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P J den Besten

Publications and source records attributed to P J den Besten.

10 recordsLinked to original sources

Bioaccumulation and biomarkers in the sea star Asterias rubens (Echinodermata: Asteroidea): a North Sea field study.

During two North Sea field trips in March 1995 and September 1996 sea stars, Asterias rubens, were collected at various stations along pollution gradients in order to study the relation between biochemical markers and levels of accumulated contaminants. Biomarkers measured were: cytochrome P450 level, benzo[a]pyrene hydroxylase (BPH) activity, acetylcholinesterase (AChE) activity and DNA integrity. Accumulation levels of heavy metals, polychlorinated biphenyls, organochlorine pesticides and polycyclic aromatic hydrocarbons (PAHs) in the pyloric caeca of sea stars indicate different pollution gradients, influenced by rivers in The Netherlands, UK and Germany. For some contaminants, especially PAHs, relatively high levels were found in the central part of the North Sea (Dogger Bank). On the basis of multivariate statistics, stations near the mouth of the Elbe and the Rhine/Meuse were shown to have different patterns of biomarker responses. Sea stars from stations in coastal zones showed relatively high levels of cytochrome P450 and 'P418', another haemoprotein that is present in most marine invertebrates. The station nearest to the Elbe Estuary showed the lowest BPH and AChE activity. DNA integrity was lower especially in stations near the Dutch coast and in a station near the Tees/Tyne estuaries. Using these biomarkers as early warning signals of exposure and/or adverse effects, this type of monitoring can be used also in the future to study the spatial and temporal trends in the quality of coastal waters.

Acetylcholinesterase↗

Evaluation of bioassays versus contaminant concentrations in explaining the macroinvertebrate community structure in the Rhine-Meuse delta, The Netherlands.

It is often assumed that bioassays are better descriptors of sediment toxicity than toxicant concentrations and that ecological factors are more important than toxicants in structuring macroinvertebrate communities. In the period 1992 to 1995, data were collected in the enclosed Rhine-Meuse delta, The Netherlands, on macroinvertebrates, sediment toxicity, sediment contaminant concentrations, and ecological factors. The effect of various groups of pollutants (polycyclic aromatic hydrocarbons, trace metals, oil, polychlorinated biphenyls) and of ecological variables on the structure of the macroinvertebrate community were quantified. Ecological factors explained 17.3% of the macroinvertebrate variation, while contaminants explained 13.8%. Another 14.7% was explained by the covariation between ecological variables and contaminants. Polycyclic aromatic hydrocarbons explained a larger part of the variation than trace metals. The contributions of oil and polychlorinated biphenyls were small but significant. Elevated contaminant concentrations were significantly associated with differences in the macroinvertebrate food web structure. The response in bioassays (Vibrio fischeri, Daphnia magna, Chironomus riparius) was susceptible to certain contaminants but also to certain ecological factors. There was a weak correlation between in situ species composition and bioassays; 1.9% of in situ macroinvertebrate variation was explained by the bioassay responses. This seems to contradict the validity of using bioassays for a system-oriented risk assessment. Possible reasons for this discrepancy might be the manipulations of the sediment before the test and a higher pollutant tolerance of the in situ macroinvertebrates. Thus, macroinvertebrate field surveys and laboratory bioassays yield different types of information on ecotoxicological effects, and both are recommended in sediment risk assessment procedures.

Animals↗

Cytochrome P450 monooxygenase system in echinoderms.

The results of a limited number of studies on echinoderms provide evidence for the presence of a cytochrome P450 monooxygenase system in representatives of three classes of the phylum Echinodermata: the asteroids (sea stars), holothuroids (sea cucumbers) and echinoids (sea urchins). The monooxygenase system has been demonstrated to be involved in the metabolism of xenobiotic compounds, but is assumed to have its primary function in the metabolism of endogenous substrates, such as steroids. Available data on P450 cofactor requirement, P450-dependent metabolism of benzo[a]pyrene, studies with classical inhibitors of P450, specificity of P450 induction by planar compounds, and the changes in the benzo[a]pyrene metabolite profile in induced animals suggest similarities with the MO system present in vertebrates. However, the relatively high capacity of the monooxygenase system in sea stars to catalyse reactions with organic hydroperoxide as donor for activated oxygen, and the low induceability during exposure to xenobiotics indicate also important differences between the echinoderm cytochrome P450 monooxygenase system and that of vertebrates. Some evidence was found for the existence of different forms of cytochrome P450 in sea stars. Catalytic functions of the cytochrome P450 monooxygenase system of sea stars in the metabolism of steroids may be suppressed as a result of the induction of cytochrome P450 by xenobiotics.

Animals↗

Molecular responses as indicators of marine pollution: DNA damage and enzyme induction in Limanda limanda and Asterias rubens.

