Effects of heavy metals on sea urchin embryo development. Part 2. Interactive toxic effects of heavy metals in synthetic mine effluents

Interactive toxic effects between heavy metals were investigated using a sea urchin (Anthocidaris crassispina) bioassay. An effluent from an abandoned mine showed significant inhibitory effects on embryo development as well as producing specific malformations. The effects on the embryos were reproduced by synthetic polluted seawater consisting of eight metals (manganese, lead, cadmium, nickel, zinc, chromium, iron, and copper) at the concentrations detected in the mine effluent. This indicated that the heavy metals were responsible for the effects observed. Five heavy metals were ranked in decreasing order of toxicity as follows: Cu>Zn>Pb>Fe>Mn. Among these, zinc and manganese could cause malformation of the embryos. From bioassay results using 27 combinations of heavy metals, 16 combinations including zinc could produce specific malformations, such as radialized, exo-gastrulal, and spaceship Apollo-like gastrulal embryos. Zinc was one of the elements responsible for causing malformations and its effects were intensified by the presence of the other metals, such as manganese, lead, iron, and copper.     

Variation in CO2 assimilation rate induced by simulated dew waters with different sources of hydroxyl radical (*OH) on the needle surfaces of Japanese red pine (Pinus densifora Sieb. et Zucc.)

The hydroxyl radical (*OH) is generated in polluted dew on the needle surfaces of Japanese red pine (Pinus densiflora Sieb. et Zucc.). This free radical, which is a potent oxidant, is assumed to be a cause of ecophysiological disorders of declining trees on the urban-facing side of Mt. Gokurakuji, western Japan. Mists of *OH-generating N(III) (HNO2 and NO2-) and HOOH + Fe + oxalate solutions (50 and 100 microM, pH 5.1-5.4) simulating the dew water were applied to the foliage of pine seedlings grown in open-top chambers in the early morning. Needles treated with 100 microM N(III) tended to have a greater maximum CO2 assimilation rate (Amax), a greater stomatal conductance (g(s)) and a greater needle nitrogen content (Nneedle), suggesting that N(III) mist acts as a fertilizer rather than as a phytotoxin. On the other hand, needles treated with 100 microM HOOH + Fe + oxalate solution showed the smallest Amax, g(s), and Nneedle, suggesting that the combination of HOOH + Fe + oxalate caused a decrease in needle productivity. The effects of HOOH + Fe + oxalate mist on pine needles were very similar to the symptoms of declining trees at Mt. Gokurakuji.     

Mutagenic nitrated benzo[a]pyrene derivatives in the reaction product of benzo[a]pyrene in NO2-air in the presence of O3 or under photoirradiation

In order to clarify the contribution of nitrated products to the direct-mutagenic activity of products of the reactions of benzo[a]pyrene in NO2-air under various conditions, heterogeneous reactions of BaP deposited on filter in the air containing 10 ppm of NO2 have been conducted in dark or under photoirradiation. The reaction products have been analyzed by gas chromatography and mutagenicity of the products fractionated by preparative HPLC was assayed for Salmonella typhimurium strains TA98 and YG1024 in the absence of S9 mix. 3,6-dinitrobenzo[a]pyrene and 1,3-dinitrobenzo[a]pyrene, which are strong direct-acting mutagens, largely contributed to the total direct-acting mutagenicity of the dark reaction products in NO2-air. On the other hand, both the dark reaction in the presence of O3 and the photoreaction in NO2-air resulted in the formation of much smaller amounts of nitrobenzo[a]pyrenes than that observed in the dark reaction in the absence of O3. These results show that the contribution of other direct-acting mutagens to the total direct-acting mutagenicity of the products in these reactions should be considered. Benzo[a]pyrene lactones were identified in a highly mutagenic fraction of the products of the dark reaction in the presence of O3 and photoreaction and a nitrobenzo[a]pyrene lactone was also identified in a highly mutagenic fraction of the dark reaction products in the presence of O3. Nitrated oxygenated benzo[a]pyrene derivatives such as nitrobenzo[a]pyrene lactone were considered to largely contribute to direct-acting mutagenicity of the products of the dark reaction in the presence of O3 and photoreaction.     

