Glycosylation of bisphenol A by freshwater microalgae

The endocrine disruptor bisphenol A (BPA, 4,4'-isopropylidenediphenol) is used to manufacture polycarbonate plastic and epoxy resin linings of food and beverage cans, and the residues from these products are then sometimes discharged into rivers and lakes in waste leachates. However, the fate of BPA in the environment has not yet been thoroughly elucidated. Considering the effect of BPA on aquatic organisms, it is important that we estimate the concentration of BPA and its metabolites in the aquatic environment, but there are few data on the metabolites of BPA. Here, we focused on freshwater microalgae as organisms that contribute to the biodegradation or biotransformation of BPA in aquatic environments. When we added BPA to cultures of eight species of freshwater microalgae, a reduction in the concentration of BPA in the culture medium was observed in all cultures. BPA was metabolized to BPA glycosides by Pseudokirchneriella subcapitata, Scenedesmus acutus, Scenedesmus quadricauda, and Coelastrum reticulatum, and these metabolites were then released into the culture medium. The metabolite from P. subcapitata, S. acutus, and C. reticulatum was identified by FAB-MS and (1)H-NMR as bisphenol A-mono-O-beta-d-glucopyranoside (BPAGlc), and another metabolite, from S. quadricauda, was identified as bisphenol A-mono-O-beta-d-galactopyranoside (BPAGal). These results demonstrate that freshwater microalgae that inhabit universal environments can metabolize BPA to its glycosides. Because BPA glycosides accumulate in plants and algae, and may be digested to BPA by beta-glycosidase in animal intestines, more attention should be given to levels of BPA glycosides in the environment to estimate the ecological impact of discharged BPA.     

Cyst-based toxicity tests. XI. Influence of the type of food on the intrinsic growth rate of the rotifer Brachionus calyciflorus in short-chronic toxicity tests

As important members of zooplankton communities worldwide, rotifers are used extensively in ecotoxicological research. Chronic rotifer tests are, however, dependent on live algal food which adds to the complexity, the variability and the costs of these bioassays. To bypass the former problem, experiments have been undertaken with the freshwater rotifer Brachionus calyciflorus, to determine their intrinsic growth rate (r) when fed for 48 h on a mixture of green algae (Raphidocelis subcapitata recently renamed Pseudokirchneriella subcapitata) obtained from algal beads stored for different periods of time, and other inert foods. All tests have been performed in disposable multiwells, in 1 ml cups each inoculated with 1 rotifer freshly hatched from dried cysts. The majority of the growth tests was performed in eight replicates. The investigations revealed that microalgae from algal beads stored for up to one year, in darkness, at 4 degrees C, supplemented with dried blue-green algae (Spirulina) gave satisfactory rotifer reproduction. The intrinsic growth rates of the rotifers, were, however, dependent on the storage time of the algal beads; the highests r's (0.7-0.8) were obtained with algae from beads not older than four months. Growth tests with combinations of P. subcapitata and other inert feeds revealed that the enrichment food Selco used in aquaculture, also gave the same reproductive output as the combination microalgae/Spirulina. A rotifer growth experiment with 18 replicates showed that the variation coefficient is below 20% when the tests comprise eight replicates. This study demonstrated that microalgae from beads, supplemented by other inert food, open the door for a practical and cost-effective short-chronic rotifer test, which is totally independent of the culturing of both the test species and its live food.     

赤泥复合剂絮凝处理太湖水华藻与海洋赤潮藻

研究了一种新型赤泥复合剂对太湖夏季水华原水、实验室培养海洋赤潮藻及模拟海水的絮凝除藻降浊性能。结果表明,赤泥复合剂具有良好的除藻降浊效果,对太湖水华藻去除率为97.3%,原水浊度去除率为95%;对海洋小球藻和三角褐指藻2种赤潮藻去除率分别高达99.1%和98.9%;对2种模拟海水的最高除浊率分别为96.3%和95.3%,其剩余浊度均低于4 NTU,从而为治理淡水水华与海洋赤潮提供一种新技术,同时为赤泥综合利用提供一种新途径。     

Comparative toxicity of acrylic acid to marine and freshwater microalgae and the significance for environmental effects assessments.

