Background, Aim and Scope Numerous herbicides and xenobiotic organic pollutants are detoxified in plants to glutathione conjugates. Following this enzyme catalyzed reaction, xenobiotic GS-conjugates are thought to be compartmentalized in the vacuole of plant cells. In the present study, evidence is presented for long range transport of these conjugates in plants, rather than storage in the vacuole. To our knowledge this is the first report about the unidirectional long range transport of xenobiotic conjugates in plants and the exudation of a glutathione conjugate from the root tips. This could mean that plants possess an excretion system for unwanted compounds which give them similar advantages as animals. Materials and Methods: Barley plants (Hordeum vulgare L. cv. Cherie) were grown in Petri dishes soaked with tap water in the greenhouse. - Fluorescence Microscopy. Monobromo- and Monochlorobimane, two model xenobiotics that are conjugated rapidly in plant cells with glutathione, hereby forming fluorescent metabolites, were used as markers for our experiments. Their transport in the root could be followed sensitively with very good temporal and spatial resolution. Roots of barley seedlings were cut under water and the end at which xenobiotics were applied was fixed in an aperture with a thin latex foil and transferred into a drop of water on a cover slide. The cover slide was fixed in a measuring chamber on the stage of an inverse fluorescence microscope (Zeiss Axiovert 100). - Spectrometric enzyme assay. Glutathione S-transferase (GST) activity was determined in the protein extracts following established methods. Aliquots of the enzyme extract were incubated with 1-chloro-2,4-dinitrobenzene (CDNB), or monochlorobimane. Controls lacking enzyme or GSH were measured. - Pitman chamber experiments. Ten days old barley plants or detached roots were inserted into special incubation chambers, either complete with tips or decapitated, as well as 10 days old barley plants without root tips. Compartment A was filled with a transport medium and GSH conjugate or L-cysteine conjugate. Compartments B and C contained sugar free media. Samples were taken from the root tip containing compartment C and the amount of conjugate transported was determined spectro-photometrically. Results: The transport in roots is unidirectional towards the root tips and leads to exsudation of the conjugates at rates between 20 and 200 nmol min-1. The microscopic studies have been complemented by transport studies in small root chambers and spectroscopic quantification of dinitrobenzene-conjugates. The latter experiments confirm the microscopic studies. Furthermore it was shown that glutathione conjugates are transported at higher rates than cysteine conjugates, despite of their higher molecular weights. This observation points to the existence of glutathione specific carriers and a specific role of glutathione in the root. Discussion: It can be assumed that long distance transport of glutathione conjugates within the plant proceeds like GSH or amino acid transport in both, phloem and xylem. The high velocity of this translocation of the GS-X is indicative of an active transport. For free glutathione, a rapid transport-system is essential because an accumulation of GSH in the root tip inhibits further uptake of sulfur. Taking into account that all described MRP transporters and also the GSH plasmalemma ATPases have side activities for glutathione derivatives and conjugates, co-transport of these xenobiotic metabolites seems credible. - On the other hand, when GS-B was applied to the root tips from the outside, no significant uptake was observed. Thus it can be concluded that only those conjugates can be transported in the xylem which are formed inside the root apex. Having left the root once, there seems to be no return into the root vessels, probably because of a lack of inward directed transporters. Conclusions: Plants seem to possess the capability to store glutathione conjugates in the vacuole, but under certain conditions, these metabolites might also undergo long range transport, predominantly into the plant root. The transport seems dependent on specific carriers and is unidirectional, this means that xenobiotic conjugates from the rhizosphere are not taken up again. The exudation of xenobiotic metabolites offers an opportunity to avoid the accumulation of such compounds in the plant. Recommendations and Perspectives: The role of glutathione and glutathione related metabolites in the rhizosphere has not been studied in any detail, and only scattered data are available on interactions between the plant root and rhizosphere bacteria that encounter such conjugates. The final fate of these compounds in the root zone has also not been addressed so far. It will be interesting to study effects of the exuded metabolites on the biology of rhizosphere bacteria and fungi.

