Acute, chronic and sublethal effects of the herbicide propanil on Daphnia magna

Acute and chronic toxicity tests with propanil were conducted on Daphnia magna. The 24 and 48 h LC50 were 43.74 and 5.01 mg/l respectively. Chronic toxicity tests were carried out using sublethal propanil concentrations (0.07, 0.10, 0.21 and 0.55 mg/l) during 21 days. The effect of propanil on survival, reproduction and growth of D. magna organisms was monitored. The parameters used to evaluate herbicide effect on reproduction were: mean total young ones per female, mean brood size, time to first reproduction, mean number broods per female and intrinsic rate of natural increase (r). Survival and growth (body length) were also determined after 21 days of exposure to the herbicide. Reproduction was significantly reduced when propanil concentration increased in the medium. The intrinsic rate of natural increase (r) decreased with increasing concentrations of propanil especially in those animals exposed to 0.55 mg/l. However, growth as well as survival of the exposed organisms only decreased in daphnids exposed to the highest propanil concentration tested. The maximum acceptable toxicant concentration (MATC) was calculated for D. magna exposed to the herbicide using as parameter of evaluation the intrinsic rate of natural increase (r). The interpolation of these results gave MATC values of 0.08 mg/l herbicide. We have derived the EC50 values for some selected parameters on D. magna exposed to propanil. EC50 values indicated that reproductive parameters were very sensitive of the effect of propanil on daphnids. Finally, the daphnids were exposed to the same sublethal herbicide concentrations as in the chronic study and the effect of the toxicant on filtration and ingestion rates was determined. Feeding rates of D. magna declined with increasing propanil concentrations. The effective propanil concentrations at which feeding rates were reduced to 50% of that in controls (EC50) were also calculated.     

Evaluation of a Daphnia magna renewal life‐cycle test method with diazinon

Abstract

Acute and chronic toxicity tests with diazinon (diethyl 2‐isopropyl‐6‐methyl‐4‐pyrimidinyl phosphorothionate) were conducted on Daphnia magna. The 24‐hr static LC50 was 0.86 μL.L^‐1. The sublethal effects of 0.05, 0.1, 0.5, 0.75 and 1.0 ngL^‐1 of diazinon concentrations on the survival, reproduction and growth of D. magna were monitored for 21 days. The algae Nannochloris oculata (5 x 10⁵ cellsmL^‐1) was used to feed the daphnids. The parameters used to determined the effect of the pesticide on D. magna were: mean total young per female; mean brood size; days to first brood; intrinsic rate of natural increase (r); growth; and survival. Reproduction as well as survival was significantly reduced at diazinon concentrations of 0.10 ngL^‐1 and higher. The intrinsic rate of natural increase (r) decreased with increasing concentrations of diazinon. Growth, as measured by body length, was depressed significantly at 0.05 ngL^‐1 of diazinon and higher concentrations. The maximum acceptable toxicant concentration (MATC) was calculated. The chronic data was used to formulate an acute/chronic ratio.     

Population dynamics in Daphnia magna as modified by chronic tetradifon stress

Two Daphnia magna offsprings (animals from the first and third brood) whose parentals (F0-generation) were exposed during 21 days to different tetradifon (4-chlorophenyl 2,4,5-trichlophenyl sulfone) concentrations, were transferred to a pesticide free medium during 21 days (recovery period). The algae Nannochloris oculata (5 x 10(5) cells/mL) was used as food. In this recovery study, survival, growth and reproduction (mean total young per female, mean brood size, onset of reproduction and mean number broods per female) were assessed as individual parameters and the intrinsic rate of natural increase (r) as population parameter, for F1 generation (1st and 3rd broods). Reproduction was still reduced in F1 (1st and 3rd broods) generation daphnids from parentals (F0) exposed to 0.18 mg/L tetradifon and higher concentrations even after 21 days in clean water. However, survival was not significantly different (p > 0.05) in those F1 offsprings from parentals pre-exposed to the acaricide. Growth was still reduced in F1 daphnids from parentals pre-exposed to 0.10-0.44 mg/L tetradifon. The intrinsic rate of natural increase (r) was still affected in F1 generation daphnids, specially in those from the third brood. Therefore, F1 generation D. magna offsprings from a parental generation (F0) previously exposed to the acaricide tetradifon were not able to recuperate completely when a recovery period of 21 days was allowed.     