During a survey from 26 August through 13 September 1991, specimens of the flatfish, Limanda limanda (dab), and the asteroid echinoderm Asterias rubens (seastar), were collected at sampling locations along transects radiating into the North Sea from the coastal zone of The Netherlands. In homogenates of liver tissue from male dab and the digestive gland (pyloric caeca) of female seastar, DNA damage (strand breaks) and induction of the cytochrome P450-dependent monooxygenase system (MO) were determined. Areas could be described with significantly increased percentages of strand breaks (lower integrity) both in dab and seastar. However, enhanced DNA strand breaks did not correspond with contamination gradients, expressed as concentrations of polychlorinated biphenyls (PCBs) or polyaromatic hydrocarbons. MO enzyme induction in the hepatic 13,000g fraction of male dab, measured as 7-ethoxyresorufin-O-deethylase activity, was significantly enhanced in response to low ambient temperatures. Some evidence was found for the facilitation of benzo[a]pyrene hydroxylase activity expressing the enzyme induction in the microsomal fraction of pyloric caeca of seastars, at increasing PCB concentrations. DNA integrity and enzyme induction elucidate the physiologic status and might be indicative for ambient impairment within restricted areas, and not necessarily related to the presence of anthropogenic or xenobiotic substances.

Animals↗

NADPH-, NADH- and cumene hydroperoxide-dependent metabolism of benzo[a]pyrene by pyloric caeca microsomes of the sea star Asterias rubens L. (Echinodermata: Asteroidea).

1. Benzo[a]pyrene (BaP) metabolism was studied in microsomes of the pyloric caeca (main digestive tissue and site of P450) of the echinoderm sea star (starfish) Asterias rubens. 2. NADPH-dependent metabolism of BaP produced phenols (36% of total metabolism), quinones (19%), dihydrodiols (25%) and putative protein adducts (20%). 3. NADH-dependent rates of BaP metabolism were approximately twice those found for NADPH-dependent metabolism, and metabolite formation was shifted towards dihydrodiols and quinones. 4. Cumene hydroperoxide (CHP)-dependent rates of BaP metabolism were also higher than NADPH-dependent rates by a factor of six for quinone and putative protein adduct production, and by a factor of four for phenol and dihydrodiol production. 5. Microsomal rates of BaP metabolism in BaP-exposed sea stars appeared to be elevated more in the case of NADPH-dependent than for CHP-dependent metabolism (respectively, increases of 130 and 41%), indicating the induction of forms of P450 preferentially catalysing NADPH-dependent metabolism. 6. 1,1,1-Trichloropropene-2,3-oxide (TCPO) inhibited dihydrodiol formation from both NADPH- and CHP-dependent BaP metabolism, indicating the involvement of epoxide hydratase in BaP metabolism. 7. Incubations of pyloric caeca microsomes with BaP and a superoxide anion radical-generating system (xanthine/xanthine oxidase) produced putative protein adducts but no free metabolites.

Animals↗

Effects of cadmium on metal composition and adenylate energy charge in the sea star Asterias rubens L.

Sea stars, Asterias rubens, were exposed to 200 micrograms Cd/L or fed with mussels which contained about 70 micrograms Cd/g dry wt. After 5 weeks, cadmium in the pyloric caeca of directly and indirectly exposed sea stars had reached levels of 12 and 9 micrograms Cd/g dry wt, respectively. For both types of exposure, a reduction of 30% of the zinc levels in the pyloric caeca was found, which was correlated with a comparable displacement of zinc from the metallothionein-like proteins. Copper levels were increased in the pyloric caeca of directly exposed sea stars. In gonads, stomachs, and body wall of directly exposed sea stars, cadmium concentrations were 4 to 9 times higher than those in animals fed with Cd-contaminated mussels. Cadmium exposure also affected metal composition in these tissues. The ovaries contained relatively large amounts of zinc. Gel filtration chromatography revealed that this zinc and the accumulated cadmium were distributed over a large range of high-molecular-weight proteins. Both direct and indirect cadmium exposure resulted in a small, but significant decrease of the adenylate energy charge (AEC) in the pyloric caeca. In the gonads, no effect of the cadmium exposure could be demonstrated on the AEC, but in the ovaries a reduction of the adenylate pool was found. In semi-field experiments, stars were exposed to 25 micrograms Cd/L or fed with mussels collected from the heavily polluted Dutch Western Scheldt. After 6 months of direct or indirect exposure, cadmium in the pyloric caeca had reached comparable levels of 8 and 7 micrograms/g dry wt, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of cadmium and PCBs on reproduction of the sea star Asterias rubens: aberrations in the early development.

In semifield experiments sea stars, Asterias rubens, were exposed to 25 micrograms Cd/liter or fed with mussels containing 0.6 microgram/g wet wt PCBs (Clophen A50). After 5 months of exposure, Cd concentrations in testes and ovaries were respectively 17 and 50 times higher than those in unexposed sea stars. PCB concentrations were respectively 7 and 9 times higher. With spermatozoa obtained from Cd- or PCB-exposed sea stars, normal fertilization could be achieved. However, maturation of oocytes from Cd-exposed animals was delayed and early development of embryos from Cd- or PCB-exposed animals was disturbed. Due to aberrations during the early development only 24 and 30% of the embryos obtained from Cd- or PCB-exposed sea stars, respectively, had developed to normal bipinnaria larvae after 1 week.

