Uptake of Tributyltin into Willow Trees

BACKGROUND: Organotins have been used world-wide as antifoulants in ship paints. Repeatedly, severe effects on aquatic species have resulted. The use of organotins for this purpose was ruled out, and dumping of contaminated harbor sludge into the sea was prohibited. Land-based dumping is seen as an alternative. OBJECTIVE: This study investigates sorption, uptake and translocation of tributyltin (TBT) to willow trees in order to evaluate phytoremediation as treatment option. The study considers the influence of pH on the plant uptake of organotins. EXPERIMENTAL SET-UP: Chemicals investigated were the weak base tributyltin chloride (TBTCl) and the neutral tributyltin hydride (TBTH). Organotins were extracted from solution and plant material with toluene, and analyzed as tin by AAS with graphite oven. The pH in solution varied from pH 4 to pH 7. The sorption to living and dead roots, stems and leaves was measured in shaking experiments. The uptake into intact trees was measured at nominal levels of 1 and 10 mg TBT/l for TBTH and TBTCl at low and high pH. RESULTS: The sorption to roots and leaves dropped for dead tissue, but did not vary much with pH. The sorption to stems increased for dead stems and with pH. The solubility of TBTCl in water was below 10 mg/l and lowest at pH 4. Concentrations of TBTCl and TBTH in solutions with trees dropped rapidly to low values. Highest TBT contents in trees were found in roots and lower stems. The concentrations followed the concentrations in solution. The pH had only a small effect on the plant uptake of TBTCl, and no effect on the uptake of TBTH. No effective translocation to higher stems or leaves was found. DISCUSSION: An ion trap mechanism that accumulates the weak base TBTCl in the xylem sap of plants and leads to upward translocation could not be detected. Neither TBTCl at low or high pH, nor the neutral lipophilic chemical TBTH, were translocated effectively to leaves. The TBT+ cation sorbed strongly to plant tissue. The exact mechanism for the strong sorption of the cation is unknown, but similar effects have been observed for algae, liposomes and isolated biomembranes. CONCLUSIONS: Both the uptake of the neutral TBTH and the uptake of the neutral molecule form of TBTCl into willows was as is to be expected from theory. The cation TBT+ showed an unexpected behavior which has been observed before. No ion trap occurs, and the phytoextraction of TBT is not feasible. OUTLOOK: Planting trees, or other appropriate vegetation, could have a beneficial remediation effect by aeration of the TBT-contaminated soil or sludge. In a follow-up paper, the toxicity of TBT to willow trees will be described.     

海洋环境中的有机锡及其对海洋生物的影响

本文综述了有机锡化占物对海洋环境的冲击及其对海洋生物的影响,介绍了有机锡化合物的分析方法,讨论了有机锡污染的预防和控制措施,并展望了有机锡研究的前景。     

Growth of Plants on TBT-contaminated Harbour Sludge and Effect on TBT Removal (10 pp)

