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.     

Volatile sesquiterpenes from Stachybotrys chartarum: Indicators for trichothecene producing mold species?

Trichodiene, a volatile sesquiterpene which is structurally related to trichothecene mycotoxins, has been identified in the headspace of growing Stachybotrys chartarum by GC/MS. It is possible that volatile sesquiterpene patterns can be used to characterize S. chartarum and related mold isolates as trichothecene producers, thus providing clear criteria for decisions concerning the occupancy and renovation of contaminated buildings.     

Effects of lindane on the photosynthetic apparatus of the cyanobacterium<Emphasis Type="Italic">anabaena</Emphasis>

INTENTION, GOAL, SCOPE, BACKGROUND: Cyanobacteria have the natural ability to degrade moderate amounts of organic pollutants. However, when pollutant concentration exceeds the level of tolerance, bleaching of the cells and death occur within 24 hours. Under stress conditions, cyanobacterial response includes the short-term adaptation of the photosynthetic apparatus to light quality, named state transitions. Moreover, prolonged stresses produce changes in the functional organization of phycobilisomes and in the core-complexes of both photosystems, which can result in large changes in the PS II fluorescence yield. The localization of ferredoxin-NADP+ reductase (FNR) at the ends of some peripheral rods of the cyanobacterial phycobilisomes, makes this protein a useful marker to check phycobilisome integrity. OBJECTIVE: The goal of this work is to improve the knowledge of the mechanism of action of a very potent pesticide, lindane (gamma-hexaclorociclohexane), in the cyanobacterium Anabaena sp., which can be considered a potential candidate for bioremediation of pesticides. We have studied the effect of lindane on the photosynthetic apparatus of Anabaena using fluorescence induction studies. As ferredoxin-NADP+ reductase plays a key role in the response to oxidative stress in several systems, changes in synthesis, degradation and activity of FNR were analyzed. Immunolocalization of this enzyme was used as a marker of phycobilisome integrity. The knowledge of the changes caused by lindane in the photosynthetic apparatus is essential for rational further design of genetically-modified cyanobacteria with improved biorremediation abilities. METHODS: Polyphasic chlorophyll a fluorescence rise measurements (OJIP) have been used to evaluate the vitality and stress adaptation of the nitrogen-fixing cyanobacterium Anabaena PCC 7119 in the presence of increasing concentrations of lindane. Effects of the pesticide on the ultrastructure have been investigated by electron microscopy, and FNR has been used as a marker of phycobilisome integrity. RESULTS AND DISCUSSION: Cultures of Anabaena sp. treated with moderate amounts of lindane showed a decrease in growth rate followed by a recovery after 72 hours of pesticide treatment. Concentrations of lindane below 5 ppm increased the photosynthetic performance and activity of the cells. Higher amounts of pesticide caused a decrease in these activities which seems to be due to a non-competitive inhibition of PS II. Active PS II units are converted into non-QA reducing, so called heat sink centers. Specific activity and amount of FNR in lindane-treated cells were similar to the values measured in control cultures. Release of FNR from the thylakoid after 48 hours of exposure to 5 ppm of lindane towards the cytoplasm was detected by immunogold labeling and electron microscopy. CONCLUSIONS: From these results, we conclude that the photosynthetic performance and activity of the cells are slightly increased in the presence of lindane up to 5 ppm. Moreover, in those conditions, lindane did not produce significant changes in the synthesis, degradation or activity of FNR. The high capability of Anabaena to tolerate lindane makes this cyanobacterium a good candidate for phytoremediation of polluted areas. RECOMMENDATION AND OUTLOOK: The results of this study show that cultures of Anabaena PCC 7119 tolerate lindane up to 5 ppm, without significant changes in the photosynthetic vitality index of the cells. However, a slight increase in phycobiliprotein synthesis is observed, which is related to total protein content. This change might be due to degradation of proteins less stable than phycobiliproteins. An identification of the proteins with altered expression pattern in the presence of the pesticide remains the subject of further work and will provide valuable information for the preparation of strains which are highly tolerant to lindane.     

