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

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.

BACKGROUND, AIM AND SCOPE: The presence of heavy metals in wastewater is one of the main causes of water and soil pollution. The aim of the present study was to investigate the removal of Cd, Cu, Pb, Hg, Mn, Cr and Zn in urban effluent by a biological wastewater treatment, as well as to quantify the levels of As, Be, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Sn, Tl, V and Zn in dewatering sludge from the Biological Wastewater Treatment Plant to Ribeirão Preto (RP-BWTP), Brazil. MATERIALS AND METHODS: Concentrations of Cd, Cr, Cu, Mn and Pb in wastewater and those of Ni in sludge were determined by atomic absorption spectrophotometry with graphite furnace atomization. Mercury concentrations in wastewater were measured by hydride generation atomic spectrophotometry, and Zn levels were determined by atomic absorption spectrophotometry using acetylene flame. In sludge, the levels of As, Be, Cd, Cr, Cu, Fe, Hg, Mn, Pb, Sn, Tl, V and Zn were determined by inductively coupled plasma-mass spectrometry. RESULTS: The percentages of removal efficiency (RE) were the following: Hg 61.5%, Cd 60.0%, Zn 44.9%, Cu 44.2%, PB 39.7%, Cr 16,5% and Mn 10.4%. In turn, the mean concentrations (mg/kg) of metals in dewatering sludge followed this increasing order: Tl (<0.03), Hg (0.31), Be (0.43), As (1.14), Cd (1.34), V (59.2), Pb (132.1), Sn (166.1), Cr (195.0), Mn (208.1), Ni (239.4), Cu (391.7), Zn (864.4) and Fe (20537). DISCUSSION: The relationship between metal levels in untreated wastewater, as well as the removal efficiency are in agreement with previous data from various investigators, It is important to note that metal removal efficiency is not only affected by metal ion species and concentration, but also by other conditions such as operating parameters, physical, chemical, and biological factors. CONCLUSIONS: Metal values recorded for treated wastewater and sludge were within the maximum permitted levels established by the Environmental Sanitation Company (CETESB), São Paulo, Brazil. RECOMMENDATIONS: There is an urgent need for the authorities who are responsible for legislation on sludge uses in agriculture of establishing safety levels for As, Be, Hg, Sn, Tl and V. PERSPECTIVES: According to the current metal levels, RP-BWTP sludge might be used for agriculture purposes. However, for an environmentally safe use of sewage sludge, further studies including systematic monitoring are recommended. Annual metal concentrations and predicted variations of those elements in the sludge should be monitored.

BACKGROUND: Goals and Scope. In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the 'Special Activity on the Testing and Assessment of Endocrine Disruptors' within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. -: Study Design. Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. RESULTS: All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-response manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC50s calculated were comparable (coefficients of variation 34-49%) for all hormones. DISCUSSION: and Perspectives. The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible across all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 pre-validation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol across the same group of laboratories using an extended set of chemicals (~30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways.

