Sorption of polycyclic aromatic hydrocarbons (PAHs) to low and high density polyethylene (PE)

Background, aim, and scope

According to their high sorption capacity polyethylene (PE) passive samplers are often used for the analysis of polycyclic aromatic hydrocarbons (PAHs) in the aquatic environment. PE is also one of the primary synthetic polymers found in oceans, and sorption of PAHs to marine PE debris may determine PAH exposure and therefore hazards in marine ecosystems. Thus, an understanding of the sorption process is of great importance. In the present study, the sorption of several PAHs with different polarities to low density polyethylene (LDPE) and high density polyethylene (HDPE) was studied in order to improve our understanding of the influence of material properties on the Fickian diffusion of PAHs into PE.

Materials and methods

Batch sorption experiments were performed with aqueous solutions containing acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, and LPDE or HDPE pellets. Samples were shaken in the dark at 20?±?1°C for 16 time intervals within one week. Concentrations of PAHs were determined in the aqueous samples using solid-phase microextraction coupled with gas chromatography–mass spectrometry. The distribution coefficients (K _PE) between PE and water were estimated from different models reported in the literature. Kinetic sorption of the PAHs into the plastic pellets was described by a diffusion model based on Fick’s second law in spherical coordinates.

Results and discussion

A comparison between different models describing the equilibrium distribution of PAHs between PE and water revealed that the sorption equilibrium seemed to be driven by parameters other than, or in addition to, organic carbon. For both plastic types, diffusion coefficients decreased while the molecular weight of the PAHs increased which indicates a hindered diffusion through the matrix as a result of a larger molecule size. Higher diffusion coefficients were derived for LPDE than for HDPE indicating a greater sorption velocity for LPDE according to the lower polymer density.

Conclusions

Our results revealed that equilibrium time could be shortened during passive sampling as polymer membranes of lower density are used. In some areas, marine ecosystems may not be in equilibrium with respect to concentrations of organic contaminants and abundance of marine plastic debris. In such cases, different polymer densities should be taken into account in risk assessments.

     

Genotoxicity potential of a new natural formicide

Background, aim, and scope

Assessment of environmental impacts from pesticide utilization should include genotoxicity studies, where the possible effects of mutagenic/genotoxic substances on individuals are assessed. In this study, the genotoxicity profile of the new formicide Macex® was evaluated with two genotoxicity tests, namely, the micronucleus test with mouse bone marrow and Vicia faba, and a mutagenicity test using the Ames Salmonella assay.

Materials and methods

The bacterial reverse mutation test (Salmonella typhimurium strains TA97, TA98, TA100, TA102, and TA1535), the Vicia root tip and mouse micronucleus tests were conducted according to published protocols.

Results

In the range of the formicide Macex® concentrations tested from 0.06 to 1.0 g?L^?1 (or mgkg^?1 in the mouse test), no genotoxicity was observed in the prokaryotic or eukaryotic test organisms. However, at Macex® concentrations of 0.5 g?L^?1 and above a significant decrease in the mitotic index (P?≤?0.05) in the V. faba was observed. Micronucleus formation was likewise increased in the test organism at concentrations starting at 2.0 g?L^?1.

Conclusions

These data allow us to classify this natural formicide preparation as a product with no geno-environmental-impact when applied at recommended concentrations.

     

Heavy metal contamination in the vicinity of an industrial area near Bucharest

Background, aim, and scope

Heavy metals such as lead are well known to cause harmful health effects. Especially children are particularly susceptible to increased levels of lead in their blood. It is also a fact that lead concentration is increasing in the environment due to increased anthropogenic activity. The risk of heavy metal contamination is pronounced in the environment adjacent to large industrial complexes. In a combined case study, the environmental pollution by heavy metals was related to children’s health in the vicinity of an industrial area located 4 km south-east from Bucharest about 2 km east from the nearest town—Pantelimon. This site includes companies processing different, nonferrous solid wastes for recovery of heavy metals and producing different nonferrous alloys and lead batteries. In this paper, mainly the results of environmental sampling and analyses are summarized.

Materials and methods

Water, soil, and atmospheric deposition samples were collected from different locations within 3 km from the industrial area. For comparison, samples were also taken from Bucharest. Water samples were filtered (<0.45 μm), extracted by salpetric acid, and quantified by ICP-OES and ICP-MS. Soil samples were dried, sieved (<2 mm), extracted by aqua regia and analyzed by AAS. In order to quantify the atmospheric deposition, three kinds of permanently open collecting pots were used on nine different sites between August and November 2006.

Results

At most sampling locations, the heavy metal concentrations in soil decrease with increasing distance to the presumably major source of pollution. Highest heavy metal concentrations were found in 10–20 cm soil depths. There were also decreasing heavy metal concentrations for atmospheric deposition with increasing distance to the industrial site. In surface and groundwater samples, traces of zinc, copper and lead were detected.

Discussion

The heavy metal concentrations in soil were increased in the study area, mostly under legal action limits in low-concern areas (e.g., 1,000 mg Pb/kg dry soil), but often above action limits for high-concern areas (100 mg Pb/kg dry soil) such as populated areas. The soluble lead concentrations in water samples indicate a need for monitoring and assessing water quality in more detail. The results for atmospheric deposition showed increased dust precipitation and heavy metal loads in the study area compared to Bucharest. However, based on mass flow balance calculations, the actual atmospheric deposition of heavy metals must be much lower than it was in the past decades.

Conclusions

It was shown that highest lead values in water, soil and atmospheric deposition are rather to be found near the investigated industrial site than at the control sites in Bucharest. Our results correspond very well with results that show that children from Pantelimon have significantly increased lead concentrations in their blood compared to children in Bucharest. The increased lead contamination around the investigated industrial area is likely to have caused the increased exposure for children living in Pantelimon.