Background, Aim and Scope Polycyclic aromatic hydrocarbons (PAH) are important environmental contaminants which may lead to increased levels of neoplastic aberrations or tumours in fish liver. Therefore, monitoring of PAH and their effects are part of several international environmental programmes. The aim of the present field study was to investigate the concentrations of the PAH metabolites in fish bile, to elucidate spatial, seasonal and species differences as well as to discuss different strategies of normalisation with regard to environmental monitoring. Materials and Methods: PAH metabolites were determined in the bile fluid of dab (Limanda limanda) and flounder (Platichthys flesus) caught in the North Sea and Baltic Sea between 1997 and 2004. After enzymatic deconjugation, two metabolites were determined by means of HPLC. The limit of detection and the limit of quantification were calculated. The accuracy of the method was tested with a standard reference material. Results were referred to bile volume as well as to biliverdin. Results: The main metabolite, 1-hydroxypyrene, was determined in concentrations from <0.7 to 838 ng/ml in bile of dab (Limanda limanda) and flounder (Platichthys flesus) caught between 1997 and 2004. The values for 1-hydroxyphenanthrene in fish bile were considerably lower (<0.4 - 87 ng/ml). Significant differences in the 1-hydroxypyrene levels were found between summer and winter surveys as well as between the sampling sites in the data set from 2004 (383 dabs and 62 flounders): Highest levels of PAH contamination were found in dab from the German Bight and in flounder from the Baltic Sea. Discussion: Spatial differences in 1-hydroxypyrene concentrations between North Sea and Baltic Sea were discussed, as well as differences in relation to season, sex and species. Three parameters of normalisation (biliary protein, biliverdin and bile pigments) were discussed. Biliverdin was identified as a suitable parameter for the normalisation of PAH metabolites in field samples. Conclusions: Spatial differences in 1-hydroxypyrene concentrations of dab demonstrate the usefulness of PAH metabolites in fish bile as a monitoring parameter in marine regions. Significant differences in 1-hydroxypyrene concentrations were found between summer and winter sampling campaigns. This may be linked to an annual cycle of 1-hydroxyprene in dab. It is also possible that bile synthesis/release in dab differs between the seasons. There is no indication for a time trend from 1997 to 2004. Recommendations and Perspectives: It is recommended to relate PAH metabolites in fish bile to biliverdin concentrations. Although the concentrations are low in offshore regions and bile volumes are small, the method presented here allows one to measure PAH metabolites on an individual level which is a crucial prerequisite for meaningful monitoring studies.

