Biomedical subjects

Rolf Altenburger

Publications and source records attributed to Rolf Altenburger.

18 recordsLinked to original sources

PubMed2006

Development and validation of a new fluorescence-based bioassay for aquatic macrophyte species.

Bioassays with unicellular algae are frequently used as ecotoxicological test systems to evaluate the toxicity of contaminated environmental samples or chemicals. In contrast, aquatic macrophyte test systems are still rarely used as they are laborious to handle because species exhibit distinct ecological requirements. The aim of this study was to establish a fast and reproducible measuring system for aquatic macrophyte species to overcome those limitations for use. Thus, a newly developed pulse-amplitude modulated chlorophyll fluorometer (Imaging-PAM) was applied as an effect detection in short-term bioassays with aquatic macrophyte species. This multiwell-plate-based measuring device enables the incubation and measurement of up to 24 samples in parallel. The Imaging-PAM was used (i) to establish and validate the sensitivity of the test systems to three Photosystem II (PSII) inhibitors (atrazine, prometryn, isoproturon), (ii) to compare the test systems with established biotests for macrophytes and (iii) to define necessary time scales in aquatic macrophyte testing. The results showed that fluorescence-based measurements with the Imaging-PAM allow rapid and parallel analysis of large amounts of aquatic macrophyte samples and of toxicants effects of the PSII inhibitors tested on aquatic macrophytes. Measurements revealed a good correlation between obtained median effective concentrations (EC50s) for the new and the established biotest systems. Hence, the Imaging-PAM measuring device is a promising tool to allow fast chemical effect screening for high amounts of samples with little time and material and thus offers scope for high-throughput biotesting using aquatic macrophyte species.

Atrazine↗

PubMed2006

Phytotoxicity assessment of diclofenac and its phototransformation products.

The occurrence of pharmaceuticals in the environment is an emerging issue. Several studies observed that the non-steroidal anti-inflammatory drug diclofenac is ubiquitously present in most of the surveyed surface waters, worldwide. Phototransformation of diclofenac was reported from laboratory assays as well as in natural water systems, raising the question of possible adverse effects of the phototransformation products of diclofenac to aquatic organisms. In this study the phytotoxicity of diclofenac exposed to natural sunlight was evaluated using synchronized cultures of the unicellular chlorophyte Scenedesmus vacuolatus. Diclofenac dissolved in ultra-pure water at 50 mg L(-1) was exposed to natural midsummer sunlight for a maximum of 145 h. Twice a day subsamples were taken for chromatography and parallel phytotoxicity assessment. Inhibition of algal reproduction of the initial diclofenac solution was in the mg L(-1) range indicating no specific toxicity of diclofenac towards S. vacuolatus. Fast degradation of diclofenac was observed with half lives between 3.3 and 6.4 h during the first and the third day of exposure, respectively. Phytotoxicity increased after 3.5 h of exposure of diclofenac to sunlight and showed a maximum of sixfold enhanced toxicity after 53 h of exposure to sunlight. Several phototransformation products were found during the experiment. The time courses of the relative concentration of three transformation products significantly correlated with enhanced phytotoxicity during the experiment. This indicates a high toxicity potential of phototransformation products of diclofenac at concentration levels that may come close to environmental concentrations of residual diclofenac after degradation. We conclude that toxicity assessment of phototransformation products should be included in the risk assessment of pharmaceuticals in the environment.

Chromatography, High Pressure Liquid↗

PubMed2006

On the mode of action of N-phenyl-2-naphthylamine in plants.

N-phenyl-2-naphthylamine, a sediment contaminant previously identified as a major toxicant of site-specific importance was investigated for its mode of toxic action. From short-term bioassays with daphnids, fish eggs, bacteria, and algae it appears that this compound has specific phytotoxic properties at concentrations below 100 microg/L, which cannot be explained assuming an unspecific narcosis type of action in plants. Also, hydroxy-, nitro-, and methylderivatives show clear excess toxicity as compared to baseline toxic effects. Of several plant-specific growth and development processes investigated, only photosynthesis could be demonstrated to be affected at short exposure times and low concentrations. Disturbance of primary photosynthetic reactions such as oxygen evolution and fluorescence quenching, however, becomes only apparent after 2-3 h of exposure, which is in sharp contrast to known specific inhibitors targeting processes such as electron transport or ATP production. This, and concentration-time-effect modeling lead to the suggestion that N-phenyl-2-naphthylamine acts intracellular as a reactive compound in cell membranes producing irreversible, and thus cumulative, damage over time in algae. The effects may become first apparent in membrane-rich compartments such as the algal chloroplast.

