Cyst-based toxicity tests X: comparison of the sensitivity of the acute Daphnia magna test and two crustacean microbiotests for chemicals and wastes

This paper statistically evaluates the sensitivity of the acute Daphnia magna bioassay in comparison to that of two crustacean microbiotests: the Streptoxkit F and Thamnotoxkit F, which make use of larvae of the anostracans Streptocephalus proboscideus and Thamnocephalus platyurus respectively, hatched from cysts. Regression equations were calculated for 146 data pairs, taken from 5 different studies dealing with the acute toxicity of pure chemicals, effluents, river sediments, solid wastes and monitoring wells, and sludges. All comparisons show that there is a significant relationship (p< 0.05) between the acute effects found with the D.magna and the two crustacean microbiotests; correlation coefficients ranged from 0.84 to 0.92.

In the majority of cases, effect ratios between the conventional D.magna and the microbiotests were within a factor 2 for both pure chemicals and environmental samples.

Variation coefficients for repeated tests on the reference chemical potassium dichromate indicate a good precision and hence a good degree of standardization of the microbiotest procedures. The evidence provided in this paper demonstrates that the new crustacean microbiotests can be used as low cost alternatives to the conventional D.magna acute assay.     

How accurately do semi-permeable membrane devices measure the bioavailability of polycyclic aromatic hydrocarbons to Daphnia magna?

Semi-permeable membrane devices (SPMDs) are passive samplers that have been designed to sample the bioavailable fractions of hydrophobic organic compounds in aquatic ecosystems. This study aims at evaluating the ability for SPMD to sample polycyclic aromatic hydrocarbons (fluoranthene, pyrene and benzo[a]pyrene) that are actually bioavailable to Daphnia magna. For that purpose, the SPMD-available fraction and the bioavailable fraction to D. magna are compared in controlled media with Dissolved Organic Matters (DOMs) from various origins and at different concentrations. The presence of all but one DOM reduces the accumulation of PAHs in SPMD or in D. magna. Moreover, this comparative laboratory study shows that in 10 cases on 13, the SPMD-available fraction is close to the available fraction to D. magna. When significant differences are observed between SPMD-available and bioavailable fractions, they remain less than 50% at DOM concentrations below 10 mg/l DOC, which corresponds to a maximum DOC concentration usually found in temperate rivers. This study confirms the suitability of the SPMD technique to monitor readily bioavailable hydrophobic contaminants in aquatic environments containing DOM from various origins and characteristics.     

Toxicity of HC Orange No. 1 to Daphnia magna, zebrafish (Brachydanio rerio) embryos, and goldfish (Carassius auratus)

HC Orange No. 1 (HCO1; 2-nitro-4′-hydroxydiphenylamine) (CAS No. 54381-08-7) is used as a color additive in hair dyes and can be released into aquatic environments in wastewater. In this paper, the effects of HCO1 on aquatic organisms were studied using a battery of toxicological tests. These included measuring immobilization of Daphnia magna, inhibition of zebrafish embryo development, and acute lethality in zebrafish and goldfish, which are different species belonging to different trophic levels. HCO1 was toxic to all of the organisms studied. In our experiments, HCO1 remarkably restrained the mobility of D. magna, which may cause subsequent death. The EC₅₀ value for restrained the mobility of D. magna at 48 h was 1.54 mg HCO1 l⁻¹. In addition, HCO1 showed toxicity in zebrafish and goldfish, where LC₅₀values at 96 h were 4.04 and 5.37 mg l⁻¹, respectively. The results also indicated that HCO1 remarkably retarded the development of zebrafish embryos, which may cause embryo abnormality and even lethality. The most sensitive toxicological endpoint in the development of the embryos was failure to hatch, which had an EC₅₀ of 0.19 mg HCO1 l⁻¹. These results indicated that HCO1 is a potential teratogen to zebrafish embryos. In addition, as HCO1 concentrations increased, the outcomes of each of these toxicity tests changed in a concentration-dependent manner. Together, the results revealed that HCO1 appears to be toxic to multiple different species of aquatic organisms. The EC₅₀ (LC₅₀) values contain sufficient discriminatory power for risk assessment of HCO1 in aquatic environments. Based on the present results, more efficient risk assessment procedures for HCO1 will be designed in the future, integrating more flexible testing methods into the testing schemes that employ only the necessary tools for each case.     