Influence of sediment on the fate and toxicity of a polyethoxylated tallowamine surfactant system (MON 0818) in aquatic microcosms

The fate and toxicity of a polyethoxylated tallowamine (POEA) surfactant system, MON 0818, was evaluated in water-sediment microcosms during a 4-d laboratory study. A surfactant solution of 8 mg l(-1) nominal concentration was added to each of nine 72-l aquaria with or without a 3-cm layer of one of two natural sediments (total organic carbon (TOC) 1.5% or 3.0%). Control well water was added to each of nine additional 72-l aquaria with or without sediment. Water samples were collected from the microcosms after 2, 6, 24, 48, 72, and 96 h of aging to conduct 48-h toxicity tests with Daphnia magna and to determine surfactant concentrations. Elevated mortality of D. magna (43-83%) was observed in overlying water sampled from water-only microcosms throughout the 96-h aging period, whereas elevated mortality (23-97%) was only observed in overlying water sampled from water-sediment microcosms during the first 24h of aging. Measured concentrations of MON 0818 in water-only microcosms remained relatively constant (4-6 mg l(-1)) during the 96-h period, whereas the concentrations in overlying water from microcosms containing either of the two types of sediment dissipated rapidly, with half-lives of 13 h in the 3.0% TOC sediment and 18 h in the 1.5% TOC sediment. Both toxicity and the concentration of MON 0818 in overlying water decreased more rapidly in microcosms containing sediment with the higher percent TOC and clay and with a higher microbial biomass. Mortality of D. magna was significantly correlated with surfactant concentrations in the overlying water. These results indicate that the toxicity of the POEA surfactant in water rapidly declines in the presence of sediment due to a reduction in the surfactant concentration in the overlying water above the sediment.     

Thiobencarb toxicity and plasma AChE inhibition in the European eel

The acute toxicity of the herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate) was determined for the European eel (Anguilla anguilla). The 24, 48, 72 and 96 hours median lethal concentrations (LC50) were 25.7, 21.7, 17.0 and 13.2 mg/L, respectively. Fish were also exposed to a sublethal thiobencarb concentration (1/60 LC50-96 hr = 0.22 mg/L) during 96 hours in a flow-through system and then an elimination period of 192 hours in clean water was allowed. Eels were removed and blood samples taken out at each exposure time and recovery period in order to evaluate AChE activity. Thiobencarb induced significant inhibitory effects on plasma AChE activity of A. anguilla from the first contact time with the toxicant. This inhibition (under 50% activity) was maintained during the entire exposure period (96 hours) and even those animals transferred to clean water showed plasma AChE activity different from the controls. Differences between total and specific AChE activity were detected during the exposure period. Total AChE activity in the plasma from animals transferred to a medium free of toxicant recovered its normal value while specific AChE activity remained depressed (< 50%) until five days later.     

Effects of the anti-fouling herbicide Irgarol 1051 on two life stages of the grass shrimp,Palaemonetes pugio

This study investigated lethal and sublethal effects (glutathione, lipid peroxidation, cholesterol, and acetylcholinesterase) of the anti-fouling herbicide Irgarol 1051 on larval and adult grass shrimp (Palaemonetes pugio). The 96-hour LC50 test for larvae resulted in an estimated LC50 of 1.52 mg/L (95% confidence interval [CI] 1.26–1.85 mg/L). The adult 96-h LC50 was 2.46 mg/L (95% CI = 2.07–2.93 mg/L). Glutathione, lipid peroxidation, cholesterol and acetylcholinesterase levels were not significantly affected in adult grass shrimp by exposure of up to 3.00 mg/L irgarol. Lipid peroxidation and acetylcholinesterase levels in the larvae were significantly higher than controls in the highest irgarol exposures of 1.0 and 2.0 mg/L, respectively. Cholesterol levels were significantly reduced in larvae in all four irgarol concentrations tested while glutathione levels were not significantly affected in larvae. Both lethal and sublethal effects associated with irgarol exposure were only observed at concentrations well above those reported in the environment.     

