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.     

Crowding and Experience-Use History: A Study of the Moderating Effect of Place Attachment Among Water-Based Recreationists

Effective recreation resource management relies on understanding visitor perceptions and behaviors. Given current and increasing pressures on water resources, understanding crowding evaluations seems important. Beyond crowding, however, variables that possibly relate to or influence crowding are of interest and in particular, place attachment and experience-use history (EUH). As EUH is related to place attachment and likely affects crowding, this study explored the moderating effect of place attachment dimensions on the relationships between EUH and visitor crowding evaluations. Water based recreationists at a U.S. Army Corps of Engineers site were contacted onsite and asked questions related to experience-use history, crowding evaluations, place attachment, and activity participation. Anglers and campers at the site identified similar crowding perceptions and place attachments. Only one of eight models tested revealed a moderating effect. Specifically, place identity moderated the relationship between the total times visited in the past twelve months and expected crowding among anglers. As such, the quest continues to understand the relationship among these important variables.     

ASSESSMENT OF ANTHROPOGENIC LOADS ON LANDSCAPES AS A TOOL TO DETERMINE THE POTENTIAL FOR SUSTAINABLE REGIONAL DEVELOPMENT: CASE STUDY FROM AZERBAIJAN

In order to determine the potential for sustainable regional development of the Caspian coastal zone, a study was made for the Khachmaz–Absheron zone of Azerbaijan. An evaluation was performed to assess the anthropogenic load on landscapes. Using the 8-point scale offered by Isachenko [2001, Ecological Geography of Russia, Saint Petersburg University Press, Saint Petersburg] for the indicators of agricultural, industrial, urban and integral anthropogenic loads, we did a preliminary ranking of the provinces in the Khachmaz–Absheron zone of Azerbaijan for the each indicator taken separately. Vital statistics were used as a supplementary indicator of environmental conditions in the region. By comparing the data for provinces with each other and the data on Azerbaijan average, we have classified the provinces into 4 groups according to specific combinations of the indicators. Each group of provinces has distinctive environmental conditions and features for sustainable development. The classification makes it possible to develop certain recommendations for the regional sustainable development. Measures to be implemented within the Azerbaijan State Program on Social and Economic Development of Regions are also discussed. In the Khachmaz province, production of ecologically pure products is highly recommended. Special attention should be given to the development of tourism and recreational institutions in the Khachmaz, Khizi and Devechi provinces. Recommendations for these provinces include further development of industry on the basis of modern safe technologies. The irrigation and drainage networks should be reconstructed. The reconstruction will make it possible to reduce water loss and to increase the productivity of agriculture. In the Absheron province, Baku and Sumgait cities environmental systems are overloaded, and so the works at highly polluting enterprises must be stopped, the enterprises have to be re-equipped (old filters must be changed first of all) or relocated from the area (a decrease of environmental risk should be in the focus of attention). For improving of air quality, green areas should increase. There is a need to reduce urban traffic density and to reconstruct highways. Use of old vehicles must be forbidden or restricted, and transition to environmentally friendly fuel should be supported in every way possible. It is necessary to bring the waste management system and sanitary landfills up to international standards, and to improve the water supply and sewerage systems.     

Dioxin formation in pulp and paper mills of India

Samples of effluents, sludge, pulp, final products (paper) and soil were collected from the identified pulp and paper mills in India. The samples were analysed for 2,3,7,8-tetrachloro-dibenzo-p-dioxin (2,3,7,8-TCDD) and other dioxin congeners and precursors. Pulp and paper mills using chlorine for the bleaching process showed the presence of 2,3,7,8-TCDD in effluent samples. In the effluent and pulp samples from mills where chlorine dioxide was used as a bleaching agent, the 2,3,7,8-TCDD congener ranged from below the detection limit 0.05 to 0.12 ngL⁻¹/ngg⁻¹. The relative standard deviation of reproducibility and the percent recovery of 2,3,7,8-TCDD were 2.07 and 82.4% in pulp and 2.8 and 92% in effluent, respectively. The 1,3,6,8-TCDD was the only other major dioxin congener found in the treated and untreated effluent and sludge samples. However, dichlorobenzene, trichlorophenyl, and hexachlorobiphenyl were detected in all samples. The formation of dioxins can be minimised by replacing chlorine with chlorine dioxide in bleaching processes in pulp and paper mills.     

Methane and carbon dioxide emission in a two-phase olive oil mill sludge windrow pile during composting

The aim of this work was to make some preliminary evaluations on CO(2) and CH(4) emissions during composting of two-phase olive oil mill sludge (OOMS). OOMS, olive tree leaves (OTL) and shredded olive tree branches (OTB) were used as feedstock for Pile I and Pile II with a 1:1:1 and 1:1:2v/v ratio, respectively. Each pile was originally 1.2m high, 2.0m wide and approximately 15.0m long. Four 500 ml volume glass funnels were inverted and introduced in each pile, two in the core (buried 50-60 cm from the surface) and two near the surface under a thin 10-15 cm layer of the mixture. Thin (0.5 cm diameter) plastic, 80 cm long tubes were connected to the funnels. A mobile gas analyser (GA2000) was used to measure the composition (by volume) of O2, CO2 and CH4 on a daily basis. The funnels were removed prior to each turning and reinserted afterwards. From each pair of funnels (core and surface) of both piles, one was kept closed between samplings. Two way ANOVA was used to test differences between piles and among the tubes. Post hoc Tukey tests were also used to further investigate these differences. There was a significant difference (at p<0.001) in the two piles for all three gases. The average concentrations of O2, CO2 and CH4 in Pile I, from all four funnels was 16.86%, 3.89% and 0.25%, respectively, where for Pile II the average values were 18.07%, 2.38% and 0.04%, respectively. The presence of OOMS in larger amounts in Pile I (resulting in more intense decomposing phenomena), and the larger particle size of OTB in Pile II (resulting in increasing porosity) are the probable causes of these significant differences. Samples from open funnels presented lower, but not significantly lower, O2 composition (higher for CO2 and CH4) in comparison with closed funnels in both depths and both piles. Not significant were also the different mean gas compositions between core and surface funnels in the same pile.     

