Cloning metallothionein gene in Zacco platypus and its potential as an exposure biomarker against cadmium.

Zacco platypus, pale chub, is an indigenous freshwater fish of East Asia including Korea and has many useful characteristics as indicator species for water pollution. While utility of Z. platypus as an experimental species has been recognized, genetic-level information is very limited and warrants extensive research. Metallothionein (MT) is widely used and well-known biomarker for heavy metal exposure in many experimental species. In the present study, we cloned MT in Z. platypus and evaluated its utility as a biomarker for metal exposure. For this purpose, we sequenced complete complementary DNA (cDNA) of MT in Z. platypus and carried out phylogenetic analysis with its sequences. The transcription-level responses of MT gene following the exposure to CdCl2 were also assessed to validate the utility of this gene as an exposure biomarker. Analysis of cDNA sequence of MT gene demonstrated high conformity with those of other fish. MT messenger RNA (mRNA) expression and enzymatic MT content significantly increased following CdCl2 exposure in a concentration-dependent manner. The level of CdCl2 that resulted in significant MT changes in Z. platypus was within the range that was reported from other fish. The MT gene of Z. platypus sequenced in the present study can be used as a useful biomarker for heavy metal exposure in the aquatic environment of Korea and other countries where this freshwater fish species represents the ecosystem.

Non-monotonic concentration-response relationship of TiO(2) nanoparticles in freshwater cladocerans under environmentally relevant UV-A light.

The effects of UV-A on the toxicity of TiO2 nano-particles (NPs) were evaluated using Moina macrocopa and Daphnia magna under environmentally relevant level of UV-A. The waterfleas were exposed to TiO2 NPs with different sizes of ~298nm, ~132nm, or ~72nm for up to 48h, with or without UV-A light. Whole body reactive oxygen species and transcription of antioxidant enzyme genes were measured, as well as the survival of the waterflea. In the presence of UV-A, the survival rates of M. macrocopa significantly decreased in concentration dependent way until ~1mg/L TiO2 NPs, but the survivals were reversed at greater concentrations. This peculiar non-monotonic trend of concentration-response relationship might be explained by changes of particle size under different light conditions. TiO2 NPs within a certain size range could be trapped in the filter apparatus and exert toxicity, and the NPs of greater size were subject to either precipitation or ingestion leading to no or little toxicity. Observed TiO2 toxicity was associated with oxidative stress in the filter apparatus. The results of this study showed that the size change due to UV-A irradiation should be considered in evaluation of ecological risks of TiO2 NP.

Effects of ionization on the toxicity of silver nanoparticles to Japanese medaka (Oryzias latipes) embryos.

Increase in the use of manufactured nanomaterials (NMs) has led to concerns about the environmental impacts. Especially, hazard of metal-based NMs is more severe due to ions released from surface by water quality parameters and physicochemical properties after entering into the water environment. However, little is known about the effects of ionization on the toxicity of metal-based NMs in the water environment. To address this question, we prepared the suspensions of silver nanoparticles (AgNP) at 25 µg L(-1) containing different concentrations of Ag(+) (5, 10, 20, 45, and 75% Ag(+) to total Ag), and evaluated their toxicity to Japanese medaka (Oryzias latipes) embryos. Higher Ag(+) ratios in the AgNP suspension, suggesting the lower number of particles, led to the higher adverse effects on embryos and sac-fries. In addition, histopathology analysis revealed that AgNPs penetrated through chorion of eggs and skin membrane, and were distributed into the tissues. The results imply that the ionization could decrease the toxicity of metal-based NMs in the water environment.

Tissue-specific antioxidant responses in pale chub (Zacco platypus) exposed to copper and benzo[a]pyrene.