Effects of the anti-fouling herbicide Irgarol 1051 on two life stages of the grass shrimp, Palaemonetes pugio

This study investigated lethal and sublethal effects (glutathione, lipid peroxidation, cholesterol, and acetylcholinesterase) of the anti-fouling herbicide Irgarol 1051 on larval and adult grass shrimp (Palaemonetes pugio). The 96-hour LC50 test for larvae resulted in an estimated LC50 of 1.52 mg/L (95% confidence interval [CI] 1.26-1.85 mg/L). The adult 96-h LC50 was 2.46 mg/L (95% CI = 2.07-2.93 mg/L). Glutathione, lipid peroxidation, cholesterol and acetylcholinesterase levels were not significantly affected in adult grass shrimp by exposure of up to 3.00 mg/L irgarol. Lipid peroxidation and acetylcholinesterase levels in the larvae were significantly higher than controls in the highest irgarol exposures of 1.0 and 2.0 mg/L, respectively. Cholesterol levels were significantly reduced in larvae in all four irgarol concentrations tested while glutathione levels were not significantly affected in larvae. Both lethal and sublethal effects associated with irgarol exposure were only observed at concentrations well above those reported in the environment.     

Phytotoxicity and antioxidative enzymes of green microalga (Desmodesmus subspicatus) and duckweed (Lemna minor) exposed to herbicides MCPA, chloridazon and their mixtures

In this study, we evaluate the toxicity of MCPA (auxin-like growth inhibitor), chloridazon (CHD) (PSII-inhibitor) and their mixtures to floating plants and planktonic algae. Toxicity of MCPA (4-chloro-2-methylphenoxyacetic acid) and CHD (5-amino-4-chloro-2-phenyl-3(2H)-pyridazinone) was first assessed in two growth inhibition tests with Lemna minor (ISO/DIS 20079) and Desmodesmus subspicatus (ISO 8692). Next, herbicide mixtures at concentrations corresponding to the EC values were used to assess their interactive effects, and the biomarkers were: for duckweed fresh weight, frond area, chlorophyll content and number of fronds, and for algae cell count and cell volume. The 3d EC?? and EC?? values using cell counts of D. subspicatus were 142.7 and 529.1 mg/L for MCPA and 1.7 and 5.1 mg/L for CHD. The 7d EC?? and EC?? values using frond number of L. minor amounted to 0.8 and 5.4 mg/L for MCPA and 0.7 and 10.4 mg/L for CHD. Higher sensitivity of reproductive (number of cells/fronds) than growth processes (cell volume/frond area) to herbicides applied individually and in mixtures was especially pronounced in the responses of Desmodesmus. Herbicide interactions were assessed by the two-way ANOVA and Abbott's formula. Generally, an antagonistic interaction with Lemna was revealed by MCPA and chloridazon, whereas additive effect of both herbicides was observed for Desmodesmus. A significant stimulation of SOD and APX activity by binary mixtures was noted in algal cells mainly after 24 and 48 hours of exposure. The extremely high stimulation of the activity of both enzymes was induced by the combination EC??CHD + EC??MCPA (48 h). Presumably due to oxidative stress, the treatment with CHD at concentration EC?? after 72 h was lethal for algae grown in aerated cultures, in contrast to standardized test conditions. Taking into account the consequences of risk assessment for herbicide mixtures we can state that a relatively low toxicity, as well as the lack of significant synergy between MCPA and CHD to non-target plants appears to be the most important result.     

Investigation of acute toxicity and the effect of 2,4-D (2,4-dichlorophenoxyacetic acid) herbicide on the behavior of the common carp (Cyprinus carpio L., 1758; Pisces,Cyprinidae)

A 96-h LC(50) values of 2,4-D [(2,4-dichlorophenoxy)acetic acid], a common contaminating agricultural herbicide, were determined on the adult common carp (Cyprinus carpio L., 1758; Pisces, Cyprinidae). The study was conducted in two stages using 130 carp. The data obtained were statistically evaluated by the use of the EPA computer program based on Finney's Probit Analysis Method and a 96-h LC(50) value for C. carpio L., 1758 was found to be 63.24 mg/l in a static bioassay test system. 95% lower and upper confidence limits for the LC(50) were 55.03 and 71.92 mg/l, respectively. Water temperature was 17+/-1 degrees C. Behavioral changes of the above mentioned species were examined for various herbicide concentrations.     

Effect of pulse frequency and interval on the toxicity of chlorpyrifos to Daphnia magna.

