Oxidation state specific analysis of arsenic species in tissues of wild-type and arsenic (+ 3 oxidation state) methyltransferase-knockout mice

Arsenic methyltransferase(As3mt) catalyzes the conversion of inorganic arsenic(i As) to its methylated metabolites, including toxic methylarsonite(MAs~Ⅲ) and dimethylarsinite(DMAs~Ⅲ). Knockout(KO) of As3 mt was shown to reduce the capacity to methylate i As in mice. However, no data are available on the oxidation states of As species in tissues of these mice. Here, we compare the oxidation states of As species in tissues of male C57BL/6 As3mt-KO and wild-type(WT) mice exposed to arsenite(iA s~Ⅲ) in drinking water. WT mice were exposed to50 mg/L As and As3mt-KO mice that cannot tolerate 50 mg/L As were exposed to 0, 15, 20, 25 or30 mg/L As. iA s~Ⅲaccounted for 53% to 74% of total As in liver, pancreas, adipose, lung, heart, and kidney of As3mt-KO mice; tri- and pentavalent methylated arsenicals did not exceed 10% of total As. Tissues of WT mice retained iA s and methylated arsenicals: iA s~Ⅲ, MAs~Ⅲand DMAs~Ⅲ represented 55%‐68% of the total As in the liver, pancreas, and brain. High levels of methylated species, particularly MAs~Ⅲ, were found in the intestine of WT, but not As3mt-KO mice,suggesting that intestinal bacteria are not a major source of methylated As. Blood of WT mice contained significantly higher levels of As than blood of As3mt-KO mice. This study is the first to determine oxidation states of As species in tissues of As3mt-KO mice. Results will help to design studies using WT and As3mt-KO mice to examine the role of iA s methylation in adverse effects of iA s exposure.     

Alterations of endogenous metabolites in urine of rats exposed to decabromodiphenyl ether using metabonomic approaches

There is large usage of polybrominated diphenyl ethers （PBDEs） especially for decabromodiphenyl ether （BDE-209, Deca-BDE） in controlling the risks of fire. The toxicological effects of PBDEs are worth being concerned about. Female SD rats were daily gavaged with BDE-209 ether at the dose of 100 mg/kg for 20 days. Histological observation was performed for the screening of the target organs for BDE-209 exposure. The distribution and metabolism of PBDEs in the exposed main organs were evidenced by HRGC-HRMS. Alterations of the endogenous metabolite concentrations in urine were investigated using metabonomic approaches based on IH NMR spectrum. Histopathological changes including serious edema in kidney, hepatocellular spotty necrosis and perivasculitis in liver indicated that BDE-209 caused potential influences on endogenous metabolism in the exposed liver and the kidney. BDE-209 was found to be highly accumulated in lipid, ovary, kidney and liver after 20 days＇ exposure. Occurrence of other lower brominated PBDEs in the rats demonstrated that reductive debromination process happened in vivo. Hydroxylated and methoxylated-BDEs, as metabolism products, were also detected in the rat tissues. A total of 12 different endogenous metabolites showed obvious alterations in urine from the exposed rats, indicating the disturbance of the corresponding internal biochemical processes induced by BDE-209 exposure. These findings in vivo suggested the potential health risk might be of concern due to the toxicological effects of BDE-209 as a ubiquitous compound in the environment.     

Studies on metabolism of 2-naphthylamine and its activation mechanism

2-naphthylamine was incubated with induced rat liver microsome S9 preparation and the metabolites were separated through HPLC. The following products were identified: 2-amino-5-naphthol, 2-amino-6-naphthol, 2-amino-7-naphthol and 2-amino-8-naphthol. The yields of these four metabolites are varying in quantity, and the relative contents of 2-amino-8-, -5-, -6- and -7-naphthol are 52.6%, 28.5%, 14.0% and 4.9% respectively. These results are consistent with the quantitative HMO calculation and inference based upon Di-region theory, i.e., the metabolisms of aryl amines on extra-ring (assigned the ring without the substituent of amino group) are through the epoxidation and then NIH shift, but are not the direct hydroxylation in the formation of phenols. It is shown that both the amino group and the carbon atoms on the extra-ring play duality roles of activation and detoxification in metabolism.     