Background Worldwide, large amounts of sediments have to be dredged annually from waterways and harbours. These sediments are sometimes polluted with a variety of toxic compounds. In some countries, including Belgium, the load with the biocide tributyltin (TBT) from ship coatings prohibits the dumping of harbour sludge into the sea. Land-based dumping is a commonly used alternative. Objective This research investigated the feasibility to use land-deposited harbour sludge for plant production. In a field trial, the growth of 38 more or less salt-tolerant plant species on low and high TBT-contaminated sediments was studied. The elimination of TBT from sludge with and without vegetation was compared. The uptake of TBT and its degradation products di- and monobutyltin (DBT and MBT) into harvest products under field conditions was determined. - Experimental Set-up. Sediments dredged in May 2003 from the brackish waters of the port of Antwerp were analysed in the laboratory for soil texture, pH, electroconductivity, sodium, magnesium, potassium, calcium, ammonium, nitrate, total nitrogen, chloride, sulphur and the organotins TBT, DBT and MBT. The sediments were lagooned for one year to dewater, desalinate and improve their structure. Salt-tolerant domestic and wild plants were selected and sown in May 2004. In August 2004, plants were harvested and the produced biomass was determined. Samples were taken from vegetated and non-vegetated top and bottom sediments and from plants growing above soil and analysed for TBT, DBT and MBT. Results The fresh sediments showed a good supply with nutrients and a neutral pH, but were rather saline (EC 14 mS cm-1 of the saturated paste extract). The salinity decreased to 3.7 mS cm-1 during lagoonation. The high and the low contaminated sediment had initially 43 and 1.6 mg TBT kg-1 dry weight, respectively. Besides TBT, several other contaminants were present in the sediments at critical levels. The biomass production of the plant species from the field trial ranged from 0.2 to 13 tons dry mass per hectare. Plants performing excellently were barley, sorghum, rape seed, a clover/grass mix and reed. If at all, a positive influence of TBT on plant growth was seen. TBT was degraded significantly faster (>40%) below barley. The uptake of TBT, DBT and MBT into stem and leaves of reed, grass and clover was very low, but measurable and not related to concentrations in soil. No uptake of TBT or its metabolites into corn of barley was found. Discussion This study confirmed former results: the toxicity of TBT to higher plants is low, and even high levels in soils would not be a hindrance for crop production. The removal of TBT seemed to be increased by both lagooning and plant growth, although the target values for sea dumping in use in certain European countries were not reached. A plausible explanation for the faster degradation of TBT under vegetation is that oxygen is a limiting factor, and plants dewater the soil, thus aerating it. The uptake of the organotins TBT, DBT and MBT into harvest products is probably due to attached soil particles. Conclusions To summarize, barley was the optimal species: it grew very well despite the salinity of the dredged sediments, it had a significantly positive effect on TBT removal; it showed no measurable uptake of TBT or the other butyltins into the harvested product; and it is a cash crop well established in European agriculture. Outlook The amounts of dredged sediments are high, and good soils are becoming increasingly rare. The feasibility of using dredged sediments for non-food production, such as energy crops, should be investigated by a critical risk assessment.     

Assessment of Antifouling Effectiveness of two Natural Product Antifoulants by Attachment Study with Freshwater Bacteria (7 pp)

- Goal, Scope, Background. The traditional solution for keeping unwanted organisms from attaching to submerged surfaces is to apply anti-fouling coatings. The most common antifoulant was tributyltin (TBT). TBT systems were highly effective but were also toxic to non-target organisms. The use of the TBT based coatings will be completely banned by January 1, 2008. Therefore, there is an urgent need to seek out suitable non-toxic alternatives. Methods The aim of this work was to evaluate the effectiveness of capsaicin and zosteric acid as natural product antifoulants (NPAs) in deterring bacterial attachment. Two fresh water bacteria systems Pseudomonas putida (Pp) and bacteria isolated from Lake Erie (LE) were used to assess the attachment when the NPAs dispersed in the water. Effectiveness was ascertained based on the decrease in microbial attachment, limited toxicity, and minimum alteration of the coatings properties. Results and Discussion A significant inhibition of bacteria attachment was achieved when aqueous capsaicin concentration was increased from 0 to 40 mg/L. For instance, after 14 days the LE system depicted 93.5% and 98.5% less biofilm coverage for 20 mg/L and 40 mg/L capsaicin, respectively when compared to systems without NPA. Biofilm coverage was reduced by 92.5% and 98.2%, respectively with 50 mg/L and 500 mg/L zosteric acid. Conclusions Both capsaicin and zosteric acid was effective at preventing bacteria attachment. As the NPA aqueous concentration increased, biofilm formation decreased. Evaluating changes in aqueous pH, conductivity, dissolved oxygen, aqueous microbial population and biofilm formation suggested that the primary antifoulant mechanism of these two NPAs was to block the bacteria's active sites versus posing a lethal level. Recommendation and Perspective From the attachment study, zosteric acid appeared to be more effective in preventing bacterial attachment when the NPAs were dispersed in the aqueous environment. For practical applications, the antifoulant needs to be incorporated into a coating and have a slow release rate. Thus the ability to successfully incorporate zosteric acid into a coating, without deterring bacterial attachment, needs to be investigated.     

