Study on the cytotoxicity of microcystin-LR on cultured cells

The toxicity of purified blue-green algal toxin, microcystin-LR, on permanent cell lines KB, NIH/3T3, H-4-II-E, HeLa, Vero, Hep G2, Caco-2 and HL-60 was studied. Assessment of cell viability using colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays indicated that purified microcystin-LR induced toxic effect on KB and H-4-II-E cell lines after 96 h incubation at toxin concentrations greater than 18.75 microg/ml. KB cell line was selected for further study when reproducibility, consistency and sensitivity were considered. Significant amounts of lactate dehydrogenase (LDH) were released from KB cells when incubation durations were 72 and 96 h with toxin concentrations of 18.75 microg/ml and higher. Although previous studies suggested that microcystin-LR had no cytotoxic effect on permanent cell lines, LDH release assay performed on KB cells indicated that exposure to microcystin-LR could result in damage to the cell membrane.     

Biomarker responses and reproductive toxicity of the effluent from a Chinese large sewage treatment plant in Japanese medaka (Oryzias latipes)

The present study was conducted to assess the potential toxicity of the effluent from a large sewage treatment plant (GBD-STP) in Beijing. Japanese medakas (Oryzias latipes) at reproduction active period were exposed to a serial of graded concentrations of the effluent or 100 ng l-1 of 17-alpha-ethinylestradiol (EE2, positive control). Growth, gonadosomatic index (GSI), hepatosomatic index (HSI), reproductive success, induction potency of vitellogenin (VTG) in male fish and that of 7-ethoxyresorufin-o-deethylase activity (EROD) in male fish liver were used as test endpoints. The growth suppression of fish was observed in a dose-dependent manner, resulting in significant differences in both body length and body weight of medaka above 5% effluent. This effluent can inhibit the growth of gonad of medakas and are more sensitive to male than to female. At exposure concentration of 40% and higher, there was an unexpected decrease of HSI values, which may be resulted from sub-lethal toxicity of effluent to fish liver. VTG of plasma in males were induced in all exposure concentration levels, but not in a dose-dependent manner. The concentration of 5% effluent would be the lowest observed adverse effect level (LOAEL) affecting reproductive success when examining fertile individuals, fecundity and fertilization rate. The overt CYP1A response and higher reproductive toxicity may be indicative of low process efficiency of this STP.     

Effects of the insect growth regulator (S)-methoprene on survival and reproduction of the freshwater cladoceran Moina macrocopa

This study examines the acute and chronic effects of the insecticide (S)-methoprene on the survival and reproduction of the freshwater cladoceran Moina macrocopa. In laboratory toxicity tests, the 24- and 48-h LC50 values for (S)-methoprene were 0.51 and 0.34 mg litre(-1), respectively. Survival, longevity and fecundity were reduced at 0.05 mg litre(-1) and higher concentrations. At 0.005 and 0.01 mg litre(-1), longevity and fecundity increased slightly as compared to control animals. If environmental concentrations of methoprene do not exceed 0.05 mg litre(-1), as is generally the case, application of this insecticide is unlikely to induce detrimental effects on natural cladoceran populations. The stimulatory effect of very low levels of methoprene on reproductive performance is consistent with the hypothesis of a regulatory role of juvenile hormone-like compounds in crustacean reproduction.     

Survival, growth and reproduction of Daphnia carinata (Crustacea: Cladocera) exposed to chronic cadmium stress at different food (Chlorella) levels

In nature, organisms have to respond to a diversity of factors acting simultaneously. The present investigation was conducted to study whether changes in food (Chlorella) levels could modify the chronic toxicity of cadmium on the various life-history parameters, such as survivorship, longevity, life expectancy, fecundity, age at first reproduction, R(0), T, r and growth rates of the cladoceran Daphnia carinata. The study indicated that at low food levels (0.5 x 10(6) cells ml(-1) Chlorella), cadmium concentrations in the range of 27-162 microg litre(-1) reduced these life-history parameters by 50% (EC(50)). At medium food levels (1.5 x 10(6) ml(-1) Chlorella) the EC(50) of cadmium was in the range of 51-127 microg litre(-1). At high food levels (4.5 x 10(6) cells ml(-1) Chlorella), the toxic effect of cadmium was greatly reduced. The decreases in survival, growth and reproduction of D. carinata at high cadmium-low food levels affected the fitness parameter 'r'. The study emphasises the need to include reproductive parameters other than mere survival in toxicity bioassays. The study also stresses the need to incorporate in laboratory tests other relevant factors that might modify pollutant toxicity.     