GOAL, SCOPE AND BACKGROUND: Aquaculture activities are well known to be the major contributor to the increasing level of organic waste and toxic compounds in the aquaculture industry. Along with the development of intensive aquaculture in China, concerns are evoked about the possible effects of everincreasing aquaculture waste both on productivity inside the aquaculture system and on the ambient aquatic ecosystem. Therefore, it is apparent that appropriate waste treatment processes are needed for sustaining aquaculture development. This review aims at identifying the current status of aquaculture and aquaculture waste production in China. MAIN FEATURES: China is the world’s largest fishery nation in terms of total seafood production volume, a position it has maintained continuously since 1990. Freshwater aquaculture is a major part of the Chinese fishery industry. Marine aquaculture in China consists of both land-based and offshore aquaculture, with the latter mostly operated in shallow seas, mud flats and protected bays. The environmental impacts of aquaculture are also striking. RESULTS: Case studies on pollution hot spots caused by aquaculture have been introduced. The quality and quantity of waste from aquaculture depends mainly on culture system characteristics and the choice of species, but also on feed quality and management. Wastewater without treatment, if continuously discharged into the aquatic environment, could result in remarkable elevation of the total organic matter contents and cause considerable economy lost. Waste treatments can be mainly classified into three categories: physical, chemical and biological methods. DISCUSSION: The environmental impacts of different aquaculture species are not the same. New waste treatments are introduced as references for the potential development of the waste treatment system in China. The most appropriate waste treatment system for each site should be selected according to the sites’ conditions and financial status as well as by weighing the advantages and disadvantages of each system. Strategies and perspectives for sustainable aquaculture development are proposed, with the emphasis on environmental protection. CONCLUSIONS: Negative effects of waste from aquaculture to aquatic environment are increasingly recognized, though they were just a small proportion to land-based pollutants. Properly planned use of aquaculture waste alleviates water pollution problems and not only conserves valuable water resources but also takes advantage of the nutrients contained in effluent. It is highly demanding to develop sustainable aquaculture which keeps stocking density and pollution loadings under environmental capacity. RECOMMENDATIONS AND PERSPECTIVES: The traditional procedures for aquaculture waste treatment, mainly based on physical and chemical means, should be overcome by more site-specific approaches, taking into account the characteristics and resistibility of the aquatic environment. Further research needs to improve or optimize the current methods of wastewater treatment and reuse. Proposed new treatment technology should evaluate their feasibility at a larger scale for practical application.

Working group on effects of the convention on long-rangr transboundary air pollution

- DOI: http://dx.doi.org/10.1065/espr2006.04.299 Goal, Scope and Background This paper describes a statistical modelling approach, suggested as a policy tool in the Athens area for the assessment of the emissions reduction level required to meet the air quality standards for two criteria air pollutants, namely CO and NO2. Methods. More than ten years of hourly CO and NOx-NO2 concentration data measured by the monitoring network of the Hellenic Ministry for the Environment, Physical Planning and Public Works were analyzed and the original dataset has been reduced using a data evaluation procedure. Results and Discussion Seasonal pollutant concentration trends suggested that the reduction of CO and NOx concentrations observed in the beginning of the '90s is almost entirely attributed to the increase of the catalyst-equipped cars during this period. The numerical parameters of an empirical model relating EU standard exceedances with mean annual concentrations were defined and the model was validated using datasets from years that were not used for the estimation of these parameters. This model was used in conjunction with a roll-back equation as a policy tool for the assessment of the effect of different CO and NOx emissions reduction scenarios on air quality standard compliance for CO and NO2. Results predicted with this empirical modelling approach were assessed with monitored data averaged over a 3-year period, giving satisfactory results. Conclusion A methodology suggested for assessing the effects of different emissions reduction scenarios on air quality standard attainment was successfully applied for CO and NO2 in the Athens area. Recommendation and Perspective The proposed methodology can provide a useful tool for the evaluation of policies already in progress as well as the development of future policies for emissions reduction in urban areas with similar characteristics, aiming at air quality standard compliance on a timely manner. Such a methodology could be applied in other urban areas of Greece characterized by dense traffic, therefore assisting the development of national policies in relation to air pollutants for which standard exceedances occur.