Application of lean flammability limit study and large eddy simulation to burner development for an oxy-fuel combustion system

Previously, we developed a model to predict lean flammability limit L and flame propagation velocity Sb for pulverized coal. In the present paper, we have extended the model to apply it in development of oxy-fuel combustion systems. The basic model consists of two particles. One of the two particles burns first, then, the other particle is ignited by the heat of combustion of the one burning particle. We analyzed at what distance the first burning particle could ignite the next particle, and how fast the first burning particle could ignite the next particle. The model was verified both for air and oxy-fuel combustion conditions. Next, a method to support burner development was examined by using the model. Local Sb and L near the ignition points of the burner could be analyzed from the concentration and temperature profiles of CFD results. Flame stability was judged by the calculated Sb and L profiles, and past results of blow-off limits obtained with actual- and pilot-scale experiments. A DS^®T-burner was developed by Hitachi Power Europe, and installed at the Schwarze Pumpe pilot plant. Combination of the technique and large eddy simulation was applied to confirmation of the system.     

Accumulation of cadmium and zinc in <Emphasis Type="Italic">Evodiopanax innovans</Emphasis>

The use of tree species for phytoremediation of contaminated soil offers the advantage of a large biomass in which to store contaminants. We investigated the cadmium (Cd) and zinc (Zn) accumulation ability of Evodiopanax innovans, a common deciduous tree species belonging to the Araliaceae family and widely found in secondary forests in Japan. Sampling was conducted at an old silver mine. Leaf samples were collected from nine tree species, including E. innovans. The seasonal variation of metal concentrations in the leaves and the detailed distribution of metals in the leaves and twigs of E. innovans were measured. We also analyzed the contents of organic acids in the leaves. The highest concentration of Cd in the leaves of E. innovans was 118 μg/g, which exceeds the threshold level for being considered a Cd hyperaccumulator (100 μg/g). For Zn, the highest value was 1040 μg/g in leaves, which is less than required to qualify as a Zn hyperaccumulator. Both Cd and Zn were found to accumulate in the petioles and veins of leaves and the bark of twigs. Since the oxalic acid content of leaves showed a weak correlation with Cd concentration, oxalic acid may play an important role in the accumulation of Cd. Taking both the Cd concentration level and the biomass of this woody plant into consideration, it may be possible to use E. innovans for the phytoremediation of Cd-contaminated soils.     

Modelling of stem biomass accumulation in Pinus densiflora seedlings exposed to aqueous-phase OH radicals generating mist

The present study aimed to evaluate the short-term biomass accumulation of forest trees exposed to wet acidic depositions. A hierarchical Bayesian model of tree growth was developed based on the data of a short-term experiment in which 2-year-old Japanese red pine (Pinus densiflora Sieb. et Zucc.) seedlings were exposed to aqueous phase OH radicals generated by an iron-oxalate-H₂O₂ mist (a pseudo-polluted dew) over two growing periods. We conducted a statistical comparison of tree growth between the control and pollution treatment groups by using the growth model incorporated the random effects due to the unknown characteristics of each seedling. The variability among seedlings is expressed in this model by the posterior probabilistic distributions of unobserved dry weight of a stem cohort before exposure treatment. The analysis of the effects of pollution treatment on the stem growth revealed that this treatment decreases the biomass allocation in the current year stems. However, the effects on the relative growth rate of pre-existing stems were unclear. Based on these results, we can speculate that in a polluted environment, the short-term growth of the young stems in the seedlings inhibited by pollution treatment, thereby resulting in the slowdown of long-term biomass accumulation. This can explain the patterns observed in the declining Japanese red pine forests that are subjected to OH-generating dews in the polluted area of western Japan.     

Evaluation of an electrolysis apparatus for inactivating antineoplastics in clinical wastewater

We recently reported a system for inactivating antineoplastics in which sodium hypochlorite is supplied by the electrolysis of sodium chloride solution. In this study, we designed an electrolysis apparatus for inactivating the cytotoxicity of antineoplastics in clinical wastewater using the system. The apparatus consists of an electrolysis cell with platinum-iridium electrodes, a pool tank, a circulating system for wastewater, a safety system for explosive gas and overflow, and an exhaust duct. The free chlorine concentration increased linearly up to 6500 mg l(-1), and pH also increased to 9.0-10.0 within 2h, when 0.9% sodium chloride solution was electrolyzed. We examined its efficacy with model and clinical wastewaters. The reciprocal of dilution factor for disappearance of cytotoxicity using Molt-4 cells was compared before and after electrolysis. In the model wastewater, that was 9.10 x 10(4) before electrolysis, and 3.56 x 10(2) after 2h of electrolysis. In the clinical wastewater (n=26), that was 6.90 x 10(3)-1.02 x 10(6) before electrolysis, and 1.08 x 10(2)-1.45 x 10(4) after 2h of electrolysis. Poisonous and explosive gases released by the electrolysis were measured; however, they were found to be negligible in terms of safety. The environmental load was evaluated by carbon dioxide generation as an index and it was found that the carbon dioxide generated by the electrolysis method was 1/70 lower than that by the dilution method with tap water. Moreover, the cost of the electrolysis method was 1/170 lower than that of the dilution method. This method was found to be both effective and economically valuable.     