Method to assess component contribution to toxicity of complex mixtures: Assessment of puberty acquisition in rats exposed to disinfection byproducts

A method based on regression modeling was developed to discern the contribution of component chemicals to the toxicity of highly complex, environmentally realistic mixtures of disinfection byproducts(DBPs). Chemical disinfection of drinking water forms DBP mixtures.Because of concerns about possible reproductive and developmental toxicity, a whole mixture(WM) of DBPs produced by chlorination of a water concentrate was administered as drinking water to Sprague–Dawley(S–D) rats in a multigenerational study. Age of puberty acquisition,i.e., preputial separation(PPS) and vaginal opening(VO), was examined in male and female offspring, respectively. When compared to controls, a slight, but statistically significant delay in puberty acquisition was observed in females but not in males. WM-induced differences in the age at puberty acquisition were compared to those reported in S–D rats administered either a defined mixture(DM) of nine regulated DBPs or individual DBPs. Regression models were developed using individual animal data on age at PPS or VO from the DM study. Puberty acquisition data reported in the WM and individual DBP studies were then compared with the DM models. The delay in puberty acquisition observed in the WM-treated female rats could not be distinguished from delays predicted by the DM regression model, suggesting that the nine regulated DBPs in the DM might account for much of the delay observed in the WM. This method is applicable to mixtures of other types of chemicals and other endpoints.     

Coping with the environmental effects of point-source discharges

Current methods for assessing the environmental impact of point source discharges in the United States are reviewed. The emphasis of many assessments of damage from pollution is shifting from chemical or technological standards to biological standards, i.e., the protection of biological integrity in the receiving system. The importance of integrating information from chemical measurements, toxicity tests, and field surveys in impact assessment is re-emphasized. Ways in which environmental professionals can improve methods for assessing damage from pollution and its control are discussed.     

Simultaneous size characterization and mass quantification of the in vivo core-biocorona structure and dissolved species of silver nanoparticles

Size characterization of silver nanoparticles with biomolecule corona(AgNP@BCs) and mass quantification of various silver species in organisms are essential for understanding the in vivo transformation of Ag NPs. Herein, we report a versatile method that allows simultaneous determination of the size of AgNP@BCs and mass concentration of various silver species in rat liver. Both particulate and ionic silver were extracted in their original forms from the organs by alkaline digestion, and analyzed by size exclusion chromatography combined with inductively coupled plasma mass spectrometry(SEC-ICP-MS). While the silver mass concentrations were quantified by ICP-MS with a detection limit of 0.1 μg/g, the effective diameter of AgNP@BCs was determined based on the retention time in SEC separation with size discrimination of 0.6-3.3 nm. More importantly, we found that the BC thickness of AgNP@BCs is core size independent, and a linear correlation was found between the effective diameter and core diameter of AgNP@BCs in extracted tissues, which was used to calibrate the core diameter with standard deviations in the range of 0.2-1.1 nm. The utility of this strategy was demonstrated through application to rat livers in vivo. Our method is powerful for investigating the transformation mechanism of Ag NPs in vivo.     

Alpha-tocopherol supplementation on chromium toxicity: a study on rat liver and kidney cell membrane

Membrane damage is one of the important consequence of chromium,an environmental toxicant,to produce cytotoxicity.α-tocopherol,a membrane protectant can be used to reduce the chromium-induced membrane damage.In the present study,the impact of chromium in presence and absence of α-tocopherol was studied on plasma membrane of live and kidney in male Wistar rats(80-100g body weight).Significant increase in membrane cholesterol level as well as significant decrease in membrane phospholipid level in chromium exposed(0.8mg/100g body weight/d,i.p.,for 4 weeks)animals suggest structural alteration of both liver and kidney plasma memebrane.The alkaline phosphatase,total ATPase and Na^ -K^ -ATPase activities of plasma membrane were significantly decreased in both liver and kidney after chromium treatment.However,α-tocopherol (30mg/100g diet)supplementation can restrict the changes in these membrane-bound enzyme activities.Thus,the usefulness of dietary supplementation of α-tocopherol to restrain the chromium-induced membrane damage is suggested.     