In this study, we compared the sensitivity of freshwater and marine organisms to two structurally similar substances, acrylic acid and methacrylic acid. Reported acute toxicity data (L(E)C50-values) for freshwater organisms range from 0.1 to 222 mg/l and 85 to >130 mg/l for acrylic acid and methacrylic acid, respectively. The large variation in toxicity data for acrylic acid is due to a specific toxicity to certain species of freshwater microalgae, with algae EC50-values being two to three orders of magnitude lower than L(E)C50-values reported for fish and invertebrates. To evaluate the sensitivity of marine organisms, ecotoxicity data was generated for ten species of microalgae, one invertebrate species and one fish species. For methacrylic acid, we found a marine acute toxicity that ranged from 110 to >1260 mg/l, which is comparable to reported data on freshwater organisms. In strong contrast, the resulting L(E)C50-values for acrylic acid ranged from 50 to >1000 mg/l, and there was no specific sensitivity of marine algae when compared to marine invertebrates and fish. For acrylic acid, therefore, use of the available freshwater toxicity data for an effects assessment for the marine environment is likely to overestimate the hazard and risk from this substance. Overall, the results of the study suggest that ecotoxicity data generated on freshwater species may not always be appropriate for the effects assessments of organic chemicals in the marine environment, thus emphasising the importance of using ecologically relevant data to assess environmental risk.     

Effects of bensulfuron-methyl and cinosulfuron on growth of four freshwater species of phytoplankton

The acute toxicity of sulfonylurea herbicides bensulfuron-methyl and cinosulfuron was tested on the five species of freshwater phytoplankton: Scenedesmus acutus, Scenedesmus subspicatus, Chlorella vulgaris and Chlorella saccharophila. Herbicide concentrations eliciting a 50% growth reduction over 96 h (EC50) ranged from 8 to 104 mg/l for cinosulfuron and from 0.015 to 6.2 mg/l for bensulfuron-methyl. The pesticides bensulfuron-methyl, atrazine and benthiocarb were more toxic than cinosulfuron, chlorsulfuron, molinate, fenitrothion and pyridaphenthion in a toxicity study with the same algal species. The transformation of effective concentrations of bensulfuron-methyl and cinosulfuron and other pesticides, obtained from toxicity measurements, into percent of the saturation level in water is used as a first evaluation of potential hazard to aquatic systems. The herbicides cinosulfuron, methyl-bensulfuron, atrazine and chlorsulfuron were more dangerous than the herbicides benthiocarb and molinate and than the insecticides fenitrothion and pyridaphenthion, in a study of hazard evaluation. The two species of Chlorella were more tolerant to both herbicides than the two species of Scenedesmus. A potential environmental hazard of sulfonylurea herbicides to aquatic systems has to be expected even at low environmental concentrations.     

Acute toxicity and genotoxicity of five selected anionic and nonionic surfactants

The results of four toxicity bioassays of selected anionic and nonionic surface active agents were presented. Three widely used anionic surfactants that belong to alkyl sulphates (AS), alkylbenzene sulphonates (LAS) and alkylpolyoxyethylene sulphates (AES) as well as nonionic surfactants: polyoxyethylene alkyl ethers (AE) and polyoxylethylene alkylphenyl ethers (APE) were tested. Three different toxicity assays to aquatic organisms: Physa acuta Draparnaud, Artemia salina and Raphidocelis subcapitata were applied. Additionally, the genotoxicity test with Bacillus subtilis M45 Rec- and H17 Rec+ strains was performed. The obtained results showed that none of the surfactants studied was genotoxic at the concentration 1000 mg l(-1). On the basis of toxicity tests to aquatic organisms all tested anionic surfactants were harmful (LC50 between 10 and 100 mg l(-1)), whereas nonionic ones were toxic (LC50 between 1 and 10 mg l(-1)) or even highly toxic (LC50 below 1 mg l(-1)). Moreover, the bigger was the molecular weight of the tested compound, the higher toxicity was observed.     

Biological variation in sensitivity to N-heterocyclic PAHs; effects of acridine on seven species of micro-algae

The toxicity of the nitrogen (N) heterocyclic polyaromatic hydrocarbon (PAH) acridine was tested for seven species of microalgae: Scenedesmus acuminatus, Selenastrum capricornutum, Chlamydomonas eugametos, Staurastrum chaetoceras, Staurastrum manfeldtii, Navicula salinarum and Nitzschia sigma. The effect of acridine on the algae was studied in a 96-h growth test, in which growth rates were determined using cell numbers and biovolume. The obtained EC50 values (for growth rates based on cell numbers) ranged from 0.08 mg litre(-1) for N. sigma to 0.78 mg litre(-1) for C. eugametos and N. salinarum. Effect concentrations based on biovolume were slightly higher for most species. Metabolism of acridine was observed for one species (S. capricornutum), but this capacity did not result in a very different tolerance. Acridine toxicity was neither related to taxonomical background (green algae versus diatoms) nor to original habitat of the species (planktonic or benthic, eutrophic or oligo-mesotrophic). The presence of near-UV radiation during the incubation might explain the higher toxicity of acridine than is expected on basis of QSAR derived narcotic toxicity.     

