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The paper describes the technical, procedural and biological aspects of 7-ethoxyresorufin O-deethylase (EROD) activity measurements in dab, Limanda limanda, from the North Sea. Several sources of measurement error in assays of EROD activity measurements for biological effects monitoring exercises could be defined, particularly for assay conditions, sample storage and sampling. In addition to variability caused by technical errors, biological features inherent in the surveyed fish may lead to even more severe errors in EROD activity measurements. This is particularly true of the choice of sampling time. Disregarding the species-specific spawning periods may lead to a loss of activity by a factor of as much as 90 in females and 40 in males, thus making it necessary to design sampling surveys appropriately. [ABSTRACT FROM AUTHOR]

Ethoxyresorufin O-deethylase (EROD) activity was measured in the liver of dab ( Limanda limanda) and flounder ( Platichthys flesus) from the German Bight (southern North Sea) and compared with muscle and liver polychlorinated biphenyl (PCB) concentrations in an attempt to relate EROD activity to PCB body burden. In none of the different datasets (species-, tissue- or matrix-dependent) was a significant ( P<0.05) correlation between PCB tissue contamination and EROD activity found. Yet EROD activity was significantly correlated with polycyclic aromatic hydrocarbons (PAH) levels (phenanthrene, fluoranthene, pyrene) in muscle tissue, indicating a possible dependence of EROD expression on other ubiquitous organic contaminants, thus making it a suitable biomarker for general pollution. [ABSTRACT FROM AUTHOR]

An integrated biological effect monitoring concept has been tested in flounder ( Platichthys flesus L.) from four locations with different anthropogenic impact in the German Bight. During 3 years of sampling, biomarkers at all levels of biological organisation from the molecular to the ecosystem level were applied and tested on 742 individual fish of body lengths between 18 and 25 cm. At the ecosystem level, the fish were taken as a habitat for the parasite assemblage. The hypothesis was that changes in the environment might lead to changes in the species diversity of parasites and in the infection intensity of single species, as well as between heteroxenic and monoxenic parasite species (H/M ratio). At the molecular level, activity of the CYP1A-dependent monooxygenase ethoxyresorufin O-deethylase (EROD) was used as a biomarker of exposure. At the subcellular level, the integrity of lysosomal membranes in hepatocytes was taken as an indicator of non-specific acute and chronic toxic effects. Both biomarkers are recommended by the ICES Advisory Committee on the Marine Environment for the application in biological effects monitoring programmes. In addition, neutral lipid content in the liver was used as a marker for pathologically induced fat accumulation. In the same individual fish, a new method for the measurement of macrophage aggregate activity in the liver was tested for its application and reliability in reflecting immunosuppression. Tests were accompanied by chemical analysis of standard organochlorine and heavy metal residues in flounder tissue. A total of 33 parasite species were found. As an indicator species, the mean abundance of Trichodina sp. reflected best the pollution gradient observed with highest infection intensity at the most polluted location. Species diversity was significantly higher in fish caught near the reference site and significantly lower in fish from the polluted Elbe estuary. The use of the heteroxenous/monoxenous species ratio as a marker was not useful at the locations investigated because of the dominance of heteroxenous species at all habitats. Since EROD activity and macrophage aggregate activity were dependent on sex and maturity of female flounder, only male fish were taken into consideration for the integrated evaluation of data. All biochemical and histochemical tests were able to reflect accurately the site-specific differences, as well as an observed pollution event at the end of 1995 as determined by chemical analyses. The correlation analysis revealed a connection not only between the single parasitological and biochemical parameters but also within these groups. The non-specific immune response and Trichodina infection intensity were correlated with all other parameters, leading to the assumption that these may serve as links between the lowest and the highest levels of biological organisation. The simultaneous use of metabolic and parasitological results facilitated the interpretation of the observed variations of the data and the distinction between natural variation and pollution-mediated effects. [ABSTRACT FROM AUTHOR]

