Cultivating Chlorella vulgaris and Scenedesmus quadricauda microalgae to degrade inorganic compounds and pesticides in water

Environmental Science and Pollution Research - This work evaluates the possibility of cultivating Scenedesmus quadricauda and Chlorella vulgaris microalgae in wastewater from the hydroponic...     

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.     

Performance of different microalgal species in removing nickel and zinc from industrial wastewater

A series of batch experiments was conducted to compare the ability of 11 microalgal species of the same cell density in removing nickel (Ni) and zinc (Zn) from synthetic wastewater. These included Chlorella vulgaris (commercially available), Chlorella sorokiniana and Scenedesmus quadricauda (isolates from polluted water of Wuhan, China), and eight different isolates from Hong Kong. The Wuhan isolate of Scenedesmus removed most Ni, probably due to its large biomass. Nickel concentration was reduced from an initial 30 to 0.9 mg/l after 5 min (97% Ni removal), and further declined to 0.4 mg/l after 90 min of treatment. In wastewater containing 30 mg/l Ni and 30 mg/l Zn, more than 98%, Ni and Zn were removed simultaneously at the end of 5 min treatment, indicating that the presence of Zn in wastewater did not affect Ni removal by this Scenedesmus isolate. The second most effective species for Ni removal was an isolate, tentatively identified as Chlorella miniata, Ni concentration was reduced to 10 mg/l after 90 min, and was only slightly interfered by the presence of Zn. In terms of metal removal per unit biomass or unit surface area of algal cells, C. miniata was the best species in removing Ni and Zn. At the other extreme, one Hong Kong isolate (Synechocystis sp.) did not remove any Ni and only achieved 40% Zn removal. Performance of the other isolates was comparable with the commercial C. vulgaris, less than 50% Ni was removed after 5 h of treatment and Ni removal was significantly reduced by the presence of Zn. All algae tested were found to be viable, showing these 11 species could tolerate a mixture of 30 mg/l Ni and 30 mg/l Zn in wastewater.     

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.     

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.     

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.     

Kinetics of inorganic carbon utilization by microalgal biofilm in a flat plate photoreactor

A kinetic model was developed to describe inorganic carbon utilization by microalgae biofilm in a flat plate photoreactor. The model incorporates the fundamental mechanisms of diffusive mass transport and biological reaction of inorganic carbon by microalgal biofilm. An advanced numerical technique, the orthogonal collocation method and Gear's method, was employed to solve this kinetic model. The model solutions included the concentration profiles of inorganic carbon in the microalgal biofilm, the growths of suspended microalgae and microalgal biofilm, the effluent concentrations of inorganic carbon, and the flux of inorganic carbon from bulk liquid into biofilm. The batch kinetic test was independently conducted to determine biokinetic parameters used in the microalgal biofilm model simulation while initial thickness of microalgal biofilm were assumed. A laboratory-scale flat plate photoreactor with a high recycle flow rate was setup and conducted to verify the model. The volume of photoreactor is 60 l which yields a hydraulic retention time of 1.67 days. The model-generated inorganic carbon and the suspended microalgae concentration curves agreed well with those obtained in the laboratory-scale test. The fixation efficiencies of HCO(3)(-) and CO(2) are 98.5% and 90% at a steady-state condition, respectively. The concentration of suspended microalgal cell reached up to 12 mg/l at a maximum growth rate while the thickness of microalgal biofilm was estimated to be 104 microm at a steady-state condition. The approaches of experiments and model simulation presented in this study could be employed for the design of a flat plate photoreactor to treat CO(2) by microalgal biofilm in a fossil-fuel power plant.     

