Multi-generation cadmium acclimation and tolerance in Daphnia magna Straus

The cladoceran Daphnia magna was acclimated for seven generations to cadmium concentrations ranging from 0 (control) to 250 microg/l Cd (corresponding to a free ion activity of 4.60 nM Cd2+). Acute and chronic cadmium tolerance as well as cadmium accumulation were monitored as a function of acclimation time. After two to three generations of acclimation to concentrations ranging from 0.23 to 1.11 nM Cd2+ increases in acute tolerance were maximal (factor 7.2) and significant. Acclimation for seven generations to the same acclimation concentrations did result in an increased chronic cadmium tolerance (21 days EC50 values increased). Organisms acclimated to 1.93 nM Cd2+ were equally or more sensitive than non-acclimated daphnids in acute and chronic toxicity tests. Cadmium contents in D. magna increased significantly as a function of the acclimation concentration. Maximum body burdens of 236+/-30 microg Cd/g dry weight were measured in organisms exposed to 4.60 nM Cd2+, but detoxification mechanisms were only successful up to 82+/-20 microg Cd/g dry weight as this concentration did not cause major decreases in survival and reproduction in chronic toxicity tests. As the potential positive effect of acclimation on cadmium tolerance disappeared with successive acclimation generations and increasing acclimation concentrations, it is concluded that multi-generation acclimation studies are important for the evaluation of the long-term effects of environmental toxicants.     

Cadmium-tolerant bacteria induce metal stress tolerance in cereals

Cadmium usually hampers plant growth, but bacterial inoculation may improve stress tolerance in plants to Cd by involving various mechanisms. The objective was to characterize and identify bacteria that improve plant growth under Cd stress and reduce Cd uptake. Cadmium-tolerant bacteria were isolated from rhizosphere soil, which was irrigated with tannery effluent, and six strains were selected as highly tolerant to Cd, showing minimum inhibitory concentration as 500 mg L^?1 or 4.45 mmol L^?1. These strains were identified by 16S rRNA gene analysis and functional analysis in regard to plant growth promotion characteristics. To determine their effect on cereal growth under Cd stress, seeds were inoculated with these strains individually and grown in soil contaminated with three Cd levels (0, 40 and 80 mg kg^?1). Biomass production, relative water content (RWC), electrolyte leakage (ELL) and tissue Cd concentration were measured. Biomass of both cereals was inhibited strongly when exposed to Cd; however, bacterial inoculation significantly reduced the suppressive effect of Cd on cereal growth and physiology. The bacterial isolates belonged to the genera Klebsiella, Stenotrophomonas, Bacillus and Serratia. Maize was more sensitive than wheat to Cd. Klebsiella sp. strain CIK-502 had the most pronounced effects in promoting maize and wheat growth and lowering Cd uptake under Cd stress.     

Characterizing the toxicity of pulsed selenium exposure to Daphnia magna

The acute toxicity of selenium (Se) to aquatic biota has been studied extensively for decades. However, most studies have used a constant concentration aqueous exposure of Se to an invertebrate species. Since constant concentration exposure of toxicants to invertebrates is unusual in the environment, episodic exposure or pulsed exposures may represent true risk to aquatic biota more accurately. This research was designed to characterize the toxicity effects of pulsed Se exposure to Daphnia magna. Selenium exposure was varied during a 21-d chronic toxicity test to examine the effects of exposure concentration, duration, and recovery on survival, growth, and reproduction of D. magna. While D. magna did not die during exposures, latent mortality was observed. Latent mortality increased with exposure concentration and duration. Hence, standard toxicity test using continuous exposures would underestimate Se toxicity. Risk assessment method using results of continuous exposure would underestimate risk of Se to biota. For double-pulse exposures, cumulative mortality on day 21 was higher when time interval between pulses was shorter. With the same total exposure time, continuous exposure caused higher toxicity than did pulsed exposures due to recovery and tolerance development in D. magna after earlier pulses. Growth and reproduction of surviving D. magna were not affected by pulsed Se exposure due to recovery of D. magna after removal of the pulses. Based on these results, risk assessment for Se should take latent effects and the effect of recovery in to account.     

