Automated beta galactosidase activity bioassay for adult Daphnia magna versus classic immobilization test.
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Biomedical subjects
Publications and source records attributed to R Weltens.
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It is generally assumed that the dissolved fraction of a toxic substance in surface water is mainly responsible for toxicity to aquatic organisms. However, toxic compounds are often adsorbed or chemically bound to suspended particles in the water column, depending upon the physico-chemical conditions. In the present study potential adverse effects to filter feeding organisms by metal contaminated particles were investigated. In our hypotheses the adsorbed metals might desorb in the gastrointestinal tract-due to different physico-chemical conditions-and exert toxic effects. Clay and sand particles, algae and organic material (peat) were artificially contaminated with cadmium and zinc. The contaminated materials were resuspended in standard conditions and toxicity was measured for the water flea Daphnia magna (mortality at 48 hours). As a reference, supernatant solutions were used containing the same concentration of dissolved metal as the suspensions. It was also established that the test concentrations of solid material (250 and 500 mg/l uncontaminated particles) did not cause any mortality within 48 hours. Daphnids are filter feeders: they filtrate large amounts of surrounding water, redrawing particles as a food source. Results strongly indicate that contaminated particles threaten the health of these particle-feeding organisms. Compared to the reference severe acute toxic effects were seen and cadmium accumulation was increased when contaminated solid material was present. Results were essentially the same for the different materials used in the experiments, except for sand contaminated with cadmium. This shows that mineral as well as organic materials can contribute to the particle bound toxicity. Different results were obtained when a static set up was used instead of a flow through set up, illustrating that the route of administration is important to make particles available and thus to evaluate their toxicity. Contaminated particles clearly have toxic potency, not only because they are a continuous source of dissolved xenobiotics, but also because the particle bound fraction can become available within the body of particle feeding organisms. This could lead to unexpected high tissue concentrations. More insight is needed to predict the bioavailability of adsorbed pollutants. Results of this study already indicate that suspended solids should be considered as a separate compartment in risk evaluation of chemicals, effluents or natural surface waters.
Increasing evidence suggests that the use of a single bioassay will never provide a full picture of the quality of the environment. Only a test battery, composed of bioassays of different animal and plant species from different trophic levels will reduce uncertainty, allowing an accurate assessment of the quality of the environment. In the present study, a test battery composed of 20 bioassays of varying biological endpoints has been compared. Apart from lethality and reproductive failure in earthworms, springtails, nematoda, algae and vascular plants, these endpoints also included bioavailibility of metals (bacteria), heat-shock induction (nematodes, algae), DNA damage (bacteria, earthworm, vascular plants), beta-galactosidase (Daphnia) and esterase activity (algae) and a range of immunological parameters (earthworm). Four chemicals (cadmium, phenol, pentachlorophenol and trifluralin)--each representing a different toxic mode of action--were applied in a dilution series (from 1 mg/kg up to 1000 mg/kg) onto OECD standard soil. The tests have been performed both on these artificially contaminated soil samples and on aqueous extracts subsequently obtained from these soils. The results show that the immunological parameters and the loss of weight in the earthworms were among the most sensitive solid-phase assays. Esterase inhibition and heat-shock induction in algae were shown to be extremely sensitive when applied to soil extracts. As previously shown at the species level, no single biological endpoint was shown to be the most sensitive for all four modes of toxic action.
Cellular and luminal pH of isolated ant Malpighian tubules were measured in different bath K+ concentrations using double-barrelled pH microelectrodes. The electrochemical gradient for H+ across the basolateral and the apical cell membranes was estimated. In control Ringer (51 mmol/l K+) cell and luminal pH were alkaline with respect to the basolateral solution: 7.77 and 7.36, respectively, versus 7.25. On lowering basolateral K+ concentration to 5 mmol/l or increasing it to 113 mmol/l, luminal pH and to a lesser extent cell pH followed: luminal pH changed to 7.14 and 7.43 and cell pH to 7.69 and 7.82, respectively. In all conditions a cell inward electrochemical gradient for protons across both membranes was observed. Increasing basolateral K+ concentration, which was positively correlated with secretion rate, decreased the cell inwardly directed apical proton gradient; moreover, the apical membrane potential difference decreased as well, from -93 mV in 5 mmol/l K+ to -65 mV in 113 mmol/l K+. Therefore the turnover rate of the electrogenic active proton pump at the apical membrane is facilitated in a high basolateral K+ concentration. The calculated electromotive force of this pump is -159 mV. Comparing the proton with the K+ electrochemical gradient, taken from another study in the same experimental conditions, we find that the apical proton electrochemical gradient can drive K+ extrusion into the lumen for each value of secretion rate.
The number of AgNOR (NOR+) and the amount of AgNOR (NORM+) were analysed by means of two multilevel analyses of variance in a total of 12 twin pairs: 3 female and 4 male MZ and 5 male DZ pairs. In the first analysis, only zygosity was controlled; in the second, chromosome types D and G were controlled as well as the interaction between chromosome type and zygosity. For NOR+ and NORM+, when chromosome types D and G are not distinguished, the within-pair variance is greater, though not significantly, in DZ than in MZ pairs; but it is highly significantly greater when chromosome type (D or G type) is under control. This confirms an important genetic determination of NOR+ and NORM+ when in the ANOVA model the D and G types are controlled. However, nongenetic factors also influence the Ag-NOR patterns, but not enough to conceal the genetically defined rDNA pattern. Indeed, about 50% of the cells transcribe their rDNA in a way not closely dependent on the rDNA background and significant intrapair differences of NOR+ pattern exist in MZ twins.