Assessment of lead toxicity to the marine bacterium, Vibrio fischeri, and to a heterogeneous population of microorganisms derived from the Pearl River in Jackson, Mississippi, USA.

Microorganisms are known to be excellent test organisms because of the relative ease for handling and suitability for analysis related to their small size, large number and convenient growing conditions. In this research, we tested the toxic effects of lead against a marine bacterium (Vibrio fischeri), and a heterogeneous population of bacteria derived from the Pearl River in Jackson, Mississippi. Using the level of bioluminescence in the Microtox Assay (V. fischeri), and the kinetics of dissolved oxygen uptake and growth (mixed bacterial population) as measures of toxicity, lead concentrations effecting a 50% reduction in these parameters (EC50) were determined as the toxic end-points. The activity quotients were also computed to determine the degrees of toxicity. Optical density (measure of growth) and oxygen uptake were measured over an extended period of time (20 h). EC50 values of 0.34 +/- 0.03, 3.10 +/- 0.01, and 3.80 +/- 0.02 mg/L were recorded for bioluminescence, growth, and oxygen uptake, respectively. As expected, the results indicated that the sensitivity to lead toxicity of V. fischeri was about one order of magnitude (10 times) greater than that of the mixed population of Pearl River microorganisms. Reductions in bioluminescence, growth, and oxygen uptake were directly correlated to lead concentrations, with toxic levels ranging from slightly toxic in lower concentrations to extremely toxic in higher concentrations. Upon 20 h of exposure, the times required to produce 50% reduction in dissolved oxygen uptake were (TD50S) 8.01 +/- 0.44, 9.60 +/- 0.46, 11.29 +/- 0.46, 13.03 +/- 0.57, 17.32 +/- 0.95, and 20.00 +/- 0.00 h in 0, 1, 2, 3, 4, 5, and 6 mg/L of lead, respectively, indicating a time-response relationship with respect to lead toxicity.     

Toxicity of mono- and diesters of o-phthalic esters to a crustacean, a green alga, and a bacterium

The degradation of phthalic acid diesters may lead to formation of o-phthalic acid and phthalic acid monoesters. The ecotoxic properties of the monoesters have never been systematically investigated, and concern has been raised that these degradation products may be more toxic than the diesters. Therefore, the aquatic toxicity of phthalic acid, six monoesters, and five diesters of o-phthalic acid was tested in three standardized toxicity tests using the bacteria Vibrio fischeri, the green algae Pseudokirchneriella subcapitata, and the crustacean Daphnia magna. The monoesters tested were monomethyl, monoethyl, monobutyl, monobenzyl, mono(2-ethylhexyl), and monodecyl phthalate, while the diesters tested were dimethyl, diethyl, dibutyl, butylbentyl, and di(2-ethylhexyl)phthalate, which were assumed to be below their water solubility. The median effective concentration (EC50) values for the three organisms ranged from 103 mg/L to >4.710 mg/L for phthalic acid, and corresponding values for the monoesters ranged from 2.3 mg/L (monodecyl phthalate in bacteria test) to 4,130 mg/L (monomethyl phthalate in bacteria test). Dimethyl and diethyl phthalate were found to be the least toxic of the diesters (EC50 26.2-377 mg/L), and the toxicity of the other diesters (butylbenzyl and dibutyl phthalate) ranged from 0.96 to 7.74 mg/L. In general, the phthalate monoesters (degradation products) were less toxic than the corresponding diesters (mother compounds).     