Cadmium tolerance in six poplar species

Selection of poplar species with greater Cd tolerance and exploiting the physiological mechanisms involved in Cd tolerance are crucial for application of these species to phyto-remediation. The aim of this study is to investigate variation in Cd tolerance among the six poplar species and its underlying physiological mechanisms. Cuttings of six Populus species were cultivated for 10 weeks before exposure to either 0 or 200 μM CdSO₄ for 20 days. Gas exchange in mature leaves was determined by a portable photosynthesis system. Cd concentrations in tissues were analyzed by a flame atomic absorbance spectrometry. Subsequently, Cd amount per plant, bio-concentration factor (BCF) and translocation factor (T _f) were calculated. Nonenzymatic compounds and activities of antioxidative enzymes in tissues were analyzed spectrophotometrically. Cd exposure caused decline in photosynthesis in four poplar species including Populus cathayana (zhonghua 1). Among the six species, P. cathayana (zhonghua 1) displayed the highest Cd concentrations in tissues, the largest Cd amount in aerial parts, the highest BCF in aerial parts and T _f under Cd exposure. Under Cd stress, increases in total soluble sugars in roots but decreases in starch in roots, wood, and leaves of P. cathayana (zhonghua 1) were found. Induced O ₂ ^?? and H₂O₂ production in roots and leaves, and increases in free proline, soluble phenolics, and activities of antioxidative enzymes were observed in P. cathayana (zhonghua 1). Based on results of this pot experiment, it is concluded that P. cathayana (zhonghua 1) is superior to other five species for Cd phyto-remediation, and its well-coordinated physiological changes under Cd exposure confer the great Cd tolerance of this species.     

Cadmium stress in rice: toxic effects,tolerance mechanisms,and management: a critical review

Cadmium (Cd) is one of the main pollutants in paddy fields, and its accumulation in rice (Oryza sativa L.) and subsequent transfer to food chain is a global environmental issue. This paper reviews the toxic effects, tolerance mechanisms, and management of Cd in a rice paddy. Cadmium toxicity decreases seed germination, growth, mineral nutrients, photosynthesis, and grain yield. It also causes oxidative stress and genotoxicity in rice. Plant response to Cd toxicity varies with cultivars, growth condition, and duration of Cd exposure. Under Cd stress, stimulation of antioxidant defense system, osmoregulation, ion homeostasis, and over production of signaling molecules are important tolerance mechanisms in rice. Several strategies have been proposed for the management of Cd-contaminated paddy soils. One such approach is the exogenous application of hormones, osmolytes, and signaling molecules. Moreover, Cd uptake and toxicity in rice can be decreased by proper application of essential nutrients such as nitrogen, zinc, iron, and selenium in Cd-contaminated soils. In addition, several inorganic (liming and silicon) and organic (compost and biochar) amendments have been applied in the soils to reduce Cd stress in rice. Selection of low Cd-accumulating rice cultivars, crop rotation, water management, and exogenous application of microbes could be a reasonable approach to alleviate Cd toxicity in rice. To draw a sound conclusion, long-term field trials are still required, including risks and benefit analysis for various management strategies.     

Inter- and intra-species variation in acute zinc tolerance of field-collected cladoceran populations

Acute zinc toxicity was assessed for 10 freshwater cladoceran species collected in six different ecosystems across Europe and for two standard laboratory-reared species (Daphnia magna and Ceriodaphnia dubia). The collected organisms belonged to five different genera: Daphnia (subgenus Daphnia and Ctenodaphnia), Ceriodaphnia, Simocephalus, Acroperus and Chydorus. The 48-h EC50 of the field-collected organisms tested in standard laboratory water ranged from 375+/-141 to 4314+/-1513 microg Znl(-1). The laboratory clone of D. magna was less sensitive than the majority of the field-collected species, while our laboratory Ceriodaphnia dubia was the second most sensitive. Considerable inter-species variation was found within the genus of Ceriodaphnia (factor 6) and within the genus Daphnia (factor 8). Among the different (sub)genera tested, Chydorus and Ctenodaphnia were significantly more tolerant than the others (up to a factor 3 difference). A significant positive relationship (r2=0.67, p<0.05) between the mean cladoceran 48-h EC50 and the ambient zinc concentration of the different aquatic systems was demonstrated, suggesting a role of acclimation and/or adaptation. No significant correlation between the acute zinc tolerance and the length of the organisms was found.     