Recommendations and perspectives

In high-concern areas, such as found in populated areas, further measures have to be taken to avoid health risks for people living in these areas. The measures already taken to reduce emissions from the industrial site will help to avoid further increases in heavy metal concentrations. In areas with exceeded action limits, measures have to be taken as required by law. Detailed risk assessments could help to take necessary actions to protect public health in this area. The public should be informed about the potential hazards of eating plants grown in that area. Educational programs for schools, informing children about the contamination, should lead to a better understanding of environmental problems and a more sustainable behavior in the future.

     

Determination of polycyclic aromatic hydrocarbons in dry tea

Twenty-eight different tea samples sold in the United States were evaluated using high-performance liquid chromatography (HPLC) with fluorescence detection (FLD) for their contamination with polycyclic aromatic hydrocarbons (PAHs). Many PAHs exhibit carcinogenic, mutagenic, and teratogenic properties and have been related to several kinds of cancer in man and experimental animals. The presence of PAHs in environmental samples such as water, sediments, and particulate air has been extensively studied, but food samples have received little attention. Eighteen PAHs congeners were analyzed, with percentage recovery higher than 85%. Contamination expressed as the sum of the 18 analyzed PAHs was between 101 and 1337 μg/kg on dry mass and the average contents in all of the 28 examined samples was 300 μg/kg on dry mass. Seven of the congeners were found in all samples with wide ranges of concentrations as follows: fluorene (7–48 μg/kg), anthracene (1–31 μg/kg), pyrene (1–970 μg/kg), benzo(a)anthracene (1–18 μg/kg) chrysene (17–365 μg/kg), benzo(a)pyrene (1–29 μg/kg), and indeno(1,2,3-cd)pyrene (4–119 μg/kg). The two most toxic congeners benzo(a)pyrene and dibenzo(a,h)anthracene were found at high concentrations only in Earl Grey Twinnings, Earl Grey Harney& Sons Fine Teas, and Chai Ultra Spice Black Tea Twinnings. Six PAH congeners are considered as suspected carcinogens (U.S.EPA), formed the basis of the estimation of the toxic equivalent (TEQ), Chai Ultra-Spice Black Tea Twinnings had the highest TEQ (110.9) followed by two grey tea samples, Earl Grey Harney & Sons Fine Tea (57.7) and Earl Grey Twinnings (54.5). Decaffeinated grey teas had the lowest TEQs, decaffeinated Earl Grey Bigelow (9.4) and Green Tea Honey Lemon Decaffeinated Lipton (9.6).     

Evaluation of the giant reed (<Emphasis Type="Italic">Arundo donax</Emphasis>) in horizontal subsurface flow wetlands for the treatment of recirculating aquaculture system effluent

Introduction

Two emergent macrophytes, Arundo donax and Phragmites australis, were established in experimental subsurface flow, gravel-based constructed wetlands (CWs) receiving untreated recirculating aquaculture system wastewater.

Materials and methods

The hydraulic loading rate was 3.75 cm day^?1. Many of the monitored water quality parameters (biological oxygen demand [BOD], total suspended solids [TSS], total phosphorus [TP], total nitrogen [TN], total ammoniacal nitrogen [TAN], nitrate nitrogen [NO₃], and Escherichia coli) were removed efficiently by the CWs, to the extent that the CW effluent was suitable for use on human food crops grown for raw produce consumption under Victorian state regulations and also suitable for reuse within aquaculture systems.

Results and discussion

The BOD, TSS, TP, TN, TAN, and E. coli removal in the A. donax and P. australis beds was 94%, 67%, 96%, 97%, 99.6%, and effectively 100% and 95%, 87%, 95%, 98%, 99.7%, and effectively 100%, respectively, with no significant difference (p?>?0.007) in performance between the A. donax and P. australis CWs. In this study, as expected, the aboveground yield of A. donax top growth (stems + leaves) (15.0?±?3.4 kg wet weight) was considerably more than the P. australis beds (7.4?±?2.8 kg wet weight). The standing crop produced in this short (14-week) trial equates to an estimated 125 and 77 t ?ha^?1 year^?1 biomass (dry weight) for A. donax and P. australis, respectively (assuming that plant growth is similar across a 250-day (September–April) growing season and a single-cut, annual harvest).

Conclusion

The similarity of the performance of the A. donax- and P. australis-planted beds indicates that either may be used in horizontal subsurface flow wetlands treating aquaculture wastewater, although the planting of A. donax provides additional opportunities for secondary income streams through utilization of the energy-rich biomass produced.

     

Chemical,biological, and ecotoxicological assessment of pesticides and persistent organic pollutants in the Bahlui River,Romania

Background, aim, and scope

Current knowledge on environmental impacts of industrial activities in Romania, particularly persistent organic pollutants (POPs), indicates that environmental standards of the European Union are not systematically met. In our study area, additional sources of POPs are agriculture and domestic wastes. Very scarce information is available upon environmental contaminations and effects. In the present study, we investigated the chemical pollution and their eventual impact on the ecosystem by measuring POPs and by using biological indicators of pollution.

Materials and methods

The survey was carried out at six main sample sites along the Bahlui River. Sediments were chemically analysed for their content in polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs) and organochlorine pesticides (OCPs)—hexachlorocyclohexanes (HCHs) isomers and the dichlorodiphenyl trichloroethane (DDT) family. River water was biologically monitored at the level of phytoplankton and benthic invertebrates’ communities. Water samples from six locations have been analyzed for algal species composition and correspondence to various water quality indices. Biological samples have been taken from the same locations so as to calculate the macroinvertebrate indices. In the most polluted areas, as revealed by previous methods, toxicity was tested by exposing the green alga Pseudokirchneriella subcapitata and the cladoceran Daphnia magna to various dilutions of water sample.