The effect of 4-Nonylphenol on the pigmentation of<Emphasis Type="Italic">Ocimum basilicum</Emphasis> (Basil)

BACKGROUND, AIM AND SCOPE: Tests during the last few years have confirmed that 4-nonylphenol (4-NP) can have oestrogen-like effects (xeno-hormone) on animal organisms. The objective was to firstly evaluate the ecotoxicological effects of 4-NP on plants such as hydrocultures. To clarify how this substance interferes with the photosynthetic system of plants, various tests were carried out using the basil plant (Ocimum basilicum). METHODS: The effect of the pollutant 4-NP on the pigment content in the leaves of the basil plant was analysed with the use of High-Performance-Liquid-Chromatography (HPLC). RESULTS AND DISCUSSION: A general assessment of the HPLC data revealed that plants that came in contact with the 4-nonylphenol showed a change in pigmentation. More chlorophyll a and b was produced, although at the same time a higher production of degradation products and by-products of the chlorophylls was observed. These occurrences can therefore be seen as an impairment of the photosynthetic process. The contaminated plants produced less xanthophylls than the non-contaminated ones, though these differences were statistically not significant. CONCLUSION AND PERSPECTIVE: The variations on the pigment content in the leaves of the basil plant can be interpreted as a consequence of the 4-NP application. It was, however, not investigated whether the plants absorbed the pollutants directly. The effect could have been caused by adsorption of the oily substance to the roots, and this could have led to a hindrance of the uptake of nutrients and possibly water. In order to clarify this further, biochemical experiments are being conducted.     

The role of the egg jelly coat in protecting<Emphasis Type="Italic">Hyla regilla</Emphasis> and<Emphasis Type="Italic">Bufo canorus</Emphasis> embryos from ultraviolet B radiation during development

BACKGROUND: Previous studies have suggested that Ultraviolet B (UVB) radiation may play a role in amphibian population declines. Some of these studies also indicate that egg hatching success is unaltered in some species of anurans as a result of UVB exposure. It has been proposed that the egg mass jelly provides photoprotection to the developing embryos. METHODS: Direct spectrophotometric scans of egg jelly, scans of egg jelly methanol extracts, and experimental manipulation in a solar simulator during development were all used to assess the role of egg mass jelly as a photoprotective agent. RESULTS/DISCUSSION: For Hyla regilla, scans of egg jelly and methanolic extracts (for mycosporine-like amino acid content) both displayed no absorption in the UV range. Experimental manipulation (removal of egg mass jelly) with both Hyla regilla and Bufo canorus egg masses in a solar simulator demonstrated that egg mass jelly played no apparent role in photoprotection of either of these species. CONCLUSIONS: Based on the results in this study it seems unlikely that the egg jelly coat is playing a crucial role in protecting developing embryos from the impact of UVB radiation.     

Toxicity of cobalt-complexed cyanide to <Emphasis Type="Italic">Oncorhynchus mykiss,Daphnia magna</Emphasis>, and <Emphasis Type="Italic">Ceriodaphnia dubia</Emphasis>

Background Cobalt cyanide complexes often result when ore is treated with cyanide solutions to extract gold and other metals. These have recently been discovered in low but significant concentrations in effluents from gold leach operations. This study was conducted to determine the potential toxicity of cobalt-cyanide complexes to freshwater organisms and the extent to which ultraviolet radiation (UV) potentiates this toxicity. Tests were also conducted to determine if humic acids or if adaptation to UV influenced sensitivity to the cyanide complexes. Methods Rainbow trout (Oncorhynchus mykiss), Daphnia magna, and Ceriodaphnia dubia were exposed to potassium hexacyanocobaltate in the presence and absence of UV radiation, in the presence and absence of humic acids. Cyano-cobalt exposures were also conducted with C. dubia from cultures adapted to elevated UV. Results With an LC50 concentration of 0.38 mg/L, cyanocobalt was over a 1000 times more toxic to rainbow trout in the presence of UV at a low, environmentally relevant irradiance level (4 μW/cm² as UVB) than exposure to this compound in the absence of UV with an LC50 of 112.9 mg/L. Toxicity was immediately apparent, with mortality occurring within an hour of the onset of exposure at the highest concentration. Fish were unaffected by exposure to UV alone. Weak-acid dissociable cyanide concentrations were observed in irradiated aqueous solutions of cyanocobaltate within hours of UV exposure and persisted in the presence of UV for at least 96 hours, whereas negligible concentrations were observed in the absence of UV. The presence of humic acids significantly diminished cyanocobalt toxicity to D. magna and reduced mortality from UV exposure. Humic acids did not significantly influence survival among C. dubia. C. dubia from UV-adapted populations were less sensitive to metallocyanide compounds than organisms from unadapted populations. Conclusions The results indicate that metallocyanide complexes may pose a hazard to aquatic life through photochemically induced processes. Factors that decrease UV exposure such as dissolved organic carbon or increased pigmentation would diminish toxicity.     

