Assays of dioxins and dioxin-like compounds in actually contaminated soils using transgenic tobacco plants carrying a recombinant mouse aryl hydrocarbon receptor-mediated β-glucuronidase reporter gene expression system

The transgenic tobacco plant XD4V-26 carrying the recombinant mouse aryl hydrocarbon receptor XD4V-mediated β-glucuronidase (GUS) reporter gene expression system was used for assay of dioxins and dioxin-like compounds consisting of polychlorinated dibenzeno-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls (Co-PCBs) in actually contaminated soils. The transgenic tobacco plant XD4V-26 showed a significant dose-dependent induced GUS activity when cultured on MS medium containing PCB126 [toxic equivalency factor (TEF) = 0.1]. In contrast, PCB169 and PCB180, which have 0.03 of TEF and unassigned TEF values, respectively, did not significantly induce GUS activity under the same conditions as with PCB126. When the tobacco plants were cultivated for up to 5 weeks on actually contaminated soils with dioxins and dioxin-like compounds collected from the periphery of an incinerator used for disposal of residential and industrial wastes, GUS activity in the leaves was dose-dependently increased. The plants clearly detected 360 pg-TEQ g(-1) of dioxins and dioxin-like compounds in this assay. There was a positive correlation between GUS activity and TEQ value of dioxins and dioxin-like compounds in the plants. This assay does not require any extraction and purification processes for the actually contaminated soil samples.     

Novel enzymatic activity of cell free extract from thermophilic Geobacillus sp. UZO 3 catalyzes reductive cleavage of diaryl ether bonds of 2,7-dichlorodibenzo-p-dioxin

We characterized the ability of the cell free extract from polychlorinated dibenzo-p-dioxins degrading bacterium Geobacillus sp. UZO 3 to reduce even highly chlorinated dibenzo-p-dioxins such as octachlorodibenzo-p-dioxins in incineration fly ash. The degradation of 2,7-dichlorodibenzo-p-dioxin (2,7-DCDD) as a model dioxin catalyzed by the cell free extract from this strain implicates that the ether bonds of 2,7-DCDD molecule undergo reductive cleavage, since 4′,5-dichloro-2-hydroxydiphenyl ether and 4-chlorophenol were detected as intermediate products of 2,7-DCDD degradation.     

Formation of chlorinated estrones via hypochlorous disinfection of wastewater effluent containing estrone

Chlorinated derivatives of estrone (E1) in the effluent of a municipal sewage treatment plant located in Shizuoka prefecture, Japan were detected by gas chromatography/mass spectrometry using electron impact in selected ion monitoring (GC/MS-EI-SIM) analysis. The concentrations of E1, 2-chloroestrone, 4-chloroestrone and 2,4-dichloroestrone in the effluent sample collected in December 2005 were 60.0 ng l(-1), 4.0 ng l(-1), 14.5 ng l(-1), and 9.8 ng l(-1), respectively. In the effluent sample taken in June 2005, 2,4-dichloroestrone was detected at 5.6 ng l(-1) along with 17.6 ng l(-1) of E1. However, only E1 was detected at 5.9 ng l(-1) in the sample in May 2005. To elucidate the behavior of E1 during the disinfection process with sodium hypochlorite in the sewage treatment plant, we carried out a reaction of E1 with sodium hypochlorite in buffer solutions at pH 7 and 9. As E1 was consumed rapidly, chlorinated estrones were produced and relatively fast formation of 2-chloroestrone, 4-chloroestrone, and 2,4-dichloroestrone was observed. Furthermore, 1,4-estradiene-3,17-dione derivatives were formed from the reaction between 2,4-dichloroestrone and sodium hypochlorite.     

Dioxins in human milk and smoking of mothers

BACKGROUND: The relation between the levels of dioxins in human breast milk and the smoking habits of the mothers is controversial. To clarify this relationship, we analyzed data from the human milk survey in Japan. METHODS: The human milk survey has been conducted in Japan since 1997. Healthy pregnant women aged 20-39 years were recruited and 50 ml of breast milk was collected from them at 30 days after delivery. PCDDs, PCDFs, and dioxin-like PCBs were measured by using GC/MS. The smoking habits of the mothers were established by interviewing them soon after delivery and were classified into four categories: current smokers, ever smokers who quit smoking at the pregnancy, ever smokers who quit smoking before the pregnancy, and never smokers. The levels of dioxins in breast milk were compared in the four categories of smoking among 853 primiparas. In addition, we analyzed the association between dioxin levels and passive smoking among never smokers. The geometric means of the dioxin concentrations were calculated in order to compare the differences between dioxins. RESULTS: The geometric means of dioxin-like PCBs in milk of never smokers was the highest (9.2 pg TEQ/g fat); followed by ever smokers who quit smoking before the pregnancy, ever smokers who quit smoking at the pregnancy, and current smokers (7.5, 7.2, and 6.6 pg TEQ/g fat, respectively). The differences between these levels were statistically significant (ANOVA, p<0.001). No significant difference was observed between the level of dioxins in milk from never smokers subjected to passive smoking status and those who had not experienced passive smoking. CONCLUSION: The levels of dioxin-like PCBs in human milk were negatively related to the smoking habits of mothers.     