Results

Important concentrations of POPs were identified only in sediments at river mouths (sites S5 and S6). Along the year, the sum of PCB concentrations ranged between 3 and 10 ng/g dw (S5), and between 4 and 26 ng/g dw (S6). Concentration of HCHs ranged between 0.4 and 3 ng/g dw (both S5 and S6) with a higher contribution of the gamma-HCH (30–70%), followed by beta-HCH (20–50%). The beta-HCH isomer was found at lower concentrations or even not detected in outer city sites. DDTs were found at higher concentrations than HCHs and ranged between 0.18 and 4 ng/g dw (S5) and between 0.56 and 18 ng/g dw (S6). The parent compound, p,p′-DDT, could be detected only in low concentrations (up to 5 ng/g dw) and contributed with less than 30% to the sum of DDTs in sediment. The principal contributors of the ΣDDTs in sediment were p,p′-DDE, and p,p′-DDD. The o,p′-DDD and DDT isomers were minor contributors to the sum of DDT. PAHs were found at higher concentrations than DDTs and ranged between 6 and 36 ng/g dw (S5) and between 36 and 155 ng/g dw (S6). Fluoranthene was predominant (up to 40%), followed by phenantrene (up to 30%), naphthalene (up to 35%), and benzo-(g,h,i)-perylene (up to 23%). The saprobity index and the diatom index increased from springs to river mouth, indicating a decrease in the water quality, but within the limits of moderate pollution. The saprobity index varied between 1.99 at spring to 2.70 at mouth. The diatom index varied from 3.48 to 3.14. The species’ richness in phytoplankton has a less clear pattern along the river, but in general, it appears to be negatively influenced by pollution. At the level of biological consumer species, the analyses of the macroinvertebrates confirm the situation and the tendency shown with algae. In addition, the Shannon–Wiener index, the Pielou evenness index, and the McNaughton dominance index indicate a peculiar pattern: invertebrate communities appear to a have a more stable structure along the river, with visible shifts at springs and at river mouths. Water toxicity testing indicates low toxicity of river waters around the city of Iasi, with two notable exceptions: the point pollution at the domestic wastewater treatment discharge and at the old open-air deposit of domestic solid wastes. Another important result was that tested algae appeared to be more readily affected than tested cladoceran: EC₅₀ (percent effluent) was 16 in algae and 28 in cladocerans. The slope of toxic effect was also much steeper in cladocerans (6) than in algae (1.8), which means that the toxic effect is more sudden on the tested invertebrates than on the tested algae.

Discussion

Pollutant concentrations reported herein are lower or similar than those reported for the sediments by earlier studies (RIZA 2000; Dragan et al., Int J Environ Anal Chem 86:833–842, 2006). Ratios of individual PAH compounds indicate important pyrolytic inputs and suggest that PAHs in the area are derived from the combustion of fossil fuels. Biologically, the waters appear to be beta-mesosaprobic towards alpha-mesosaprobic according to the saprobic index classification and undergo moderate pollution according to the diatom index classification. Water quality decreases from springs to river mouths. Algal species richness index has a less clear pattern along the river. Water toxicity is low, but certain sources of point pollution require increased attention.

Conclusions

The water quality is better than expected, probably because of the drop in pollution intensity following the collapse of local agricultural and industrial activity following the fall of communism in 1989. Nevertheless, further studies will be needed to confirm and refine our results. While this study draws no strident alarm, it appeals for high attention, particularly because the economic activity in the area is expected to increase.

Recommendations and perspectives

Future close monitoring will be necessary for insuring compliance with the Water Framework Directive, and for refining standards and understanding of the local situation, but with relevance for the wider international community. On the basis of the situation described in the present study, we recommend that future studies dedicate specific efforts to point pollution and effluent toxicity, particularly around the city of Iasi. For a better understanding of pollution and its effects, we recommend pursuing the type of multidisciplinary investigations proposed by the present study: chemical, ecotoxicological, and ecological. We also recommend that new methods should be developed and/or refined, like the empirical determination of partitioning coefficients in water and soils, process-based toxicity methods in ecotoxicological assessments, searching for interactions between pollution, producers, and consumers in aquatic ecosystems. We also recommend preference for cheaper survey methods, as these will be more applicable locally.

     

A preliminary investigation of the environmental impact of a thermal power plant in relation to PCB contamination

Purpose

The most significant application of polychlorinated biphenyls (PCBs) is in transformers and capacitors. Therefore, power plants are important suspected sources for entry of PCBs into the environment. In this context, the levels and distribution of PCBs in sediment, soil, ash, and sludge samples were investigated around Seyitömer thermal power plant, Kütahya, Turkey. Moreover, identity and contribution of PCB mixtures were predicted using the chemical mass balance (CMB) receptor model.

Methods

United States Environmental Protection Agency methods were applied during sample preparation, extraction (3540C), cleanup (3660B, 3665A, 3630C), and analysis (8082A).

Results

ΣPCB concentrations in the region ranged from not detected to 385 ng/g dry weight, with relatively higher contamination in sediments in comparison to soil, sludge, and ash samples collected from around the power plant. Congener profiles of the sediment and soil samples show penta-, hexa-, and hepta-chlorobiphenyls as the major homolog groups. The results from the CMB model indicate that PCB contamination is largely due to Clophen A60/A40 and Aroclor 1254/1254(late)/1260 release into the sediment and sludge samples around the thermal power plant.

Conclusions

Since there are no other sources of PCBs in the region and the identity of PCB sources estimated by the CMB model mirrors PCB mixtures contained in transformers formerly used in the plant, the environmental contamination observed especially in sediments is attributed to the power plant. Release of PCBs over time, as indicated by the significant concentrations observed even in surface samples, emphasizes the importance of the need for better environmental management.