Phytotoxicity of Cyanide to Weeping Willow Trees

Background Cyanide is found predominantly in industrial effluents generated by metallurgical operations. It is an extremely toxic compound, so that problems and catastrophic accidents have recently occurred all around the globe. The goal of this study was to determine the toxicity of cyanide to a Chinese willow species, and to determine the removal capacity. Methods The toxicity of potassium cyanide (KCN) to weeping willow trees (Salix babylonica L.) was tested. The normalized, relative transpiration of the plants was used to determine the phytotoxicity of cyanide. The cyanide removal capacity of weeping willows was also determined. Results and Discussion In hydroponic solution, no chlorosis of leaves and only a small reduction in normalized relative transpiration was observed when weeping willows were exposed to low doses of cyanide (0.93 mg CN/L). Severe signs of toxicity were found for the treatment groups exposed to higher doses of cyanide (9.3 mg CN/L). Weeping willows grown in sandy soils survived the entire period (216 hours) without any toxic effect when irrigated with low doses of cyanide (3.72 mg CN/L). High doses of cyanide (18.6 mg CN/L) in irrigation water were fatal for the weeping willows within 216 hours. EC50 values for a 50% inhibition of the transpiration of the trees were estimated to be between 3.27 and 8.23 mg CN/L, depending on the duration of the exposure. Conclusions The results obtained for the Chinese willow species Salix babylonica were very similar to those obtained for the European species S. viminalis in earlier studies. Phytotoxic effects were only found at high doses of cyanide. A large proportion of applied cyanide was removed from the contaminated media in the presence of weeping willows. This gives rise to the conclusion that the metabolism of cyanide by weeping willows is possible. Recommendations and Outlook Cyanide elimination with trees seems to be a feasible option for cleaning soils and water contaminated with cyanide. A full-scale treatment has been installed in Denmark. For phytoremediation projects in China, weeping willow could be a suitable species. The tree can tolerate and remove cyanide, and it is a native Chinese species. Besides, the tree is of outstanding beauty and is planted as a common park tree in many parts of the world.     

Metabolic responses of weeping willows to selenate and selenite

Goal, Scope and Background Selenium (Se) is one of the most widely distributed elements of the earth’s crust at low concentrations. The extensive use of Se-containing chemicals due to anthropogenic activities has increased the ecological risk to environmental compartments. Plants, under unfavorable environmental conditions, often increase the formation of reactive oxygen species (ROS), and consequently plant antioxidant enzymatic systems have been proposed to be important in plant stress tolerance. The goal of this study was to find out the metabolic responses of plants to Se, to provide quantitative information whether exogenous Se has a beneficial role in plants, and to investigate the potential of vegetation management of Se for potential phytoremediation. Material and Methods Pre-rooted plants of weeping willows (Salix babylonica L.) were grown hydroponically in growth chambers and treated with Na₂SeO₄ or Na₂SeO₃ at 24.0 ± 1°C for 168 h. Five different treatment concentrations were used, ranging from 0.44 to 8.72 mg Se/L for the treatments exposed to SeO₄²⁻ and from 0.50 to 10.0 mg Se/L for the treatments exposed to SeO₃²⁻, respectively. Transpiration rates, soluble protein contents and antioxidative enzyme activities of the plants were monitored to evaluate toxicity from exogenous Se exposure. At the end of the study, total Se in the hydroponic solution was analyzed by hydride generation-atomic fluorescence spectrometry (HG-AFS). Results Both chemical forms of Se at low concentrations showed growth-promoting effects on plants. A significant decrease of transpiration rates and of soluble protein contents of plants was observed at higher Se concentrations after 168 h of exposure. Measurable change of superoxide dismutases (SOD) activity in leaves was only detected under high Se treatments. Catalase (CAT) activity was significantly affected by the Se application. Slight change of peroxidase (POD) activity was measured in all treatments, whereas significant inhibition of POD activity was detected for the plants exposed to SeO₃²⁻ of 10.0 mg Se/L. Se-induced stress appeared in all treatments, thus resulting in measurable increase of glutathione peroxidase (GSH-Px) activity of the plants. Although both chemical forms of Se were taken up by weeping willows efficiently, their uptake rates were different. Discussion Of all measured parameters, POD and CAT activities in leaves were noted the most sensitive indicator for the plants exposed to SeO₄²⁻ and SeO₃²⁻, respectively. Deleterious effects on plant physiological functions due to Se application were not observed over 168 h of exposure. This is largely due to the fact that well-established antioxidant enzymatic systems in plants and higher activities of GSH-Px largely reduced the negative effects on plants; SeO₃²⁻ caused much more severe stress to plants than SeO₄²⁻ at higher Se application rates. The uptake mechanisms between the two chemical species were quite different. Conclusions Neither visible toxic symptoms nor metabolic lesions were observed at low concentrations of Se, probably due to the effective established enzymatic systems in weeping willows. All selected parameters for toxicity determination were significantly correlated to Se application, but metabolic responses of plants to SeO₄²⁻ and SeO₃²⁻ were quite different. GSH-Px in leaves was probably the principle enzyme responsible for stress reduction from Se exposure. Due to their different chemical properties, weeping willows showed a faster uptake rate for SeO₄²⁻ than for SeO₃²⁻. Recommendations Exogenous Se has a beneficial role in plants and vegetation management of Se is a potential remediation strategy in cleaning up Se-contaminated sites. Further investigation on the biochemical mechanism of Se metabolism will provide insight to the specific interactions between Se and plants on the molecular level. Perspectives Weeping willow has a sound potential for phytoremediation of Se-contaminated sediment and groundwater because the tree is not only tolerant to Se but also uptakes chemical species from the environment.     