Culture techniques for three freshwater macroinvertebrate species and their use in toxicity tests

At present only one freshwater invertebrate species (Daphnia magna) is widely used in routine toxicity tests in the United Kingdom. In this paper methods are described for the continuous laboratory culture of three European macroinvertebrates species, the dipteran Chironomus riparius, the amphipod Gammarus pulex and the isopod Asellus aquaticus. Their use as test species to assess both acute and chronic toxicity responses is discussed.     

The copper tolerance strategies and the role of antioxidative enzymes in three plant species grown on copper mine

This study was undertaken to identify the strategies and the status of antioxidant enzyme activities involved in three plant species tolerance against Cu-toxicity in copper mine. The following methods were used for evaluations in three wild type species; Datura stramonium, Malva sylvestris and Chenopodium ambrosioides. The level of chlorophyll and the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) by spectrometry, malondialdehyde (MDA) and dityrosine by HPLC and the levels of Cu in tissues and soils by atomic absorption spectrometry (AAS).

Analysis showed that total and available copper were at toxic levels for plants growing on contaminated soil (zone 1). However, there were not any visual and conspicuous symptoms of Cu toxicity in plant species. Among three species, excess copper was transferred only into the D. stramonium and C. ambrosioides tissues. The C. ambrosioides accumulated Cu in roots and then in leaves, in which the leaves chloroplasts stored Cu around two times of vacuoles. In D. stramonium most of Cu was accumulated in leaves in which the storage rate in vacuoles and chloroplasts were 42% and 8%, respectively. In zone 1, the chlorophyll levels increased significantly in leaves of C. ambrosioides with respect to the same plant growing on uncontaminated soil (zone 2). There was insignificant decrease in chlorophyll content of D. stramonium leaves, collected from zone 1 with respect to zone 2. The D. stramonium and C. ambrosioides in zone 1, both revealed significant increase in their tissues antioxidant enzyme activities in comparison with the same samples of zone 2. There was significant elevation in oxidative damage biomarkers; MDA and dityrosine, when the aerial parts of D. stramonium in zone 1 were compared with the same parts of zone 2.

We concluded that there were different tolerance strategies in studied plant species that protected them against copper toxicity. In M. sylvestris, exclusion of Cu from the roots or its stabilization in the soil restricted Cu toxicity effects. On the other hand D. stramonium and C. ambrosioides, elevated their antioxidative enzyme activities in response to cu-toxicity. In addition, the species D. stramonium accumulated excess of Cu in leaves vacuoles.     

Environmental fate of synthetic pyrethroids during spray drift and field runoff treatments in aquatic microcosms.

The aquatic fate and persistence of synthetic pyrethroids under spray drift and field runoff treatment regimens were determined in outdoor pond microcosms. In this paper, the experimental design and construction of outdoor microcosms is presented, as well as the aquatic fate of tralomethrin and deltamethrin. Tralomethrin is rapidly degraded to deltamethrin, with a half-life of 12.7 hours under spray drift conditions. Degradation profiles of tralomethrin in water indicated rapid conversion of deltamethrin and to less active isomers and then to decamethrinic acid (BR2CA). After 24 hours, the percent radioactivity of tralomethrin was 25% of the test material in the water column. In sediment, tralomethrin was immediately converted to deltamethrin. Deltamethrin is rapidly degraded with a half-life of 8 to 48 hours, depending on mechanisms of introduction into water. Degradation profiles of deltamethrin in water indicated rapid conversion of deltamethrin to decamethrinic acid (BR2CA), comprising approximately 90% of the radioactivity in the aqueous phase at 168 hours. Extraction and analysis of fathead minnows (Pimephales promelas) after 96 hours revealed that tissue residues contained parent compounds and metabolites alpha-R-deltamethrin, trans-deltamethrin and Br2CA. Fish residues are directly related to aqueous concentrations, thus bioavailability under field runoff regimes were an order of magnitude lower than tissue residues under spray drift conditions. Plant tissue was found to significantly accumulate pyrethroids.     