Morphology of single inhalable particle in the air polluted city of Shijiazhuang, China

In the typical air polluted city of Shijiazhuang,single inhalable particle samples in non-heating period,heating pened,dust storm days,and snowy days were collected and detected by SEM/EDS(scanning electron microscopy and energy dispersive X-ray spectrometry).The particle morphology was characterized by the 6 shape clusters,which areirregular square,agglomerate,sphere,floccule,column or stick,and unknown,by quantitative order.The irregular square particles are common in all kinds of samples;sphere particles are more,and column or stick are less in winter samples;in the wet deposit samples,agglomerate and floccule particles are not found.The surface of most particles is coarse with fractal edge,which can provide suitable chemical reaction bed in the polluted atmospheric environment.New formed calcium crystal is found to demonstrate the existence of neutralized reaction,explaining the reason for the high SO2 emission and low acid rain frequency in Shijiazhuang.The three sorts of surface patterns of spheres are smooth,semi-smooth,and coarse,corresponding to the element of Si-dominant,Si-Al-dominant,and Fe-dominant.The soot particle is present as floccule with average size around 10 μm,considerably larger than the former reported results,but wrapped or captured with other fine particles to make its appearance unique and enhance its toxicity potentially.The new formed calcium crystal,the 3 sorts of sphere surface patterns,and the unique soot appearance represent the single inhalable particle's morphology characteristics in Shijiazhuang City.     

Uptake of radioiodide by Paenibacillus sp., Pseudomonas sp., Burkholderia sp. and Rhodococcus sp. isolated from a boreal nutrient-poor bog

Radionuclides, like radioiodine(~(129)I), may escape deep geological nuclear waste repositories and migrate to the surface ecosystems. In surface ecosystems, microorganisms can affect their movement. Iodide uptake of six bacterial strains belonging to the genera Paenibacillus,Pseudomonas, Burkholderia and Rhodococcus isolated from an acidic boreal nutrient-poor bog was tested. The tests were run in four different growth media at three temperatures. All bacterial strains removed iodide from the solution with the highest efficiency shown by one of the Paenibacillus strains with 99% of iodide removed from the solution in one of the used growth media. Pseudomonas, Rhodococcus and one of the two Paenibacillus strains showed highest iodide uptake in 1% yeast extract with maximum values for the distribution coefficient(K_d) ranging from 90 to 270 L/kg DW. The Burkholderia strain showed highest uptake in 1% Tryptone(maximum K_d170 L/kg DW). The Paenibacillus strain V0-1-LW showed exceptionally high uptake in 0.5% peptone + 0.25% yeast extract broth(maximum K_d 1,000,000 L/kg DW). Addition of 0.1% glucose to the 0.5% peptone + 0.25% yeast extract broth reduced iodide uptake at 4℃ and 20℃ and enhanced iodide uptake at 37℃ compared to the uptake without glucose. This indicates that the uptake of glucose and iodide may be competing processes in these bacteria. We estimated that in in situ conditions of the bog,the bacterial uptake of iodide accounts for approximately 0.1%–0.3% of the total sorption of iodide in the surface, subsurface peat, gyttja and clay layers.     

Elevated atmospheric CO2 and drought effects on leaf gas exchange properties of barley

Atmospheric CO₂ concentration (C_a) is rising, predicted to cause global warming, and alter precipitation patterns. During 1994, spring barley (Hordeum vulgare L. cv. Alexis) was grown in a strip-split-plot experimental design to determine the effects that the main plot C_a treatments [A: Ambient at 370 μmol (CO₂) mol⁻¹; E: Enriched with free-air CO₂ enrichment (FACE) at ∼550 μmol (CO₂) mol⁻¹] had on several gas exchange properties of fully expanded sunlit primary leaves. The interacting strip-split-plot irrigation treatments were Dry or Wet [50% (D) or 100% (W) replacement of potential evapotranspiration] at ample nitrogen (261 kg N ha⁻¹) and phosphorous (29 kg P ha⁻¹) fertility. Elevated C_a facilitated drought avoidance by reducing stomatal conductance (g_s) by 34% that conserved water and enabled stomata to remain open for a longer period into a drought. This resulted in a 28% reduction in drought-induced midafternoon depression in net assimilation rate (A). Elevated C_a increased A by 37% under Dry and 23% under Wet. Any reduction in A under Wet conditions occurred because of nonstomatal limitations, whereas under Dry it occurred because of stomatal limitations. Elevated C_a increased the diurnal integral of A (A′) that resulted in an increase in the seasonal-long integral of A′ (A″) for barley leaves by 12% (P = 0.14) under both Dry and Wet - 650, 730, 905 and 1020 ± 65 g (C) m⁻² y⁻¹ for AD, ED, AW and EW treatments, respectively. Elevated C_a increased season-long average dry weight (DW_S; crown, shoots) by 14% (P = 0.02), whereas deficit irrigation reduced DW_S by 7% (P = 0.06), although these values may have been affected by a short but severe pea aphid [Acyrthosiphon pisum (Harris)] infestation. Hence, an elevated-C_a-based improvement in gas exchange properties enhanced growth of a barley crop.     