Effect of cadmium on free amino acid, glutathione and ascorbic acid concentrations in two barley genotypes (Hordeum vulgare L.) differing in cadmium tolerance

Hydroponic experiment was carried out to study the effect of three Cd levels on glutathione (GSH), free amino acids (FAA), and ascorbic acid (ASA) concentration in the different tissues of 2 barley cultivars with different Cd tolerance. Cadmium concentration in both roots and shoots increased with external Cd level, while biomass and ASA concentration declined, and Wumaoliuling, a Cd-sensitive genotype was more affected than ZAU 3, a Cd-tolerant genotype. The effect of Cd on GSH concentration was dose- and time-dependent. In the 5 d exposure, root GSH concentration increased in 0.5 microM Cd treatment compared with control, but decreased significantly in 5 microM Cd treatment, irrespective of genotypes. However, in the 10 d exposure, GSH concentration in all plant tissues decreased with increasing Cd levels in the culture medium, and Wumaoliuling was much more affected than ZAU 3. Cadmium treatment greatly altered FAA concentration and composition in plants. The effect of Cd on glutathione (Glu) concentration in roots varied with genotypes. ZAU 3 showed a steady increase in root Glu concentration in both 0.5 and 5 microM Cd treatments, while Wumaoliuling was decreased by 38.0% in 5 microM Cd treatment, compared with the control. The results indicate that GSH and ASA are attributed to Cd tolerance in barley plants, and the relative less reduction in GSH concentration in ZAU 3 under Cd stress relative to the control may account for its higher Cd tolerance.     

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.     

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.     

Variation in the development of cadmium resistance in Daphnia magna straus; effect of temperature, nutrition, age and genotype

The extent to which responses to toxicants can be modified by the environment and genotype of Daphnia magna is important for interpreting the results from ecotoxicity tests. Variation in the development of cadmium resistance and its relevance for survivorship has been investigated in two genetically distinct clones of D. magna. Cadmium resistance was induced by pre-exposing daphnids to a sublethal concentration of a Cd/Zn mixture and thereafter cadmium tolerance was assessed in standard acute tests. The results showed that the ability to develop cadmium resistance is affected by temperature, but there is no consistent pattern for maternal nutrition. The age of the daphnids had no apparent effect on the development of cadmium resistance. Differences in the induction of cadmium resistance between clones were seen only under resource depression.     

The response of Populus spp. to cadmium stress: Chemical,morphological and proteomics study

Poplar (Populus) species are seen as candidates for removing heavy metal contamination from polluted soil. A bottom-up multidisciplinary approach was utilized to compare the performances of clones 58-861 and Poli (Populus nigra) and A4A, a Populus nigra × Populus deltoides hybrid to Cd toxicity. Qualitative and quantitative differences in their tolerance to Cd exposure and the uptake, accumulation and translocation of Cd were noted following the hydroponic exposure of rooted cuttings to 20 μM CdSO₄ for either 48 h or 14 d. Cadmium was less toxic for the hybrid clone A4A as compared to Poli and 58-861. Cd uptake and root to shoot translocation were determined by AAS, and its compartmentation was analyzed using SEM/EDX. A comparative proteomic approach was utilized to identify changes in proteins expression according to dose and time of exposure. Toxicity to Cd mainly influenced proteins related to general defense, stress response and carbohydrate metabolism.     

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.     

Growth and photosynthetic responses of the bloom-forming cyanobacterium Microcystis aeruginosa to elevated levels of cadmium

