Hepatic antioxidant status and hematological parameters in rainbow trout,Oncorhynchus mykiss,after chronic exposure to carbamazepine

Recently, residual pharmaceuticals are generally recognized as relevant sources of aquatic environmental pollutants. However, the toxicological effects of these contaminants have not been adequately researched. In this study, the chronic toxic effect of carbamazepine (CBZ), an anticonvulsant drug commonly present in surface and ground water, on hepatic antioxidant status and hematological parameters of rainbow trout were investigated. Fish were exposed at sublethal concentrations of CBZ (1.0 μg/l, 0.2 mg/l and 2.0 mg/l) for 7, 21 and 42 days. Compared to the control group, fish exposed at higher concentration (0.2 mg/l or 2.0 mg/l) of CBZ showed significantly higher levels of hemoglobin, ammonia and glucose, and significantly higher plasma enzymes activities. During the exposure duration, erythrocyte count, hematocrit, mean erythrocyte hemoglobin, mean erythrocyte volume, mean color concentration and total protein content in all groups were not significantly different. At the highest test concentration (2.0 mg/l) of CBZ, oxidative stress was apparent as reflected by the significant higher lipid peroxidation and protein carbonyl levels in liver after 42 days exposure, associated with an inability to induce antioxidant enzymes activities including superoxide dismutase, glutathione peroxidase and glutathione reductase. After 42 days exposure, reduced glutathione level was significantly decreased in the fish exposed at 0.2 mg/l CBZ, compared with the control. In short, CBZ-induced physiological and biochemical responses in fish were reflected in the oxidant stress indices and hematological parameters. These results suggest that hepatic antioxidant responses and hematological parameter could be used as potential biomarkers for monitoring residual pharmaceuticals present in aquatic environment.     

Effects of manufactured nanomaterials on fishes: a target organ and body systems physiology approach

Manufactured nanomaterials (NM) are already used in consumer products and exposure modelling predicts releases of ng to low μg l(-1) levels of NMs into surface waters. The exposure of aquatic ecosystems, and therefore fishes, to manufactured NMs is inevitable. This review uses a physiological approach to describe the known effects of NMs on the body systems of fishes and to identify the internal target organs, as well as outline aspects of colloid chemistry relevant to fish biology. The acute toxicity data, suggest that the lethal concentration for many NMs is in the mg l(-1) range, and a number of sublethal effects have been reported at concentrations from c. 100 μg to 1 mg l(-1). Exposure to NMs in the water column can cause respiratory toxicity involving altered ventilation, mucus secretion and gill pathology. This may not lead, however, to overt haematological disturbances in the short term. The internal target organs include the liver, spleen and haematopoietic system, kidney, gut and brain; with toxic effects involving oxidative stress, ionoregulatory disturbances and organ pathologies. Some pathology appears to be novel for NMs, such as vascular injury in the brain of rainbow trout Oncorhynchus mykiss with carbon nanotubes. A lack of analytical methods, however, has prevented the reporting of NM concentrations in fish tissues, and the precise uptake mechanisms across the gill or gut are yet to be elucidated. The few dietary exposure studies conducted show no effects on growth or food intake at 10-100 mg kg(-1) inclusions of NMs in the diet of O. mykiss, but there are biochemical disturbances. Early life stages are sensitive to NMs with reports of lethal toxicity and developmental defects. There are many data gaps, however, including how water quality alters physiological responses, effects on immunity and chronic exposure data at environmentally relevant concentrations. Overall, the data so far suggest that the manufactured NMs are not as toxic as some traditional chemicals (e.g. some dissolved metals) and the innovative, responsible, development of nanotechnology should continue, with potential benefits for aquaculture, fisheries and fish health diagnostics.     

