Physiological and biochemical responses of <Emphasis Type="Italic">Machilus ichangensis</Emphasis> Rehd. et Wils and <Emphasis Type="Italic">Taxus chinensis</Emphasis> (Pilger) Rehd. to elevated O<Subscript>3</Subscript> in subtropical China

Considerable researches have documented the negative effects of ozone on woody species in North America and Europe; however, little is known about how woody tree species respond to elevated O₃ in subtropical China, and most of the previous studies were conducted using pot experiment. In the present study, Machilus ichangensis Rehd. et Wils (M. ichangensis) and Taxus chinensis (Pilger) Rehd. (T. chinensis), evergreen tree species in subtropical China, were exposed to non-filtered air (NF), 100 nmol mol^?1 O₃ (E1) and 150 nmol mol^?1 O₃ (E2), in open-top chambers under field conditions from 21st March to 2nd November 2015. In this study, O₃ fumigation significantly reduced net photosynthesis rate (Pn) in M. ichangensis in the three measurements and in T. chinensis in the last measurement. Also, non-stomatal factors should be primarily responsible for the decreased Pn. O₃ fumigation-induced increase in malondialdehyde, superoxide dismutase, and reduced ascorbic acid levels indicated that antioxidant defense mechanism had been stimulated to prevent O₃ stress and repair the oxidative damage. Yet, the increase of antioxidant ability was not enough to counteract the harm of O₃ fumigation. Because of the decrease in CO₂ assimilation, the growth of the two tree species was restrained ultimately. The sensitivity of the two tree species to O₃ can be determined: M. ichangensis > T. chinensis. It suggests a close link between the rising O₃ concentrations and the health risk of some tree species in subtropics in the near future.     

Zinc tolerance and uptake by <Emphasis Type="Italic">Arabidopsis halleri ssp. gemmifera</Emphasis> grown in nutrient solution

Background, aim, and scope  

Zinc is an essential micronutrient element but its concentrations found in contaminated soils frequently exceed those required by the plant and soil organisms, and thus create danger to animal and human health. Phytoremediation is a technique, often employed in remediation of contaminated soils, which aims to remove heavy metals or other contaminants from soils or waters using plants. Arabidopsis (A.) halleri ssp. gemmifera is a plant recently found to be grown vigorously in heavy metal contaminated areas of Japan and it contained remarkably high amount of heavy metals in its shoots. However, the magnitude of Zn accumulation and tolerance in A. halleri ssp. gemmifera need to be investigated for its use as a phytoremediation plant.     

Interaction of As and Sb in the hyperaccumulator <Emphasis Type="Italic">Pteris vittata</Emphasis> L.: changes in As and Sb speciation by XANES

Arsenic (As) and antimony (Sb) are chemical analogs that display similar characteristics in the environment. The As hyperaccumulator Pteris vittata L. is a potential As–Sb co-accumulating species. However, when this plant is exposed to different As and Sb speciation, the associated accumulating mechanisms and subsequent assimilation processes of As and Sb remain unclear. A 2-week hydroponic experiment was conducted by exposing P. vittata to single AsIII, AsV, SbIII, and SbV or the co-existence of AsIII and SbIII and AsV and SbV. P. vittata could co-accumulate As and Sb in the pinna (>1000 mg kg^?1) with high translocation (>1) of As and Sb from the root to the pinna. P. vittata displayed apparent preference to the trivalent speciation of As and Sb than to the pentavalent speciation. Under the single exposure of AsIII or SbIII, the pinna concentration of As and Sb was 84 and 765 % higher than that under the single exposure of AsV or SbV, respectively. Despite the provided As speciation, the main speciation of As in the root was AsV, whereas the main speciation of As in the pinna was AsIII. The Sb in the roots comprised SbV and SbIII when exposed to SbV but was exclusively SbIII when exposed to SbIII. The Sb in the pinna was a mixture of SbV and SbIII regardless of the provided Sb speciation. Compared with the single exposure of As, the co-existence of As and Sb increased the As concentration in the pinna of P. vittata by 50–66 %, accompanied by a significant increase in the AsIII percentage in the root. Compared with the single exposure of Sb, the co-existence of Sb and As also increased the Sb concentration in the pinna by 51–100 %, but no significant change in Sb speciation was found in P. vittata.     

