A pond mesocosm study to determine direct and indirect effects of lindane on a natural zooplankton community

We assessed the threshold concentrations for toxic effects by lindane on a zooplankton community using mesocosms in a natural pond. Enclosures (1000 litres) were treated to give mean concentrations of 0, 2, 6, 12, 18, 24, and 50 microg litre(-1) over a test period of 39 days. Larvae of the phantom midge Chaoborus flavicans showed a high susceptibility to lindane and were virtually eliminated from enclosures with > or =12 microg litre(-1) 4 days after application. Populations of cyclopoid nauplii (Eucyclops serrulatus) were reduced at a lindane concentration of 2 microg litre(-1) and showed high mortality after 11 days exposure to > or =6 g litre(-1). No lindane-induced effects could be seen in the cladoceran species Daphnia longispina and Chydorus sphaericus. Rotifer species were probably not directly affected by lindane; however loss of the principal predator Chaoborus was accompanied by an increase in the numbers of Asplanchna priodonta. A simultaneous decrease in Keratella quadrata possibly reflected predation since its decline showed no dose-response relationship with lindane and it is among the major prey of Asplanchna. Such complex secondary effects, occurring through trophic interactions between a predator and two co-occurring prey, would not have been predicted from single-species toxicity tests. These, and effects on cyclopoid populations that occurred predominantly through the mortality of nauplii, demonstrate the value of toxicological studies on natural communities. We conclude that a combination of laboratory and field investigations, coupled with mono-species and community assessments, provide the best route for understanding the effects of toxicants such as lindane for regulatory purposes.     

Relative sensitivity of Daphnia magna and Brachionus calyciflorus to five pesticides.

Comparative toxicity of several pesticides, lindane, endosulfan, pentachlorophenol (PCP),3,4-dichloroaniline (DCA) and copper sulphate has been tested to determine their lethality in two species of freshwater invertebrates, Daphnia magna and Brachionus calyciflorus. D. magna was more sensitive than B. calyciflorus to all the toxicants tested except to copper sulphate. DCA was the most toxic compound to D. magna and was followed in order of decreasing toxicity by copper sulphate, PCP, endosulfan and lindane. Copper sulphate was the most toxic chemical tested to the rotifer B. calyciflorus and was followed by PCP, endosulfan, lindane and DCA. The 24 hr LC50 values (mg/L) for D. magna and B. calyciflorus, respectively, were: lindane, 1.64 and 22.5; endosulfan, 0.62 and 5.15; DCA, 0.20 and 61.5; PCP, 0.39 and 2.16; copper sulphate, 0.38 and 0.076.     

Toxicity of nanosized and bulk ZnO, CuO and TiO2 to bacteria Vibrio fischeri and crustaceans Daphnia magna and Thamnocephalus platyurus

As the production of nanoparticles of ZnO, TiO2 and CuO is increasing, their (eco)toxicity to bacteria Vibrio fischeri and crustaceans Daphnia magna and Thamnocephalus platyurus was studied with a special emphasis on product formulations (nano or bulk oxides) and solubilization of particles. Our innovative approach based on the combination of traditional ecotoxicology methods and metal-specific recombinant biosensors allowed to clearly differentiate the toxic effects of metal oxides per se and solubilized metal ions. Suspensions of nano and bulk TiO2 were not toxic even at 20 g l(-1). All Zn formulations were very toxic: L(E)C50 (mg l(-1)) for bulk ZnO, nanoZnO and ZnSO4.7H2O: 1.8, 1.9, 1.1 (V. fischeri); 8.8, 3.2, 6.1 (D. magna) and 0.24, 0.18, 0.98 (T. platyurus), respectively. The toxicity was due to solubilized Zn ions as proved with recombinant Zn-sensor bacteria. Differently from Zn compounds, Cu compounds had different toxicities: L(E)C50 (mg l(-1)) for bulk CuO, nano CuO and CuSO4: 3811, 79, 1.6 (V. fischeri), 165, 3.2, 0,17 (D. magna) and 95, 2.1, 0.11 (T. platyurus), respectively. Cu-sensor bacteria showed that toxicity to V. fischeri and T. platyurus was largely explained by soluble Cu ions. However, for Daphnia magna, nano and bulk CuO proved less bioavailable than for bacterial Cu-sensor. This is the first evaluation of ZnO, CuO and TiO2 toxicity to V. fischeri and T. platyurus. For nano ZnO and nano CuO this is also a first study for D. magna.     