Impact of Parthenium weeds on earthworms (Eudrilus eugeniae) during vermicomposting

The aim of this work is to evaluate the effect of Parthenium-mediated compost on Eudrilus eugeniae during the process of vermicomposting. Nine different concentrations of Parthenium hysterophorus and cow dung mixtures were used to assess toxicity. The earthworms’ growth, fecundity and antioxidant enzyme levels were analysed every 15 days. The antioxidant activities of enzymes [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], considered as biomarkers, indicate the biochemical and oxidative stresses due to the toxin from Parthenium weeds. The earthworms’ growth, biomass gain, cocoon production and antioxidant enzymes were in a low level in a high concentration of P. hysterophorus (without cow dung). The results clearly indicated that appropriate mixing of P. hysterophorus quantity is an essential factor for the survival of earthworms without causing any harm.     

Behavioural and feeding responses of Echinogammarus meridionalis (Crustacea, Amphipoda) to acid mine drainage

As a result of mining activities the exposure of metal sulphides to oxidation takes place with consequent release of acid mine drainage (AMD). Biomonitoring instruments have been proven to have the best deterrent effect upon polluters. A new approach in online biomonitoring, with the Multispecies Freshwater Biomonitortrade mark (MFB), was developed combining behavioural and feeding responses of the Portuguese indigenous benthic shredder, Echinogammarus meridionalis (Pinkster, 1973) (Crustacea, Amphipoda). These endpoints, along with mortality, were measured and analyzed for a gradient of sublethal doses of AMD. Original river water was used as the control and three doses of treatments were attained by adding increasing volumes of AMD to the control. The increase in AMD concentration and concomitant decrease in pH and increase in the concentration of most metals was followed by an overall increase of the mortality, decrease of locomotion/feeding activity and inhibition of the feeding rate. Mortality was observed in the two highest concentrations of AMD. Significant decrease in average locomotion/feeding activity took place in the second treatment. Although an inhibition of feeding was observed along the gradient of AMD concentration only in the highest concentration the feeding rate was significantly reduced.     

Effects of mercury on behavior and performance of northern two-lined salamanders (Eurycea bislineata)

Mercury (Hg) causes a range of deleterious effects in wildlife, but little is known about its effects on amphibians. Our objective was to determine whether Hg affects performance and behavior in two-lined salamanders (Eurycea bislineata). We collected salamanders from Hg-contaminated and reference sites and assessed speed, responsiveness, and prey capture ability. Mercury concentrations were >17× higher in salamanders from the contaminated sites and were among the highest documented in amphibians. In the first, but not in the second, locomotion trial, we found a significant effect of Hg on speed and responsiveness. In the prey capture experiment, reference salamanders ate approximately twice as many prey items as the contaminated salamanders. Together, our results suggest that sublethal Hg concentrations may negatively affect salamanders by reducing their ability to successfully execute tasks critical to survival. Future work is warranted to determine whether Hg has other sublethal effects on salamanders and whether other amphibians are similarly affected.     

