Enantioselectivity in toxicity and degradation of dichlorprop-methyl in algal cultures

Enantioselectivity in the toxicity and degradation of the herbicide dichlorprop-methyl (2,4-DCPPM) in algal cultures was studied. Enantioselectivity was clearly observed in the toxicity of racemic 2,4-DCPPM and its two enantiomers. R-2,4-DCPPM showed low toxicity to Chlorella pyrenoidosa and Chlorella vulgaris, but higher toxicity to Scenedesmus obliquus. The observed toxicity was ranked: R-2,4-DCPPM>S-2,4-DCPPM>Rac-2,4-DCPPM; the toxicity of R-2,4-DCPPM was about 8-fold higher than that of Rac-2,4-DCPPM. Additionally, 2,4-DCPPM was quickly degraded, in the initial 12 h, and different algae cultures had different enantioselectivity for the 2,4-DCPPM enantiomers. There was no significant enantioselectivity for 2,4-DCPPM in Chlorella vulgaris in the initial 7 h. However, racemic 2,4-DCPPM was degraded by Scenedesmus obliquus quickly, in the initial 4 h, much quicker, in fact, than the S- or R-enantiomers (racemate>R->S-), indicating that the herbicide 2,4-DCPPM was absorbed enantioselectively by Scenedesmus obliquus. The rapid formation of 2,4-DCPP suggested that 2,4-DCPPM adsorbed by algal cells was catalytically hydrolyzed to the free acid, a toxic metabolite. The production rates of 2,4-DCPP were as follows: Scenedesmus obliquus>Chlorella pyrenoidosa>Chlorella vulgaris, consistent with the degradability of 2,4-DCPPM. Scenedesmus obliquus had quick, but different, degradative and uptake abilities for R-, S-, and Rac-2,4-DCPPM. The R- and S- enantiomers were not hydrolyzed in the first 12 h, while both enantiomers were hydrolyzed slowly after that. These results indicate that some physical and chemical properties of compounds are of importance in determining their enantioselective toxicity and degradation. The ester and its metabolite likely played an important role in enantioselective toxicity to the three algae.     

Biosorption of zinc and copper from aqueous solutions by two freshwater green microalgae Chlorella pyrenoidosa and Scenedesmus obliquus

Purpose

The objective of this study was to determine the removal of zinc and copper by two freshwater green microalgae Chlorella pyrenoidosa and Scenedesmus obliquus and to investigate changes of algal ultrastructure and photosynthetic pigment.

Methods

Algal cells were exposed for 8 days to different initial zinc or copper concentrations. Heavy metal concentrations were detected by an atomic absorption spectrophotometer. Algal growth, ultrastructure, and photosynthetic pigment were analyzed by a microplate reader, transmission electron microscope, and spectrophotometer, respectively.

Results

Low zinc and copper concentrations induced increase in algal growth, whereas application of high zinc and copper concentrations suppressed the growth of both algae. High metal concentrations also decreased the photosynthetic pigments and destroyed algal cell ultrastructure. The zinc removal efficiency by both algae increased rapidly during the first day and thereafter remained nearly constant throughout the experiment. The copper removal efficiency by both algae increased slowly during the whole experimental periods. In all cultures, the quantity of both metals removed intracellularly was much lower than the adsorbed quantity on the cell surface.

Conclusions

Both strains of the microalgae had proven effective in removing zinc and copper from aqueous solutions, with the highest removal efficiency being near 100%. In addition, C. pyrenoidosa appeared to be more efficient than S. obliquus for removing copper ions. On the contrary, S. obliquus appeared to be more efficient than C. pyrenoidosa for removing zinc ions.     

