Peter-Diedrich Hansen

Cholinesterases of marine teleost fish: enzymological characterization and potential use in the monitoring of neurotoxic contamination

Abstract Acetylcholinesterase (AChE) is well established as a biomarker of exposure to organophosphate compounds in freshwater fish. By constrast, only a few studies on the enzymology and responsiveness of AChE are available for marine species. In this study, we report characteristics of cholinesterases from brain and muscle tissue of three marine teleosts, Limanda limanda, Platichthys flesus and Serranus cabrilla, to provide basal information for environmental monitoring in coastal and marine areas. In brain, cholinesterase activity was exclusively comprised of AChE. In contrast, both butyrylcholinesterase (BChE) and AChE were present in axial muscle. The fish BChEs displayed atypical substrate and inhibitor specificities in that they cleaved acetyl-β-(methyl)thiocholine and were sensitive to BW284C51. Bimolecular inhibition constants, Ki, of BChE for the organophosphates paraoxon and dichlorvos were more than three orders of magnitude greater than the respective Ki values of AChEs. This reflects a much higher sensitivity of BChE to the organophosphates than AChE. Neither AChE from muscle and brain within one species, respectively, nor corresponding cholinesterases from different species differed significantly in their Ki values. The presence of two cholinesterases of differential sensitivity in the same tissue is potentially useful in monitoring.

Bioaccumulating substances and reproductive success in baltic flounder platichthys flesus

Abstract Running ripe Baltic flounder Piatichthys flesus were caught with a bottom trawli. Eggs were stripped and artificially inseminated and incubated in Baltic sea water at 6.8 °C and a salinity of 27.5% After hatching, siraight and viable appearing larvae — viable hatch — were determined. Ovary and liver samples of the female parental fish were collected and analyzed for chlorinated hydrocarbons and heavy metals. Viable hatch was significantly affected at ovary chlorinated hiphenyl (PCB) levels higher than 120 ng/g wet weight. Thus 120 ng/g PCB was designated a threshold level in flounder eggs. above which reduced survival of developing eggs and larvas can be expected. Other chlorinated hydrocaihons. and metals did not show any correlation between ovary and/or liver contamination levels and viable hatch. The melhod employed is deemed to be a promising tool for the assessment of pollution effects on marine biola.

The cytotoxic and genotoxic potential of surface water and wastewater effluents as determined by bioluminescence, umu-assays and selected biomarkers

Two bacterial tests employing Photobacterieum phosphoreum (Microtox bioluminescence test) and Salmonella typhimurium TA 1535 pSK1002 (umu-assay) were evaluated to estimate the cytotoxic and genotoxic potential of water samples from the selected rivers in Germany as well as the primary and secondary effluents of some sewage treatment plants. Rainbow trout (Onchorynchus mykiss) were exposed to different concentrations (20-40%) of secondary effluent in the model online aquatic monitoring plant WaBoLu-Aquatox. The toxic potential of water samples from the exposure tanks was determined in two prokaryotic test systems and the biomarkers acethylcholinesterase (AChE) activity in muscle tissue and DNA unwinding assay in liver tissue of fish. Samples from the tested rivers showed no inhibition of the bioluminescence of P. phosphoreum or growth of umu-bacteria. Only primary effluent samples from the treatment plants at the Saale River inhibited the light emission or the growth of test bacteria by more than 20%. The induction ratio of umu-bacteria was in most of the river samples less than the threshold for genotoxicity (IR < 1.5). Only some samples from the Saale River, especially at sites downstream of secondary effluents caused genotoxic responses in the umu-assay. Samples of primary effluents contained the greatest genotoxic potential up to GEUI = 6 which was not detectable in samples of secondary effluents. A concentration range 20-40% secondary effluent inhibited AChE activity in muscle tissue and significantly increased DNA fragmentation in liver tissue of rainbow trout. In contrast, no cytotoxic or genotoxic responses in the umu-assay were caused by water samples. Both bacterial methods can be successfully used to analyse the cytotoxic and genotoxic response of industrial and domestic wastewater and to estimate the effectiveness of sewage treatment units. However, because of their low sensitivity and high susceptibility, they are not reliable as a single test for the detection of cytotoxicity and genotoxicity in surface water. The application of prokaryotic tests systems with biomarkers such as AChE activity and DNA fragmentation in different tissues of test organisms seems to be a useful combination for the assessment of cytotoxic and genotoxic potential in surface water and secondary effluent.

