Green Alga Pseudokirchneriella Subcapitata Research Articles

Life and death of Pseudokirchneriella subcapitata: physiological changes during chronological aging.

The green alga Pseudokirchneriella subcapitata is widely used in ecotoxicity assays and has great biotechnological potential as feedstock. This work aims to characterize the physiology of this alga associated with the aging resulting from the incubation of cells for 21days, in the OECD medium, with continuous agitation and light exposure, in a batch mode. After inoculation, cells grow exponentially during 3days, and the culture presents a typical green color. In this phase, "young" algal cells present, predominantly, a lunate morphology with the chloroplast occupying a large part of the cell, maximum photosynthetic activity and pigments concentration, and produce starch as a reserve material. Between the 5th and the 12th days of incubation, cells are in the stationary phase. The culture becomes less green, and the cells stop dividing (≥ 99% have one nucleus) and start to age. "Old" algal cells present chloroplast shrinkage, an abrupt decline of chlorophylls content, and photosynthetic capacity (Fv/Fm and ɸPSII), accompanied by a degradation of starch and an increase of neutral lipids content. The onset of the death phase occurs after the 12th day and is characterized by the loss of cell membrane integrity of some algae (cell death). The culture stays, progressively, yellow, and the majority of the population (~93%) is composed of live cells, chronologically "old," with a significant drop in photosynthetic activity (decay > 75% of Fv/Fm and ɸPSII) and starch content. The information here achieved can be helpful when exploring the potential of this alga in toxicity studies or in biotechnological applications. KEY POINTS: • Physiological changes of P. subcapitata with chronological aging are shown • "Young" algae exhibit a semilunar shape, high photosynthetic activity, and accumulated starch • "Old"-live algae show reduced photosynthetic capacity and accumulated lipids.

Ecological Impact of End-of-Life-Tire (ELT)-Derived Rubbers: Acute and Chronic Effects at Organism and Population Levels.

Considering the large amount of tires that reach the end of life every year, the aim of this study was the evaluation of both acute and chronic effects of end-of-life-tire (ELT)-derived rubber granules (ELT-dg) and powder (ELT-dp) on a freshwater trophic chain represented by the green alga Pseudokirchneriella subcapitata, the crustacean Daphnia magna and the teleost Danio rerio (zebrafish). Adverse effects were evaluated at the organism and population levels through the classical ecotoxicological tests. Acute tests on D. magna and D. rerio revealed a 50% effect concentration (EC50) > 100.0 mg/L for both ELT-dg and ELT-dp. Chronic exposures had a lowest observed effect concentration (LOEC) of 100.0 mg/L for both ELT-dg and ELT-dp on P. subcapitata grow rate and yield. LOEC decreased in the other model organisms, with a value of 9.8 mg/L for D. magna, referring to the number of living offspring, exposed to ELT-dg suspension. Similarly, in D. rerio, the main results highlighted a LOEC of 10.0 mg/L regarding the survival and juvenile weight parameters for ELT-dg and a LOEC of 10.0 mg/L concerning the survival and abnormal behavior in specimens exposed to ELT-dp. Tested materials exhibited a threshold of toxicity of 9.8 mg/L, probably a non-environmental concentration, although further investigations are needed to clarify the potential ecological impact of these emerging contaminants.

Exposure of the alga Pseudokirchneriella subcapitata to environmentally relevant concentrations of the herbicide metolachlor: Impact on the redox homeostasis

This study investigated the effect of the herbicide metolachlor (MET) on the redox homeostasis of the freshwater green alga Pseudokirchneriella subcapitata. At low MET concentrations (≤40 μg L−1), no effects on algal cells were detected. The exposure of P. subcapitata to 45–235 μg L−1 MET induced a significant increase of reactive oxygen species (ROS). The intracellular levels of ROS were particularly increased at high (115 and 235 μg L−1) but environmentally relevant MET concentrations. The exposure of algal cells to 115 and 235 μg L−1 MET originated a decrease in the levels of antioxidants molecules (reduced glutathione and carotenoids) as well as a reduction of the activity of scavenging enzymes (superoxide dismutase and catalase). These results suggest that antioxidant (non-enzymatic and enzymatic) defenses were affected by the excess of MET. As consequence of this imbalance (ROS overproduction and decline of the antioxidant system), ROS inflicted oxidative injury with lipid peroxidation and damage of cell membrane integrity. The results provide further insights about the toxic modes of action of MET on a non-target organism and emphasize the relevance of toxicological studies in the assessment of the impact of herbicides in freshwater environments.