The complex interactions between parasites, hosts and the environment are influenced by the stability of the ecosystem. Heteroxenous parasites, with complex, multiple-host life cycles, can persist only in habitats where the full range of their required hosts are present. Conversely, in impoverished environments such as those impacted by environmental stress, monoxenous species that have simple, single-host life cycles are likely to predominate. In the present study, we analyzed the ratio between heteroxenous and monoxenous (H/M) parasites as well as parasite species richness (SH/SM) and species diversity in rabbitfish ( Siganus rivulatus) collected from several sites in the Red Sea. The rabbitfish is a Suez Canal immigrant, well established in the eastern Mediterranean, and fish were also collected from a site on the Mediterranean coast of Israel. Separate treatment of the micro- and macroparasite components of the rabbitfish parasite communities in the Red Sea suggested that macroparasites only – monogenea and gut parasites – were better indicators than the parasite community as a whole. Quantification of macroparasites is accurate, saves time and effort, produces more accurate data and better differentiates between sites. Higher H/M ratios and SH/SM ratios were found in the rabbitfish collected at the ecologically stable habitat of the coral reef compared to rabbitfish from sandy habitat or mariculture-impacted sandy habitat. The results of the study emphasized the negative impacts of cage mariculture on the environment. The rabbitfish collected near the mariculture farms supported the poorest and least diverse parasite communities of all sampled sites, with virtual depletion of heteroxenous species, and even reduction of gill monogenean infections on the hosts. When results from the Mediterranean sites were compared with those of the Red Sea, the data showed full representation of monoxenous parasites (all but one of Red Sea origin), while heteroxenous species were completely absent. We may therefore regard the Mediterranean as a simulation model for a severely environmentally deteriorated, impoverished habitat, in which all or part of the intermediate host species have been depleted, enabling survival of the monoxenous parasite species only. Parasitological investigations were supplemented by testing the activity of cytochrome P 450- dependent mono-oxygenase EROD as a measure of exposure, and lysosomal stability as a measure of toxic effect in the liver of rabbitfish. The results underline the parasitological findings, showing that fish caught at the impacted sandy beach location in the Red Sea have significantly higher EROD activity and a decreased membrane stability compared with animals from the coral reef. In comparison, EROD activity values in rabbitfish from the Mediterranean Sea were double, while lysosomal membrane stability was half that measured at the most impacted Red Sea location. [ABSTRACT FROM AUTHOR]

The "bioeffect assessment index" (BAI) is based on the integration of several pathological endpoints measured in the liver of European flounder (Platichthys flesus (L.)) during a long term study of biological effects of pollution in the German Bight. The BAI represents a modification of the "health assessment index" since it includes solely validated biomarkers reflecting toxically induced alterations at different levels of biological organisation in order to quantify the effects of environmental pollution. The concept of the BAI is based on the observation of progressive deleterious effects from early responses to late effects. Specific "key events" were detected, representing progressive stages of functional deterioration. The biomarkers selected from a whole battery of cellular markers for the BAI calculation reflect deleterious effects of various classes of contaminants such as heavy metals, organochlorines, pesticides, PAHs, and therefore reflect general toxicity in an integrative manner. Selected biomarkers were: lysosomal perturbations (reduced membrane stability), storage disorders (lipid accumulation) as early markers for toxic effects of liver cells, and the size of macrophage aggregates and their acid phosphatase activity. The latter two markers are indicative for the modulation of non-specific immune response which represents longer time scale responses after chronic exposure.

From 1995 to 2000 biological effects were studied in liver of flounder (Platichtysflesus L.) from the German Bight. During the study period deleterious consequences of acute discharges of DDT and PCBs in early spring 1996 and after 1998 due to remobilization of contaminants from riverbed deepening of the River Elbe became evident. As core biomarker which reflects toxically induced liver pathologies and integrates effects of various classes of pollutants we measured the integrity of lysosomal membranes in individual flounder liver. During the study period, twice statistically significant disturbances of lysosomal function was detected in fish from the River Elbe: in summer 1996 and in spring 1999. Yet, the detrimental contaminant effects were not only restricted to individuals from the Elbe but expanded to those flounder inhabiting formerly less polluted reference areas. In contrast to flounder of the Elbe, their ability to recover from the lysosomal disorders were limited. While in autumn 2000 Elbe individuals showed clear signs of recovery, those fish caught in areas more distant to the source of toxicant input still maintained significantly decreased lysosomal membrane integrity. It can be speculated that fish populations which are not continuously exposed to chronic anthropogenic stress may have a lower potential or need a longer period to recover from the effects of pollution.