Using Quantitative Structure–Activity Relationships to Support the Assessment of the Environmental Fate and Aquatic Toxicity of a Series of Methacrylic Acid Esters

Methacrylic acid esters or methacrylates (with C1 to 8 alkyl side-chains) are a group of compounds used in the manufacture of resins and plastics. Releases to the atmosphere, surface waters, or soil can come about during manufacturing, use, transport, and storage activities. Given the potential for environmental exposures, an examination into factors that control the distribution, fate, and toxicity of methacrylates is warranted. Methacrylates are relatively volatile compounds that are easily and rapidly degraded in the atmosphere and in oxic surface waters and sediments, and so are not considered persistent. Aquatic organisms are also able to metabolize methacrylates as demonstrated by estimated bioconcentration factors of < 45 for the acid and the C1-4 esters, and a measured bioconcentration factor (BCF) for 2-EHMA. Thus, the methacrylates are not considered bioaccumulative. Acute and chronic aquatic toxicity data for the methacrylates were assembled. Quantitative structure–activity relationships (QSARs) were developed with the acute and chronic fish, invertebrate, and algal datasets to support the toxicity assessment of the methacrylates (with log octanol-water partition coefficients used as surrogates for structures). The data show the methacrylates to be of low to moderate toxicity with all acute LC/EC50 values and chronic no observed effect concentrations ranging from about 2 to 170 mg/l for the acid and C1-4 esters, and 0.1 mg/l up to the aqueous solubility limit (about 2 mg/l) for the C8 ester. The measured toxicity data and the data estimated from the QSARs were used to develop predicted no effect concentrations in water and sediment for methacrylic acid and the C1-C8 esters.     

基于在线生物监测系统的氨氮胁迫下斑马鱼行为响应

研究基于四级阻抗技术的在线生物监测系统, 参照国家地表水环境质量标准(GB3838-2002)中对 Ⅰ 类和 Ⅳ 类地表水氨氮质量标准的限值分类, 探究 0.15和1.5 mg/L氨氮胁迫下 15d 内斑马鱼(Danio rerio)行为响应, 为环境胁迫下斑马鱼的行为响应提供数据支撑。实验采用斑马鱼游动强度和速率综合行为强度来反映其行为变化, 对实验数据进行 SPSS 差异性分析、自相关分析(Autocorrelation)和自组织神经网络(Self-organization mapping net, SOM)分析。研究结果: (1)在对照组中, 行为强度数据平均值为0.69±0.13, 光周期行为强度数据(0.73±0.13)高于暗周期(0.66±0.14; P<0.01), 且光暗周期差异值为10.61%; 在氨氮胁迫组(NH₃-N, 0.15和 1.5 mg/L)实验中, 斑马鱼行为强度平均值分别为0.68±0.09和0.64±0.09, 差异值分别为 7.6%和 18.64%, 与对照组相同, 光周期行为强度明显高于暗周期(P<0.01), 证明斑马鱼的昼夜节律性。(2)在氨氮胁迫下斑马鱼行为响应最显著的特点是行为活动遭到抑制。低浓度胁迫轻微抑制, 高浓度胁迫严重抑制。在有效氨氮浓度下, 浓度越高, 抑制作用越明显(P<0.001)。(3)斑马鱼行为响应具有显著的昼夜节律现象。Autocorrelation 中正负相关峰值交替性出现, 总体表现出较为良好的对称性和以 24h 循环的周期性, 通过SOM 分析, 大部分行为强度数据与光暗周期时间分布相匹配, 说明斑马鱼的昼夜节律现象。研究表明斑马鱼行为响应具有明显的昼夜节律现象, 光周期行为强度高于暗周期, 周期为 24h 左右; 在氨氮胁迫下斑马鱼行为强度遭到抑制, 在有效氨氮浓度下浓度越高, 抑制作用越明显, 出现时间节点延迟和聚类分析异常的现象。研究使用第三代在线生物行为监测系统, 其灵敏性和精确度显著提高, 能够实时准确反映氨氮胁迫下斑马鱼的行为响应变化, 也表明在线生物监测技术在水生生物行为学研究中发挥着不可或缺的作用。     

Immunological parameters of Javanese carp Puntius gonionotus (Bleeker) exposed to copper and challenged with Aeromonas hydrophila.

