Toxicity of tributyltin in the marine mollusc Mytilus edulis

Our previous studies have demonstrated that tributyltin (TBT) is genotoxic to the early life stages of marine mussels and worms. Here, the toxicity of TBT to adult organisms was determined using a suite of biomarkers designed to detect cytotoxic, immunotoxic and genotoxic effects. Exposure of adult mussels, Mytilus edulis, to environmentally realistic concentrations of TBTO for 7 days resulted in a statistically significant decrease in cell viability at concentrations of 0.5 μg/l and above. TBT had no effect on phagocytic activity or antioxidant capacity (FRAP assay). There was a statistically significant increase in DNA damage detected using the comet and micronucleus assays between the controls and 0.5, 1 and 5 μg/l of TBTO (P > 0.0005). Furthermore there was a strong correlation between DNA strand breaks (comet assay) and formation of micronuclei (P = 0.0005; R² = 61.5%). Possible mechanisms by which TBT could damage DNA either directly or indirectly are discussed including the possibility that TBT is genotoxic due to its ability to disrupt calcium homeostasis.     

Evaluating the genotoxic damage and hepatic tissue alterations in demersal fish species: a case study in the Ligurian Sea (NW-Mediterranean)

A protocol for detecting hepatic micronuclei in fish was performed to check genotoxic damage, as an indicator of environmental hydrocarbons exposure, in relation to the "Haven" oil spill. As target fish, we have chosen three demersal species with different habitats and feeding behaviour (i.e., Lepidorhombus boscii, Merluccius merluccius and Mullus barbatus) collected from two differently impacted areas and a control site. Additional analysis was performed by histological detection of hepatic tissue damages such as the presence of necrotic and tumour-like aspects. The three studied species showed different sensitivity to environmental pollutants exposure, L. boscii resulting the more sensitive in terms of both micronuclei incidence and tissue damage. The results of this study show that: (1) the micronucleus test could be an effective and fast method to detect oil pollution; (2) a clear response of L. boscii only to oil contamination for both micronucleus test and liver tissue alterations.     

Assessing surface water quality of the Yangtze Estuary with genotoxicity data

The present study investigated the genotoxicity of the surface water samples from the Yangtze Estuary with the Ames test in three seasons. Several important chemical parameters, such as COD inorganic nitrogen, active phosphate and heavy metals, were also analyzed at the same time. According the results, surface water samples from the south branch and a few of samples at the seaward end of the Yangtze Estuary show positive genotoxicity in some seasons. Chemical analysis revealed that the Yangtze Estuary was seriously polluted by inorganic nitrogen and by active phosphate. However, chemical parameters could not demonstrate the spatial variation of water quality of the estuary, and they could not assess adverse effects of chemicals in mixtures as well. Therefore, it is recommended that genotoxicity data, mutation rate of the Salmonella typhimurium strain TA98 at the dose of 1 l per plate, be taken as a new parameter assessing surface water quality of the Yangtze Estuary. Our studies also suggested that genetic toxicology assays, such as the Ames test, could be applied as a routine measure to monitoring marine environment in China. The paper proposes a development of the National Seawater Quality Standard of China.     

A statistical evaluation of the potential genotoxic activity in the surface waters of the Golden Horn Estuary

The potential genotoxic activity in the surface waters of the Golden Horn Estuary was statistically evaluated utilizing a combination of appropriate parametric and non-parametric tests. The genotoxic activities that were associated with the water samples were determined by the SOS chromotest microplate assay. This assay utilizes β-galactosidase activity, alkaline phosphatase activity, and four different solvent controls, to generate reliable results when corrected induction factors (CIF) are used as quantitative measurements of genotoxic activity. The CIF values were obtained from a total of 384 different genotoxic experiments that were grouped into subsets according to the respective seasons and the selected sampling locations. A total of 160 subsets were statistically compared to assess any possible differences between the pairs of groups, with 95% confidence limits. The findings of this study clearly indicate that some seasonal variations exist in the CIF values at several sampling sites. However, no potentially hazardous impact to the aquatic environment was found in the estuarine system.     

An evaluation of the health status of the Lavaca Bay, Texas ecosystem using Crassostrea virginica as the sentinel species

Locational risks for compromised ecosystem health for the eastern oysters (Crassostrea virginica) harvested from Lavaca Bay, Texas were estimated. Flow cytometric evaluation of variations in DNA content and the lysosomal destabilization assay were used for evaluation of genotoxicity and stress, respectively. Bayesian geo-statistical methods were utilized to estimate and evaluate spatial effects. For models with spatial risks, continuous surface maps of predicted parameter values were created to evaluate risk location. Lysosomal destabilization assay results were spatially oriented whereas flow cytometry results were fit best with the random effects model. While not spatially oriented, the highest levels of variations in DNA content were also present near industrial facilities. Locational risks of increased biomarkers of genotoxicity and stress in the eastern oyster (Crassostrea virginica) were increased with proximity to industrial facilities     

Using DNA damage to monitor water environment

DNA damage of aquatic organisms living in polluted environments can be used as a biomarker of the genotoxicity of toxic agents to organisms. This technique has been playing an important role in ecotoxicologlcal study and environmental risk assessment. In this article, main types of DNA damage caused by pollutants in water environments were reviewed; methods of detecting DNA damage were also documented for water environmental monitoring.     