Persistence of methyl <Emphasis Type="Italic">tertiary</Emphasis> butyl ether (MTBE) against metabolism by Danish vegetation

BACKGROUND: Methyl tertiary butyl ether (MTBE) is the second most highly produced industrial chemical in the US and a frequent groundwater pollutant. At the same time, MTBE is quite persistent to biotic and abiotic decomposition. The goal of this study was to find plant species that could degrade MTBE and might be used in phytoremediation. METHODS: Excised roots and leaves (0.3 g) from more than 24 Danish plant species out of 15 families were kept in glass vessels with 25 ml spiked aqueous solution for 2 to 4 days. MTBE concentrations were 1 to 5 mg/L. Samples were taken directly from the solution with a needle and injected to a purge and trap unit. MTBE and the main metabolite, TBA, were measured by GC/FID. RESULTS AND DISCUSSION: Solutions with roots of poplar (Populus robusta) and a willow hybrid (Salix viminalis x schwerinii) produced TBA in trace amounts, probably stemming from bacteria. Significant MTBE reduction (> 10%) was not observed in any of the tests. Leaves from none of the species (trees, grasses and herbs) reduced the concentration of MTBE in the solution and no TBA, nor any other known metabolite of MTBE, was detected. CONCLUSION: It was not possible to find plants capable of efficiently degrading MTBE. This gives rise to the conclusion that plants probably cannot degrade MTBE at all, or only very slowly. RECOMMENDATIONS AND OUTLOOK: For phytoremediation projects, this has, as consequence, that the volatilization by plants (except with genetically engineered plants) is the only relevant removal process for MTBE. For risk assessment of MTBE, degradation by the plant empire is not a relevant sink process.     

Mixture Toxicity of the Antifouling Compound Irgarol to the Marine Phytoplankton Species Dunaliella tertiolecta

This study examined the toxicity of irgarol, individually and in binary mixtures with three other pesticides (the fungicide chlorothalonil, and the herbicides atrazine and 2,4-D), to the marine phytoplankton species Dunaliella tertiolecta. Standard 96-h static algal bioassays were used to determine pesticide effects on population growth rate. Irgarol significantly inhibited D. tertiolecta growth rate at concentrations ≥ 0.27 μ g/L. Irgarol was significantly more toxic to D. tertiolecta than the other pesticides tested (irgarol 96 h EC₅₀ = 0.7 μ g/L; chlorothalonil 96 h EC₅₀ = 64 μ g/L; atrazine 96 h EC₅₀ = 69 μ g/L; 2,4-D 96 h EC₅₀ = 45,000 μ g/L). Irgarol in mixture with chlorothalonil exhibited synergistic toxicity to D. tertiolecta, with the mixture being approximately 1.5 times more toxic than the individual compounds. Irgarol and atrazine, both triazine herbicides, were additive in mixture. The toxicity threshold of 2,4-D was much greater than typical environmental levels and would not be expected to influence irgarol toxicity. Based on these interactions, overlap of certain pesticide applications in the coastal zone may increase the toxicological risk to resident phytoplankton populations.     

Toxicity of carbaryl,diquat dibromide,and fluoranthene,individually and in mixture,to larval grass shrimp,Palaemonetes pugio

This study examined the toxicity of two pesticides (carbaryl and diquat dibromide) and one polycyclic aromatic hydrocarbon (fluoranthene), both singly and in mixture, to grass shrimp larvae (Palaemonetes pugio). These three chemicals are all present in coastal environments and can easily enter estuarine ecosystems. Fluoranthene was the most toxic chemical with a 96-h LC₅₀ value of 32.45 μ g/L, followed by carbaryl (43.02 μ g/L) and diquat dibromide (1624 μ g/L). In the chemical mixture tests, the binary carbaryl/diquat dibromide mixture and the ternary carbaryl/diquat dibromide/fluoranthene mixture had additive results.     

