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David J Spurgeon

Publications and source records attributed to David J Spurgeon.

18 recordsLinked to original sources

Exploiting Omic Data to Advance Predictive Ecotoxicology.

Predicting species-specific chemical sensitivity using in silico approaches has the potential to transform environmental risk assessment, conservation, and biomonitoring, while reducing, and ultimately replacing, animal testing. Genomic and transcriptomic data capture extensive sensitivity-relevant variation, including differences in molecular targets, xenobiotic metabolism, and damage mitigation pathways. Large-scale sequencing initiatives therefore offer an unprecedented opportunity to address ecotoxicology's "too many species" problem. Although existing omic-based predictive tools provide proof of concept, they have so far been applied to a narrow set of relatively straightforward prediction scenarios. To achieve broader applicability, current and future tools must be firmly grounded in the diverse molecular mechanisms underlying differential chemical responses. Here, we critically evaluate the emerging field of predicting species sensitivity using molecular variation inferred from omic data. We analyze the strengths and limitations of current omic-based approaches and identify major sequence and ecotoxicological data gaps, as well as critical bioinformatic challenges. We then review the current knowledge of how molecular biology underlies differential chemical sensitivity, outlining research paths to allow the next generation of sensitivity prediction tools to exploit ever expanding omic data.

Ecotoxicology↗

Effect of temperature and season on reproduction, neutral red retention and metallothionein responses of earthworms exposed to metals in field soils.

This study investigated the short-term survival, reproduction and physiological (lysosomal membrane stability, metallothionein transcript copy number, body tissue metal concentrations) responses of Lumbricus rubellus exposed to metal contaminated field soils under different laboratory temperatures (10, 15 and 20 degrees C) and physiological responses of earthworms collected from the field in three different seasons (spring, autumn, winter). In the laboratory, metal contaminated soils had significant effects on reproduction (p<0.001), metallothionein-2 (MT-2) expression (p=0.033) and earthworm As (p=0.003), Cd (p=0.001), Pb (p<0.001) and Zn (p<0.001) concentration, but not lysosomal membrane stability and tissue Hg and Cu. No effect of temperature was found for any parameter. Principal component analysis of extractable and tissue metal concentrations indicated PC1 as a measure of metal stress. Both cocoon production (r=-0.75) and MT-2 induction (r=0.41) were correlated with PC1. A correlation was also found between cocoon production and MT-2 expression (r=-0.41). Neutral red retention and MT-2 measurements in worms collected from the field sites in three seasons confirmed the absence of a temperature effect on these responses.

Animals↗

Developing a critical load approach for national risk assessments of atmospheric metal deposition.

The critical load approach has been proposed for evaluation of the need to reduce atmospheric emissions of metals that lead to transboundary transport and deposition across Europe. The present study demonstrates and evaluates the application of a critical load approach for national-scale risk assessment of metal deposition in the United Kingdom. Critical load maps, calculated using critical limits based on pH-dependent free metal ion activities, are presented. Current concentrations of lead and cadmium in soils are compared with two sets of critical limit values: First, limits based on the reactive soil concentration, and second, a pH-dependent free ion critical limit function, which takes into account variable soil characteristics across the country. The use of these two models leads to different conclusions about which areas of the United Kingdom are at greatest risk, partly because of differences in the range of values of pH and organic matter in soils used in ecotoxicological experiments and in the national database. Critical loads were calculated based on free ion critical limits; the critical loads were lowest in the south and east of the country and were associated with higher soil pH, lower runoff, and lower soil organic matter.

Atmosphere↗

Effect of pH on metal speciation and resulting metal uptake and toxicity for earthworms.

