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.     

Long Term Trends in Concentration of Major Pollutants (SO<Subscript>2</Subscript>, CO,NO, NO<Subscript>2</Subscript>, O<Subscript>3</Subscript> and PM<Subscript>10</Subscript>) in Prague – Czech Republic (Analysis of Data Between 1992 and 2005)

To assess the effect of changes in traffic density and fuels used for heating at the beginning of the 1990s, 1992–2005 monthly averages of PM₁₀, SO₂, NO₂, NO, CO and O₃ from Prague, the Czech capital, were analyzed together with long term trends in emissions of major pollutants, fuel consumption and number of vehicles registered in Prague. The data from all monitoring stations were retrieved from the database of the state automated monitoring system. Correlation coefficients between ambient monthly averaged temperature and all pollutants of concern showed distinct seasonal trends. The results showed that while SO₂ and to some extent also CO concentrations dropped namely in the first half of the analyzed period (1992–1997) as a result decreased fossil fuel consumption for local heating, the behaviour of other pollutant concentrations followed a different pattern. PM₁₀ concentrations decreased during the beginning of the 1990s but showed a sign of increase after 2000. Concentrations of ozone and NO₂ did not reveal any significant change throughout the whole studied period. It can be concluded that during the studied period traditional urban sources of pollution, such as coal and oil combustion, lost their importance but were simultaneously substituted by pollutants from automotive transport (namely PM and NO₂) making the problem of air quality even worse.     

Observations of Atmospheric Nitrogen and Phosphorus Deposition During the Period of Algal Bloom Formation in Northern Lake Taihu,China

Cyanobacterial blooms in Lake Taihu occurred at the end of April 2007 and had crucial impacts on the livelihood of millions of people living there. Excessive nutrients may promote bloom formation. Atmospheric nitrogen (N) and phosphorus (P) deposition appears to play an important role in algal bloom formation. Bulk deposition and rain water samples were collected respectively from May 1 to November 30, 2007, the period of optimal algal growth, to measure the bulk atmospheric deposition rate, wet deposition rate, and dry deposition rate for total nitrogen (TN; i.e., all species of nitrogen), and total phosphorus (TP; i.e., all species of phosphorus), in northern Lake Taihu, China. The trends of the bulk atmospheric deposition rate for TN and the wet deposition rate for TN showed double peaks during the observation period and distinct influence with plum rains and typhoons. Meanwhile, monthly bulk atmospheric deposition rates for TP showed little influence of annual precipitation. However, excessive rain may lead to high atmospheric N and P deposition rates. In bulk deposition samples, the average percentage of total dissolved nitrogen accounting for TN was 91.2% and changed little with time. However, the average percentage of total dissolved phosphorus accounting for TP was 65.6% and changed substantially with time. Annual bulk atmospheric deposition rates of TN and TP during 2007 in Lake Taihu were estimated to be 2,976 and 84 kg km⁻² a⁻¹, respectively. The results showed decreases of 34.4% and 78.7%, respectively, compared to 2002–2003. Annual bulk deposition load of TN for Lake Taihu was estimated at 6,958 t a⁻¹ in 2007 including 4,642 t a⁻¹ of wet deposition, lower than the values obtained in 2002–2003. This may be due to measures taken to save energy and emission control regulations in the Yangtze River Delta. Nevertheless, high atmospheric N and P deposition loads helped support cyanobacterial blooms in northern Lake Taihu during summer and autumn, the period of favorable algal growth.     

An Assessment of Oil Pollution in the Coastal Zone of Patagonia,Argentina

The Patagonian coast is considered a relatively pristine environment. However, studies conducted along coastal Patagonia have showed hydrocarbon pollution mostly concentrated at ports that have fishing, oil loading, general merchant, and/or tourist activities. A high value of total aliphatic hydrocarbons (TAH) was found at the Rawson fishing port (741 μg/g dw). In other ports with and without petroleum-related activities, hydrocarbon values were approximately 100 μg/g dw. The highest values for TAH and total aromatic hydrocarbons (TArH) were found in Faro Aristizábal, north of San Jorge gulf (1304 and 737 μg/g dw, respectively). This is very likely the result of petroleum-related activities at the Comodoro Rivadavia, Caleta Cordova, and Caleta Olivia ports located within this gulf. In other coastal areas away from potential anthropogenic sources, hydrocarbon values were less than 2 and 3 μg/g dw for TAH and TArH, respectively. This review of published and unpublished information suggests that ports are important oil pollution sources in the Patagonian coast. More detailed studies are needed to evaluate the area affected by port activities, to understand the mechanisms of hydrocarbon distribution in surrounding environments, and to assess bioaccumulation in marine organisms. Despite that some regulations exist to control oil pollution derived from port and docked vessel activities, new and stricter management guidelines should be implemented.     

