Measurement of movement patterns of Caenorhabditis elegans (Nematoda) with the Multispecies Freshwater Biomonitor® (MFB)—a potential new method to study a behavioral toxicity parameter of nematodes in sediments

The nematode Caenorhabditis elegans receives increasing attention in sediment ecotoxicology and new toxicity tests with sensitive test parameters are under development. In this study, the motility of C. elegans could be measured for the first time online in sediment, using the Multispecies Freshwater Biomonitor. Whereas single nematodes could not be recorded, groups of 10 nematodes gave typical locomotive signals in different media (water, agar, sediment) with comparable precision and accuracy. The results of this study encourage to develop a new rapid online whole-sediment toxicity test with behaviour as sensitive test parameter.     

Chronic toxicity of sediment-associated linear alkylbenzene sulphonates (LAS) to freshwater benthic organisms

The toxicity of linear alkylbenzene sulphonates (LAS), to freshwater benthic organisms was assessed during exposure to spiked sediment. Lethal and sub-lethal end-points were monitored for two organisms (oligochaete Lumbriculus variegatus and nematode Caenorhabditis elegans). Results demonstrated relatively low toxicity (LOECs>100 mg/kg dry weight). No observed effect concentrations (NOECs) of 81 mg/kg dw (Lumbriculus) and 100 mg/kg dw (Caenorhabditis) were determined. For the oligochaete, no specific endpoint was particularly sensitive to LAS. For the nematode, egg production was the most sensitive endpoint. Significant degradation was measured over the 28-day duration of the Lumbriculus study, equating to a half-life of 20 days in sediment.     

Experimental effects of tributyltin (TBT) contaminated sediment on a range of meiobenthic communities

Microcosm experiments have been carried out with whole natural meiobenthic communities to look at the effects of TBT sediment contamination on the community structure of the dominant nematode component of the meiobenthos. TBT has a high affinity for aquatic sediments, yet this is the first study of the effects of this contaminant in sediment on natural benthic communities. Three communities were studied from contrasting locations in south-west England: the intertidal of the Lynher estuary (muddy sediment) and the Exe estuary (sandy sediment) and the subtidal (50m depth) at Rame Head off Plymouth (muddy sand). Fresh sediment with natural meiobenthic communities was incubated for 2 months with TBT-contaminated sediment (three dose levels) in bottles. Nematodes were identified and enumerated and subjected to multivariate data analysis. The sandy Exe estuary fauna was significantly affected by TBT-contaminated sediment at all three doses (0.3, 0.6 and 0.9 microg g(-1) dry wt (as Sn) sediment), whereas the offshore fauna from Rame Head was significantly affected only at the highest dose. The muddy Lynher estuary meiofauna was affected (somewhat peculiarly) at the medium dose level only. Meiobenthic nematodes may not be as sensitive to TBT-contaminated sediment as other infaunal benthos but exhibited responses to levels of contamination still persisting in some UK estuaries and harbours. Comparing the effects of TBT with those of copper and zinc in the same laboratory experiments, our observations suggest that the relative impact of TBT on meiobenthic community structure is not as great as these contaminants in marine sediments. Although there are very few observations of TBT toxicity in sediment, it appears that TBT is toxic at much lower concentrations in seawater (ppb) than it is in sediment (ppm).     

Mutatox test: a new test for monitoring environmental genotoxic agents

In this study, Yamaska River water and Milli-Q water and organically extracted sediment extracts were used to evaluate the sensitivity of a new genotoxicity screening test, the Mutatox test. Also in this study, the samples were tested for acute and chronic toxicity using the following screening test procedures: Microtox, Daphnia magna, Ceriodaphnia reticulata and ATP-TOX Systems. The Mutatox test is based on the use of a dark mutant strain of Photobacterium phosphoreum and is sensitive to chemicals which are (1) DNA damaging agents (2) DNA intercalating agents, (3) DNA synthesis inhibitors and (4) direct mutagens. In this study, the Mutatox test was found to be a simple-to-perform sensitive procedure which added greater scope to the battery of tests approach. Preliminary indications are that this procedure may prove to be one of the more responsive and valuable tests in the 'battery of tests' approach to environmental screening.     