     

Environmental exposure of pharmaceuticals and musk fragrances in the Somes River before and after upgrading the municipal wastewater treatment plant Cluj-Napoca,Romania

Background, aim, and scope

Pharmaceutically active substances are a class of emerging contaminants, which has led to increasing concern about potential environmental risks. After excretion, substantial amounts of unchanged pharmaceuticals and their metabolites are discharged into domestic wastewaters. The absence of data on the environmental exposure in Eastern Europe is significant, since use patterns and volumes differ from country to country. In Romania, the majority of wastewater, from highly populated cities and industrial complex zones, is still discharged into surface waters without proper treatment or after inefficient treatment. In respect to this, it is important to determine the environmental occurrence and behavior of pharmaceuticals and personal care products (PPCPs) in wastewaters and surface waters. The objective of the present study was to investigate the occurrence of selected PPCPs during the transport in the Somes River by mass flow analysis before and after upgrading a municipal wastewater treatment plant (WWTP) in Cluj-Napoca, which serves 350,000 inhabitants and is the largest plant discharging into the Somes River. The concentrations of PPCPs at Cluj-Napoca can be correlated with the high population and a high number of hospitals located in the catchment area leading to higher mass flows. The results of this study are expected to provide information, with respect to the Romanian conditions, for environmental scientists, WWTP operators, and legal authorities. The data should support the improvement of existing WWTPs and implementation of new ones where necessary and, therefore, minimize the input of contaminants into ambient waters.

Materials and methods

The PPCPs were selected on the basis of consumption at the regional scale, reported aquatic toxicity, and the suitability of the gas chromatography/mass spectrometry (GC/MS) method for the determination of the compounds at trace levels. The studied PPCPs, caffeine (stimulant), carbamazepine (antiepileptic), pentoxifylline (anticoagulant), cyclophosphamide (cytostatic), ibuprofen (analgesic), and galaxolide (musk fragrance), were determined in samples of the Somes River. The analytes were enriched by solid-phase extraction and subsequently determined by GC/MS. Caffeine, pentoxifylline, and galaxolide were determined underivatized, whereas the acidic pharmaceuticals carbamazepine, cyclophosphamide, and ibuprofen were determined after derivatization with N-methyl-N-(trimethylsilyl)-trifluoroacetamide.

Results and discussion

The concentrations in the Somes River varied from below 10 ng/L up to 10 μg/L. A substantial decrease of the exposure in the Somes River could be observed due to the upgrade of the municipal WWTP in Cluj-Napoca. The loads in the river stretch between Cluj-Napoca and Dej (Somes Mic) varied strongly: caffeine (400–2,000 g/day), carbamazepine (78–213 g/day), galaxolide (140–684 g/day), ibuprofen (84–108 g/day). After the upgrade of the WWTP Cluj-Napoca, the concentrations in the Somes of caffeine, pentoxifylline, cyclophosphamide, galaxolide, and tonalide were significantly reduced (over 75%). One might be cautious comparing both studies because the relative efficiency of the WWTP’s removal of PPCP was not evaluated. However, the significantly lower concentrations of most compounds after the upgrade of the WWTP Cluj-Napoca allow one to infer that the technical measures at the source substantially reduced inputs of contaminants to the receiving river. Dej loads of the poorly biodegradable substance carbamazepine increased by a factor of 2–3 as a result of wastewater discharges into the river. The disproportionate increase in caffeine loads by a factor of 4 below Cluj-Napoca indicates inputs of untreated wastewater from the Somes Mare due to the discharge of untreated wastewater derived from Bistrita, Nasaud, and Beclean (115,000 inhabitants).

Conclusions

The relative contribution of treated and untreated wastewater in surface water might be assessed by measuring chemical markers. Recalcitrant pharmaceuticals like carbamazepine are suitable as chemical markers for estimating the relative contribution of wastewater in surface water. The easily degradable caffeine might be a good indicator for raw sewage and hardly treated wastewaters.

Recommendations and perspectives

Municipal WWTPs have the potential of a significant contribution in reducing the load of contaminants to ambient waters. The efficiency of the wastewater treatment in Cluj-Napoca improved considerably after the upgrade of the WWTP. Therefore, it is crucial that several WWTPs must be implemented or improved in the Somes Valley Watershed in order to reduce the discharge of contaminants in the Somes River from these point sources.

     

Response to W. Kramer: The human sex odds at birth after the atmospheric atomic bomb tests,after Chernobyl,and in the vicinity of nuclear facilities: comment (doi:10.1007/s11356-011-0644-8)

Introduction

This paper is in response to criticism of our article “The human sex odds at birth after the atmospheric atomic bomb tests, after Chernobyl, and in the vicinity of nuclear facilities” published in Environ Sci Pollut Res 18(5):697–707, 2011.

Methods

Our findings and methods concerning the disturbed human sex odds at birth have been criticized in this journal for being artifacts of data mining, that the concept of statistical significance was misunderstood, and that confounding factors have not been accounted for. Here, we show that this criticism has no basis. We applied well-established statistical methods to large official data sets, and confounding is less important at the level of secular sex odds trends in aggregated annual figures from countries or continents.

Results and conclusions

Moreover, our results are strengthened by recent findings concerning increased infant death sex odds in Germany and increased Down syndrome prevalence at birth across Europe after Chernobyl. Prompted by our studies, an official investigation in Lower Saxony, Germany, by the “Niedersächsisches Landesgesundheitsamt (NLGA)” confirmed our observation of severely escalated sex odds within 40 km distance from the nuclear storage site in Gorleben, Germany.

     

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.

     

Heavy metals in untreated/treated urban effluent and sludge from a biological wastewater treatment plant

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.

     

Occurrence and temporal variations of TMDD in the river Rhine,Germany

Background, aim, and scope

The chemical substance 2,4,7,9-tetramethyl-5-decyne-4,7-diol (TMDD) is a non-ionic surfactant used as an industrial defoaming agent and in various other applications. Its commercial name is Surynol 104® and the related ethoxylates are also available as Surfynol® 420, 440, 465 and 485 which are characterized by different grades of ethoxylation of TMDD at both hydroxyl functional groups. TMDD and its ethoxylates offer several advantages in waterborne industrial applications in coatings, inks, adhesives as well as in paper industries. TMDD and its ethoxylates can be expected to reach the aquatic environment due its widespread use and its physico-chemical properties. TMDD has previously been detected in several rivers of Germany with concentrations up to 2.5?µg/L. In the United States, TMDD was also detected in drinking water. However, detailed studies about its presence and distribution in the aquatic environment have not been carried out so far. The aim of the present study was the analysis of the spatial and temporal concentration variations of TMDD in the river Rhine at the Rheingütestation Worms (443.3 km). Moreover, the transported load in the Rhine was investigated during two entire days and 7 weeks between November 2007 and January 2008.