This study was to determine the median lethal concentration (LC50) of copper to Javenese carp, Puntius gonionotus (Bleeker), and the immune response after the fish were exposed to sublethal levels of copper and challenged with formalin killed Aeromonas hydrophila. The LC50 of copper on P. gonionotus at 24, 48, 72, 96 and 120 h were estimated as 2.17, 0.91, 0.57, 0.53 and 0.42 mg l(-1), respectively. To determine the effect of copper on the immune system, fish were exposed for 66 days to 0.05, 0.10 and 0.15 mg Cu l(-1). After 56 days of initial exposure to copper, fish were challenged with 0.1 ml of 4.5 x 10(5) cfu ml(-1) formalin killed A. hydrophila and maintained in the same concentration of copper. After the challenge, the immune response was monitored for 2 weeks using haematological and serological assays. During the initial phase of exposure to copper, significant changes were noted in the white blood cell, lysozyme, potential killing activity, total plasma protein, total immunoglobulin and haematocrit levels between the control and treated fish. One week after challenge with A. hydrophila, there was a significant increase in the values of white blood cells, total protein and total immunoglobulin compared to the values before the challenge. However, these values were not significantly different (P>0.05) between the control and the treated fish. In contrast, NBT and lysozyme assays exhibited a significant difference (P<0.05) in fish exposed to 0.10 mg Cu l(-1) (0.525 +/- 0.17; 24.42 +/- 3.35 x 10(2) micromg ml(-1)) and 0.15 mg Cu 1(-1) (0.536 +/- 0.19; 21.78 +/- 1.29 x 10(2) micromg ml(-1)) compared to the control (0.746 +/- 0.31; 30.73 +/- 5.42 x 10(2) micromg ml(-1)) after the bacterial challenge (day 61). There was however no significant difference (P>0.05) in NBT and lysozyme levels in fish exposed to lower level of copper (0.05 mg Cu l(-1)), suggesting the absence of immunosuppressive effects at lower level of exposure.     

Sublethal concentrations of waterborne copper induce cellular stress and cell death in zebrafish embryos and larvae

Copper is an essential ion that forms part of the active sites of many proteins. At the same time, an excess of this metal produces free radicals that are toxic for cells and organisms. Fish have been used extensively to study the effects of metals, including copper, present in food or the environment. It has been shown that different metals induce different adaptive responses in adult fish. However, until now, scant information has been available about the responses that are induced by waterborne copper during early life stages of fish. Here, acute toxicity tests and LC50 curves have been generated for zebrafish larvae exposed to dissolved copper sulphate at different concentrations and for different treatment times. We determined that the larvae incorporate and accumulate copper present in the medium in a concentration-dependent manner, resulting in changes in gene expression. Using a transgenic fish line that expresses enhanced green fluorescent protein (EGFP) under the hsp70 promoter, we monitored tissue-specific stress responses to waterborne copper by following expression of the reporter. Furthermore, TUNEL assays revealed which tissues are more susceptible to cell death after exposure to copper. Our results establish a framework for the analysis of whole-organism management of excess external copper in developing aquatic animals.     

Hepatic proteome sensitivity in rainbow trout after chronically exposed to a human pharmaceutical verapamil

Verapamil (VRP), a cardiovascular pharmaceutical widely distributed and persistent in the aquatic environment, has potential toxicity to fish and other aquatic organisms. However, the molecular mechanisms that lead to these toxic effects are not well known. In the present study, proteomic analysis has been performed to investigate the protein patterns that are differentially expressed in liver of rainbow trout exposed to sublethal concentrations of VRP (0.5, 27.0, and 270 μg/liter) for 42 days. Two-dimensional electrophoresis coupled with MALDI-TOF/TOF mass spectrometry was employed to detect and identify the protein profiles. The analysis revealed that the expression of six hepatic acidic proteins were markedly altered in the treatment groups compared with the control group; three proteins especially were significantly down-regulated in fish exposed to VRP at environmental related concentration (0.5 μg/liter). These results suggested that the VRP induce mechanisms against oxidative stress (glucose-regulated protein 78 and 94 and protein disulfide-isomerase A3) and adaptive changes in ion transference regulation (calreticulin, hyperosmotic glycine-rich protein). Furthermore, for the first time, protein Canopy-1 was found to be significantly down-regulated in fish by chronic exposure to VRP at environmental related levels. Overall, our work supports that fish hepatic proteomics analysis serves as an in vivo model for monitoring the residual pharmaceuticals in aquatic environment and can provide valuable insight into the molecular events in VRP-induced toxicity in fish and other organisms.     