Predicting molybdenum toxicity to higher plants: Influence of soil properties

The effect of soil properties on the toxicity of molybdenum (Mo) to four plant species was investigated. Soil organic carbon or ammonium-oxalate extractable Fe oxides were found to be the best predictors of the 50% effective dose (ED₅₀) of Mo in different soils, explaining > 65% of the variance in ED₅₀ for four species except for ryegrass (26-38%). Molybdenum concentrations in soil solution and consequently plant uptake were increased when soil pH was artificially raised because sorption of Mo to amorphous oxides is greatly reduced at high pH. The addition of sulphate significantly decreased Mo uptake by oilseed rape. For risk assessment, we suggest that Mo toxicity values for plants should be normalised using soil amorphous iron oxide concentrations.     

The critical levels and the maximum metal uptake for wheat and rice plants when applying metal oxides to soil

Abstract

Wheat is more sensitivity to CdO and ZnO compared with rice plant. The yield of wheat decreased by 30% in the presence of 30 ppm Cd, but that of rice plants by only 8%. The critical levels of meal uptake by wheat and rice plants for applying metal oxides to soil (CdO, ZnO, PbO) were determined. The highest concentration obtained for wheat grain was 141 μg/g Cd at the Cd 10,000 ppm in soil. This value is higher the value of 4.97 μg/g for unpolished rice and higher than any other we have seen in the reports for treatment with CdO. Also, as concentration of more than 1.0 μg/g Cd in wheat was obsertced at 5 ppm Cd, while similar concentrations for rice plants were observed at 30 ppm Cd in soil.     

Influence of Phenanthrene on Cadmium Toxicity to Soil Enzymes and Microbial Growth (5 pp)

Background Many contaminated sites contain a variety of toxicants. Risk assessment and the development of soil quality criteria therefore require information on the interaction among toxicants. Interactions between heavy metals are relatively well studied, but little is known about those between heavy metals and polycyclic aromatic hydrocarbons (PAHs). Methods 0.1 mg/kg dry soil phenanthrene alone or phenanthrene plus 10 mg/kg cadmium (Cd) were added to soil to determine the influence of phenanthrene on Cd toxicity to soil enzymes (invertase, urease, dehydrogenase and phosphatase) and microorganisms (fungi, bacteria and actinomycete) in paddy soil. Results and Discussion 0.1 mg/kg phenanthrene did not reduce the number of microorganisms. However, the addition of phenanthrene to soil with Cd enhanced the Cd toxicity to soil enzymes and microorganisms. This deleterious effect was seen to mainly affect the growth of fungi and the activity of invertase, urease and dehydrogenase. The order of combined inhibition of Cd and phenanthrene was fungi>bacteria>actinomycete. Conclusion The presence of phenanthrene might enhance the toxicity of Cd to soil microorganisms. Phenanthrene can easily be used by the soil actinomycetes as a source of carbon and energy and the finding may be supportive to the development of bioremediation techniques.     

Toxicity of synthetic pyrethroid insecticides to the grass shrimp,Palaemonetes pugio,parasitized with the bopyrid isopod,Probopyrus pandalicola

The grass shrimp, Palaemonetes pugio, plays a large role in the marine ecosystem, serving as a vital link in the food web between many other species. Marine parasites such as the bopyrid isopod, Probopyrus pandalicola, reduce shrimp growth and reproductive output and may also cause P. pugio to be more vulnerable to the lethal effects of contaminants. The purpose of this study was to determine the toxicity of resmethrin and bifenthrin on the grass shrimp, P. pugio, infected with the bopyrid isopod, Probopyrus pandalicola. A 96-h static renewal test was conducted to determine the toxicity of the pyrethroid insecticides resmethrin and bifenthrin to grass shrimp, Palaemonetes pugio, parasitized with the bopyrid isopod, Probopyrus pandalicola. The results were then compared to similar tests utilizing unparasitized P. pugio. Parasitized P. pugio had lower 24-h LC₅₀ (1.08 μg/L) and 96-h LC₅₀ (0.43 μg/L) values for resmethrin than unparasitized P. pugio. However, LC₅₀ ratio tests found that there was no significant difference between parasitized and unparasitized shrimp when affected by resmethrin (p = 0.1751 and 0.1108, respectively). In contrast, an LC₁₀ ratio test indicated that there was a significant difference between parasitized and unparasitized P. pugio after 96 h (p < 0.0001). When subjected to bifenthrin, parasitized P. pugio had a higher 24-h LC₅₀ (0.049 μg/L6) than unparasitized P. pugio. The LC₅₀ ratio test established that the effects of bifenthrin on parasitized P. pugio when compared to unparasitized P. pugio were significantly different at 24 h (p = 0.0065). However, there were no significant differences between parasitized and unparasitized after 96 h (p = 0.4229). In conclusion, both resmethrin and bifenthrin are toxic to the grass shrimp, P. pugio, regardless of parasite presence, and parasitized shrimp may be more susceptible to lower doses of resmethrin (when exposed in the field).     