In this study, antioxidant responses including lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST), were evaluated in the liver, gill and muscle tissues of pale chub (Zacco platypus) exposed to copper (Cu) and benzo[a]pyrene (BaP). Cu exposure induced significant antioxidant responses in Z. platypus, particularly in the liver, whereas BaP exposure had a negligible effect. Following Cu exposure, both SOD and CAT activity increased in a concentration-dependent manner, showing significant correlations with malondialdehyde (MDA) levels as a measure of LPO (r = 0.646 and 0.663, respectively). SOD, CAT and GST mRNA levels were also enhanced following Cu exposure, except at 20 µg L(-1), although significant correlations with antioxidant enzyme activities were not found. The results of this study suggest that combined information on SOD and CAT activities together with LPO levels in the liver could be a useful indicator for assessing oxidative stress in freshwater fish.

Soluble c-Met protein as a susceptible biomarker for gastric cancer risk: A nested case-control study within the Korean Multicenter Cancer Cohort.

This study was conducted to evaluate the relevance of the soluble form of c-Met protein, a truncated form of the c-Met membrane receptor involved in the CagA pathway, as a potential biomarker for gastric cancer. Among 290 gastric cancer case-control sets selected from the Korean Multicenter Cancer Cohort, the plasma concentrations of soluble c-Met protein were measured with enzyme-linked immunosorbent assays. Using analysis of variance and covariance models with age, sex, smoking, Helicobacter pylori infection, and CagA seropositivity, the mean concentrations of soluble c-Met protein between cases and controls were compared. To evaluate the association between gastric cancer and a c-Met protein level, odds ratios and 95% confidence intervals were estimated using conditional logistic regression models. Interactions between CagA-related genes and the soluble c-Met protein concentration were also investigated. The overall median plasma concentration of soluble c-Met among cases was significantly lower than those of controls (1.390 vs. 1.610 ng/mL, p < 0.0001). Closer to the onset of gastric cancer, the soluble c-Met protein level decreased linearly in a time-dependent manner (p for trend = 0.0002). The combined effects between the CagA-related genes and the soluble c-Met protein concentration significantly intensified risks for gastric cancer. Restricted analyses including cases that had been diagnosed within 1 year after entering the cohort had a fair degree of ability (area under the receiver operating characteristic curve of 0.73-0.77) to discriminate gastric cancer cases from normal controls. Our findings demonstrate the potential of the soluble form of c-Met protein as a novel biomarker for gastric cancer. The beneficial effects of a high soluble c-Met concentration in human plasma are strongly supported.

Potential application of urease and nitrification inhibitors to mitigate emissions from the livestock sector: a review.

Human activities have caused an increase in greenhouse gas emissions, resulting in climate change that affects many factors of human life including its effect on water and food quality in certain areas with implications for human health. CH4 and N2O are known as potent non-CO2 GHGs. The livestock industry contributes to direct emissions of CH4 (38.24%) and N2O (6.70%) through enteric fermentation and manure treatment, as well as indirect N2O emissions via NH3 volatilization. NH3 is also a secondary precursor of particulate matter. Several approaches have been proposed to address this issue, including dietary management, manure treatment, and the possibility of inhibitor usage. Inhibitors, including urease and nitrification inhibitors, are widely used in agricultural fields. The use of urease and nitrification inhibitors is known to be effective in reducing nitrogen loss from agricultural soil in the form of NH3 and N2O and can further reduce CH4 as a side effect. However, the effectiveness of inhibitors in livestock manure systems has not yet been explored. This review discusses the potential of inhibitor usage, specifically of N-(n-butyl) thiophosphoric triamide, dicyandiamide, and 3,4-dimethylpyrazole phosphate, to reduce emissions from livestock manure. This review focuses on the application of inhibitors to manure, as well as the association of these inhibitors with health, toxicity, and economic benefits.

Determination of mRNA expression of DMRT93B, vitellogenin, and cuticle 12 in Daphnia magna and their biomarker potential for endocrine disruption.