Is Daphnia magna an ecologically representative zooplankton species in toxicity tests?

Daphnia magna is commonly used in aquatic toxicity testing because of many characters that make it easy and economical to culture in the laboratory: it is relatively small, has short life cycle, high fecundity, and parthenogenetic reproduction. On the other hand, D. magna differs from other freshwater zooplankters in size, habitat, life-history, and ability to withstand fish predation. D. magna is a relatively large zooplankton species which makes it so vulnerable to fish predation that it is excluded from fish-inhabiting lakes. It occurs mainly in ephemeral habitats like small ponds and rockpools where vertebrate predators are rare. As a result, D. magna is seldom an indigenous species in lakes which receive pollutants, although representativeness is one important criterion for the standardised toxicity test species. Small ponds are unpredictable habitats with large temporal and spatial variability in abiotic factors. Adaptation to this natural abiotic stress may increase pollution tolerance. The life-history of D. magna also differs from that of lake-inhabiting cladocerans. Large daphnids produce many small neonates, whereas the opposite is true for small cladocerans. The large neonate size allows an earlier maturation of small cladocerans compared to daphnids. In a few comparative studies D. magna tended to be less sensitive to toxic substances than other cladocerans, and this may be due in part to life-history and size differences.     

Accumulation and elimination of aqueous and dietary silver in Daphnia magna

The dissolved uptake, dietary assimilation, and efflux of Ag in a freshwater cladoceran, Daphnia magna, were measured under different laboratory conditions. The dissolved uptake rate of Ag was proportional to the ambient Ag concentration, but the accumulation was highly variable due to the sorption of Ag onto the daphnid bodies. The ambient Na(+) but not the ambient K(+) concentration significantly decreased the dissolved uptake of Ag, suggesting a competitive uptake of Ag(+) with Na(+). The dietary assimilation efficiencies (AEs) of Ag are dependent on the concentration of the algal food available to D. magna. The AE was as low as 2% when the food concentration reached the saturation levels. In contrast, the Ag concentrations in the algae did not significantly affect the Ag AE in D. magna. The efflux rate constant of diet-incorporated Ag was twice that through dissolved uptake. The elimination of Ag was further separated into different compartments (excretion, egestion, molting, and reproduction) in the juveniles and adults after accumulation from dissolved and dietary sources. Regeneration into the dissolved phase was the predominant pathway by which the incorporated Ag was lost from D. magna, regardless of the exposure pathway. In contrast to the essential metals or Hg, there was minimal maternal transfer of Ag from the mothers to the offspring. By employing the biokinetic model, we further showed that water is a dominant pathway for Ag accumulation in D. magna. Trophic transfer is less significant primarily because of the low Ag AE when the food concentration reached the saturation levels.     

Adaptation to environmental stress in Daphnia magna simultaneously exposed to a xenobiotic