Effects of acetaminophen in <Emphasis Type="Italic">Brassica juncea</Emphasis> L. Czern.: investigation of uptake,translocation, detoxification,and the induced defense pathways

Purpose  

Besides classical organic pollutants and pesticides, pharmaceuticals and their residues have nowadays become recognized as relevant environmental contaminants. The risks of these chemicals for aquatic ecosystems are well known, but information about the pharmaca-plant interactions and metabolic pathways is scarce. Therefore, we investigate the process of uptake of acetaminophen (N-Acetyl-4-aminophenol) by Brassica juncea, drug-induced defense responses and detoxification mechanisms in different plant parts.     

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.     

Investigation of low-level <Superscript>242</Superscript>Pu contamination on nutrition disturbance and oxidative stress in <Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.

Plutonium associated with higher molecular weight molecules is presumed to be poorly mobile and hardly plant available. In our present study, we investigate the uptake and effects of Pu treatments on Solanum tuberosum plants in amended Hoagland medium at concentrations of [²⁴²Pu] = 100 and 500 nm, respectively. We found a direct proof of oxidative stress in the plants caused by these rather low concentrations. For the confirmation of oxidative stress, we explored the production of nitric oxide (NO) and hydrogen peroxide (H₂O₂) by epifluorescence microscopy. Oxidative stress markers like lipid peroxidation and superoxide radicals (O₂ ^??) are monitored through histochemical analysis. The biochemical parameters i.e. chlorophyll and carotenoids are measured as an indicator of cellular damage in the tested plants including the enzymatic parameters such as catalase and glutathione reductase. From our work, we conclude that Pu in low concentration has no significant effects on the uptake of many trace and macroelements. In contrast, the content of O₂ ^?? , malondialdehyde (MDA), and H₂O₂ increases with increasing Pu concentration in the solution, while the opposite effects was found for NO, catalase, and glutathione reductase. These findings prove that even low concentration of Pu regulates ROS production and generate oxidative stress in S. tuberosum L.     

Metal uptake capability of <Emphasis Type="Italic">Cyperus articulatus</Emphasis> L. and its role in mitigating heavy metals from contaminated wetlands

Wetland plants are biological filters that play an important role in maintaining aquatic ecosystem and can take up toxic metals from sediments and water. The present study investigated the seasonal variation in the accumulation potential of heavy metals by Cyperus articulatus in contaminated watercourses. Forty quadrats, distributed equally in 8 sites (six contaminated sites along Ismailia canal and two uncontaminated sites along the River Nile), were selected seasonally for sediment, water, and plant investigations. Autumn was the flourishing season of C. articulatus with the highest shoot density, length, and diameter as well as aboveground biomass, while summer showed the least growth performance. The photosynthetic pigments were markedly reduced under contamination stress. C. articulatus plants accumulated concentrations of most heavy metals, except Pb, in their roots higher than the shoots. The plant tissues accumulated the highest concentrations of Fe, Cd, Ni, and Zn during autumn, while Cu and Mn during spring, and Cr and Co during winter. It was found that Cd, Cu, Ni, Zn, Pb, and Co had seasonal bioaccumulation factor (BF) > 1 with the highest BF for Cd, Ni, and Zn during autumn, Co, Cu, and Pb in winter, spring, and summer, respectively. The translocation factor of most heavy metals, except Pb in spring, was <1 indicating potential phytostabilization of these metals. In conclusion, autumn is an ideal season for harvesting C. articulatus in order to monitor pollution in contaminated wetlands.     

