profiles of 18SrRNA or COWP because the RFLP profiles of human or cattle genotype were identical or similar to those of other species or genotypes. On the other hand, it has been known that the nucleotide sequences in 18SrRNA or COWP gene are different among Cryptosporidium species and/or genotypes. Therefore, the direct sequencing method targeting the variable regions which can be used to distinguish among Cryptosporidium species, as well as the genotypes within C. parvum in either 18SrRNA or COWP gene is the most useful tool for accurate identification of Cryptosporidium isolates.

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Recently travelers who were bitten by possibly rabid animals in rabies endemic regions and returned to Japan have increased in number. About half of them received rabies post-exposure prophylaxis (RPEP) with one or more doses of foreign-made rabies vaccines (FRV) in the local medical institutions. FRV, however, are not available in Japan so we have to continue the RPEP with Japanese rabies vaccine (JRV). It has not been demonstrated that an anti-rabies antibody induced with JRV following Vero cell rabies vaccine (PVRV) or chick embryo cell rabies vaccine (PCEC) could be high enough to prevent clinical rabies. We examined anti-rabies antibody (ARA) titers among the subjects visited our vaccine clinic to receive RPEP and obtained results as follows: the ARA titers after a total of 5 doses of PCEC or PVRV and JRV were high enough to prevent clinical rabies as after 5 doses of JRV. However, ARA titers obtained after receiving one dose of PVRV and 2 doses of JRV seemed lower than those produced after one dose of PCEC and 2 doses of JRV or 3 doses of JRV. To accelerate antibody production, consequently, the simultaneous intradermal and subcutaneous injection method of rabies vaccine may be applied to those who were bitten in their hands or head by possibly rabid animals and received only one dose of PVRV in rabies endemic regions.

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The case was a 28-year-old Japanese female who was considered to be infected with malaria in India. She manifested fever in Tokyo, Japan, and was brought to Toho University Hospital due to continuous high fever and severe thrombocytopenia. Ring forms at 11% of her RBCs and ICT^TM Malaria P. f/P. v test was also positive for Plasmodium falciparum diagnosis. Not only the high parasitemia and delay of the diagnosis (6 days after the onset of fever), but also her DIC status required prompt and proper treatment. The diagnosis of severe malaria was strongly considered, and intravenous Artesunate (a qinghaosu derivative) was decided to be administered to the patient. After the four series of administration, mefloquine was subsequently given to prevent recrudescence. Parasite clearance time and fever clearance time were 24 hours and 108 hours, respectively. Thrombocytopenia was improved shortly after the treatment, but then anemia was once worsened with following gradual improvement. No other significant side effects were observed and no recrudescence occurred up to 8 months after her discharge. In Japan, very few cases treated with intravenous Artesunate were reported and our results showed its safe and excellent effect for a Japanese malaria patient.

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A 46-year-old man refer to us because of hemospermia. The prostatic gland was normal in size and consistency at rectal examination. Serum prostate specific antigen was 7.04ng/ml. Magnetic resonance imaging showed an area of low signal intensity on T2-weighted images in the left peripheral gland, possibly indicative of carcinoma. Transrectal prostate biopsy was performed after intravenous administration of piperacillin. He developed chills and fever (39°C) the next morning following biopsy. He was taken unconscious into the hospital where a diagnosis of septic shock caused by Escherichia coli was made. Five days later, he died. His general condition deteriorated notwithstanding intensive treatment. Postmortem blood cultures were positive for a piperacillin resistant Escherichia coli. Histological examination of the biopsies showed a benign prostatic hyperplasia. Autopsy showed diffuse tissue damage in the heart, lung, liver and kidneys. The prostate had numerous microabscesses.
Currently, transrectal prostate biopsy is considered a generally reliable procedure to detect adenocarcinoma of the prostate. Our case seems to the sixth case report of fatal complications.

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