Energy resource reallocation in Daphnia schodleri (Anomopoda: Daphniidae) reproduction induced by exposure to hexavalent chromium

Cladocerans are able to store energy in the form of macromolecules such as proteins, lipids and carbohydrates. However, their ability to do so depends to a large extent on their physiological and reproductive condition as well as their age. These macromolecules constitute the total caloric reserves of the body and may be modified as a result of environmental stress conditions by either physical or nutritional factors or by exposure to toxic compounds. Neonates of the freshwater cladoceran Daphnia schodleri were exposed to 0.0064, 0.032 and 0.064 mg L⁻¹ of hexavalent chromium - Cr(VI) - up to age 5, 7, 14, 21 or 28 d. Survival and fecundity of parthenogenetic females was evaluated for 28 d (maximum exposure period). Body length and maximum width were determined in specimens of all ages. Caloric content was evaluated in progeny and females within each age group. Female survival was significantly lower with exposure to 0.064 mg L⁻¹ of Cr(VI) than with other treatments, and negative effects on reproduction were observed from the lowest concentration on. Largest adult size was attained by control specimens towards the end of the observation period, but no significant differences in neonate size occurred between concentrations. Energy content decreased in adults as toxicant concentration increased, but these females allocated a larger amount of energy reserves to their progeny as a possible compensation aimed at ensuring neonate survival in an adverse environment due to toxicant presence. Sublethal concentrations of Cr(VI) modified energy content in D. schodleri, affecting to varying degrees its survival, growth and reproduction.     

Effects of different food (Chlorella) concentrations on the chronic toxicity of cadmium to survivorship, growth and reproduction of Echinisca triserialis (Crustacea: Cladocera)

Extraneous factors have been shown to greatly modify pollutant stress. The present study was conducted with the objective of determining whether differences in food levels could modify chronic toxicity of cadmium to the various life-history parameters of the cladoceran Echinisca triserialis. Laboratory experiments were conducted on a sublethal range of cadmium (0, 2.5, 5.0, 10 and 20 microg litre(-1)) on life-history parameters such as survivorship, longevity, life expectancy, age at first reproduction, total fecundity, neonate size, net reproductive rate (R0), generation time (T), intrinsic rate of natural increase (r) and growth of Echinisca triserialis in relation to different food (Chlorella) levels of 0.5 (low), 1.5 (medium) and 4.5 (high) x 10(6) cells ml(-1). Cadmium levels of 10 microg litre(-1) and above, and low food levels, had a profound effect in decreasing the magnitude of all parameters studied. EC50 levels computed for life-history parameters, such as longevity, life expectancy at birth, total fecundity, R0 and T, were in the range of 2 to 21 microg litre(-1) cadmium, and this is indicative of extreme adverse effects on the population dynamics of E. triserialis when exposed to low food levels. At high food levels the EC50 was not achieved for cadmium in the toxicant range studied. The significance of these results is discussed in relation to field conditions.     

Chronic toxicity of tributyltin to development and reproduction of the European freshwater snail Lymnaea stagnalis (L.)

Chronic toxicity, growth and reproduction were measured in the freshwater gastropod Lymnaea stagnalis exposed to waterborne bis(tri-n-butyltin) oxide (TBTO) over a range of four nominal concentrations (0-10microg TBTl(-1)). Egg development was completely inhibited at 10microg TBTl(-1), whilst abnormal embryonic development was observed at 1microg TBTl(-1). For the solvent control and the 0.01microg TBTl(-1)treatment group, normal development of L. stagnalis was observed. Survivorship of hatchlings was significantly reduced by TBT at 1microgl(-1) while inhibition of shell growth of L. stagnalis was also observed at this concentration. The data were used to determine intrinsic growth rates (r) using two theoretical approaches (the Euler-Lotka equation and a Leslie Matrix). Both approaches showed that survival, fecundity and population growth rate were reduced at 1microg TBTl(-1). Interestingly, at 0.01microg TBTl(-1) snails showed a higher fecundity and growth rate than in the solvent control. The TBT concentration at which the r would equal zero (ECr(0)) and the population NOEC (No Observed Effect Concentration) were estimated. The population NOEC was defined as either the lower 95% confidence or lower 95% pointwise percentile limit of the ECr(0). Values obtained using the two different approaches were similar and thus a geometric mean was calculated to obtain a final representative population NOEC value for L. stagnalis of 2745ng TBTl(-1). The present data together with chronic toxicity TBT data for freshwater organisms, obtained from peer-reviewed literature, were used to construct a species sensitivity distribution (SSD). A predicted no effect concentration was then derived from the SSD (hazardous concentration at 5%, i.e., HC5 or 95% protection level). This SSD was compared with the SSD derived from saltwater species datasets. The HC5 value for saltwater species (3.55ng TBTl(-1); lower confidence limit: 1.93ng TBTl(-1)) was significantly lower than that for freshwater species (30.13ng TBTl(-1); lower confidence limit: 9.23ng TBTl(-1)), indicating that saltwater species are probably more susceptible to TBT than their freshwater counterparts.     