Chemical composition and properties of ashes from combustion plants using Miscanthus as fuel

Miscanthus giganteus is one of the energy crops considered to show potential for a substantial contribution to sustainable energy production. In the literature there is little data available about the chemical composition of ashes from the combustion of Miscanthus and practically no data about their physical properties. However, for handling, treatment and utilization of the ashes this information is important. In this study ashes from two biomass combustion plants using Miscanthus as fuel were investigated. The density of the ashes was 2230 ± 35 kg/m~3, which was similar to the density of ashes from straw combustion. Also the bulk densities were close to those reported for straw ashes. The flowability of the ashes was a little worse than the flowability of ashes from wood combustion. The measured heavy metal concentrations were below the usual limits for utilization of the ashes as soil conditioner. The concentrations in the bottom ash were similar to those reported for ash from forest residue combustion plants. In comparison with cyclone fly ashes from forest residue combustion the measured heavy metal concentrations in the cyclone fly ash were considerably lower. Cl-, S and Zn were enriched in the cyclone fly ash which is also known for ashes from wood combustion. In comparison with literature data obtained from Miscanthus plant material the concentrations of K, Cl-and S were lower.This can be attributed to the fact that the finest fly ash is not collected by the cyclone de-dusting system of the Miscanthus combustion plants.     

Potential ammonia emission from flag leaves of paddy rice (Oryza sativa L. cv. Koshihikari)

The present study aimed to investigate the potential ammonia (NH₃) emission from flag leaves of paddy rice (Oryza sativa L. cv. Koshihikari). The study was conducted at a paddy field in central Japan that was designed as a free-air CO₂ enrichment (FACE) facility for paddy rice. A dynamic chamber method was used to measure the potential NH₃ emissions. The air concentrations of NH₃ at two heights (2 and 6 m from the ground surface) were measured using a filter-pack method, and the exchange fluxes of NH₃ of the whole paddy field were calculated using a gradient method. The flag leaves showed potential NH₃ emissions of 25-38 ng N cm⁻² h⁻¹ in the daytime from the heading to the maturity stages, and they showed potentials of approximately 22 ng N cm⁻² h⁻¹, even in the nighttime, at the heading and mid-ripening stages. The exchange fluxes of NH₃ of the whole paddy field in the daytime were net emissions of 0.9-3.9 g N ha⁻¹ h⁻¹ whereas the exchange fluxes of NH₃ in the nighttime were approximately zero.     

Effects of a municipal effluent on the freshwater mussel Elliptio complanata following challenge with Vibrio anguillarum

The purpose of this study was to examine the cumulative effects of exposure to a pathogenic bacteria and municipal effluent in the freshwater mussel Elliptio complanata. Mussels were exposed to increasing concentrations of an ozone-treated effluent at 15°C for 7 days. A sub-group of mussels was inoculated with Vibrio anguillarum and exposed to the same conditions as above.After the exposure period, mussels were collected to assess hemocyte count and viability,immunocompetence(phagocytosis and nitrite production), oxidative stress/inflammation(cyclooxygenase and lipid peroxidation) and oxygen radical/xenobiotic scavenging activity(metallothioniens, glutathione S-transferase). The results showed that mussels exposed to municipal effluent had increased hemocyte counts, phagocytosis, nitrites, lipid peroxidation and metallothioneins. In the inoculated mussels, the same responses were observed, in addition to cyclooxygenase and glutathione S-transferase activities. Multivariate analyses revealed that(1)the response pattern changed with effluent concentration, where increased responses observed at low effluent concentrations(10%, V/V) were attenuated at higher effluent concentrations,(2)the effluent produced more pronounced changes in lipid peroxidation, metallothionein and hemocyte viability, and(3) the simultaneous presence of V. anguillarum led to more important changes in hemocyte count and viability and nitrite levels. In conclusion, the presence of V.anguillarum could alter the response of mussels to municipal effluent, which could lead to increased inflammation in mussels.     

