Isolation and habitat analysis of Balamuthia mandrillaris from soil.

Balamuthia mandrillaris is a free-living amoeba that causes meningoencephalitis in mammals. Over 200 cases of infection were reported worldwide, with a fatality rate of over 95%. A clear route of infection was unknown for a long time until a girl died of granulomatous amoebic encephalitis (GAE) in California, USA, in 2003 due to infection with B. mandrillaris detected in a potted plant. Since then, epidemiological studies were conducted worldwide to detect B. mandrillaris in soil and other environmental samples. We previously reported the isolation of B. mandrillaris from the soil in Japan; however, the existing B. mandrillaris culture method with BM3 medium and COS-7 cells was unsuccessful. Therefore, in this study, we aimed to conduct soil analysis to determine the growth conditions of B. mandrillaris. B. mandrillaris-positive soils were defined as soils from which B. mandrillaris was isolated and environmental DNA was PCR-positive. Soils inhabited by B. mandrillaris were alkaline, with high electrical conductivity and characteristics of nutrient-rich soils of loam and clay loam. The results of this study suggest a possible reason for the high prevalence of GAE caused by B. mandrillaris among individuals employed in agriculture-related occupations.

DETERMINATION OF GUT BACTERIAL METABOLITES IN RADIATION EXPOSED MICE.

Gut microflora (GM) impacts human health in various ways, both beneficial and detrimental. Recently, it has attracted attention for its application in treatment, as protective agents, and as biomarkers in radiation exposure. In this study, we focused on organic acids that have not yet been reported to be related to radiation exposure; we measured the pH and organic acid content in the faeces of 0, 2, 4 and 8 Gy-irradiated mice. A common trend of fluctuation of some organic acids was observed in each group, suggesting a correlation between radiation exposure and organic acid fluctuation. Lactate fluctuation was similar between 0 and 2 Gy-, and 4 and 8 Gy-irradiated mice. Based on this finding, we suggest that lactate may also be an organic acid that is greatly affected by irradiation.

Short-term follow-up of intestinal flora in radiation-exposed mice.

Some gastrointestinal bacteria, otherwise known as the 'intestinal flora', can cause severe gastrointestinal problems, including sepsis, which are strongly linked to lifestyle-related diseases, including cardiovascular diseases. Several investigations have focused on the long-term changes in the intestinal flora associated with radiation exposure; however, the short-term effects remain unknown. In this study, we tracked the short-term changes in the intestinal flora of mice exposed to different doses of X-ray irradiation (2 Gy and 4 Gy), focusing only on the lactic acid bacteria Bifidobacterium and Lactobacillus. A decrease in the Lactobacillus abundance was detected immediately after irradiation in individuals exposed to both 2 Gy and 4 Gy irradiation. However, mice exposed to 4 Gy of irradiation showed a remarkable increase in Bifidobacterium, indicating a potential role of these bacteria in regeneration of the intestinal epithelial tissue. Studies on changes in intestinal bacteria as a result of radiation exposure are limited. Therefore, continuation of this field of research is expected to provide important fundamental insight into the mechanisms by which radiation causes damage to the intestinal tissues, contributing to the development of sepsis.

First report of the isolation of Balamuthia mandrillaris in the northern region of Japan.

Balamuthia mandrillaris is a free-living amoeba that lives in soil and water near human settlements. B. mandrillaris was first isolated from a mandrill baboon that died at the San Diego Zoo Wildlife Park in California in 1986, and the first human infection was reported in 1990. Although reported B. mandrillaris infections are often not properly characterized, it appears that B. mandrillaris invades the living body from the soil and water, either via a wound or the nasal cavity. Most confirmed infections have originated in South and North America. B. mandrillaris inhabits warm climates and is recognized as a pathogen in warm areas such as desert climates and tropical climates. B. mandrillaris has been isolated from environmental samples since 2000, most of which originated from warm areas such as step climates, tropical climates, and desert climates. However, B. mandrillaris may survive in diverse environments, although fewer granulomatous amebic encephalitis patients have been reported in colder Japanese and Northern European regions. In the present study, we conducted a survey of 13 soil samples in Aomori Prefecture located at the northernmost tip of Japan Honshu and successfully isolated one strain of B. mandrillaris from soil for the first time in Japan. In addition, B. mandrillaris gene was detected from several soils. This confirms that B. mandrillaris is capable of spreading to a wider climatic region.

Expression of metallothionein mRNAs on mouse cerebellum microglia cells by thimerosal and its metabolites.

Effects of thimerosal and its metabolites, ethyl mercury and thiosalicylate, on the expression of metallothionein (MT) mRNAs in mouse cerebellum microglia cell line, C8-B4 cells, were studied. The level of MT-1 mRNA significantly decreased at early hours and recovered time-dependently 24h after thimerosal was added to the C8-B4 cells. However, MT-2 and MT-3 mRNA expressions did not change from the control group. In contrast, the expression of MT-1 mRNA increased in a mouse neuroblastoma cell line 6h after incubation with thimerosal. In addition, the level of MT-1 mRNA decreased in C8-B4 cells 6h after the addition of thiosalicylate, but ethyl mercury induced MT-1 mRNA expression. When cell viability was compared with thimerosal, thiosalicylate, and ethyl mercury, the viability of C8-B4 cells decreased dose-dependently 24h after either thimerosal or ethyl mercury was added; however, the viability increased dose-dependently until 15 microM thiosalicylate was added. From the present results, it is concluded that the expression of MT-1 mRNA may be mediated by different factors than the expression of MT-2 mRNA in C8-B4 cells. The reduction of MT-1 mRNA level by thiosalicylate may affect the proliferation of C8-B4 cells.