Animals↗

Effects of cadmium and zinc on steroid metabolism and steroid level in the sea star Asterias rubens L.

Steroid metabolism was studied in gonads and pyloric caeca of male and female sea stars which had been exposed to cadmium or zinc for 3 weeks. Steroid metabolism had increased in animals exposed to heavy metals. Significant increase of the enzyme activity was observed for 17 alpha-hydroxylase in the pyloric caeca of female animals exposed to zinc (pregnenolone----17 alpha-hydroxypregnenolone) or cadmium (progesterone----17 alpha-hydroxyprogesterone), for 17 beta-hydroxysteroiddehydrogenase in the gonads of female animals (dehydroepiandrosterone----androstenediol) and in pyloric caeca of male animals (androstenedione----testosterone) after exposure to cadmium, and for 5 alpha-reductase (progesterone----5 alpha-pregnane-3,20-dione) in ovaries of cadmium exposed sea star. There is some evidence that pregnenolone metabolism in male and female animals is affected by zinc in a different way. The effect of cadmium on the esterification of androstenedione differed highly significantly among male and female animals. It was concluded that the main way of entrance of cadmium into sea stars is via the surrounding medium and not via the food consumed. Testosterone and progesterone levels in, respectively, gonads of female and pyloric caeca of male sea stars which had been exposed to cadmium were significantly higher than the corresponding values in control animals. The effect of zinc exposure on testosterone level in pyloric caeca is significantly different for both sexes. Cadmium interacts directly with the esterification of testosterone, strongly stimulating this process. Cadmium stimulates the production of testosterone by action at the level of the biosynthesis of 17 beta-hydroxysteroiddehydrogenase.

17-Hydroxysteroid Dehydrogenases↗

In vivo exposure of Dreissena polymorpha mussels to the quinones menadione and lawsone: menadione is more toxic to mussels than lawsone.

The principal aim of this study was to assess whether the two quinones, menadione (2-methyl-1,4-naphthoquinone) and lawsone (2-hydroxy-1,4-naphthoquinone), elicit differential toxicity in mussels as has been reported for higher organisms. Therefore, the effects of short-term (48 h) and long-term (20 days) exposure of the two quinones at concentrations of 0.56 and 1 mg l(-1) to zebra mussels, Dreissena polymorpha, under laboratory conditions were studied. After the short-term exposure, the specific activities of the two-electron quinone oxidoreductase (DT-diaphorase) and the one-electron catalysing quinone reductases NADPH-cytochrome c reductase and NADH-cytochrome c reductase were determined in the gills and the rest of the soft tissues (soft mussel tissues minus the gills) of both treated and control mussels. At the higher concentrations of menadione and lawsone used, a significant reduction of the activity of NADPH-cytochrome c reductase in the gills and in the rest of the soft mussel tissues (by 33-34% and 31-43%, respectively) was observed. The activities of DT-diaphorase and NADH-cytochrome c reductase were not significantly affected. Interestingly, DT-diaphorase was observed in the gills, an organ requiring protection against antioxidants. Furthermore, a single-cell electrophoretic assay (comet assay) performed with gill cells to assess DNA damage by the quinones did not show any significant difference between the treated and the control organisms. This indicates that the formation of reactive species by the quinone metabolism in vivo in the mussels was possibly suppressed through the concerted action of DT-diaphorase and antioxidant enzymes. The results of in vitro experiments with gill extracts confirmed the protective role of DT-diaphorase. The rate of the two-electron quinone reduction was found to be five times that of the one-electron quinone reduction. The results of the long-term exposure unambiguously demonstrated that in mussels menadione, unlike in higher organisms, is more toxic than lawsone. The lack of detectability of xanthine oxidase in the mussel tissues could explain the comparatively lower toxicity of lawsone in the invertebtrate, lending support to a previous suggestion that xanthine oxidase might be responsible for the mechanism of toxicity of lawsone in higher organisms in vivo.

Animals↗

Relation between responses in the neutral red retention test and the comet assay and life history parameters of Daphnia magna.

Responses of the neutral red retention (NRR) assay as test for lysosomal stability and the comet assay as test for DNA integrity were measured in the water flea, Daphnia magna, and compared with mortality and effects on population growth rate during short- or long-term exposure to seven different toxicants. The NRR test and the comet assay were performed with fresh preparations of pieces of tissue from the digestive tract or with cell preparations from whole daphnias. Five toxicants caused responses of the NRR test or the comet assay after short-term exposure at concentrations below the acute toxicity level. Preliminary results of long-term exposure experiments suggest that these biomarker responses can be related to chronic effects on survival and/or reproduction of D. magna. This type of research should provide the basis for future use of the NRR test and the comet assay as early warning biomarkers for effects of toxicants on Daphnia populations.

Animals↗