Protective effect of Kappaphycus alvarezii (Rhodophyta) extract against DNA damage induced by mercury chloride in marine fish

Marine organisms are continuously exposed to agents, both exogenous and endogenous, that damage DNA. Consequently, it is important to determine the ability of compounds to provide protection against damaging chemicals. The aim of this study was to evaluate the anti-genotoxic activity of crude aqueous extracts of Kappaphycus alvarezii (Rhodophyceae), collected from the Southeast coast of India. This study focused on possible anti-genotoxic potential of aqueous extract of K. alverazii to interfere with clastogenicity induced by mercury chloride (HgCl₂) in marine fish, Therapon jarbua as measured by cytogenetic endpoints such as cell viability and comet assay. In the first set of experiments, fish were exposed to a single treatment of Hg at 0.125, 0.25, 0.5, 1, or 2?ppm along with controls. Mercury exposure produced significant DNA damage in all comet classes, maximum as >79% (Class 4) at 0.5, 1, and 2?ppm exposure in a time dependent manner. Algal extract did not induce genotoxicity when given alone and prevented Hg-induced genotoxicity. The algal extract reduction in genotoxicity was significant but not time- and concentration-dependent. Results suggested that under present experimental conditions, K. alvarezii extract exhibit potent anti-genotoxicity effects in this fish model; and thus these extracts may be recommended as a supplement in fish meal and may benefit humans ingesting Hg-contaminated fish.     

Mercuric chloride induced proteotoxicity and structural destabilization in marine fish (Therapon jarbua)

The reaction byproducts derived from lipid peroxidation (LPO), as well as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) enzyme activities were measured in tissues of marine fish (Therapon jarbua) exposed to mercuric chloride (HgCl) in water dispersions of 0.125 or 0.25 ppm. LPO was significantly increased in various tissues relative to control values after 96-h exposure. SOD and GPx activities significantly decreased after exposure to first two doses but significantly elevated CAT in Dose II kidney and liver tissues. The elevated levels of LPO, decreased activities of SOD and GPx, and increased CAT activities in all tissues examined in T. jarbua are an index of oxidative stress in fish. Structural analysis by scanning electron microscopy studies revealed the structural deformation in HgCl₂-exposed animals. These observations suggest that HgCl acts as a mediator in free radical generation. The increase in CAT and decrease in SOD and GPx activities in these tissues may be an adaptive response.     

十溴二苯乙烷对草鱼幼鱼肝脏和肌肉组织氧化应激效应的影响

十溴二苯乙烷(DBDPE)是目前在全球范围内广泛使用的新型溴代阻燃剂,其环境风险已引起广泛关注,但目前仍缺乏针对水生生物的毒性研究数据。作者通过饲料中添加十溴二苯乙烷暴露的方式对草鱼幼鱼进行长期暴露实验,研究500、1 000和3 000 mg·kg~(-1)三个饲料添加剂量暴露组和1个对照组长期暴露对草鱼幼鱼肝脏和肌肉组织中氧化应激酶(SOD、CAT和GSH-PX)活性和抗氧化物质(GSH)含量的影响。结果显示:暴露8周后,随着DBDPE暴露水平的升高,草鱼幼鱼肝脏组织中氧化应激酶(SOD、CAT和GSH-PX)和抗氧化物质(GSH)均表现出低浓度诱导及高浓度抑制的效应。500和1 000 mg·kg~(-1)剂量组草鱼幼鱼肝脏组织中SOD、CAT和GSH-PX活性和GSH含量均显著高于对照组(P0.05),且均在500 mg·kg~(-1)剂量组达到最高。3 000 mg·kg~(-1)剂量组SOD、CAT和GSH-PX活性和GSH含量低于500和1 000 mg·kg~(-1)暴露组,但与对照组无显著性差异(P0.05)。草鱼幼鱼肌肉组织中氧化应激酶活性变化甚微,3个浓度剂量组肌肉组织中SOD、CAT活性和GSH含量以及500 mg·kg~(-1)剂量组GSH-PX活性与对照组均无显著性差异(P0.05)。研究成果表明DBDPE暴露影响草鱼幼鱼肝脏组织的抗氧化防御系统,可以诱导草鱼幼鱼产生氧化应激效应。     