Endpoint ‘floating leaves’ of <Emphasis Type="Italic">Potamogeton natans</Emphasis>: A new method to evaluate the development of macrophytes in pond mesocosms

Goal, Scope and Background One of the advantages of long-term mesocosm experiments as compared to short-term standard toxicity tests in the laboratory is the potential for detecting secondary effects due to the interaction of species and recovery with biomass of macrophytes being an important endpoint. However, generating biomass data by harvesting is often laborious, time-consuming, costly and restricted to the end of the experiment. Moreover, valuable information may get lost, in particular in single application studies, since maximal primary effects and secondary effects or recovery occur per se at different times. Potamogeton natans was used as an example in order to test whether number and area of floating leaves can be reliably measured and be used as intermediate and final endpoints in mesocosm effect studies. Methods Digital photos, which were taken of the water surface in the course of an indoor pond mesocosm study on herbicide effects, were subjected to image analysis. The results were compared to wet weight and ash-free dry weight of Potamogeton at the end of the herbicide study. Results and Discussion Both number and area of floating leaves indicated the same herbicide effects as wet weight and ash-free dry weight of Potamogeton. Error introduced by the different work steps is small and can be further minimised by a number of method improvements. Recommendations and Perspectives In indoor mesocosm studies, errors due to the perspective adjustment may be circumvented by taking the photos perpendicular to the water surface. Correction for lens aberration, identical light conditions and the use of fluorescence images are considered promising. Field applications are proposed.     

Tidal height,rather than habitat selection for conspecifics,controls settlement in mussels

Settlement is a major determinant of intertidal populations. However, the energy costs of lost larvae are very high. Accordingly, arrival and attachment on suitable substrata are essential requirements for species’ survival. On the intertidal, the presence of cues left by adult or juvenile conspecifics could be vital for the successful establishment of larvae arriving on the shore. Two mussel species, the indigenous Perna perna and the invasive Mytilus galloprovincialis, co-occur on the lower eulittoral zone on the south coast of South Africa. P. perna dominates the low and M. galloprovincialis the high mussel zones, with co-existence in the mid mussel zone. This study tested the hypothesis of settlement selectivity for conspecifics in these two mussel species, to understand whether the final adult distribution of mussels on the shores is determined by active behavioural and chemical mechanisms. Preferential selection by larvae for conspecifics was tested in the field during the peak settlement period in 2004 in natural mussel beds across zones and through manipulative experiments in the mid-zone where the species co-exist. On natural beds, settlement was determined by counts of settlers attached over 48 h onto artificial collectors. Collectors were placed on beds of P. perna and M. galloprovincialis present at both high- and low-adult densities, as well as in mixed beds. On such natural beds, settlers of both species consistently favored low-zone P. perna beds. Settlement patterns over 24 h onto experimentally created mussel patches consisting of P. perna, M. galloprovincialis or the two species combined beds, set in the mixed zone, did not conform with the results of the natural beds study: settlers of both species settled with no discrimination among different patches. The results indicate that mussels, which are sedentary, lack attraction to conspecifics at settlement. This highlights the importance of tidal height in setting settlement rates, and of post-settlement events in shaping populations of these broadcast spawners.     

Sustainable tourism development in remote regions? Questions arising from research in the North Kimberley,Australia

This paper is concerned with the sustainable development of a self-drive tourist destination in the remote region in Northern Australia. It first compares officially used data on visitor profiles to the information collected in the survey. Differences between data collected and available secondary data imply that tourism planning should include collection of region-specific tourism information to guide and mitigate potential environmental changes. Natural landscape features are the key attraction for visitors to the region. Visitor satisfaction levels with the current state of the destination are high. However, local residents reported perceived negative impacts of tourism, in particular on amenities and the ecology of the landscape. Visitors also reported behavior that might contribute to environmental impacts. Implications of the findings, in the context of future destination planning, are discussed.     

Determination of trace elements in aquatic plants by means of solid sampling Zeeman atomic absorption spectrometry (SS-ZAAS)

Direct solid sampling Zeeman atomic absorption spectrometric methods were developed and successfully applied to the determination of mercury, cadmium, lead and manganese in aquatic plants. Mercury was determined using a specially designed furnace. The other elements were determined by a third generation spectrometer. In 3-field- and dynamic mode the working range was substantially extended and high amounts of analyte were detectable without laborious dilution of solid samples. The measurements were based on calibrations using certified reference materials of organic origin. No matrix effects were observed under the optimised conditions. Results obtained were in good agreement with the certified values.