We explored the use of molecular genetic biomarkers for endocrine disruption in Daphnia magna after the exposure to fenoxycarb (FOC), a model juvenile hormone analog. For this purpose, the mRNA expression patterns of DMRT93B (DMRT, sex determination), cuticle 12 (CUT, molting), and vitellogenin (VTG, embryo development) were determined in D. magna. Furthermore, these results were compared with developmental abnormality and reproduction performance. The fold changes of CUT and VTG mRNA expression showed significant dose-response relationship with FOC exposure. Relative mRNA expressions of DMRT and CUT showed notable changes at as low as 1 ng/l FOC. After chronic exposure FOC significantly delayed the first day of reproduction and decreased the number of young and growth rate even at 10 ng/l FOC. A concentration-dependant trend in reproduction effect was also observed. Developmental abnormality such as poorly developed second antennae and curved or unextended shell spines were observed. These results suggest that the three mRNAs, i.e., DMRT, CUT, and VTG can be used as biomarkers of endocrine disrupting effects in D. magna.

Implication of global environmental changes on chemical toxicity-effect of water temperature, pH, and ultraviolet B irradiation on acute toxicity of several pharmaceuticals in Daphnia magna.

Global environmental change poses emerging environmental health challenges throughout the world. One of such threats could be found in chemical safety in aquatic ecosystem. In the present study, we evaluated the effect of several environmental factors, such as water pH, temperature and ultraviolet light on the toxicity of pharmaceutical compounds in water, using freshwater invertebrate Daphnia magna. Seven pharmaceuticals including ibuprofen, acetaminophen, lincomycin, ciprofloxacin, enrofloxacin, chlortetracycline and sulfathiazole were chosen as test compounds based on their frequent detection in water. The experimental conditions of environmental parameters were selected within the ranges that could be encountered in temperate environment, i.e., water temperature (15, 21, and 25 degrees C), pH (7.4, 8.3, and 9.2), and UV-B light intensity (continuous irradiation of 15.0 microW/cm(2)). For acetaminophen, enrofloxacin and sulfathiazole, decrease in water pH generally led to increase of acute lethal toxicity, which could be explained by the unionized fraction of pharmaceuticals. Increase of water temperature enhanced the acute toxicity of the acetaminophen, enrofloxacin and chlortetracycline, potentially due to alteration in toxicokinetics of chemicals as well as impact on physiological mechanisms of the test organism. The presence of UV-B light significantly increased the toxicity of sulfathiazole, which could be explained by photo-modification of this chemical that lead to oxidative stress. Under the UV light, however, acute toxicity of enrofloxacin decreased, which might be due to photo-degradation. Since changing environmental conditions could affect exposure and concentration-response profile of environmental contaminants, such conditions should be identified and evaluated in order to better manage ecosystem health under changing global environment.

Acute toxicity of two CdSe/ZnSe quantum dots with different surface coating in Daphnia magna under various light conditions.

With an increasing use of quantum dots (QDs) in many applications, their potential hazard is of growing concern. However, little is known about their ecotoxicity, especially in vivo. In the present study, we employed freshwater macroinvertebrate, Daphnia magna, to evaluate toxicity characteristics of cadmium selenide/zinc selenide (CdSe/ZnSe) in relation to surface coatings, e.g., mercaptopropionic acid QD ((MPA)QD), and gum arabic/tri-n-octylphosphine oxide QD ((GA/TOPO)QD), and light conditions, i.e., dark, fluorescent light, environmental level of ultraviolet (UV) light, and sunlight. The results of the present study showed that D. magna was more susceptible to (GA/TOPO)QD exposure compared to (MPA)QD. The surface coating of QD appeared to determine the stability of QDs and hence the toxicity, potentially by size change of or the release of toxic components from QDs. However, (GA/TOPO)QD was still less toxic than the equivalent level of CdCl2. The toxicity of all the tested compounds increased by changing the light condition from dark to white fluorescence to UV-B light, and to natural sunlight. The effect of light condition on QDs toxicity could also be explained by photostability of the QDs, which would affect size of the particle, release of toxic component ions, and generation of reactive oxygen species. Considering increasing use of QDs in various applications, their environmental fates and corresponding toxic potentials deserve further investigation.

Application of a microbial toxicity assay for monitoring treatment effectiveness of pentachlorophenol in water using UV photolysis and TiO2 photocatalysis.