Self-assembly of chlorin-e6 on γ-Fe2O3 nanoparticles: Application for larvicidal activity against Aedes aegypti

Aedes aegypti mosquitos are widespread vectors of several diseases and their control is of primary importance for biological and environmental reasons, and novel safe insecticides are highly desirable. An eco-friendly photosensitizing magnetic nanocarrier with larvicidal effects on Aedes aegypti was proposed. The innovative core-shell hybrid nanomaterial was synthesized by combining peculiar magnetic nanoparticles (called Surface Active Maghemite Nanoparticles - SAMNs, the core) and chlorin-e6 as photosensitizer (constituting the shell) via self-assembly in water. The hybrid nanomaterial (SAMN@chlorin) was extensively characterized and tested for the photocidal activity on larvae of Aedes aegypti. The SAMN@chlorin core-shell nanohybrid did not present any toxic effect in the dark, but, upon light exposure, showed a higher photocidal activity than free chlorin–e6. Moreover, the eco-toxicity of SAMN@chlorin was determined in adults and neonates of Daphnia magna, where delayed toxicity was observed only after prolonged (≥4 h) exposure to intense light, on the green alga Pseudokirchneriella subcapitata and on the duckweed Lemna minor on which no adverse effects were observed. The high colloidal stability, the physico-chemical robustness and the magnetic drivability of the core-shell SAMN@chlorin nanohybrid, accompanied by the high photocidal activity on Aedes aegypti larvae and reduced environmental concerns, can be proposed as a safe alternative to conventional insecticides.

2-Hydroxynaphthaldehyde chitosan schiff-base; new complexes, biosorbent to remove cadmium(II) ions from aqueous media and aquatic ecotoxicity against green alga Pseudokirchneriella subcapitata

Chemically modified chitosan (CS) has been produced with the aim of forming a Schiff-base with 2-hydroxynaphthaldehyde (2CS-Hhnal). Complexes of 2CS-Hhnal with Ni(II), Cu(II), Zn(II), Pd(II), Ag(I), Cd(II) and Hg(II) have been synthesized and characterized by various spectroscopic methods (FT-IR, Raman, UV–vis reflectance, solid-state 13C NMR and XRD) and morphological (SEM, EDX), elemental analysis, TGA and magnetic measurements. The morphology and structure of CS are different from those of the 2CS-Hhnal Schiff-base. In the complexes, the uninegative bidentate ligand (2CS-hnal−) is coordinated to the metal ions through the azomethine-N and deprotonated hydroxy-O atoms without any participation from the glucose amine hydroxy-O atoms. Adsorption is considered to be a potential cost-effective method for the removal of heavy metal ions from aqueous media and by incorporating 2CS-Hhnal, we found that the adsorption of Cd(II) ions was greatly enhanced when compared to adsorption with free CS itself. The optimum experimental parameters for this process are discussed in this paper, including the initial pH, amount of sorbent, contact time and Cd(II) ion concentration. The experimental data were fitted well by a pseudo-second-order kinetics model, while the equilibrium data could be accounted for by a Langmuir model. The desorption process with EDTA and HNO3 (0.1 mol L−1) illustrates the regeneration ability of 2CS-Hhnal without any significant loss of its initial properties throughout three adsorption–desorption cycles. Compared to commercially expensive sorbents, the non-toxic, biodegradable 2CS-Hhnal material could prove to be an attractive, alternative material for the removal of Cd(II) ions from wastewater. The amount of Cd(II) ions adsorbed onto 2CS-Hhnal was determined by atomic absorption spectroscopy and from the intensity of the Raman-active azomethine (CN) vibration. In addition, the aquatic ecotoxicity of free 2CS-Hhnal and its complexes against green alga Pseudokirchneriella subcapitata was evaluated.