Photo-oxidation of Sb(III) in the seawater by marine phytoplankton-transition metals-light system

The photo-oxidation of Sb(III) to Sb(V) by marine microalgae (diatom, green and red algae) with or without the presence of transition metals (Fe(III), Cu(II) and Mn(II)). The influence of marine phytoplankton on the photochemistry of antimony was confirmed for the first time. The conversion ratio of Sb(III) to Sb(V) increased with increasing algae concentration and irradiation time. Different species of marine phytoplankton were found to have different photo-oxidizing abilities. The photochemical redox of transition metals could induce the species transformation of antimony. After photo-induced oxidation by marine phytoplankton and transition metals, the ratio of Sb(V) to Sb(III) was in the range of 1.07-5.48 for six algae (Tetraselmis levis, Chlorella autotrophica, Nannochloropsis sp., Tetraselmis subcordiformis, Phaeodactylum tricornutum, and Porphyridium purpureum), and only 0.92 for Dunaliella salina. The distribution of antimony in the sunlit surface seawater was greatly affected by combined effects of marine phytoplankton (main contributor) and transition metals; both synergistic and antagonistic effects were observed. The results provided further insights into the distribution of Sb(III) and Sb(V) and the biogeochemical cycle of antimony, and have significant implications for the risk assessment of antimony in the sunlit surface seawater.     

Toxicity of methyl tert-butyl ether to marine organisms: ambient water quality criteria calculation

In response to increasing concerns over the detection of methyl tert-butyl ether (MTBE) in groundwater and surface water and its potential effects in aquatic ecosystems, industry and the United States Environmental Protection Agency (USEPA) began to collaborate in 1997 to develop aquatic toxicity databases sufficient to derive ambient water quality criteria for MTBE consistent with USEPA requirements. Acute toxicity data for seven marine species, chronic toxicity data for an invertebrate, and plant toxicity data were developed to complete the saltwater database. The species tested were Cyprinodon variegatus, Gasterosteus aculeatus, Callinectes sapidus, Mytilus galloprovincialis, Palaemonetes pugio, Rhepoxynius abronius, Americamysis bahia, and Skeletonema costatum. The toxicity tests were conducted in accordance with USEPA and American Society for Testing and Materials testing procedures and Good Laboratory Practice guidelines. Data developed from this study were consistent with existing data and showed that MTBE has low acute and chronic toxicity to the marine species tested. Based upon measured MTBE concentrations, acute effects were found to range from 166 mg MTBE/l for the grass shrimp to 1950 mg MTBE/l for marine mussel. The no-observed effect concentration for the reproduction and growth of mysids was 26 mg MTBE/l during the life cycle test. The toxicity of MTBE to saltwater organisms is comparable to its toxicity to the freshwater species tested. Reported MTBE concentrations in coastal waters are several orders of magnitude lower than concentrations observed to cause effects in marine organisms.     

Toxicities of 48 pharmaceuticals and their freshwater and marine environmental assessment in northwestern France

A risk assessment for freshwater and marine ecosystems is presented for 48 pharmaceutical compounds, belonging to 16 therapeutic classes, and prescribed in northwestern France. Ecotoxicity data were obtained on two freshwater organisms, i.e., crustacean Daphnia magna and the green algae Pseudokirchneriella subcapitata, and on two marine organisms, i.e., the crustacean Artemia salina and the diatom Skeletonema marinoi. Measured environmental concentrations (MEC), in the Orne River and sea off Merville-Franceville in the Basse-Normandie region, were compared to the predicted environmental concentrations (PEC). Predicted no-effect concentrations (PNEC) were derived from acute data for each compound. Then, a risk assessment for each compound and the mixture was performed by calculating risk quotients (RQ as PEC or MEC/PNEC ratio). Results showed that no immediate acute toxicities were expected even if some compounds displayed strong toxicities at very low concentrations. Antibiotics, antidepressants, and antifungals would deserve attention because of their high or median ecological risk suspected on marine and freshwater ecosystems. Marine ecosystems would be more sensitive to pharmaceutical residues.     