Effects of mercury graded doses on redox status,metallothionein levels and genotoxicity in the intestine of sea cucumber Holothuria forskali

Mercury (Hg) is a ubiquitous and a major environmental metal pollutant in the aquatic ecosystem. The present study was performed to evaluate the effect of mercury graded doses exposure on oxidative stress, redox status, metallothionein levels and genotoxicity in the intestine of sea cucumber Holothuria forskali. Specimens were exposed for 96?h to three concentrations of Hg (40, 80 and 160?µg/L). Exposure of H. forskali to Hg promoted oxidative stress with an increase in malondialdehyde (MDA), protein carbonyl (PCO) and advanced oxidation protein products (AOPP) levels. An increase of glutathione (GSH), vitamin C (ViteC) and non-protein thiols (NPSH) contents was also observed. Additionally, antioxidant activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) increased especially with the highest doses of Hg indicating a great defense of the antioxidant system. Our investigation revealed an increase in total Metallothionein (MT) content which was more pronounced at the sharpest concentrations of Hg. A significant decline of AChE activity was also observed. In the intestine of Hg-treated H. forskali genotoxicity was confirmed by DNA degradation.     

Histochemical demonstration of mercury in the intestine,nephridia and epidermis of the marine polychaete Nereis virens exposed to inorganic mercury

Marine polychaetes Nereis virens (Sars) were exposed to 9 ppb Hg as mercuric chloride in the aquarium water. Concentrations of mercury in the water and in the worms were monitored using radiolabelled mercury. After 11 d, the average mercury concentration in the nereids was 8.41 ppm Hg and the bioconcentration factor was calculated to be 930. Accumulations of mercury in the tissues were made visible for light and electron microscopy by autometallographic silver enhancement. Mercury was demonstrated in the intestine, nephridia, epidermis and cuticula. In the intestine, mercury was found to be located predominantly in the apical part of the epithelial cells. In the nephridia, mercury deposits were apparent in the peritoneal cells of the nephridial tubules. Silver-enhanced mercury was also observed at the epicuticula, in the collagen fibres of the endocuticula and in the epidermal basal cells. Intracellularly, the observed mercury accumulations were localised predominantly in lysosomes. Extracellularly, mercury was observed in the basal laminae of the intestine and the epidermis, and also in the intestinal peritrophic membrane. How N. virens copes with mercury toxicity is discussed.     

汞和砷单一及联合暴露对日本沼虾的氧化应激效应

为深入探讨汞(Hg)和砷(As)对水生生物的慢性毒性效应,以日本沼虾(Macrobrachium nipponense)为受试生物,以超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、金属硫蛋白(MT)和丙二醛(MDA)为测试分子标志物,研究上覆水中不同浓度Hg和As单一及联合暴露对日本沼虾肝胰腺的氧化损伤及联合作用机制。结果表明:Hg和As单一及联合暴露对4种分子标志物均可产生不同程度的抑制或激活效应。其中,SOD活性均受到抑制,且高浓度As(1000μg·L-1)暴露10 d的抑制率最大,高达59.13%;而CAT活性整体被激活,Hg单独暴露下的激活效应要高于As,且在联合暴露组中随着暴露时间的增加呈现出显著的剂量效应关系(P0.05);MT和MDA含量在Hg和As的暴露下均显著增加(P0.05)。其中MT对Hg的螯合能力大于As,As单独暴露3 d后MDA变化较小,但10 d后其含量显著增加(P0.05)。析因方差分析发现,Hg-As对机体的联合作用机制主要表现为协同作用。综合生物标志物响应指数(IBR)评价证实,联合毒性要高于单独毒性,Hg与As在高浓度下联合暴露10 d的IBR值最大,毒性最强,高浓度As单独暴露处理组的毒性次之;同时,机体自身随暴露时间延长对Hg也表现出一定的解毒功能。     

The microcystins-induced DNA damage in the liver and the heart of zebrafish,Danio rerio