Evaluation of acute and chronic ecotoxicity of cyclophosphamide, ifosfamide, their metabolites/transformation products and UV treated samples

Cyclophosphamide (CP) and Ifosfamide (IF) are two nitrogen mustard drugs widely prescribed in cancer therapy. They are continuously released via excreta into hospital and urban wastewaters reaching wastewater treatment plants. Although CP and IF, their metabolites and transformation products (TPs) residues have been found in the aquatic environment from few ng L-1 to tens of μg L-1, their environmental toxic effects are still not well known. The present study aimed to investigate the acute and chronic ecotoxicity of CP and IF and their commercially available human metabolites/TPs, i.e. carboxy-CP, Keto-CP and N-dechloroethyl-CP on different organisms of the aquatic trophic chain. The experiments were performed using the green alga Pseudokirchneriella subcapitata, the rotifer Brachionus calyciflorus and the crustaceans Thamnocephalus platyurus and Ceriodaphnia dubia. Moreover, to assess the treatment conditions in regards to parent compound removal and formation of new TPs, CP and IF were UV- irradiated for 6h, 12h, 24h, 36h and 48h, followed by toxicity evaluation of treated samples by algae, rotifers and crustaceans. Between the parent compounds, IF resulted as more toxic drug under tested conditions, exerting both acute and chronic effects especially on C.dubia (LC50:196.4mgL-1, EC50:15.84mgL-1). Among the tested metabolites/TPs, only carboxy-CP inhibited the reproduction in the rotifer. However, LOEC and NOEC values were calculated for CP and IF for all organisms. In addition, despite a low degradation of CP (28%) and IF (36%) after 48h UV-irradiation, statistically significant effect differences (p<0.05) from not-irradiated and irradiated samples were observed in both acute and chronic assays, starting from 6h UV-irradiation. Our results suggest that the toxic effects found in the aquatic organisms may be attributable to interactions between the parent compounds and their metabolites/TPs.

Cell reproductive patterns in the green alga Pseudokirchneriella subcapitata (=Selenastrum capricornutum) and their variations under exposure to the typical toxicants potassium dichromate and 3,5-DCP.

Pseudokirchneriella subcapitata is a sickle-shaped freshwater green microalga that is normally found in unicellular form. Currently, it is the best known and most frequently used species of ecotoxicological bioindicator because of its high growth rate and sensitivity to toxicants. However, despite this organism’s, our knowledge of its cell biology—for example, the patterns of nuclear and cytoplasmic division in the mitotic stage—is limited. Although it has been reported that P. subcapitata proliferates by popularity forming four daughter cells (autospores) through multiple fission after two nuclear divisions, here, we report two additional reproductive patterns by which two autospores are formed by binary fission (“two-autospore type”) and eight autospores are formed by multiple fission (“eight-autospore type”). Moreover, we found that cell reproductive patterns differed markedly with the culture conditions or with exposure to either of two typical toxicants, potassium dichromate (K2Cr2O7) and 3,5-dichlorophenol (3,5-DCP). The eight-autospore type occurred at the highest frequency in the early phase of culture, but it disappeared under 3,5-DCP at 2.0 mg/L. Under 0.3 mg/L K2CrO7 (Cr(VI)) the eight-autospore type took substantially longer to appear than in control culture. The two-autospore type occurred only in the late phase of culture. To our knowledge, this is the first detailed evaluation of the reproductive patterns of P. subcapitata, which changed dramatically in the presence of toxicants. These findings suggest that observation of the reproductive patterns of P. subcapitata will help to elucidate different cell reactions to toxicants.

