Influence of marine alga against mercury-induced biochemical changes,structural destabilization,and genotoxicity in marine tiger fish,Therapon jarbua

This study elucidated the protective role of alga against mercury (Hg)-induced toxicity in marine fish. Bath immersion with mercury chloride (HgCl) [0.125 and 0.25?ppm] in combination with the algal extract (3?ppm) to Therapon jarbua offered a significant protection against Hg only induced disturbed liver function, damaged histoarchitecture, elevated oxidative stress, and DNA fragmentation of tissues. Mercury exposure decreased hepatic superoxide dismutase (SOD), catalase (CAT) activities, and the level of nonprotein-soluble thiol (NPSH, GSH), with a concomitant increase in thiobarbituric acid reactive substances (TBARS) in the gill, kidneys, liver, and blood. Algal supplementation diminished the rise in TBARS restoring CAT, SOD, and GSH levels to control. Reduced generation of free radicals may be correlated to protect DNA stability and morphology. This study indicates the ability of alga to ameliorate Hg-mediated injuries. In conclusion, Kappaphycus alvarezii extract did not show any toxicity and its safety is suggestive for using as a supplement in fish food.     

Oxidative stress and genotoxicity of 1-methyl-3-octylimidazolium chloride on loach (Misgurnus anguillicaudatus)

This study was a preliminary step to evaluate the acute toxicity of 1-methyl-3-octylimidazolium chloride ([C₈mim]Cl) on loach (Misgurnus anguillicaudatus) by determining the effects on hepatic antioxidant enzyme activities and by the comet assay. The results showed that [C₈mim]Cl had acute toxicity at concentrations above 20 mg L^?1, inducing oxidative stress and genotoxicity on fish liver cells. In respect to enzyme activities, [C₈mim]Cl induced changes in the activities of superoxide dismutase, catalase, and glutathione content the livers of fish exposed at 20–80 mg L^?1. [C₈mim]Cl at the same exposure level caused a remarkable increase in malondialdehyde level. The comet assay indicated that [C₈mim]Cl at 20–80 mg L^?1 induced genotoxicity in liver cells. With increased exposure concentration and time, the two comet parameters trailing rate and tail moment were significantly increased, with significant differences (P < 0.05) observed between control group and each treatment group. The present study shows that ionic liquids can be a threat to the health of aquatic organism when accidentally released to aquatic ecosystems.     

人血淋巴细胞检测浊漳河地表水的遗传毒性

为评价浊漳河水体中有机污染物的致突变性,采用单细胞凝胶电泳(SCGE)技术,研究了浊漳河水体3个不同样点的有机提取物对人外周血淋巴细胞DNA损伤效应。结果显示,各水样中的有机提取物均能引起DNA损伤,且随剂量的增高,有机物对DNA的损伤加重;细胞DNA损伤与对照组相比均有显著性差异(p<0.01)。这表明人外周血淋巴细胞对水体有机污染物的毒作用非常敏感。细胞彗星尾部DNA含量(%DNAT)和染毒剂量呈极显著正相关(R2>0.8547,p<0.01)。以上结果表明,浊漳河水体受到一定程度的有机物污染,这些有机污染物可诱导淋巴细胞DNA损伤,具有遗传毒性。SCGE技术作为一种简便、快速和灵敏的检测方法在水环境遗传毒性监测方面具有较大的应用价值。     

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.     