Impact of biogenic terpene emissions from <Emphasis Type="Italic">Brassica napus</Emphasis> on tropospheric ozone over saxony (Germany)

INTRODUCTION: The role of biogenic emissions in tropospheric ozone production is currently under discussion and major aspects are not well understood yet. This study aims towards the estimation of the influence of biogenic emissions on tropospheric ozone concentrations over Saxony in general and of biogenic emissions from brassica napus in special. MODELLING TOOLS: The studies are performed by utilizing a coupled numerical modelling system consisting of the meteorological model METRAS and the chemistry transport model MUSCAT. For the chemical part, the Euro-RADM algorithm is used. EMISSIONS: Anthropogenic and biogenic emissions are taken into account. The anthropogenic emissions are introduced by an emission inventory. Biogenic emissions, VOC and NO, are calculated within the chemical transport model MUSCAT at each time step and in each grid cell depending on land use type and on the temperature. The emissions of hydrocarbons from forest areas as well as biogenic NO especially from agricultural grounds are considered. Also terpene emissions from brassica napus fields are estimated. SIMULATION SETUP AND METEOROLOGICAL CONDITIONS: The simulations were performed over an area with an extension of 160 x 140 km2 which covers the main parts of Saxony and neighboring areas of Brandenburg, Sachsen-Anhalt and Thuringia. Summer smog with high ozone concentrations can be expected during high pressure conditions on hot summer days. Typical meteorological conditions for such cases were introduced in an conceptual way. RESULTS: It is estimated that biogenic emissions change tropospheric ozone concentrations in a noticeable way (up to 15% to 20%) and, therefore, should not be neglected in studies about tropospheric ozone. Emissions from brassica napus do have a moderate potential to enhance tropospheric ozone concentrations, but emissions are still under consideration and, therefore, results vary to a high degree. CONCLUSIONS: Summing up, the effect of brassica napus terpene emissions on ozone concentrations is noticeable, but not too pronounced. The results give a preliminary estimate on what the effect due to brassica napus emissions could be until better parameterizations can be derived from measurements.     

Toxic effects of orimulsion on rainbow trout <Emphasis Type="Italic">oncorhynchus mykiss</Emphasis>

GOAL, SCOPE AND BACKGROUND: Orimulsion (stable emulsion of natural bitumen and water) is a new imported industrial fuel in Lithuania. No data on its toxicity to fish is freely available. The aim of this study was to investigate sensitivity of rainbow trout (Oncorhynchus mykiss) to acute and chronic toxicity of orimulsion and to estimate the Maximum Acceptable Toxicant Concentration (MATC) of orimulsion to fish. METHODS: Laboratory tests were conducted on rainbow trout in all stages of development (embryos, larvae, adults). Acute toxicity (96-hour duration) and long-term (28 or 60-day duration) tests evaluating the wide range spectrum of biological indices were performed under semi-static conditions. RESULTS AND DISCUSSION: Median lethal concentration (96-hour LC50) values and their 95% confidence intervals derived from the tests were: 0.1 (0.09-0.12) to embryos, 0.06 (0.05-0.07) to larvae and 2.22 (2.02-2.43) to adult fish, and 28-day LC50 to adult fish was found to be 0.26 (0.21-0.32) g/l of total orimulsion respectively. The acute toxicity of orimulsion to rainbow trout can be characterised by a narrow zone of toxic effect and a sharp boundary between lethal and sublethal concentrations. The lowest 'safe' or 'no-effect' concentration values of total orimulsion obtained in long-term tests were equal to 0.09 g/l to adult fish, 0.019 g/l to embryos, and 0.0017 g/l to larvae. Proposed value of 'application factor' for orimulsion was found to be equal to 0.03. Since orimulsion has the property to disperse in all water volume, its toxic effect on fish can be characterised by the combined effects of dispersion and water-soluble-fraction. CONCLUSIONS: Maximum Acceptable Toxicant Concentration (MATC) of 0.0017 g/l of total orimulsion to fish was derived from long-term tests based on the most sensitive parameter of rainbow trout larvae (relative mass increase at the end of the test). According to substance toxicity classification accepted for Lithuanian inland waters, orimulsion can be referred to substances of 'moderate' toxicity to fish. RECOMMENDATIONS AND OUTLOOK: For prediction and evaluation of toxic impact of orimulsion accident spills on fish, some recommendations should be given. Since orimulsion has the property to disperse in all water volume during short time periods, the amounts of both spilled orimulsion and polluted water should be ascertained. Once both parameters are known, the real concentration of orimulsion in the water body must be determined. Then this concentration must be compared with 'safe' concentration to fish. By use of 'application factor' 0.03, approximate MATC for other fish species can be estimated when only acute toxicity data (96-hour LC50 value) is available.     