Effects of waterborne copper or zinc on the osmoregulatory response of bluegill to a hypertonic NaCl challenge

1. Bluegill (Lepomis macrochirus) were exposed to copper (0.77 mg/1) or zinc (2.35 mg/1) for 7 days after which they were challenged by being placed in a 250 mM NaCl solution.2. Non-metal exposed controls started to die in the NaCl after 8 hr but those that had been exposed to copper survived at least 22 hr and some survived for at least 3 days. Zinc exposure also caused better survival in the hypertonic NaCl but not to the same extent.3. Blood plasma osmolality, chloride, and protein, as well as the hematocrit, muscle and liver water, and gill Na-K-activated ATPase were determined in fish before and after 8 hr in the NaCl.4. Metal exposure alone produced no significant effects on the variables measured except for a decline in plasma chloride in zinc-exposed fish.5. Both copper- and zinc-exposed fish exhibited less of an increase in chloride upon being challenged with the NaCl than did controls. Osmolality showed a similar effect from copper but not from zinc exposure. Metal exposure produced no other differences.6. All fish exhibited an elevated hematocrit but no change in plasma protein or tissue water when challenged. The elevated hematocrit is probably a non-specific stress response. The other data are consistent with the hypothesis that there was an influx of salt but little if any efflux of water while in the hypertonic NaCl.7. The data suggest indirectly that copper, and to a much lesser extent zinc, seemed to reduce the permeability of the gills to an influx of chloride in this stenohaline species.     

In vitro toxicity of bithionol and bithionol sulphoxide to Neoparamoeba spp., the causative agent of amoebic gill disease (AGD)

The objective of the present study was to evaluate the in vitro toxicity of bithionol and bithionol sulphoxide to Neoparamoeba spp., the causative agent of amoebic gill disease (AGD). The current treatment for AGD-affected Atlantic salmon involves bathing sea-caged fish in freshwater for a minimum of 3 h, a labour-intensive and costly exercise. Previous attempts to identify alternative treatments have suggested bithionol as an alternate therapeutic, but extensive in vitro efficacy testing has not yet been done. In vitro toxicity to Neoparamoeba spp. was examined using amoebae isolated from the gill of AGD-affected Atlantic salmon and exposing the parasites to freshwater, alumina (10 mg l(-1)), seawater, bithionol or bithionol sulphoxide at nominal concentrations of 0.1, 0.5, 1, 5 and 10 mg l(-1) in seawater. The numbers of viable amoebae were counted using the trypan blue exclusion method at 0, 24, 48 and 72 h. Both bithionol and bithionol sulphoxide demonstrated in vitro toxicity to Neoparamoeba spp. at all concentrations examined (0.1 to 10 mg l(-1) over 72 h), with a comparable toxicity to freshwater observed for both chemicals at concentrations > 5 mg l(-1) following a 72 h treatment. Freshwater remained the most effective treatment, with only 6% viable amoebae seen after 24 h and no viable amoebae observed after 48 h.     

The susceptibility of the giant freshwater prawn Macrobrachium rosenbergii to Lactococcus garvieae and its resistance under copper sulfate stress.

