The zebrafish embryo model in environmental risk assessment—applications beyond acute toxicity testing

BACKGROUND, AIM, AND SCOPE: The use of fish embryos is not regulated by current legislations on animal welfare and is therefore considered as a refinement, if not replacement of animal experiments. Fish embryos represent an attractive model for environmental risk assessment of chemicals since they offer the possibility to perform small-scale, high-throughput analyses. MAIN FEATURES: Beyond their application for determining the acute toxicity, fish embryos are also excellent models for studies aimed at the understanding of toxic mechanisms and the indication of possible adverse and long-term effects. Therefore, we have reviewed the scientific literature in order to indicate alternative applications of the fish embryo model with focus on embryos of the zebrafish. RESULTS AND DISCUSSIONS: The analysis of the mode of action is important for the risk assessment of environmental chemicals and can assist in indicating adverse and long-term effects. Toxicogenomics present a promising approach to unravel the potential mechanisms. Therefore, we present examples of the use of zebrafish embryos to study the effect of chemicals on gene and protein patterns, and the potential implications of differential expression for toxicity. The possible application of other methods, such as kinase arrays or metabolomic profiling, is also highlighted. Furthermore, we show examples of toxicokinetic studies (bioconcentration, ABC transporters) and discuss limitations that might be caused by the potential barrier function of the chorion. Finally, we demonstrate that biomarkers of endocrine disruption, immune modulation, genotoxicity or chronic toxicity could be used as indicators or predictors of sub-acute and long-term effects. CONCLUSIONS: The zebrafish embryo represents a model with an impressive range of possible applications in environmental sciences. Particularly, the adaptation of molecular, system-wide approaches from biomedical research is likely to extend its use in ecotoxicology. RECOMMENDATIONS AND PERSPECTIVES: Challenges for future research are (1) the identification of further suitable molecular markers as indicators of the mode of action, (2) the establishment of strong links between (molecular) effects in short-term assays in embryos and long-term (toxic) effects on individuals, (3) the definition of limitations of the model and (4) the development of tests that can be used for regulatory purposes.     

Fate of cancerostatic platinum compounds in biological wastewater treatment of hospital effluents

The present work focuses on the fate of two cancerostatic platinum compounds (CPC), cisplatin and carboplatin, as well as of two inorganic platinum compounds, [PtCl₄]²⁻ and [PtCl₆]²⁻ in biological wastewater treatment. Laboratory experiments modelling adsorption of these compounds onto activated sludge showed promising specific adsorption coefficients K_D and K_OC and Freundlich adsorption isotherms. However, the adsorption properties of the investigated substances were differing significantly. Adsorption decreased following the order cisplatin > [PtCl₆]²⁻ > [PtCl₄]²⁻ > carboplatin. Log K_D-values were ranging from 2.5 to 4.3 , log K_OC from 3.0 to 4.7.

A pilot membrane bioreactor system (MBR) was installed in a hospital in Vienna and fed with wastewater from the oncologic in-patient treatment ward to investigate CPC-adsorption in a sewage treatment plant. During three monitoring periods Pt-concentrations were measured in the influent (3–250 μg l⁻¹ Pt) and the effluent (2–150 μg l⁻¹ Pt) of the treatment plant using ICP-MS. The monitoring periods (duration 30 d) revealed elimination efficiencies between 51% and 63% based on averaged weekly input–output budgets. The derived log K_D-values and log K_OC-values ranged from 2.4 to 4.8 and from 2.8 to 5.3, respectively. Species analysis using HPLC-ICP-MS proofed that mainly carboplatin was present as intact drug in the influent and – due to low log K_D – in the effluent of the MBR.     

The connectedness to nature scale: A measure of individuals’ feeling in community with nature

Five studies assessed the validity and reliability of the connectedness to nature scale (CNS), a new measure of individuals’ trait levels of feeling emotionally connected to the natural world. Data from two community and three college samples demonstrated that the CNS has good psychometric properties, correlates with related variables (the new environmental paradigm scale, identity as an environmentalist), and is uncorrelated with potential confounds (verbal ability, social desirability). This paper supports ecopsychologists’ contention that connection to nature is an important predictor of ecological behavior and subjective well-being. It also extends social psychological research on self–other overlap, perspective taking, and altruistic behavior to the overlap between self and nature. The CNS promises to be a useful empirical tool for research on the relationship between humans and the natural world.     