Uranium biosorption by <Emphasis Type="Italic">Padina</Emphasis> sp. algae biomass: kinetics and thermodynamics

Introduction  

Kinetic, thermodynamic, and equilibrium isotherms of the biosorption of uranium ions onto Padina sp., a brown algae biomass, in a batch system have been studied.     

Biomonitoring the genotoxic effects of pollutants on <Emphasis Type="Italic">Tradescantia pallida</Emphasis> (Rose) D.R. Hunt in Dourados,Brazil

Purpose  

This study aimed to associate the intensity of vehicular traffic in the city of Dourados (Mato Grosso do Sul State, Brazil) with mutagenic effects and alterations in leaf physiology as measured by the quantity of micronuclei and the leaf surface parameters of Tradescantia pallida.     

Green synthesis of gold nanoparticles using fungus <Emphasis Type="Italic">Mariannaea</Emphasis> sp. HJ and their catalysis in reduction of 4-nitrophenol

In the present study, biosynthesis of gold nanoparticles (AuNPs) by the cells (cells-AuNPs) and cell-free extracts (extracts-AuNPs) of a new fungus Mariannaea sp. HJ was reported. The as-synthesized particles were characterized by UV-vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The effects of different parameters on AuNP biosynthesis were investigated, and initial gold ion concentration of 2 mM, pH 7, was demonstrated to be suitable for both cells-AuNP and extracts-AuNP syntheses. The cells-AuNPs were of various shapes, including sphere, hexagon, and irregular shapes, with an average size of 37.4 nm, while the extracts-AuNPs were almost spherical and pseudo-spherical with an average size of 11.7 nm. XRD pattern suggested that the crystal structure of both AuNPs was face-centered cubic. FTIR spectra implied that some biomolecules from the fungal cell walls or cell-free extracts were involved in the formation of AuNPs. The as-synthesized AuNPs were demonstrated to have excellent catalytic activities for the reduction of 4-nitrophenol with the catalytic rate constants of 5.7 × 10^?3/s for cells-AuNPs and 24.7 × 10^?3/s for extracts-AuNPs. To the best of our knowledge, this is the first report on AuNP biosynthesis by Mariannaea sp.     

Behaviour and dynamics of di-ammonium phosphate in bauxite processing residue sand in Western Australia—I. NH<Subscript>3</Subscript> volatilisation and residual nitrogen availability

Background, aim and scope

Australia is the largest producer of bauxite in the world, with an annual output of approximately 62 million metric dry tons in 2007. For every tonne of alumina, about 2 tonnes of highly alkaline and highly saline bauxite-processing residue are produced. In Western Australia, Alcoa World Alumina, Australia (Alcoa) produces approximately 15 MT of residue annually from its refineries (Kwinana, Pinjarra and Wagerup). The bauxite-processing residue sand (BRS) fraction represents the primary material for rehabilitating Alcoa’s residue disposal areas (RDAs). However, the inherently hostile characteristics (high alkalinity, high salinity and poor nutrient availability) of BRS pose severe limitations for establishing sustainable plant cover systems. Alcoa currently applies 2.7 t ha^?1 of di-ammonium phosphate ((NH₄)₂HPO₄; DAP)-based fertiliser as a part of rehabilitation of the outer residue sand embankments of its RDAs. Limited information on the behaviour of the dominant components of this inorganic fertiliser in highly alkaline BRS is currently available, despite the known effects of pH on ammonium (NH₄) and phosphorus (P) behaviour. The aim of this study was to quantify the effects of pH on NH₃ volatilisation and residual nitrogen (N) in BRS following DAP applications.

Methods

The sponge-trapping and KCl-extraction method was used for determining NH₃ volatilisation from surface-applied DAP in samples of BRS collected from each of Alcoa’s three Western Australia Refineries (Kwinana, Pinjarra, Wagerup) under various pH conditions (pH 4, 7, 9 and 11). Following cessation of volatilisation, the residual N was extracted from BRS using 2 M KCl and concentrations of NH ₄ ⁺ –N and NO ₃ ^? –N were determined by flow injection analysis.