2-Naphthylamine↗

PubMed2005

Modeling photoinduced algal toxicity of polycyclic aromatic hydrocarbons.

The influence of light conditions on the toxicity of polycyclic aromatic hydrocarbons (PAHs) to different organisms has long been recognized. The aim of this study was to investigate whether previously proposed models can be used to qualitatively and quantitatively predict photoinduced toxicity to the green algae Scenedesmus vacuolatus. For this purpose 14 different PAH compounds were tested under three different light conditions for their effects on the algae reproduction. Illumination conditions comprised standard algae growth light, simulated sunlight aiming to mimic environmental light conditions, and UV-filtered light in order to minimize light influence on PAH toxicity. Models proposed for the prediction of photoinduced toxicity were modified in order to account for different exposure conditions and toxic endpoints used in the bioassay. The results of this study show that the gap between the highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) proposed as an indication for photoinduced toxicity to Daphnia magna can be used as a qualitative indication of a potential photoinduced toxicity to the green algae Scenedesmus vacuolatus. The impact of light conditions on PAH toxicity can be quantified by a linear model which allows the estimation of the ED50 of each compound from the amount of absorbed photons and an empirically determined relative phototoxic efficacy value of the compound.

Chlorophyta↗

PubMed2005

Chemical and ecotoxicological assessment of polycyclic aromatic hydrocarbon--contaminated sediments of the Niger Delta, Southern Nigeria.

The extent of environmental contamination and sources of polycyclic aromatic hydrocarbons (PAHs) compounds to sediments of the Niger Delta, Nigeria were assessed using combined chemical analysis and toxicity bioassay techniques. Concentrations of two- to six-ring PAHs of molecular mass 128-278 and toxicity to Vibrio fischeri and Lemna minor are considered in this investigation. Levels of the sum of the 16 USEPA priority pollutant PAHs varied from 20.7 to 72.1 ng/g dry weight. Sediment PAH levels were highest in samples collected from Delta Steel located at the outskirts of Warri, and Quality control centre, Ughelli West; with total PAH concentrations of 72.1 and 67.5 ng/g dry weight, respectively. The overall levels of PAHs in this study are low compared to other regions and reveal moderate PAHs pollution in the sediments of the Niger Delta. Two- and three-ring aromatic hydrocarbons predominated in almost all the sediments, which indicate a petrogenic origin. The sediment total PAH (PAHtot) concentration, normalized to organic carbon content (OC), ranged from 120.2 to 1.99 ng PAHtot/mg OC; and showed distinctively that the sedimentary organic matter of the sample from Delta Steel is highly contaminated with PAHs, and had a value of 120.2 ng PAHtot/mg OC. The toxicity bioassays indicated that the sample collected from Warri Refinery Area (SDWRR) was the most toxic to V. fischeri, with an EC50 value of 0.45 mg sediment equiv./mL test medium; and samples from Ogunu (SDOGN) and Warri Refinery area (SDWRR) showed high toxicity to L. minor, with percent inhibitions of 42.6% and 33.67%, respectively, after 7 days of exposure. The total PAH concentrations showed no correlation with toxicity bioassays, and thereby implied that chemical analysis of PAHs cannot be an indicator of sediment toxicity.

Aliivibrio fischeri↗

PubMed2005

Identification of toxicants from marine sediment using effect-directed analysis.

Effect-directed analysis (EDA) has been reported to be a powerful tool for the identification of the responsible toxicants in contaminated, hazardous environmental samples. The aim of this study was to investigate whether it also is possible to use currently available EDA methodology to identify potentially relevant toxicants in samples that do not pose obvious problems. For this purpose, compounds extracted from a marine sediment sample from the west coast of Sweden were separated into distinct fractions, using two preparative chromatographic techniques. One algal bioassay using Scenedesmus vacuolatus and two bacterial bioassays using Vibrio fischeri were applied as detectors of toxicity, representing acute and chronic end points. Chronic algal toxicity was a powerful tool for discriminating between toxic and nontoxic fractions, whereas acute and chronic bacterial toxicity failed to identify toxic fractions. Eight compounds were identified as potentially relevant toxicants by chemical analysis of toxic fractions: anthracene, fluoranthene, pyrene, benzo[a]anthracene, benzo[b]fluoranthene, benzo[a]pyrene, benzo[k]fluoranthene, and indeno[1,2,3-cd]pyrene.