Addition of copper sulfate (0.1 to 0.4 mg l(-1)) to tryptic soy broth (TSB) had no effect on growth rate of the bacterial pathogen Lactococcus garvieae. Giant freshwater prawns Macrobrachium rosenbergii were injected with L. garvieae (4 x 10(6) colony-forming units [cfu] prawn(-1)) grown in TSB or TSB containing copper sulfate at 0.1, 0.2, 0.3 or 0.4 mg l(-1). After 48 h, the cumulative mortality was significantly (p < 0.05) higher for prawns exposed to L. garvieae grown in 0.4 mg l(-1) copper sulfate than at the lower concentrations examined. In other experiments, prawns were injected with TSB-grown L. garvieae (4 x 10(6) and 2 x 10(5) cfu prawn(-1)), then held in water containing copper sulfate. After 8 h the mortality of L. garvieae-exposed prawns held in water containing 0.4 mg l(-1) copper sulfate was significantly higher than prawns held in water containing 0.2 and 0.3 mg l(-1) copper sulfate. At the lower L. garvieae density, cumulative mortality of prawns increased directly with ambient copper sulfate concentrations in the range of 0.2 to 0.4 mg l(-1). All prawns survived a 168 h exposure to 0.1 mg l(-1) copper sulfate. Prawns exposed to different concentrations of copper sulfate were examined for hemocyte density, phenoloxidase activity and respiratory burst. No significant differences in hemocyte density were observed among treatments. In prawns following a 48 h exposure to 0.1 mg l(-1) copper sulfate, phenoloxidase activity was decreased, but respiratory burst was increased. In conclusion, copper sulfate increased the virulence of L. garvieae to M. rosenbergii and modulated its immune system. Copper sulfate at 0.1 mg l(-1) decreased susceptibility of M. rosenbergii to L garvieae infection, whereas at 0.2 mg l(-1) the susceptibility was increased. The generation of superoxide anion by M. rosenbergii exposed to copper sulfate at a concentration higher than 0.2 mg l(-1) was considered to be cytoxic.     

A review of boron effects in the environment

Boron (B) is a naturally occurring element that is found in the form of borates in the oceans, sedimentary rocks, coal, shale, and in some soils. Borates are released naturally into the atmosphere and aquatic environment from oceans, geothermal steams, and weathering of clay-rich sedimentary rocks. B is also released to a lesser extent from anthropogenic sources. B concentrations in air range from <0.5 to 80 ng/m³ with an average of 20 ng/m³, and in soils from 10 to 300 mg/kg with an average of 30 mg/kg. Concentrations of B in surface freshwaters are typically < 0.1–0.5 mg/L; much higher concentrations are measured in a few areas, depending on the geochemical nature of the drainage catchment. B accumulates in both aquatic and terrestrial plants, but it does not appear to be biomagnified through the food chain. No observed effect concentrations (NOECs) for aquatic invertebrates tend to be in the range of 6–10 mg B/L with lower values of 1–2 mg/L for community studies. No effect concentrations for fish in natural waters are around 1 mg/L, although lower values have been recorded in reconstituted water. Comparing no effect concentrations with the general ambient environmental levels indicates that the risk to aquatic ecosystems from B is low. In a few B-rich areas, natural levels will be higher; however, there is some indication that organisms may be Actapted to the local conditions. B is an essential micronutrient for higher plants with interspecies differences in the levels required for optimum growth. In general, there is a small concentration range between deficiency and toxicity; however, toxicity owing to excess B is much less common in the environment than B deficiency. Levels of B in aquatic plants growing in areas receiving B-rich runoff from irrigated fields are higher than dietary concentrations, which cause effects on the growth of young birds in the laboratory; however, the bioavailability in the field of such plant-accumulated B is uncertain.     