Potential re-colonisation by cladocerans of an acidic tropical pond

To predict how re-colonisation of acidified lakes will proceed, at least two approaches are possible: (i) to compare the life history traits of candidate species and determine which one has the highest fitness, and (ii) to simulate a more realistic scenario carrying out experiments with the grouping of the candidate species, so that the intrinsic rate of natural increase of each species is integrated with its sensitivity to low pH and its ability to compete with the other candidate populations. The objective of the present study was to investigate the future re-colonisation of such acidified systems, taking as case-study a tropical pond (Lagoa das Dunas, Camaçari, BA, Brazil) and four species of cladocerans occurring in nearby water bodies (Ceriodaphniacornuta, Ceriodaphniasilvestrii, Latonopsisaustralis and Macrothrix elegans), by comparing the two above mentioned approaches. The second approach included two sets of in situ microcosms experiments, one simulating the re-colonisation by immigrating ephippia, thus using neonates of each species as colonisers, and another simulating the immigration of adults. Both these simulations followed nearly the same trends. The integration of the effects of a higher temperature, a different photoperiod and species competition determined differences in the species densities ranking between the two approaches: life history versus microcosms. The densities of C.cornuta in the microcosms matched the biphasic concentration/response hormetic model, in the simultaneous presence of two increasingly intense stressors (interspecific competition and acidity), with a low-dose stimulation and a high-dose inhibition. The present study provided, thus, a further support to the acceptance of hormesis in ecotoxicology, also at the population level in multispecies experiments.     

Mineralization of PAHs in coal-tar impacted aquifer sediments and associated microbial community structure investigated with FISH

The microbial community structure and mineralization of polycyclic aromatic hydrocarbons (PAHs) in a coal-tar contaminated aquifer were investigated spatially using fluorescence in situ hybridization (FISH) and in laboratory-scale incubations of the aquifer sediments. DAPI-detected microbial populations in the contaminated sediments were three orders of magnitude greater than nearby uncontaminated sediments, suggesting growth on coal-tar constituents in situ. Actinobacteria, beta- and gamma-Proteobacteria, and Flavobacteria dominated the in situ aerobic (>1 mg l(-1) dissolved oxygen) microbial community, whereas sulfate-reducing bacteria comprised 37% of the microbial community in the sulfidogenic region of the aquifer. Rapid mineralization of naphthalene and phenanthrene were observed in aerobic laboratory microcosms and resulted in significant enrichment of beta- and gamma-Proteobacteria potentially explaining their elevated presence in situ. Firmicutes, Flavobacteria, alpha-Proteobacteria, and Actinobacteria were also enriched in the mineralization assays, but to a lesser degree. Nitrate- and sulfate-limited mineralization of naphthalene in laboratory microcosms occurred to a small degree in aquifer sediments from locations where groundwater chemistry indicated nitrate- and sulfate-reduction, respectively. Some iron-limited mineralization of naphthalene and phenanthrene was also observed in sediments originating near groundwater measurements of elevated ferrous iron. The results of this study suggest that FISH may be a useful tool for providing a much needed link between laboratory microcosms and groundwater measurements made in situ necessary to better demonstrate the potential for natural attenuation at complex PAH contaminated sites.     

Effects of combinations of malathion and cypermethrin on survivability and time of metamorphosis of tadpoles of Indian cricket frog (Fejervarya limnocharis)

This study was undertaken to determine the effect of environmentally realistic concentrations of two commonly used pesticides viz., malathion and cypermethrin, using a fully 3 × 3 factorial experiments on the survivability and time of metamorphosis in a common rice paddy field frog (cricket frog) Fejervarya limnocharis under laboratory conditions. The results suggest that cypermethrin is more toxic than malathion and combinations of higher concentrations of cypermethrin (50 μg/L) with malathion (250 and 500 μg/L) are more deleterious to the survivability of tadpoles. With increasing cypermethrin concentration, the survivability of tadpole decreased (r = -0.986, P = 0.108). But cypermethrin alone induced early metamorphosis among the surviving tadpoles. However, there was a delay in the time required for metamorphosis induced by malathion and its combination with cypermethrin. The delay in metamorphosis may indicate the altered physiological fitness of the individual. The emergent froglets will be subjected to environmental stressors like high temperature and less humidity of post-monsoon tropical climate that could enhance negative influence triggered by pesticides.     