Wirkung subakuter PCB-Exposition (Aroclor 1254) auf Sauerstoffverbrauch, Schwimmbewegung und Biotransformation (GST-Aktivität) des Karpfens (Cyprinus carpio)

Objective and Background

PCB (polychlorinated biphenyle) are typical man-made environmental pollutants that cause a broad spectrum of effects in vertebrates. Although a lot has been studied about the toxic effects of xenobiotic substances on aquatic organisms, their subacute effects are not yet well known. Fish occupies a central position in freshwater food webs and therefore, carp was chosen to be our test organism. The aim of our study was the investigation of subacute effects of Aroclor 1254 (22μg ^l?1) on respiration, swimming activity, and biotransformation, allowing discussion of changed interaction between the available energy resources.

Methods

The respiration experiments under controlled laboratory conditions were run for 29 days (5 d before, 16 d with and 8 d after chemical exposure) with exposed carp compared to the non-exposed fish. Furthermore, the BehavioQuant system quantitatively monitors the positions of each individual fish before (8 d) and during (21 d) PCB exposure and swimming activity (number of horizontal turnings s^?1) of animals was calculated. Thereafter, liver samples were taken from animals for analysis of the phase 2 enzyme activity (glutathione-S-transferase). In the respiration experiments were 6 exposure, 6 controll, and 3 solubilizer controll groups (12 animals each group); in the behavioral experiments were 9 exposure, 6 controll- und 3 solubilizer controll groups (6 animals each group), and in the enzyme activity measurements were 9 exposure, 6 controll and 3 solubilizer controll groups (6 animals each group) examined.

Results and Discussion

A clearly increased (33%) oxygen consumption due to the PCB-exposure is found to be reversible; after the exposure period the respiration of carp shows a level comparable to the oxygen consumption during non-exposure time. By evaluating quantitative behavioral parameters of animals, it becomes evident that the exposure to PCB also causes a reversible change in their swimming behavior. Chemical stress leads to a decrease of mean daily swimming activity combined with an increase of the mean number of turnings during the night. Thereafter, the swimming activity shifts to the circadian swimming behavior under control conditions. Furthermore, our measurement of soluble and microsomal glutathione-S-transferase activity of fish liver shows a significant elevation after exposure period.

Conclusion

Our results prove that it was feasible to detect sublethal effects of PCB-mixture on all parameters under these conditions successfully. Our findings imply that fish are able to cope with the chemicals and we purse the hypothesis that the higher physiological energy demand caused by increased activity of enzymes, which are involved in biotransformation of foregin substances, may be related to a higher respiration of fish stressed by xenobiotics.

Recommendation and Outlook

Further experiments should investigate the effects of PCB on ecophysiological parameters of carp fed with exposed natural food allowing discussion of ecological consequences.     

Aphid alarm pheromone (E)-β-farnesene: A host finding kairomone for the aphid primary parasitoidAphidius uzbekistanicus (Hymenoptera: Aphidiinae)

Summary The present study aimed to test the possible function of the aphid alarm pheromone (E)--farnesene (EBF) as a host finding kairomone for aphid primary parasitoids. Extracts of volatile emissions of undisturbed aphids and of aphids under parasitoid attack were obtained by air entrainment. Extracts of cornicle secretions were gained by disturbing aphids and taking their secretions into solution. Extracts were compared by gas chromatography. Only air entrainments of aphids under attack and solvent extracts of cornicle secretions contained the alarm pheromone. In Y-tube olfactometer bioassays, femaleA. uzbekistanicus were attracted to aphid groups under attack of parasitoids, presumably by released EBF. High concentrations of synthetic EBF (1.4 µg to 5.7 µg) also attractedA. uzbekistanicus females. Females with oviposition experience reacted more readily to lower concentrations of EBF than females without experience. In experiments designed as Petri-dish bioassays, the test animals could contact filter paper discs that were treated with solutions containing EBF. Behavioural effects like antennation or stinging attack were not observed. With computer video analysis of parasitoid movements, some effects onA. uzbekistanicus behaviour were detected, again indicating attraction to EBF.As the volatile aphid alarm pheromone attractedA. uzbekistanicus females, it can be concluded that this chemical stimulus acts as a host finding kairomone for this parasitoid species. However, its effect over long distances seems to be limited due to the relatively high concentrations required for reactions. Of two other parasitoid species examined (P. volucre andL. testaceipes) onlyP. volucre was also significantly attracted to the volatile EBF in the Y-tube olfactometer.     

Terrestrische ökotoxikologische Testmethoden für die Tropen

Goal, Scope, and Background

Soil organisms play a crucial role in the terrestrial ecosystem. Plant Protection Products (PPPs) are known to affect soil organisms and might have negative impacts on soil functions influenced by these organisms. Little research has been done to day on the impact of PPs on tropical ecosystems. Therefore, in this study it was investigated whether fate and effects of pesticides differ between tropical and temperate regions and whether data generated under temperate conditions can be used for the Environmental Risk Assessment (ERA) in tropical regions.