Microcystins (MCYST) are the freshwater cyanobacterial toxins, known to induce hepatocellular carcinoma, necrosis, intrahepatic bleeding, as well as human and livestock mortality. Within hepatocytes, MCYST selectively bind to protein phosphatases 1 and 2A, resulting in severe liver damage. The toxicology of MCYST in mice and rats has been well studied, but little is known regarding genotoxicity in aquatic animals. In this study, the zebrafish, Danio rerio was exposed to crude extract of Microcystis aeruginosa bloom. Liver and heart were examined for MCYST-induced toxicity. Light microscopy at 36?h revealed severe, widespread apoptotic necrosis of the majority of hepatocytes, and cytoskeletal deformation in myocardiocytes. Hepatocytes were dissociated with cell shrinkage and margination of nuclear chromatin. Laddering of genomic DNA from the liver and heart of the exposed fish in an increment of 180–200?bp was consistent with apoptosis. Fluorimetric analysis of DNA unwinding was carried out to determine the DNA strand breakage. After 36?h exposure, the % double-stranded DNA was significantly reduced in hepatocytes and myocardiocytes. In conclusion, the results obtained in this study indicate that, the extract of M. aeruginosa bloom is genotoxic to fish. The DNA damage observed in this study may be attributed to the activation of DNA endonucleases. This model of DNA damage may contribute for identifying novel molecular mechanisms of interest for therapeutic application.     

汞、硒暴露对紫贻贝(Mytilus edulis)抗氧化酶系统的影响

为了研究重金属汞及微量元素硒对海洋贝类的毒性效应,揭示汞、硒在生物体内的相互作用机制,用汞和硒对指示生物紫贻贝(Mytilus edulis)进行单一及联合亚慢性暴露实验。设置对照组(0μg·L-1)、汞暴露组(25μg·L-1Hg2+)、硒暴露组(4μg·L-1Se4+)以及硒汞联合暴露组(25μg·L-1Hg2++4μg·L-1Se4+)4个实验组,并分别在暴露期间的第0、2、4和6天定时采集样本,测定紫贻贝鳃SOD、GPx及CAT3种抗氧化酶活性。将实验数据进行ANOVA分析处理后,结果表明:与对照组相比,汞暴露组SOD和GPx均呈现先显著升高(p<0.05或p<0.01)后降低(p<0.01或p<0.001)的趋势,CAT则从第4天开始显著降低(p<0.05);硒暴露组中SOD活性始终高于对照组(p<0.001),GPx活性在第2天也显著升高(p<0.001),CAT活性始终与对照组相近;硒汞联合作用与汞暴露组相比,SOD、GPx和CAT活性在不同时间点均有显著升高(p<0.05),而与硒暴露组相比,3种酶活性均低于硒单独暴露的水平。说明汞在短期能够诱导抗氧化酶活性,随着暴露时间的延长,则表现出明显的抑制作用;微量硒能够增强抗氧化酶系统活性,对汞导致的氧化损伤具有拮抗作用。     

Radical scavenging activity of Trianthema triquetra in male albino rats intoxicated with CCl4

Radical scavenging activity of ethanolic extract of Trianthema triquetra root was investigated against CCl4 in rats. The rats were treated with T. triquetra (100 mg, 200 mg/kg b.w.) for a period of 7 days. Antihepatotoxic effect was studied by assaying the activities of thiobarbituric acid (TBARS), reduced glutathione (GSH), glutathione peroxidase (GPx),catalase (CAT), super oxide dismutase (SOD) and vitamin C (Vit. C). Lipid peroxidation is evidenced by an increase in the value of TBARS and also a distinct diminution in the level of GSH, Vit. C at 200 mg/kg b.w. The activity of antioxidant enzymes, such as GPx, CAT SOD and Vit. E is significantly recovered towards an almost normal level in animals coadministrated with T. triquetra. The maximum protection against CCl4 induced hepatic injury was afforded by the dose of 200 mg/kg b.w. of Trianthema triquetra.     