Enhanced photoelectrocatalytic degradation of ammonia by in situ photoelectrogenerated active chlorine on TiO2 nanotube electrodes

TiO_2 nanotube(Ti NT) electrodes anodized in fluorinated organic solutions were successfully prepared on Ti sheets. Field-emission scanning electron microscopy(FE-SEM) and X-ray diffraction(XRD) were performed to characterize the TiNT electrodes. The linear voltammetry results under irradiation showed that the TiNT electrode annealed at 450°C presented the highest photoelectrochemical activity. By combining photocatalytic with electrochemical process, a significantly synergetic effect on ammonia degradation was observed with Na_2SO_4 as supporting electrolyte at pH 10.7. Furthermore, the photoelectrocatalytic efficiency on the ammonia degradation was greatly enhanced in presence of chloride ions without the limitation of pH. The degradation rate was improved by 14.8 times reaching 4.98 × 10~(-2) min~(-1) at pH 10.7 and a faster degradation rate of 6.34 × 10~(-2) min~(-1)was obtained at pH 3.01. The in situ photoelectrocatalytic generated active chlorine was proposed to be responsible for the improved efficiency. On the other hand, an enhanced degradation of ammonia using TiNT electrode fabricated in fluorinated organic solution was also confirmed compared to TiNT electrode anodized in fluorinated water solution and TiO_2 film electrode fabricated by sol–gel method. Finally, the effect of chloride concentration was also discussed.     

Differences in nitrite-oxidizing communities and kinetics in a brackish environment after enrichment at low and high nitrite concentrations

Nitrite accumulation in shrimp ponds can pose serious adverse effects to shrimp production and the environment.This study aims to develop an effective process for the enrichment of ready-to-use nitrite-oxidizing bacteria(NOB)inocula that would be appropriate for nitrite removal in brackish shrimp ponds.To achieve this objective,the effects of nitrite concentrations on NOB communities and nitrite oxidation kinetics in a brackish environment were investigated.Moving-bed biofilm sequencing batch reactors and continuous moving-bed biofilm reactors were used for the enrichment of NOB at various nitrite concentrations,using sediment from brackish shrimp ponds as seed inoculum.The results from NOB population analysis with quantitative polymerase chain reaction(q PCR)show that only Nitrospira were detected in the sediment from the shrimp ponds.After the enrichment,both Nitrospira and Nitrobacter coexisted in the reactors controlling effluent nitrite at 0.1 and 0.5 mg-NO_2~--N/L.On the other hand,in the reactors controlling effluent nitrite at 3,20,and 100 mg-NO_2~--N/L,Nitrobacter outcompeted Nitrospira in many orders of magnitude.The half saturation coefficients(Ks)for nitrite oxidation of the enrichments at low nitrite concentrations(0.1 and 0.5 mg-NO_2~--N/L)were in the range of 0.71–0.98 mg-NO_2~--N/L.In contrast,the Ksvalues of NOB enriched at high nitrite concentrations(3,20,and 100 mg-NO_2~--N/L)were much higher(8.36–12.20 mg-NO_2~--N/L).The results suggest that the selection of nitrite concentrations for the enrichment of NOB inocula can significantly influence NOB populations and kinetics,which could affect the effectiveness of their applications in brackish shrimp ponds.     