In standardized ecotoxicological testing chemicals are investigated under optimal conditions for the test organisms despite the fact that environmental factors such as predation pressure and food availability are important parameters regulating natural populations. Food limitation and predator presence can induce shifts in life-history traits in various Daphnia species, especially trade-offs in reproductive biomass allocation. These adaptive responses are thought to ensure survival of the population in a highly variable environment. A xenobiotic dispersant (used in textile dyeing processes) also shifted the biomass allocation of Daphnia magna. To assess whether the dispersant could hinder D. magna adaptation to varying environmental conditions, we conducted experiments with food level and presence of Chaoborus larvae as environmental factors and simultaneous exposure to the dispersant. At low food level and in presence of the predator, D. magna produced fewer but larger sized neonates, regardless of dispersant exposure. The dispersant shifted biomass allocation towards more but smaller sized offspring in all experiments. However, the adaptive response to the environmental factors and the dispersant effect cancelled each other out in that they induced independently from each other opposite shifts in biomass allocation. In summary, the dispersant exposure resulted not in an inhibition of the adaptive response but in a reduction of the value of the response. Our study with this model substance demonstrates that xenobiotics can affect the adaptation of organisms to environmental stress which can result in effects likely to be overlooked in standardized testing.     

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.     

Toxicity of fluoranthene and its biodegradation metabolites to aquatic organisms

The toxicity of nine stable products of the biodegradation of fluoranthene with the pure bacterial strain Pasteurella sp. IFA was studied. For their quantification, an improved analytical procedure with two-step liquid-liquid extraction, derivatisation and gas chromatographic-mass spectrometric detection was used. Growth inhibition and immobility tests for fluoranthene and its metabolites were carried out using algae (Scenedesmus subspicatus), bacteria (Pseudomonas putida) and crustaceans (Daphnia magna and Thamnocephalus platyurus). Tests using the alga S. subspicatus revealed that with the exception of 9-hydroxyfluorene, which was only four times less toxic than fluoranthene, all the other metabolites were 37 to approximately 3000 times less toxic than the parent material. P. putida cells were resistant to fluoranthene and its primary metabolites, but were inhibited by low molecular weight intermediates, especially benzoic acid. Fluoranthene was not toxic to T. Platyurus, but was toxic to D. magna. Its primary metabolites (including 9-fluorenone and 9-hydroxyfluorene) were toxic to D. magna, and a low molecular weight metabolite (2-carboxybenzaldehyde) was highly toxic to T. platyurus.     

Acute toxicity of leachates of tire wear material to Daphnia magna--variability and toxic components

Large amounts of tire rubber are deposited along the roads due to tread wear. Several compounds may leach from the rubber and cause toxicity to aquatic organisms. To investigate the toxic effects of tire wear material from different tires, rubber was abraded from the treads of twenty-five tires. Leachates were prepared by allowing the rubber to equilibrate with dilution water at 44 degrees C for 72 h. Then the rubber was filtered from the leachates, and test organisms (Daphnia magna) were added. Forty-eight hour EC50s ranged from 0.5 to >10.0 g l(-1). The toxicity identification evaluation (TIE) indicated that non-polar organic compounds caused most of the toxicity. UV exposure of the filtered tire leachates caused no significant increase in toxicity. However, when tested as unfiltered leachates (the rubber was not filtered from the leachates before addition of D. magna) photo-enhanced toxicity was considerable for some tires, which means that test procedures are important when testing tire leachates for aquatic (photo) toxicity. The acute toxicity of tire wear for Daphnia magna was found to be <40 times a predicted environmental concentration based on reports on the concentration of a tire component found in environmental samples, which emphasizes the need for a more extensive risk assessment of tire wear for the environment.     

Effects of carbaryl on the spring zooplankton communities in ponds

A carbamate insecticide, carbaryl, was applied in spring to concrete ponds to study its effects on zooplankton communities. The population density of Cladocera (Daphnia spp.) was nearly constant before application of the chemical. Carbaryl at 1 ppm killed all zooplankton species, including Chaoborus larvae. After treatment, cladocerans never recovered to their previous level. The relatively rapid recovery of a predator, Chaoborus, seemed to interfere with recovery of the cladoceran populations. The lower water temperature occurring in spring was thought to favour the former because of its influence on growth rates.     