Unexpected effects of sublethal doses of insecticide on the peripheral olfactory response and sexual behavior in a pest insect

Pesticides have long been used as the main solution to limit agricultural pests, but their widespread use resulted in chronic or diffuse environmental pollutions, development of insect resistances, and biodiversity reduction. The effects of low residual doses of these chemical products on organisms that affect both targeted species (crop pests) but also beneficial insects became a major concern, particularly because low doses of pesticides can induce unexpected positive—also called hormetic—effects on insects, leading to surges in pest population growth at greater rate than what would have been observed without pesticide application. The present study aimed to examine the effects of sublethal doses of deltamethrin, one of the most used synthetic pyrethroids, known to present a residual activity and persistence in the environment, on the peripheral olfactory system and sexual behavior of a major pest insect, the cotton leafworm Spodoptera littoralis. We highlighted here a hormetic effect of sublethal dose of deltamethrin on the male responses to sex pheromone, without any modification of their response to host-plant odorants. We also identified several antennal actors potentially involved in this hormetic effect and in the antennal detoxification or antennal stress response of/to deltamethrin exposure.     

Review of recent ecotoxicological studies on cladocerans

Cladocerans have been widely used as the bioassay organisms in evaluating the impact of different toxic substances. Literature survey during the last 10 years revealed that cladoceran ecotoxicological research has been in an exponential phase constituting nearly 10% of publications on this group. Many studies have considered typically planktonic taxa such as Daphnia magna, D. pulex, Moina macrocopa, M. micrura and Ceriodaphania dubia. Experimental data on toxicity tests, to a lesser extent, are also available for littoral-benthic genera such as Simocephalus, Macrothrix and Alona. Most toxicity tests are limited to the derivation of median lethal concentrations of various durations but mostly at 24 or 48 h. Studies related to the evaluation of changes in the life history variables of cladocerans as a result of sublethal exposure to toxic substances are not many, but gaining importance. The common toxic substances used in the cladoceran toxicity tests appear to be heavy metals, pesticides and a few natural toxins such as cyanotoxins. We review here the effect of different toxic substances on cladocerans based on both the field and the laboratory studies from an ecotoxicology point of view. Suggestions for the future cladoceran ecotoxicology are also commented on.     

Organochlorine pesticides accumulation and degradation products in vegetation samples of a contaminated area in Galicia (NW Spain)

The content of 21 organochlorine pesticides were studied in vegetation samples of a highly contaminated area by isomers of hexachlorocyclohexane (HCH) located close to a former industrial area in Galicia (NW Spain). Five species of plants were collected at different points of the contaminated area and the different parts of the plants were separated in order to study differences in accumulation capabilities. Samples were extracted employing microwave energy followed by a clean-up step using solid phase extraction and finally determined by GC–ECD. The results obtained show that the most abundant pesticides are HCHs isomers, being the main isomers β-HCH and -HCH in all samples whereas δ-HCH and γ-HCH were at lower levels. Some other pesticides such as p,p′-DDT, p,p′-DDD and p,p′-DDE were also present in much lower amount in some of the samples. Several degradation products of HCH were also identified in some samples by GC–MS.     

Review of Recent Ecotoxicological Studies on Cladocerans

Cladocerans have been widely used as the bioassay organisms in evaluating the impact of different toxic substances. Literature survey during the last 10 years revealed that cladoceran ecotoxicological research has been in an exponential phase constituting nearly 10% of publications on this group. Many studies have considered typically planktonic taxa such as Daphnia magna, D. pulex, Moina macrocopa, M. micrura and Ceriodaphania dubia. Experimental data on toxicity tests, to a lesser extent, are also available for littoral-benthic genera such as Simocephalus, Macrothrix and Alona. Most toxicity tests are limited to the derivation of median lethal concentrations of various durations but mostly at 24 or 48 h. Studies related to the evaluation of changes in the life history variables of cladocerans as a result of sublethal exposure to toxic substances are not many, but gaining importance. The common toxic substances used in the cladoceran toxicity tests appear to be heavy metals, pesticides and a few natural toxins such as cyanotoxins. We review here the effect of different toxic substances on cladocerans based on both the field and the laboratory studies from an ecotoxicology point of view. Suggestions for the future cladoceran ecotoxicology are also commented on.     