An algal toxicity database of organic toxicants derived by a closed-system technique

The current study presents the toxicity data of 90 organic compounds with various modes of actions to Pseudokirchneriella subcapitata. The assessment was conducted using a closed-system technique, and a biomass-type end point based on the cell density was employed. The above toxicity data were compared with test results from ciliate (Tetrahymena pyriformis), water flea (Daphnia magna), fish (Pimephales promelas), and luminescent bacteria (Photobacterium phosphoreum). Satisfactory correlation relationships between toxicity data from algae and other aquatic organisms were found (r2 = 0.66-0.82). Pseudokirchneriella subcapitata revealed considerably higher sensitivity to organic toxicants compared with other organisms. Benzenes, aldehydes, and alkanes also were highly selective to the test alga. In addition, the results show that conventional algal batch tests tend to underestimate the toxicity of organic compounds, except in the case of 4-chlorophenol. Toxicity observed from the closed-system test is approximately 2- to 380-fold higher than that estimated by conventional batch tests. Such a phenomenon can be found in nearly all organic compounds, regardless of the chemical's Henry's law constant. In the risk assessment of chemicals, following the European Union's practice, approximately 30% (7 of 23) of the cases may result in a more strict classification when the batch test is replaced by the closed-system test. More effort therefore is needed to revise the algal toxicity database using the closed-system test method.     

Ecotoxicological evaluation of wastewater treatment plant effluent discharges: a case study in Prato (Tuscany, Italy)

Textile wastewaters, which contain numerous chemicals such as dyes, surfactants, solvents, organic and inorganic salts, can cause severe pollution problems for the receiving freshwaters. The ecotoxicity of wastewaters in Prato, where there are about 14,000 textile and related factories, was investigated from 1996-1999 by means of bioassays. 147 samples of reclaimed wastewater were collected at the outlets of 4 centralized wastewater treatment plants. The acute and chronic toxicity of the effluents was measured with bioassays using three different target organisms: green algae (Pseudokirchneriella subcapitata), crustaceans (Daphnia magna) and bioluminescent bacteria (Vibrio fischeri). Toxicity was expressed as Effective Concentration 50 (EC50) and Toxic Units (TU). The results indicated that the effluents did not have significant acute toxicity: only 2.74% (EC50<100%, TU>1) of the 146 samples tested with crustaceans and 6.52% (EC50<50%, TU>2) of the 78 tested with bioluminescent bacteria showed toxic effects. With algae, slight chronic toxicity was found in 49.33% (mean EC50 value=86.56%, mean TU=1.16) of the 140 samples tested. The highest relative response was found with the algal assay using Pseudokirchneriella subcapitata: 49.33% of 140 samples showed chronic toxicity at 96 hours (EC50<100%).     

Ecotoxicity of quinoline and hydroxylated derivatives and their occurrence in groundwater of a tar-contaminated field site

In the groundwater of a timber impregnation site higher concentrations of hydroxylated quinolines compared to their parent compounds quinoline and isoquinoline were found. Studying the toxicity of parent compounds and metabolites, genotoxicity was found with metabolic activation in the SOS-Chromotest and Ames fluctuation test only for quinoline. An adverse effect on algae was observed only for the parent compounds quinoline and isoquinoline, while in the Daphnia magna immobilization assay most hydroxylated quinoline derivatives showed toxicity. The highest ecotoxic potential was observed in the Vibrio fischeri luminescence-inhibition assay. Comparing experimental EC50-values with QSAR predicted ones, for all compounds apart from isoquinoline and 2(1H)-quinolinone in the V. fischeri test baseline toxicity or polar nacrosis is indicated. In conclusion, the hydroxylation of quinoline leads to a detoxification of the genotoxic potential, while taken additive mixture toxicity and a safety factor into account parent compounds and metabolites are found of ecotoxicological relevance in the groundwater.     

Assessment of the toxicity of triasulfuron and its photoproducts using aquatic organisms

The toxicological effects of the sulfonylurea herbicide triasulfuron and its photoproducts were assessed on four aquatic organisms. Toxicity varied with tested organism and with triasulfuron irradiation time. Triasulfuron and its photoproducts had no significant effects on the crustacean (Cladocera) Daphnia magna (causing 50% effective concentration [EC50] [48 h] = 49 +/- 1 mg/L) and the marine bacteria Vibrio fischeri (EC50 [30 min] > 100 mg/L). In contrast, primary producers (the duckweed Lemna minor, the microalgae Pseudokirchneriella subcapitata, and Chlorella vulgaris) were very sensitive to triasulfuron (EC50s < 11 microg/L). For these organisms, triasulfuron photoproducts were less toxic than the parent compound but the residual toxicity observed still represented a potential environmental hazard.     