Effects of cadmium (Cd) on the growth and photosynthesis of the bloom-forming cyanobacterium Microcystis aeruginosa Kütz 854 were investigated. The growth was markedly inhibited when it was treated with 4 microM Cd. However, the biomass production was almost not influenced after a prolonged exposure at Cd concentrations < or = 2 microM. Chlorophyll content was more sensitive to Cd toxicity than phycobiliproteins at 0.5 microM Cd. However, the decrease of phycobiliproteins was larger than chlorophyll at the highest Cd concentration. A significant increase of F(v)/F(m) value was observed at Cd concentrations < or = 2 microM. On the other hand, when cells were treated with 4 microM Cd, F(v)/F(m) was significantly increased after 12 h of treatment but decreased after 48 h. The true photosynthesis was decreased with the increase of Cd concentration at 2 h. However, we noticed a recovery when the treatment was prolonged. After 48 h of exposure at the highest Cd concentration, photosynthetic oxygen evolution was markedly inhibited but dark respiration increased by 67%. Cellular Cd contents were augmented with the increase of Cd concentration. To our knowledge, we have demonstrated for the first time that the inhibitory site of Cd in M. aeruginosa is not located at the PSII or PSI level, but is probably situated on the ferredoxin/NADP(+)-oxidoreductase enzyme at the terminal of whole electron transport chain. We noticed also an increase of PSI activity, which is probably linked to the enhancement of cyclic electron transport around PSI. We can conclude that the increase of cyclic electron transport and dark respiration activities, and the decrease of phycobiliproteins might be adaptive mechanisms of M. aeruginosa 854 under high Cd conditions.     

Multi-generation studies with Chironomus riparius--effects of low tributyltin concentrations on life history parameters and genetic diversity

While toxicological data are available for numerous chemicals from standard tests, little is known on effects of pollutants over several generations or regarding chronic effects of chemicals on genetic diversity. Within the experiments, effects of the model pollutant tributyltin (TBT) were investigated over eleven generations at a sublethal TBT concentration of 4.46 μg as Sn kg⁻¹ sediment dw on life-cycle parameters and genetic variability of Chironomus riparius. Moreover, the adaptation potential towards TBT was determined. This experimental design enables the identification of TBT effects on life-cycle parameters and the determination of a potential extinction risk caused by chronic exposure. Furthermore, effects on the genetic structure can be determined, which are not predictable based solely on knowledge of the toxic mode of action of the chemical.

Genetic variety was determined via microsatellite analysis, measuring individual length differences of highly variable satellite DNA fragments. For the identification of changes in tolerances towards the stressor, acute and chronic toxicity experiments were conducted.

During the multi-generation study, significant effects on development and reproduction were determined. For some generations, the emergence was significantly (p < 0.05) delayed under TBT exposure. Reproduction seems to be a sensitive parameter as well, whereby females laid significantly larger egg masses (p < 0.05) in the latter generations. TBT did not affect the population growth rate nor the genetic variability, while clear deviations from the Hardy–Weinberg equilibrium appeared. The study also provides strong evidence for the acquirement of a higher tolerance towards the stressor in the TBT-exposed group.     

Effects of cadmium on the activities of photosystems of Chlorella pyrenoidosa and the protective role of cyclic electron flow

Cadmium (Cd) shows high toxicity to aquatic microalgae. Many studies showed that Cd inhibited activities of photosystem II (PSII) but the effects of heavy metals on photosystem I (PSI) and cyclic electron flow (CEF) were still controversial and unclear. The effects of CdCl₂ on the activities of PSI, PSII and CEF in Chlorella pyrenoidosa was measured simultaneously in the present study. In presence of 200 μM of Cd, ultrastructure of some cells was strongly modified. Cd exposure led to decrease of the activities of photosynthetic oxygen evolution and respiration. PSII was more sensitive to Cd treatment than PSI. Cd treatment showed significant inhibition on the photochemical quantum yield and electron transport rate of PSII. Cd increased the quantum yield of non-light-induced non-photochemical fluorescence quenching, indicating the damage of PSII. The activity of PSI showed tolerance to Cd treatment with concentration less than 100 μM in the experiment. Linear electron flow (LEF) made significant contribution to the photochemical quantum yield of PSI of the untreated cells, but decreased with increasing Cd concentration. The contribution of CEF to the yield of PSI increased with increasing Cd concentration. The activation of CEF after exposure to Cd played an essential role for the protection of PSI.     