Results

The quantities of NH₃ volatilised increased dramatically as the pH increased from 4 to 11. Much of the N lost as NH₃ (up to 95.2%) occurred within a short period (24 h to 7 days), particularly for the pH 9 and 11 treatments. Concentrations of residual NH ₄ ⁺ –N recovered in DAP-treated BRS at the end of the experiment decreased with increasing pH. This finding was consistent with increasing loss of N via volatilisation as pH increased. The concentration of NO ₃ ^? –N was very low due to no nitrification in BRS.

Discussion

The pH was a key driver for NH₃ volatilisation from DAP-treated BRS and primarily controlled N dynamics in BRS. Results indicate that NH₄ not adsorbed by BRS was highly susceptible to volatilisation. The likely lack of nitrifying bacteria did not allow conversion of ammonium to nitrate, thereby further exacerbating the potential for loss via volatilisation

Conclusions

It was demonstrated that the pH is the key factor controlling the loss of inorganic N from BRS. Although volatilisation was considerably lower at pH 4, achieving this pH reduction in the field is not possible at present. Findings from this study highlight the need to better understand which forms of N fertiliser are most suitable for use in highly alkaline BRS.

Recommendation and perspectives

Although pH reduction is the most likely means of stopping NH₃ volatilisation in BRS, it is economically and operationally unfeasible to add sufficient acidity for adequately lowering pH in the BRS for revegetation. More attention on forms of fertilisers more suitable to highly alkaline, microbially inert soil conditions appears to be warranted.

     

Impact of metal stress on the production of secondary metabolites in <Emphasis Type="Italic">Pteris vittata</Emphasis> L. and associated rhizosphere bacterial communities

Plants adapt to metal stress by modifying their metabolism including the production of secondary metabolites in plant tissues. Such changes may impact the diversity and functions of plant associated microbial communities. Our study aimed to evaluate the influence of metals on the secondary metabolism of plants and the indirect impact on rhizosphere bacterial communities. We then compared the secondary metabolites of the hyperaccumulator Pteris vittata L. collected from a contaminated mining site to a non-contaminated site in Vietnam and identified the discriminant metabolites. Our data showed a significant increase in chlorogenic acid derivatives and A-type procyanidin in plant roots at the contaminated site. We hypothesized that the intensive production of these compounds could be part of the antioxidant defense mechanism in response to metals. In parallel, the structure and diversity of bulk soil and rhizosphere communities was studied using high-throughput sequencing. The results showed strong differences in bacterial composition, characterized by the dominance of Proteobacteria and Nitrospira in the contaminated bulk soil, and the enrichment of some potential human pathogens, i.e., Acinetobacter, Mycobacterium, and Cupriavidus in P. vittata’s rhizosphere at the mining site. Overall, metal pollution modified the production of P. vittata secondary metabolites and altered the diversity and structure of bacterial communities. Further investigations are needed to understand whether the plant recruits specific bacteria to adapt to metal stress.     

Biodegradation of C.I. Acid Red 1 by indigenous bacteria <Emphasis Type="Italic">Stenotrophomonas</Emphasis> sp. BHUSSp X2 isolated from dye contaminated soil

A significant proportion of xenobiotic recalcitrant azo dyes are being released in environment during carpet dyeing. The bacterial strain Stenotrophomonas sp. BHUSSp X2 was isolated from dye contaminated soil of carpet industry, Bhadohi, India. The isolated bacterial strain was identified morphologically, biochemically, and on the basis of 16S rRNA gene sequence. The isolate decolorized 97 % of C.I. Acid Red 1 (Acid RED G) at the concentration of 200 mg/l within 6 h under optimum static conditions (temperature ?35 °C, pH 8, and initial cell concentration 7?×?10⁷ cell/ml). Drastic reduction in dye degradation rate was observed beyond initial dye concentration from 500 mg/l (90 %), and it reaches to 25 % at 1000 mg/l under same set of conditions. The analysis related to decolorization and degradation was done using UV-Vis spectrophotometer, HPLC, and FTIR, whereas the GC-MS technique was utilized for the identification of degradation products. Phytotoxicity analysis revealed that degradation products are less toxic as compared to the original dye.     