Due to the episodic nature in which organisms are exposed to non-point source pollutants, it is necessary to understand how they are affected by pulsed concentrations of contaminants. This is essential, as standard toxicity tests may not adequately simulate exposure scenarios for short-lived hydrophobic compounds, such as chlorpyrifos (CPF), a broad-spectrum organophosphate insecticide. Studies were conducted with 7-day old Daphnia magna for 7 days to evaluate the effect of pulse frequency and interval among multiple CPF exposures. Daphnids were exposed to a total exposure of either 12 h at 0.5 microg/l or 6 h at 1.0 microg/l nominal CPF, respectively, in all studies. For interval studies, D. magna were exposed to two pulses of CPF at each concentration, with 0-96-h intervals between pulses. For frequency studies, D. magna were exposed to each CPF concentration altering the pulse scheme by decreasing the exposure duration but increasing the number of pulses, keeping the total exposure time the same. The pulse interval between multiple pulses in these experiments was 24 h. Our results suggest that D. magna can withstand an acutely lethal CPF exposure provided that there is adequate time for recovery between exposures.     

Aquatic toxicity of cartap and cypermethrin to different life stages of Daphnia magna and Oryzias latipes

Cartap and cypermethrin, which are among the most widely used pesticides in many countries, are considered safe because of their low mammalian toxicity and their low persistence in the environment. However, recent findings of endocrine-disrupting effects and developmental neurotoxicity have raised concerns about the potential ecological impacts of these pesticides. We evaluated the aquatic toxicity of cartap [S,S'-(2-dimethylaminotrimethylene) bis(thiocarbamate), unspecified hydrochloride] and cypermethrin [(RS)-alpha-cyano-3-phenoxybenzyl-(1RS,3RS,1RS,3SR)-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylate], both individually and combined, on different life stages of the freshwater cladoceran Daphnia magna and a freshwater teleost, Japanese medaka (Oryzias latipes). The 96-hr Daphnia median effective concentrations (EC50s) for cartap and cypermethrin were 91.0 microg/L and 0.00061 microg/L, respectively. Rapid recovery of Daphnia was observed after short-term pulsed exposure to cartap and cypermethrin; there were no adverse effects on reproduction or survival 20 d after a 24 hr exposure to cartap up to 1240 microg/L and cypermethrin up to 1.9 microg/L. Chronic continuous exposure (for 21 d) of 7-d-old Daphnia to cypermethrin significantly reduced the intrinsic population growth rate in a concentration-dependent manner. However, because the intrinsic population growth rates were all above zero, populations did not decrease even at the highest experimental concentration of 200 ng/L. Exposure of Daphnia neonates (< 24 hr old) to cypermethrin for 21 d caused significant, sub-lethal reproduction-related problems, such as increased time to first brood, reduced brood size, and reduced total brood number, at 0.0002, 0.002, and 0.2 ng/L cypermethrin, but the intrinsic population growth rate was not significantly affected. Oryzias latipes was relatively more resistant to both pesticides. In particular, embryos appeared to be more resistant than juveniles or adults, which may be partly due to the protective role of the chorion. The incidence of larval fish deformity was significantly higher after a 96 hr exposure to as low as 250 microg/L of cartap or 40 microg/L of cypermethrin. The mixture of both compounds showed no synergistic toxicity. The extremely high acute-to-chronic ratio suggests that the standard acute lethal toxicity assessment might not reflect the true environmental hazards of these frequently used pesticides. Ecological hazard assessments of long-term low dose or pulsed exposures to cartap and cypermethrin may reveal more realistic consequences of these compounds in surface water.     

Phytotoxicity and antioxidative enzymes of green microalga (Desmodesmus subspicatus) and duckweed (Lemna minor) exposed to herbicides MCPA,chloridazon and their mixtures