Effect of a heavy metal model mixture on biological parameters of rainbow trout <Emphasis Type="Italic">oncorhynchus mykiss</Emphasis>

The effects of a model mixture (HMMM) of seven heavy metals (Cu, Zn, Ni, Cr, Pb, Cd, Mn) on the rainbow trout Oncorhynchus mykiss at all stages of development (embryos, larvae, adults) were investigated based on the annual average concentrations of these metals in cooling waste waters discharging from Ignalina Nuclear Power Plant (Lithuania) into the Drūk?iai lake. According to mortality parameters, the most sensitive to HMMM were larvae, although no significant differences between the sensitivity of embryos and adult fish to HMMM were found. Maximal toxic effect of HMMM was observed during the hatching period. Long-term exposure to sublethal concentrations of HMMM affected embryo development, growth of larvae, their cardio-respiratory and behavioural responses, induced significant changes in morphological, morpho-physiological, physiological and haematological parameters of adult fish. Respiratory responses and growth parameters of fish were found to be the most sensitive to low concentrations of HMMM. Adult fish were capable of detecting and avoiding low, sublethal concentrations of HMMM. Heavy metals in a mixture at low concentrations were more toxic than single ones. According to the background of the damages induced by HMMM, after-effects in a fish organism, as well as in a whole population, can be predicted.     

Testing antimicrobial cleaner efficacy on gypsum wallboard contaminated with <Emphasis Type="Italic">Stachybotrys chartarum</Emphasis>

Goal, Scope and Background Reducing occupant exposure to indoor mold is the goal of this research, through the efficacy testing of antimicrobial cleaners. Often mold contaminated building materials are not properly removed, but instead surface cleaners are applied in an attempt to alleviate the problem. The efficacy of antimicrobial cleaners to remove, eliminate or control mold growth on surfaces can easily be tested on non-porous surfaces. However, the testing of antimicrobial cleaner efficacy on porous surfaces, such as those found in the indoor environment such as gypsum board can be more complicated and prone to incorrect conclusions regarding residual organisms. The mold Stachybotrys chartarum has been found to be associated with idiopathic pulmonary hemorrhage in infants and has been studied for toxin production and its occurrence in water damaged buildings. Growth of S. chartarum on building materials such as gypsum wallboard has been frequently documented. Methods Research to control S. chartarum growth using 13 separate antimicrobial cleaners on contaminated gypsum wallboard has been performed in laboratory testing. Popular brands of cleaning products were tested by following directions printed on the product packaging. Results A variety of gypsum wallboard surfaces were used to test these cleaning products at high relative humidity. The results indicate differences in antimicrobial efficacy for the six month period of testing. Discussion Results for the six types of GWB surfaces varied extensively. However, three cleaning products exhibited significantly better results than others. Lysol All-Purpose Cleaner-Orange Breeze (full strength) demonstrated results which ranked among the best in five of the six surfaces tested. Both Borax and Orange Glo Multipurpose Degreaser demonstrated results which ranked among the best in four of the six surfaces tested. Conclusions The best antimicrobial cleaner to choose is often dependent on the type of surface to be cleaned of S. chartarum contamination. For Plain GWB, no paint, the best cleaners were Borax, Lysol All-Purpose Cleaner-Orange Breeze (full strength), Orange Glo Multipurpose Degreaser, and Fantastik Orange Action. Recommendations and Perspectives These results are not meant to endorse the incomplete removal of mold contaminated building materials. However, it is recognized that complete removal may not always be possible and solutions to control mold regrowth may contribute to reduced occupant exposure. Current recommendations of removal and replacement of porous building materials should be followed. It is not the intension of this discussion to endorse any product. Reporting on the performance of these products under the stated conditions was and remains the only purpose. ESS-Submission Editor: Dr. Lee Young (youngrisk@bresnan.net)     