Conventional approaches for monitoring the effectiveness of wastewater treatment processes include evaluating the degradation of the target compound and/or generation of its nontoxic byproducts. These approaches are, however, limited because routine chemical analyses alone are neither able to fully address potential hazard to biological receptors nor characterize potential synergistic interactions. This study was carried out to investigate the degradation effectiveness of pentachlorophenol (PCP) by treatment with UV-A, UV-B photolysis, sunlight, TiO(2) photocatalysis, and/or their combinations. Chemical analyses of the parent compound and its selected byproducts, as well as acute toxicity assessment using the luminescent bacteria Vibrio fischeri (Microtox), were conducted during and after the various photolytic and photocatalytic treatments. In general, the toxicity reduction pattern observed after treatment corresponded well with the chemical degradation data. However, it should be noted that there were occasions that acute microbial toxicity was observed even from the treated water samples, some of which showed complete removal of the parent compound. This post-treatment toxicity might be due to toxic PCP byproducts, which may include polychlorinated dibenzodioxins/furans, tetrachloro-p-benzoquinone, and other intermediates. The TiO(2) photocatalysis with UV-B photolysis was the most effective method to remove both PCP and its toxic derivatives in the water. The Microtox assay is an easy to use and promising approach for evaluating the effectiveness of wastewater treatment processes.

Degradation mechanism and the toxicity assessment in TiO2 photocatalysis and photolysis of parathion.

The photocatalytic degradation of methyl parathion was carried out using a circulating TiO2/UV reactor. The experimental results showed that parathion was more effectively degraded in the photocatalytic condition than the photolysis and TiO2-only condition. With photocatalysis, 10mg/l parathion was completely degraded within 60 min with a TOC decrease exceeding 90% after 150 min. The main ionic byproducts during photocatalysis were measured. The nitrogen from parathion was recovered mainly as NO3-, NO2- and NH4+, 80% of the sulfur as SO4(2-), and less than 5% of the phosphorus as PO4(3-). The organic intermediates 4-nitrophenol and paraoxon were also identified, and these were further degraded. Two different bioassays (Vibrio fischeri and Daphnia magna) were used to test the acute toxicity of solutions treated by photocatalysis and photolysis. A Microtox test using V. fischeri showed that the toxicity, expressed as the relative toxicity (%), was reduced almost completely after 90 min under photocatalysis, whereas only an 83% reduction was achieved with photolysis alone. Another toxicity test using D. magna also showed that the relative toxicity disappeared after 90 min under photocatalysis, whereas there was a 65% reduction in relative toxicity with photolysis alone. The pattern of toxicity reduction parallels the decrease in parathion and TOC concentrations.

Combined effects of the cyanobacterial toxin microcystin-LR and environmental factors on life-history traits of indigenous cladoceran Moina macrocopa.

Environmental factors are important in that they might interact with toxicants and could affect the concentration-response relationship of the toxicants in the water environment. Microcystins (MCs) produced by algal blooming are natural toxins that exert severe impacts on aquatic organisms. Despite the possibility of interaction effects between environmental factors and MCs, very few studies have been carried out to date. In the present study, the authors evaluated the effect of 3 environmental factors on chronic toxicity of MC-leucine-arginine (MC-LR; 0 µg/L, 0.8 µg/L, 4 µg/L, 20 µg/L, 100 µg/L, 500 µg/L) to the freshwater invertebrate Moina macrocopa. Three environmental factors were determined to reflect the reasonable worst conditions of the water body in South Korea: high water temperature (25 °C), the highest concentration of nutrients (ammonia-nitrogen [NH3 -N] = 3.8 mg/L; nitrate-nitrogen [NO3 -N] = 8.5 mg/L) during the occurrence of algal bloom, and 2 pH conditions of 7.0 and 9.0, which satisfy the test acceptance criteria for the Daphnia test. Among the various environmental factors being tested, high water temperature and NO3 -N elevated chronic toxicity of MC-LR, whereas NH3 -N reduced toxicity. Water pH did not influence chronic toxicity of MC-LR to M. macrocopa. This observation suggests that those environmental factors are responsible for changing the trend of MC-LR toxicity.