Toxicity of fluoranthene and its biodegradation metabolites to aquatic organisms

The toxicity of nine stable products of the biodegradation of fluoranthene with the pure bacterial strain Pasteurella sp. IFA was studied. For their quantification, an improved analytical procedure with two-step liquid-liquid extraction, derivatisation and gas chromatographic-mass spectrometric detection was used. Growth inhibition and immobility tests for fluoranthene and its metabolites were carried out using algae (Scenedesmus subspicatus), bacteria (Pseudomonas putida) and crustaceans (Daphnia magna and Thamnocephalus platyurus). Tests using the alga S. subspicatus revealed that with the exception of 9-hydroxyfluorene, which was only four times less toxic than fluoranthene, all the other metabolites were 37 to approximately 3000 times less toxic than the parent material. P. putida cells were resistant to fluoranthene and its primary metabolites, but were inhibited by low molecular weight intermediates, especially benzoic acid. Fluoranthene was not toxic to T. Platyurus, but was toxic to D. magna. Its primary metabolites (including 9-fluorenone and 9-hydroxyfluorene) were toxic to D. magna, and a low molecular weight metabolite (2-carboxybenzaldehyde) was highly toxic to T. platyurus.     

Growth assays with mixed cultures of cyanobacteria and algae assessed by in vivo fluorescence: One step closer to real ecosystems?

A growth toxicity assay with mixed cultures of cyanobacteria and algae using in vivo fluorescence is presented. Test organisms (the green alga Pseudokirchneriella subcapitata and the cyanobacterium Aphanothece clathrata) growing alone and in a mixture were exposed to selected chemicals. P. subcapitata featured a higher sensitivity to toxicants in the presence of A. clathrata compared to the single species assay. On the other hand, growth of a cyanobacterium was not affected by the presence or absence of the green alga. The proposed method seems to be suitable for pre-screening studies of toxicants (algistatic agents, herbicides) applied into the aquatic environment and for the assessment of their impact on natural phytoplankton communities.     

Effects of 2,4-D, glyphosate and paraquat on growth, photosynthesis and chlorophyll-a synthesis of Scenedesmus quadricauda Berb 614

The effects of 2,4-D, glyphosate and paraquat on growth, photosynthesis and chlorophyll-a synthesis by a freshwater green alga, Scenedesmus quadricauda Berb 614, were determined. These herbicides are the most often used in Hong Kong. Within the concentration range 0.02-200 mg/l, paraquat was more toxic than glyphosate and 2,4-D to the growth, photosynthesis and chlorophyll-a synthesis. The presence of 0.02, 0.2 or 2 mg/l of 2,4-D was not toxic to the alga. Algal growth, photosynthesis and chlorophyll-a synthesis were stimulated by the presence of low concentrations (0.02 or 0.2 and 0.02 mg/l, respectively) of 2,4-D and glyphosate. The presence of 0.02 or 0.2 mg/l of paraquat, 2 mg/l of glyphosate or 20 mg/l of 2,4-D was significantly inhibitory to the three test parameters, whereas the presence of 2 or more mg/l of paraquat, 20 or more mg/l of glyphosate or 200 mg/l of 2,4-D completely inhibited algal growth, photosynthesis and chlorophyll-a synthesis. The use of the alga as a bio-indicator of herbicide contamination in freshwater environment was discussed.     

Algal tests with soil suspensions and elutriates: a comparative evaluation for PAH-contaminated soils

An algal growth inhibition test procedure with soil suspensions is proposed and evaluated for PAH-contaminated soil. The growth rate reduction of the standard freshwater green alga Pseudokirchneriella subcapitata (formerly known as Selenastrum capricornutum) was used as the toxicity endpoint, and was quantified by measuring the fluorescence of solvent-extracted algal pigments. No growth rate reduction was detected for soil contents up to 20 g/l testing five non-contaminated Danish soils. Comparative testing with PAH-contaminated soil elutriates and soil suspensions showed that the suspensions had toxicity endpoints 2.5-3000 times lower than tests with the corresponding elutriates. Algal growth inhibition tests with soil suspensions are recommended for screening purposes as a supplement to elutriate testing. Experiments with a phenanthrene-spiked soil, showed that the sorbed compound did not contribute to the toxicity. However, the soil did act as a reservoir for phenanthrene, allowing desorption to occur continuously during the algal test which maintained higher concentrations of phenanthrene in the dissolved phase. Phenanthrene-spiked soil incubated for 90 days before algal testing, resulted in a reduction of the toxicity to P. subcapitata by a factor of 76 (from EC10 = 0.3 to 23.6 g soil/l). However, during this 90-day period the total concentration of phenanthrene in the soil decreased by 38% (from 322 to 199 mg/kg) indicating that phenanthrene in the aged soil had become less bioavailable.     