Biodegradation capacity of tributyltin by two Chlorella species

Two microalgal species, Chlorella vulgaris and Chlorella sp., which showed high tributyltin (TBT) tolerant ability were investigated for their capabilities in degrading TBT at sublethal concentration. The distribution of TBT and its degraded products dibutyltin (DBT) and monobutyltin (MBT) in the incubation medium, extracellular surface and intracellular fraction were monitored during an exposure period of 14 days. Results showed that biosorption of TBT by the algal cell wall was the major mechanism in reducing 40% of the initial TBT from the medium in the first 2 days. The half-life of TBT incubated with C. vulgaris was 60 h while that of Chlorella sp. was 80 h. The occurrence of DBT at Day 1 in the culture medium provided direct evidence to the biodegradation of TBT by both Chlorella species. At the end of the experimental period, 27 and 41% of the original TBT were recovered as DBT and MBT in cultures of C. vulgaris, respectively. In contrast, DBT appeared to be the only degradation product of Chlorella sp. and only 26% of the original TBT was transformed to DBT. Despite the same genus, TBT was debutylated to a greater extent to MBT by C. vulgaris, while DBT was the end degradation product by Chlorella sp. The capability of such debutylating process therefore accounted for the higher tolerant ability of C. vulgaris than Chlorella sp.     

Copper and zinc induction of lipid peroxidation and effects on antioxidant enzyme activities in the microalga Pavlova viridis (Prymnesiophyceae)

The metal-induced lipid peroxidation and response of antioxidative enzymes have been investigated in the marine microalga Pavlova viridis to understand the mechanisms of metal resistance in algal cells. We have analyzed superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) activities and glutathione (GSH) contents in microalgal cells grown at different concentrations of copper and zinc. In response to each metal, lipid peroxidation was enhanced with the increase of concentrations, as an indication of the oxidative damage caused by metal concentration assayed in the microalgae cells. Exposure of P. viridis to the two metals caused changes in enzyme activities in a different manner, depending on the metal assayed: after copper treatments, total SOD activity was enhanced, while it was reduced after zinc exposure. Copper and zinc stimulated the activities of CAT and GSH whereas GPX showed a remarkable increase in activity in response to copper treatments and decrease after zinc treatments. These results suggest that an activation of some antioxidant enzymes was enhanced to counteract the oxidative stress induced by the two metals.     

Microalgae biosynthesis of silver nanoparticles for application in the control of agricultural pathogens

The occurrence of diseases in cultivars has caused significant losses in global food production. The advancement of nanobiotechnology makes it possible to obtain new products to be used in the control of pathogens in cultivars. Silver nanoparticles can be synthesized by microalgae and are widely known for their antimicrobial activity. In addition, the biomass produced in microalgal culture for the biosynthesis of the nanoparticles also demonstrates antimicrobial properties, as it can increase the antibacterial and antifungal potential of the silver nanoparticles. In this context, this article addresses the use of microalgae to biosynthesize silver nanoparticles simultaneously with biomass production. In addition, we demonstrate the antimicrobial potential of these nanomaterials, as well as of the microalgal biomass produced in biosynthesis, to use in the control of pathogens in agriculture.     

Toxicity of Heavy Metals on Scenedesmus quadricauda (Turp.) de Brébisson in Batch Cultures (7 pp)

Background Aquatic environments are often exposed to various pollutants like heavy metals that are released from industrial, agricultural and domestic wastes. Emissions of heavy metals can then enter all ecosystems and bring about severe problems in plants, especially algae depending on the concentrations of a given element. The objective of the investigation presented is to detect toxic effects due to some heavy metals in the biomass of green alga Scenedesmus quadricauda. Methods All experiments were conducted with axenic cultures of the green alga Scenedesmus quadricauda (Turp.) de Brebisson (Chlorococcales, Chlorophyta). The cultures were grown in Chu-no.10 medium, and optimum physical and chemical growth conditions were provided to get higher growth rates and lower doubling times of cells. Growth of the micro algal cultures was measured on a daily schedule by counting cultures and determination of chlorophyll-a. The sterile-filtered heavy metal concentration solutions (Cd, Pb and Cu) were prepared and added as stock solutions of their salts (CdCl2*2.5 H2O, Pb(NO3)2*H2O and CuSO4*5 H2O). Results and Discussion The growth decreased gradually with the alga exposed to Cd at 0.05, 0.1, and 0.5 mg/L in comparison with the control whereas 1 mg/L Cd2+ had a clearly destructive effect. The growth was decreased with Pb at 15, 20 and 25 mg/L gradually, while at 30 mg/L the effect was more pronounced. When Cu was used, the growth was increased gradually at 0.5, 1 and 1.5 mg/L and vice versa at 2 mg/L. Conclusion There were differences in toxic effects of the metals with different metal as well as their concentrations and the time of exposure.     