Aliivibrio fischeri↗

PubMed2005

Structural alerts--a new classification model to discriminate excess toxicity from narcotic effect levels of organic compounds in the acute daphnid assay.

Quantitative and qualitative structure-activity relationships (QSARs) have a great potential to support the risk assessment of chemicals, provided there are tools available that allow evaluation of the suitability of QSARs for the compounds of interest. In this context, a pragmatic approach is to discriminate excess toxicity from narcotic effect levels, because the latter can be estimated from QSARs and thus have a low priority for experimental testing. To develop a respective scheme for the acute daphnid toxicity as one of the primary ecotoxicological endpoints, 1067 acute toxicity data entries for 380 chemicals involving the daphnid species Daphnia magna were taken from the on-line literature, and quality checks such as water solubility were employed to eliminate apparently odd data entries. For 36 known narcotics with LC50 values referring to D. magna, a reference baseline QSAR is derived. Compounds with LC50 values above a certain threshold defined relative to their predicted baseline toxicity are classified as exerting excess toxicity. Three simple discrimination schemes are presented that enable the identification of excess toxicity from structural alerts based on the presence or absence of certain heteroatoms and their chemical functionality. Moreover, a two-step classification approach is introduced that enables a prioritization of organic compounds with respect to their need for experimental testing. The discussion includes reaction mechanisms that may explain the association of structural alerts with excess toxicity, a comparison with predictions derived from mode of action-based classification schemes, and a statistical analysis of the discrimination performance in terms of detailed contingency table statistics.

Animal Testing Alternatives↗

PubMed2005

Predicting and observing responses of algal communities to photosystem II-herbicide exposure using pollution-induced community tolerance and species-sensitivity distributions.

Various test strategies are in use in ecotoxicology to assess the potential risks of toxicants on aquatic communities. The species-sensitivity distribution concept (SSD) works by arranging single-species laboratory test data in a cumulative frequency distribution. The pollution-induced community tolerance concept (PICT) uses observable community responses by measuring increases in community tolerance caused by the replacement of sensitive species after exposure. The aim of this study was to compare these two concepts in assessing the effects of three herbicides. Atrazine, prometryn, and isoproturon were found to increase community tolerance by a factor up to six. Atrazine increased community tolerance only at higher test concentrations (0.125 mg L(-1)). Species-sensitivity distributions correspond well to community responses: The median effective concentrations (EC50s) of untreated periphyton communities tested covered 55 to 65% of affected species represented in the SSD. The sensitivities of tolerant algal communities shifted to the right end of the SSDs. In the microcosm experiments, higher test concentrations affected biomass, species numbers, and community structure. Community tolerance could not be induced any further, suggesting that these concentrations represent a maximum of functional redundancy of a functional group. At higher concentrations, even the least-sensitive species are affected. These results can be interpreted as a confirmation of the SSD concept by observed algal community responses, when applied to photosystem II (PSII)-inhibiting herbicides.

Atrazine↗

PubMed2005

Algal toxicity of nitrobenzenes: combined effect analysis as a pharmacological probe for similar modes of interaction.

An analysis regarding the effects of the mixture toxicity of different nitrobenzenes on the reproduction of the green alga Scenedesmus vacuolatus was undertaken using the concepts of concentration addition and response addition. Using lipophilicity-based quantitative structure-activity relationship (QSAR) modeling for nitrobenzenes, the assumption is held that mononitrobenzenes may exert narcotic effects as a common type of action, whereas dinitrobenzenes show a somewhat greater toxicity. From the literature, QSARs based on quantum chemical parameters suggest that some mononitrobenzenes may be effective through additional other modes of action. The toxicity of a mixture of 14 nitrobenzenes clearly exceeds the predicted combined effects, as expected for the sum of toxic units from a uniform narcotic mode of action. Moreover, the observed combined effect is smaller than that predicted from similarly acting compounds calculated on the basis of the parameterized dose-response functions using concentration addition. Further modeling of the combined effect, joining the models of concentration addition for components with anticipated similar modes of action and of response addition for those with independent action, led us to propose that not all nitrobenzenes follow the same mode of action. This idea is in line with the hypothesis derived from quantum chemical QSAR considerations. Most interestingly, the methodology introduced here uses combined effect analysis as a pharmacological probe to test for similarity in the mode of action of mixture components.