In this study, we evaluate the toxicity of MCPA (auxin-like growth inhibitor), chloridazon (CHD) (PSII-inhibitor) and their mixtures to floating plants and planktonic algae. Toxicity of MCPA (4-chloro-2-methylphenoxyacetic acid) and CHD (5-amino-4-chloro-2-phenyl-3(2H)-pyridazinone) was first assessed in two growth inhibition tests with Lemna minor (ISO/DIS 20079) and Desmodesmus subspicatus (ISO 8692). Next, herbicide mixtures at concentrations corresponding to the EC values were used to assess their interactive effects, and the biomarkers were: for duckweed fresh weight, frond area, chlorophyll content and number of fronds, and for algae cell count and cell volume. The 3d EC₁₀ and EC₅₀ values using cell counts of D. subspicatus were 142.7 and 529.1 mg/L for MCPA and 1.7 and 5.1 mg/L for CHD. The 7d EC₁₀ and EC₅₀ values using frond number of L. minor amounted to 0.8 and 5.4 mg/L for MCPA and 0.7 and 10.4 mg/L for CHD. Higher sensitivity of reproductive (number of cells/fronds) than growth processes (cell volume/frond area) to herbicides applied individually and in mixtures was especially pronounced in the responses of Desmodesmus. Herbicide interactions were assessed by the two-way ANOVA and Abbott's formula. Generally, an antagonistic interaction with Lemna was revealed by MCPA and chloridazon, whereas additive effect of both herbicides was observed for Desmodesmus. A significant stimulation of SOD and APX activity by binary mixtures was noted in algal cells mainly after 24 and 48 hours of exposure. The extremely high stimulation of the activity of both enzymes was induced by the combination EC₁₀CHD + EC₅₀MCPA (48 h). Presumably due to oxidative stress, the treatment with CHD at concentration EC₅₀ after 72h was lethal for algae grown in aerated cultures, in contrast to standardized test conditions. Taking into account the consequences of risk assessment for herbicide mixtures we can state that a relatively low toxicity, as well as the lack of significant synergy between MCPA and CHD to non-target plants appears to be the most important result.     

Selected endocrine disrupting compounds (Vinclozolin,Flutamide, Ketoconazole and Dicofol): Effects on survival,occurrence of males,growth, molting and reproduction of <Emphasis Type="Italic">Daphnia magna</Emphasis>

BACKGROUND, AIM AND SCOPE: Pollution-induced endocrine disruption in vertebrates and invertebrates is a worldwide environmental problem, but relatively little is known about effects of endocrine disrupting compounds (EDCs) in planktonic crustaceans (including Daphnia magna). Aims of the present study were to investigate acute 48 h toxicity and sub-chronic (4-6 days) and chronic (21 days) effects of selected EDCs in D. magna. We have investigated both traditional endpoints as well as other parameters such as sex determination, maturation, molting or embryogenesis in order to evaluate the sensitivity and possible use of these endpoints in ecological risk assessment. MATERIALS AND METHODS: We have studied effects of four model EDCs (vinclozolin, flutamide, ketoconazole and dicofol) on D. magna using (i) an acute 48 h immobilization assay, (ii) a sub-chronic, 4-6 day assay evaluating development and the sex ratio of neonates, and (iii) a chronic, 21 day assay studying number of neonates, sex of neonates, molting frequency, day of maturation and the growth of maternal organisms. RESULTS: Acute EC50 values in the 48 h immobilization test were as follows (mg/L): dicofol 0.2, ketoconazole 1.5, flutamide 2.7, vinclozolin >3. Short-term, 4-6 day assays with sublethal concentrations showed that the sex ratio in Daphnia was modulated by vinclozolin (decreased number of neonate males at 1 mg/L) and dicofol (increase in males at 0.1 mg/L). Flutamide (up to 1 mg/L) had no effect on the sex of neonates, but inhibited embryonic development at certain stages during chronic assay, resulting in abortions. Ketoconazole had no significant effects on the studied processes up to 1 mg/L. DISCUSSION: Sex ratio modulations by some chemicals (vinclozolin and dicofol) corresponded to the known action of these compounds in vertebrates (i.e. anti-androgenicity and anti-oestrogenicity, respectively). Our study revealed that some chemicals known to affect steroid-regulated processes in vertebrates can also affect sublethal endpoints (e.g. embryonic sex determination and/or reproduction) in invertebrates such as D. magna. CONCLUSIONS: A series of model vertebrate endocrine disrupters affected various sub-chronic and chronic parameters in D. magna including several endpoints that have not been previously studied in detail (such as sex determination in neonates, embryogenesis, molting and maturation). Evaluations of traditional reproduction parameters (obtained from the 21 day chronic assay). as well as the results from a rapid, 4-6 day, sub-chronic assay provide complementary information on non-lethal effects of suspected organic endocrine disrupters. RECOMMENDATIONS AND PERSPECTIVES: It seems that there are analogies between vertebrates and invertebrates in toxicity mechanisms and in vivo effects of endocrine disruptors. However, general physiological status of organisms may also indirectly affect endpoints that are traditionally considered 'hormone regulated' (especially at higher effective concentrations as observed in this study) and these factors should be carefully considered. Further research of D. magna physiology and comparative studies with various EDCs will help to understand mechanisms of action as well as ecological risks of EDCs in the environment.     