Copper complexing properties of dissolved organic materials exuded by the freshwater microalgae Scenedesmus acuminatus (Chlorophyceae)

Dissolved organic materials released by the freshwater microalgae Scenedesmus acuminatus were fractionated into low- and high-molecular weight materials, which were investigated for their capacity to bind copper. The high-molecular weight material was also investigated for its monosaccharide composition and is further discussed in relation to the copper binding property. S. acuminatus was grown in batch cultures under laboratory controlled conditions and harvested at the beginning of stationary growth phase when exuded organic materials were obtained. Copper-complexing property of the total exuded organic materials and exopolysaccharides before and after freeze-drying was evaluated by complexometric titrations and Scatchard Plot Analysis of the titration data. The results revealed the presence of two copper-complexing ligands in the total exuded material, but only one in the exopolysaccharide. Stronger copper-complexing ligands are associated to low molecular weight compounds (LogK'1=7.3, LogCL1=-5.6; LogK'2=6.3, LogCL2=-5.1), whereas weaker ligands to the high molecular weight fraction (LogK'2=6.4, LogCL2=-5.6). Although freeze-drying the polymeric organic material (exopolysaccharide) may result in conformational changes of the molecule, no effect on copper-complexing properties was detected. Gas chromatography was used to evaluate the monosaccharide composition of the microalgal exopolysaccharide, which detected high content of mannose and 12% of acid monosaccharides.     

Biomonitoring in a clean and a multi-contaminated estuary based on biomarkers and chemical analyses in the endobenthic worm Nereis diversicolor

Relationships between biochemical and physiological biomarkers (acetylcholinesterase [AChE], catalase, and glutathione S-transferase [GST] activities, thiobarbituric acid reactive substances, glycogen, lipids and proteins) and accumulated concentrations of contaminants (polychlorinated biphenyls [PCBs], polycyclic aromatic hydrocarbons and metals) were examined in the keystone species Nereis diversicolor. The chemical analyses of worms and sediments allowed the designation of the Seine estuary and the Authie estuary as a polluted and relatively clean site respectively. Worms from the Seine estuary exhibited higher GST and lower AChE activities. Generally, larger worms had higher concentrations of energy reserves. Principal component analyses clearly highlighted intersite differences: in the first plan, GST activities and chemical concentrations were inversely related to concentrations of energy reserves; in the second one, PCB concentrations and AChE activity were inversely related. Depleted levels of energy reserves could be a consequence of combating toxicants and might predict effects at higher levels of biological organization. The use of GST and AChE activities and energy reserve concentrations as biomarkers is validated in the field in this keystone species.     

Zinc acclimation and its effect on the zinc tolerance of Raphidocelis subcapitata and Chlorella vulgaris in laboratory experiments.

The effect of zinc acclimation of Raphidocelis subcapitata (syn. Selenastrum capricornutum) and Chlorella vulgaris on their sensitivity towards this metal was examined in a series of laboratory experiments. These two commonly used algal species were acclimated to 65 microg Zn/l and changes in zinc tolerance were monitored using standard growth inhibition tests. The chemically defined ISO medium was used as a control culture medium. Both species demonstrated a maximum increase in zinc tolerance of a factor of 3 after 100 days of acclimation. Shifts in the shape of the concentration-response curve due to acclimation were observed for R. subcapitata. Compared to non-acclimated algae, acclimated R. subcapitata exhibited higher growth rates in all zinc treatments as well as in the controls. This suggests that the use of ISO-medium results in sub-optimal growth due to zinc deficiency. These effects could not be demonstrated for C. vulgaris. The zinc tolerance of both species decreased significantly one week after returning the acclimated algae to control (ISO) medium. 72hEC50 values based on growth rate were two to four times higher than those calculated using biomass measurements. Algal toxicity test results, particularly if used for metal risk assessments, must not be conducted using nutrient deficient media.     