Materials and methods

The sampling was carried out at three different sampling points across the river. Sampling point MWL1 is located in the left part of the river, MWL2 in the middle part, and MWL4 in the right part. One more sampling site (MWL3) was run by the monitoring station until the end of 2006, but was put out of service due to financial constrains. The water at the left side of the river Rhine (MWL1) is influenced by sewage from a big chemical plant in Ludwigshafen and by the sewage water from this city. The water at the right side of the river Rhine (MWL4) is largely composed of the water inflow from river Neckar, discharging into Rhine 14.9 km upstream from the sampling point and of communal and industrial wastewater from the city Mannheim. The water from the middle of the river (MWL2) is largely composed of water from the upper Rhine. Water samples were collected in 1-L bottles by an automatic sampler. The water samples were concentrated by use of solid-phase extraction (SPE) using Bond Elut PPL cartridges and quantified by use of gas chromatography-mass spectrometry (GC-MS). The quantification was carried out with the internal standard method. Based on these results, concentration variations were determined for the day profiles and week profiles. The total number of analyzed samples was 219.

Results

The results of this study provide information on the temporal concentration variability of TMDD in river Rhine in a cross section at one particular sampling point (443.3 km). TMDD was detected in all analyzed water samples at high concentrations. The mean concentrations during the 2 days were 314 ng/L in MWL1, 246 ng/L in MWL2, and 286 ng/L in MWL4. The variation of concentrations was low in the day profiles. In the week profiles, a trend of increasing TMDD concentrations was detected particularly in January 2008, when TMDD concentrations reached values up to 1,330 ng/L in MWL1. The mean TMDD concentrations during the week profiles were 540 ng/L in MWL1, 484 ng/L in MWL2, and 576 ng/L in MWL4. The loads of TMDD were also determined and revealed to be comparable in all three sections of the river. The chemical plant located at the left side of the Rhine is not contributing additional TMDD to the river. The load of TMDD has been determined to be 62.8 kg/d on average during the entire period. By extrapolation of data obtained from seven week profiles the annual load was calculated to 23 t/a.

Discussion

The permanent high TMDD concentrations during the investigation period indicate an almost constant discharge of TMDD into the river. This observation argues for effluents of municipal wastewater treatment plants as the most likely source of TMDD in the river. Another possible source might be the degradation of ethoxylates of TMDD (Surfynol® series 400), in the WWTPs under formation of TMDD followed by discharge into the river. TMDD has to be considered as a high-production-volume (HPV) chemical based on the high concentrations found in this study. In the United States, TMDD is already in the list of HPV chemicals from the Environmental Protection Agency (EPA). However, the amount of TMDD production in Europe is unknown so far and also the biodegradation rates of TMDD in WWTPs have not been investigated.

Conclusions

TMDD was found in high concentrations during the entire sampling period in the Rhine river at the three sampling points. During the sampling period, TMDD concentrations remained constant in each part of the river. These results show that TMDD is uniformly distributed in the water collected at three sampling points located across the river. ‘Waves’ of exceptionally high concentrations of TMDD could not be detected during the sampling period. These results indicate that the effluents of WWTPs have to be considered as the most important sources of TMDD in river Rhine.

Recommendations and perspectives

Based also on the occurrence of TMDD in different surface waters of Germany with concentrations up to 2,500 ng/L and its presence in drinking water in the USA, more detailed investigations regarding its sources and distribution in the aquatic environment are required. Moreover, the knowledge with respect to its ecotoxicity and its biodegradation pathway is scarce and has to be gained in more detail. Further research is necessary to investigate the rate of elimination of TMDD in municipal and industrial wastewater treatment plants in order to clarify the degradation rate of TMDD and to determine to which extent effluents of WWTPs contribute to the input of TMDD into surface waters. Supplementary studies are needed to clarify whether the ethoxylates of TMDD (known as Surfynol 400® series) are hydrolyzed in the aquatic environment resulting in formation of TMDD similar to the well known cleavage of nonylphenol ethoxylates into nonylphenols. The stability of TMDD under anaerobic conditions in groundwater is also unknown and should be studied.

     

A source classification framework supporting pollutant source mapping,pollutant release prediction,transport and load forecasting,and source control planning for urban environments

Purpose

Implementation of current European environmental legislation such as the Water Framework Directive requires access to comprehensive, well-structured pollutant source and release inventories. The aim of this work was to develop a Source Classification Framework (SCF) ideally suited for this purpose.

Methods

Existing source classification systems were examined by a multidisciplinary research team, and an optimised SCF was developed. The performance and usability of the SCF were tested using a selection of 25 chemicals listed as priority pollutants in Europe.

Results

The SCF is structured in the form of a relational database and incorporates both qualitative and quantitative source classification and release data. The system supports a wide range of pollution monitoring and management applications. The SCF functioned well in the performance test, which also revealed important gaps in priority pollutant release data.

Conclusions

The SCF provides a well-structured approach for European pollutant source and release classification and management. With further optimisation and demonstration testing, the SCF has the potential to be fully implemented throughout Europe.