Methods

In the first part of this study, the effects of two fungicides (Benomyl and Carbendazim) and one insecticide (lambda-Cyhalothrin) on soil invertebrates (i.e. earthworms and arthropods) were evaluated in laboratory tests modified for tropical conditions (temperature, soil, test species). Besides using some native species, the tests were done mainly with two (temperate and tropical) strains of earthworms (Eisenia fetida) and the peregrine isopod speciesPorcellionides pruinosus as standard test species. The chemicals were spiked in two natural and two artificial soils. A tropical artificial soil (TAS), containing a tropical fern product (Xaxim) or coconut coir dust as organic matter, was developed in this study.

Results and Conclusions

The results from the laboratory tests showed that all three test chemicals differed from those gained under temperate conditions. In the case of the fungicides the toxicity was lower but in the case of the insecticide higher under tropical than under temperate conditions. The native tropical earthwormPontoscolex corethrurus reacted more sensitively against Carbendazim in comparison to the standard test speciesEisenia fetida.

Recommendation and Perspective

Details of the environmental risk assessment of the three model chemicals based on the results of the laboratory described here (and including the results of higher tier tests (semi-field and field tests)) will be described in Part 2 of this series     

Fate of large-scale vortices in idealized tidal lagoons

Environmental Fluid Mechanics - The generation and evolution of tidally-induced vortices in coastal and estuarine regions can influence water quality and sedimentary processes. These effects must...     

A simulator study of driving behavior and mental workload in mixed-use arterial road environments

Objectives: Mixed-use urban environments, such as arterial roads with adjacent commercial land uses, represent crash locations with the highest risk. These locations are often characterized by high volumes of motor vehicle traffic, on-street parking, and interactions with multiple road user groups such as pedestrians, cyclists, and public transportation. The objective of this study was to investigate previously identified crash risk factors for mixed-use urban environments and assess how parking occupancy, center medians, and cyclist volume influence performance and workload in a driving simulator study.

Methods: Thirty participants were recruited for the study. Participants completed 6 drives that presented different combinations of cyclist volume, median condition, and parking occupancy. Incorporated into the simulator drives was a secondary peripheral detection task (PDT) designed to measure mental workload. Participants provided subjective assessments of workload using the Rating Scale Mental Effort (RSME).

Results: Mean lateral lane position was found to significantly vary across the 3 independent variables of parking occupancy, cyclist volume, and median conditions. No significant changes were identified for mean speed across the conditions. Subjective and objective measures of workload identified changes due to the presence of cyclists with slower reaction times for the PDT task when cyclists were present.

Conclusion: The findings provide insight into the interaction of road design elements in mixed-use urban road environments and demonstrate that increasingly complex environments increase driver demand. This has important road design implications for mixed-use arterial roads, which are often characterized by complex interactions between multiple road user groups.     

Conservation impacts of commercial cultivation of endangered and overharvested plants

Overharvesting is one of the greatest threats to species survival. Farming overharvested species is a conservation strategy that can meet growing market demand and conserve wild populations of the target species. This strategy is compatible with the international community's desire to uphold the right of local communities to use biological resources to support their livelihoods. However, studies investigating whether farming can alleviate poaching pressure have focused almost exclusively on animals. To address the shortfall in plant-focused studies, we compiled information on commercial cultivation of threatened plants to assess its conservation benefits. Because China's rising middle class has rapidly intensified demand for wildlife products, we searched the scientific literature published in Chinese (China National Knowledge Infrastructure and Baidu) and in English. We found 32 reports that contained data on 193 internationally or nationally threatened plant species that were under commercial cultivation. These reports showed that cultivations of 82% of the 193 species were sustained by collecting whole plants from the wild periodically or continuously. Although based on a small sample size, species that were maintained in cultivation only through artificial propagation or seeds collected in the wild were likely associated with a reported reduction in wild harvesting of whole plants. Even so, results of correlation analyses suggested that production system, scale, and when a species began being cultivated had little effect on conservation status of the species, either globally or in China. However, species brought into cultivation relatively recently and on a smaller scale were more likely to have undergone a reduction in collecting pressure. Farming of nonmedicinal plants was most problematic for species conservation because wild plants were laundered (i.e., sold as cultivated plants). For effective conservation, policy to guide cultivation operations based on the target species’ biological characteristics, cultural significance, market demand, and conservation status is needed.