Preliminary ecotoxicological characterization of a new energetic substance, CL-20

A new energetic substance hexanitrohexaazaisowurtzitane (or CL-20) was tested for its toxicities to various ecological receptors. CL-20 (epsilon-polymorph) was amended to soil or deionized water to construct concentration gradients. Results of Microtox (15-min contact) and 96-h algae growth inhibition tests indicate that CL-20 showed no adverse effects on the bioluminescence of marine bacteria Vibrio fischeri and the cell density of freshwater green algae Selenastrum capricornutum respectively, up to its water solubility (ca. 3.6 mg l(-1)). CL-20 and its possible biotransformation products did not inhibit seed germination and early seedling (16-19 d) growth of alfalfa (Medicago sativa) and perennial ryegrass (Lolium perenne) up to 10,000 mg kg(-1) in a Sassafras sandy loam soil (SSL). Indigenous soil microorganisms in SSL and a garden soil were exposed to CL-20 for one or two weeks before dehydrogenase activity (DHA) or potential nitrification activity (PNA) were assayed. Results indicate that up to 10,000 mg kg(-1) soil of CL-20 had no statistically significant effects on microbial communities measured as DHA or on the ammonium oxidizing bacteria determined as PNA in both soils. Data indicates that CL-20 was not acutely toxic to the species or microbial communities tested and that further studies are required to address the potential long-term environmental impact of CL-20 and its possible degradation products.     

Effects of a surfactant (FFD-6) on Scenedesmus morphology and growth under different nutrient conditions

Surfactants are man-made compounds that are meanwhile omnipresent in the environment, but environmental concentrations of surfactants are such that they are thought to have little risk for aquatic systems. The major anionic surfactants currently on the global market are linear alkylbenzene sulfonates (LAS), a class where the commercially available FFD-6 belongs to. The hypothesis was tested that sublethal effects of FFD-6, i.e. the morphological effect of colony formation in the common test alga Scenedesmus obliquus, occurs at a concentration lower than the no-observed-effect concentrations for endpoints commonly used in regulatory toxicity testing with algae. The surfactant FFD-6 induced colonies in Scenedesmus at concentrations a few orders of magnitude lower (i.e. between 0.001 and 0.01 g l-1) than at which growth inhibition was observed (i.e. between 1 and 10g l -1). Growth rates were lowest for Scenedesmus grown in P-limited medium, intermediate for algae reared in N-limited medium and highest for algae cultured in non-limited standard medium. Growth inhibition due to FFD-6 was similar for non-limited and nutrient-limited Scenedesmus, but colony formation was stronger in non-limited Scenedesmus than in nutrient limited cultures. The colony inducing effect of the surfactant FFD-6 on Scenedesmus occurs at much lower concentrations than growth inhibition and might affect species interactions, the survival of species and the energy flow along the food chain.     

Comparative study on the susceptibility of freshwater species to copper-based pesticides

Copper compounds have been intentionally introduced into water bodies as aquatic plant herbicides, algicides and molluscicides. Copper-based fertilizers and fungicides have been widely used in agriculture as well. Despite the fact that copper is an essential element for all biota, elevated concentrations of this metal have been shown to affect a variety of aquatic organisms. Nonetheless, comparative studies on the susceptibility of different freshwater species to copper compounds have seldom been performed. This study was conducted to compare toxicity of copper-based pesticides (copper oxychloride, cuprous oxide and copper sulfate) to different freshwater target (Raphidocelis subcapitata, a planktonic alga and Biomphalaria glabrata, a snail) and non-target (Daphnia similis, a planktonic crustacean and Danio rerio, a fish) organisms. Test water parameters were as follows: pH = 7.4 +/- 0.1; hardness 44 +/- 1 mg/l as CaCO3; DO 8-9 mg/l at the beginning and > 4 mg/l at the end; temperature, fish and snails 25 +/- 1 degrees C, Daphnia 20 +/- 2 degrees C, algae 24 +/- 1 degrees C. D. similis (immobilization), 48-h EC50s (95% CLs) ranging from 0.013 (0.011-0.016) to 0.043 (0.033-0.057) mg Cu/l, and R. subcapitata (growth inhibition), 96-h IC50s from 0.071 (0.045-0.099) to 0.137 (0.090-0.174) mg Cu/l, were the most susceptible species. B. glabrata (lethality), 48-h LC50s from 0.179 (0.102-0.270) to 0.854 (0.553-1.457) mg Cu/l, and D. rerio (lethality), 48-h LC50s 0.063 (0.045-0.089), 0.192 (0.133-0.272) and 0.714 (0.494-1.016) mg Cu/l, were less susceptible than Daphnia to copper-based pesticides. Findings from the present study therefore suggest that increased levels of copper in water bodies is likely to adversely affect a variety of aquatic species.     

