THE NAN YE LI SANMENIAN FOSSILIFEROUS DEPOSIT

Nan Ye Li is a little village, lying about 80 kilometers West from Cheng Ting Fu, along the mountains, at some 370 meters above the level of the great Hopei plain.     

Time-course evaluation of ROS-mediated toxicity in mussels, Mytilus galloprovincialis, during a field translocation experiment

Harbours can be considered as model environments for developing and validating field monitoring procedures and to investigate mechanistic relationships between different biological responses. In this study several biomarkers were investigated in marine mussels caged for 4 weeks (June-July 2001) into an industrialized harbour of NW Italy. Organisms were collected at different time-intervals to better characterize the sensitivity, temporal variations and interactions of analysed responses. Besides single antioxidants the total oxyradical scavenging capacity (TOSC) assay was used to analyse the capability of the whole antioxidant system to neutralize specific forms of radicals: these data were further integrated by measurement of DNA integrity, oxidized bases and the impairment of lysosomal membrane stability in haemocytes. Results showed a biphasic trend for single antioxidants and TOSC, with an increase during the first 2 weeks of exposure to the polluted site followed by a progressive decrease up to a severe depletion in the final part of the experiment.     

Oxidative damage to DNA: an immunohistochemical approach for detection of 7,8-dihydro-8-oxodeoxyguanosine in marine organisms

The modified nucleoside 7,8-dihydro-8-oxodeoxyguanosine (8-oxo-dG) is an index of oxidative DNA damage. An immunohistochemical approach based on the use of monoclonal antibody 1F7 against 8-oxo-dG was investigated in marine organisms with immunoperoxidase and immunofluorescent detection. Relative staining intensity as a measure of the 8-oxo-dG level was microscopically assessed. After laboratory exposures to benzo[a]pyrene (B[a]P), higher levels of oxidative DNA damage were clearly detected in all treated animals compared to controls. While this method eliminates DNA extraction reducing the processing of biological samples, absolute values are not provided. Further, the method requires only small amounts of tissue and potentially discriminates susceptibility to oxidative damage in different cell types. These results suggest that the assay should have practical applications in marine ecotoxicology.     

Effect of intertidal compared to subtidal exposure on the uptake,loss and oxidative toxicity of water-born benzo[a]pyrene in the mantle and whole tissues of the mussel,Mytilus edulis L

Blue mussels (Mytilus edulis, L.) were exposed to a single dose of 1 ppb benzo[a]pyrene (BaP) under subtidal (SC) or tidal conditions (TC; 6 h immersion, 6 h emersion) in order to follow its bioaccumulation in whole mussel and mantle tissue, and to compare BaP-mediated toxicity on lipids (malonaldehyde formation, MDA) in the mantle. Rapid uptake of BaP (70-80% of BaP initially introduced in tanks) was observed in both conditions after 12 h, but subsequent depuration in clean water was slower in TC mussels. BaP levels decreased in whole tissue in both conditions between 12 and 24 h, but increased in mantle. The mantle BaP levels were similar during the first 4 days in SC and TC, but whereas they decreased in SC after 7 days. BaP was retained at high levels in mantle in TC until the end of the study (14 days). In both conditions, significant increases (P < 0.05) in lipid peroxidation were observed after 4 days, but MDA levels were approximately 3 times higher in the mantle of TC than SC mussels, although BaP tissue concentrations were similar. These observations suggested that increased BaP-mediated toxicity in mantle lipid was due to the interactive effect of the tidal cycle of immersion/emersion on BaP-mediated oxidative damage.     

Antioxidant efficiency and detoxification enzymes in spotted dogfish Scyliorhinus canicula

Although elasmobranchs are widely distributed species, commonly found on sandy, gravely or muddy bottoms, several biological aspects of their metabolism still remain poorly investigated. In this work the efficiency of antioxidant system and detoxification enzymes were investigated in the coastal spotted dogfish Scyliorhinus canicula and in the red mullet Mullus barbatus for comparison with a teleost species. Organisms were sampled during a bottom trawl survey and analyzed for the biotransformation activity (EROD), levels of metallothioneins, catalase, glutathione S-transferases and total oxyradical scavenging capacity (TOSC) toward peroxyl radicals and hydroxyl radicals. EROD activity in the elasmobranchs was more than one order of magnitude lower than in the red mullets, while similar levels of metallothioneins were measured in these species. S. canicula showed significantly lower antioxidant enzymes and a more reduced efficiency in neutralizing *OH; on the other hand the scavenging capability toward ROO* was comparable in S. canicula and M. barbatus.     

Total oxyradical scavenging capacity and cell membrane stability of haemocytes of the Arctic scallop,Chlamys islandicus,following benzo(a)pyrene exposure

Industrial activities, notably oil and gas industries, are expanding in the Arctic. Most of the biomarkers were developed using temperate organisms living at temperatures above 10 degrees C. Little is known about the biomarker responses of organisms living between -1.88 and 5 degrees C. Therefore, assessment of the toxicity of chemicals to cold-water adapted species is required. In this study, the Arctic scallop, Chlamys islandicus, was selected as a key species for bio-monitoring because of wide distribution in Arctic waters and its commercial value. Test animals, stored in seawater at 2 degrees C, were injected with benzo(a)pyrene (diluted in cod liver oil 5 mg ml(-1)) in the adductor muscle every 24 h for four days giving a final dose of 0, 74 and 90.6 mg kg(-1) wet weight for control, low and high dose, respectively. The biomarkers used were total oxyradical scavenging capacity (TOSC) in the digestive gland and cell membrane stability of haemocytes. TOSC values were significantly reduced (ca. 30%) in exposed groups (P < 0.05), indicating a depletion in oxyradical molecular scavengers. The antioxidant defences appeared to be overwhelmed by the reactive oxygen species as the plasma membranes of haemocytes were destabilised (P < 0.05) probably due to lipid peroxidation. These data indicate that reactive oxygen species (ROS) were produced by Arctic scallops via the metabolisation of benzo(a)pyrene at 2 degrees C.     

Cell responses to xenobiotics: comparison of MCF7 multi-drug- and mussel blood cell multi-xenobiotic-defense mechanisms

Multi-drug resistance (MDR) in MCF7 breast cancer cells and multi-xenobiotic resistance (MXR) in mussel (Mytilus edulis) blood cells (MBC) are well known mechanisms that contribute to the decrease in intracellular concentrations of many unrelated but cytotoxic compounds. In the present work, we have carried out comparative investigations of the MDR/MXR protective mechanisms using a rapid colorimetric assay for cell viability and calcein accumulation for MDR/MXR activities. These studies were performed using cultured MCF7 and MBC before and after in vitro exposure to xenobiotics. Our results indicate that a 5-day exposure to doxorubicin or vincristine decreased calcein accumulation in MBC which is consistent with an induction of multi-xenobiotic resistance. The increase in calcein accumulation provoked by 1-h treatment with 50 microM verapamil was much lower in MBC when compared to the P-glycoprotein overexpressing MCF7 cell line. We conclude that such microplate assays could be used in primary cultures of MBC to estimate the effects of various chemicals on MXR activity.