A serious disease of amphibians caused by the chytrid fungus Batrachochytrium dendrobatidis was first found in Japan in December 2006 in imported pet frogs. This was the first report of chytridiomycosis in Asia. To assess the risk of pandemic chytridiomycosis to Japanese frogs, we surveyed the distribution of the fungus among captive and wild frog populations. We established a nested PCR assay that uses two pairs of PCR primers to amplify the internal transcribed spacer (ITS) region of a ribosomal RNA cassette to detect mild fungal infections from as little as 0.001 pg (1 fg) of B. dendrobatidis DNA. We collected swab samples from 265 amphibians sold at pet shops, 294 bred at institutes and 2103 collected at field sites from northern to southwestern Japan. We detected infections in native and exotic species, both in captivity and in the field. Sequencing of PCR products revealed 26 haplotypes of the B. dendrobatidis ITS region. Phylogenetic analysis showed that three of these haplotypes were specific to the Japanese giant salamander (Andrias japonicus) and appeared to have established a commensal relationship with this native amphibian. Many other haplotypes were carried by alien amphibians. The highest genetic diversity of B. dendrobatidis was found in the American bullfrog (Rana catesbeiana). Some strains of B. dendrobatidis appeared to be endemic to Japanese native amphibians, but many alien strains are being introduced into Japan via imported amphibians. To improve chytridiomycosis risk management, we must consider the risk of B. dendrobatidis changing hosts as a result of anthropogenic disturbance of the host-specific distribution of the fungus.

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We investigated whether manganese peroxidase (MnP) and the laccase-mediator system with 1-hydroxybenzotriazole (HBT) as mediator can remove the estrogenic activities of the steroidal hormones 17beta-estradiol (E(2)) and ethinylestradiol (EE(2)). Using the yeast two-hybrid assay system, we confirmed that the estrogenic activities of E(2) and EE(2) are much higher than those of bisphenol A and nonylphenol. Greater than 80% of the estrogenic activities of E(2) and EE(2) were removed following 1-h treatment with MnP or the laccase-HBT system; extending the treatment time to 8h removed the remaining estrogenic activity of both steroidal hormones. HPLC analysis demonstrated that E(2) and EE(2) had disappeared almost completely in the reaction mixture after a 1-h treatment. These results strongly suggest that these ligninolytic enzymes are effective in removing the estrogenic activities of E(2) and EE(2).

The present findings demonstrate that pioglitazone improves LV diastolic function without LV mass regression in hypertensive patients in proportion to the amelioration of insulin resistance. These findings suggest that increased adiponectin and MMP may be involved in the beneficial effect of pioglitazone on diastolic function.

Differences in susceptibility of five cladocerans to the neonicotinoid imidacloprid and the phenyl-pyrazole fipronil, which have been dominantly used in rice fields of Japan in recent years, were examined based on short-term (48-h), semi-static acute immobilization exposure tests. Additionally, we compared the species sensitivity distribution (SSD) patterns of both insecticides between two sets of species: the five tested cladocerans and all other aquatic organisms tested so far, using data from the ECOTOX database of U.S. Environmental Protection Agency (USEPA). The sensitivity of the test species to either imidacloprid or fipronil was consistent, spanning similar orders of magnitude (100 times). At the genus level, sensitivities to both insecticides were in the following descending order: Ceriodaphnia > Moina > Daphnia. A positive relationship was found between body lengths of each species and the acute toxicity (EC(50)) of the insecticides, in particular fipronil. Differences in SSD patterns of imidacloprid were found between the species groups compared, indicating that test cladocerans are much less susceptible than other aquatic species including amphibians, crustaceans, fish, insects, mollusks and worms. However, the SSD patterns for fipronil indicate no difference in sensitivity between cladocerans tested and other aquatic organisms despite the greater exposure, which overestimates the results, of our semi-static tests. From these results, Ceriodaphnia sp. should be considered as more sensitive bioindicators (instead of the standard Daphnia magna) for ecotoxicological assessments of aquatic ecosystems. In addition, we propose that ecotoxicity data associated with differences in susceptibility among species should be investigated whenever pesticides have different physicochemical properties and mode of action.