Biodegradation of pendimethalin by Bacillus subtilis Y3

A bacterium strain Y3,capable of efficiently degrading pendimethalin,was isolated from activated sludge and identified as Bacillus subtilis according to its phenotypic features and 16 S rRNA phylogenetic analysis.This strain could grow on pendimethalin as a sole carbon source and degrade 99.5%of 100 mg/L pendimethalin within 2.5 days in batch liquid culture,demonstrating a greater efficiency than any other reported strains.Three metabolic products,6-aminopendimethalin,5-amino-2-methyl-3-nitroso-4-(pentan-3-ylamino) benzoic acid,and 8-amino-2-ethyl-5-(hydroxymethyl)-1,2-dihydroquinoxaline-6-carboxylic acid,were identified by HPLC-MS/MS,and a new microbial degradation pathway was proposed.A nitroreductase catalyzing nitroreduction of pendimethalin to 6-aminopendimethalin was detected in the cell lysate of strain Y3.The cofactor was nicotinamide adenine dinucleotide phosphate(NADPH) or more preferably nicotinamide adenine dinucleotide(NADH).The optimal temperature and pH for the nitroreductase were 30℃ and 7.5,respectively.Hg~(2+),Ni~(2+),Pb~(2+),Co~(2+),Mn~(2+) Cu~(2+),Ag~+,and EDTA severely inhibited the nitroreductase activity,whereas Fe~(2+),Mg~(2+),and Ca~(2+) enhanced it.This study provides an efficient pendimethalin-degrading microorganism and broadens the knowledge of the microbial degradation pathway of pendimethalin.     

Comparative cytotoxicity of nanoparticles and ions to Escherichia coli in binary mixtures

The objective of this study was to understand toxicity of mixture of nanoparticles (NPs) (ZnO and TiO₂) and their ions to Escherichia coli. Results indicated the decrease in percentage growth of E. coli with the increase in concentration of NPs both in single and mixture setups. Even a small concentration of 1 mg/L was observed to be significantly toxic to E. coli in binary mixture setup (exposure concentration: 1 mg/L ZnO and 1 mg/L TiO₂; 21.15% decrease in plate count concentration with respect to control). Exposure of E. coli to mixture of NPs at 1000 mg/L (i.e., 1000 mg/L ZnO and 1000 mg/L TiO₂) resulted in 99.63% decrease in plate count concentration with respect to control. Toxic effects of ions to E. coli were found to be lesser than their corresponding NPs. The percentage growth reduction was found to be 36% for binary mixture of zinc and titanium ions at the highest concentration (i.e., 803.0 mg/L Zn and 593.3 mg/L Ti where ion concentrations are equal to the Zn ions present in 1000 mg/L ZnO NP solution and Ti^+ 4 ions present in 1000 mg/L TiO₂ NP solution). Nature of mixture toxicity of the two NPs to E. coli was found to be antagonistic. The alkaline phosphatase (Alp) assay indicated that the maximum damage was observed when E. coli was exposed to 1000 mg/L of mixture of NPs. This study tries to fill the knowledge gap on information of toxicity of mixture of NPs to bacteria which has not been reported earlier.     

Biodegradation of indole by a newly isolated Cupriavidus sp. SHE

Indole, a typical nitrogen heterocyclic aromatic pollutant, is extensively spread in industrial wastewater. Microbial degradation has been proven to be a feasible approach to remove indole, whereas the microbial resources are fairly limited. A bacterial strain designated as SHE was isolated and found to be an efficient indole degrader. It was identified as Cupriavidus sp. according to 16S rRNA gene analysis. Strain SHE could utilize indole as the sole carbon source and almost completely degrade 100 mg/L of indole within 24 hr. It still harbored relatively high indole degradation capacity within pH 4–9 and temperature 25°C–35°C. Experiments also showed that some heavy metals such as Mn^2 +, Pb^2 + and Co^2 + did not pose severe inhibition on indole degradation. Based on high performance liquid chromatography–mass spectrum analysis, isatin was identified as a minor intermediate during the process of indole biodegradation. A major yellow product with m/z 265.0605 (C₁₅H₈N₂O₃) was generated and accumulated, suggesting a novel indole conversion pathway existed. Genome analysis of strain SHE indicated that there existed a rich set of oxidoreductases, which might be the key reason for the efficient degradation of indole. The robust degradation ability of strain SHE makes it a promising candidate for the treatment of indole containing wastewater.