Stepwise embryonic toxicity of silver nanoparticles on Oryzias latipes.

The developmental toxicity of silver nanoparticles (AgNPs) was investigated following exposure of Oryzias latipes (medaka) embryos to 0.1-1 mg/L of homogeneously dispersed AgNPs for 14 days. During this period, developmental endpoints, including lethality, heart rate, and hatching rate, were evaluated by microscopy for different stages of medaka embryonic development. To compare toxic sensitivity, acute adult toxicity was assessed. There was no difference in acute lethal toxicity between embryo and adult medaka. Interestingly, we found that the increase in stepwise toxicity was dependent on the developmental stage of the embryo. Lethal embryonic toxicity increased from exposure days 1 to 3 and exposure days 5 to 8, whereas there was no change from exposure days 3 to 5. In addition, 7 d exposure to 0.8 mg/L AgNPs resulted in significant heart beat retardation in medaka embryos. AgNPs also caused a dose-dependent decrease in the hatching rate and body length of larvae. These results indicate that AgNP exposure causes severe developmental toxicity to medaka embryos and that toxicity levels are enhanced at certain developmental stages, which should be taken into consideration in assessments of metallic NPs toxicity to embryos.

Assessment of exposure to heavy metals and health risks among residents near abandoned metal mines in Goseong, Korea.

Metal contamination from mining activity is of great concern because of potential health risks to the local inhabitants. In the present study, we investigated the levels of Cd, Cu, As, Pb, and Zn in environmental samples and foodstuffs grown in the vicinity of the mines in Goseong, Korea, and evaluated potential health risks among local residents. Soils near the mines exceeded the soil quality standard values of Cu, As, and Zn contamination. The concentrations of Cd, Cu, Pb, and Zn in crop samples collected from the study area were significantly higher than those of the reference area. Some rice samples collected from the study area exceeded the maximum permissible level of 0.2 mg Cd/kg. The intake of rice was identified as a major contributor (≥75%) to the estimated daily intake among the residents. The average estimated daily intakes of metals were, however, below the provisional tolerable daily intake.

Perfluorooctane sulfonic acid exposure increases cadmium toxicity in early life stage of zebrafish, Danio rerio.

Exposure to perfluorooctane sulfonic acid (PFOS) is known to induce thyroid-related adverse effects in aquatic organisms. Because an antioxidant defense mechanism is one of the key functions of the thyroid gland, we examined whether preexposure to PFOS could disrupt thyroid function and enhance cadmium (Cd)-induced oxidative stress in fish. Zebrafish embryos were exposed to control or 0.5 mg/L PFOS for 7 d after fertilization and subsequently exposed to 0.038 mg/L of Cd(2+) or a mixture of the PFOS and Cd for an additional 3 d until 10 d postfertilization (dpf). Survival rates, body length, messenger RNA (mRNA) expressions related to thyroid function and oxidative stress, the levels of thyroid hormones, and malondialdehyde and antioxidant enzyme activities were measured. Significant down-regulation of mRNAs related to thyroid function (thyroid hormone receptor-alpha [THRα], thyroid hormone receptor-beta [THRß], hematopoietically expressed homeobox [hhex], and paired box gene 8 [pax8]) and decrease of throxine (T4) levels were observed in the PFOS preexposure group, suggesting that PFOS preexposure would influence the performance of thyroid gland in the later stages of life. Certain genes relative to oxidative stress, such as superoxide dismutase 1 (sod1) and heat shock protein 70 (hsp70), in the PFOS preexposure group were significantly up-regulated when the larvae were subsequently exposed to Cd or to the mixture of PFOS and Cd. Glutathione S-transferase activity and malondialdehyde levels of the PFOS-preexposed group were increased significantly by Cd exposure. Significant decrease of the survival rates and body length of fish were observed at 10 dpf among the larvae that were previously exposed to PFOS. These results suggest that preexposure to PFOS could affect antioxidant defense mechanisms and potentially increase the toxicity of Cd on mRNA expression and enzyme activity level responses, as well as on survival or growth of individuals.