Mixture toxicity of the antifouling compound irgarol to the marine phytoplankton species Dunaliella tertiolecta

This study examined the toxicity of irgarol, individually and in binary mixtures with three other pesticides (the fungicide chlorothalonil, and the herbicides atrazine and 2,4-D), to the marine phytoplankton species Dunaliella tertiolecta. Standard 96-h static algal bioassays were used to determine pesticide effects on population growth rate. Irgarol significantly inhibited D. tertiolecta growth rate at concentrations > or = 0.27 micro g/L. Irgarol was significantly more toxic to D. tertiolecta than the other pesticides tested (irgarol 96 h EC50 = 0.7 micro g/L; chlorothalonil 96 h EC50 = 64 micro g/L; atrazine 96 h EC50 = 69 micro g/L; 2,4-D 96 h EC50 = 45,000 micro g/L). Irgarol in mixture with chlorothalonil exhibited synergistic toxicity to D. tertiolecta, with the mixture being approximately 1.5 times more toxic than the individual compounds. Irgarol and atrazine, both triazine herbicides, were additive in mixture. The toxicity threshold of 2,4-D was much greater than typical environmental levels and would not be expected to influence irgarol toxicity. Based on these interactions, overlap of certain pesticide applications in the coastal zone may increase the toxicological risk to resident phytoplankton populations.     

Effects of lindane on Daphnia magna during chronic exposure

Abstract

Acute and chronic toxicity tests with lindane were conducted on Daphnia magna. The 24‐hr static LC50 was 1.64 mgL^‐1. The sublethal effects of 0.16, 0.25, 0.32, 0.60 and 0.80 mgL^‐1 lindane on the survival, reproduction and growth of D. magna were monitored for 21 days. The algae Nannochloris oculata (5 × 10⁵ cellsmL^‐1) was used to feed the daphnids. The parameters used to determined the effect of the pesticide on D. magna were:mean total young per female, mean brood size, days to first brood, intrinsic rate of natural increase (r), growth, and survival. Reproduction as well as survival was significantly reduced at lindane concentrations of 0.25 mgL^‐1 and higher. The intrinsic rate of natural increase (r) decreased with increasing concentrations of lindane. Growth, as measured by body length, was depressed significantly at 0.25 mgL^‐1 lindane and higher. The chronic data was used to formulate an acute/chronic ratio.     

A freshwater shrimp (Paratya compressa improvisa) as a sensitive test organism to pesticides

The susceptibility of 2-week-old individuals of the freshwater shrimp, Paratya compressa improvisa, to five kinds of insecticide and five kinds of herbicide was examined in comparison with that of two species of Cladocera, Daphnia magna and Moina macrocopa. The shrimp was especially sensitive to two organophosphorus insecticides. The 48-h LC50 values for fenitrothion and fenthion to the shrimp were 1.15 and 1.04 microg litre(-1) (mean value, n=2), in contrast with 37.8 and 35.3 microg litre(-1) in the case of M. macrocopa, and more than 50 microg litre(-1) with D. magna. The shrimp also showed the higher susceptibility to other insecticides, diazinon, carbaryl (NAC) and BPMC, apart from D. magna to diazinon and NAC. The shrimp also showed higher susceptibility to herbicides. The 48-h LC50 values of CNP, benthiocarb, oxadiazon, butachlor, and symetryne to the shrimp were two to eight times lower than those of two species of Cladocera, except for the LC50 value of oxadiazon to M. macrocopa, which was very slightly higher. However, the shrimp showed a somewhat lower susceptibility to heavy metals than the two species of Cladocera, especially to copper, and to cadmium and zinc in comparison with D. magna. A bioassay using the shrimp with river water, collected from the river adjacent to the paddy field, showed clearly the high mortality of the shrimp following the aerial spraying with pesticides.     