Toxicity of waters from the Rochester Embayment Area of Concern to the plankton species Pseudokirchneriella subcapitata and Ceriodaphnia dubia

The lower Genesee River and Rochester Embayment of Lake Ontario are a designated Area of Concern (AOC) under the binational Great Lakes Water Quality Agreement. The “degradation of phytoplankton and zooplankton populations” or plankton Beneficial Use Impairment (BUI) was classified as unknown and in need of further assessment in this AOC because water quality data suggested plankton communities could be effected and community data were either unavailable or indicated impacts. The plankton BUI may now be obsolete because local contaminant sources have been largely eliminated. The present study was conducted between July 2013 and August 2014 to assess the BUI-removal criteria: “AOC plankton bioassays confirm that toxicity in ambient waters (i.e., no growth inhibition) is not significantly higher than comparable non-AOC controls”. Acute and chronic toxicity of waters from 13 sites were quantified seasonally using standardized bioassays with the green alga Pseudokirchneriella subcapitata and water flea Ceriodaphnia dubia to test the hypothesis that toxicity of waters from AOC sites was not higher than that of waters from comparable non-AOC reference sites. Survival and reproduction of C. dubia did not differ significantly between site types, systems, or months. The growth of P. subcapitata did not differ between site types, but differed among systems and months. All results indicate that waters from AOC sites were no more toxic to both plankton species than waters from reference sites. Assuming test species represent natural plankton assemblages, water quality should not negatively affect survival and growth of resident plankton populations in the Rochester Embayment AOC.

Toxicity of the mixture of selected antineoplastic drugs against aquatic primary producers.

The residues of antineoplastic drugs are considered as new and emerging pollutants in aquatic environments. Recent experiments showed relatively high toxicity of 5-fluorouracil (5-FU), imatinib mesylate (IM), etoposide (ET) and cisplatin (CP) that are currently among most widely used antineoplastic drugs, against phytoplankton species. In this study, we investigated the toxic potential of the mixture of 5-FU + IM + ET against green alga Pseudokirchneriella subcapitata and cyanobacterium Synechococcus leopoliensis, and the stability and sorption of these drugs to algal cells. Toxic potential of the mixture was predicted by the concepts of 'concentration addition' and 'independent action' and compared to the experimentally determined toxicity. In both test species, the measured toxicity of the mixture was at effects concentrations EC10-EC50 higher than the predicted, whereas at higher effect concentration (EC90), it was lower. In general, P. subcapitata was more sensitive than S. leopoliensis. The stability studies of the tested drugs during the experiment showed that 5-FU, IM and CP are relatively stable, whereas in the cultures exposed to ET, two transformation products with the same mass as ET but different retention time were detected. The measurements of the cell-linked concentrations of the tested compounds after 72h exposure indicated that except for CP (1.9% of the initial concentration), these drugs are not adsorbed or absorbed by algal cells. The results of this study showed that in alga and cyanobacteria exposure to the mixture of 5-FU + ET + IM, in particular at low effect concentration range, caused additive or synergistic effect on growth inhibition, and they suggest that single compound toxicity data are not sufficient for the proper toxicity prediction for aquatic primary producers.

Phytotoxicity and genotoxicity assessment of imazethapyr herbicide using a battery of bioassays.