Chlorinated hydrocarbons and hatching success in Baltic herring spring spawners

Abstract Eggs from 69 females of spring spawning herring from the German Baltic coast (Travemunde, April 1979) were incubated in clean sea water (20‰ S, temperature 8°C) under standard conditions. Sixty-one trials could be used for the evaluation of hatching success. Viable hatch was taken as a measure to evaluate the effects of chlorinated hydrocarbons accumulated in gonads, liver and muscle of parental fish. PCB levels in running ripe females ranged on a wet weight basis between 19 and 241 ng g −1 (gonad), 20 and 377 ng g −1 (liver) and 11 and 1820 ng g −1 (muscle). Concentrations of other chlorinated hydrocarbons (DDD, DDE, γ-HCH, etc.) were in the same range as reported by other authors for Baltic herring (Huschenbeth, 1973, 1977). Viable hatch was significantly affected at ovary DDE concentrations higher than 18 ng g −1 (wet wt) and PCB concentrations of more than 120 ng g −1 (wet wt). Results are compared with data obtained during earlier investigations with flounder eggs.

Marine molluscs and fish as biomarkers of pollution stress in littoral regions of the Red Sea, Mediterranean Sea and North Sea

Abstract The intensive development of industry and urban structures along the seashores of the world, as well as the immense increase in marine transportation and other activities, has resulted in the deposition of thousands of new chemicals and organic compounds, endangering the existence of organisms and ecosystems. The conventional single biomarker methods used in ecological assessment studies cannot provide an adequate base for environmental health assessment, management and sustainability planning. The present study uses a set of novel biochemical, physiological, cytogenetic and morphological methods to characterize the state of health of selected molluscs and fish along the shores of the German North Sea, as well as the Israeli Mediterranean and Red Sea. The methods include measurement of activity of multixenobiotic resistance-mediated transporter (MXRtr) and the system of active transport of organic anions (SATOA) as indicators of antixenobiotic defence; glutathione S-transferase (GST) activity as an indicator of biotransformation of xenobiotics; DNA unwinding as a marker of genotoxicity; micronucleus test for clastogenicity; levels of phagocytosis for immunotoxicity; cholinesterase (ChE) activity and level of catecholamines as indicators of neurotoxicity; permeability of external epithelia to anionic hydrophilic probe, intralysosomal accumulation of cationic amphiphilic probe and activity of non-specific esterases as indicators of cell/tissue viability. Complete histopathological examination was used for diagnostics of environmental pathology. The obtained data show that the activity of the defensive pumps, MXRtr and SATOA in the studied organisms was significantly higher in the surface epithelia of molluscs from a polluted site than that of the same species from control, unpolluted stations, providing clear evidence of response to stress. Enhanced frequency of DNA lesions (alkaline and acidic DNA unwinding) and micronucleus-containing cells was significantly higher in samples from polluted sites in comparison to those from the clean sites that exhibited genotoxic and clastogenic activity of the pollutants. In all the studied molluscs a negative correlation was found between the MXRtr levels of activity and the frequency of micronucleus-containing hemocytes. The expression of this was in accordance with the level of pollution. The complete histopathological examination demonstrates significantly higher frequencies of pathological alterations in organs of animals from polluted sites. A strong negative correlation was found between the frequency of these alterations and MXRtr activity in the same specimens. In addition to these parameters, a decrease in the viability was noted in molluscs from the polluted sites, but ChE activities remained similar at most sites. The methods applied in our study unmasked numerous early cryptic responses and negative alterations of health in populations of marine biota sampled from the polluted sites. This demonstrates that genotoxic, clastogenic and pathogenic xenobiotics are present and act in the studied sites and this knowledge can provide a reliable base for consideration for sustainable development.

Toxicity of domoic acid in the marine mussel Mytilus edulis.

The neurotoxic, immunotoxic and genotoxic effects of domoic acid (DA) on the blue mussel Mytilus edulis were investigated by biomarkers, acethylcholinesterase (ChE) activity in gills, DNA fragmentation in digestive glands, vitality and phagocytosis activity of haemocytes in haemolymph of mussels. After intra muscular injection of DA at the concentrations ranging from 1-500 ng/g body weight (bw), no neurotoxic effect was detected within incubation times of 48 h and 7 d. The vitality of haemocytes remained in all mussels at the level of control samples within 48 h, and increased significantly after 7 d (P<0.05). At DA concentrations ranging from 1 to 100 ng/g bw haemocytes suggested a great phagocytosis activity, but no alteration in their number by both incubation times. By increasing DA concentration of 500 ng/g bw, the number of haemocytes doubled in 48 h without any change in phagocytosis activity. Primary DNA lesions in digestive glands of all injected mussels were determined in acute phase of poisoning within 48 h, and rapidly repaired after 7 d of incubation.