Ecotoxicological evaluation of industrial port of Venice (Italy) sediment samples after a decontamination treatment

This work assesses the ecotoxicological effects of polluted sediment after a decontamination treatment process using a new sediment washing technique. Sediment samples were collected from four sites in Marghera Port industrial channels (Venice, Italy). Ecotoxicological evaluations were performed with Vibrio fischeri and Crassostrea gigas bioassays. Whole sediment and elutriate were deemed as the most suitable environmental matrices for this study. Toxicity scores developed in the Lagoon of Venice for V. fischeri on whole sediment and for C. gigas on elutriate were considered for the final ranking of samples. Ecotoxicological results showed that the treated sediment samples presented both acute and sub-chronic toxicities, which were mainly attributed to the presence of some remaining chemicals such as metals and polyaromatic hydrocarbons. The acute toxicity ranged from low to medium, while the sub-chronic one from absent to very high, suggesting that treated sediments could not be reused in direct contact with seawater.     

Comparison of heavy metal toxicity in life stages (spermiotoxicity, egg toxicity, embryotoxicity and larval toxicity) of Hydroides elegans

A toxicity test was developed to examine the effects of heavy metal contaminants on the early life stages of the marine polychaete. We have studied the effects of metals on fertilization and early development of marine polychaete Hydroides elegans. These heavy metals have often been found in polluted ground and water near industrial discharges, and have therefore been detected from time to time in the food chain. They have been reported to alter various reproduction functions in various animals including marine populations. The toxic effect of mercury, cadmium, lead, nickel and zinc on sperm viability, fertilization, embryogenesis and larvae of H. elegans was examined. We observed that the rate of fertilization decreased when the sperm was incubated with heavy metals. Treatment of eggs with each metal did not prevent fertilization, but delayed or blocked the first mitotic divisions, and altered early embryonic development. All these effects were observed at relatively high concentrations. However, bio-accumulation in sediments and aquatic organisms have been reported. Polychaete eggs may then be in contact with very high concentrations of these heavy metals in areas where these metals are not handled or stocked properly, and then develop into abnormal embryos. In addition to bivalves and sea-urchins, polychaete embryos can provide biological criteria for seawater quality standards taking into account the sensitivity of the invertebrates and their contribution in detection of harmful chemicals with no marked effect on the species. Our results indicate that the early development of H. elegans is highly sensitive to heavy metals and this polychaete can be routinely employed as a test organism for ecotoxicity bioassays in tropical and subtropical regions.     

Toxicity of the mosquito control pesticide Scourge to adult and larval grass shrimp (Palaemonetes pugio)

This study investigated the toxicity of various concentrations of technical resmethrin and Scourge on adult and larval Palaemonetes pugio, a common grass shrimp species. Two types of tests were conducted for each of the resmethrin formulations using adult and larval grass shrimp life stages, a 96-h static renewal aqueous test without sediment, and a 24-h static nonrenewal aqueous test with sediment. For resmethrin, the 96-h aqueous LC50 value for adult shrimp was 0.53 microg/L (95% confidence interval (CI): 0.46-0.60 microg/L), and for larval shrimp was 0.35 microg/L (95% CI: 0.28-0.42 microg/L). In the presence of sediment, technical resmethrin produced a 24-h LC50 value for adult shrimp of 5.44 microg/L (95% CI: 4.52-6.55 microg/L), and for larval shrimp of 2.15 microg/L (95% CI: 1.35-3.43 microg/L). For Scourge, the 96-h aqueous LC50 for adult shrimp was 2.08 microg/L (95% CI: 1.70-2.54 microg/L), and for larval shrimp was 0.36 microg/L (95% CI: 0.24-0.55 microg/L). The 24-h sediment test yielded an LC50 value of 16.12 microg/L (95% CI: 14.79-17.57 microg/L) for adult shrimp, and 14.16 microg/L (95% CI: 12.21-16.43 microg/L) for larvae. Adjusted LC50 values to reflect the 18% resmethrin concentration in Scourge are 0.37 microg/L (adult), 0.07 microg/L (larvae) for the 96-h aqueous test, and 2.90 microg/L (adult), 2.6 microg/L (larvae) for the 24-h sediment test. Larval grass shrimp were more sensitive to technical resmethrin and Scourge than the adult life stage. The results also demonstrate that synergized resmethrin is more toxic to P. pugio than the nonsynergized form, and that the presence of sediment decreases the toxicity of both resmethrin and Scourge.     