The imazethapyr herbicide (formulation Verosil(®)) was evaluated for phytotoxicity and genotoxicity using a battery of bioassays: (1) the growth inhibition of the green alga Pseudokirchneriella subcapitata, (2) the root growth and germination of the higher plant Lactuca sativa, (3) the genetic damage using the Salmonella/microsome test, and (4) the aneugenic and clastogenic effects on Allium cepa. The Verosil(®) formulation was highly toxic to the non-target green alga (median effective concentration (EC50) = 1.05 ± 0.05 mg active ingredient (a.i.) L(-1)), and concentrations above 10 mg a.i. L(-1) inhibited root elongation in lettuce: relative growth index (RGI) between 0.28 ± 0.01 and 0.66 ± 0.10. No genotoxic effect was observed in S almonella typhimurium at 100 mg a.i. L(-1), either with or without the microsomal fraction. However, significant differences in the frequency of chromosomal aberrations in anaphases and telophases (bridges, chromosome fragments, and vagrants) were observed in A. cepa at concentrations between 0.01 and 1 mg a.i. L(-1) with respect to the control. The frequencies of micronuclei showed significant differences with respect to the control at concentrations between 0.001 and 0.1 mg a.i. L(-1). A very high mitotic index (MI = 93.8 ± 5.8) was observed associated with a high number of cells in the prophase stage at 100 mg a.i. L(-1), indicating cytotoxicity. These results showed that imazethapyr is toxic to the non-target populations in both aquatic and terrestrial ecosystems. This herbicide might also exert clastogenic and aneugenic mitotic damage in higher plants. Therefore, the imazethapyr formulation may constitute an environmental risk to plants.

Responses of the alga Pseudokirchneriella subcapitata to long-term exposure to metal stress

The green alga Pseudokirchneriella subcapitata has been widely used in ecological risk assessment, usually based on the impact of the toxicants in the alga growth. However, the physiological causes that lead algal growth inhibition are not completely understood. This work aimed to evaluate the biochemical and structural modifications in P. subcapitata after exposure, for 72h, to three nominal concentrations of Cd(II), Cr(VI), Cu(II) and Zn(II), corresponding approximately to 72h-EC10 and 72h-EC50 values and a high concentration (above 72h-EC90 values). The incubation of algal cells with the highest concentration of Cd(II), Cr(VI) or Cu(II) resulted in a loss of membrane integrity of ~16, 38 and 55%, respectively. For all metals tested, an inhibition of esterase activity, in a dose-dependent manner, was observed. Reduction of chlorophyll a content, decrease of maximum quantum yield of photosystem II and modification of mitochondrial membrane potential was also verified. In conclusion, the exposure of P. subcapitata to metals resulted in a perturbation of the cell physiological status. Principal component analysis revealed that the impairment of esterase activity combined with the reduction of chlorophyll a content were related with the inhibition of growth caused by a prolonged exposure to the heavy metals.

Elimination and detoxification of fungicide miconazole and antidepressant sertraline by manganese peroxidase-dependent lipid peroxidation system

The imidazole fungicide miconazole (MCZ) and the antidepressant sertraline (SER) have the potential to adversely affect aquatic organisms when they are released into the environment. To assess if they could be broken down, MCZ and SER were treated with laccase and manganese peroxidase (MnP) from lignin-degrading white-rot fungi. The MnP-dependent lipid peroxidation system with Tween 80 containing unsaturated fatty acid was effective in eliminating MCZ and SER; the MnP-Tween 80 system helped eliminate 88% of MCZ and 85% of SER after 24 h of treatment. Furthermore, this system eliminated a metabolite of SER (desmethylsertraline or DSER), and no differences between the elimination rates of DSER and SER were observed throughout the treatment period. It was also confirmed that the MnP-Tween 80 system caused a complete loss of growth inhibition by MCZ of the gram-positive bacterium Bacillus subtilis and reduced the growth inhibition by SER of the green alga Pseudokirchneriella subcapitata by 78% after 24 h of treatment. On the other hand, no appreciable elimination of MCZ and SER was obtained by treatment with laccase, MnP, or the laccase-mediator system with 1-hydroxybenzotriazole.