In the present study, relationships between changes in the solubility and speciation of metals in contaminated soils under different pH regimes and their toxicity to earthworms were investigated. Earthworms (Lumbricus rubellus) were exposed in a laboratory bioassay to metalliferous soils under three pH regimes: Unamended pH, pH lowered by one unit (pH -1), and pH increased by one unit (pH +1). In each soil, total (hot nitric acid-extractable) and 0.01 M CaCl2-extractable metal concentrations were measured and soil pore-water chemistry analyzed to allow metal speciation to be modeled using the Windermere Humic Aqueous Model. Earthworm metal accumulation was determined and toxicity assessed by measuring survival and reproduction and at the molecular level by recording expression of the gene encoding metallothionein-2 (MT-2) using quantitative reverse transcriptase-polymerase chain reaction. Both metal solubility and speciation were found to be highly pH dependent. Metal accumulation in earthworms was influenced by soil concentration and, in some cases (e.g., Cd), by pH. Reproduction was affected (reduced up to 90%) by soil metal level, pH, and their interaction. Relationships between analyzed and calculated Zn concentrations and toxicity and between analyzed and calculated Cd concentrations and tissue accumulation and MT-2 expression were compared by fitting dose-response models and assessing the fit of the data. This analysis indicated that values based on a pH-adjusted free ion concentration best explained toxicity (r2 = 0.82) and accumulation (r2 = 0.54). Expression of MT-2 was, however, poorly correlated (p > 0.05) with all analyzed and modeled soil metal concentrations.

Animals↗

Hierarchical responses of soil invertebrates (earthworms) to toxic metal stress.

The concept of a hierarchical cascade of biological responses to stress occurring across different levels of biological organization is an underlying principle of both theoretical and regulatory ecology/ecotoxicology. This study investigates the reality of this cascade for earthworms exposed to toxic metal stress. Gene expression was the most sensitive endpoint (EC50 = 616 microg Zn g(-1)) followed by the integrity of coelomocyte lysosomal membrane (EC50 = 645 microg Zn g(-1)). This confirms that, in accordance with the cascade concept, suborganism level endpoints respond at lower metal concentrations than higher organization endpoints. The relative sensitivity of the higher organization parameters was not as predicted by the cascade. Organic material removal was more sensitive (EC50 = 997 microg Zn g(-1)) than L. rubellus reproduction (EC50 = 3236 microg Zn g(-1)), L. rubellus population size (EC50 = 5000-11500 microg Zn g(-1)), and earthworm community diversity (EC50 = 1737 microg Zn g(-1)). This can be attributed to (1) the relative insensitivity of L. rubellus to metals and (2) general toxic effects of metals on the earthworm energy budget (and thus feeding). On the basis of these results, it can be concluded that predictive assessments of the consequences of environmental stressors needs to include approaches that respect the relative sensitivities of different taxa, while retrospective appraisals should exploit the sensitivity of low organization level responses.

Animals↗

Establishing principal soil quality parameters influencing earthworms in urban soils using bioassays.

Potential contamination at ex-industrial sites means that, prior to change of use, it will be necessary to quantify the extent of risks to potential receptors. To assess ecological hazards, it is often suggested to use biological assessment to augment chemical analyses. Here we investigate the potential of a commonly recommended bioassay, the earthworm reproduction test, to assess the status of urban contaminated soils. Sample points at all study sites had contaminant concentrations above the Dutch soil criteria Target Values. In some cases, the relevant Intervention Values were exceeded. Earthworm survival at most points was high, but reproduction differed significantly in soil from separate patches on the same site. When the interrelationships between soil parameters and reproduction were studied, it was not possible to create a good model of site soil toxicity based on single or even multiple chemical measurements of the soils. We thus conclude that chemical analysis alone is not sufficient to characterize soil quality and confirms the value of biological assays for risk assessment of potentially contaminated soils.

Animals↗

Earthworm responses to Cd and Cu under fluctuating environmental conditions: a comparison with results from laboratory exposures.