Equilibrium partitioning theory to predict the sediment toxicity of the anionic surfactant C(12)-2-LAS to Corophium volutator

The study of the effect of the sorption of linear alkylbenzene sulfonates (LAS) on the bioavailability to marine benthic organisms is essential to refine the environmental risk assessment of these compounds. According to the equilibrium partitioning theory (EqP), the effect concentration in water-only exposure will be similar to the effect concentration in the sediment pore water. In this work, sorption and desorption experiments with two marine sediments were carried out using the compound C₁₂-2-LAS. The effect of the sediment sorption on the toxicity of benthic organisms was studied in water-only and in sediment bioassays with the marine mud shrimp Corophium volutator. In addition, three common spiking methods were tested for its application in the toxicity tests, as well as the stability of the surfactant during the water-only and sediment-water test duration. LC50 values obtained from water-only exposure showed a good correspondence with the pore water concentrations calculated from the sorption and desorption isotherms in the spiked sediments.     

Toxicity and heavy metal contamination of surficial sediments from the Bay of Thessaloniki (Northwestern Aegean Sea) Greece

Surficial sediments (the fraction < 2000 microm) from the Bay of Thessaloniki, in the Northwestern Aegean Sea, Greece were examined for heavy metal and organic carbon contents, as well as for acute LUMIStox toxicity of pore waters (PWs), wet sediment elutriates (WSEs) and dry sediment elutriates (DSEs) obtained from the solid material remaining after PW extraction. WSEs where not toxic. EC20/50 values could be measured in some DSE and PW samples. In all sediment samples, the DSE toxicity was greater than the corresponding PW toxicity. Sediment concentrations of total and labile Cd, Pb, Cu, Cr, Zn, Mn, Ni and Fe were determined and evaluated in relation to sediment quality guidelines. Pollution levels ranged from low to high for certain metals. A misfit between sediment toxicities calculated from heavy metal concentrations and those biologically measured was observed. Toxicity values were in general poorly correlated with sediment's heavy metal or organic carbon content. As significant correlation was however found between the DSE toxicity with total Ni and labile Fe, as well as for the PW toxicity with total Ni, labile Fe and labile Cr.     

Bioaccumulation and toxicity of a cationic surfactant (DODMAC) in sediment dwelling freshwater invertebrates

Dimethyldioctadecylammonium chloride (DODMAC, CAS No. 107-64-2) is the principal active component of Di(hydrogenated tallow alkyl) dimethylammonium chloride (DHTDMAC, CAS No. 61789-80-8), a cationic surfactant formerly used principally in laundry fabric softeners. After discharge to water, DODMAC partitions strongly to sediment, therefore the assessment of the effects of DODMAC to benthic organisms is essential in any risk assessment. Chronic toxicity studies were conducted with Lumbriculus variegatus (Oligochaete), Tubifex tubifex (Oligochaete) and Caenorhabditis elegans (Nematode). NOECs were greater than 5738, 1515 and 1351 mg/kg dw, respectively, even for sub-lethal effects. Measurement of the route of uptake of DODMAC by L. variegatus demonstrated the relative importance of uptake via ingestion (86%) compared with direct contact with the sediment and via pore water (14%). The overall tendency of DODMAC to bioaccumulate, however, was low with measured accumulation factors of 0.22 and 0.78 for L. variegatus and T. tubifex, respectively.     