Application of encapsulation (pH-sensitive polymer and phosphate buffer macrocapsules): a novel approach to remediation of acidic ground water

Macrocapsules, composed of a pH-sensitive polymer and phosphate buffer, offer a novel remediation alternative for acidic ground waters. To test their potential effectiveness, laboratory experiments were carried out followed by a field trial within a coal pile runoff (CPR) acidic contaminant plume. Results of traditional limestone and macrocapsule treatments were compared in both laboratory and field experiments. Macrocapsules were more effective than limestone as a passive treatment for raising pH in well water from 2.5 to 6 in both laboratory and field experiments. The limestone treatments had limited impact on pH, only increasing pH as high as 3.3, and armoring by iron was evident in the field trial. Aluminum, iron and sulfate concentrations remained relatively constant throughout the experiments, but phosphate increased (0.15-32 mg/L), indicating macrocapsule release. This research confirmed that macrocapsules may be an effective alternative to limestone to treat highly acidic ground water.     

Factors influencing concentrations of dissolved gaseous mercury (DGM) and total mercury (TM) in an artificial reservoir

The effects of various factors including turbidity, pH, DOC, temperature, and solar radiation on the concentrations of total mercury (TM) and dissolved gaseous mercury (DGM) were investigated in an artificial reservoir in Korea. Episodic total mercury accumulation events occurred during the rainy season as turbidity increased, indicating that the TM concentration was not controlled by direct atmospheric deposition. The DGM concentration in surface water ranged from 3.6 to 160 pg/L, having a maximum in summer and minimum in winter. While in most previous studies DGM was controlled primarily by a photo-reduction process, DGM concentrations tracked the amount of solar radiation only in winter when the water temperature was fairly low in this study. During the other seasons microbial transformation seemed to play an important role in reducing Hg(II) to Hg(0). DGM increased as dissolved organic carbon (DOC) concentration increased (p-value < 0.01) while it increased with a decrease of pH (p-value < 0.01).     

A comparative study on the uptake of polycyclic aromatic hydrocarbons by Anodonta californiensis

Uptake of polycyclic aromatic hydrocarbons (PAHs) by the freshwater bivalve mollusc Anodonta californiensis was examined in the presence and absence of surfactant in order to gain further insight into mixture toxicity and to predict whether certain mixtures have negative and/or positive effects on aquatic organisms. In the presence of surfactant, the uptake of anthracene or chrysene was higher than that of naphthalene, given the same concentration in the solution. In the absence of surfactant, the trend was similar, but the uptakes were increased by approximately 100% compared to those in the presence of surfactant. On the uptake of naphthalene, the presence of anthracene showed only minor influence. The uptake of anthracene was affected by both naphthalene and chrysene. The uptake of chrysene was influenced by neither naphthalene nor anthracene. There was no observable displacement of divalent cations from the surface of the gill membrane by any of the PAHs studied.     