GOALS, SCOPE AND BACKGROUND: It has been observed that hydrocarbon treated wastewaters still contain high COD and a number of intermediates. This suggests that the required catabolic gene pool for further degradation might be absent in the system or, that its titer value is not significant enough. By providing the desired catabolic potential, the overall efficiency of the treatment system can be improved. This study aims to demonstrate this concept by bioaugmentation of a lab-scale reactor treating refinery wastewater with a consortium having the capacity to complement the alkB genotype to the available microbial population. METHODS: Two reactors were set up using activated biomass collected from a refinery treatment plant and operated at a continuous mode for a period of 8 weeks. The feed to both reactors was kept constant. Crude oil was spiked regularly. One reactor was bioaugmented with a consortium previously described for crude oil spill remediation. The efficiency of the bioaugmented reactor was demonstrated by reduced COD. The changes in the microbial population over a period of time were analyzed by RAPD. Catabolic activity of the biomass in both reactors was monitored by PCR. The presence of the catabolic loci was confirmed by Southern Hybridization. RESULTS AND DISCUSSION: 52.2% removal of COD was observed in the bioaugmented reactor while only 15.1% reduction of COD was observed in the reactor without bioaugmentation. The change in microbial population can be seen from the 4th week, which also corresponds to improved catabolic activity. The presence of the bedA locus was seen in all samples, which indicates the presence of aromatic degraders, but the appearance of the alkB locus, from the 6th week onwards, which was observed only in the samples from the bioaugmented reactor. The results suggest that the gene pool of the bioaugmented reactor has catabolic loci that can degrade accumulated intermediates, thus improving the efficiency of the system. CONCLUSIONS: In this study, improvement of efficiency of bioremediation was demonstrated by addition of catabolic loci that are responsible for degradation. Bioaugmentation was carried out in biomass that was collected from an ETP (effluent treatment plant) treating hydrocarbon containing wastewater to study the strategies for improvement of the treatment system. Biostimulation, only marginally improved the efficiency, when compared to bioaugmentation. The improved efficiency was demonstrated by COD removal. The presence of the alkB locus suggests the importance of a catabolic gene pool that acts on accumulated intermediates. It is well documented that straight chain aliphatics and intermediates of aromatic compounds after ring cleavage, accumulate in refinery wastewater systems, thereby hindering further degradation of the wastewater. Supplementation of a catabolic gene pool that treats the lower pathway compounds and alkanes will improve the overall efficiency. In this study, results suggest that the alkB locus can also be used to monitor the degradative mode of the activated biomass. RECOMMENDATIONS AND PERSPECTIVE: . Pollution from petroleum and petroleum products around the globe are known to have grave consequences on the environment. Bioremediation, using activated sludge, is one option for the treatment of such wastes. Effluent treatment plants are usually unable to completely degrade the wastewater being treated in the biological unit (the aerator chambers). The efficiency of degradation can be improved by biostimulation and bioaugmentation. This study demonstrates the improved efficiency of a treatment system for wastewater containing hydrocarbons by bioaugmentation of a consortium that supports degradation. Further experiments on a pilot scale are recommended to assess the use of bioaugmentation on a large scale. The use of molecular tools, like DNA probes for alkB, to monitor the system also needs to be explored.