Effects of the parasiticide ivermectin on the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata

Although widely used for the treatment of endo- and ectoparasites in livestock and pets, very few data on chronic effects on aquatic organisms are available for the parasiticide ivermectin. In the present study, toxicity of ivermectin to two freshwater organisms, the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata was investigated. For D. magna, a mean LC(50) 48 h of 5.7 ngl(-1) was derived from 10 acute tests. Chronic toxicity of ivermectin to D. magna was extremely high: with 0.001 and 0.0003 ngl(-1), respectively, nominal LOEC and NOEC based on growth and reproduction were far below the analytical limit of detection for this compound. P. subcapitata was considerably less sensitive to ivermectin than D. magna. For both growth rate and yield, EC(50) was >4,000 microgl(-1), LOEC was 1,250 microgl(-1) and NOEC 391microgl(-1). In view of the high toxicity to D. magna, the use of ivermectin might pose a risk to local aquatic ecosystems. Further studies should be carried out to investigate the effects of ivermectin and its degradation products on pelagic and benthic freshwater invertebrates.     

Effects of propanil, tebufenozide and mefenacet on growth of four freshwater species of phytoplankton: a microplate bioassay

The Albufera Natural Park situated in Valencia (Spain), with a very rich flora and fauna is surrounded by rice fields in which pesticide spraying is a regular practice. With this in mind, the sensitivity of four algal species, Scenedesmus acutus, Scenedesmus subspicatus, Chlorella vulgaris and Chlorella saccharophila to pesticides propanil, tebufenozide and mefenacet was studied using single species toxicity tests. Organisms were exposed to different concentrations of these herbicides and the algal growth was measured in a microplate reader at 410 nm, at 0, 24, 48 and 72 h. Tebufenozide appeared to be the most inhibitory to Scenedesmus and Chlorella species growth. 72 h EC50 of propanil, tebufenozide and mefenacet ranged from 0.29 to 5.98 mg/l, 0.12 to 0.15 mg/l and from 0.25 to 0.67 mg/l, respectively for the four algal species. The two species of Chlorella were more tolerant than the two species of Scenedesmus.     

Toxicity of chlorophenols to Pseudokirchneriella subcapitata under air-tight test environment

A closed-system algal toxicity test with no headspace was applied to evaluate the toxicity of chlorophenols to Pseudokirchneriella subcapitata. The dissolved oxygen production and the growth rate based on cell density were the response endpoints. Phenol and seven chlorophenols were tested using the above test technique. Median effective concentrations (EC50) range from 0.004 to 25.93 mg/l (based on DO production) and 0.0134 to 20.90 mg/l (based on growth rate). No-observed-effect concentration (NOEC) is within the range of 0.001-8.19 mg/l. In general, growth rate is a more sensitive response endpoint than the oxygen production, except for the case of pentachlorophenol. However, the differences in sensitivity between the two parameters were marginal. Furthermore, quantitative structure-activity relationships (QSAR's) based on the n-octanol/water partition coefficient (log P) and the acid dissociation constant (pK(a)) values were established with R(2) ranged from 0.90 to 0.96. From literature data also based on P. subcapitata, the new test method is 1.65-108 times more sensitive than the conventional algal batch tests. A completely different relative-sensitivity relationship among various aquatic organisms was thus observed. The results of this study indicate that the toxicity data of volatile organic chemicals derived by conventional algal toxicity tests may severely underestimate the impact of these toxicants. Our results show that alga is very sensitive to chlorophenols compared to other aquatic organisms such as the luminescent bacteria (the Microtox test), Daphnia magna, and rainbow trout.