Bactericidal activity of Cu-, Zn-, and Ag-containing zeolites toward <Emphasis Type="Italic">Escherichia coli</Emphasis> isolates

Two types of zeolites—natural clinoptilolite (NZ) and synthetic zeolite A (A)—were enriched with approx. 0.25 mmol of Cu(II), Zn(II), or Ag(I) ions, and the obtained materials (M-Z) were tested against three different isolates of Escherichia coli. Two isolates were environmental isolates from waters in Serbia whereas the third one was DSM 498. Antibacterial activity was studied in different water media—nutrient-rich media (peptone water), water from Sava Lake, and commercially available spring water. The Ag-containing zeolites showed bactericidal activity in the nutrient-rich peptone water after 1 h of contact. Cu- and Zn-containing zeolites showed bactericidal activity in real water samples. Antibacterial activity of the M-Z decreases in all three examined water media in the following order: Ag-NZ ≈ Ag-A > Cu-NZ ≈ Cu-A > Zn-NZ >>> Zn-A, suggesting that mainly the metal type and not the zeolite type have a role in the antibacterial activity. Leaching experiments showed small amounts of the leached Cu(II) and Zn(II) ions, indicating that the antibacterial activity is not due to the metal ions but should be attributed to the M-Z itself. However, leached amounts of Ag(I) from Ag-NZ and Ag-A in peptone water indicate that the released Ag(I) could be mainly responsible for the bactericidal effect of the Ag(I)-containing zeolites. Since no loss of cellular material was found, the antibacterial activity is not attributed to cytoplasmic membrane damage.     

Chlorinated dibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxins and dibenzofurans in the baltic herrinj and sprat of estonian coastal waters

BACKGROUND, AIMS AND SCOPE: The concentration of chlorinated dibenzo-p-dioxins and dibenzofurans in many fish from the Baltic requires monitoring, since it approaches or exceeds the European Union threshold limit value of 4 pg TEQ/g wet weight of fish for human consumption. The concentrations, expressed in TEQs, are important for toxicology and regulations, but hide the concentrations of the individual congeners, which are important for other environmental sciences, source allocation, and for the detection of measurement errors. This report evaluates the results of a survey reported earlier only in the terms of the TEQ concentrations. METHODS: Principal Component Analysis (PCA) was used to reduce the dimensions of the data (17 = 7 chlorinated dibenzo-p-dioxin and 10 chlorinated dibenzofuran congeners) to three principal components. This facilitated the interpretation of the congener profiles. Slopes of the congener concentrations as a function of age of the fish were estimated by least squares regression. The results were compared with a large set of data for lake trout from Lake Ontario. RESULTS AND DISCUSSION: The congener profiles of Baltic herring are less scattered than those of sprat. The profiles of herring from the central Baltic differ from those of herring from the Gulf of Riga and both appear to be affected relatively minimally by the age of the fish. The congener profiles of herring from the western Gulf of Finland are similar to those from the central Baltic, those from middle Gulf of Finland are similar to those from the Gulf of Riga. Both seem to be more affected by age of the fish than the profiles of the first two groups. The concentrations of several pentachloro- and hexachloro-dibenzo-p-dioxins and dibenzofurans increase with the age of the fish CONCLUSION: PCA is a good technique for the evaluation of the congener profiles. The resulting loading and score plots provide a good graphic summary of the multidimensional data. Additional analyses are needed to confirm the observed profile patterns. A comparison with the results of a long-term monitoring from another area shows that the age of the fish is a more important factor than the year of capture. RECOMMENDATION AND OUTLOOK: The surveys should continue for a number of years and the results should be presented and evaluated both in terms of the TEQs as well as in terms of weight concentrations. Since the concentrations do not appear to change very much from year to year, it would be better to carry out surveys only every 3-4 years and, instead, stratify the sampling according to age and gender of the fish, and to analyze replicate extracts by replicate measurements. The inclusion of unmarked replicate samples would be a good quality assurance measure. It would be desirable to analyze additional parts of the food chain in order to understand the fate of the compounds in the ecosystem.     