Acute, chronic and sublethal effects of the herbicide propanil on Daphnia magna

Acute and chronic toxicity tests with propanil were conducted on Daphnia magna. The 24 and 48 h LC50 were 43.74 and 5.01 mg/l respectively. Chronic toxicity tests were carried out using sublethal propanil concentrations (0.07, 0.10, 0.21 and 0.55 mg/l) during 21 days. The effect of propanil on survival, reproduction and growth of D. magna organisms was monitored. The parameters used to evaluate herbicide effect on reproduction were: mean total young ones per female, mean brood size, time to first reproduction, mean number broods per female and intrinsic rate of natural increase (r). Survival and growth (body length) were also determined after 21 days of exposure to the herbicide. Reproduction was significantly reduced when propanil concentration increased in the medium. The intrinsic rate of natural increase (r) decreased with increasing concentrations of propanil especially in those animals exposed to 0.55 mg/l. However, growth as well as survival of the exposed organisms only decreased in daphnids exposed to the highest propanil concentration tested. The maximum acceptable toxicant concentration (MATC) was calculated for D. magna exposed to the herbicide using as parameter of evaluation the intrinsic rate of natural increase (r). The interpolation of these results gave MATC values of 0.08 mg/l herbicide. We have derived the EC50 values for some selected parameters on D. magna exposed to propanil. EC50 values indicated that reproductive parameters were very sensitive of the effect of propanil on daphnids. Finally, the daphnids were exposed to the same sublethal herbicide concentrations as in the chronic study and the effect of the toxicant on filtration and ingestion rates was determined. Feeding rates of D. magna declined with increasing propanil concentrations. The effective propanil concentrations at which feeding rates were reduced to 50% of that in controls (EC50) were also calculated.     

Ecotoxicological evaluation of sodium fluoroacetate on aquatic organisms and investigation of the effects on two fish cell lines

Sodium monofluoroacetate (compound 1080) is one of the most potent pesticides. It is also a metabolite of many other fluorinated compounds, including anticancer agents, narcotic analgesics, pesticides or industrial chemicals. Other sources of water contamination are the atmospheric degradation of hydrofluorocarbons and hydrochlorofluorocarbons. However, there is little information available about the adverse effects of sodium fluoroacetate in aquatic organisms. Firstly, the bacterium Vibrio fischeri (decomposer), the alga Chlorella vulgaris (1st producer) and the cladoceran Daphnia magna (1st consumer) were used for the ecotoxicological evaluation of SMFA. The most sensitive models were C. vulgaris and D. magna, with a NOAEL of 0.1 and an EC50 of 0.5 mM at 72 h, respectively. According to the results after the acute exposure and due to its high biodegradation rate and low bioaccumulation potential, sodium fluoroacetate is most unlikely to produce deleterious effects to aquatic organisms. Secondly, two fish cell lines were employed to investigate the effects and mechanisms of toxicity in tissues from 2nd consumers. The hepatoma fish cell line PLHC-1 was more sensitive to SMFA than the fibroblast-like fish cell line RTG-2, being the uptake of neutral red the most sensitive bioindicator. Lysosomal function, succinate dehydrogenase and acetylcholinesterase activities were inhibited, glucose-6-phosphate dehydrogenase activity was particularly stimulated, and metallothionein and ethoxyresorufin-O-deethylase levels were not modified. Intense hydropic degeneration, macrovesicular steatosis and death mainly by necrosis but also by apoptosis were observed. Moreover, sulphydryl groups and oxidative stress could be involved in PLHC-1 cell death induced by SMFA more than changes in calcium homeostasis.     

Quantification of differentially expressed genes in Daphnia magna exposed to rubber wastewater

In this study, differentially expressed genes (DEGs) were investigated in Daphnia magna exposed to rubber wastewater using an annealing control primer (ACP)-based polymerase chain reaction (PCR) and real-time PCR. Among three identified DEGs, two genes (DEG1 and DEG2) were up-regulated, and DEG1 expression was well-correlated to a logarithm of rubber wastewater concentration (r(2)=0.971, p<0.0001). In addition, DEG1 expression in D. magna exposed to rubber wastewater was strongly correlated with that of D. magna exposed to Zn (r(2)=0.9513, p<0.05), suggesting that the induction of DEG1 was caused by Zn, which is the dominant toxicant in rubber wastewater. In addition, DEG1 expression was more sensitive to toxicants than immobility, which is the conventional endpoint in toxicity tests using D. magna. The lowest observed effect concentrations (LOEC) determined using immobility tests were 2.5% for rubber wastewater and 1.6mgl(-1) for Zn. In contrast, a significant increase in DEG1 expression was observed at exposure concentrations of as low as 0.6% rubber wastewater and 0.2mgl(-1) Zn. These results indicate that DEG1 is a sensitive and quantitative biomarker of water and wastewater containing Zn.