Toxicity and bioaccumulation of copper in three green microalgal species

The effective concentrations of copper on the inhibition of the growth of Scenedesmus obliquus, Chlorella pyrenoidosa and Closterium lunula at 96 h (96 h EC50) were determined to be 50, 68 and 200 microg/l, respectively. The low initial bioaccumulation of Cu by C. lunula was found to be responsible for its tolerance to Cu. The amount of Cu accumulated by all three microalgae reached the maximum value and decreased quickly after the peak followed by a slow decrease over the next 6 d. Bioaccumulation of Cu by C. lunula was directly proportional to the initial Cu concentration. After reaching the first peak after 1 d, the bioconcentration factor of Cu by microalgae declined to its minimum value during the exponential growth phase but increased in the stationary growth phase again. This indicates that desorption of Cu from microalgae was higher during the exponential growth phase but lower in the stationary growth phase. Smaller microalgae with low 96 h EC50 values are more efficient in removing Cu from wastewater.     

Effect of herbicides on glutathione S-transferases in the earthworm, <Emphasis Type="Italic">Eisenia fetida</Emphasis>

AIM AND BACKGROUND: Earthworms have been studied as a readily available, easily maintainable and cheap test species for assessing chemical pollution, and may be an alternative to in vivo rodent bioassays. The current investigation aims to characterize detoxification enzymes in Eisenia fetida and stress response against two herbicides with different modes of action, namely, fenoxaprop and metolachlor. METHODS: Herbicides were applied to soil containing earthworms. Animals were then collected, sacrificed and shock-frozen. Extracted protein was analyzed for glutathione S-transferase (GST) activity using CDNB (1-chloro-2,4-dinitrobenzene), DCNB (1,2-dichloro-4-nitrobenzene), pNBC (p-nitrobenzylchloride), PNOBC (p-nitrobenz-o-ylchloride) and selected herbicides. GST isoenzymes were partially purified by affinity chromatography and molecular weights were estimated by SDS-PAGE. RESULTS: In E. fetida protein extracts, GST activity towards model compounds ranked as CDNB>DCNB>PNBOC>PNBC. Fluorodifen was not conjugated at all, but fenoxaprop and metolachlor were conjugated at low rates. Furthermore, the GST isoenzyme pattern changed during the incubation with herbicides, either due to stress or as a defense reaction. After incubation with monochlorobimane, a strong fluorescence of the intestinal tract and the intersegments was observed, indicating organ-specific GST induction. DISCUSSION: According to the author's knowledge, here, for the first time, evidence is presented that E. fetida GST are also capable of conjugating a wider range of xenobiotic substrates. Different forms of GST were observed and changes in GST isoforms due to the herbicide treatment were also noticed. GST conjugation rates varied between different herbicides used in this experiment. It might be assumed that herbicides may well be detoxified by earthworms, to a certain extent, but that they are also potent stress factors influencing the detoxification system of the animal. High doses or long exposure might lead to deleterious effects on earthworms and limit their survival rate. The use of the animals as bioindicators for herbicides and herbicide residues seems very promising, but is surely influenced by the lack of detoxification for some compounds. CONCLUSIONS: Conjugation of several xenobiotics with model substances and herbicides is proven in the earthworm E. fetida. However, E. fetida has only limited capabilities of detoxifying herbicidal compounds. Different isoforms of GST were involved and altered in their activity after treatment. RECOMMENDATIONS AND PERSPECTIVES: The accumulation of GS-conjugates and their determination via fluorescence microscopy is a quick and secure, additional marker for exposure that should be further developed to complement existing biotests. The described methods and endpoints might help to understand the complex reaction of earthworms towards herbicides and lead to an adapted test methodology.     