Isolation and characterization of <Emphasis Type="Italic">Bradyrhizobium</Emphasis> sp. SR1 degrading two β-triketone herbicides

In this study, a bacterial strain able to use sulcotrione, a β-triketone herbicide, as sole source of carbon and energy was isolated from soil samples previously treated with this herbicide. Phylogenetic study based on16S rRNA gene sequence showed that the isolate has 100 % of similarity with several Bradyrhizobium and was accordingly designated as Bradyrhizobium sp. SR1. Plasmid profiling revealed the presence of a large plasmid (>50 kb) in SR1 not cured under nonselective conditions. Its transfer to Escherichia coli by electroporation failed to induce β-triketone degrading capacity, suggesting that degrading genes possibly located on this plasmid cannot be expressed in E. coli or that they are not plasmid borne. The evaluation of the SR1 ability to degrade various synthetic (mesotrione and tembotrione) and natural (leptospermone) triketones showed that this strain was also able to degrade mesotrione. Although SR1 was able to entirely dissipate both herbicides, degradation rate of sulcotrione was ten times higher than that of mesotrione, showing a greater affinity of degrading-enzyme system to sulcotrione. Degradation pathway of sulcotrione involved the formation of 2-chloro-4-mesylbenzoic acid (CMBA), previously identified in sulcotrione degradation, and of a new metabolite identified as hydroxy-sulcotrione. Mesotrione degradation pathway leads to the accumulation of 4-methylsulfonyl-2-nitrobenzoic acid (MNBA) and 2-amino-4 methylsulfonylbenzoic acid (AMBA), two well-known metabolites of this herbicide. Along with the dissipation of β-triketones, one could observe the decrease in 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibition, indicating that toxicity was due to parent molecules, and not to the formed metabolites. This is the first report of the isolation of bacterial strain able to transform two β-triketones.     

Statistical analysis and isotherm study of uranium biosorption by <Emphasis Type="Italic">Padina</Emphasis> sp. algae biomass

Introduction  

The application of response surface methodology is presented for optimizing the removal of U ions from aqueous solutions using Padina sp., a brown marine algal biomass.     

Biomass reduction and arsenic transformation during composting of arsenic-rich hyperaccumulator <Emphasis Type="Italic">Pteris vittata</Emphasis> L.

Background, aim, and scope  

Composting is being proposed as a pretreatment step before disposal of metal-rich biomass after phytoextraction process. This study determined the biomass reduction and arsenic transformation during composting As-rich biomass of hyperaccumulator Chinese brake fern (Pteris vittata L.).     

Combined bioremediation of atrazine-contaminated soil by <Emphasis Type="Italic">Pennisetum</Emphasis> and <Emphasis Type="Italic">Arthrobacter</Emphasis> sp. strain DNS10

Strain DNS10 was isolated from the black soil collected from the northeast of China which had been cultivated with atrazine as the sole nitrogen source. Pennisetum is a common plant in Heilongjiang Province of China. The main objective of this paper was to evaluate the efficiency of plant–microbe joint interactions (Arthrobacter sp. DNS10 + Pennisetum) in atrazine degradation compared with single-strain and single-plant effects. Plant–microbe joint interactions degraded 98.10 % of the atrazine, while single strain and single plant only degraded 87.38 and 66.71 % after a 30-day experimental period, respectively. The results indicated that plant–microbe joint interactions had a better degradation effect. Meanwhile, we found that plant–microbe joint interactions showed a higher microbial diversity. The results of microbial diversity illustrated that the positive effects of cropping could improve soil microbial growth and activity. In addition, we planted atrazine-sensitive plants (soybean) in the soil after repair. The results showed that soybean growth in soil previously treated with the plant–microbe joint interactions treatment was better compared with other treatments after 20 days of growth. This was further proved that the soil is more conducive for crop cultivation. Hence, plant–microbe joint interactions are considered to be a potential tool in the remediation of atrazine-contaminated soil.     