Use of a fluorescence-based approach to assess short-term responses of the alga Pseudokirchneriella subcapitata to metal stress

This work explores the use of fluorescent probes to evaluate the responses of the green alga Pseudokirchneriella subcapitata to the action of three nominal concentrations of Cd(II), Cr(VI), Cu(II) and Zn(II) for a short time (6 h). The toxic effect of the metals on algal cells was monitored using the fluorochromes SYTOX Green (SG, membrane integrity), fluorescein diacetate (FDA, esterase activity) and rhodamine 123 (Rh123, mitochondrial membrane potential). The impact of metals on chlorophyll a (Chl a) autofluorescence was also evaluated. Esterase activity was the most sensitive parameter. At the concentrations studied, all metals induced the loss of esterase activity. SG could be used to effectively detect the loss of membrane integrity in algal cells exposed to 0.32 or 1.3 μmol L−1 Cu(II). Rh123 revealed a decrease in the mitochondrial membrane potential of algal cells exposed to 0.32 and 1.3 μmol L−1 Cu(II), indicating that mitochondrial activity was compromised. Chl a autofluorescence was also affected by the presence of Cr(VI) and Cu(II), suggesting perturbation of photosynthesis. In conclusion, the fluorescence-based approach was useful for detecting the disturbance of specific cellular characteristics. Fluorescent probes are a useful diagnostic tool for the assessment of the impact of toxicants on specific targets of P. subcapitata algal cells.

Natural dissolved organic matter mobilizes Cd but does not affect the Cd uptake by the green algae Pseudokirchneriella subcapitata (Korschikov) in resin buffered solutions

Natural dissolved organic matter (DOM) can have contrasting effects on metal bioaccumulation in algae because of complexation reactions that reduce free metal ion concentrations and because of DOM adsorption to algal surfaces which promote metal adsorption. This study was set up to reveal the role of different natural DOM samples on cadmium (Cd) uptake by the green algae Pseudokirchneriella subcapitata (Korschikov). Six different DOM samples were collected from natural freshwater systems and isolated by reverse osmosis. In addition, one 13C enriched DOM sample was isolated from soil to trace DOM adsorption to algae. Algae were exposed to standardized solutions with or without these DOM samples, each exposed at equal DOM concentrations and at equal non-toxic Cd2+ activity (∼4nM) that was buffered with a resin. The DOM increased total dissolved Cd by factors 3–16 due to complexation reactions at equal Cd2+ activity. In contrast, the Cd uptake was unaffected by DOM or increased maximally 1.6 fold (13C enriched DOM). The 13C analysis revealed that maximally 6% of algal C was derived from DOM and that this can explain the small increase in biomass Cd. It is concluded that free Cd2+ and not DOM-complexed Cd is the main bioavailable form of Cd when solution Cd2+ is well buffered.

Heterogeneous photocatalysis of moxifloxacin in water: Chemical transformation and ecotoxicity

This work provides new insights on the impact of TiO2/UV catalyzed chemical transformation of moxifloxacin on ecotoxicity effects towards the green alga Pseudokirchneriella subcapitata. The moxifloxacin median effect concentration (EC-50=0.78 [0.56, 1.09] mgL−1), determined in accordance to the OECD 72-h growth inhibition test guideline, was 7 times lower than that of the older and widely used fluoroquinolone ciprofloxacin (EC-50=5.57 [4.86, 6.38] mgL−1).Applying heterogeneous photocatalysis as an advanced oxidation technique to degrade moxifloxacin in aqueous solution decreased the average growth inhibition from 72% to 14% after 150min of treatment. No significant carbon mineralization was observed and liquid chromatography mass spectrometry analysis revealed the formation of 13 degradation products for which a chemical structure could be proposed based on accurate mass determination. Combined chemical and ecotoxicological analysis showed that as long as moxifloxacin is present in the reaction solution, it is the main compound affecting algal growth inhibition. However, also the contribution of the degradation products to the observed ecotoxicity cannot be neglected.Photocatalytically induced modifications of moxifloxacin mainly occur at the diazobicyclo-substituent as ring opening, oxidation into carbonyl groups, and hydroxylation. This results into the formation of more hydrophilic compounds with a decreased biological activity compared with moxifloxacin. The change in lipophilicity, and possibly a modified acid–base speciation, most probably also affect the cell membrane permeation of the degradation products, which might be another factor explaining the observed lower residual ecotoxicity of the photocatalytically treated reaction solutions.