Chronic toxicity of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) in soil determined using the earthworm (Eisenia andrei) reproduction test

The sublethal and chronic effects of the environmental contaminant and explosive octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) in artificial soil were assessed using the earthworm (Eisenia andrei). Based on various reproduction parameters (total and hatched number of cocoons, number of juveniles and their biomass), fecundity was reduced at the different concentrations of HMX tested (from 280.0 +/- 12.3 to 2502.9 +/- 230.0 mg kg-1 dry soil) in spiked artificial soil (LOEC: 280.0 +/- 12.3 mg kg-1 dry soil). The growth of adult E. andrei was also reduced at the different concentrations tested, though no mortality occurred, even at the highest tested concentrations. The number of juveniles produced was correlated with the number of total and hatched cocoons, and the biomass of juveniles was correlated with the number of cocoons. Pooled results of these and earlier studies on explosives (TNT, RDX) using the E. andrei reproduction test confirm that effects of HMX on cocoon production are indicative of some reproductive consequences (number of juvenile and their biomass), whereas adult growth, in general, does not correlate strongly with change in reproduction capacity.     

Effects of a dark septate endophyte (DSE) on growth,cadmium content,and physiology in maize under cadmium stress

Dark septate endophytes (DSE) are widely distributed in plant roots grown in stressful habitats, especially in heavy metal-polluted soils. But little is known about the physiological interactions between DSE and plants under heavy metal stress. In the present study, the growth, Cd content, and physiological response of Zea mays L. to a root-colonized DSE, Exophiala pisciphila, were analyzed under Cd stress (0, 5, 10, 20, and 40 mg/kg) in a sand culture experiment. Under high Cd (10, 20, and 40 mg/kg) stress, the DSE colonization in roots increased the maize growth, kept more Cd in roots, and decreased Cd content in shoots. The DSE colonization improved the photosynthesis and induced notable changes on phytohormones but had no significant effect on the antioxidant capability in the maize leaves. Moreover, there were significant positive correlations between the gibberellic acid (GA) content and transpiration rate, zeatin riboside (ZR) content, and photosynthetic rate in maize leaves. These results indicated that the DSE’s ability to promote plant growth was related to a decrease on Cd content and the regulation on phytohormone balance and photosynthetic activities in maize leaves.

     

Effects of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in zebrafish: general and reproductive toxicity

Mixed-sex populations of young adult zebrafish (~2-month-old) were exposed to measured RDX concentrations of 0, 1 or 9.6 ppm for up to 12 weeks followed by a 15-day rearing period in untreated water. RDX caused high mortality at 9.6 ppm, with most deaths occurring within the first 8 weeks of exposure. RDX at 9.6 ppm caused lower body weights at 4 and 8 weeks of exposure; and at 1 ppm, lower body weight was observed only at 4 weeks. Fish length was not affected by treatment at any time during the exposure period. The bioconcentration factor for RDX seemed to be influenced by time of exposure but not by water RDX concentration; its overall values were 1.01+/-0.13, 0.91+/-0.06 and 2.23+/-0.04 at 4, 8 weeks and 12 weeks, respectively. RDX was not detected in fish collected after the 15-day recovery period. In a separate experiment, adult females and males were separately exposed to RDX at measured concentrations of 0, 0.5 and 3.2 ppm for a period of 6 weeks. Reproductive performance was evaluated by biweekly breeding of the fish and measuring packed-egg volume (PEV) as index of fecundity. At 0.5 ppm, RDX caused elevated PEV levels relative to the control value at 2 weeks but not at 4 or 6 weeks, whereas no significant effects were noted at 3.2 ppm. Egg fertilization and embryo hatching rates were not affected by RDX at any of the concentrations tested. In conclusion, RDX at sublethal concentrations causes short-term negative effects on growth and, at 0.5 ppm, positive effects on fecundity.     