A review on the application of microbial toxicity tests for deriving sediment quality guidelines

The results of microbial toxicity tests are needed for the risk assessment of polluted sediments. In comparison with animals the anaerobic microorganisms are more tolerant to natural sediment conditions whereas they are more sensitive for a number of specific pollutants. Microbial toxicity tests from a literature search were classified in seven categories. Category A, B and C use polluted sediments and are applied for sediment monitoring. In category D, a pure chemical is added and the organisms and the test conditions were derived from sediment. Therefore this category can be used for setting sediment quality guidelines which protect sediment functions for the toxic effects of chemicals. In category E, organisms from a polluted site are separated from the sediment and are tested with pure chemicals. Organisms from a more polluted site can be more tolerant to a local pollutant. This is called pollution-induced community tolerance and can be used as evidence for the occurrence of toxic effects in a specific sediment. In category F pure chemicals are tested with a pure culture of microorganisms under sediment conditions. The results of category F tests can be combined with single species tests with animals and plants to obtain sediment quality guidelines sufficient for species protection. This can be compared with the sediment quality guidelines which protect sediment functions. When one of these quality guidelines is exceeded for a compound at a specific location a category E test can be used to determine whether the compound shows toxic effects in that sediment.     

Comparison of test procedures for sediment toxicity evaluation with Vibrio fischeri bacteria

The use of bacterial luminescence assays is particularly effective in the contaminated sediment evaluation. In the present study, a comparison was made of different baterial luminescence assays including acute toxicity of elutriates with Microtox® and LUMIStox®; chronic toxicity of elutriates with LUMIStox®; and, acute toxicity of solvent extracts with both the tests and the Solid Phase test with Microtox. The toxicity assay procedures were utilised for the Po River sediment toxicity evaluation, testing samples with different physical and chemical characteristics. The results evidentiated that each sediment test procedures provided indipendent and complementary ecotoxicological responces usefull for a sediment classification of the most polluted samples.     

Toxicity of the Mosquito Control Pesticide Scourge® to Adult and Larval Grass Shrimp (Palaemonetes pugio)

Abstract

This study investigated the toxicity of various concentrations of technical resmethrin and Scourge® on adult and larval Palaemonetes pugio, a common grass shrimp species. Two types of tests were conducted for each of the resmethrin formulations using adult and larval grass shrimp life stages, a 96-h static renewal aqueous test without sediment, and a 24-h static nonrenewal aqueous test with sediment. For resmethrin, the 96-h aqueous LC₅₀ value for adult shrimp was 0.53 μg/L (95% confidence interval (CI): 0.46–0.60 μg/L), and for larval shrimp was 0.35 μg/L (95% CI: 0.28–0.42 μg/L). In the presence of sediment, technical resmethrin produced a 24-h LC₅₀ value for adult shrimp of 5.44 μg/L (95% CI: 4.52–6.55 μg/L), and for larval shrimp of 2.15 μg/L (95% CI: 1.35–3.43 μg/L). For Scourge®, the 96-h aqueous LC₅₀ for adult shrimp was 2.08 μg/L (95% CI: 1.70–2.54 μg/L), and for larval shrimp was 0.36 μg/L (95% CI: 0.24–0.55 μg/L). The 24-h sediment test yielded an LC₅₀ value of 16.12 μg/L (95% CI: 14.79–17.57 μg/L) for adult shrimp, and 14.16 μg/L (95% CI: 12.21–16.43 μg/L) for larvae. Adjusted LC₅₀ values to reflect the 18% resmethrin concentration in Scourge® are 0.37 μg/L (adult), 0.07 μg/L (larvae) for the 96-h aqueous test, and 2.90 μg/L (adult), 2.6 μg/L (larvae) for the 24-h sediment test. Larval grass shrimp were more sensitive to technical resmethrin and Scourge® than the adult life stage. The results also demonstrate that synergized resmethrin is more toxic to P. pugio than the nonsynergized form, and that the presence of sediment decreases the toxicity of both resmethrin and Scourge®     

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.     