     

Historical record of mercury contamination in sediments from the Babeni Reservoir in the Olt River,Romania

Background, aim and scope

Mercury (Hg) is a ubiquitous and hazardous contaminant in the aquatic environment showing a strong biomagnification effect along the food chain. The most common transfer path of Hg to humans is contaminated fish consumption. In severely exposed humans, Hg poisoning may lead to damage in the central nervous system. Thus, it is important to examine current and past contamination levels of Hg in aquatic milieu. The Olt River is the largest Romanian tributary of the Danube River. The use of Hg as an electrode in a chlor-alkali plant contributed to the contamination of the aquatic environment in the Rm Valcea region. The purpose of this study was to compare the current state of Hg contamination with the past contamination using a historical record obtained from a dated sediment core from one of the Olt River reservoirs (Babeni) located downstream from the chlor-alkali plant. To our knowledge, no published data on Hg contamination in this region are available. The Babeni Reservoir was selected for this study because it is situated downstream from the chlor-alkali plant, whilst the other reservoirs only retain the pollutants coming from the upstream part of the watershed. Preliminary analyses (unpublished) showed high Hg concentrations in the surface sediment of the Babeni Reservoir. One core was taken in the upstream Valcea Reservoir to provide a local background level of Hg concentrations in sediments.

Results and discussion

Sediment texture was uniform in the cores from both reservoirs. Laminated sediment structure, without any obvious discontinuities, was observed. Hg concentrations in the sediment core from the Valcea Reservoir were low and constant (0.01–0.08 mg/kg). In Babeni Reservoir sediments, Hg concentrations were very high in the deeper core section (up to 45 mg/kg in the longest core) and decreased to lower concentrations toward the top of the cores (1.3–2.4 mg/kg). This decrease probably reflects technological progress in control of emissions from the Hg-cell-based chlor-alkali industry. Two strong peaks could be distinguished in older sediments. The mean rate of sedimentation (5.9 cm/year) was calculated from the depth of the ¹³⁷Cs Chernobyl peak. This was in good agreement with the sedimentation rate estimated at this site from a bathymetric study. Assuming a constant sedimentation rate, the two Hg peaks would reflect two contamination events in 1987 and 1991, respectively. However, it is also possible that the two peaks belong to the same contamination event in 1987 but were separated by a sediment layer richer in sand and silt. This layer had a low Hg concentration, which can be interpreted as a mass deposition event related to a major flood bringing Hg-free sediments.

Conclusions

Whilst the chlor-alkali plant partly switched to a cleaner technology in 1999, no obvious decrease of Hg concentrations was observed in recent decade. Results from the sediment core reflected the historical trend of Hg release from the chlor-alkali plant, revealed important contamination episodes and confirmed a legacy of contamination of Hg in recent sediments even if the concentrations of Hg decreased toward the surface due to a more efficient emission control.

Recommendations and perspectives

Although the Hg concentrations in Babeni Reservoir sediments were extremely high in the late eighties and they remain one order of magnitude higher in the surface sediments than in sediments from the upstream reservoir, little is known about the transfer of Hg to the biota and human population. Our initial measurements indicate the presence of monomethyl-Hg (MMHg) in pore water, but further studies are necessary to evaluate fluxes of MMHg at the sediment–water interface. Samples of fish and hair from various groups of the local population were recently collected to evaluate the potential hazard of Hg contamination to human health in the Rm Valcea region.

     

Importance of exposure route for behavioural responses in <Emphasis Type="Italic">Lumbriculus variegatus</Emphasis> Müller (Oligochaeta: Lumbriculida) in short-term exposures to Pb

Goal, Scope and Background

Lumbriculus variegatus Müller (Oligochaeta), a common freshwater sediment-dweller, has frequently been used in toxicokinetic studies, although has been less used in ecotoxicity tests.

Methods

For the first time the Multispecies Freshwater Biomonitor® (MFB) was applied in a short-term whole-sediment toxicity test. The MFB automatically and quantitatively recorded the spontaneous locomotory behaviour of Lumbriculus variegatus in exposures with two compartments, water and sediment. The study questioned, whether the animals altered their locomotion depending on the compartment which was spiked with lead (Pb).

Results and Discussion

As in the exposures to Pb-contaminated water/clean sediment, the animals exposed to Pb-contaminated sediment/clean water showed higher activities in intermediate Pb-concentrations. This indicates, that spontaneous locomotory activity is affected by Pb-concentrations at sublethal levels regardless of whether the Pb-concentration is found in the water or in the sediment, because these animals use both environmental compartments simultaneously. However, within the same Pb-levels, the animals showed higher locomotory activity in contaminated water compared with contaminated sediment. This indicates a possible tendency to withdraw from (‘avoidance’) contaminated water into the clean sediment compartment, whereas there was no withdrawal from contaminated sediment into clean water. The latter might be explained by the fact that withdrawal from sediment to water might increase the risk of predation and drift in nature, whereas retracting to sediment might provide shelter.

Conclusions

The study showed that spontaneous locomotory responses of L. variegatus to Pb depend on whether the water or sediment is contaminated. The study also concluded that the Multispecies Freshwater Biomonitor® can be applied effectively in sediment toxicity testing.

Recommendations and Perspectives

More emphasis should be given to the interactions of water/sediment in sediment ecotoxicity tests to better simulate field conditions and increase ecological realism in risk assessment, especially as quantitative recording methods exisit.

     

The Rhine red,the fish dead—the 1986 Schweizerhalle disaster,a retrospect and long-term impact assessment

Background

The November 1, 1986 fire at a Sandoz Ltd. storehouse at Schweizerhalle, an industrial area near Basel, Switzerland, resulted in chemical contamination of the environment. The storehouse, which was completely destroyed by the fire, contained pesticides, solvents, dyes, and various raw and intermediate materials. The majority of the approximately 1,250 t of stored chemicals was destroyed in the fire, but large quantities were introduced into the atmosphere, into the Rhine River through runoff of the fire-fighting water, and into the soil and groundwater at the site. The chemicals discharged into the Rhine caused massive kills of benthic organisms and fish, particularly eels and salmonids. Public and private reaction to the fire and subsequent chemical spill was very strong. This happened only a few months after the Chernobyl accident and it destroyed the myth of immunity of Switzerland regarding such catastrophes.