Ecotoxicity of a polycyclic aromatic hydrocarbon (PAH)-contaminated soil

Soil samples from a former cokery site polluted with polycyclic aromatic hydrocarbons (PAHs) were assessed for their toxicity to terrestrial and aquatic organisms and for their mutagenicity. The total concentration of the 16 PAHs listed as priority pollutants by the US Environmental Protection Agency (US-EPA) was 2634+/-241 mg/kgdw in soil samples. The toxicity of water-extractable pollutants from the contaminated soil samples was evaluated using acute (Vibrio fischeri; Microtox test, Daphnia magna) and chronic (Pseudokirchneriella subcapitata, Ceriodaphnia dubia) bioassays and the EC values were expressed as percentage water extract in the test media (v/v). Algal growth (EC50-3d=2.4+/-0.2% of the water extracts) and reproduction of C. dubia (EC50-7d=4.3+/-0.6%) were the most severely affected, compared to bacterial luminescence (EC50-30 min=12+/-3%) and daphnid viability (EC50-48 h=30+/-3%). The Ames and Mutatox tests indicated mutagenicity of water extracts, while no response was found with the umu test. The toxicity of the soil samples was assessed on the survival and reproduction of earthworms (Eisenia fetida) and collembolae (Folsomia candida), and on the germination and growth of higher plants (Lactuca sativa L.: lettuce and Brassica chinensis J.: Chinese cabbage). The EC50 values were expressed as percentage contaminated soil in ISO soil test medium (weight per weight-w/w) and indicated severe effects on reproduction of the collembola F. candida (EC50-28 d=5.7%) and the earthworm E. fetida (EC50-28 d=18% and EC50-56 d=8%, based on cocoon and juvenile production, respectively). Survival of collembolae was already affected at a low concentration of the contaminated soil (EC50-28 d=11%). The viability of juvenile earthworms was inhibited at much lower concentrations of the cokery soil (EC50-14 d=28%) than the viability of adults (EC50-14 d=74%). Only plant growth was inhibited (EC50-17d=26%) while germination was not. Chemical analyses of water extracts allowed us to identify inorganic water-extractable pollutants as responsible for toxicity on aquatic species, especially copper for effects on D. magna and C. dubia. The soil toxicity on collembolae and earthworms could be explained by 4 PAH congeners-fluorene, phenanthrene, pyrene, and fluoranthene. Yet, toxicity of the cokery soil as a whole was much lower than toxicity that could be deduced from the concentration of each congener in spiked soils, indicating that pollutants in the soil became less bioavailable with ageing.     

Effect of simulated sunlight on atrazine and metolachlor toxicity of surface waters.

Atrazine and metolachlor are the two most widely used herbicides in the United States; through non-point-source runoff both herbicides may cause toxicity to aquatic organisms. Toxicity changes were measured for atrazine and metolachlor in surface waters after exposure to simulated sunlight (0, 20, and 40 kJ/m2) using a Xenon Weather-Ometer. A Microtox toxicity test, using the marine luminescent bacterium Vibrio fischeri, was conducted on deionized, river, and bay water samples mixed with atrazine or metolachlor herbicide (12 mg/liter) after exposure to simulated sunlight. Microtox test (EC50%) results demonstrated that the toxicity decreased with increasing light intensity for both herbicides in river and bay water. These results also indicate that the toxicity of the bay water, with high concentrations of organic and suspended matter, was reduced, for both herbicides, compared with the toxicity of the river water, possibly through photodegradation of pesticides.     