Phototoxicity of CdSe/ZnSe quantum dots with surface coatings of 3-mercaptopropionic acid or tri-n-octylphosphine oxide/gum arabic in Daphnia magna under environmentally relevant UV-B light.

The potential ecotoxicological consequences about semiconductor crystal nanoparticles (NPs) are a growing concern. However, our understanding of the mechanism of toxicity in NPs is very limited, especially under varying environmental conditions such as ultraviolet (UV) light. We performed an in vivo study employing Daphnia magna to evaluate the mechanism involved in toxicity of cadmium selenide/zinc selenide quantum dots (QDs) with two different organic coatings under an environmental level of UV-B light. We used QDs with mercaptopropionic acid (MPA) and tri-n-octylphosphine oxide/gum arabic (GA) and measured their toxicities under an environmental level of UV-B light. Whole-body reactive oxygen species (ROS) generation and mRNA expression level biomarkers, as well as acute toxicity, were measured in D. magna. With UV-B light, both cadmium (Cd) and GA-QD became more toxic in daphnids. The levels of small Cd molecules (<10kDa cutoff) increased for GA-QD under UV-B; however, the observed acute lethal toxicity could not be explained by the measured Cd level. Under UV-B light, both Cd and GA-QD generated more ROS. In addition, the expression pattern of mRNAs specific to Cd exposure was not observed from GA-QD with or without UV-B light. These observations suggest that the phototoxicity of QDs may be explained not only by Cd release from the QD core but also by stability of surface coating characteristics and other potential causes such as ROS generation.

Molecular cloning of Daphnia magna catalase and its biomarker potential against oxidative stresses.

Catalase (EC 1.11.1.6) is an important antioxidant enzyme that protects aerobic organisms against oxidative damage by degrading hydrogen peroxide to oxygen and water. Catalase mRNAs have been cloned from many species and employed as useful biomarkers of oxidative stress. In the present study, we cloned the cDNA from the catalase gene in Daphnia magna, analyzed its catalytic properties, and investigated mRNA expression patterns after the exposure to known oxidative stressors. The catalase proximal heme-ligand signature sequence, FDRERISERVVHAKGSGA, and the proximal active site signature, RLFSYTDTH, are highly conserved. The variation of catalase mRNA expression in D. magna was quantified by real-time PCR, and the results indicated that catalase expression was up-regulated after exposure to UV-B light or cadmium (Cd). The activity of catalase enzyme also showed a similar increasing pattern when exposed to these model stressors. The full-length catalase cDNA of D. magna was cloned using mixed primers by the method of 3' and 5' rapid amplification of cDNA ends PCR. The cDNA sequence consists of 1515 nucleotides, encoding 504 amino acids. Sequence comparison showed that the deduced amino acid sequence of D. magna shared 73%, 72%, 71% and 70% identity with that of Chlamys farreri, Fenneropenaeus chinensis, Litopenaeus vannamei and Anopheles gambiae, respectively. This study shows that the catalase mRNA from D. magna could be successfully employed as a biomarker of oxidative stress, which is a common mode of toxicity for many water contaminants.

Endocrine disruption and consequences of chronic exposure to ibuprofen in Japanese medaka (Oryzias latipes) and freshwater cladocerans Daphnia magna and Moina macrocopa.