Influence of toxicity and dissipation of racemic fenoxaprop and its R-enantiomer in Scenedesmus obliquus suspension by cyclodextrins

Cyclodextrins (CDs), a class of cyclic oligosaccharide molecules containing a variety of chiral centre, are capable of recognizing enantiomeric molecules through the formation of inclusion complexes. In this work, we selected three types of CDs, β-CD and its two derivatives, randomly methylated β-CD (RAMEB) and hydroxypropyl β-CD (HP-β-CD), to evaluate effects on toxicity of racemic fenoxaprop (rac-FA) and its R-enantiomer (R-FA) to freshwater alga Scenedesmus obliquus (S. obliquus) and their dissipation in S. obliquus suspension with and without CDs addition, respectively, in an attempt to get more detail about enantioselective behavior of fenoxaprop acid (FA) in the environment, using CDs as a remediation agent for FA and formulation additive for fenoxaprop-p-ethyl (FE). The significant difference between rac-FA and R-FA was not observed in their acute toxicity to S. obliquus and dissipation in S. obliquus suspension. RAMEB had no effect on either toxicity of FA to S. obliquus or dissipation of FA in S. obliquus suspension, and it also didn't change the extent of enantioselectivity in toxicity of FA to S. obliquus. But the addition of a certain amount of β -CD and HP-β -CD reduced the toxicity of FA to S. obliquus and increased dissipation of FA in S. obliquus suspension, as well as changed the enantioselectivity in toxicity of FA to S. obliquus. The results indicated β-CD and HP-β-CD could be used as a promising agent for remediation of aquatic contamination produced by FA, and RAMEB might be used as potential formulation additives for FE, the parent compound of FA, as RAMEB didn't decrease activity of R-FA and might be environmentally safer than the conventional additives.     

Algicidal activity of Salvia miltiorrhiza Bung on Microcystis aeruginosa—Towards identification of algicidal substance and determination of inhibition mechanism

The present study was to isolate and identify a potent algicidal compound from extract of Salvia miltiorrhiza and study the potential inhibition mechanism on Microcystis aeruginosa. Column chromatography and bioassay-guided fractionation methods were carried out to yield neo-przewaquinone A, which was identified by spectral analysis. The EC50 of neo-przewaquinone A on M. aeruginosa were 4.68 mg L⁻¹. In addition, neo-przewaquinone A showed relatively higher security on Chlorella pyrenoidosa and Scenedesmus obliquus, with the EC50 values of 14.78 and 10.37 mg L⁻¹, respectively. For the potential inhibition mechanisms, neo-przewaquinone A caused M. aeruginosa cells morphologic damage or lysis, increased malondialdehyde content and decreased the soluble protein content, total antioxidant and superoxide dismutase activity, and significantly inhibited three photosynthesis-related genes (psaB, psbD, and rbcL). The results demonstrated the algicidal effect of neo-przewaquinone A on M. aeruginosa and provided the possible inhibition mechanisms.     

Combined effect of polystyrene plastics and triphenyltin chloride on the green algae Chlorella pyrenoidosa

The combined effect of polystyrene (PS) particles and triphenyltin chloride (TPTCl) to the green algae Chlorella pyrenoidosa was studied. The 96 h IC₅₀ of TPTCl to the green algae C. pyrenoidosa was 30.64 μg/L. The toxicity of PS particles to C. pyrenoidosa was size-dependent, with the 96 h IC₅₀ at 9.10 mg/L for 0.55 μm PS but no toxicity observed for 5.0 μm PS. The exposure to 0.55 μm PS led to damage on structure of algal cells, which could in turn cause inhibition on photosynthesis and population growth of the green algae. TPTCl concentrations in test medium were lowered by 15–19% at presence of 0.55 μm PS particles, indicating a reduced bioavailability of TPTCl. In spite of this reduced bioavailability, the presence of PS increased the toxicity of TPTCl, which might be attributed to facilitated uptake of TPTCl by the green algae after the damage of cell structure. The overall results of the present study provided important information on the effect of PS on the bioavailability and toxicity of TPTCl to phytoplankton species.