Occurrence and temporal variations of the xenoestrogens bisphenol A, 4-<Emphasis Type="Italic">tert</Emphasis>-octylphenol,and tech. 4-nonylphenol in two German wastewater treatment plants

GOAL, SCOPE, AND BACKGROUND: The xenoestrogens bisphenol A, 4-tert-octylphenol, and the technical isomer mixture of 4-nonylphenol (tech. 4-nonylphenol) belong to the group of chemicals which are called endocrine disrupters due to their property of causing hormonal dysfunctions in the endocrine system of organisms at very low concentrations. Bisphenol A, 4-tert-octylphenol, and the tech. 4-nonylphenol (mixture of isomers) were determined in water samples collected from the influent and effluent of two German wastewater treatment plants (WWTP) during a long-time sampling period from February 2003 till August 2005 to assess their occurrence and temporal variations in WWTPs. METHODS: The compounds were extracted and concentrated from water by solid-phase extraction (SPE) using Bond Elut PPL cartridges and quantified by use of gas chromatography-mass spectrometry (GC-MS). RESULTS: The influent concentrations were as follows: Bisphenol A < limit of detection of the method (< ldm)--12,205 ng L(-1), tech. 4-nonylphenol < ldm--10,186 ng L(-1), and 4-tert-octylphenol 39-1,495 ng L(-1). The measured effluent concentrations were lower with values in the range of < ldm--7,625 ng L(-1) for bisphenol A, < ldm--14,444 ng L(-1) for tech. 4-nonylphenol, and < ldm--392 ng L(-1) for 4-tert-octylphenol. All target compounds were largely eliminated during the wastewater treatment process. The elimination efficiency varied between 73% and 93%. DISCUSSION: All analytes show highly fluctuating influent concentrations with very high peak concentrations at particular sampling times. The variation of effluent concentrations is by far lower than the variation of influent concentrations. For tech. 4-nonylphenol, a significant temporal concentration variation has been detected with very high concentrations up to the microgram-per-liter level in the time from February 2003 till July 2003 and clearly decreasing concentrations in the time from June 2004 till August 2005. This corresponds well with the implementation of Directive 2003/53/EC (nonylphenol and nonylphenol ethoxylates in the European Union "may not be placed on the marked or used as a substance or constituent of preparations in concentrations equal or higher than 0.1% by mass") from January 2005 on. Bisphenol A is present in the effluent samples in a wide range of concentrations from below the detection limit to high concentrations up to the microgram-per-liter level. For 4-tert-octylphenol, no particular trend of concentration development has been observed. CONCLUSIONS: Combined SPE and GC-MS proved to be an efficient method to identify and quantify polar organic compounds in environmental samples. With respect to the concentrations measured in the present study, bisphenol A sometimes is the prominent compound in influent samples. Neither bisphenol A nor 4-tert-octylphenol or tech. 4-nonylphenol show seasonal variations. However, there was a significant general trend of decreasing concentrations of tech. 4-nonylphenol in influent and effluent samples from both WWTPs which probably reflects the implementing Directive 2003/53/EC. RECOMMENDATIONS AND PERSPECTIVES: Further research is needed to investigate whether the observed decrease of tech. 4-nonylphenol concentrations in German WWTPs since June 2004 will continue further on. The reason for the high effluent concentrations of bisphenol A in only a few samples has to be clarified in further research. The results from this study provide insight into the concentration development of the xenoestrogens bisphenol A, tech. 4-nonylphenol, and 4-tert-octylphenol in WWTPs in the time span between 2003 and 2005.