Uptake and degradation of trimethylamine by <Emphasis Type="Italic">Euphorbia milii</Emphasis>

Trimethylamine (TMA) is a volatile organic compound which causes not only unpleasant odor but also health concerns to humans. The average emission of TMA from food and fishery industries is 20.60 parts per billion (ppb) and emission from the gas exhausters is even higher which reaches 370 parts per million (ppm). In order to select the best plant TMA removal agent, in this study, 13 plants were exposed to 100 ppm of TMA and the remaining TMA concentration in their system was analyzed by gas chromatography (GC). Furthermore, plant metabolites from the selected plant were identified by gas chromatography-mass spectrometry (GC-MS). The result showed that Euphorbia milii was the most superior plant for TMA removal and could absorb up to 90 % of TMA within 12 h. E. milii absorbed TMA via leaf and stem with 55 and 45 % uptake efficiency, respectively. Based on its stomatal movement during the exposure to TMA, it was implied that the plant switched the photosynthetic mode from crassulacean acid metabolism (CAM)-cycling to CAM and CAM-idling. The switching of photosynthetic mode might reduce the stomata role in TMA absorption. Fatty acids, alkanes, and fatty alcohols in the plant leaf wax were also found to contribute to TMA adsorption. Leaf wax, stomata, and other leaf constituents contributed 58, 6, and 36 %, respectively, of the total TMA absorption by the leaf. The analysis and identification of plant metabolites confirmed that TMA was degraded and mineralized by E. milii.     

Exposure of the Roots of <Emphasis Type="Italic">Populus nigra</Emphasis> L. cv. Loenen to PAHs and its Effect on growth and water balance

Poplar cuttings were cultivated for 4 weeks in a substrate, which consisted of a combination of sand and nutrient solution. The plants were treated for 24 days with BaP, Chr, Ant, Phen, P and Flt, single or in combination. The concentration of the PAHs ranged from 0.1-200 mg/kg substrate. The results of the pollution experiments can be summarized as follows: 1. The most significant deviations between the test groups and the control can be observed for transpiration, nutrient solution uptake, and root mass. 2. Although transpiration and nutrient solution uptake are significantly lower for all the treated groups than for the control group, the water content of the leaves was not affected by PAHs. 3. The biomass of the shoots and the growth in shoot length do not react as strongly to exposure to PAHs as transpiration, nutrient solution uptake and the volume of the roots. 4. The differences in leaf weight and leaf surface area are significantly less pronounced compared to the control groups. Growth inhibition is most evident with Flt. Growth and absorption of the nutrient solution dropped with just Flt 0.1 mg/kg substrate. When the substrate concentration was increased, growth and nutrient solution uptake dropped considerably and at a concentration of Flt 200, 5 of the 11 test plants died before the end of the period of exposure. Nutrient solution uptake and shoot development of the test plants decreased in the following order: BaP H approximately = Chr > Ant > Phen > Pyr > Flt.     

Phytoremediation Potential of Aquatic Macrophyte, <Emphasis Type="Italic">Azolla</Emphasis>

Aquatic macrophytes play an important role in the structural and functional aspects of aquatic ecosystems by altering water movement regimes, providing shelter to fish and aquatic invertebrates, serving as a food source, and altering water quality by regulating oxygen balance, nutrient cycles, and accumulating heavy metals. The ability to hyperaccumulate heavy metals makes them interesting research candidates, especially for the treatment of industrial effluents and sewage waste water. The use of aquatic macrophytes, such as Azolla with hyper accumulating ability is known to be an environmentally friendly option to restore polluted aquatic resources. The present review highlights the phytoaccumulation potential of macrophytes with emphasis on utilization of Azolla as a promising candidate for phytoremediation. The impact of uptake of heavy metals on morphology and metabolic processes of Azolla has also been discussed for a better understanding and utilization of this symbiotic association in the field of phytoremediation.     

Combined biocidal action of silver nanoparticles and ions against Chlorococcales (S<Emphasis Type="Italic">cenedesmus quadricauda</Emphasis>, <Emphasis Type="Italic">Chlorella vulgaris)</Emphasis> and filamentous algae (<Emphasis Type="Italic">Klebsormidium sp.</Emphasis>)