Oxidative Stress and Nano-Toxicity Induced by TiO2 and ZnO on WAG Cell Line

Metallic nanoparticles are widely used in cosmetics, food products and textile industry. These particles are known to cause respiratory toxicity and epithelial inflammation. They are eventually released to aquatic environment necessitating toxicity studies in cells from respiratory organs of aquatic organisms. Hence, we have developed and characterized a new cell line, WAG, from gill tissue of Wallago attu for toxicity assessment of TiO₂ and ZnO nanoparticles. The efficacy of the cell line as an in vitro system for nanoparticles toxicity studies was established using electron microscopy, cytotoxicity assays, genotoxicity assays and oxidative stress biomarkers. Results obtained with MTT assay, neutral red uptake assay and lactate dehydrogenase assay showed acute toxicity to WAG cells with IC₅₀ values of 25.29±0.12, 34.99±0.09 and 35.06±0.09 mg/l for TiO₂ and 5.716±0.1, 3.160±0.1 and 5.57±0.12 mg/l for ZnO treatment respectively. The physicochemical properties and size distribution of nanoparticles were characterized using electron microscopy with integrated energy dispersive X-ray spectroscopy and Zetasizer. Dose dependent increase in DNA damage, lipid peroxidation and protein carbonylation along with a significant decrease in activity of Superoxide Dismutase, Catalase, total Glutathione levels and total antioxidant capacity with increasing concentration of exposed nanoparticles indicated that the cells were under oxidative stress. The study established WAG cell line as an in vitro system to study toxicity mechanisms of nanoparticles on aquatic organisms.     

Similar stress responses are elicited by copper and ultraviolet radiation in the aquatic plant Lemna gibba: implication of reactive oxygen species as common signals

Metals and ultraviolet (UV) radiation are two environmental stressors that can cause damage to plants. These two types of stressors often impact simultaneously on plants and both are known to promote reactive oxygen species (ROS) production. However, little information is available on the potential parallel stress responses elicited by metals and UV radiation. Using the aquatic plant Lemna gibba, we found that copper and simulated solar radiation (SSR, a light source containing photosynthetically active radiation (PAR) and UV radiation) induced similar responses in the plants. Both copper and SSR caused ROS formation. The ROS levels were higher when copper was combined with SSR than when applied with PAR. Higher concentrations of copper plus PAR caused toxicity as monitored by diminished growth and chlorophyll content. This toxicity was more pronounced when copper was combined with SSR. Because the generation of ROS was also higher when copper was combined with SSR, we attributed this enhanced toxicity to elevated levels of ROS. In comparison to PAR-grown plants, SSR treated plants exhibited elevated levels of superoxide dismutase (SOD) and glutathione reductase (GR). These enzyme levels were further elevated under both PAR and SSR when copper was added at concentrations that generated ROS. Interestingly, copper treatment in the absence of SSR (i.e. copper plus PAR) induced synthesis of the same flavonoids as those observed in SSR without copper. Finally, addition of either dimethyl thiourea or GSH (two common ROS scavengers) lowered in vivo ROS production, alleviated toxicity and diminished induction of GR as well as accumulation of UV absorbing compounds. Thus, the potential of ROS being a common signal for acclimation to stress by both copper and UV can be considered.     

Acclimation of Daphnia magna to environmentally realistic copper concentrations

It may be hypothesised that as the bioavailable background concentration of an essential metal increases (within natural limits), the natural tolerance (to the metal) of the acclimated/adapted organisms and communities will increase. In this study the influence of acclimation to different copper concentrations on the sensitivity of the freshwater cladoceran Daphnia magna Straus was investigated. D. magna was acclimated over three generations to environmentally relevant copper concentrations ranging from 0.5 to 100 microg Cu/l (copper activity: 7.18 x 10(-15) to 3700 x 10(-12) M Cu2+). A modified standard test medium was used as culture and test medium. Medium modifications were: reduced hardness (lowered to 180 mg CaCO3/l) and addition of Aldrich humic acid at a concentration of 5 mg DOC/l (instead of EDTA). The effects of acclimation on these organisms were monitored using acute mortality assays and long-term assays in which life table parameters, copper body concentrations and energy reserves were used as test endpoints. Our results showed a two-fold increase in acute copper tolerance with increasing acclimation concentration for second and third generation organisms. Copper acclimation concentrations up to 35 microg Cu/l (80 pM Cu2+) did not affect the net reproduction and the intrinsic growth rate. The energy reserves of the acclimated daphnids revealed an Optimal Concentration range (OCEE) and concentrations between 5 and 12 microg Cu/l (0.5-4.1 pM Cu2+) and 1 and 35 microg Cu/l (0.023-80 pM Cu2+) seemed to be optimal for first and third generation daphnids, respectively. Lower and higher copper concentrations resulted in deficiency and toxicity responses. It was also demonstrated that up to 35 microg Cu/l, third generation daphnids were able to regulate their total copper body concentration. These results clearly indicate that bioavailable background copper concentrations present in culture media have to be considered in the evaluation of toxicity test results, especially when the toxicity data are used for water quality guideline derivation and/or ecological risk assessment for metals.     