Animals↗

PubMed2005

Confirmation of cause-effect relationships using effect-directed analysis for complex environmental samples.

The establishment of causal links between chemical contamination and observed ecotoxic effects of environmental samples is a major challenge in ecotoxicology. Effect-directed analysis is an approach combining fractionation with toxicity testing before chemical nontarget analysis to separate and subsequently identify toxic compounds in environmental samples. A final confirmation step in this procedure evaluates how much of the observed toxicity in the environmental sample can be attributed to the mixture of identified toxicants. The aim of the present study is to advance the methodology for the confirmation of mixtures in effect-directed analysis by using tools for the assessment of mixture toxicity accounting for unknown modes of action and heterogeneity of concentration-response curves. For this purpose, toxicants were identified in sediment extracts from two different sites. All identified compounds were tested both individually as pure compounds and in mixtures at ratios equal to those found in the sediment extracts. The observed extract toxicity was then compared with the expected combined effects calculated according to the models of concentration addition and independent action as well as with the observed toxicity of the synthetic mixture. Drawbacks of the established approach using toxic unit summation are demonstrated, and the Index of Confirmation Quality, an easy-to-read representation that allows a quantitative measure of confirmation over a range of different effect levels, is introduced.

Chemical Fractionation↗

PubMed2005

Effects of hydrogen sulfide to Vibrio fischeri, Scenedesmus vacuolatus, and Daphnia magna.

The effects of hydrogen sulfide (H2S) were tested in three ecotoxicological tests in order to evaluate its confounding potential in assessment of pore water and groundwater toxicity. The luminescent bacteria Vibrio fischeri, the water flea Daphnia magna, and the microalgae Scenedesmus vacuolatus often are part of a biotest battery. A new technique for the synthesis of hydrogen sulfide solutions of defined concentrations using an electrochemical generator instead of sodium sulfide solutions was used. Because hydrogen sulfide is volatile, the loss rate of H2S was studied over time to enable estimation of the mean test concentrations over the whole test duration. Loss rates were calculated to be 13 +/- 6% after 30 min, and 39 +/- 11% and 43 +/- 16% after a 24- and 48-h exposure time, respectively. Sensitivities of the test organisms in terms of median effective concentration (EC50), corrected for the above loss rates, varied from 0.28 to 0.0036 and 0.055 mM for the luminescent bacteria, the crustacea, and the algae, respectively. A species-sensitivity distribution using EC and mean lethal concentration literature data for marine and freshwater crustaceans and phytoplankton showed a medium sensitivity of the water flea D. magna, though the bacteria V. fischeri and the algae S. vacuolatus were among the least-sensitive group of organisms. This demonstrates that only the algae and the bacteria are easy to use in the assessment of toxicity of matrices with H2S concentrations above 0.06 mM.

Air Pollutants↗

PubMed2005

Light as a confounding factor for toxicity assessment of complex contaminated sediments.

Polycyclic aromatic hydrocarbons (PAHs) are contaminants often found in sediments. Their relevant contribution to toxic effects induced by environmental samples has been demonstrated using effect-directed analysis (EDA). Toxicity of PAHs previously has been reported to depend on light conditions. The aim of this study was to analyze the influence of simulated sunlight, in comparison to standard algal growth light, on the toxicity of samples, in which PAHs were identified as major toxicants using EDA. Additionally, toxicity of identified toxicants and mixtures of these compounds were assessed. It can be shown that the samples, the PAH compounds, and the created mixtures exhibit photoenhanced toxicity. The combined effects of the mixtures can be predicted using the model of concentration addition. This is surprising because different modes of action have been reported to contribute to phototoxicity of the identified toxicants. For the confirmation step in EDA, the toxicity assessment under simulated sunlight reveals that less of the samples' toxicity can be explained by the identified compounds, compared to the assessment conducted under standard growth light. Finally, the relevance of light conditions in the toxicity assessment is demonstrated for eight out of 13 transect samples of sediment extracts from river Elbe, Germany.

Confounding Factors, Epidemiologic↗

PubMed2004

What contributes to the combined effect of a complex mixture?