     

Enantioselective toxic effects and biodegradation of benalaxyl in Scenedesmus obliquus

Enantioselectivity in ecotoxicity and biodegradation of chiral pesticide benalaxyl to freshwater algae Scenedesmus obliquus was studied. The 96 h-EC₅₀ values of rac-, R-(−)-, S-(+)-benalaxyl were 2.893, 3.867, and 8.441 mg L⁻¹, respectively. Therefore, the acute toxicities of benalaxyl enantiomers were enantioselective. In addition, the pigments chlorophyll a and chlorophyll b, antioxidant enzyme activities catalase (CAT) and superoxide dismutase (SOD) as well as lipid peroxide malondialdehyde (MDA) were determined to evaluate the different toxic effects. Chlorophyll a was induced by S-(+)-benalaxyl but inhibited by R-(−)-benalaxyl at 1 mg L⁻¹. Chlorophyll b were both induced at 1 mg L⁻¹, but S-(+)-form was fourfold higher than R-(−)-form. S-(+)-benalaxyl inhibited more CAT activities at 3 mg L⁻¹ and 5 mg L⁻¹, induced less SOD activity and MDA content at 5 mg L⁻¹ than R-(−)-benalaxyl. Based on these data, enantioselectivity occurred in anti-oxidative stress when S. obliquus response to benalaxyl. In the biodegradation experiment, the half-lives of S-(+)-benalaxyl and R-(−)-benalaxyl were 4.07 d and 5.04 d, respectively, resulting in relative enrichment of the R-(−)-form. These results showed that toxic effects and biodegradation of benalaxyl in S. obliquus were enantioselective, and such enantiomeric differences must be taken into consideration in pesticide risk.     

Phytoplankton toxicity of the antibiotic chlortetracycline and its UV light degradation products

Two common freshwater phytoplankton species Microcystis aeruginosa and Scenedesmus obliquus were employed as test organisms to investigate the toxic effects of chlortetracycline widely used in human medicine and veterinary as antibiotic. Toxicity assays were performed into two parts: antibiotic toxicity test and antibiotic degraded products toxicity test. In general, chlortetracycline had significantly toxic effect on population growth and chlorophyll-a accumulation of two phytoplankton. Although M. aeruginosa had ability to grow after exposed to chlortetracycline at 0.5 mg L⁻¹, its photosynthesis function was also disrupted. Compared with the data in two phytoplankton species, the chlorophyceae was more sensitive than the cyanophyceae. The adverse effect on S. obliquus was stronger than that on M. aeruginosa with increasing concentrations. In addition, for M. aeruginosa, regardless of the UV light degradation time, the treated chlortetracycline also had adverse effect on population growth and chlorophyll-a accumulated. The degraded chlortetracycline under any treatment time was more toxic for S. obliquus than chlortetracycline itself excluding under 24 h. However, the correlation between the toxicity and degradation time was not clear and toxicity enhanced in fact did not follow the increase or decrease in degradation time. Our study showed that the antibiotic chlortetracycline and its degraded products had adverse effect on freshwater phytoplankton, the former has not been reported before and the latter has been overlooked in other research in the past.     

Biodegradation and removal of 3,4-dichloroaniline by Chlorella pyrenoidosa based on liquid chromatography-electrospray ionization-mass spectrometry

3,4-Dichloroaniline (3,4-DCA), widely used in the synthesis of dyes, textile and herbicides, is toxic to living organisms. The purpose of this study was to investigate the capability of green algae in degrading and removing 3,4-DCA in water. An environmentally ubiquitous green alga Chlorella pyrenoidosa was isolated from fresh aquatic environment. Then unicellular alga was incubated with 3,4-DCA at a concentration of 4.6 μg/?mL in water. The residual concentration of 3,4-DCA in the medium and the metabolites were analyzed. A removal percentage of 78.4 % was obtained over a 7-day period. Two major metabolites with less toxicity were identified as 3,4-dichloroformanilide and 3,4-dichloroacetanilide from the liquid chromatography-electrospray ionization-mass spectrometry analysis. The application of microalga C. pyrenoidosa may have potential for removing the environmental pollutant in aquatic environment.     