Effects of Dietary Supplementation with Vitamin C and Vitamin E and Their Combination on Growth Performance,Some Biochemical Parameters,and Oxidative Stress Induced by Copper Toxicity in Broilers

This study investigated effects of dietary supplementation with vitamin C, vitamin E on performance, biochemical parameters, and oxidative stress induced by copper toxicity in broilers. A total of 240, 1-day-old, broilers were assigned to eight groups with three replicates of 10 chicks each. The groups were fed on the following diets: control (basal diet), vitamin C (250 mg/kg diet), vitamin E (250 mg/kg diet), vitamin C + vitamin E (250 mg/kg?+?250 mg/kg diet), and copper (300 mg/kg diet) alone or in combination with the corresponding vitamins. At the 6th week, the body weights of broilers were decreased in copper, copper + vitamin E, and copper + vitamin C + vitamin E groups compared to control. The feed conversion ratio was poor in copper group. Plasma aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase activities, iron, copper concentrations, and erythrocyte malondialdehyde were increased; plasma vitamin A and C concentrations and erythrocyte superoxide dismutase were decreased in copper group compared to control. Glutathione peroxidase, vitamin C, and iron levels were increased; aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and copper levels were decreased in copper + vitamin C group, while superoxide dismutase, glutathione peroxidase, and vitamin E concentrations were increased; aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase were decreased in copper with vitamin E group compared to copper group. The vitamin C concentrations were increased; copper, uric acid, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and malondialdehyde were decreased in copper + vitamin C + vitamin E group compared to copper group. To conclude, copper caused oxidative stress in broilers. The combination of vitamin C and vitamin E addition might alleviate the harmful effects of copper as demonstrated by decreased lipid peroxidation and hepatic enzymes.     

Biochemical and morphological changes in carp (Cyprinus carpio L.) liver following exposure to copper sulfate and tannic acid

As a consequence of human activity various toxicants reach the aquatic ecosystems; humics may interact with them and may change their toxicity. Many fish are exposed to a considerable concentration of humics and pollutants. Because of paucity of data on the biochemical action of tannins in the presence of the fungicide CuSO4 a comparative study was undertaken. The alterations of redox-parameters in carp liver were monitored and tissue necrosis was followed by measuring the plasma transaminase activities and by electron microscopy. Tannic acid, a representative phenolic/humic compound, exerted prooxidant effects in carp, which may be partially due to formation of prooxidant intermediates/end-products via its biotransformation. Alternatively, tannic acid may partially inhibit the antioxidant enzymes of fish. The response to CuSO4 was more severe. Although tannic acid alone acted as a prooxidant in fish, electron micrographs demonstrated that it reduced the necrotizing effect of copper, which may be due to the complexing activity of tannic acid with the biomolecules of the hepatocytes and to the H2O2-degrading activity of tannin-CuSO4 combination. Our results indicate that the heavy metal-detoxifying capacity of tannin may be significant; however, tannin-exposure alone or combined with metals may be toxic for fish due to enzyme inhibition and oxidative stress induction.     

Comparative study on the inhibition of acetylcholinesterase activity in the freshwater fish Cyprinus carpio by mercury and zinc.