Evidence of chromosomal damage in common eiders (Somateria mollissima) from the Baltic Sea

Common eiders nesting in the Baltic Sea are exposed to generally high levels of contaminants including potentially genotoxic polycyclic aromatic hydrocarbons and organochlorines. Blood samples were collected from eiders at eight sites in the Baltic Sea and two sites in the Beaufort Sea. DNA content variation was estimated using the flow cytometric method, and subsequently utilized as a biomarker of genetic damage. We observed no significant differences in genetic damage among populations within either the Baltic or Beaufort Seas. However, eider populations from the Baltic Sea had significantly elevated estimates of genetic damage compared to populations from the Beaufort Sea.     

Assessment of Cyto‐ and Genotoxic Effects of a Variety of Chemicals Using Saccharomyces cerevisiae

Saccharomyces cerevisiae as the most simple eukaryotic organism is broadly accepted as a laboratory model organism. For the detection of potential toxic effects of pure compounds and complex composed samples like wastewater a miniaturised short‐term in vitro cyto‐ and genotoxicity screening assay was developed. The assay based on genetically modified S. cerevisiae cells deleted in the prominent drug efflux transporters Pdr5, Snq2, and Yor1 that facilitate pleiotropic drug resistance. The yeast strain devoid of these proteins that mediate the efflux of structurally diverse hydrophobic compounds exhibited an increased sensitivity to a variety of organic compounds. The DNA damage inducible RAD54 promoter fused to a yeast optimized derivative of the GFP (green fluorescent protein) gene from the jelly fish Aequorea victoria served as an indicator of DNA damage in this strain. Various pure compounds including the direct‐acting genotoxins methyl‐N‐nitro‐N‐nitrosoguanidine (MNNG), 4‐nitroquinoline‐N‐oxide (4‐NQO), methyl methanesulfonate (MMS) and hydroxyurea as well as the heavy metals cadmium and chromium(VI), the insecticide lindane and the central nervous system stimulant caffeine were tested exhibiting dose dependent induction of green fluorescence. All compounds were in parallel examined for chronic toxicity. A bioassay detecting simultaneously geno‐ and cytotoxic effects of potential toxicants in a single assay can be an important tool with a variety of applications in environmental monitoring and aquatic ecotoxicology. By partial automation and miniaturisation to microtitration scale this bioassay enables sensitive and fast biomonitoring for a multitude of samples.     

Significance of an In Vivo Genotoxicity Test for Surface Waters Is Increased when the Physiological Characteristics of the Test Organism (Bivalvia: Mollusca) Are Taken into Account?

The alkaline filter elution assay using the gills of the freshwater clam species Corbicula fluminea detects breaks in single‐stranded DNA and is thus a good method for determining the genotoxic potential of surface waters. In attempting to standardize the procedure, a wide range of factors which could have an influence on the uptake of genotoxic substances by the exposed clams were studied. The most important parameters of the static exposure in relation to the rate of filtration by the animals turned out to be the temperature, the volume of the water, and the exposure time. Differences in body size and in the amount of suspended particles in surface waters did not play a significant role. The results demonstrate that the in vivo test system can be quite sensitive and its results reproducible when the relevant species‐specific characteristics of the test organisms are brought into consideration, even if the test organism belongs to a biologically more advanced group. A clear dose‐response relationship to the reference substances 4‐nitroquinoline‐1‐oxide (NQO) and N‐methyl‐N‐nitro‐N‐nitrosoguanidine (MNNG) can be observed even after a short exposure time of between four and twenty hours. No genotoxic effects were observed when using this method on surface waters from the area of Cologne (including water from the Rhine River and within the protection zone 2 of the Cologne waterworks).     

Comet assay for the detection of genotoxicity in blood cells of flounder (Paralichthys olivaceus) exposed to sediments and polycyclic aromatic hydrocarbons

To investigate the genotoxic effect of marine sediments on aquatic organism, sediment samples were collected from 13 sites along the coast of Gwangyang Bay (Korea). Concentrations of polycyclic aromatic hydrocarbons (PAHs) in sediments were determined and the relationship between exposure of flounder blood cells to sediment extracts and DNA single-strand breakage in the blood cells was examined using the comet assay. Levels of DNA damage were proportionally increased by exposure concentration and the highest sediment-associated DNA damage was observed at the station showing the highest PAHs contamination. DNA damage in blood cells exposed to five types of PAHs (benzo[a]pyrene, fluoranthene, anthracene, pyrene and phenanthrene) and in flounder (Paralichthys olivaceus) exposed to benzo[a]pyrene (BaP) for 0, 2 and 4 days were assessed by measuring comet tail length. The tail lengths of five PAHs-exposed groups at 50 and 100 ppb were significantly different from the non-exposed group, and the genotoxic effect of BaP correlated with both concentration and duration of exposure. Throughout the study, significant differences in DNA breakage were recorded between cells exposed to sediment extracts or PAHs and non-exposed control. This study demonstrated the comet assay as a successful tool in monitoring contamination of marine sediments and assessing genotoxicity of PAHs in marine organisms, either in vitro or in vivo.