Enantioselective effects of herbicide imazapyr on Arabidopsis thaliana

The enantioselective toxicity of chiral herbicides in the environment is of increasing concern. To investigate the enantioselective effects of the chiral herbicide imazapyr on target organisms, we exposed Arabidopsis thaliana to imazapyr enantiomers and racemate. The results show that imazapyr was enantioselectively toxic to A. thaliana. The total chlorophyll content in A. thaliana was affected more by (+)-imazapyr than (±)-imazapyr and (?)-imazapyr. Concentrations of proline and malondialdehyde reflected a toxic effect in the order of (+)-imazapyr > (±)-imazapyr > (?)-imazapyr at every concentration. Acetolactate synthase (ALS) activity was inhibited more by (+)-imazapyr than (±)-imazapyr or (?)-imazapyr. At 100 mg L^?1 of imazapyr, ALS activity was 78%, 43%, and 19% with (?)-, (±)-, and (+)-imazapyr, respectively. The results suggest the significant enantioselective toxicity of imazapyr in A. thaliana for greater toxicity with (+)-imazapyr than (±)-imazapyr and (?)-imazapyr, which suggests that (+)-imazapyr has more herbicidal effect.     

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.     

Metribuzin impairs the unicell-colony transformation in the green alga Scenedesmus obliquus

Active growth is a prerequisite for the formation of grazing-protective, mostly eight-celled colonies by the ubiquitous green alga Scenedesmus in response to chemical cues from zooplankton. Colonies can also be evoked by chemically quite similar manmade anionic surfactants, such as FFD-6. In this study, it was hypothesized that growth-inhibiting concentrations of the herbicide metribuzin impair the ability of Scenedesmus obliquus to form colonies in response to the surfactant morphogen FFD-6. The results confirmed that the formation of colonies in S. obliquus was hampered by metribuzin. EC₅₀ values of metribuzin for colony inhibition (approximately 11 μg L⁻¹) were similar to those for growth and photosynthesis inhibition (12-25 μg metribuzin L⁻¹). In the absence of the colony-inducing surfactant FFD-6, S. obliquus populations were comprised of 92% unicells, having on average 1.2 cells per colony at all tested metribuzin concentrations (0-100 μg L⁻¹). In contrast, in the presence of FFD-6 and at low metribuzin concentrations (0 and 5 μg L⁻¹), S. obliquus had more than five cells per colony with a high portion of eight-celled colonies. However, increasing concentrations of metribuzin decreased the number of colonies in the FFD-6-exposed populations and caused them to remain mostly unicellular at the highest concentrations (50 and 100 μg L⁻¹). This study revealed that metribuzin impeded growth and by doing so, also obstructed the possibility for unicellular Scenedesmus to form colonies. Consequently, an increase in mortality of Scenedesmus from grazing is expected.     

Removal and reductive dechlorination of triclosan by Chlorella pyrenoidosa

Triclosan that is widely used as antimicrobial agent has been detected as contaminant in various aquatic environments. In this work, removal and biodegradation of triclosan in water by using a ubiquitous green alga, Chlorella pyrenoidosa was investigated. When C. pyrenoidosa was exposed to a series concentration of triclosan from 100 to 800 ng mL⁻¹, more than 50% of triclosan was eliminated by algal uptake from the culture medium during the first 1 h exposure and reached equilibrium after the 6 h treatment. In the biodegradation experiments, a removal percentage of 77.2% was obtained after C. pyrenoidosa was cultivated with 800 ng mL⁻¹ triclosan for 96 h. A major metabolite from the reductive dechlorination of triclosan was identified by using liquid chromatography coupled with electrospray ionization-mass spectrometry. The ultrastructural morphology of algal cells grown in the presence of triclosan was observed by using transmission electron microscopy and the growth of algal cells was detected. It was found that the trilcosan treatment resulted in the disruption of the chloroplast and the release of organic material into aquatic environment, which indicated that triclosan may affect membrane metabolism.     