Biomarkers of oxidative stress as indicators of water pollution in Nile tilapia (Oreochromis niloticus) from a water reservoir in Riyadh,Saudi Arabia

Antioxidant enzyme activities of fish (Oreochromis niloticus) were determined in order to assess the status of pollution in the Wadi Namar (WN), near Riyadh, Saudi Arabia. Activities of four antioxidant enzymes as superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and concentrations of glutathione (GSH) and oxidant malondialdehyde (MDA) were selected as bioindicators. Fish (n = 14) were sampled in the month of April 2013 from WN and a control site (CS). SOD activity was increased by 37.9%, 47%, and 29% in kidney, liver, and heart, respectively, while a significant decrease (36.4%) was observed in gills of O. niloticus from WN as compared to fish from CS. CAT activity was reduced by 51%, 55%, 47%, and 35% in kidney, liver, heart, and gills of O. niloticus from WN. The GST activities in kidney, liver, and heart of O. niloticus from WN were elevated by 34%, 48%, and 32%, respectively. However, significant fall (49%) in gills of fish was noted from WN compared to fish from CS. GSH levels were increased by 44%, 36%, and 38% in kidney, liver, and heart, respectively, but decreased by 30% in gills. MDA levels of O. niloticus were significantly increased in kidney, liver, and heart in fish from WN. Data indicated that WN is polluted mainly by industrial and urban discharge of liquid waste products.     

Assessment of individual and combined toxicities of three heavy metals (Cu,Cd and Hg) by using Tigriopus fulvus

The toxicities of copper, cadmium, and mercury ions and their binary and ternary mixtures were studied using the copepods Tigriopus fulvus. The LC50 values measured after 48 h exposure to single metal solutions revealed a toxicities rank as follows: CdT. fulvus. The combined effect of the metals was found to be antagonistic for Cu?Cd, Hg?Cd, and Cu?Cd?Hg, additive for Hg?Cu.     

乙酸铜对银鲳幼鱼急性毒性及抗氧化酶活性的影响

为评价乙酸铜对银鲳的安全性及毒性效应,采用静水急性暴露实验,研究了8个乙酸铜浓度梯度(0、0.150、0.206、0.282、0.387、0.531、0.729、1.00 mg·L~(-1))对银鲳幼鱼的急性毒性,以肝脏和鳃组织的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化酶(GSH—PX)以及谷胱甘肽巯基转移酶(GST)活性为指标研究乙酸铜对银鲳幼鱼的毒理作用,并进行安全评价。结果表明,高浓度乙酸铜会对银鲳幼鱼产生了较大的毒性,48 h和96 h-LC_(50)分别为0.898 mg·L~(-1)、0.264 mg·L~(-1),安全质量浓度为0.026 mg·L~(-1);不同浓度乙酸铜胁迫下银鲳幼鱼组织中的SOD、CAT、GSH-PX和GST活性均表现为低浓度被诱导而高浓度受抑制的规律,与乙酸铜浓度呈抛物线型剂量效应关系,此外,肝脏中4种抗氧化酶活性普遍高于鳃组织。研究发现,乙酸铜胁迫对抗氧化酶的影响,可以反映银鲳幼鱼机体的受损状况,其中SOD可较灵敏地指示早期低浓度的铜污染。     

Hepatoprotective effects of taurine against mercury induced toxicity in rats

An attempt has been made to study the influence of taurine on mercury intoxicated rats. The animals were treated with sublethal dose of mercuric chloride (2 mg/kg body wt.) for 30 days. During the mercury treatment, the level ofAspartate transaminase(AST), Alanine transaminase (ALT) and Alkaline phosphatase(ALP) in serum and lipid peroxidation (LPO) in liver tissue significantly increased whereas Glutathione (GSH), Glutathione peroxidase(GPx), Catalase (CAT) and Superoxide dismutase (SOD) were simultaneously decreased in the liver tissue. Present results indicate that the liver tissue was completely damaged, after mercury treatment. In another group of animals, taurine (5 mg/kg body wt.) was administrated for another 15 days. Taurine administration was observed to improve the liver function in mercury intoxicated animal as indicated by the decline in increased levels of AST, ALT and ALP in serum and LPO content in liver tissue. The decreased level of antioxidant system (GSH, GPx, CATand SOD) has been promoted Results suggested that taurine played a vital role in reducing the mercury toxicity in intoxicated animals.     