砷与氟对三种环境生物的联合毒性研究

采用斑马鱼、大型水蚤及美丽猛水蚤对水中砷与氟的联合毒性进行了研究。用毒性单位法及相加指数法评价结果，３种生物的联合毒性实验都表明砷与氟在水环境中呈拮抗作用。     

Predicting acute zinc toxicity for Daphnia magna as a function of key water chemistry characteristics: development and validation of a biotic ligand model

The individual effect of different major cations (Ca2+, Mg2+, Na+, K+, and H+) on the acute toxicity of zinc to the waterflea Daphnia magna was investigated. The 48-h median effective concentration (EC50) in the baseline test medium (i.e., a standard medium with very low ion concentrations) was about 6 microM (Zn2+). An increase of Ca2+ (from 0.25 mM to 3 mM), Mg2+ (from 0.25 mM to 2 mM), and Na+ activity (from 0.077 mM to 13 mM) reduced zinc toxicity by a factor of 6.3, 2.1, and 3.1, respectively. No further toxicity reduction was observed when Ca2+ and Mg2+ activities exceeded 3.0 and 2.0 mM, respectively. Both K+ and H+ did not significantly alter zinc toxicity (expressed as Zn2+ activity). From these data, conditional stability constants for Ca2+ (log K = 3.24), Mg2+ (log K = 2.97), Na+ (log K = 2.16), and Zn2+ (log K = 5.31) were derived and incorporated into a biotic ligand model (BLM) predicting acute zinc toxicity to D. magna in surface waters with different water quality characteristics. Validation of the developed BLM using 17 media with different pH, hardness, and dissolved organic carbon (DOC) content resulted in a significant correlation coefficient (R2 = 0.76) between predicted and observed 48-h EC50. Eighty-eight percent of the predictions were within a factor of 1.3 of the observed 48-h EC50.     

Multiple computer-automated structure evaluation study of aquatic toxicity. III. Vibrio fischeri

An acute toxicity model was constructed on the basis of 901 chemicals tested for toxicity against the luminescent bacteria Vibrio fischeri (formerly Photobacterium phosphoreum, the Microtox test). The model was created using the Multiple Computer-Automated Structure Evaluation (M-CASE) program. The model can correctly predict acute toxicity for 92% of the compounds with an error averaging 0.55 log units per median effect concentration (EC50). The main toxicophores, corresponding to polar and nonpolar narcosis, and other types of reactive chemicals were identified.     

Ecotoxicity of mixtures of antibiotics used in aquacultures

More or less well-defined mixtures of antibiotics used in aquacultures may be distributed in the aquatic environment. Therefore, a systematic mixture ecotoxicity study was performed with the aquaculture antibiotics oxytetracycline, oxolinic acid, erythromycin, florfenicol, and flumequine. Test organisms were freshwater algae (Pseudokirchneriella subcapitata), activated sludge microorganisms, and luminescent bacteria (Vibrio fischeri). Design and statistical analysis of test results were based on isobolographic analysis. Synergistic effects were observed when combinations of erythromycin and oxytetracycline were tested on activated sludge microorganisms, and in these cases model predictions indicate independent action on the different bacterial species in the sludge. As predicted from the modes of action, concentration addition was evident when flumequine and oxolinic acid were mixed and tested on sludge bacteria. In the algae test, the combined toxicity of antibiotics could not be predicted based on knowledge of the modes of action of the individual compounds. Independent of the test species, our results gave examples of combined effects that were higher than predicted based on the assumption of concentration addition. This result underlines the need to consider the effects of mixtures of antibiotics on environmental organisms. The isobolographic method appears to be a suitable tool for this purpose, particularly for well-defined mixtures with few substances.     

环境生物测试技术对沉积物的毒性评价研究

作者采用５种环境生物测试技术对化工沉积物的毒性进行了评价，结果表明化工沉积物对卤虫（海洋生物）、美丽猛水蚤（河口生物）、大型水蚤（淡水生物）及斑马鱼都有一定毒性，其中斑马鱼胚胎仔鱼对化工沉积物的毒性最敏感。作者指出５种环境生物测试技术都可以用来评价沉积物的毒性。     

A predictive quantitative structure-activity relationship model for the photoinduced toxicity of polycyclic aromatic hydrocarbons to Daphnia magna with the use of factors for photosensitization and photomodification