In vitro assessment of corticosteroid effects of eight chiral herbicides

Abstract

Information regarding the enantioselective endocrine disruption of chiral herbicides is scarce. This study assessed the disrupting effects of eight typical chiral herbicides on corticosteroids (including glucocorticoids and mineralocorticoids). Enantioselectivity of eight chiral herbicides were evaluated for their agonistic/antagonistic effects on glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) with CHOK1 cell line using reporter gene assay. Their influence on the production of corticosteroids were further investigated in H295R cell line using enzyme-linked immunosorbent assay (ELISA). None of the racemates or enantiomers of eight chiral herbicides exhibited GR or MR agonistic activity at non-cytotoxic concentrations. However, rac-propisochlor and S-imazamox antagonized cortisol-induced transactivation of GR by 21.79% and 38.73% at the concentration of 1.0?×?10^?7 M and 1.0?×?10^?6 M, respectively, and R-napropamide remarkably attenuated aldosterone-induced MR transactivation by 68.78% at 1.0?×?10^?6 M. The secretion of cortisol was significantly restrained after treated with 1.0?×?10^?6 M rac-propisochlor and rac-/R-napropamide at the concentration of 1.0?×?10^?6 M by 26.49%, 30.10% and 35.27%, respectively, while this glucocorticoid was remarkably induced by 1.0?×?10^?5 M rac-diclofop-methyl and its two enantiomers at the concentration of 1.0?×?10^?5 M by 75.60%, 100.1% and 68.78%, respectively. Exposure to rac-propisochlor (1.0?×?10^?6 M), S-diclofop-methyl (1.0?×?10^?5 M) or rac-/S-/R- acetochlor (1.0?×?10^?6 M) and rac-/S-/R-lactofen (1.0?×?10^?6 M) inhibited the secretion of aldosterone by approximately 40%. Our findings suggested that chiral herbicides disrupted corticosteroid homeostasis in an enantioselective way. Therefore, more comprehensive screening is required to better understand the ecological and health risks of chiral pesticides.     

Degradation dynamics,residues and risk assessment of Dufulin enantiomers in corn plants and corn by LC/MS/MS

ABSTRACT

The degradation dynamics and terminal residues of dufulin enantiomers were investigated in two typical corn plants. A convenient and precise chiral method by high-performance liquid chromatography coupled with tandem triple quadrupole mass spectrometry (HPLC/MS/MS) using a Chiralpak IC column was developed and validated for measuring dufulin enantiomers in corn plants and corn. The two enantiomers of dufulin quickly dissipated in the corn plant, and no noticeable stereoselectivity was observed during degradation or in the final residues. After 30% rac-dufulin wettable powder with a 1- to 1.5-fold dose of the recommended value was sprayed two to three times on corn plants, the residue levels of S-(+)-dufulin and R-(-)-dufulin in corn from both sites were lower than or equal to 0.0520 mg kg^–1 on days 7, 14 and 21 after the last application. The dietary risk assessment indicated that dufulin did not exhibit obvious dietary health risks in corn samples when good agricultural practices were implemented. The findings from this study may be used to better understand the chiral profiles of dufulin in the environment and the effect of dufulin residues in corn on health.     

2,4-D Mineralization in soil profiles of a cultivated hummocky landscape in Manitoba,Canada