The effect of different polychlorinated biphenyls on two aquatic models, the green alga Pseudokirchneriella subcapitata and the haemocytes from the European abalone Haliotis tuberculata

The present study was conducted to determine the toxicity of different polychlorinated biphenyls (PCBs) on the green algae, Pseudokirchneriella subcapitata and the haemocytes from the European abalone, Haliotis tuberculata. Using the algal growth inhibition test, the green algae median Effective Concentration (EC50) values ranged from 0.34μM for PCB28 to more than 100μM for PCBs 101 and 153. Considering the MTT viability test, the abalone EC50 values ranged from 1.67μM for PCB153 to 89μM for PCB28. Our results in contrast to previous observation in vertebrates did not show significant differences between the dioxin like- and non dioxin like-PCBs toxicities regardless of the model used. However, our results demonstrated that the toxicities of PCBs were species dependent. For example, PCB28 was the most toxic compound for P. subcapitata whereas PCBs 1, 180 and 153 were less toxic for that species. On the contrary, PCB153 was reported as the most toxic for H. tuberculata haemocytes and PCB28 the least toxic. To investigate the mode of action of these compounds, we used an in silico method. Our results suggested that PCBs have a non-specific mode of action (e.g., narcosis) on green algae, and another mode of action, probably more specific than narcosis, was reported for PCBs on the abalone haemocytes.

Algal growth stimulation and toxicity in response to exposure to the new insensitive military high‐nitrogen energetic triaminoguanidinium‐1‐methyl‐5‐nitriminotetrazolate

Triaminoguanidinium-1-methyl-5-nitriminotetrazolate (TAG-MNT) is a nitrogen-rich energetic compound being developed as a potential component of insensitive munition formulations. The purpose of the present study was to assess the toxicity of TAG-MNT to the green alga Pseudokirchneriella subcapitata as well as to determine whether the high N content of TAG-MNT could result in increased algal growth in aquatic systems and potentially contribute to eutrophication using a 96-h algal growth bioassay in N-limited test media. Results were compared with algal exposures to current-use energetics 2,4,6-trinitrotoluene (TNT) and royal demolition explosive (RDX). The TNT exposure resulted in a lowest-observed-adverse-effect concentration (LOAEC) for algal growth of 1.72 mg/L and a 50% inhibition concentration (IC50) and 95% confidence limits of 0.972 mg/L (0.955, 0.973). The RDX algal growth LOAEC was 0.10 mg/L, and the RDX IC50 was 0.635 (0.416, 0.875). Neither TNT nor RDX exposure resulted in stimulation of algal growth. In repeated testing, TAG-MNT exposure resulted in LOAECs of 0.55 and 5.20 mg/L. Stimulation of algal growth was observed at 0.06 mg/L at a mean increase of 163.2% (± 71.7) relative to the control in TAG-MNT test A and at the 0.005 mg/L treatment at a mean increase of 174.3% (± 59.9) in TAG-MNT test B. The authors' results indicate the potential for high-N energetics to significantly stimulate algal growth at low concentrations in N-limited systems.

Algal Population Growth Model Integrated with Toxicokinetics for Ecological Risk Assessment under Time-Varying Pesticide Exposure

ABSTRACT An algal population growth model integrated with toxicokinetics was developed for assessing the effect of pesticides on population dynamics. This model is a simple one-compartment, first-order kinetic model in which toxicity (growth inhibition and mortality) depends on the intracellular effective concentration of a pesticide at a target site. The model's parameters were derived using an experimental study that investigated the effects of pretilachlor, bensulfuron-methyl, pentoxazone, and quinoclamine on the growth, mortality, and subsequent population recovery of the green alga Pseudokirchneriella subcapitata. Modeled and measured trajectories of algal population dynamics agreed well. The effect on population recovery was underestimated by the model that ignored the toxicokinetics. The four tested herbicides had a variety of toxicity characteristics and physicochemical properties, indicating the wide range of the model's applicability. Moreover, the developed model and the obtained model's parameters were extrapolated to predict long-term algal population dynamics under time-varying herbicide exposure. The calculated integral biomass lost compared with the control was considered a quantitative index of the population-level ecological risk. The model's prediction showed that the same exposure level (peak concentration is equivalent to EC50) indicated much different population-level effect depending on the herbicide.