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants that readily absorb environmentally relevant solar ultraviolet radiation. On absorption of a photon, photoinduced toxicity of PAHs is manifested through photosensitization and photomodification. Both of these processes occur under environmentally relevant levels of actinic radiation. An empirical quantitative structure-activity relationship model previously developed was explanatory of photoinduced toxicity of 16 PAHs in Lemna gibba (duckweed). This model was found to be predictive of toxicity to Vibrio fischeri. The L. gibba quantitative structure-activity relationship showed that a photosensitization factor and a photomodification factor could be combined to describe photoinduced toxicity. To further examine this model, we assessed whether it could be applied to Daphnia magna (water flea), a key bioindicator species in aquatic ecosystems. Toxicity was assessed as median effective concentration and median effective time for immobility. As with L. gibba and V. fischeri, neither the photosensitization factor nor the photomodification factor alone correlated to toxicity in D. magna. However, a photosensitization factor modified for D. magna exhibited a correlation to toxicity (r2 = 0.86), which was modestly improved when summed with a modified photomodification factor (r2 = 0.92). The greatest correlation was observed with median effective concentration data. This research provides evidence that models incorporating factors for photosensitization and photomodification have interspecies applicability.     

Ecological hazard assessment of major veterinary benzimidazoles: acute and chronic toxicities to aquatic microbes and invertebrates

Aquatic toxicities of six benzimidazole-based anthelmintics-namely, albendazole, thiabendazole, flubendazole, febantel, fenbendazole, and oxfendazole-were evaluated with a marine bacterium, Vibrio fischeri, and a freshwater invertebrate, Daphnia magna. Delayed and chronic toxicity tests using D. magna also were conducted for benzimidazoles with high acute toxicity. Vibrio fischeri was greater than 10-fold less sensitive to most of the benzimidazoles tested compared to daphnids. For D. magna, the most acutely toxic anthelmintic compound tested was fenbendazole (48-h median effective concentration [EC50s], 16.5 microg/L), followed by flubendazole (48-h EC50, 66.5 micro/L), albendazole (48-h EC50, 67.9 microg/L), febantel (48-h EC50, 216.5 microg/L), thiabendazole (48-h EC50, 843.6 microg/L), and oxfendazole (48-h EC50, 1,168.4 microg/L). The lipophilicity parameter, log Kow, explained the observed acute D. magna toxicity of the individual benzimidazoles (r = -0.91, p < 0.01). Delayed expression of toxicity observed for 21 d after 96-h exposure to fenbendazole and flubendazole was not notable, which might result from the relatively high elimination constants for the chemicals. With chronic exposure to fenbendazole, D. magna survival, reproduction, and growth were significantly impacted at 1.25 to 4.1 microg/L (p < 0.05). Hazard quotients estimated for fenbendazole, albendazole, flubendazole, and febantel were 2,770, 9.7, 4, and 1.2, respectively, suggesting a need for further investigation and a potential for environmental concerns, particularly with fenbendazole.     

水质毒性测定仪在水质急性毒性测定中的应用

目的 探讨TOXTRACER水质毒性测定仪在水质急性毒性测定中的应用。方法 采用荷兰生产的TOXTRACER水质毒性测定仪测定了不同条件下费氏弧菌(Vibrio fisheri)对HgCl2的抑制发光率,并对各种水样进行了抑制发光率和EC50值的测定。结果 0．20mg／L的HgCl2溶液在10、15、30min反应时间里的抑制发光率差异很小。0．10、0．20、0．40mg／L HgCl2溶液反应15min时的抑制发光率的变异系数分别为12．3％,2．6％,0．9％。在10—30min的反应时间里,HgCl2的浓度与其抑制发光率(％)之间线性关系均较好,EC50值(半数有效浓度)稳定。该仪器使用的费氏弧菌可连续使用约5h,它对HgCl2的敏感性优于国产冻干菌株(Photobacterium phosphoreum T3)。费氏弧菌的抑制发光率与0．2—40．0mg／L的As^2 ,0．5-5．0mg／L的Cr^3 ,0．05-0．40mg／L的Hg^2 呈线性关系。结论 该仪器可直接准确定量测定水质急性毒性。     