The effect of a mixture of 10 compounds, which have previously been identified in an effect-directed analysis as potentially relevant for a specific contaminated riverine sediment (Brack et al. Arch. Environ. Contam. Toxicol. 1999, 37, 164), were investigated for the underlying joint effect. Components identified in an organic sediment extract included several PAHs (benzo[ghi]fluoranthene, benz[a]anthracene, fluoranthene, pyrene, 2-phenylnaphthalene, anthracene, and phenanthrene) plus prometryn, N-phenyl-2-naphthylamine, and parathion-methyl. Experiments were performed using a one-generation algal bioassay with the unicellular green algae Scenedesmus vacuolatus as well as chlorophyll fluorescence quenching analysis to describe the effects of the components and mixtures thereof. Analysis of the mixture effects based on concentration-response modeling of the effect data reveals that indeed effect contributions of several components can be expected although the mixture ratio is not equitoxic and the individual components vary greatly with respect to biological effect. Comparing predicted and observed mixture effects, the combined effect may not be attributed to a joint narcotic effect of the mixture components. Evidently, some of the components act specifically and dissimilar and may therefore be best described in their combined effect by response addition while for others a similar mode of action seems plausible. Chlorophyll fluorescence quenching analysis supports to discriminate between prometryn, N-phenyl-2-naphthylamine, and PAHs. A joint model for calculating the combined effect using concentration addition for the suspected unspecifically acting components in algae (PAHs and parathion-methyl) and subsequently response addition for this group and the other components clearly improves the description of the observed combined effect. Allocation of effect contributions to specific components using toxic units or effect contributions lead to different judgments. The observed combined effect of a 3-compound mixture of prometryn, N-phenyl-2-naphthylamine, and benzo[ghi]fluoranthene is indistinguishable from the effects of the original 10-compound mixture, demonstrating the need in site-specific assessment of complex contamination to account for the mode of action of contaminants. Implications forthe confirmation step in effect-directed analysis of substances causing effects in complex contaminated samples are discussed.

2-Naphthylamine↗

PubMed2004

On line biomonitors used as a tool for toxicity reduction evaluation of in situ groundwater remediation techniques.

Success of groundwater remediation is typically controlled via snapshot analysis of selected chemical substances or physical parameters. Biological parameters, i.e. ecotoxicological assays, are rarely employed. Hence the aim of the study was to develop a bioassay tool, which allows an on line monitoring of contaminated groundwater, as well as a toxicity reduction evaluation (TRE) of different remediation techniques in parallel and may furthermore be used as an additional tool for process control to supervise remediation techniques in a real time mode. Parallel testing of groundwater remediation techniques was accomplished for short and long time periods, by using the energy dependent luminescence of the bacterium Vibrio fischeri as biological monitoring parameter. One data point every hour for each remediation technique was generated by an automated biomonitor. The bacteria proved to be highly sensitive to the contaminated groundwater and the biomonitor showed a long standing time despite the highly corrosive groundwater present in Bitterfeld, Germany. The bacterial biomonitor is demonstrated to be a valuable tool for remediation success evaluation. Dose response relationships were generated for the six quantitatively dominant groundwater contaminants (2-chlortoluene, 1,2- and 1,4-dichlorobenzene, monochlorobenzene, ethylenbenzene and benzene). The concentrations of individual volatile organic chemicals (VOCs) could not explain the observed effects in the bacteria. An expected mixture toxicity was calculated for the six components using the concept of concentration addition. The calculated EC(50) for the mixture was still one order of magnitude lower than the observed EC(50) of the actual groundwater. The results pointed out that chemical analysis of the six most quantitative substances alone was not able to explain the effects observed with the bacteria. Thus chemical analysis alone may not be an adequate tool for remediation success evaluation in terms of toxicity reduction.

Aliivibrio fischeri↗

PubMed2003

Mixture toxicity and its modeling by quantitative structure-activity relationships.