The effects of sublethal concentrations of mercury (0.1mg/l) and zinc (6 mg/l) on acetylcholinesterase activity and acetylcholine content of gill, kidney, intestine, brain, liver and muscle of the freshwater fish Cyprinus carpio at 1, 15 and 30 days of exposure were studied. A significant suppression in acetylcholinesterase activity was recorded in all the organs from both mercury and zinc intoxicated fish at all the exposure periods. Concurrently, a significant increase in the content of acetylcholine in the organs was observed. These changes observed in the organs of mercury treated fish in different exposure periods were in the order 1 greater than 15 less than 30 days and in zinc treated fish 1 greater than 15 greater than 30 days. Further, these changes were greater in magnitude in the brain, liver and muscle (non-osmoregulatory organs) than in the gill, kidney and intestine (osmoregulatory organs) in both metal media.     

Induction of luteolysis in mares by ultrasound-guided intraluteal treatment with PGF2alpha.

To evaluate the technique of ultrasound-guided luteal injection in mares, PGF2alpha was administered under ultrasound guidance to horse mares (n = 7 to 9 per group) on Day 9 postovulation via either a systemic (i.m.; zero, 0.01, 0.1, or 5 mg/dose) route or a local intraluteal (i.l.; zero, 0.01 or 0.1 mg/dose) route. The luteolytic efficacy of each treatment was determined based on post-treatment decreases in progesterone concentration, interval to uterine edema (IE) and interovulatory interval (IOI). Local administration of PGF2alpha directly into the CL consistently induced luteolysis, at doses up to 50-fold lower than the lowest effective systemic dose. Significant decreases in IOI and IE occurred in mares treated with 5 mg PGF2alpha i.m. or 0.1 mg PGF2alpha i.l., but did not occur in mares treated with 0.1 or 0.01 mg PGF2alpha i.m., 0.01 mg PGF i.l., vehicle i.l. or vehicle i.m.. Progesterone concentrations were reduced to less than 10% of pretreatment values by two days post treatment in mares treated with 5 mg PGF2alpha i.m. or 0.1 mg PGF2alpha i.l.. PGF2alpha doses of 0.1 mg i.m. and 0.01 mg i.l. were associated with smaller but significant progesterone decreases (to 66% and 46% of pre-treatment values, respectively) by two days post treatment. Progesterone values after administration of i.l. vehicle did not differ from pre-treatment values by two days post treatment, but were significantly lower (53% of pre-treatment values) by four days post treatment. Intramuscular treatment with vehicle or 0.01 mg of PGF2alpha did not significantly reduce progesterone concentrations below pretreatment values. Overall, the minimum effective luteolytic dose of PGF2alpha given intraluteally was between 0.01 and 0.1 mg. Based on the results of this study, ultrasound-guided i.l. injection appears to be a repeatable method for studying the direct effect of other chemicals on luteal function. However, the current procedure carries some risk, since three i.l. injections were associated with ovarian abscesses.     

Aqueous fullerene aggregates (nC60) generate minimal reactive oxygen species and are of low toxicity in fish: a revision of previous reports

This review aims to clarify inconsistencies in previous reports regarding the potential for aqueous aggregates of fullerenes (nC60) to generate reactive oxygen species (ROS) and cause toxicity in fish. Methods for evaluation of ROS production and toxicity of aqueous nC60 have evolved over time and limitations in initial studies have led to unintentional erroneous reports of nC60 ROS generation and toxicity. Some of these reports continue to lead to misconceptions of the environmental effects of C60. Critical review of the evidence (2007-2011) indicates that aqueous nC60 have minimal potential to produce ROS and that oxidative stress in fish is not induced by environmentally relevant exposure to nC60. Future studies should acknowledge that current evidence indicates low toxicity of nC60 and refrain from citing articles that attribute toxicity in fish to nC60 based on methods shown to be compromised by experimental artifacts. Despite low toxicity of nC60 in fish, an emerging environmental issue is that nC60 can affect environmental fate, transport, and bioavailability of co-contaminants in aquatic environments in a similar manner to that observed for other anthropogenic particulates (e.g., microplastics).