Importance of acclimation to environmentally relevant zinc concentrations on the sensitivity of Daphnia magna toward zinc

Daphnia magna was acclimated for six generations to an acclimation range of 0.02 to 74 microg/L of Zn2+. This range was determined by combining physicochemical water characteristics of European surface waters with total Zn concentrations in these waters in such a way that they resulted in minimal and maximal free (i.e., assumed bioavailable) Zn ion activities. No significant differences were found in acute Zn tolerance between the different acclimation concentrations: Average 48-h median effective concentration (EC50) values ranged from 608+/-94 to 713+/-249 microg/L of Zn2+. Also, no significant shifts in chronic tolerance were observed: Average 21-d EC50 (based on net reproductive rate) ranged from 91+/-20 to 124+/-22 microg/L of Zn2+. However, at test concentrations less than the 21-d EC50, acclimation significantly increased the reproductive capacity of the offspring produced. This indicates that metal acclimation is not necessarily accompanied by an increase in tolerance but also may manifest in other responses (e.g., reproduction rate). Organisms acclimated to a range from 6 to 22 microg/L of Zn2+ produced significantly more offspring than organisms acclimated to lower and higher Zn concentrations in test concentrations up to 50 microg/L of Zn2+. This range corresponds to a previously established optimal concentration range for D. magna. Bioconcentration factors indicated that Zn was actively regulated in the acclimation range tested.     

Modeling the concentration-response function of the herbicide dinoseb on Daphnia magna (survival time, reproduction) and Pseudokirchneriella subcapitata (growth rate)

Models describing dose-response relationships are becoming increasingly popular in ecotoxicology. They allow simple and thorough evaluations of toxicity test results, including inter- and extrapolations to concentrations or exposure times other than those tested. Simple parametric regression models are of particular interest because their parameters may be attributed mechanistic meanings and they can be applied without sophisticated mathematical and computational support. We recently proposed a four-parameter logistic regression model to fit the survival data of Daphnia magna under dinoseb stress. The model parameters are the maximum survival time, the minimum time required for an individual to die, effect concentration, EC(50), and a curve shape parameter. This model has now been applied to compare the lethality and reproduction toxicity of D. magna and the growth inhibition of Pseudokirchneriella subcapitata under dinoseb stress. It can be fitted adequately to all the measured data and the parameters can be attributed biological meanings in any of the three endpoints. A comparison of the modeled concentration-response functions of all three endpoints for dinoseb toxicity shows that the range of ECs with respect to both D. magna and algae is steep (a decrease of between 0.1 and 0.6 mg/L). The survival and reproduction of D. magna exhibit similar characteristic concentration-response functions and toxicities. The statistical no-effect concentration (SNEC) is 0.14 (survival) and 0.11 (reproduction)mg/L, respectively. On the other hand, algae seem to be less sensitive to dinoseb than D. magna (SNEC: 0.48 mg/L). However, further investigations of individual algae may lead to a more suitable comparison. We speculate that the four parameters of the model function can be related to specific properties of chemicals and organisms. Characterization of these properties would allow simple and appropriate estimation of the toxic effects of these chemicals.     

Toxic impact of effluents from petrochemical industry

The toxicity of effluents from a petrochemical industry center in southern Finland was tested by conducting bioassays on organisms from three different trophic levels. In fish tests, rainbow trout (Salmo gairdneri) were caged at the discharge site and simultaneously at a reference area. The only clear differences, among the measurements of 25 metabolic parameters, were observed in fish liver where activities of two detoxication enzymes were significantly increased in the exposed group. The water flea (Daphnia magna) was used both in acute (EC50) and long-term reproduction tests. No acute lethal toxicity was detected in any of the wastewater samples investigated. A combined effluent, however, caused a reduction in the reproduction rate with an EC50 of 3%. No mutagenic activity was observed with the Ames test (Salmonella typhimurium, strains TA 97, TA 98, and TA 100) in concentrated effluents, in sediment samples, or in liver samples from predator fish caught from the discharge site.