E.A.J. Bleeker

Comparative toxic and genotoxic effects of chloroacetanilides, formamidines and their degradation products on Vibrio fischeri and Chironomus riparius

Toxic and genotoxic effects of alachlor, metolachlor, amitraz, chlordimeform, their respective environmentally stable degradation products 2,6-diethylaniline, 2-ethyl-4-methylaniline, 2,4-dimethylaniline, and two other related compounds, 3,4-dichloroaniline and aniline were compared. Acute toxicity tests with Chironomus riparius (96 h) and Vibrio fischeri (Microtox) and genotoxicity tests with a dark mutant of V. fischeri (Mutato) were carried out. Our results demonstrate that toxicity and genotoxicity of the pesticides are retained upon degradation to their alkyl-aniline metabolites. In the case of the herbicides alachlor and metolachlor, the toxicity to V. fischeri was enhanced upon degradation. Narcosis alone explains toxicity of the compounds to the midge, but not so for the bacteria suggesting a disparity in the selectivity of the test systems. All compounds showed direct genotoxicity in the Vibrio test. but amitraz and its metabolite were genotoxic at concentrations 10(3)-10(5) lower than all the other compounds. The observations indicate that stable aniline degradation products of the pesticides may contribute considerably to environmental risks of pesticides application and that genotoxic effects may arise upon degradation of pesticides.

Comparative ecotoxicity of NPAHs to larvae of the midge Chironomus riparius

Abstract Acute toxicity of seven azaarenes (NPAHs) has been determined using larvae of the midge Chironomus riparius. Clear dose-response relationships for mortality of first instar larvae were observed, but surviving larvae grew as equally well as the controls. LC50 values (96 h) for a two-ringed structure (quinoline), four three-ringed structures (acridine, phenanthridine, benzo[f]quinoline and benzo[h]quinoline) and two four-ringed structures (benz[a]acridine and benz[c]acridine) were generally one order lower than the few values reported for other aquatic species in the literature. Dibenz[a,i]acridine, a five-ringed structure, was not acutely toxic at its maximum dissolved concentration. Acute toxicity increased with increasing number of aromatic rings of the compound; accordingly, correlations between the LC50 values and size, shape and topology-related molecular properties were strong. However, between the four benzoquinoline isomers tested also differences in toxicity were observed: acridine was significantly more toxic than the other benzoquinoline isomers tested. This is most likely caused by transformation of acridine under UV light, indicated by a HOMO–LUMO gap in the highly phototoxic region, a high correlation between isomer toxicity and heat of formation and a low recovery of acridine in the water during the experiment.

Chronic toxicity of polycyclic aromatic compounds to the springtail Folsomia candida and the enchytraeid Enchytraeus crypticus

An urgent need exists for incorporating heterocyclic compounds and (bio)transformation products in ecotoxicological test schemes and risk assessment of polycyclic aromatic compounds (PACs). The aim of the present study therefore was to determine the chronic effects of (heterocyclic) PACs on two terrestrial invertebrates, the springtail Folsomia candida and the enchytraeid Enchytraeus crypticus. The effects of 11 PACs were determined in chronic experiments using reproduction and survival as endpoints. The results demonstrated that as far as narcosis-induced mortality is concerned, effects of both homocyclic and heterocyclic PACs are well described by the relationship between estimated pore-water 50% lethal concentrations and log Kow. In contrast, specific effects on reproduction varied between species and between compounds as closely related as isomers, showing up as deviations from the relationship between pore-water 50% effect concentrations and log Kow. These unpredictable specific effects on reproduction force one to test the toxicity of these PACs to populations of soil invertebrates to obtain reliable effect concentrations for use in risk assessment of PACs.

Linking ecological and ecotoxicological techniques to support river rehabilitation.

Human activities in river catchments interfere with natural fluxes of water and materials. Diffuse inputs and point-sources of toxicants have modified the ecological state of riverine communities considerably, and sanitation schemes are now under development for various rivers. To improve analysis, monitoring and prospecting the role of toxicants in river ecosystems a review of the available methods is undertaken. Ecotoxicological techniques are discussed in relation to basic ecological principles that are thought to regulate the functioning of communities. The response to toxicants among species is highly diverse and therefore the choice of test species (e.g. of typical riverine insects as caddisflies or mayflies) is critical, as it is the use of test-batteries. Long-term exposure may lead to developmental disturbances that may be assessed through morphometric techniques like analysis of asymmetry. Multi-generation exposure, although rarely studied, provides a useful insight into the genetic consequences of pollution. Selection for tolerant species or varieties has been experimentally assessed for smaller organisms such as insects, micro-algae, and bacteria. There is also perspective for multivariate analysis of species distribution in relation to pollutant exposure. Furthermore, a system approach to benthic ecology and sediment testing is needed. Such an approach reflects the strong linkage of ecological and ecotoxicological processes. Toxicants are transformed by biological activity; in some cases this alleviates toxicant stress, but in other cases degradation products are toxic as well. The risk of transformation to mutagenic products in the environment is indicated. The re-assessment of some of the classical ecotoxicological techniques is needed to adequately fulfil the needs of ecological recovery programs. To this purpose integration of ecotoxicological and ecological tools is needed.

Formation and identification of azaarene transformation products from aquatic invertebrate and algal metabolism.

The metabolism of two azaarenes, viz. acridine and phenanthridine, by aquatic organisms was studied in short-term and chronic laboratory tests. The identity of metabolites observed in the test waters was investigated with different analytical methods, including HPLC, GC and hyphenated LC- or GC-MS. The Zebra mussel (Dreissena polymorpha), one green alga species (Selenastrum capricornutum) and periphyton or bacteria transformed acridine into 9[10H]-acridinone. Phenanthridine was transformed into 5[6H]-phenanthridinone by midge (Chironomus riparius) larvae. The findings indicate that closely related isomers may undergo species-specific biotransformation. It was concluded that keto-metabolites are major products in the aquatic fate of benzoquinolines, which may be overlooked in the risk assessment of parent compounds. This study illustrates the typical problems with, as well as the potency of, chromatographic methods in the elucidation of metabolic routes of organic contaminants.

Comparative metabolism of phenanthridine by carp (Cyprinus carpio) and midge larvae (Chironomus riparius).

Abiotic transformation of azaarenes in the environment has been analysed extensively, but metabolism is less well described. To further elucidate preliminary observations of interspecific differences in azaarene metabolism by aquatic organisms, phenanthridine biotransformation by midge larvae and carp was studied. In both experiments, 6(5H)-phenanthridinone (phenanthridone) was found as an important metabolite. The fish were clearly capable of metabolising phenanthridine, but in the midge experiment the metabolite was principally formed by bacteria growing on the food and not by the midges. Phenanthridone itself was further degraded to non-observed compounds in both experiments, due to bacteria and midges acting together in the midge experiment, and by carp in the fish experiment. Internal concentrations of phenanthridine and phenanthridone were non-detectable in the midge larvae, but concentrations of both compounds in carp organs suggested a major role of bile and liver. Since phenanthridone did not account for all phenanthridine loss, it was suggested that, apart from phenanthridone degradation, other metabolic pathways may play a role. This study clearly demonstrates the importance of interspecies differences in metabolism, which should not be neglected in risk assessment.