Acute effects of diclofenac and DMSO to Daphnia magna: immobilisation and hsp70-induction

To determine the toxicity of the anti-rheumatic drug diclofenac to Daphnia magna, acute toxicity tests according to the OECD guideline 202 were combined with biochemical investigations of the hsp70 level as a biomarker for proteotoxicity. Particular attention was paid to the impact of the solvent DMSO as a confounding factor to diclofenac toxicity by means of testing different variations of producing stock solutions. In the acute immobilisation tests, diclofenac was most toxic as a singular test substance, with indication of a slight antagonistic interaction between the two substances. The highest EC(50) values were obtained in those approaches using diclofenac pre-dissolved in DMSO. Thus, the observed antagonism seems to be intensified by pre-dissolution. Hsp70 levels of 12- to 19-days-old D. magna were determined after 48h exposure using a highly reproducible immunological protocol. Hsp70 induction occurred at a LOEC of 30mgl(-1) diclofenac plus 0.6mll(-1) DMSO, and at a LOEC of 40mgl(-1) for diclofenac alone. In summary, DMSO showed only slight confounding effects on diclofenac action in the applied range of concentrations.     

Effects of pharmaceuticals on Daphnia survival, growth, and reproduction

Pharmaceuticals have been globally detected in surface waters, and the ecological impacts of these biologically-active, ubiquitous chemicals are largely unknown. To evaluate the aquatic toxicity of individual pharmaceuticals and mixtures, we performed single species laboratory toxicity tests with Daphnia magna, a common freshwater zooplankton. We conducted acute (6-day) and chronic (30-day) exposure pharmaceutical bioassays and evaluated survivorship and morphology of adults and neonates, adult length, resting egg production, brood size (fecundity), and the proportion of male broods produced (sex ratio). In general, exposure to a single pharmaceutical in the 1-100 microg/l range yielded no apparent effects on the normal life processes of Daphnia. However, chronic fluoxetine exposure (36 microg/l) significantly increased Daphnia fecundity, and acute clofibric acid exposure (10 microg/l) significantly increased sex ratio. A mixture of fluoxetine (36 microg/l) and clofibric acid (100 microg/l) caused significant mortality; the same fluoxetine concentration mixed with 10 microg/l clofibric acid resulted in significant deformities, including malformed carapaces and swimming setae. Mixtures of three to five antibiotics (total antibiotic concentration 30-500 microg/l) elicited changes in Daphnia sex ratio. We conclude: (1) individual and mixtures of pharmaceuticals affect normal development and reproduction of Daphnia magna, (2) aquatic toxicity of pharmaceutical mixtures can be unpredictable and complex compared to individual pharmaceutical effects, and (3) timing and duration of pharmaceutical exposure influence aquatic toxicity.     

Effects of the parasiticide ivermectin on the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata

Although widely used for the treatment of endo- and ectoparasites in livestock and pets, very few data on chronic effects on aquatic organisms are available for the parasiticide ivermectin. In the present study, toxicity of ivermectin to two freshwater organisms, the cladoceran Daphnia magna and the green alga Pseudokirchneriella subcapitata was investigated. For D. magna, a mean LC(50) 48 h of 5.7 ngl(-1) was derived from 10 acute tests. Chronic toxicity of ivermectin to D. magna was extremely high: with 0.001 and 0.0003 ngl(-1), respectively, nominal LOEC and NOEC based on growth and reproduction were far below the analytical limit of detection for this compound. P. subcapitata was considerably less sensitive to ivermectin than D. magna. For both growth rate and yield, EC(50) was >4,000 microgl(-1), LOEC was 1,250 microgl(-1) and NOEC 391microgl(-1). In view of the high toxicity to D. magna, the use of ivermectin might pose a risk to local aquatic ecosystems. Further studies should be carried out to investigate the effects of ivermectin and its degradation products on pelagic and benthic freshwater invertebrates.     