Different patterns of carotenoid composition and photosynthesis acclimation in two tropical red algae

To be able to survive, marine macroalgae in shallow coastal waters need mechanisms for short-term acclimation to fast changes in their environment. Of major importance are mechanisms that regulate the efficiency of photosynthesis by protecting PS II from photo-oxidative damage. Carotenoids, xanthophyll cycles and non-photochemical quenching (NPQ) are central constituents of such protection mechanisms. Red algae as a group do not have a universal carotenoid composition. We screened ten red algal species and selected two species, originating from similar ecological conditions but with different carotenoid compositions, for use in irradiance-acclimation experiments. We selected the tropical intertidal species Gracilaria domingensis and Kappaphycus alvarezii with antheraxanthin and lutein as major xanthophylls, respectively. Simultaneous in vivo fluorescence and O₂ evolution experiments were performed at different irradiance levels, which allowed a direct comparison of overall photosynthetic performance with NPQ. Interconversions of xanthophylls (violaxanthin, zeaxanthin, β-cryptoxanthin and one unidentified carotenoid) did occur in G. domingensis, but not in response to sudden exposure to light. Thus, NPQ was not correlated with any xanthophyll cycle during short-term acclimation to light. G. domingensis had five times higher weight-specific photosynthetic rates than K. alvarezii, which can be explained by the thicker thallus of K. alvarezii. Chlorophyll-specific gross photosynthetic rates were higher in K. alvarezii, but net rates were the same for both species. G. domingensis showed an immediate strong onset of NPQ upon exposure to irradiance, followed by downregulation to the NPQ level required. In K. alvarezii NPQ increased slowly until the required NPQ level was reached. At high irradiance G. domingensis downregulated photosynthesis while K. alvarezii continued to produce O₂ even at 2,000 μmol photons m⁻² s⁻¹ without NPQ increase. The strategy of K. alvarezii may provide short-term gains but with the risk of oxidative damage. The fast onset of NPQ in G. domingensis even at subsaturating irradiance as well as downregulation of photosynthesis when NPQ is saturated might provide this species with a competitive advantage under conditions of changing irradiance in the field.     

Relationship between photosynthesis and non-photochemical quenching of chlorophyll fluorescence in two red algae with different carotenoid compositions

Algae are continuously exposed to short-term fluctuations in irradiance. We investigate how two red algae species regulate photosynthetic efficiency to cope with such changes and identify some strategies that differ from higher plants. Two red algae, Gracilaria domingensis and Kappaphycus alvarezii, with antheraxanthin and lutein as major xanthophylls, respectively, reacted to the onset of low light (below E _k) with a substantial decrease of NPQ. This is different from higher plants, but similar to previous observations in, e.g. cyanobacteria where it indicates an increase in the effective absorbance cross-section of Photosystem II (PSII) by state transition. Kinetic studies in continuous light revealed a high susceptibility of PSII to light stress ((1-q _P)/NPQ) in K. alvarezii immediately after the sudden onset of high light, followed by a decrease. This was caused by a slower onset of NPQ in K. alvarezii, followed by acclimation. In G. domingensis, susceptibility of PSII to light stress was stable with time, but absolute values of (1-q _P)/NPQ were higher than in K. alvarezii. These observations suggest that K. alvarezii may be better adapted to high light levels, but is less well prepared for large sudden changes in irradiation. In K. alvarezii, photosynthesis continued to increase with increasing irradiation when NPQ was saturated. As (1-q _P) and NPQ were still balanced in this situation, most likely, processes other than photosynthetic oxygen release are responsible for the increasing net O₂ production observed.     

Genotoxic assessment of anthracene and benzo [a] pyrene to milkfish Chanos chanos

Mutagenic and genotoxic effects of polycyclic aromatic hydrocarbons, anthracene and benzo [a] pyrene (BaP), in milkfish Chanos chanos were determined using micronucleus (MN) test and comet assay (CA). Distinct mean frequencies of nuclear abnormalities such as MNs; binucleated micronuclei, nuclear bud, and fragmented apoptotic cells were measured. Significant increase in DNA damage with five classes of damage level was observed and expressed in terms of arbitrary unit (AU). Mean frequencies of total nuclear abnormalities were 0.5?±?0.25 cells in control; 0.67?±?0.33 cells in solvent control; 70?±?9.60 cells in 0.176?mg?L^?1 anthracene, and 91.83?±?6.25 cells in 0.031?mg?L^?1 BaP. The greatest DNA damage of 170AU was observed in 0.176?mg?L^?1 anthracene-exposed group and 182AU was observed in 0.031?mg?L^?1 BaP-treated fish. This study confirmed that the CA and MN assays are useful tools in determining potential genotoxicity of water pollutants and might be appropriate as a part of monitoring program.     

