Ulrik Nørum

Vitellogenin induction by 17β-estradiol and 17α-ethinylestradiol in male zebrafish (Danio rerio)

Adult male zebrafish (Danio rerio) were exposed to 17beta-estradiol (E2) or 17alpha-ethinylestradiol (EE2) in flow-through systems for 8 days. This was done to compare the sensitivity of the estrogen inducible vitellogenin (Vtg) biomarker system of this proposed OECD test guideline species to other relevant test species. Vtg was quantified in whole body homogenate by a species-specific ELISA. Actual water concentrations of E2 and EE2 were quantified by LC-MS, with detection limits of 1.0 and 0.6 ng/l, respectively. Vtg induction (LOEC) occurred in whole body homogenate at actual water concentrations of 21 ng E2/l and 3.0 ng EE2/l, respectively. As an alternative to the ANOVA approach, the relationship between the percentage of responding fish (Vtg) and the external E2 or EE2 concentration was determined by logistic regression analysis. Based on the regression analysis, EC-values could be determined: EC10, EC50 and EC90 were 15.4, 41.2 and 67.1 ng E2/l, respectively and 0.92, 2.51 and 4.09 ng EE2/l, respectively. Comparisons of these response limits to corresponding values for rainbow trout (Oncorhynchus mykiss), fathead minnow (Pimephales promelas) and Japanese medaka (Oryzias latipes) revealed the zebrafish as a sensitive test species.

Behavioural changes in three species of freshwater macroinvertebrates exposed to the pyrethroid lambda-cyhalothrin: laboratory and stream microcosm studies.

Pesticides are transported from crop fields to adjacent streams via surface run-off, drains, groundwater, wind drift and atmospheric deposition and give rise to transient pulse contamination. Although the concentrations observed, typically <10 microg L(-1), cannot be expected to be acutely lethal, effects in streams at the population and ecosystem level have been reported. One of the most conspicuous phenomena associated with these transient pesticide pulses is drift, where large numbers of freshwater invertebrates are carried along by the current and disappear from the contaminated stretch of the stream. The aim of the present study was to evaluate the feasibility of linking laboratory studies of the sublethal effects of pulse exposure to the pyrethroid lambda-cyhalothrin on the locomotory behaviour of stream invertebrates with effects on drift behaviour under more environmentally realistic conditions in stream microcosms. In the laboratory as well as in the microcosms, the order of sensitivities of the three species tested was (with Leuctra nigra being the most sensitive): L. nigra>Gammarus pulex>Heptagenia sulphurea. The LOECs determined for L. nigra (1 ng L(-1)), G. pulex (10 ng L(-1)) and H. sulphurea (100 ng L(-1)) are all within expected environmental concentrations. For the species of invertebrates investigated, it was possible to extrapolate directly from pyrethroid-induced behavioural changes observed in the laboratory to drift under more realistic conditions in stream microcosms. Consequently, the fast and cost-effective video tracking methodology may be applied for screening for potential effects of a wider range of pesticides and other stressors on the locomotory behaviour of freshwater invertebrates. The results indicate that such behavioural changes may be predictive of effects at the ecosystem level.

Future migratory behaviour predicted from premigratory levels of gill Na+/K+-ATPase activity in individual wild brown trout (Salmo trutta)

SUMMARY The relationship between premigratory gill Na+/K+-ATPase activity, determined at two dates during spring, and future migratory behaviour was investigated using non-lethal gill biopsies and PIT-tagging in wild brown trout (Salmo trutta) from two tributaries. No significant relationship between future migratory strategy (individuals eventually becoming migrants or residents) and gill Na+/K+-ATPase activity was found in late February–early March. By contrast, in mid-April, a highly significant logistic regression equation identifying the migratory strategy in 93% of the 75 individuals was obtained. The ability of this regression model from the tributaries to predict future migratory behaviour in an independent group of trout caught in early April in the mainstream was evaluated. A threshold probability of migration was used to predict the behaviour of the mainstream individuals as either future migrants or residents. The maximum percentage of correct predictions of future migratory behaviour in mainstream fish was observed at threshold probabilities between approximately 0.15 and 0.45 (corresponding to threshold gill Na+/K+-ATPase activities between 2.7 and 3.7 μmol ADP mg–1 protein h–1), with an average of 91% of the predictions being correct. The present study shows that a non-lethal premigratory biochemical measurement can successfully select individual brown trout with high probability of migration.