Hexadecane and pristane degradation potential at the level of the aquifer—evidence from sediment incubations compared to in situ microcosms

Monitored natural attenuation is widely accepted as a sustainable remediation method. However, methods providing proof of proceeding natural attenuation within the water-unsaturated (vadose) zone are still relying on proxies such as measurements of reactive and non-reactive gases, or sediment sampling and subsequent mineralisation assays, under artificial conditions in the laboratory. In particular, at field sites contaminated with hydrophobic compounds, e.g. crude oil spills, an in situ evaluation of natural attenuation is needed, because in situ methods are assumed to provide less bias than investigations applying either proxies for biodegradation or off-site microcosm experiments. In order to compare the current toolbox of methods with the recently developed in situ microcosms, incubations with direct push-sampled sediments from the vadose and the aquifer zones of a site contaminated with crude oil were carried out in conventional microcosms and in situ microcosms. The results demonstrate the applicability of the in situ microcosm approach also outside water-saturated aquifer conditions in the vadose zone. The sediment incubation experiments demonstrated turnover rates in a similar range (vadose, 4.7 mg/kg*day; aquifer, 6.4 mg_hexadecane/kg_soil/day) of hexadecane degradation in the vadose zone and the aquifer, although mediated by slightly different microbial communities according to the analysis of fatty acid patterns and amounts. Additional experiments had the task of evaluating the degradation potential for the branched-chain alkane pristane (2,6,10,14-tetramethylpentadecane). Although this compound is regarded to be hardly degradable in comparison to n-alkanes and is thus frequently used as a reference parameter for indexing the extent of biodegradation of crude oils, it could be shown to be degraded by means of the incubation experiments. Thus, the site had a high inherent potential for natural attenuation of crude oils both in the vadose zone and the aquifer.     

Pesticide fate in tropical wetlands of Brazil: an aquatic microcosm study under semi-field conditions

A contamination of off-site aquatic environments with pesticides has been observed in the tropics, yet only sparse information exists about pesticide fate in such ecosystems. The objective of our semi-field study was to elucidate the fate of alachlor, atrazine, chlorpyrifos, endosulfan, metolachlor, profenofos, simazine, and trifluralin in the aqueous environment of the Pantanal wetland (MT, Brazil). To this aim, water and water/sediment microcosms of two sizes (0.78 and 202 l) were installed in the outskirts of this freshwater lagoon environment and pesticide dissipation was monitored for up to 50 d after application. The physical-chemical water conditions that developed in the microcosms were reproducible among field replicates for both system sizes. Pesticide dissipation was substantially enhanced for most pesticides in small microcosms relative to the large ones (reduced DT(50) by a factor of up to 5.3). The presence of sediment in microcosms led to increased persistence of chlorpyrifos, endosulfan, and trifluralin in the test systems, while for polar pesticides (alachlor, atrazine, metolachlor, profenofos, and simazine) a lesser persistence was observed. Atrazine, simazine, metolachlor, and alachlor were identified as the most persistent pesticides in large water microcosms (DT(50) > or = 47 d); in large water/sediment systems endosulfan beta, atrazine, metolachlor, and simazine showed the slowest dissipation (DT(50) > or = 44 d). A medium-term accumulation in the sediment of tropical ecosystems can be expected for chlorpyrifos and endosulfan isomers (11-35% of applied amount still extractable at 50 d after application). We conclude that the persistence of the studied pesticides in aquatic ecosystems of the tropics is not substantially lower than during summer in temperate regions.     

Partial compensation of the sublethal effect of deltamethrin on the sex pheromonal communication of Trichogramma brassicae