Ecosystem engineering potential of the gastropod Terebralia palustris (Linnaeus, 1767) in mangrove wastewater wetlands - A controlled mesocosm experiment

The effect of different sewage concentrations (0, 20, 60 and 100%), vegetation (Bare, Avicennia marina or Rhizophora mucronata) and immersion periods (immersion/emersion period of 12/12 h or 3/3 days just for 100%) conditions were studied for 6 months on survival and growth rates of Terebralia palustris (Linnaeus, 1767). Gastropods' activity and ecosystem engineering preformed at bare and A. marina planted cells and 3 sewage conditions (0, 20 and 60%) were determined. Survival rates were higher than 70% in all treatments. Growth rate decreased significantly with increasing sewage concentrations (mainly at unplanted conditions) and longer immersion periods. A complete shift (from immersion to emersion periods) and a significant decrease in mobility and consequently its engineer potential, due to sewage contamination, lead to a 3-4 fold decrease in the amount of sediment disturbed. Sewage contamination, primary producers' abundance and environmental conditions may have influenced the gastropods survival, growth and its ecosystem engineering potential.     

Effects of a surfactant (FFD-6) on Scenedesmus morphology and growth under different nutrient conditions

Surfactants are man-made compounds that are meanwhile omnipresent in the environment, but environmental concentrations of surfactants are such that they are thought to have little risk for aquatic systems. The major anionic surfactants currently on the global market are linear alkylbenzene sulfonates (LAS), a class where the commercially available FFD-6 belongs to. The hypothesis was tested that sublethal effects of FFD-6, i.e. the morphological effect of colony formation in the common test alga Scenedesmus obliquus, occurs at a concentration lower than the no-observed-effect concentrations for endpoints commonly used in regulatory toxicity testing with algae. The surfactant FFD-6 induced colonies in Scenedesmus at concentrations a few orders of magnitude lower (i.e. between 0.001 and 0.01 g l-1) than at which growth inhibition was observed (i.e. between 1 and 10g l -1). Growth rates were lowest for Scenedesmus grown in P-limited medium, intermediate for algae reared in N-limited medium and highest for algae cultured in non-limited standard medium. Growth inhibition due to FFD-6 was similar for non-limited and nutrient-limited Scenedesmus, but colony formation was stronger in non-limited Scenedesmus than in nutrient limited cultures. The colony inducing effect of the surfactant FFD-6 on Scenedesmus occurs at much lower concentrations than growth inhibition and might affect species interactions, the survival of species and the energy flow along the food chain.     

Mortality, growth, swimming activity and gill morphology of brook trout (Salvelinus fontinalis) and Atlantic salmon (Salmo salar) exposed to low pH with and without aluminum

Measurements were made of mortality, growth, swimming activity, and gill morphology of young-of-the-year brook trout (Salvelinus fontinalis) and Atlantic salmon (Salmo salar), exposed for 30 days to pH 5.6 +/- 0.5 (mean +/- 1 standard deviation) with and without addition of 107 +/- 57 microg liter(-1) exchangeable (labile monomeric) aluminum. The experiment was conducted in artificial stream channels adjacent to a natural stream and subject to daily and seasonal changes in temperature, light, and chemical conditions. There were no differences in survival or growth for brook trout in any treatment; Atlantic salmon survival and growth were significantly decreased in the acid + Al treatment. Scanning electron microscopy showed no damage to gills of either species in the acid treatments, but the acid + Al treatment caused slight swelling of brook trout gills near the filament tips and significant swelling and fusion of secondary lamellae of Atlantic salmon gills. The acid treatment increased swimming activity in brook trout, but both the acid and acid + Al treatments reduced activity in Atlantic salmon.     