Assessing Landowner Activities Related to Birds Across Rural-to-Urban Landscapes

Fluctuations of bird abundances in the Midwest region of the United States have been attributed to such factors as landscape change, habitat fragmentation, depredation, and supplemental feeding. However, no attempt has been made to estimate the collective role of landowner activities that may influence birds across a landscape. To investigate how landowners might influence birds when the majority (> 90%) of land is privately owned, we surveyed all 1694 private domestic landowners living on three breeding bird survey routes (120 km) that represent a continuum of rural-to-urban landscapes in Southeastern Michigan from October through December 2000. Our survey was designed to investigate (1) the proportion of landowners involved in bird feeding, providing bird houses, planting or maintaining vegetation for birds, gardening, landscaping, applying fertilizer, and applying pesticides or herbicides; (2) whether differences existed between urban, suburban, and rural landowner activities; and (3) whether landowners that carried out a given activity were sociodemographically different from those who did not. Of the 968 respondents (58.5% response rate), 912 (94%) carried out at least one of the activities on their land and the average landowner carried out 3.7 activities. A total of 65.6% fed birds, 45.7% provided bird houses, 54.6% planted or maintained vegetation for birds, 72.7% gardened, 72.3% landscaped, 49.3% applied fertilizer, and 25.2% applied pesticides or herbicides. Significant differences existed between the landscapes, with rural landowners having more bird houses and applying pesticides or herbicides in greater frequency. Similarly, urban landowners had a greater density of bird feeders and houses, but planted or maintained vegetation in the lowest frequency. Participation in activities varied by demographic factors, such as age, gender, and occupation. Scaling each activity to all landowners, including nonrespondents, across all landscapes indicates that between 14% and 82% of landowners may be engaged in a particular activity, with application of pesticides or herbicides having the least potential involvement (13.9%–55.4%) and gardening having the greatest potential involvement (40.1%–81.6%). Taken collectively, our results indicate that landowners are both intentionally and unintentionally engaged in a wide range of activities that are likely to influence bird populations.     

Nutrient-impregnated charcoal: an environmentally friendly slow-release fertilizer

The widespread contamination of surface and ground water quality from the heavy use of fertilizer in modern agriculture is the current concern. Therefore, this study was carried out to develop a slow-release fertilizer using charcoal. The morphology of the charcoal impregnated fertilizer was investigated by scanning electron microscopy (SEM). This study also evaluated the release patterns of N, P, and K from impregnated charcoal using a simulated soil solution and distilled water as leaching solutions. The patterns of N, P, and K releases were examined in both static and continuous-flow conditions for 360 h. Releases of N, P, and K from impregnated charcoal were found to be slow and steady. However, the release trends of N, P, and K were higher in soil solution than distilled water under both the above conditions. Dissolution occurred when N, P, and K were released in the above leached solutions. As a result, the fertilizer impregnated charcoal could be developed as slow-release type fertilizer to minimize the contamination.     

Phosphates for Pb immobilization in soils: a review

In its soluble ionic forms, lead (Pb) is a toxic element occurring in waters and soils mainly as the result of human activities. The bioavailability of lead ions can be decreased by complexation with various materials in order to decrease their toxicity. Pb chemical immobilization using phosphate addition is a widely accepted technique to immobilize Pb from aqueous solution and contaminated soils. The application of different P amendments cause Pb in soils to shift from forms with high availability to the most strongly bound Pb fractions. The increase of Pb in the residual or insoluble fraction results from formation of pyromorphite Pb₅(PO₄)₃X where X = F, Cl, Br, OH, the most stable environmental Pb compounds under a wide range of pH and Eh natural conditions. Accidental pyromorphite ingestion does not yield bioavailable lead, because pyromorphite is insoluble in the intestinal tract. Numerous natural and synthetic phosphates materials have been used to immobilize Pb: apatite and hydroxyapatite, biological apatite, rock phosphate, soluble phosphate fertilizers such as monoammonium phosphate, diammonium phosphate, phosphoric acid, biosolids rich in P, phosphatic clay and mixtures. The identification of pyromorphite in phosphate amended soils has been carried out by different non destructive techniques such as X-ray diffraction, scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, X-ray absorption fine structure, transmission electron microscopy and electron microprobe analysis. The effectiveness of in situ Pb immobilization has also been evaluated by selective sequential extraction, by the toxicity leaching procedure and by a physiologically based extraction procedure simulating metal ingestion and gastrointestinal bioavailability to humans. Efficient Pb immobilization using P amendments requires increasing the solubility of the phosphate phase and of the Pb species phase by inducing acid conditions. Although phosphorus addition seems to be highly effective, excess P in soil and its potential effect on eutrophication of surface water, and the possibility of As enhanced leaching remains a concern. The use of mixed treatments may be a useful strategy to improve their effectiveness in reducing lead phyto- and bioavailability.     

Report on the fifth meeting of INRA’s national network of ecotoxicologists,ECOTOX

The fifth meeting of INRA’s national network of ecotoxicologists took place on 25 to 27 November 2014 in Biarritz, France. The main aim of the meeting was to bring together ecotoxicologists from INRA and associated partners, providing them ample opportunity to share and discuss their latest scientific results as well as the national policy of research in ecotoxicology and to precise perspectives for the network.