Antioxidant enzymatic activities and lipid peroxidation in liver and ovary of zebrafish (Danio rerio) exposed to deltamethrin

Deltamethrin (DM) is being used as a substitute for organochlorines and organophosphates in pest control because of its low environmental persistence and toxicity. But it has become an environmental contaminant as it has been used widely. In this study, we investigated the effect of DM (technical grade) on the antioxidant system of adult zebrafish. For this, six-month-old fish were exposed to 2, 4 and 6?μg/L of DM for 96?h. The tissues selected were liver and ovary. Our data showed that exposure to DM increases CAT (catalase), SOD (superoxide dismutase), GPx (glutathione peroxidase, antioxidant enzymes), LPO (lipid peroxidation, non-enzymatic antioxidant) and GST (glutathione S-transferase, detoxifying enzyme) in liver and ovary. Increased GST could detoxify the toxicant; still there could be enough DM to cause oxidative stress. It appears from our study that zebrafish used compensatory mechanisms in eliminating reactive oxygen species. These data will be useful as oxidative stress is being used as a biomarker for aquatic pollution.     

Oxidative stress and changes in locomotor behavior and gill morphology of Gambusia affinis exposed to chromium

Sublethal effects of chromium trioxide on mosquito fish, Gambusia affinis, were carried out for 28 days on day 4, 8, 12, 16, 20, 24, and 28. The sublethal effects of chromium (LC₁₀, 77.62?mg?L^?1) on antioxidant enzymes, thiobarbituric acid reactive substance (TBARS), locomotor behavior, and gill morphology were studied. The antioxidant enzymes like superoxide dismutase (SOD) and catalase were induced throughout the exposure span of 28 days. Percent induction was also increase with the days of exposure. Lipid peroxidation product, malondialdehyde was enhanced in the viscera tissue of chromium-exposed fish. Perturbation of locomotor behavior like decrease in distance travelled (m?min^?1) and in swimming speed (cm?s^?1) was observed in exposed fish. Chromium toxicity caused alterations in gill morphology like hypertrophy and hyperplasia in secondary lamellae followed by detached epithelium with severe necrosis. It can be concluded from the results that Cr intoxication in Gambusia fish, caused induction in antioxidant enzymes and lipid peroxidation, plus changes in gill morphology and locomotor behavior.     

Total mercury in marine species from the French coast of the Eastern English Channel

Mercury (Hg) in fish can present a potential risk to wildlife and human consumers. Mercury levels were determined in 12 fish species and 2 invertebrate species from the French coast of the Eastern English Channel collected in June and November 2012. Total mercury concentrations in the muscle tissues ranged between 0.04 and 1.63 µg g^-1 (d.w.). Dab and sandeel displayed a significant positive correlation between Hg concentrations and fish length. Seasonal differences in Hg concentrations were observed for dab and shrimp. The lesser weever showed significantly higher concentrations of Hg (0.97 ± 0.34 µg g^-1 d. w.) than all other species. Gobies and sprat, which had the smallest size, showed significantly lower concentrations than herring, whiting, pout and sardine. Among the considered species, flatfish such as dab, flounder, plaice and sole displayed intermediate levels of Hg. In the present study, the fish age expressed as specimen length seems to be one of the major factors governing Hg contents in muscle tissues. Finally, mercury concentrations in commercial fish from the Eastern English Channel were below the levels fixed by the European Union for total mercury in edible parts of fish products.     

Total and methyl mercury in small marine biota caught off the Coast of Chennai,India

Abstract

This study reports the concentrations of total mercury and methyl mercury in sixteen families of marine biota caught off the coast of Kasimedu in Chennai, India, an important but understudied fish landing region. These included the commonly caught croakers, carangids, rays, goat fish, anchovies, crabs, and prawns. There was no correlation between total mercury or methyl mercury with fish length or mass. All concentrations were lower than the Food Safety and Standards Authority of India limits (total mercury = 500?μg/kg; methyl mercury = 250?μg/kg). Some values were above screening levels (total mercury >40?μg/kg wet weight) when considering possible adverse effects in predatory fish that consume the analyzed biota.