Aim

This article reviews the damaging events of November 1986 and aims at striking stock two decades later.

Results and discussion

In the aftermath of this once-per-century accident, the aim was to obtain new knowledge for the environmental sciences and to achieve progress for water pollution control issues. The following themes are discussed: mitigation measures by the chemical industry and by the governmental authorities, activities of environmental protection organizations, chemical and biological monitoring, alert organization, ecological damages, ecotoxicological effects assessment, recovery and alteration of the river biology, return of the salmon, drinking water supplies, research programs, education of environmental scientists, and visions for the future.

Conclusions

The catastrophic pollution of the Rhine in November 1986 has triggered by the openly visible damages of the river biology that subsequently significant progress was made towards the prevention of such environmental catastrophes. The crucial risk reduction measures in the chemical industry, legal regulations and controls as well as chemical and biological monitoring of the river water quality were substantially improved. Politics and chemical industry have learned their lectures and have proceeded accordingly.

Recommendations

A drastic acute contamination, as it has happened at Schweizerhalle in 1986, is clearly recognizable by the toxic effects, which it is causing. This can lead to mitigation activities, which are positive considering a long-term perspective. However, the less obvious effects of chronic water pollution should receive more attention as well as the ongoing alteration of the biocenosis. A high water quality must be demanded for using water from the Rhine to produce drinking water. In that context, micropollutants should also be considered and particular attention should be payed to emerging contaminants.

Perspectives

The big chemical storehouse fire of 1986 had an important impact on the transboundary cooperation and has improved the willingness for international cooperation. The fire catastrophe of Schweizerhalle has triggered many activities in particular for the Rhine River. Overall, the effects are positive on the basis of a long-term perspective. The applied whole basin approach gives an example on a global basis for other river systems, which still are more heavily polluted.

     

Genotoxicity of oxy-PAHs to Japanese medaka (Oryzias latipes) embryos assessed using the comet assay

Polycyclic aromatic hydrocarbons (PAHs) have long been recognized as important environmental toxicants. Despite a plethora of information on the fate and effects of parent PAHs, relatively little is known about the environmental fate and toxicity of ketone- and quinone-substituted PAH oxidation products (termed oxy-PAHs), particularly in the aquatic environment. This study begins to fill that gap using embryos of the Japanese medaka (Oryzias latipes) as a model species. The genotoxic potential of two environmentally relevant oxy-PAHs, acenaphthenequinone and 7,12-benz[a]anthracenquinone, was assessed using the comet assay. We found that both oxy-PAHs could cause significant increases in DNA damage after only 48 h of exposure at the lowest concentrations tested (5 μg/L). Comparisons of the genotoxic potential between these oxy-PAHs and their corresponding parent PAHs (acenaphthene and benz[a]anthracene) and a well-known mutagenic PAH, benzo[a]pyrene, indicated similar potencies among all five of these compounds, particularly after longer (7 day) exposures. This study demonstrates the mutagenic potential of oxy-PAHs to an in vivo fish embryo model and points out the need for further study of their environmental occurrence and biologic effects.     

Ecotoxicological research and related legislation in Serbia

Introduction

Several hot spots of severe freshwater pollution and sediment contamination (mostly heavy metals, polyaromatic hydrocarbons and polychlorinated biphenyls) have been identified in Serbia as the consequence of outdated environmental legislation, negligible amounts of properly treated waste waters and accidental spills.

Discussion

Since ecotoxicological methods have never been incorporated into risk assessment procedures, mandatory effluent discharge or ambient water monitoring programmes, ecotoxicological research, based on bioaccumulation studies, conventional ecotoxicological tests and, recently, biomarkers of exposure and effect have been restricted to independent small- to medium-scale studies, conducted, basically, to confirm, underline or oppose the results of chemical-based monitoring and to lament on inadequate environmental regulations/policy and management practice. Although hot and unresolved ecotoxicological problems still remain beyond the reach of ecotoxicological research currently conducted in Serbia, or are tackled only sporadically, it is to be expected that on-going research and institutional capacity building should, hopefully, increase the competence and competitiveness of scientific community and speed up the process of harmonisation of national environmental legislation and policy with European Union.

     

Metals in biomass

Background, Aim and Scope

Metal ions generally share the ability/tendency of interacting with biological material by forming complexes, except possibly for the heavy alkali metals K, Rb and Cs. This is unrelated to the metals being either essential for sustaining life and its reproduction, apparently insignificant for biology, although perhaps undergoing bioconcentration or even being outright toxic, even at low admission levels. Yet, those different kinds of metal-biomass interactions should in some way depend on properties describing coordination chemistries of these very metals. Nevertheless, both ubiquitously essential metals and others sometimes used in biology should share these properties in numeric terms, since it can be anticipated that they will be distinguished from nonessential and/or toxic ones. These features noted above include bioconcentration, the involvement of metal ions such as Zn, Mg, Cu, Fe, etc. in biocatalysis as crucial components of metalloenzymes and the introduction of a certain set of essential metals common to (almost) all living beings (K, Mg, Mo, Mn, Fe, Cu and Zn), which occurred probably very early in biological evolution by ‘natural selection of the chemical elements’ (more exactly speaking, of the metallomes).

Materials and Methods

The approach is semiempirical and consists of three consecutive steps: 1) derivation of a regression equation which links complex stability data of different complexes containing the same metal ion to electrochemical data pertinent to the (replaced) ligands, thus describing properties of metal ions in complexes, 2) a graphical representation of the properties-two typical numbers c and x for each metal ion-in some map across the c/x-space, which additionally contains information about biological functions of these metal ions, i.e. whether they are essential in general (e.g. Mg, Mn, Zn) or, for a few organisms of various kinds (e.g. Cd, V), not essential (e.g. rare earth element ions) or even generally highly toxic (Hg, U). It is hypothesized that, if coordination properties of metals control their biological ‘feasibility’ in some way, this should show up in the mappings (one each for mono and bidentate-bonding ligands). 3) eventually, the regression equation produced in step 1) is inverted to calculate complex stabilities pertinent to biological systems: 3a) complex stabilities are mapped for ligands delivered to soil (-water) by green plants (e.g. citrate, malate) and fungi and, compared to their unlike selectivities and demands of metal use (photosynthesis taking place or not), 3b) the evolution of the metallome during late chemical evolution is reconstructed.