Pyrethroid insecticides are widely used and lead to a sizable environmental pollution that could interfere with the population biology of insects. Trichogramma is a beneficial insect used in biological control and which natural populations contribute to the control of Lepidopterus pests. In this work, we determined the effect of a sublethal dose of deltamethrin on the sex pheromonal communication of Trichogramma. The dose used (LD 0.1) induces no detectable mortality (the theoretical mortality is only one insect over 1000) and can be a good representation of contamination by this insecticide from environmental pollution. The insecticide was shown to have opposite effects on the sex pheromonal communication of Trichogramma, depending on which sex was exposed (Delpuech, J.M., Legallet, B., Terrier, O., Fouillet, P., 1999. Chemosphere 38, 729–739). We show that, when both sexes are simultaneously exposed to the insecticide, this effect is only partially neutralized. The mean response of treated males responding to the sex pheromone from treated females is not significantly different from that of controls, but the kinetics of their response is not the same. When both sexes are treated, the response of males to the sex pheromone is lower at the beginning but their response does not decrease during time contrary to controls and becomes finally higher than that of controls. Therefore, the sublethal effect of deltamethrin in the field can be either advantageous or disadvantageous depending on the difficulty in finding females and their scarcity.     

Evaluation of TCDD biodegradability under different redox conditions

Polychlorinated dibenzo-p-dioxins have been generated as unwanted by-products in many industrial processes. Although their widespread distribution in different environmental compartments has been recognized, little is known about their fate in the ultimate environment sinks. The highly stable dioxin isomer 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been called the most toxic compound known to man. In this laboratory microcosm study, TCDD bioavailability was evaluated under five reduction/oxidation (redox) conditions including aerobic biodegradation, aerobic cometabolism, methanogenesis, iron reduction, and reductive dechlorination. Activated sludge and aquifer sediments from a TCDD and a pentachlorophenol (PCP) contaminated site were used as the inocula. Acetate, sludge cake, and cane molasses were used as the primary substrates (carbon sources) in cometabolism and reductive dechlorination microcosms. After a 90-day incubation period, microcosms constructed under reductive dechlorination conditions were the only treatment showing promising remediation results. The highest TCDD degradation rate [up to 86% of TCDD removal (with an initial concentration of 96 microg/kg of soil)] was observed in the microcosms with anaerobic activated sludge as the microbial inocula and sludge cakes as the primary substrates. Except for reductive dechlorination microcosms, no significant TCDD removal was observed in the microcosms prepared under other conditions. Thus, application of an effective primary substrate to enhance the reductive dechlorination process is a feasible method for TCDD bioremediation. Bioremediation expense can be significantly reduced by the supplement of some less expensive alternative substrates (e.g., sludge cakes, cane molasses). Results would be useful in designing a scale-up in situ or on-site bioremediation system such as bioslurry reactor for field application.     

Microcosm investigations of stormwater pond sediment toxicity to embryonic and larval amphibians: variation in sensitivity among species

Stormwater ponds have become common features of modern development and often represent significant amounts of open space in urbanized areas. Although stormwater ponds may provide habitat for wildlife, factors responsible for producing variation in wildlife use of ponds have received limited attention. To investigate the role of variation in species tolerances of pollutants in structuring pond-breeding amphibian assemblages, we exposed species tolerant (Bufo americanus) and not tolerant (Rana sylvatica) of urbanization to pond sediments in laboratory microcosms. Pond microcosms had elevated sediment metal levels and chloride water concentrations. Among R. sylvatica embryos, exposure to pond sediments resulted in 100% mortality. In contrast, B. americanus embryos and larvae experienced only sublethal effects (i.e., reduced size at metamorphosis) due to pond sediment exposure. Our results suggest variation in pollutant tolerance among early developmental stages of amphibians may act in concert with terrestrial habitat availability to structure amphibian assemblages associated with stormwater ponds.     

Comparison of an assay for Dehalococcoides DNA and a microcosm study in predicting reductive dechlorination of chlorinated ethenes in the field

The study aims to compare the detection of 16S rRNA gene of Dehalococcoides species and the microcosm study for biotransformation in predicting reductive dechlorination of chlorinated ethenes in ground water at hazardous waste sites. A total of 72 ground water samples were collected from 12 PCE or TCE contaminated sites in the United States. The samples were analyzed and used to construct microcosms in the laboratory. The results showed that the presence of Dehalococcoides DNA was well associated with dechlorination to ethene in the field. Nearly half of the wells where Dehalococcoides DNA was detected had ethene as a dechlorination end product. In comparison, for ground water samples of 16 wells where ethene was detected, ethene was produced in 11 of the corresponding microcosms. For most microcosms, during two years of incubation, dechlorination was less extensive than that observed in the field.