Estrogenic potential of halogenated derivatives of nonylphenol ethoxylates and carboxylates

Halogenated derivatives of nonylphenol and of its alkylates are generated during drinking water disinfection and treatment procedures. In this paper we analyze the potential of these compounds to interact with the estrogen receptor and to activate hormone-regulated gene promoters. We used the recombinant yeast assay (RYA) and the human breast cancer cell MCF7 proliferation assay for both estrogenic and antiestrogenic activities and the enzyme-linked receptor assay to examine in vitro binding to the receptor. Many nonylphenol derivatives were very weak estrogens in our functional tests when compared to nonylphenol while retaining a substantial affinity for the estrogen receptor in vitro. Antiestrogenicity tests demonstrated that brominated nonylphenol and most of the carboxylated compounds studied here behaved as estrogenic antagonists in the RYA. We also detected an increased cytotoxicity for the carboxylated derivatives in both yeast and mammalian cells. We conclude that derivatization may mask the apparent estrogenicity of nonylphenol, but the resulting compounds still represent a potential hazard since they are still able to bind the estrogen receptor and to influence the physiological response to estrogens. Our results also illustrate the advantage of combining different methods to assay estrogenicity of unknown substances.

Parasites of flounder ( Platichthys flesus L.) from the German Bight, North Sea, and their potential use in biological effects monitoring

In the frame of an integrated biological effect monitoring programme, the parasite community of flounder (Platichthys flesus) was investigated at different locations in the German Bight from 1995 to 2000. In order to assess the impact of environmental contamination caused by anthropogenic activities on the parasite community, selected parasitological parameters that displayed significant differences between the sampling sites were subjected to correlation analyses with site-specific contamination and individual pollution loads of their fish hosts. In addition, correlation analyses were conducted with the responses of selected genetic, biochemical, histopathological, physiological and immunological parameters of fish, used as potential biomarkers. In total, 802 flounder were analysed for these parameters. Information on the chemical background at the sampling sites was derived from sediment samples and from 120 samples of blue mussel (Mytilus edulis) tissue, collected at each of the sampling sites. Based on chemical data available from the sediment and blue mussel samples, a pollution gradient could be established between the sampling sites for individual contaminants. The relative abundance of Acanthochondria cornuta, Cucullanus heterochrous and Zoogonoides viviparus, and the community measures species richness and number of heteroxenous species decreased with increasing concentrations of individual heavy metals or hydrocarbons in sediment and blue mussel samples. Most of the parasitological parameters significantly reflected the established site-specific contamination gradient, when data were pooled over all sampling campaigns. Significant correlations were also found with the contamination level of individual flounder. The parasitological parameters included the parasite species Lepeophtheirus pectoralis and Lernaeocera branchialis, which were not correlated to site-specific contamination. Several biomarkers were significantly correlated to the abundance of parasitic copepods A. cornuta, Lernaeocera branchialis and Lepeophtheirus pectoralis and to parasite community parameters. The results showed that the abundance of several metazoan parasite species, species richness and parasite diversity were reduced in contaminated habitats, and that differences between sites were not only related to natural factors, such as salinity, but also to pollution-induced stress. Thus, it can be concluded that the parasite community of fish responds to the level of pollution at a specific site as well as to residues of xenobiotics in individual fish. These findings give indications that the parasite community of fish is a valuable parameter for the assessment of ecological consequences of chemical contamination in aquatic habitats.

Multivariate statistical approach to the temporal and spatial patterns of selected bioindicators observed in the North Sea during the years 1995–1997

Abstract A comprehensive database, containing biological and chemical information, collected in the framework of the bilateral interdisciplinary MARS project (”biological indicators of natural and man-made changes in marine and coastal waters”) during the years 1995–1997 in the coastal environment of the North Sea, was subjected to a multivariate statistical evaluation. The MARS project was designated to combine a variety of approaches and to develop a set of methods for the employment of biological indicators in pollution monitoring and environmental quality assessment. In total, nine ship cruises to four coastal sampling sites were conducted; 765 fish and 384 mussel samples were analysed for biological and chemical parameters. Additional information on the chemical background at the sampling sites was derived from sediment samples, collected at each of the four sampling sites. Based on the available chemical data in sediments and black mussel (Mytilus edulis) a pollution gradient between the selected sites, was established. The chemical body burden of flounder (Platichthys flesus) from these sites, though, did not reflect this gradient equally clear. In contrast, the biological information derived from measurements in fish samples displayed significant a regional as well as a temporal pattern. A multivariate bioindicator data matrix was evaluated employing a factor analysis model to identify relations between selected biological indicators, and to improve the understanding of a regional and temporal component in the parameter response. In a second approach, applying the k-means algorithm on the data matrix, two significantly different clusters of samples, characterised by the current health status of the fish, were extracted. Using this classification a temporal, and in the second order, a less pronounced spatial effect was evident. In particular, during July 1996, a clear sign of deteriorating environmental conditions was extracted from the biological data matrix.

Biosensors for environmental monitoring of aquatic systems

Biosensors for environmental monitoring of aquatic systems , Biosensors for environmental monitoring of aquatic systems , کتابخانه دیجیتال جندی شاپور اهواز