Assessing the toxicity and teratogenicity of pond water in north-central minnesota to amphibians

BACKGROUND: Incidence of amphibian deformities have increased in recent years, especially in the northern region of the United States. While many factors have been proposed as being responsible for generating deformities (e.g., contaminants, ultraviolet radiation [UV], parasites), no single cause has been definitively established. METHODS: To determine whether waterborne chemicals are responsible for amphibian deformities in ponds in north-central Minnesota, we deployed semipermeable membrane devices (SPMDs) in an impacted and a reference site to accumulate lipophilic contaminants. We then exposed native tadpoles (northern leopard frogs; Rana pipiens) to the SPMD extracts combined with two agricultural pesticides (atrazine, carbaryl) at two levels of UV radiation. RESULTS AND DISCUSSION: UV radiation alone caused a slight increase in hatching success and tadpole growth rate. Deformity rate among hatchlings was high following exposure to SPMD extracts from the reference site in the absence of UV, suggesting that chemicals present at this site are broken down by UV to less harmful forms, or become less bioavailable. Conversely, impacted site SPMD extracts caused hatchling deformities only in the presence of UV, suggesting that UV potentiates the teratogenicity of the compounds present there. Impacted site SPMD extracts significantly increased the number of bony triangles among metamorphs, a common deformity observed at this site. The incidence of skin webbings increased significantly with SPMD extracts from both sites as well as with our pesticide control containing atrazine and carbaryl alone. CONCLUSIONS: Higher deformity rates among tadpoles reared in the presence of UV radiation and SPMD extracts from sites where deformities are common indicates a chemical compound (or compounds) in the water at this site may be causing the deformities. RECOMMENDATIONS AND OUTLOOK: It is important to examine the effects of chemical stressors in the presence of other natural stressors (e.g., UV radiation) to gain a better understanding of how multiple stressors work to impact amphibians and amphibian populations.     

Multivariate Statistics of the Pyrolysis Products of High Molecular Components in Pulping Wastewaters to Explain Their Toxicity (6 pp)

Background, Aim and Scope At present, large-scale paper manufacture involves delignification and bleaching by elemental chlorine free (ECF) or totally chlorine free (TCF) processes. The wastewater is purified by secondary treatment (mechanical, chemical and biological) which removes most of the toxic substances from the discharge. However, we found residual toxicity in the high molecular (> 1000 D) matter (HMWM) of the discharge by test of the RET (reverse electronic transfer) inhibition. This fraction consists mainly of polydisperse lignin (LIG) and carbohydrate (CH) macromolecules. Structural units in these molecules are studied by pyrolysis gas chromatography / mass spectrometry (Pyr-GC/MS). In the present work, our aim was to find out those structural units which could explain the RET toxicity of LIG or CH molecules. We compared statistically RET toxicity values of the HMWM samples from treated wastewaters of pilot pulping experiments and intensity variation of the pyrolysis product gas chromatograms of these samples. This application is a novel study procedure. Methods Pyrolysis products (Py-GC/MS results) and inhibition of RET (reverse electronic transport toxicity) as TU50 and TU20 of HMWM (High Molecular Weight Material; Mw > 1000 D) were compared by multivariate statistics. The samples were from laboratory pilot stages of TCF (Totally Chlorine Free) and ECF (Elemental Chlorine Free) manufacture of softwood pulp. Py-GC/MS was done without and with addition of TMAH (Tetra Methyl Ammonium Hydroxide). The name and structure of each abundant fragment compound was identified from its retention time and mass spectrum compared to authentic reference compounds or literature. Four sets of Toxicity Units (TUs) and GC peak areas of the pyrolysis fragments were obtained. The data were normalized by division with LIG (lignin content of each sample). TU values were dependent and the fragment values independent (explanatory) variables in statistical treatments by SPSS system. Separate analyses of correlations, principal components (PCA) and stepwise multiple linear regression (SMLR) were performed from the four sample sets TCF and ECF with and without TMAH. Results and Discussion From the CH fragments, 2-furfural in TCF, and from the LIG fragments, styrene in ECF showed the highest probabilities to originate from source structures of toxicity. Other possible compounds in concern were indicated to be CH fragment 2-methyl-2-cyclopenten-1-one in ECF and LIG fragments 2-methoxy-4-methylphenol, 4,5-dimethoxy-2-methylphenol and 2-methylphenol in TCF.     