彗星试验检测地下水有机污染物的遗传毒性效应

应用人外周血淋巴细胞和小鼠淋巴细胞彗星试验,对长治市5个样点的地下水有机浓集物进行了遗传毒性研究。结果表明,各水样中有机浓集物在3个染毒剂量50,200,800 m L·管-1条件下均可对人外周血淋巴细胞和小鼠淋巴细胞DNA产生不同程度的遗传损伤,并且随着浓度的升高,DNA损伤程度明显增加,各实验组与溶剂对照组相比有P<0.05或P<0.01显著差异。研究表明,上述2种淋巴细胞彗星试验可有效地检测地下水体有机污染物的遗传毒性。     

Toxicological and larvicidal activities of Alzanzalakhet,Melia azedarach against cucurbit fly,Dacus ciliatus at Hail Province in Saudi Arabia

Alzanzalakhet, Melia azedarach L. (Sapindales: Meliaceae) is a common medicinal plant which is found in Hail desert in Saudi Arabia and traditionally used for various purposes. In this study, toxin effectiveness of the ethanolic, acetonic, and aqueous extracts of M. azedarach leaves was tested to determine a larvicidal activity against the cucurbit fly, Dacus ciliatus (Loew) (Diptera: Tephritidae). Different treatments with M. azedarach extracts varied significantly in their impact on mortality of D. ciliatus larvae. Results showed that all tested extracts in the three solvents exerted a positive pattern for reducing larvae of D. ciliatus. However, the ethanolic M. azedarach extract was the most potent compared with other tested solvents. The highest mortality of the D. ciliatus larvae was shown to be 94%, 74%, and 68% when the concentration was 1?×?10⁵?ppm for ethanolic, acetonic, and aqueous extracts, respectively. In contrast, the concentration 1?×?10¹?ppm resulted in the lowest mortality of D. ciliatus larvae at 40%, 34%, and 12% for ethanolic, acetonic, and aqueous extracts, respectively. Overall, the results of this study suggest that the ethanolic extract of M. azedarach leaves may be used as a sustainable method for controlling D. ciliatus larvae.     

石油烃对栉孔扇贝血淋巴细胞DNA损伤的初步研究

为研究石油烃对海洋生物的毒性效应,将栉孔扇贝(Chlamys farreri)暴露于0.08、0.21和0.88mg·L-1石油烃中,采用单细胞凝胶电泳实验(彗星实验)技术检测不同暴露时间扇贝血淋巴细胞的DNA损伤程度,对照组中石油烃背景浓度为0.04mg·L-1。结果显示,低浓度(0.08mg·L-1)的石油烃短期(<7d)内即可导致栉孔扇贝血淋巴细胞的DNA损伤,并且随石油烃浓度的增大和暴露时间的延长,DNA损伤程度增加,石油烃浓度达0.88mg·L-1时,DNA损伤程度已非常严重。3d恢复实验后,各浓度组DNA损伤又均有不同程度的恢复。研究表明,彗星实验是检测石油烃对海洋贝类DNA损伤的一种有效手段,贝类血淋巴细胞DNA损伤有望成为石油烃污染的一种生物标志物,用于海洋污染的早期预警监测。     

Effect of supplemented soybean (Glycine max L) diet and extracts on aluminum sulfate-induced genotoxicity