Toxicities of four anti-neoplastic drugs and their binary mixtures tested on the green alga Pseudokirchneriella subcapitata and the cyanobacterium Synechococcus leopoliensis

The residues of anti-neoplastic drugs are new and emerging pollutants in aquatic environments. This is not only because of their increasing use, but also because due to their mechanisms of action, they belong to a group of particularly dangerous compounds. However, information on their ecotoxicological properties is very limited. We tested the toxicities of four anti-neoplastic drugs with different mechanisms of action (5-fluorouracil [5-FU], cisplatin [CDDP], etoposide [ET], and imatinib mesylate [IM]), and some of their binary mixtures, against two phytoplankton species: the alga Pseudokirchneriella subcapitata, and the cyanobacterium Synechococcus leopoliensis. These four drugs showed different toxic potential, and the two species examined also showed differences in their susceptibilities towards the tested drugs and their mixtures. With P. subcapitata, the most toxic of these drugs was 5-FU (EC50, 0.13 mg/L), followed by CDDP (EC50, 1.52 mg/L), IM (EC50, 2.29 mg/L), and the least toxic, ET (EC50, 30.43 mg/L). With S. leopoliensis, the most toxic was CDDP (EC50, 0.67 mg/L), followed by 5-FU (EC50, 1.20 mg/L) and IM (EC50, 5.36 mg/L), while ET was not toxic up to 351 mg/L. The toxicities of the binary mixtures tested (5-FU + CDDP, 5-FU + IM, CDDP + ET) were predicted by the concepts of ‘concentration addition’ and ‘independent action’, and are compared to the experimentally determined toxicities. The measured toxicity of 5-FU + CDDP with P. subcapitata and S. leopoliensis was higher than that predicted, while the measured toxicity of CDDP + ET with both species was lower than that predicted. The measured toxicity of 5-FU + IM with P. subcapitata was higher, and with S. leopoliensis was lower, than that predicted. These data show that these mixtures can have compound-specific and species-specific synergistic or antagonistic effects, and they suggest that single compound toxicity data are not sufficient for the prediction of the aquatic toxicities of such anticancer drug mixtures.

Effect of food density of Pseudokirchneriella subcapitata and Chlorella vulgaris on the population growth rates of four brachionid rotifers

We conducted population growth experiments for 2–4 weeks using 4 species of brachionid rotifers (Brachionus angularis, B. havanaensis, B. rubens, and Plationus patulus), all isolated from Lake Xochimilco, Mexico City, Mexico. The test rotifers were fed 2 species of green algae (Chlorella vulgaris and Pseudokirchneriella subcapitata) at 3 food concentrations (2.9, 5.8, and 11.6 μg dry weight mL−1) for 24 h. Regardless of algal species, the population density of all 4 rotifer species increased with increasing food density; however, the magnitude of growth rate response of the rotifers differed. At comparable food levels, B. havanaensis had far greater abundances in 3 weeks than the other 3 rotifer species. At a concentration of 2.9 μg mL−1 of C. vulgaris, density of B. angularis was about 50 ind. mL−1, but when the food was enhanced to 11.6 μg mL−1, its population was 3 times higher; similar values were obtained when B. angularis was fed P. subcapitata. In comparison, B. havanaensis fed P. subcapitata at the highest food level had population abundances of about 900 ind. mL−1; at the same food level, when fed C. vulgaris, the density was much lower (250 ind. mL−1). Population abundances of B. rubens fed P. subcapitata were, in general, lower than those fed C. vulgaris. This was also the trend for rotifer P. patulus, which had lower abundances when fed P. subcapitata than when fed C. vulgaris. Regardless of food type and concentration, the rate of population increase per day (r) of the tested rotifer species varied from 0.16 to 0.61 d−1.