大型蚤标志酶用于城市二级出水毒性的评价

大型蚤是一种国际公认的标准实验生物,广泛应用于污水、地表水等水质毒性检测。毒性水平较低的城市二级出水,对大型蚤往往无急性毒性效应,而具有慢性毒性效应,但慢性毒性检测周期过长,因此探索一种更灵敏的指标,实现快速检测,对于控制二级出水的水质风险具有重要意义。本研究考察了大型蚤在短期暴露于城市二级出水条件下,其体内乙酰胆碱酯酶、超氧化物歧化酶、过氧化氢酶、ATP酶、羧酸酯酶和碱性磷酸酯酶的酶活变化特征,从中筛选出对二级出水毒性响应灵敏的标志酶指标。实验结果表明,碱性磷酸酯酶、过氧化氢酶对二级出水毒性响应相对较灵敏,具有成为标志酶的潜力,研究结果为城市二级出水生物毒性评价方法优化提供新的思路。     

A chronic bioassay with the estuarine amphipod Corophium volutator: test method description and confounding factors

Methods of conducting a chronic sediment toxicity test with the estuarine amphipod Corophium volutator are described. They consist of a 49-day exposure, after which mortality, growth and reproduction are determined. Pilot experiments were used to optimize test design parameters such as temperature, duration, feeding and refreshing regimes, and effects of indigenous organisms. By way of further validation, the present study focused on the effects of four different parameters: oxygen saturation, salinity, ammonium and nitrite. These confounding factors might play an important role especially if the test is used for risk assessment of field-contaminated sediments. It is concluded that the present experimental design is well suited for chronic sediment exposures with C. volutator. The test can be performed at a broad range of salinity values, provided that controls are performed at the same salinity. Results further demonstrate that with the endpoints growth and reproduction this chronic test procedure is a factor 7-18 more sensitive to ammonium and nitrate than the standardized acute bioassay (endpoint mortality).     

Limitations of reverse polyethylene samplers (RePES) for evaluating toxicity of field contaminated sediments

Passive samplers are used to measure dissolved nonionic organic contaminants (NOCs) in environmental media. More recently, reverse polyethylene samplers (RePES) have been used with spiked sediments to recreate interstitial water exposure concentrations and observed toxicity. In the present study, RePES were used with field contaminated sediments. The RePES was not capable of recreating the pattern of toxicity with the amphipod and mysid observed with intact field sediments. Decreased survival in the RePES exposures as compared to the whole sediment exposures was most likely caused by an overexposure to NOCs due to a lack of surrogate black carbon in the RePES system. As an alternative, aqueous phase studies were performed in which polyethylene was allowed to equilibrate with slurries of intact sediments for 3 weeks. Three weeks was found to be an insufficient amount of time for the polyethylene to equilibrate with the sediment. An additional study demonstrated 3 months was sufficient for lower contaminant concentrations, but might not be an adequate amount of time for more highly contaminated sediments. The aqueous phase transfer approach may be useful if equilibration is sufficiently long, although this length of time may be impractical for use in certain applications, such as toxicity identification evaluations (TIEs).     

The effects of engineered nanoparticles on survival, reproduction, and behaviour of freshwater snail, Physa acuta (Draparnaud, 1805)

Increasing uses of engineered nanoparticles (ENPs) in commercial products and industrial applications has eventually resulted to their releases into atmospheric, terrestrial, and aquatic environments. However, knowledge gaps in ENPs toxicity, fate, and behaviour currently limit our ability to quantify risk assessment of materials with nanoscale dimensions, and therefore, the extent of the resultant environmental impacts remains unknown. In the present study, we evaluated the effects of γ-alumina, α-alumina, modified TiO(2) (M-TiO(2)), and commercial TiO(2) (C-TiO(2)) ENPs on the survival, behaviour, and early life stages of the freshwater snail Physa acuta (Draparnaud). The toxicity evaluation was carried out after spiking commercial sand with ENPs concentrations of 0.005, 0.05, or 0.5 gk g(-1). Our findings suggest that increases of γ-alumina and α-alumina concentrations at sub-lethal level concentrations caused significant reduction in the embryo growth rate and embryo hatchability. In addition, these ENPs induced observable developmental deformities of the embryos. In addition, toxicity evaluations using acute 96-h and chronic 28-d tests showed exposure duration may be a significant factor in ENPs-induced toxicity. Therefore, long-term exposure of aquatic organisms to ENPs - potentially can alter certain ecological populations at different trophic levels - and may compromise the entire aquatic ecological functionality. The percentage hatchlings in test chambers containing 0.5 gk g(-1) γ-alumina and α-alumina concentration was 50% less to those observed in the controls. Our results suggest the embryonic growth and hatchability tests are useful endpoints in chronic sediment toxicity tests for determining the toxic thresholds of ENPs in sediment environment. Although no snail mortalities were observed during the static 96-h test containing sediment spiked with different concentrations of M-TiO(2), C-TiO(2), γ-alumina and α-alumina - the antioxidant enzymatic assay results indicated a significant change in antioxidant levels which altered peroxidation at 0.05 or 0.5 gk g(-1)concentrations for both γ-alumina and α-alumina.     