Despite the extensive research, the mechanism of the antimicrobial and biocidal performance of silver nanoparticles has not been unequivocally elucidated yet. Our study was aimed at the investigation of the ability of silver nanoparticles to suppress the growth of three types of algae colonizing the wetted surfaces or submerged objects and the mechanism of their action. Silver nanoparticles exhibited a substantial toxicity towards Chlorococcales Scenedesmus quadricauda, Chlorella vulgaris, and filamentous algae Klebsormidium sp., which correlated with their particle size. The particles had very good stability against agglomeration even in the presence of multivalent cations. The concentration of silver ions in equilibrium with nanoparticles markedly depended on the particle size, achieving about 6 % and as low as about 0.1 % or even less for the particles 5 nm in size and for larger ones (40–70 nm), respectively. Even very limited proportion of small particles together with larger ones could substantially increase concentration of Ag ions in solution. The highest toxicity was found for the 5-nm-sized particles, being the smallest ones in this study. Their toxicity was even higher than that of silver ions at the same silver concentration. When compared as a function of the Ag⁺ concentration in equilibrium with 5-nm particles, the toxicity of ions was at least 17 times higher than that obtained by dissolving silver nitrite (if not taking into account the effect of nanoparticles themselves). The mechanism of the toxicity of silver nanoparticles was found complex with an important role played by the adsorption of silver nanoparticles and the ions released from the particles on the cell surface. This mechanism could be described as some sort of synergy between nanoparticles and ions. While our study clearly showed the presence of this synergy, its detailed explanation is experimentally highly demanding, requiring a close cooperation between materials scientists, physical chemists, and biologists.     

Comparing growth development of <Emphasis Type="Italic">Myriophyllum</Emphasis> spp. in laboratory and field experiments for ecotoxicological testing

BACKGROUND, GOALS AND SCOPE: Risk assessment of herbicides and the evaluation of contaminated sediments based on algae and the macrophyte Lemna sp. alone may underestimate the potential hazard of certain compounds. Therefore, various test systems with Myriophyllum spp. have been developed recently to assess the phytotoxicity in surface waters and natural sediments. In the present study, experiments investigating the growth development of Myriophyllum spp. were performed in the laboratory under defined conditions and in mesocosms under environmentally realistic exposure conditions to evaluate the suitability of these species as potential standard test organisms in ecotoxicological testing. This study provides data on the endpoints biomass, plant length and root development. MATERIALS AND METHODS: Six independent experiments were performed to investigate the plant development of Myriophyllum spp. under control conditions. The main difference in the experiments was the complexity of the test systems ranging from simple laboratory experiments to complex outdoor mesocosm studies. At the start of each experiment, uniform cuttings of Myriophyllum spp. were placed in vessels with or without sediments to reduce variability between replicates. The endpoints considered in this investigation were biomass (fresh weight of the whole plant), length of the main shoot, length of the side shoots, total length of the plant (calculated from the length of the main and side shoots) and root formation. Root to shoot ratios were calculated as a further measure for plant development. Relative growth rates (RGR) based on plant length (RG(L)R) and on biomass (RG(B)R) were calculated. RESULTS: Despite the various experimental conditions, comparable growth was obtained in all test systems and the variability of endpoints, such as total length and biomass of plants, was low. It was observed that the RGR of M. spicatum in the simple laboratory test system with sediment were comparable to growth data obtained for M. verticillatum and M. spicatum grown in indoor and outdoor mesocosms, thus indicating that Myriophyllum growth tends to increase by the addition of sediment. High variability was determined for the endpoints length of the side shoots, total root length and biomass of roots. DISCUSSION: One challenge for a test design to investigate phytotoxicity on aquatic plants is to obtain good growth of the plants. From the results, it can be concluded that the experimental conditions in the various test systems were suitable to study the plant development of Myriophyllum spp. because obtained growth rates were comparable between laboratory and field investigations. Another challenge for developing a plant biotest system is the definition of sensitive endpoints. Low variability is preferred to detect minor effects of chemicals or polluted sediments on plant development. In our studies, the variability of the endpoints biomass and total length of plant was low and, therefore, they have much potential as endpoints for assessing toxicity. CONCLUSIONS: The methodologies presented in this study have applications within the risk assessment for aquatic plants and have the advantage of assessing effects taking into account the relevant exposure pathways via water and/or sediment for compounds under investigation. RECOMMENDATIONS AND PERSPECTIVES: Setting safe quality criteria for surface water and sediments is one of the challenges authorities are facing today. Myriophyllum spp. is recommended as suitable test species to investigate phytotoxicity in surface water and sediments. These results, thus, might serve as a basis for the compilation of a new harmonised guideline for ecotoxicological testing with aquatic macrophytes.     