Despite frequent detection of ibuprofen in aquatic environments, the hazards associated with long-term exposure to ibuprofen have seldom been investigated. Ibuprofen is suspected of influencing sex steroid hormones through steroidogenic pathways in both vertebrates and invertebrates. In this study, the effect of ibuprofen on sex hormone balance and the associated mechanisms was investigated in vitro by use of H295R cells. We also conducted chronic toxicity tests using freshwater fish, Oryzias latipes, and two freshwater cladocerans, Daphnia magna and Moina macrocopa, for up to 144 and 21d of exposure, respectively. Ibuprofen exposure increased 17beta-estradiol (E2) production and aromatase activity in H295R cells. Testosterone (T) production decreased in a dose-dependent manner. For D. magna, the 48 h immobilization EC50 was 51.4 mg/L and the 21 d reproduction NOEC was <1.23 mg/L; for M. macrocopa, the 48 h immobilization EC50 was 72.6 mg/L and the 7d reproduction NOEC was 25mg/L. For O. latipes, 120 d survival NOEC was 0.0001 mg/L. In addition, ibuprofen affected several endpoints related to reproduction of the fish, including induction of vitellogenin in male fish, fewer broods per pair, and more eggs per brood. Parental exposure to as low as 0.0001 mg/L ibuprofen delayed hatching of eggs even when they were transferred to and cultured in clean water. Delayed hatching is environmentally relevant because this may increase the risk of being predated. For O. latipes, the acute-to-chronic ratio of ibuprofen was estimated to be greater than 1000. Overall, relatively high acute-to-chronic ratio and observation of reproduction damage in medaka fish at environmentally relevant ranges of ibuprofen warrant the need for further studies to elucidate potential ecological consequences of ibuprofen contamination in the aquatic environment.

Phototoxicity and oxidative stress responses in Daphnia magna under exposure to sulfathiazole and environmental level ultraviolet B irradiation.

Sulfonamide antibiotics frequently occur in aquatic environments. In this study, phototoxicity of sulfathiazole (STZ) and its mechanism of action were investigated using Daphnia magna. We evaluated the changes of molecular level stress responses by assessing gene expression, enzyme induction and lipid peroxidation, and the related organism-level effects in D. magna. In the presence of ultraviolet B (UV-B) light (continuous irradiation with 13.8+/-1.0microWcm(-2)d(-1)), STZ (at the nominal concentration of 94.9mg/L) caused a significant increase in reactive oxygen species (ROS) generation and lipid peroxidation. Catalase (CAT) and glutathione S-transferase (GST) showed concentration-dependent increases caused by the exposure. Exposure to STZ and UV-B light caused apparent up-regulation of alpha-esterase, hemoglobin, and vitellogenin mRNA. The survival of daphnids was significantly affected by the co-exposure to STZ and UV-B. The biochemical and molecular level observations in combination with organism-level effects suggest that the phototoxicity of STZ was mediated in part by ROS generated by oxidative stress in D. magna.

Acclimation to ultraviolet irradiation affects UV-B sensitivity of Daphnia magna to several environmental toxicants.

Phototoxicity of several environmental contaminants by UV light has been reported in many studies. Nevertheless, field observations suggest the presence of certain defense mechanisms that would protect aquatic organisms against phototoxic damages. The current study was conducted to understand the responses of aquatic receptors to phototoxic chemicals in a natural environment where low dose UV light is present and long-term acclimation to UV might have been taken place. For this purpose, the water flea Daphnia magna was acclimated to a non-lethal, environmentally relevant level of UV-B light for >20 successive generations. The differences in toxicity response were evaluated between the UV-B acclimated and the non-acclimated daphnids when they were exposed to phototoxic compounds such as polynuclear aromatic hydrocarbons (fluoranthene and pyrene), a pharmaceutical (sulfathiazole), or metals (Cd and Cu) under UV-B light. Following the UV-B acclimation, toxicity of metals under UV-B light significantly decreased (P<0.1) suggesting the defense/repair system which might be developed through acclimation. For PAHs and sulfathiazole, however the acclimation rendered organisms more susceptible (P<0.05). The metabolic cost incurred during the acclimation to UV-B stress may in part explain the organisms' reduced capacity to deal with other stressors. Addition of vitamin C significantly increased the resistance of UV-B acclimated individuals against Cu, while no change was observed for the other chemicals, suggesting that the mode of Cu phototoxicity is different from those of the other phototoxicants under UV-B light. Two-dimensional gel electrophoresis analyses showed that long-term acclimation to UV-B lead to notable changes in protein expression, which may be further evaluated to explain varying susceptibilities of the acclimated daphnids to different phototoxicants.