Comparative fertilizer properties of digestates from mesophilic and thermophilic anaerobic digestion of dairy manure: focusing on plant growth promoting bacteria (PGPB) and environmental risk

The fertilizer properties of anaerobic digestate depend on the feedstock and operating conditions of digestion. In this study, the comparative fertilizer properties of mesophilic and thermophilic digestates from dairy manure were evaluated for plant nutrient contents, and special attention was paid to plant growth promoting bacteria (PGPB). Two digestates contained similar plant nutrient contents, while the thermophilic digestate contained higher contents of NH₄⁺–N. The quantity of Bacillus and Pseudomonas in the mesophilic digestate was significantly higher than in the thermophilic digestate. Furthermore, Bacillus showed siderophore production and antifungal activity (43.5–75.3%), and Pseudomonas showed siderophore and phytohormone production (4.2–75.2 µg ml^?1). One phosphate solubilizing isolate was also detected in the mesophilic digestate. These results indicated that two digestates showed different fertilizer properties with respect to nutrient contents and PGPB, and digestates had the potential to increase the availability of phosphorus and iron in the soil, both to provide phytohormones to plant roots and protect plants from fungal phytopathogens. The contents of indicator bacteria and heavy metals were analyzed to determine their environmental risk, and the results showed a high reduction in indicator bacteria and lower levels of heavy metals than in other feedstocks.     

Lysis of cyanobacteria with volatile organic compounds

One of bacteria collected from Lake Sagami, Japan, Brevibacillus sp., was found to have a lytic activity of cyanobacteria, but did not produce active compounds. Instead, the co-culturing of Microcystis with the Brevibacillus sp. enhanced the production of two volatile compounds, beta-cyclocitral and 3-methyl-1-butanol, and the former had a characteristic lytic activity. It was confirmed that these volatile compounds were derived from the cyanobacteria themselves. beta-Ionone, geosmin and 2-methylisoborneol derived from cyanobacteria and similar volatile compounds, terpenoids, produced by plants also had a lytic activity. The minimum inhibitory concentration values of the cyanobacterial metabolites were estimated to be higher than those of compounds from plants except for a few compounds. Among them, beta-cyclocitral only produced a characteristic color change of culture broth from green to blue. This color change is similar to the phenomenon observed when a sudden decline in growth of cyanobacteria begins in a natural environment.     

Geology-dependent impacts of forest conversion on stream fish diversity

Forest conversion is one of the greatest global threats to biodiversity, and land-use change and subsequent biodiversity declines sometimes occur over a variety of underlying geologies. However, how forest conversion and underlying geology interact to alter biodiversity is underappreciated, although spatial variability in geology is considered an integral part of sustaining ecosystems. We aimed to examine the effects of forest conversion to farmland, the underlying geology, and their interaction on the stream fishes’ diversity, evenness, and abundance in northeastern Japan. We disentangled complex pathways between abiotic and biotic factors with structural equation modeling. Species diversity of stream fishes was indirectly shaped by the interaction of land use and underlying geology. Diversity declined due to nutrient enrichment associated with farmlands, which was mainly the result of changes in evenness rather than by changes in species richness. This impact was strongest in streams with volcanic geology with coarse substrates probably because of the differential responses of abundant stream fishes to nutrient enrichment (i.e., dominance) and the high dependency of these fishes on large streambed materials during their life cycles. Our findings suggest that remediation of deforested or degraded forest landscapes would be more efficient if the interaction between land use and underlying geology was considered. For example, the negative impacts of farmland on evenness were larger in streams with volcanic geology than in other stream types, suggesting that riparian forest restoration along such streams would efficiently provide restoration benefits to stream fishes. Our results also suggest that land clearing around such streams should be avoided to conserve species evenness of stream fishes.     