Results

These maps show some ‘window of essentiality’, a small, contrived range/area of c and x parameters in which essential metal ions gather almost exclusively. c and x thus control the possibility of a metal ion becoming essential by their influencing details of metal-substrate or (in cases of catalytic activities) metal-product interactions. Exceptions are not known to be involved in biocatalysis anyhow.

Discussion

Effects of ligands secreted, e.g. from tree roots or agaric mycelia to the soil on the respective modes (selectivities) of metal bioconcentration can be calculated by the equation giving complex stability constants, with obvious ramifications for a thorough, systematic interpretation of biomonitoring data. Eventually, alterations of C, N and P-compounds during chemical evolution are investigated — which converted CH₄ or CO₂, N₂ and other non-ligands to amino acids, etc., for example, with the latter behaving as efficient chelating ligands: Did they cause metal ions to accumulate in what was going to become biological matter and was there a selectivity, a positive bias in favour of nowessential metals (see above) in this process? Though there was no complete selectivity of this kind, neither a RNA world in which early ribozymes effected most of biocatalysis, nor a paleoatmosphere containing substantial amounts of CO could have paved the way to the present biochemistry and metallomes.

Conclusions

This way of reasoning provides a causal account for abundance distributions described earlier in the Biological System of Elements (BSE; Markert 1994, Fränzle &; Markert 2000, 2002). There is a pronounced change from chemical evolution, where but few transformations depended on metal ion catalysis to biology.

Recommendations and Perspectives

The application of this numerical approach can be used for modified, weighted evaluation of biomonitoring analytical data, likewise for the prediction of bioconcentration hazards due to a manifold of metal ions, including organometallic ones. This is relevant in ecotoxicology and biomonitoring. In combining apoproteins or peptides synthesized from scratch for purposes of catalysing certain transformations, the map and numerical approaches might prove useful for the selection of central ions which are even more efficient than the ‘natural’ ones, like for Co²⁺ in many Zn enzymes.

     

Antioxidative responses of duckweed (<Emphasis Type="Italic">Lemna minor</Emphasis> L.) to short-term copper exposure

Goal, Scope and Background

Elevated concentrations of copper in the environment result in accumulation of the metal in plants and cause an increase in reactive oxidative species (ROS). The first response to elevated amounts of ROS is increased levels of enzymatic and non-enzymatic antioxidants that reduce oxidative stress. The aim of our study was to evaluate the early stages of antioxidative responses to the low copper concentrations usually present in moderately polluted environments. In addition, some other parameters were examined to evaluate the effect of copper on plants.

Methods

Duckweed (Lemna minor L.) was exposed to different concentrations of copper sulphate for up to 24 hours. Glutathione concentration and enzymatic activities of catalase, guaiacol peroxidase and glutathione reductase were measured spectrophotometrically. Additionally, delayed and prompt chlorophyll fluorescence was measured by luminometry and fluorometry, respectively. The accumulation of copper in plants exposed for 24 hours to various concentrations of copper sulphate was measured by flame atomic absorption spectrophotometry.

Results

The treatment of plants with copper sulphate resulted in an immediate decrease of the glutathione pool, which was replenished after 24 hours at CuSO₄ concentrations lower than 2 μM. Higher CuSO₄ concentrations caused a decrease of reduced glutathione. The responses of the antioxidant enzymes glutathione reductase, guaiacol peroxidase and catalase to CuSO₄ differed during the first six hours of exposure, but their enzyme activities all increased after 24 hours of exposure. All these enzymes displayed biphasic activity curves with maximum values between 0.5 μM and 1 μM CuSO₄. The response of guaiacol peroxidase was the most pronounced and statistically significantly specific and that of catalase the least. Delayed chlorophyll fluorescence decreased after exposure to 1 μM CuSO₄, but no significant effect on maximum quantum yield of photosystem II (Fv/Fm) was observed. L. minor accumulated relatively high concentrations of copper. The accumulation rate was higher at lower concentrations of copper in the test medium (up to 2 μM CuSO₄) than at concentrations above 2 μM CuSO₄.

Discussion

One of the most pronounced antioxidative responses to copper exposure was modified levels of oxidized and reduced forms of glutathione. The decrease of the glutathione pool is most probably coupled with induced production of phytochelatins. Antioxidative enzymes showed the biphasic enzyme activity characteristic of stress response. Guaiacol peroxidase exhibited the greatest significant increase of activity, even at higher CuSO₄ concentrations at which the activity of catalase and glutathione reductase dropped. The intensity of delayed chlorophyll fluorescence decreased, indicating reduced photosynthesis of plants under stress. All the measured parameters showed that plants respond to even low copper concentrations very soon after exposure. The accumulation rate of copper in duckweed tissues indicates that L. minor is an accumulator species.

Conclusions

The synchronized and prompt inducibility of antioxidants indicates their involvement in a general plant defence strategy for coping with metal-induced oxidative stress. Glutathione concentration and guaiacol peroxidase activity were found to be the most sensitive of the early indicators of exposure to copper concentrations present in polluted water bodies.

Recommendation and Perspectives

The experimental design of the present study allowed us to compare the sensitivity of various methods and parameters for detecting plant responses to heavy metal-induced oxidative stress. The level of glutathione and the enzyme activities of guaiacol peroxidase and glutathione reductase could be used as a rapidly determined early warning system in toxicity studies.