Selected endocrine disrupting compounds (Vinclozolin,Flutamide, Ketoconazole and Dicofol): Effects on survival,occurrence of males,growth, molting and reproduction of <Emphasis Type="Italic">Daphnia magna</Emphasis>

BACKGROUND, AIM AND SCOPE: Pollution-induced endocrine disruption in vertebrates and invertebrates is a worldwide environmental problem, but relatively little is known about effects of endocrine disrupting compounds (EDCs) in planktonic crustaceans (including Daphnia magna). Aims of the present study were to investigate acute 48 h toxicity and sub-chronic (4-6 days) and chronic (21 days) effects of selected EDCs in D. magna. We have investigated both traditional endpoints as well as other parameters such as sex determination, maturation, molting or embryogenesis in order to evaluate the sensitivity and possible use of these endpoints in ecological risk assessment. MATERIALS AND METHODS: We have studied effects of four model EDCs (vinclozolin, flutamide, ketoconazole and dicofol) on D. magna using (i) an acute 48 h immobilization assay, (ii) a sub-chronic, 4-6 day assay evaluating development and the sex ratio of neonates, and (iii) a chronic, 21 day assay studying number of neonates, sex of neonates, molting frequency, day of maturation and the growth of maternal organisms. RESULTS: Acute EC50 values in the 48 h immobilization test were as follows (mg/L): dicofol 0.2, ketoconazole 1.5, flutamide 2.7, vinclozolin >3. Short-term, 4-6 day assays with sublethal concentrations showed that the sex ratio in Daphnia was modulated by vinclozolin (decreased number of neonate males at 1 mg/L) and dicofol (increase in males at 0.1 mg/L). Flutamide (up to 1 mg/L) had no effect on the sex of neonates, but inhibited embryonic development at certain stages during chronic assay, resulting in abortions. Ketoconazole had no significant effects on the studied processes up to 1 mg/L. DISCUSSION: Sex ratio modulations by some chemicals (vinclozolin and dicofol) corresponded to the known action of these compounds in vertebrates (i.e. anti-androgenicity and anti-oestrogenicity, respectively). Our study revealed that some chemicals known to affect steroid-regulated processes in vertebrates can also affect sublethal endpoints (e.g. embryonic sex determination and/or reproduction) in invertebrates such as D. magna. CONCLUSIONS: A series of model vertebrate endocrine disrupters affected various sub-chronic and chronic parameters in D. magna including several endpoints that have not been previously studied in detail (such as sex determination in neonates, embryogenesis, molting and maturation). Evaluations of traditional reproduction parameters (obtained from the 21 day chronic assay). as well as the results from a rapid, 4-6 day, sub-chronic assay provide complementary information on non-lethal effects of suspected organic endocrine disrupters. RECOMMENDATIONS AND PERSPECTIVES: It seems that there are analogies between vertebrates and invertebrates in toxicity mechanisms and in vivo effects of endocrine disruptors. However, general physiological status of organisms may also indirectly affect endpoints that are traditionally considered 'hormone regulated' (especially at higher effective concentrations as observed in this study) and these factors should be carefully considered. Further research of D. magna physiology and comparative studies with various EDCs will help to understand mechanisms of action as well as ecological risks of EDCs in the environment.     

Endpoint ‘floating leaves’ of <Emphasis Type="Italic">Potamogeton natans</Emphasis>: A new method to evaluate the development of macrophytes in pond mesocosms

Goal, Scope and Background One of the advantages of long-term mesocosm experiments as compared to short-term standard toxicity tests in the laboratory is the potential for detecting secondary effects due to the interaction of species and recovery with biomass of macrophytes being an important endpoint. However, generating biomass data by harvesting is often laborious, time-consuming, costly and restricted to the end of the experiment. Moreover, valuable information may get lost, in particular in single application studies, since maximal primary effects and secondary effects or recovery occur per se at different times. Potamogeton natans was used as an example in order to test whether number and area of floating leaves can be reliably measured and be used as intermediate and final endpoints in mesocosm effect studies. Methods Digital photos, which were taken of the water surface in the course of an indoor pond mesocosm study on herbicide effects, were subjected to image analysis. The results were compared to wet weight and ash-free dry weight of Potamogeton at the end of the herbicide study. Results and Discussion Both number and area of floating leaves indicated the same herbicide effects as wet weight and ash-free dry weight of Potamogeton. Error introduced by the different work steps is small and can be further minimised by a number of method improvements. Recommendations and Perspectives In indoor mesocosm studies, errors due to the perspective adjustment may be circumvented by taking the photos perpendicular to the water surface. Correction for lens aberration, identical light conditions and the use of fluorescence images are considered promising. Field applications are proposed.