Environmental contaminants are frequently encountered as mixtures, and the behavior of chemicals in a mixture may not correspond to that predicted from data on the pure compounds. This paper reviews current quantitative structure-activity relationship (QSAR) methodology for the analysis of mixture toxicity. Interactions of components in a mixture can cause complex and substantial changes in the apparent properties of its constituents, resulting in synergistic or antagonistic effects as opposed to the ideal reference case of additive behavior: concentration addition (CA) and independent action (IA) are two prominent reference models for the evaluation of joint activity, and both have mechanistic support from pharmacology. After discussing graphical tools for analyzing binary mixtures and joint effect indices suitable also for multicomponent mixtures, water solubility and hydrophobicity of mixtures are analyzed with respect to the property contributions of the individual components. With the former, small but significant deviations from ideal behavior are observed even for simple organics, whereas in the case of low concentrations, mixture hydrophobicity was found to agree approximately with the fractional contributions of the components. A variety of studies suggest that mixtures of compounds exerting only one (narcotic or specific) mode of action can be modeled satisfactorily by assuming CA, whereas the interaction of differently acting compounds tends to yield a less than CA joint activity. The QSARs have been used to predict concentrations of components in mixtures from joint effects and defined mixture ratios and have been developed to predict narcotic-type mixture toxicity from molecular descriptors that are calculated as composite properties according to the fractional concentrations of the mixture components. In the case of ionogenic compounds, initial results suggest that CA may serve as a first-order approximation for the joint effect of un-ionized and ionized compound portions.

Animals↗

PubMed2003

Identification of toxic products of anthracene photomodification in simulated sunlight.

Currently, the evidence of a rapid photomodification of anthracene under sunlight resulting in enhanced toxicity exists; however, the chemical causes of toxicity are still unknown. The present study aimed at filling this gap by irradiation of an anthracene suspension with simulated sunlight and subsequent effect-directed fractionation and analysis of toxic products with respect to the inhibition of bacterial energy metabolism of Vibrio fischeri, reproduction of the green algae Scenedesmus vacuolatus, and genotoxicity in the umuC test. Algal toxicity of anthracene was hardly modified by irradiation prior to testing and distributed over all fractions with emphasis on the fractions containing anthracene-9,10-dione and a photometabolite suggested to be 10-hydroxyanthrone. Bacterial toxicity and genotoxicity in contrast emerged only when anthracene was irradiated. Anthracene-1,4-dione, a so-far-unknown trace photometabolite, was identified as a very potent toxicant dominating the toxicity of photomodified anthracene to V. fischeri. In genotoxic fractions, 1-hydroxyanthracene-9,10-dione and 1,4-dihydroxyanthracene-9,10-dione were identified and confirmed as genotoxicants. The results stress the potential of effect-directed analysis approaches in contrast to mere chemical analysis in studies aiming at toxicologically relevant photomodified substances.

Anthracenes↗

PubMed2002

Mixture toxicity of priority pollutants at no observed effect concentrations (NOECs).

Environmental exposure situations are often characterised by a multitude of heterogeneous chemicals with ambiguous or unknown modes of action present at low concentrations. While multiple exposure is widely acknowledged, arguments are raised that adverse combined effects might not be evoked by mixtures of substances with dissimilar modes of action and being present at only low concentrations. In this study the combined effect of a multiple mixture composed of structurally dissimilar priority pollutants with mostly unknown modes of action has been investigated using an algal biotest. The concentrations of the components in the mixture equalled statistically estimated, individual no observed effect concentrations (NOECs). The observed mixture toxicity was not only clearly higher than expected for any single substance alone, but also well predictable using the concept of independent action.

Atrazine↗

PubMedDate not supplied

Combination effect of light and toxicity in algal tests.

The sensitivity of Scenedesmus subspicatus against potassium dichromate is positively correlated to the photon flux density during the algal growth inhibition test. Low photon flux densities led to significantly reduced maximum effects and higher EC50 levels. To improve the testing of colored substances, we distinguished between the toxic effect (chemical part, represented by potassium dichromate) and the shading effect (physical part, simulated by reduced light intensities during the test) of a hypothetical light absorbing substance. The contribution of these single effects to the total inhibition varied greatly. At high concentrations of potassium dichromate (1.6 and 3.2 mg L(-1)) the physical part never exceeded 25% of the total inhibition, not even at strongest light reduction, while at low concentrations (0.2 and 0.4 mg L(-1)) the physical effect became more prominent when halving the amount of available light. Further, the combination effect of the chemical and the physical effect could be calculated well only by using the concept of independent action. Thus, if chemical and physical effects are measured in combination, as is the case in tests with dye-stuffs, the current test protocol for the algal growth inhibition test may lead to incorrect estimations of the toxic potential.

Chlorophyta↗