Suppression of sexual behavior in male Japanese medaka (Oryzias latipes) exposed to 17beta-estradiol

Mating pairs of Japanese medaka (Oryzias latipes) received 17beta-estradiol (E2; dose, 3 and 30 microg/g body weight daily) in their diet and E2-free diet (control) for two weeks. The effect on reproductive success was assessed in the mating pairs, and sexual and nonsexual behaviors in the male were quantified. The fecundities of the E2-treated groups were reduced significantly (p < 0.05) compared with those of control, and the sexual behaviors (following, dancing, floating, and crossing) of male fish in response to prostaglandin F2alpha (PG)-injected E2-unexposed females were suppressed dramatically. While the fertility of spawned eggs and the frequencies of the nonsexual behaviors of resting and swimming in a straight line performed by male were unchanged in E2-exposed groups. Therefore, sexual behavior in male may be linked with reproductive success in mating pair of madaka exposed to E2. This assay for sexual behavior using PG may provide a useful tool for assessing the effect of endocrine-disrupting chemicals.     

Levels of PCDDs, PCDFs and Co-PCBs in human milk in Saitama, Japan, and epidemiological research

From 1998 to 2000, the dioxin levels were measured in the milk of 299 mothers who lived in Saitama prefecture, Japan. Factors that influenced the dioxin levels were investigated based on a questionnaire given to the milk donors at that time. It was found that the dioxin levels in the milk of the mothers who smoked were lower than those of non-smokers, and the dioxin levels were generally low in the mothers who were heavy smokers. The average dioxin levels in the milk of mothers who had been breast-fed were higher than those given formula, and there was a significant difference in the dioxin congeners. It was also found that the dioxin levels in milk of the women who regularly consumed fish and shellfish were generally higher. There was a strong correlation between summation operator TEQ and PCB126. The data suggested that PCB126 could be a useful indicator for the simplified analysis of dioxin in human milk.     

Comparison of polybrominated diphenyl ethers in fish, vegetables, and meats and levels in human milk of nursing women in Japan

At present, little is known about the occurrence of polybrominated diphenyl ethers (PBDEs) in fish and food products sold in Japan. To investigate whether human exposure to PBDEs through the diet is significant, the concentrations of PBDEs were determined in fish and in meat and vegetables sold in two food markets in the city of Hirakata, Osaka prefecture. The concentrations of PBDEs in the breast milk of 12 primiparae nursing women at one month after delivery also were determined to ascertain the relationship between the levels found in the diet and levels in humans. sigma PBDE concentrations ranged between 21 and 1650 pg/g fresh weight in the edible tissues of five species of fish and one shellfish species. The highest concentrations were measured in yellow-fin tuna, followed by short-necked clam, salmon, yellowtail, mackerel and young yellowtail. Interestingly, sigma PBDE concentrations were not statistically significantly different in two cultured mackerel samples from Japan and mackerel collected from northern European waters. sigma PBDE concentrations in beef, pork and chicken meat (ranging between 6.25 and 63.6 pg/g fresh weight) and in three different vegetables (ranging between 38.4 and 134 pg/g fresh weight) were significantly lower than the concentrations in fish. In human milk, sigma PBDE concentrations ranged between 668 and 2840 pg/g lipid basis, which is comparable to the levels in populations of nursing women reported in Sweden and elsewhere. There was a strong positive relationship between PBDE concentrations in human milk and dietary intake of fish and shellfish, which was established in the women from responses to a questionaire on food consumption habits. The results of this study of food products commonly consumed in Japan and the levels found in nursing women raise a concern about low level PBDE contamination of fish and other foods intended for human consumption.