Pesticide impacts on predator–prey interactions across two levels of organisation

In this study, we aimed to evaluate the effects of a short pulse exposure of the pyrethroid lambda-cyhalothrin (LC) on the predator and anti-predator behaviour of the same species; Gammarus pulex. Predator behaviour, at the level of the individual, was studied in indoor microcosms using video tracking equipment during simultaneous exposure of the predator (G. pulex) and its prey (Leuctra nigra) during 90 min exposure of 1, 6.6 or 62.1 ngL(-1) LC. During an initial 30 min of exposure, the predator and prey organisms were maintained physically separated, and the actual interaction was studied through the subsequent 60 min of exposure. The anti-predator behaviour of G. pulex (drift suppression in response to the presence of brown trout) was studied in outdoor stream channels during a 90 min pulse exposure to LC (7.4 or 79.5 ngL(-1)) with, or without, brown trout. Based on survival curves for L. nigra we found that the mortality rate for L. nigra significantly decreased during exposure to 6.6 and 62.1 ngL(-1) LC (P<0.05 and P<0.001, respectively). We found no significant effects suggesting that G. pulex was repelled by contaminated prey items (P>0.05). We found that the exposure of G. pulex to 7.4 and 79.5 ngL(-1) LC significantly increased drift (from ∼0% to ∼100% in both treatments; P<0.001) independent of the presence of brown trout (P<0.05). In other words, the natural anti-predator behaviour of G. pulex was overruled by the stress response to LC exposure increasing G. pulex predation risk from drift feeding brown trouts. Our results show that the anti-predator and predator behaviour of G. pulex were significantly changed during exposure to very low and environmentally realistic LC concentrations and exposure duration. The potential implications for the field scenario are discussed.

New model for gain control of signal intensity to object distance in echolocating bats

SUMMARY Echolocating bats emit ultrasonic calls and listen for the returning echoes to orient and localize prey in darkness. The emitted source level, SL (estimated signal intensity 10 cm from the mouth), is adjusted dynamically from call to call in response to sensory feedback as bats approach objects. A logarithmic relationship of SL=20log10(x), i.e. 6 dB output reduction per halving of distance, x, has been proposed as a model for the relationship between emitted intensity and object distance, not only for bats but also for echolocating toothed whales. This logarithmic model suggests that the approaching echolocator maintains a constant intensity impinging upon the object, but it also implies ever-increasing source levels with distance, a physical and biological impossibility. We developed a new model for intensity compensation with an exponential rise to the maximum source level: SL=SLmax–ae–bx. In addition to providing a method for estimating maximum output, the new model also offers a tool for estimating a minimum detection distance where intensity compensation starts. We tested the new exponential model against the ‘conventional’ logarithmic model on data from five bat species. The new model performed better in 77% of the trials and as good as the conventional model in the rest (23%). We found much steeper rates of compensation when fitting the model to individual rather than pooled data, with slopes often steeper than –20 dB per halving of distance. This emphasizes the importance of analyzing individual events. The results are discussed in light of habitat constraints and the interaction between bats and their eared prey.

Arsenic compounds in the haemolymph of the Dungeness crab, Cancer magister, as determined by using HPLC on-line with inductively coupled plasma mass spectrometry

Arsenobetaine, two arsenosugars, dimethylarsinate and several unidentified arsenic species were detected in extracts of the haemolymph of the Dungeness crab, Cancer magister, by using HPLC-ICP-MS. This is the first report of the presence of arsenosugars in the haemolymph/blood of marine animals. Total, extractable and residual arsenic concentrations were determined by ICP-MS. The concentration of total arsenic was in the range of 1.4-3.8 [micro sign]g ml(-1). Nearly all (98%) the arsenic was found to be extractable, and accounted for primarily by arsenobetaine, two arsenosugars and dimethylarsinate. The results demonstrate that arsenic compounds present in the diet of crabs are not fully metabolized in the gut. They are, at least partly, taken up into the haemolymph. The concurrence of arsenobetaine and arsenosugars suggests that the use of repeated haemolymph sampling in crustaceans could facilitate investigations into the kinetics of the biotransformation pathways of arsenic compounds. Finally, the present study clearly demonstrates the unique capabilities of HPLC-ICP-MS for the detection and identification of minor arsenic components amongst the predominant arsenobetaine.