Distribution of Todarodes pacificus (Cephalopoda: Ommastrephidae) paralarvae near the Kuroshio off southern Kyushu, Japan

The distribution and abundance of Japanese common squid [Todarodes pacificus (Steenstrup, 1880)] paralarvae off southern Kyushu Island near the Kuroshio were examined in relation to water types in the region. Surveys were conducted in 1996 and 1997 using paired, 70 cm diameter Bongo nets. Temperature–salinity plots from 0, 50 and 100 m depths were used to assign sampling stations to three water types: inshore, mixing and Kuroshio. In total, 4103 T. pacificus paralarvae were collected at 59 of 72 stations. Catch densities at positive stations ranged between 8 and 4282 ind./50 m² sea surface area. Catches during both years were highest in mixing waters, followed by inshore and Kuroshio waters. Total catches were highest just north of the oceanographic front at the inshore edge of the Kuroshio and declined with increasing distance away from the front. Catches of paralarvae ≤1.0 mm mantle length also peaked near the front. In both years, ≥85% of all paralarvae and ≥69% of those ≤1.0 mm mantle length occurred less than 25 km from the front. We conclude that most spawning off southwest Kyushu occurs near the frontal zone, where Kuroshio and inshore waters meet. Received: 18 November 1998 / Accepted: 11 May 1999     

Fluorinated alkyl compounds including long chain carboxylic acids in wild bird livers from Japan

A wide range of fluorinated alkyl compounds (FACs) has been reported in wildlife in various locations in the world. However, such information regarding Japanese wildlife is rarely found. In the present study, we investigated the occurrence of 21 FACs, including perfluorinated alkyl sulfonates (PFASs), perfluorinated carboxylates (PFCAs), and fluorotelomer acids, in the livers of 10 wild bird species from two regions in northern Japan. To avoid interferences, FACs were quantified by a recently developed method using acetonitrile and solid-phase extraction followed by an ion exchange HPLC column separation. Apart from perfluorooctane sulfonate (PFOS), which was found at the highest levels of all the compounds detected, several long chain perfluorinated carboxylates (PFCAs) from C8 to C16, particularly perfluorotetradecanoic acid (PFTeDA) and perfluorohexadecanoic acid (PFHxDA), were detected for the first time. Additionally, 7:3 FTCA, a fluorotelomer acid, was also detected in most swan livers from Miyagi prefecture and all the birds from Tochigi prefecture. However, none of the sulfonamides and unsaturated telomer acids were detected in any species. Swans seem to be the least exposed wild birds to FACs among the investigated birds, signifying that feeding habits may reflect FAC accumulation in wild birds. The highest total concentration of detected FACs was 405 ng g⁻¹ wet wt., which was found in a Japanese sparrowhawk, indicating that the top predatory wild birds can accumulate several long chain carboxylic acids. However, the current FAC concentrations found in livers may suggest that these compounds alone would not cause a severe toxic effect in these species.     

Organochlorine concentrations in breast milk and prevalence of allergic disorders in Japanese women

Persistent organic pollutants have been shown to have immunomodulating effects in humans. However, epidemiological evidence regarding the relationships between organochlorine compound exposure and allergic disorders coming from studies of children has been limited and inconsistent. The current cross-sectional study examined the associations between the concentrations of β-hexachlorocyclohexane (HCH), hexachlorobenzene (HCB), p,p'-dichlorodiphenyldichloroethylene (DDE), and trans-nonachlordane in breast milk and the prevalence of allergic disorders in 124 adult Japanese women. The definition of wheeze and asthma was based on criteria from the European Community Respiratory Health Survey whereas that of eczema and rhinoconjunctivitis was based on criteria from the International Study of Asthma and Allergies in Childhood. Adjustment was made for age, smoking, family history of allergic disorders, and education. The prevalence values of wheeze, asthma, eczema, and rhinoconjunctivitis in the past 12 months were 9.7%, 4.8%, 13.7%, and 29.8%, respectively. The median concentrations of β-HCH, HCB, p,p'-DDE, and trans-nonachlordane in breast milk were 28.3, 7.0, 71.6, and 23.9 ng g(-1) lipid, respectively (range, 4.5-253, 2.1-14.5, 7.5-362, and 1.8-130 ng g(-1) lipid, respectively). When the exposures were treated as continuous variables, no significant associations were found between concentrations of HCB, β-HCH, p,p'-DDE, or trans-nonachlordane and the prevalence of wheeze, asthma, eczema, or rhinoconjunctivitis. Our results suggest that concentrations of β-HCH, HCB, p,p'-DDE, and trans-nonachlordane in breast milk are not evidently associated with the prevalence of wheeze, asthma, eczema, or rhinoconjunctivitis in young female Japanese adults.