Comparative toxicity of imidacloprid, of its commercial liquid formulation and of diazinon to a non-target arthropod, the microcrustacean Daphnia magna

Imidacloprid (IMI) is at the moment the insecticide with the world's fastest growing sales and is considered possible replacement for the widely used organophosphorus pesticide, diazinon, which is subject to phased revocation in many countries. In this study, biochemical, reproductive and survival parameters of the water flea (Daphnia magna) after chronic exposure to IMI, its commercial liquid formulation Confidor SL 200 and diazinon are presented and compared. According to the lowest observed effect concentrations, diazinon is more toxic to the reproduction of D. magna than IMI and Confidor SL 200, which exert similar toxicity. The same was observed for the survival, except that Confidor SL 200 is more toxic than IMI. In polluted aquatic environments, the actual levels of diazinon are potentially chronically hazardous to the reproduction of D. magna (risk quotient >1). According to very few measured environmental levels of IMI, the latter is not expected to be chronically hazardous, unless it is accidentally spilled in a small pond. In such case, the predicted concentrations of IMI would present a potential chronic risk to D. magna, and a potential acute risk to other aquatic invertebrates. In the future, higher environmental levels of IMI are expected due to its increasing use and physico-chemical properties. The literature survey summarized in this work suggests that further ecotoxicological studies with a broader spectrum of aquatic organisms are needed before IMI is classified as safer than currently applied pesticides.     

Determination photostability of selected agrochemicals in water and soil.

The photolysis of selected pesticides in aqueous solutions has been investigated. The photolysis produced different intermediate substances, which were also found to be soil and microbial degradation products. The phototransformation in the presence of TiO2 and humic substances leads to a disappearance of these compounds. The reaction rate is dependent on the semiconductor oxide and concentration. Photoproducts were isolated and characterized by different spectroscopic methods. Results from this study indicate that degradation products of isoproturon are more toxic on Daphnia magna than on the parent compound.     

Assessment of toxicant activity in sediments by the ECHA biocide monitor

The ECHA Biocide Monitor, a dipstick procedure originally designed to test the effectiveness of in situ biocides, was evaluated as a possible environmental toxicity assessment procedure. The dipstick procedure was applied to 94 sediment extracts which were also tested by three well documented toxicant screening procedures; Microtox, Daphnia magna and Spirillum volutans. In these samples the Daphnia magna test proved to be the most sensitive indication of toxicant activity (63 of 94) with the ECHA Biocide Monitor easily being the second most sensitive procedure with 27 of 94 sediments indicating the presence of toxic substances. Results of the investigation are detailed.     

Biodegradation in laboratory activated sludge plants and aquatic toxicity of herbicides

The biodegradation and the aquatic toxicity of four herbicides (isoproturon, terbuthylazine, mecoprop, metamitron) were investigated. Laboratory activated sludge plants were used for biodegradation experiments. The biodegradation of mecoprop reached nearly 100%, the other herbicides were not eliminated by biodegradation. The acute Daphnia magna 24-h assay, the algal 72-h inhibition test, and the recently developed lemna growth inhibition 7-d test were applied to evaluate the biological effects of herbicides as original substances. EC 50 and EC 10 values were determined. Algal and lemna test show that isoproturon and terbuthylazine are both much more toxic than mecoprop and metamitron. Daphnids are generally less sensitive against herbicides than plants. Biodegradation and toxicity test were coupled for mecoprop to assess biological long-term effects of possible biodegradation products of this herbicide. The effluents of the laboratory activated sludge units were used in toxicity tests (Daphnia magna 21-d reproduction test, lemna growth inhibition 7-d test). No inhibiting effect on the tested organisms was observed.     