BACKGROUND AND AIM: In non-eutrophicated freshwaters, humic substances (HS) pose chemical stresses on aquatic organisms and, hence, separating sensitive from less sensitive or even tolerant species. One of the stresses, identified so far, is the reduction of photosynthetic oxygen production and reduction in growth in freshwater macrophytes and algae. In a previous paper, it has been shown that even closely related coccal green algae responded differently upon identical stress by HS, which is consistent with the hypothesis above. Due to their much simpler cellular ultrastructure, cyanobacteria are supposed to be more sensitive to HS exposure than eukaryote should be. One coccal green algal species (Desmodesmus communis) and one cyanobacterium (Chroococcus minutus) were exposed to four natural organic matter (NOM) isolates. One NOM has been isolated from a brown water lake (Schwarzer See) in Brandenburg State; three were obtained from a comprehensive Scandinavian NOM research project and originated from Norway (Birkenes), Finland (Hietajärvi), and Sweden (Svartberget). METHODS: Cultures of D. communis and C. minutus were obtained from the Culture Collection of Algae, Göttingen, and maintained in a common medium. The cultures were non-axenic. The algae and cyanobacteria were exposed under identical conditions to environmentally realistic NOM concentrations. Cell numbers were counted microscopically in Neugebauer cuvettes in triplicates. To avoid limitation by nutrient depletion, the experiments were terminated after 14 to 15 days. Until culture day 12, no growth limitation has been observed in the controls. RESULTS: All NOM isolates modulated the growth of the algae and cyanobacteria. During the early culture days, there was a slight growth promotion with the coccal green alga and to a much lesser degree with the cyanobacterium. Yet, the major effect were significant reductions in cell yield in both primary producer cultures. C. minutus was much more affected than D. communis. This applies particularly to the three tested Scandinavian NOM isolates, which were effective at concentrations even below 1 mg L-1 DOC. DISCUSSION: The growth promoting effect may be due to an increase in bioavailability of some trace nutrients in the presence of NOM, the release of some growth promoting substances by microbial or photochemical processing of the NOM, and/or a hormetic effect. The growth reducing effect can be explained as a herbicide-like mode of action that affects the photosystem II most prevalent by blocking the electron transport chain, absorption of electrons, or production of an internal oxidative stress after processing the bioconcentrated HS. Furthermore, it may be postulated that also phototoxicity of these HS in the algal cells contributes to the overall toxicity; however, experimental evidence is lacking so far. CONCLUSIONS: Upon exposure to HS, cyanobacteria appear to be much more sensitive than coccal green algae and respond in growth reduction. This high sensitivity of cyanobacteria to HS may explain phytoplankton patterns in the field. Eutrophic, humic-rich lakes do not support the cyanobacterial blooms characteristic of eutrophic, but humic-poor lakes. In the humic-rich systems, raphidophytes or, less frequent, specific coccal greens are more common. Obviously, cyanobacteria appear to be unable to make advantage of their accessory pigments (phycocyanin) to exploit the reddish light prevailing in humic-rich lakes. RECOMMENDATION: At present, no effective structure can be figured out which may be responsible for the adverse effect on the cyanobacterial species. It is reserved to future research whether or not HS may be applied more specifically (for instance, with elevated moieties of the effective structures) as a natural geochemical to combat cyanobacterial blooms.

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.

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 SeO ₄ ²⁻ and from 0.50 to 10.0 mg Se/L for the treatments exposed to SeO ₃ ²⁻ , 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 SeO ₃ ²⁻ 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 SeO ₄ ²⁻ and SeO ₃ ²⁻ , 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; SeO ₃ ²⁻ caused much more severe stress to plants than SeO ₄ ²⁻ 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 SeO ₄ ²⁻ and SeO ₃ ²⁻ 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 SeO ₄ ²⁻ than for SeO ₃ ²⁻ . 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.

BACKGROUND, AIM AND SCOPE: Discussions on management of whales and whaling are factually monopolized by the International Whaling Commission (IWC), resulting in a limitation of information flow to outside communities. With an aim to improve the situation, this article briefly reviews whaling and dolphin/porpoise fisheries in Japan, which is recognized to be the world largest cetacean exploitation.MAIN FEATURES: The Japanese government grants an annual take of 22,647 cetaceans of 15 species for scientific whaling and various kinds of active dolphin/porpoise fisheries by the nationals. Further, over 100 baleen whales and numerous small cetaceans are taken in passive net fisheries. They are used mostly for human consumption and some for aquarium display.Results. Sustainability of the take is not evident and some populations have shown a historical decline. The Japanese program of scientific whaling has been reviewed by IWC and its Scientific Committee (SC), although they have arrived at no consensus. DISCUSSION: The current scientific whaling program invites arguments from the view points of science as well as concerning the ethics of scientists, economy, and interpretation of the International Convention for Regulation of Whaling (ICRW) of 1946. The scientific whaling and other Japanese cetacean fisheries are benefited from nationalistic public attitude, and ambiguity and weakness of the ICRW. CONCLUSIONS: Japanese cetacean harvest will continue supported by domestic demand for whale products as long as the proceeds can sustain the operation, even with criticisms from outside communities. RECOMMENDATIONS AND PERSPECTIVE: . For safe management of small cetaceans exploited by Japan, studies are urgent on the population structure, abundance and validity of catch statistics. The results should be open to scientific communities.