Aim of this study is to evaluate the protective role of Glycine max L. (soybean) against aluminum sulfate-induced genotoxicity. A diet containing 30% soybean and extracts obtained with various organic solvents were administered orally to male white Swiss mice prior to intraperitoneal injection of Al₂(SO₄)₃. Light microscopy was used for the examination of genotoxicity involving somatic cells, germ cells, and sperms. The main secondary metabolites contained in each extract were isolated and identified by chromatographic and spectral analyses. Administration of soy diet and extracts to the examined mice induced significant anti-mutagenic activity against bone marrow aberrations and spermatocyte and sperm abnormalities. At the end of this study, maximum protection was achieved using the methanolic extract (83% and 79%) followed by those obtained with chloroform (71% and 49%) and n-hexane (51% and 39%) for bone marrow aberrations and spermatocyte abnormalities, respectively. The observed activity of soybean extracts might be related to the main constituents isolated and identified. These main constituents were the isoflavone glycosides genistin and daidzin in addition to saponins (methanol extract), lignan matairesinol (chloroform extract), and β-sitosterol, unsaturated fatty acids as well as phospholipids (n-hexane extract). The results illustrated that soy extracts can be considered as alternative anti-mutagenic nutraceuticals and promising source for developing anti-mutagenic drugs.     

Effect of the chemical composition of organic extracts from environmental and industrial atmospheric samples on the genotoxicity of polycyclic aromatic hydrocarbons mixtures

Particulate organic matter (PM) present in the atmosphere is a complex mixture of chemicals like polycyclic aromatic hydrocarbons (PAH) that may exert adverse health effects including respiratory and cardiovascular disturbances and cancer. In this study, airborne samples from environmental or industrial areas exhibiting different physicochemical composition were compared for their capacities to induce DNA damage in human hepatocytes HepG2. DNA strand breaks and DNA adducts formed by benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE), the most reactive metabolite of the carcinogenic benzo[a]pyrene (B[a]P), were measured with the comet assay and by high-performance liquid chromatography (HPLC)/mass spectrometry, respectively. Cells were exposed to organic matter extracted from PM. Experiments were performed either at a constant concentration of B[a]P or at concentrations corresponding to fixed air volumes. Results show that industrial extracts tend to produce more benzo[a]pyrene diol epoxide-N ²-2′-deoxyguanosine (BPDE-N ²-dGuo) DNA adducts than strand breaks, whereas the opposite was observed with environmental extracts. The chemical composition of the extracts significantly impacts the nature and levels of DNA damage. The amount of B[a]P and interaction with other contaminants in the extracts need to be considered to explain the formation of DNA damage. These results emphasize the use of in vitro tests as promising and complementary tools to widely used toxic equivalent factor (TEF) approach in order to assess health hazards related to chemical exposure of the general population.     

Effect of copper and cadmium on carbon assimilation and uptake of metals by algae

Two species of blue green algae Spirulina platensis and Anacystis nidulans grown in artificial aqueous media were treated with Cu and Cd in concentrations of 0.01, 0.1, 1.0 and 10 ppm to study carbon assimilation and Chlorophyll (Chl) A content. The species were treated with concentrations of 0.1, 0.5, 1.0, 5.0 and 10.0 ppm to study the uptake of metals with exposure time. Carbon assimilation and Chl A content showed responses proportional to the concentration in the general form y = K + n ln C, where C is the concentration of metal in ppm, while in case of uptake the relation was y = KC”; (where C is the molar concentration x 10^‐6 of the metal). The n values in case of uptake was found to be < 1 indicating a non‐Langmuir type of sorption. The concentration factors of metals decreased with metal concentration in the medium.     