Acute and chronic toxicity of veterinary antibiotics to Daphnia magna

The acute and chronic toxicity of nine antibiotics used both therapeutically and as growth promoters in intensive farming was investigated on the freshwater crustacean Daphnia magna. The effect of the antibiotics metronidazole (M), olaquindox (OL), oxolinic acid (OA), oxytetracycline (OTC), streptomycin (ST), sulfadiazine (SU), tetracycline (TC), tiamulin (TI) and tylosin (TY) was tested in accordance to the ISO (1989) and OECD (1996) standard procedures. The acute toxicities (48-h EC50 value, mg/l) in decreasing order were OA (4.6), TI (40), SU (221), ST (487), TY (680) and OTC (approximately 1000). NOECs were 340 mg/l for TC and 1000 mg/l for M and OL. Toxic effect on reproduction occurred generally at concentrations, which were one order of magnitude below the acute toxic levels. The chronic toxicity (EC50 values, mg/l) in the D. magna reproduction test in decreasing order were TI (5.4), SU (13.7), TC (44.8) and OTC (46.2). The NOECs (mg/l) obtained in the reproduction test with OA, ST, TY and M were 0.38 for OA, 32 for ST, 45 for TY and 250 for M. The observed toxicity of OA to D. magna indicates that this substance, which is a commonly used feed additive in fish farms, has a potential to cause adverse effects on the aquatic environment.     

Relative sensitivity of one freshwater and two marine acute toxicity tests as determined by testing 30 offshore E & P chemicals

Acute toxicity of 30 offshore E & P (Exploration and Production) chemicals was measured using the three standard test organisms Daphnia magna (freshwater cladoceran), Acartia tonsa (marine copepod) and Skeletonema costatum (marine diatom alga). Test chemicals included 20 water-soluble and 10 (partially) non-soluble products. For 22 out of the 30 chemicals, the difference in sensitivity between the three tests varied within one order of magnitude. A very good correlation was found between the two marine tests (r = 0.96, P < 0.01, n = 30), and a correlation coefficient of r = 0.78 (P < 0.01, n = 30) was found between D. magna and both A. tonsa and S. costatum, individually. When the comparison of D. magna and A. tonsa sensitivity was based only on the water-soluble chemicals, a significantly higher correlation was obtained (r = 0.84, n = 20), indicating that the sample preparation method used for the (partially) non-soluble chemicals (the water accommodated fraction (WAF) method) induces additional variation between tests performed with different test media. (Partially) non-soluble chemicals are characterised by phase separation or precipitation at the concentrations used for testing. In a WAF-based test, each test concentration/exposure level is prepared separately, and following mixing and separation, only the water phase is used for testing. Toxicity is related to the amount of substance originally added to the mixing vessels. For 25 of the 30 chemicals, D. magna was found to be less sensitive than the marine copepod by a factor >2. The generally higher sensitivity of the marine toxicity tests compared to the Daphnia test emphasise the importance of using marine data for environmental hazard classification as well as for environmental risk assessment purposes.     

Multi-generational effects of four selected environmental oestrogens on Daphnia magna

The objective of this study was to determine whether vertebrate-type oestrogens have ecotoxicological effects on a crustacean species. The effects of 17beta-oestradiol (E2), diethylstilbestrol (DES), bisphenol A (BPA) and 4-nonylphenol (4-NP) on the freshwater invertebrate Daphnia magna were assessed over first and second generations. The acute EC(50) 48 h, based on immobilisation, for E2, DES, BPA and 4-NP were 2.87 mg/l, 1.55 mg/l, 7.75 mg/l and 0.13 mg/l, respectively. The impact of the test chemicals on moulting frequency was also assessed. The EC(50) 48 h, based on the inhibition of moult number for E2, DES and 4-NP were 2.04 mg/l, 1.87 mg/l and 0.14 mg/l, respectively. BPA was not observed to impact the moulting frequency of D. magna at concentrations tested. In a series of separate studies, the effects of the four selected test compounds on the survival, moulting frequency and reproduction of first and second generational D. magna were assessed over a period of 21 d. Exposure of D. magna to 4-NP decreased the number of offspring produced in both first and second generation testing. DES proved to have no significant (p0.05) inhibition of fecundity in first generation but when second generation daphnids were exposed to DES, a significant (p0.05) reduction in the number of offspring was recorded. When D. magna were exposed to E2 or BPA, no statistically significant (p0.05) inhibition in the number of moults or offspring produced was observed.