The objective of this study was to quantify 2,4-D (2,4-dichlorophenoxyacetic acid) mineralization in soil profiles characteristic of hummocky, calcareous-soil landscapes in western Canada. Twenty-five soil cores (8 cm inner diameter, 50 to 125 cm length) were collected along a 360 m transect running west to east in an agricultural field and then segmented by soil-landscape position (upper slopes, mid slopes, lower slopes and depressions) and soil horizon (A, B, and C horizons). In the A horizon, 2,4-D mineralization commenced instantaneously and the mineralization rate followed first-order kinetics. In both the B and C horizons, 2,4-D mineralization only commenced after a lag period of typically 5 to 7 days and the mineralization rate was biphasic. In the A horizon, 2,4-D mineralization parameters including the first-order mineralization rate constant (k ₁), the growth-linked mineralization rate constant (k ₂) and total 2,4-D mineralization at the end of the experiment at 56 days, were most strongly correlated to parameters describing 2,4-D sorption by soil, but were also adequately correlated to soil organic carbon content, soil pH, and carbonate content. In both B and C horizons, there was no significant correlation between 2,4-D mineralization and 2,4-D sorption parameters, and the correlation between soil properties and 2,4-D mineralization parameters was very poor. The k ₁ significantly decreased in sequence of A horizon (0.113% day^?1) > B horizon (0.024% day^?1) = C horizon (0.026% day^?1) and in each soil horizon was greater than k ₂. Total 2,4-D mineralization at 56 days also significantly decreased in sequence of A horizon (42%) > B horizon (31%) = C horizon (27%). In the A horizon, slope position had little influence on k ₁ or k ₂, except that k ₁ was significantly greater in upper slopes (0.170% day^?1) than in lower slopes (0.080% day^?1). Neither k ₁ nor k ₂ was significantly influenced by slope position in the B or C horizons. Total 2,4-D mineralization at 56 days was not influenced by slope positions in any horizon. Our results suggest that, when predicting 2,4-D transport at the field scale, pesticide fate models should consider the strong differences in 2,4-D mineralization between surface and subsurface horizons. This suggests that 2,4-D mineralization is best predicted using a model that has the ability to describe a range of non-linear mineralization curves. We also conclude that the horizontal variations in 2,4-D mineralization at the field scale will be difficult to consider in predictions of 2,4-D transport at the field scale because, within each horizon, 2,4-D mineralization was highly variable across the twenty-five soil cores, and this variability was poorly correlated to soil properties or soil-landscape position.     

Acute toxicity assessment of explosive-contaminated soil extracting solution by luminescent bacteria assays

Explosive-contaminated soil is harmful to people’s health and the local ecosystem. The acute toxicity of its extracting solution was tested by bacterial luminescence assay using three kinds of luminescent bacteria to characterize the toxicity of the soil. An orthogonal test L ₁₆ (4⁵) was designed to optimize the soil extracting conditions. The optimum extracting conditions were obtained when the ultrasonic extraction time, ultrasonic extraction temperature, and the extraction repeat times were 6 h, 40 °C, and three, respectively. Fourier transform infrared spectroscopy (FTIR) results showed that the main components of the contaminated soil’s extracting solution were 2,4-dinitrotoluene-3-sulfonate (2,4-DNT-3-SO₃ ⁻); 2,4-dinitrotoluene-5-sulfonate (2,4-DNT-5-SO₃ ⁻); and 2,6-dinitrotoluene (2,6-DNT). Compared with Photobacterium phosphoreum and Vibrio fischeri, Vibrio qinghaiensis sp. Nov. is more suitable for assessing the soil extracting solution’s acute toxicity. Soil washing can remove most of the contaminants toxic to luminescent bacterium Vibrio qinghaiensis sp. Nov., suggesting that it may be a potential effective remediation method for explosive-contaminated soil.

     