Effects of UV-B radiation on the gills of Catla catla during early development

Scanning electron microscopic (SEM) study of gills of Catla catla catla (17-day-old) exposed to UV-B radiation (145?µW?cm^?2 at the water surface) for three different exposure times: 5, 10 and 15?min was conducted. Fish without UV-B exposure served as control. UV-B radiation damaged both gill filaments and lamellae. The intensity of damage was minimal in 5?min exposed fish, followed by 10?min exposed fish and maximal in 15?min. The gill epithelium was severely damaged in 15?min irradiated fish compared to control. Pavement cells (PVCs) were damaged and the numbers of microridges within PVCs decreased. The deep boundary of PVC was not clear. In some area of gill epithelium, PVCs were destroyed and mitochondrion-rich cells (MRCs) were exposed. The 5?min exposure reduced the number of microridges in the PVCs, but the boundary of PVCs was still visible. MRCs in the gill epithelium were not exposed in 5?min exposed fish. The damage to PVCs and subsequent exposure of MRCs in UV-B irradiated fish may hamper respiratory functions and disturb osmoregulation in catla.     

Toxicity of leaf extract of yellow oleander Thevetia nerifolia on Tilapia

The usefulness of leaf extract as an ideal source of 'piscicide' in shrimp farming is described Leaf toxins are safe, eco-friendly and biodegradable. The fish Tilapia, Oreochromis mossambicus was exposed to the extract and the percentage of mortality at the end of 24,48, 72 and 96 hrs was recorded. The respective toxic range of aqueous, alcohol and acetone extracts of 24 hr LC50 and 96 hr LC50 values were found to be respectively 1118.79-330.30, 699.24-129.02 and 749.95-347.23 mg dry extract/litre for Oreochromis mossambicus. The LC50 values observed in different time periods in the fish exposed to aqueous extract were relatively high compared to the values obtained in acetone and alcohol extracts. Use of leaf extract, as piscicide in aquaculture farms is considered advantageous when viewed against the backdrop of using persistent chemicals.     

杀虫剂三唑磷对沼水蛙（Hylarana guentheri）蝌蚪的遗传毒性研究

采用微核试验和单细胞凝胶电泳技术(彗星试验)检测了不同浓度(0、0.2、0.4、1.2、2.0、2.8mg·L-1)的三唑磷溶液对沼水蛙蝌蚪遗传毒性的影响.结果表明,当三唑磷染毒24h(≥1.2mg·L-1)及72h(≥0.2mg·L-1)后,蝌蚪微核率和核异常率比对照组均有极显著的提高.在同一时间下,随着三唑磷浓度的增大,蝌蚪微核率和核异常率总体上随之增加;在同一浓度下,随着染毒时间的增加,低浓度组(≤0.4mg·L-1)微核率出现逐渐升高趋势;高浓度组(≥1.2mg·L-1)微核率则先增加后降低.染毒24h后,彗星试验的各项指标Tail Length、Comet Length、Tail Moment和Olive Tail Moment均随三唑磷浓度的增加而增加,并呈现极显著的剂量-效应关系.当三唑磷浓度达到0.2mg·L-1时,与对照组相比,DNA总体损伤水平(Comet Assay Tail Factor)显著提高.实验表明三唑磷对蛙类蝌蚪会造成一定的遗传损伤,微核试验和彗星试验在检测污染物对蝌蚪的遗传毒性方面具有较大的应用价值.     

Genotoxicity biomarkers in the amphipod Gammarus elvirae exposed in vivo to mercury and lead,and basal levels of DNA damage in two cell types

The present work deals with the application of genotoxicity biomarkers by means of the Comet assay in haemocytes and spermatozoa of the crustacean Gammarus elvirae exposed in vivo to heavy metals. Furthermore, a basal levels (BLs) study of DNA damage in the two cell types considered for two different gammarids species, G. elvirae and Echinogammarus veneris, was carried out. It is important to identify factors that influence the outcome of the assay in order to obtain reliable and reproducible results usable for risk assessment purposes. Our results highlight that the Italian legal limits for Hg and Pb, respectively, 0.5 and 50?µg/L, are inadequate for establishing safety thresholds in the aquatic environment. Furthermore, the freshwater invertebrate G. elvirae, used for the first time to measure the effect of genotoxicants, is a good candidate for evaluating the genotoxicity damage induced by heavy metals. Our results concerning spermatozoa show excessively variable responses and high BLs.     