GOAL, SCOPE AND BACKGROUND: Sewage sludge produced in wastewater treatment contains large amounts of organic matter and nutrients and could, therefore, be suitable as fertiliser. However, with the sludge, besides heavy metals and pathogenic bacteria, a variety of organic contaminants can be added to agricultural fields. Whether the organic contaminants from the sludge can have adverse effects on human health and wildlife if these compounds enter the food chain or groundwater still remains a point of controversial discussion. MAIN FEATURES: This paper presents an overview on the present situation in Europe and a summary of some recent results on the possible uptake of organic contaminants by crops after addition to agricultural fields by sewage sludge. RESULTS: Greenhouse experiments and field trials were performed to study the degradation and uptake of organic micro-contaminants in sludge-amended agricultural soil in crops, such as barley and carrots grown in agricultural soil amended with anaerobically-treated sewage sludge from a wastewater treatment plant, but studies hitherto have revealed no immediate risks. Common sludge contaminants such as linear alkylbenzene sulphonates (LAS), nonylphenol ethoxylates (NPE), polycyclic aromatic hydrocarbons (PAH), bis(diethylhexyl) phthalate (DEHP), showed neither accumulation in soil nor uptake in plants. DISCUSSION: It is assumed that the annual amount of sewage sludge produced in Europe will increase in the future, mainly due to larger amounts of high quality drinking water needed by an increasing population and due to increasing demands for cleaner sewage water. Application of sewage sludge to agricultural soils is sustainable and economical due to nutrient cycling and disposal of sewage sludge. However, this solution also involves risks with respect to the occurrence of organic contaminants and other potentially harmful contents such as pathogens and heavy metals present in the sludge. There have been concerns that organic contaminants may accumulate in the soil, be taken up by plants and thereby transferred to humans via the food chain. Results obtained so far revealed, however, no immediate risk of accumulation of common organic sludge contaminants in soil or uptake in plants when applying sewage sludge to agricultural soil. With very high dosages of sewage sludge, there may be a risk for accumulation of very apolar contaminants, such as DEHP, to the soil. CONCLUSIONS: Any conclusions on the safe use of sewage sludge in agriculture have to be drawn carefully, as the studies performed until now have been limited. Further studies are required, and before final statements can be drawn, it is imminent to study a larger variety of common crops and the effect sewage sludge application may have on a possible accumulation of organic contaminants in the crops. Furthermore, a larger variety of organic contaminants need to be studied and special focus should be given to contaminants newly introduced into the environment. Besides investigating possible plant uptake of organic contaminants, the fate of these compounds in soil after sludge application need to be monitored too. Here, special attention has to be given to studies on degradation and the formation of degradation products, to weathering and to leaching effects on groundwater, to the application of different crops on the same field (crop rotation), to the use of full-width tillage and strip tillage, and to long term application of sewage sludge on the soil. RECOMMENDATIONS AND PERSPECTIVE: There are environmental, political as well as economical incentives to increase the agricultural application of sludge. However, such usage should be performed with care as there are also ways in which sludge fertilisation could harm the environment and human health. Recently, a new European COST Action (859) has been established covering the field of food safety and improved food quality. Part of the Action is dealing with the application of sewage sludge in agriculture. Before any political and economical measures can be taken, the pros and cons have to be sufficiently investigated on a scientific level first.

BACKGROUND AND AIM: An accurate estimation of biogenic emissions of VOC (volatile organic compounds) is necessary for better understanding a series of current environmental problems such as summertime smog and global climate change. However, very limited studies have been reported on such emissions in China. The aim of this paper is to present an estimate of biogenic VOC emissions during summertime in China, and discuss its uncertainties and potential areas for further investigations. MATERIALS AND METHODS: This study was mainly based on field data and related research available so far in China and abroad, including distributions of land use and vegetations, biomass densities and emission potentials. VOC were grouped into isoprene, monoterpenes and other VOC (OVOC). Emission potentials of forests were determined for 22 genera or species, and then assigned to 33 forest ecosystems. The NCEP/NCAR reanalysis database was used as standard environmental conditions. A typical summertime of July 1999 was chosen for detailed calculations. RESULTS AND DISCUSSION: The biogenic VOC emissions in China in July were estimated to be 2.3×1012gC, with 42% as isoprene, 19% as monoterpenes and 39% as OVOC. About 77.3% of the emissions are generated from forests and woodlands. The averaged emission intensity was 4.11 mgC m-2 hr-1 for forests and 1.12 mgC m-2 hr-1 for all types of vegetations in China during the summertime. The uncertainty in the results arose from both the data and the assumptions used in the extrapolations. Generally, uncertainty in the field measurements is relatively small. A large part of the uncertainty mainly comes from the taxonomic method to assign emission potentials to unmeasured species, while the ARGR method serves to estimate leaf biomass and the emission algorithms to describe light and temperature dependence. CONCLUSIONS: This study describes a picture of the biogenic VOC emissions during summertime in China. Due to the uneven spatial and temporal distributions, biogenic VOC emissions may play an important role in the tropospheric chemistry during summertime. RECOMMENDATIONS AND PERSPECTIVE: Further investigations are needed to reduce uncertainties involved in the related factors such as emission potentials, leaf biomass, species distribution as well as the mechanisms of the emission activities. Besides ground measurements, attention should also be placed on other techniques such as remote-sensing and dynamic modeling. These new approaches, combined with ground measurements as basic database for calibration and evaluation, can hopefully provide more comprehensive information in the research of this field.

BACKGROUND, AIM AND SCOPE: Agricultural industrial wastes can frequently be used as raw materials in the production of bio-fuels. Olive-oil milling wastewater is considered as one of the most polluting agro-industrial residues, but fortunately due to its high content of organic matter, it has the potential to be a valuable starting material to obtain bio-ethanol via fermentation with Saccharomyces cerevisiae. The fermentation of olive-oil milling wastewater has been demonstrated with good yield of ethanol (8-12%v/v) once the level of reducing sugars is appropriate and the phenolic fraction, which inhibits the yeast, is removed.Materials and Methods:-Results:-Discussion:-Conclusions:-Recommendations and Perspectives:-