IMPACT OF FOUR PESTICIDES ON THE GROWTH AND METABOLIC ACTIVITIES OF TWO PHOTOSYNTHETIC ALGAE

The acute toxicity was determined for soil algae Chlorella kesslerei and Anabaena inaequalis, exposed to pesticides lindane, pentachlorophenol (PCP), isoproturon (IPU), and methyl parathion (MP). Toxicity markers included growth inhibition, chlorophyll biosynthesis, and total carbohydrate content, as a function of dose and time. Concentration response functions (EC₅₀) were estimated by probit data transformation and weighted linear regression analyses. Lindane's toxicity to Chlorella increased sharply with time (EC₅₀=7490, 10.3, 0.09 mg L^?1; 24, 48, 72 h), but remained nearly constant through 72 h with Anabaena (8.7?6.7 mg L^?1; 24–72 h). PCP at low concentrations stimulated algal growth and chlorophyll a production, an effect reversed at higher doses. Anabaena was less tolerant of PCP and MP than was Chlorella. The 96-h static EC₅₀ values for Chlorella were: 0.003, 34, 0.05, and 291 mg L^?1 for lindane, PCP, isoproturon, and MP, respectively; for Anabaena, these were 4.2, 0.13, 0.21, and 19 mg L^?1. Carbohydrate production responses were similar to those of cell density (growth) and chlorophyll biosynthesis, with MP having the lowest adverse impact. The overall relative toxicity among the four tested pesticides was: for Chlorella, lindane>IPU?PCP?MP; and for Anabaena, PCP>IPU>lindane>MP. The results confirm that toxicants such as these pesticides may affect individual (though related) species to significantly different degrees.     

Time-dependent degradation and toxicity of diclofop-methyl in algal suspensions

Background, aim, and scope  As emerging contaminants, transformation products of the pollutants via various environmental processes are rather unknown, and some may predominately contribute to the environmental risks of the parent compounds. Hence, studies on transformation products complement the assessment of the environmental safety of the parent compounds. In this study, degradation experiments and toxicity tests using diclofop-methyl (DM), a widely used herbicide, and selected major transformation products were carried out in algal cultures to assess the time course of DM toxicity and its relevance in the formation of new breakdown products. Methods  The alga Chlorella vulgaris was maintained in the algal growth medium HB IV. The inhibition of algal growth was determined by measuring optical density at 680 nm (OD₆₈₀). Initially, DM and two selected breakdown products were added to the algal cultures, and following degradation experiments analyses were carried out by high performance liquid chromatography. In addition, the possible relationship between DM degradation and toxicity was assessed, based on physico-chemical properties of the compounds and their toxicity. Results  DM was rapidly absorbed onto the surface of the algal cells where it was hydrolyzed to diclofop (DC). Further degradation to 4-(2, 4-dichlorophenoxy) phenol (DP) occurred in the cells. However, only a minor amount of DC was degraded to DP under the same conditions when DC was initially added to the algal culture. When C. vulgaris was exposed to these compounds for 96 h, the determined EC₅₀ showed that DC was about ten times less toxic than DM (EC₅₀ = 0.42 mg/L) and that DP (EC₅₀ = 0.20 mg/L) was the most toxic. Discussion  Due to strong hydrophobicity and rare dissociation, DM has tendency toward absorption as compared to DC. The higher average degradation rates of DC initially treated by DM revealed the damage of the cell membranes caused by the DM and, thus, enhanced movement of DC into the cells. Following occurrence of phenolic breakdown products, DP suggested that DC should be intracellularly degraded to DP, which had a more potent mode of action and a higher acute toxicity. Moreover, the results for EC₅₀ at various intervals were in accordance with degradation processes of the initial compounds, in which rapid formation of DP was attributed to an increasing toxicity of DM. Conclusions  The toxicity of DM in algal suspensions increased with time due to its degradation to DP, which contributed significantly to the determined toxicity. These results indicate that the toxicity of the pesticide probably depends significantly on degradation. It is thus important to consider the time-dependent environmental processes when evaluating the toxicological effects of pesticides for proper risk assessment. Recommendations and perspectives  Increasing transformation products of these contaminants are identified in the environment, although they seem to be unknown in terms of the lacking studies on environmental behavior and ecotoxicity concerning them. Certain breakdown products probably greatly contribute to the apparent toxicity of the parent compounds, which is ascribed to the parent compounds in general studies ignoring the dependence of their toxicity on various transformation pathways. These studies that identify new intermediates and assess their toxicity via the environmental processes will be helpful to distinguish the nature of toxicity of the parent contaminants.