Influence of sediment composition on PAH toxicity using zebrafish (Danio rerio) and Japanese medaka (Oryzias latipes) embryo-larval assays

Due to hydrophobic and persistent properties, polycyclic aromatic hydrocarbons (PAHs) have a high capacity to accumulate in sediment. Sediment quality criteria, for the assessment of habitat quality and risk for aquatic life, include understanding the fate and effects of PAHs. In the context of European regulation (REACH and Water Framework Directive), the first objective was to assess the influence of sediment composition on the toxicity of two model PAHs, benzo[a]pyrene and fluoranthene using 10-day zebrafish embryo-larval assay. This procedure was undertaken with an artificial sediment in order to limit natural sediment variability. A suitable sediment composition might be then validated for zebrafish and proposed in a new OECD guideline for chemicals testing. Second, a comparative study of toxicity responses from this exposure protocol was then performed using another OECD species, the Japanese medaka. The potential toxicity of both PAHs was assessed through lethal (e.g., survival, hatching success) and sublethal endpoints (e.g., abnormalities, PMR, and EROD) measured at different developmental stages, adapted to the embryonic development time of both species. Regarding effects observed for both species, a suitable artificial sediment composition for PAH toxicity testing was set at 92.5 % dry weight (dw) silica of 0.2–0.5-mm grain size, 5 % dw kaolin clay without organic matter for zebrafish, and 2.5 % dw blond peat in more only for Japanese medaka. PAH bioavailability and toxicity were highly dependent on the fraction of organic matter in sediment and of the K _ow coefficients of the tested compounds. The biological responses observed were also dependent of the species under consideration. Japanese medaka embryos appeared more robust than zebrafish embryos for understanding the toxicity of PAHs following a sediment contact test, due to the longer exposure duration and lower sensitivity of sediment physical properties.     

Applying adult emergence as an endpoint in a post-exposure laboratory test using two midge species (Diptera: Chironomidae)

Several approaches have been used to evaluate biological impairment in aquatic ecosystems which can be categorized as either laboratory or field. In the recent years, the laboratory toxicity test approach has been extended to field exposures where ambient factors are allowed to influence the test response. Field exposures of laboratory test organisms require method modifications. In this paper, a novel in situ method is described which measures growth, survival and emergence of sediment inhabiting insects (Diptera: Chironomidae) that are used in standardized laboratory toxicity testing. Two standard chironomid species (Chironomus riparius and Chironomus tentans) were used to test the suitability of the approach and to compare the performance of the species. The larvae were transferred to the laboratory for emergence after 7 days in situ exposure which was compared to laboratory responses. Growth, survival and emergence were significantly lower in the in situ pre-exposure than in the laboratory. Also, emergence success was significantly lower in one reference sediment (LMR) than in the other test sediments in both in situ and the laboratory treatment. These lower response levels likely resulted from sediment characteristics and artifacts related to the exposure in the in situ chamber. Feeding and water quality within the exposure chamber appear to be factors that may differ markedly from the laboratory exposure and may affect organism responses. C. riparius developed (growth, emergence time) faster than C. tentans in all treatments, otherwise the species responded similarly. C. riparius may be a better alternative for the chronic in situ exposures because of shorter exposure times and reduced feeding requirements.