Carbon dioxide capture strategies from flue gas using microalgae: a review

Global warming and pollution are the twin crises experienced globally. Biological offset of these crises are gaining importance because of its zero waste production and the ability of the organisms to thrive under extreme or polluted condition. In this context, this review highlights the recent developments in carbon dioxide (CO₂) capture from flue gas using microalgae and finding the best microalgal remediation strategy through contrast and comparison of different strategies. Different flue gas microalgal remediation strategies discussed are as follows: (i) Flue gas to CO₂ gas segregation using adsorbents for microalgal mitigation, (ii) CO₂ separation from flue gas using absorbents and later regeneration for microalgal mitigation, (iii) Flue gas to liquid conversion for direct microalgal mitigation, and (iv) direct flue gas mitigation using microalgae. This work also studies the economic feasibility of microalgal production. The study discloses that the direct convening of flue gas with high carbon dioxide content, into microalgal system is cost-effective.     

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.     

Evaluation of different algal species sensitivity to mercury and metolachlor by PAM-fluorometry.

In this study, the pulse-amplitude-modulation (PAM)-fluorometric method was used to evaluate the difference in the sensitivity to mercury (Hg) and metolachlor of six algal species: Ankistrodesmus falcatus, Selenastrum capricornutum, Chlorella vulgaris, Nannoplankton (PLS), Microcystis aeruginosa and Pediastrum biwae. We found that the fluorescence parameters (phiM, the maximal photosystem II (PSII) quantum yield, phi'M, the operational PSII quantum yield at steady state of electron transport, Q(P), the photochemical quenching value, and Q(N), the non-photochemical quenching value) were appropriate indicators for inhibitory effects of mercury but only phi'M and Q(N) were useful for metolachlor. The examined algal species showed very different levels of sensitivity to the effect of Hg and of metolachlor. The most sensitive species to Hg and metolachlor were respectively M. aeruginosa and A. falcatus, while the least sensitive were C. vulgaris and P. biwae. We interpreted these differences by the action mode of pollutants and by the different metabolism properties and morphological characteristics between algal species. These results related to fluorescence parameters may offer useful tool to be used in bioassay for different pollutants. Heterogeneous algal sensitivity to the same pollutant suggests the need to use a battery of species to evaluate the effects of mixtures of pollutants in aquatic systems.     

Aquatic toxicity of glyphosate-based formulations: comparison between different organisms and the effects of environmental factors

Glyphosate-based herbicides (e.g. Roundup) are extensively used in the aquatic environment, but there is a paucity of data on the toxicity of the formulated products and the influences by environmental factors. In this study, the acute toxicity of technical-grade glyphosate acid, isopropylamine (IPA) salt of glyphosate, Roundup and its surfactant polyoxyethylene amine (POEA) to Microtox bacterium (Vibrio fischeri), microalgae (Selenastrum capricornutum and Skeletonema costatum), protozoa (Tetrahymena pyriformis and Euplotes vannus) and crustaceans (Ceriodaphnia dubia and Acartia tonsa) was examined and the relative toxicity contributions of POEA to Roundup were calculated. The effects of four environmental factors (temperature, pH, suspended sediment and algal food concentrations) on the acute toxicity of Roundup to C. dubia were also examined. Generally, the toxicity order of the chemicals was: POEA>Roundup>glyphosate acid>IPA salt of glyphosate, while the toxicity of glyphosate acid was mainly due to its high acidity. Microtox bacterium and protozoa had similar sensitivities towards Roundup toxicity (i.e. IC50 from 23.5 to 29.5 mg AE/l). In contrast, microalgae and crustaceans were 4-5 folds more sensitive to Roundup toxicity than bacteria and protozoa. Except photosynthetic microalgae, POEA accounted for more than 86% of Roundup toxicity and the toxicity contribution of POEA was shown to be species-dependent. Increase in pH (6-9) and increase of suspended sediment concentration (0-200 mg/l) significantly increased the toxicity of Roundup to C. dubia, but there were no significant effects due to temperature change and food addition.