Phytoremediation potentials of selected tropical plants for Ethidium bromide

Background, Aims and Scope Research and development has its own benefits and inconveniences. One of the inconveniences is the generation of enormous quantity of diverse toxic and hazardous wastes and its eventual contamination to soil and groundwater resources. Ethidium bromide (EtBr) is one of the commonly used substances in molecular biology experiments. It is highly mutagenic and moderately toxic substance used in DNA-staining during electrophoresis. Interest in phytoremediation as a method to solve chemical contamination has been growing rapidly in recent years. The technology has been utilized to clean up soil and groundwater from heavy metals and other toxic organic compounds in many countries like the United States, Russia, and most of European countries. Phytoremediation requires somewhat limited resources and very useful in treating wide variety of environmental contaminants. This study aimed to assess the potential of selected tropical plants as phytoremediators of EtBr. Materials and Methods This study used tomato (Solanum lycopersicum), mustard (Brassica alba), vetivergrass (Vetiveria zizanioedes), cogongrass (Imperata cylindrica), carabaograss (Paspalum conjugatum), and talahib (Saccharum spontaneum) to remove EtBr from laboratory wastes. The six tropical plants were planted in individual plastic bags containing soil and 10% EtBr-stained agarose gel. The plants were allowed to establish and grow in soil for 30 days. Ethidium bromide content of the test plants and the soil were analyzed before and after soil treatment. Ethidium bromide contents of the plants and soils were analyzed using an UV VIS spectrophotometer. Results Results showed a highly significant (p≤0.001) difference in the ability of the tropical plants to absorb EtBr from soils. Mustard registered the highest absorption of EtBr (1.4±0.12 μg kg⁻¹) followed by tomato and vetivergrass with average uptake of 1.0±0.23 and 0.7±0.17 μg kg⁻¹ EtBr, respectively. Cogongrass, talahib, and carabaograss had the least amount of EtBr absorbed (0.2±0.6 μg kg⁻¹). Ethidium bromide content of soil planted to mustard was reduced by 10.7%. This was followed by tomato with an average reduction of 8.1%. Only 5.6% reduction was obtained from soils planted to vetivergrass. Soils planted to cogongrass, talahib, and carabaograss had the least reduction of 1.52% from its initial EtBr content. Discussion In this study, mustard, tomato, and vetivergrass have shown their ability to absorb EtBr from contaminated soil keeping them from expanding their reach into the environment and preventing further contamination. Its downside, however, is that living creatures including humans, fish, and birds, must be prevented from eating the plants that utilized these substances. Nonetheless, it is still easier to isolate, cut down, and remove plants growing on the surface of the contaminated matrices, than to use strong acids and permanganates to chemically neutralize a dangerous process that can further contaminate the environment and pose additional risks to humans. Though this alternative method does not totally eliminate eventual environmental contamination, it is by far produces extremely insignificant amount of by-products compared with the existing processes and technologies. Conclusions Mustard had the highest potential as phytoremediator of EtBr in soil. However, the absorption capabilities of the other test plants may also be considered in terms of period of maturity and productivity. Recommendations and Perspectives It is recommended that a more detailed and complete investigation of the phytoremediation properties of the different plants tested should be conducted in actual field experiments. Plants should be exposed until they reach maturity to establish their maximum response to the toxicity and mutagenecity of EtBr and their maximum absorbing capabilities. Different plant parts should be analyzed individually to determine the movement and translocation of EtBr from soil to the tissues of plants. Since this study has established that some plants can thrive and dwell in EtBr-treated soil, an increased amount of EtBr application should be explored in future studies. It is suggested therefore that a larger, more comprehensive exploration of phytoremediation application in the management of toxic and hazardous wastes emanating from biotechnology research activities should be considered especially on the use of vetivergrass, a very promising tropical perennial grass. ESS-Submission Editor: Professor Zhihong Xu (zhihong.xu@griffith.edu.au: www.griffith.edu.au/centre/cfhr)     

Wastewater treatment plants and landfills emit volatile methyl siloxanes (VMSs) to the atmosphere: investigations using a new passive air sampler

Volatile methyl siloxanes (VMSs) are a class of chemicals with an increasing range of applications and widespread distribution in the environment. Passive air samplers (PAS) comprising sorbent-impregnated polyurethane-foam (SIP) disks were first calibrated and then deployed around two wastewater treatment plants (WWTPs) and at two landfill sites to investigate inputs of VMSs to air. SIP-derived air concentrations for ΣVMSs (ng/m³) at background sites on the perimeter of the WWTP were 479 ± 82.3 and comparable to results for the upwind samples at the landfills (333 ± 194). Order of magnitude higher concentrations of ΣVMSs (ng/m³) were found for on-site samples at the WWTPs (3980 ± 2620) and landfills (4670 ± 3360). Yearly emissions (kg/yr) to air were estimated and ranged from 60-2100 and 80-250, respectively, for the cyclic VMSs. These findings demonstrate and quantify for the first time, waste sector emissions of VMSs to the atmosphere.