METIER (Modular Ecotoxicity Tests Incorporating Ecological Relevance) for difficult substances--III. Effects of medium renewal and use of a carrier on the bioavailability of parathion

In this work we compared the effects of medium renewal and use of a carrier (ethanol) in the lethal toxicity of parathion to Daphnia magna Straus. Actual concentrations measured in test solutions with and without ethanol were not significantly different, indicating that there is no need for a carrier solution to assist dissolution of parathion, up to a concentration of 5 mg litre(-1). LC50 values at 48 h from bioassays with and without medium renewal were identical, indicating that in our experimental conditions, degradation of parathion was not a significant consideration. Similar 48-h LC50 values were obtained in all tests, with or without ethanol present. Thus, potential interactions between carrier and toxicant had no effect on the lethal toxicity of parathion to D. magna.     

Chronic toxicity tests with Daphnia magna for examination of river water quality

Chronic toxicity tests with Daphnia magna were applied for examination of river water quality. Water was sampled from the Maioka River in Yokohama City on May 14, 20, and 27, 1999, and used for the test after solid-phase extraction. The chronic test was carried out according to the OECD method. The duration was 21 days and the total number of live offspring produced per parent animal was counted. The results of the tests showed, survival rates of 100% using river water sampled on May 14 and 20 and the total numbers of live offspring produced per parent animal did not differ from the control. However, the survival rate of the sample collected on May 27 was 0% and the pesticides, fenitrothion, and thiobencarb were detected in the water. In addition to the river water samples, reconstituted water (Elendt M7) with additions of fenitrothion and thiobencarb was prepared to investigate mortality. When the reconstituted water with thiobencarb was applied to the test, the total number of live offspring produced per parent animal did not differ from the control. In contrast, when reconstituted water with fenitrothion was applied to the test, most parents were alive, but the total number of live offspring produced per parent animal was apparently different. The results of the above tests indicate that D. magna was affected not only by fenitrothion in the river water collected on May 27, but also by other factors that were not clarified in this study.     

Assessment of silver nanoparticle-induced physiological and molecular changes in Arabidopsis thaliana

In this study, the effect of silver nanoparticles and silver ions on Arabidopsis thaliana was investigated at physiological and molecular levels. The seedlings were grown in sublethal concentrations of silver nanoparticles and silver ions (0.2, 0.5, and 1 mg/L) in 1/4 Hoagland’s medium for 14 days under submerged hydroponic conditions. Significantly higher reduction in the total chlorophyll and increase in anthocyanin content were observed after exposure to 0.5 and 1 mg/L silver nanoparticles as compared to similar concentrations of silver ions. Lipid peroxidation increased significantly after exposure to 0.2, 0.5, and 1 mg/L of silver nanoparticles and 0.5 and 1 mg/L of silver ions. Qualitative analysis with dichloro-dihydro-fluorescein diacetate and rhodamine 123 fluorescence showed a dose-dependent increase in reactive oxygen species production and changes in mitochondrial membrane potential in the roots of seedlings exposed to different concentrations of silver nanoparticles. Real-time PCR analysis showed significant upregulation in the expression of sulfur assimilation, glutathione biosynthesis, glutathione S-transferase, and glutathione reductase genes upon exposure to silver nanoparticles as compared with silver ions. Overall, based on the physiological and molecular level responses, it was observed that exposure to silver nanoparticles exerted more toxic response than silver ions in A. thaliana.     

Effects of nickel chloride and oxygen depletion on behaviour and vitality of zebrafish (Danio rerio, Hamilton, 1822) (Pisces, Cypriniformes) embryos and larvae

We examined acute (2 h exposure of 5-day-old larvae) and subchronic (exposure from fertilization up to an age of 11 days) effects of NiCl(2).6H2O on embryos and larvae of zebrafish (Danio rerio), both alone and in combination with oxygen depletion. The following endpoints were recorded: acute exposure: locomotory activity and survival; subchronic exposure: hatching rate, deformations, locomotory activity (at 5, 8 and 11 days) and mortality. In acute exposures nickel chloride (7.5-15 mg Ni/L) caused decreasing locomotory activity. Oxygen depletion (or=10 mg Ni/L resulted in delayed hatching at an age of 96 h, in decreased locomotory activity at an age of 5 days, and increased mortality at an age of 11 days (LC20=9.5 mg Ni/L). The observed LOEC for locomotory activity (7.5 mg Ni/L) is in the range of environmentally relevant concentrations. Since locomotory activity was already affected by acute exposure, this parameter is recommended to supplement commonly recorded endpoints of toxicity.