Evaluation of developmental toxicity and apoptotic induction of the aqueous extract of Millettia pachycarpa using zebrafish as model organism

Extensive and indiscriminate use of synthetic compounds and natural compounds obtained from plant sources have resulted in serious threats to the aquatic ecosystem and human health. Aqueous extract of the root of the plant, Milletia pachycarpa Benth, is currently used for killing fish in the state of Manipur, India. Moreover, this plant is also used as traditional medicine in this region. Although it is widely used in traditional medicine, there is limited information available regarding the adverse effects and mechanism underlying its toxicity. This study examined the effects of exposure to aqueous extract of M. pachycarpa (AEMP) on early embryonic development of zebrafish embryos and mechanisms underlying toxicity. Zebrafish embryos treated with different concentrations of the AEMP produced embryonic lethality and developmental defects. The 96-hr-LC₅₀ of AEMP was found to be 4.276 µg/mL. Further, multiple developmental abnormalities such as pericardial edema, yolk sac edema, spinal curvature, swim bladder deflation, decreased heart rate, and delayed hatching were also observed in a dose-dependent manner. Zebrafish embryo showing moderate-to-severe developmental defects following AEMP exposure cannot swim properly. Further, this study examined oxidative stress and apoptosis in embryos exposed to AEMP. Enhanced production of ROS and apoptosis was found in brain, trunk, and tail of zebrafish embryos treated with AEMP. Data suggest that oxidative stress and apoptosis are associated with AEMP-induced embryonic lethality and developmental toxicity in zebrafish embryos.     

Cytotoxic and proinflammatory response of RAW 264.7 cells to differentially fractionated fungal fragments

Although significant research has been carried out for studying the impacts of pure mycotoxins on human cell, little information is known about the direct effects of fungal submicron particles. In this report, murine macrophage RAW 264.7 cells were exposed to fungal extracts from Stachybotrys chartarum RTI 5802 and Aspergillus versicolor RTI 3843 as well as mixtures from both fungi with fragment sizes ranging from 0.25 to 1?µm. Cell viability was found to be unaffected by all extracts except fragments in the 0.5–1?µm size range from A. versicolor RTI 3843, which decreased cell viability by approximately 60%. IL-1β expression was generally upregulated following exposure to fungal extracts from either S. chartarum RTI 5802 or A. versicolor RTI 3843. However, when extracts from the mixture of both fungi were introduced, IL-1β expression was downregulated. IL-6 and IL-10 expression were also generally upregulated. In comparison to the findings in the literature about the effects of pure mycotoxins on RAW 264.7 cells, the results presented herein show different patterns on mRNA expression which may be a result of the complex composition of the extract in terms of both mycotoxins and other fungal materials in addition to the potential interplay of the different components.     

Assessment of DNA damage and cytotoxicity of palmatine on human skin epithelial carcinoma cells

The present investigation was carried out to examine the cytotoxic and genotoxic effects of a Tinospora cordifolia crude methanolic extract (palmatine) on human skin epithelial carcinoma cells (A431). T. cordifolia is one of the indispensable medicinal plants used in Ayurvedic medicine for treatment of various diseases and recommended for improving the immune system. Cytotoxicity and genotoxicity evaluation was carried out using A431 cells treated with different concentrations of palmatine. The duration of the treatment was 24 and 48 hr. A cellular proliferative capacity test showed that palmatine produced cytotoxicity in concentration- and time-dependent manner. Further, palmatine induced significant intracellular reactive oxygen species generation and elevated lipid peroxidation, as well as activities of catalase and superoxide dismutase. DNA fragmentation analysis using the comet assay showed that palmatine induced genotoxicity in a concentration- and time-dependent manner. Evidence indicates palmatine is capable of induction of oxidative stress resulting in cell death and genomic instability.