Evaluation of Acacia nilotica (L.) del. and Casuarina equisetifolia forst. for tolerance and growth on microbially treated dyestuff wastewater

Two tree species, Acacia nilotica (L.) Del. and Casuarina equisetifolia Forst., were tested for their tolerance and growth on dyestuff wastewater containing phenol, aniline, and methyl violet. The wastewater was treated microbially by using a culture of Pseudomonas alcaligenes in a fixed-film bioreactor. The plants were watered with untreated and treated wastewater and tap water (control) at the rate of 2 litre per plant per week. A. nilotica exhibited 100% survival with both untreated and treated wastewater, and growth (increase in height) was comparable to that of control plants. However, growth of C. equisetifolia was adversely affected. It exhibited 100% survival with treated wastewater but only 87% survival with untreated waste-water and the percentage increase in height was less with both treated and untreated wastewater. There was no effect on soil except for an increase in chloride content.     

光反应器中UV/Fenton光降解湖水中微囊藻毒素的研究

在光催化反应器中,采用UV/Fenton光催化氧化技术,对湖水中微囊藻毒素的光催化降解过程中各影响因子分析表明:微囊藻毒素光催化氧化降解率受光照强度、氧化剂种类及浓度、溶液pH和不同光催化体系等多因素的影响.光促催化氧化体系对MC-LR藻毒素降解具有显著的作用,在紫外光与氧化剂的协同作用下,紫外光照强度越高,越易于促进MC-LR藻毒素快速降解,降解率可到达80%以上.     

Effect of certain chemicals on the reproduction of medaka (Oryzias latipes)

In order to understand the effects of estrogenic chemicals on fish reproduction, we exposed male medaka (Oryzias latipes) to a natural estrogen [17 beta-estradiol (17 beta-E2)] and three estrogenic chemicals [bisphenol-A, nonylphenol (NP) and di(2-ethylhexyl)phthalate (DEHP)]. After two weeks' exposure, one male medaka was kept together with two female medaka for spawning, and the number of eggs and hatchings were compared to those of a negative control group. The results indicated that exposure to 17 beta-E2 caused a significant decrease in the number of eggs and hatchings as compared to the negative control group at and above 3 nmol/l. Also, the highest concentrations of bisphenol-A and NP caused a decrease in the number of hatchings, but no decrease in hatchings was observed in DEHP treatments. In the treatment using these chemicals the decrease in egg numbers was not so much as in hatching numbers. When compared to other in vitro studies, concentrations observed to have adverse effects on reproduction in this study are generally lower. In addition, it was suggested that physical alterations, such as an induction of plasma vitellogenin, were caused at much lower concentrations than those at which a decline in reproductivity was actually induced.     

Effects of thiobencarb herbicide to an alga (Nannochloris oculata) and the cladoceran (Daphnia magna)

Chronic toxicity studies were conducted with an algae (Nannochloris oculata) and the cladoceran (Daphnia magna) to determine their relative sensitivities to the thiocarbamate herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate). Most of the algal populations were initially affected by exposure to the herbicide. Thiobencarb concentrations higher than 0.5 mg/L significantly reduced algal densities after 24-h exposure. The 24-h static EC50 in D. magna was 3.01 mg/L. The sublethal effects of 0.3, 0.37, 0.5, 0.75, and 1.5 mg/L of thiobencarb concentrations on the survival, reproduction, and growth of D. magna were monitored for 21 days. The parameters used to determined the effect of the herbicide 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 was significantly reduced at thiobencarb concentrations of 0.30 mg/L and higher while survival was affected after exposure to 0.75 and 1.5 mg/L of the pesticide. The r value decreased with increasing concentrations of thiobencarb. Growth, as measured by body length, was depressed significantly after exposure to all herbicide concentrations tested.