Abbreviations: dw: dry weight; MeHg: methyl mercury; THg: total mercury; ww: wet weight     

Effects of Matricaria chamomilla L. on lipid peroxidation,antioxidant enzyme systems,and key liver enzymes in CCl4-treated rats

In this study, we investigated the effects of Matricaria chamomilla L. extract (MCE) on lipid peroxidation, antioxidant enzyme systems, and several liver enzymes in carbon tetrachloride (CCl₄)-treated rats. Rats were divided into five groups. The first group (control group) was fed on standard feed. The rats in the other groups (CCl₄, MCE50, MCE100, and MCE200) were injected intraperitoneally with 0.8?mL?kg^?1 CCl₄. Moreover, rats in the MCE50, MCE100, and MCE200 groups were gavaged with 50?mg?kg^?1, 100?mg?kg^?1, and 200?mg?kg^?1 MCE, respectively. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, whole blood malondialdehyde (MDA) and glutathione (GSH) levels, and erythrocyte superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activity levels were measured after 14 days of exposure. ALT and AST in the CCl₄ group increased significantly in comparison to the control group (p?4, MCE50, MCE100, and MCE200 groups at different significance levels. In conclusion, the findings suggest that, depending on the dose administered, MCE decreases CCl₄-induced damage and consequent oxidative stress in rats; it affects the antioxidant system positively.     

Histology of Sculpin spp. in East Greenland. II. Histopathology and trace element concentrations

Abstract

For many years, the sculpin has been utilized as a sentinel monitoring species for anthropogenic impacts on the marine environment. To further develop its potential as a screening model body burden of several trace elements, including cadmium (Cd), copper (Cu), mercury (Hg), lead (Pb) and selenium (Se), were investigated. In addition, histopathology in shorthorn sculpins (Myoxocephalus scorpius; n?=?20) and fourhorn sculpins (Myoxocephalus quadricornis; n?=?10) were examined at three sites around the Scoresby Sound settlement in Central East Greenland. Fourhorn sculpins caught at the most distant site from the settlement, contained significantly higher hepatic Cu and Cd concentrations as well as significantly elevated gill Pb levels compared to the shorthorn sculpins collected at two other sites closer to the settlement. Histological examinations showed that fish with significantly higher concentrations of hepatic Cd and Se exhibited greater nuclear alterations, interstitial hyperplasia/hypertrophy, interstitial mononuclear infiltrations and granulomas. Further, fish with higher Cd and Cu gill tissue levels displayed a significantly higher number of cytoplasmic alterations and lamellar epithelium lifting, hypertrophic and hyperplastic epithelium along with mucus cell hyperplasia. While the presence of liver lesions were not species or sex-specific, the presence of gill lesions decreased in the order female fourhorn sculpins?>?female shorthorn sculpins?>?male fourhorn sculpins?>?male shorthorn sculpins. Hepatic Hg concentrations exceeded known lowest observable effect doses (LOED) for fish (0.1–0.5?μg g^?1 ww) in 27% of fish, while liver Cd residues in 80% exceeded LOED (0.42–1.8?μg g^?1 ww). Based upon these results, data suggest that using the sculpin as a valuable sentinel fish species histopathology may serve as a reliable tool for assessing marine ecosystem exposure to trace metals. However, confounding physiological and ecological factors also need to be considered.     

Identification of protein biomarkers of mercury toxicity in fish

Bioaccumulative metals such as mercury are found in increasing amounts in fish and their consumers. In the region of the Madeira River, in the Brazilian Amazon, mercury (Hg) is a predominant contaminant in the aquatic ecosystem. There is therefore a need to find specific biomarkers of mercury toxicity in fish to monitor contaminations. Here, mercury-bound proteins were identified in the liver tissues of fishes Mylossoma duriventre and Brachyplatystoma rousseauxii. Mercury was quantified in liver tissue, pellets and protein spots by graphite furnace atomic absorption. Proteins were fractionated by two-dimensional polyacrylamide gel electrophoresis and identified by mass spectrometry with electrospray ionization. We identified nine proteins linked to mercury and that presented biomarker characteristics of mercury. Among the proteins identified, isoforms of parvalbumin, ubiquitin-40S ribosomal protein S27a, brain-specific angiogenesis inhibitor 1-associated protein 2-like protein 2 and betaine–homocysteine S-methyltransferase 1 are notable for having the molecular function of binding to metallic ions.