Laboratory toxicity tests are usually conducted under stable ambient conditions, while exposures in ecosystems occur in a fluctuating climate. To assess how climate influences the toxicity of Cu and Cd for the earthworm Lumbricus rubellus, this study compared effects for life-cycle parameters (survival, reproduction), cellular status (lysosomal membrane stability), gene expression (transcript of the metal binding protein metallothionein-2) and tissue metal concentration measured under outdoor conditions, with the same responses under constant conditions as measured by Spurgeon et al. [Spurgeon, D.J., Svendsen, C., Weeks, J.M., Hankard, P.K., Stubberud, H.E., Kammenga, J.E., 2003. Quantifying copper and cadmium impacts on intrinsic rate of population increase in the terrestrial oligochaete Lumbricus rubellus. Environmental Toxicology and Chemistry 22, 1465-1472]. Both metals were found to significantly influence earthworm reproduction, compromise lysosomal membrane stability and induce MT-2 gene expression in the outdoor system. Comparison with physiological and life-cycle responses in the laboratory indicated similar response patterns and effect concentrations for Cu. For Cd, lysosomal membrane stability and MT-2 expression showed comparable responses in both exposures. Juvenile production rate, however, gave different dose response relationships, with the EC-(50) in the outdoor test approximately half that in the laboratory test. A difference in Cd accumulation was also seen. Overall, however, the comparison indicated only a marginal effect of environmental fluctuations typical for northern temperate Europe on earthworm sensitivity to the two metals.

Animals↗

Biological assessment of contaminated land using earthworm biomarkers in support of chemical analysis.

Biological indicators can be used to assess polluted sites but their success depends on the availability of suitable assays. The aim of this study was to investigate the performance of two earthworm biomarkers, lysosomal membrane stability measured using the neutral red retention assay (NRR-T) and the total immune activity (TIA) assay, that have previously been established as responsive to chemical exposure. Responses of the two assays were measured following in situ exposure to complexly contaminated field soils at three industrial sites as well as urban and rural controls. The industrial sites were contaminated with a range of metal (cadmium, copper, lead, zinc, nickel and cobalt) and organic (including polycyclic aromatic hydrocarbons) contaminants, but at concentrations below the 'New Dutch List' Intervention concentrations. Exposed earthworms accumulated both metals and organic compounds at the contaminated sites, indicating that there was significant exposure. No effect on earthworm survival was found at any of the sites. Biomarker measurements, however, indicated significant effects, with lower NRR-T and TIA found in the contaminated soils when compared to the two controls. The results demonstrate that a comparison of soil pollutant concentrations with guideline values would not have unequivocally identified chemical exposure and toxic effect for soil organisms living in these soils. However, the earthworm biomarkers successfully identified significant exposure and biological effects caused by the mixture of chemicals present.

Animals↗

Deriving soil critical limits for Cu, Zn, Cd, and Pb: a method based on free ion concentrations.

We present a method to calculate critical limits of cationic heavy metals accounting for variations in soil chemistry. We assume the free metal ion concentration (Mfree) to be the most appropriate indicator of toxicity, combined with a protective effect of soil cations (e.g., H+, Ca2+). Because soil metal cations tend to covary with pH, the concentration of Mfree exerting a given level of toxic effect (Mfree,toxic) can be expressed as a function of pH alone. We use linear regression equations to derive Mfree,toxic in toxicity experiments from soil pH, organic matter content, and endpoint soil metal. Chronic toxicity data from the literature, for plants, invertebrates, microbial processes, and fungi are interpreted in terms of an average log Mfree,toxic together with distributions of species sensitivity. This leads to critical limit functions to protect 95% of species, of the form log Mfree,CRIT = (pH + gamma. Appreciable effects of soil pH upon log Mfree,CRIT are found, with alpha = -1.21 (Cu), -0.34 (Zn), -0.43 (Cd), and -0.83 (Pb). Critical limit functions in terms of the geochemically active soil metal (Msoil,CRIT), that pool of metal which controls the free ion concentration, have also been derived, with soil pH and organic matter content as variables. The pH effect on Msoil,CRIT is relatively small, with slopes of 0.05 (Cu), 0.19 (Zn), 0.16 (Cd), and 0.20 (Pb), since the effect of pH on Mfree,CRIT is countered by the variation of Mfree with pH.

Cations↗

Pedological characterisation of sites along a transect from a primary cadmium/lead/zinc smelting works.

A pedological characterisation of seven sites along a transect from a smelter at Avonmouth, UK, was undertaken. Site locations comprised a mixture of both grassland (5 sites) and oak tree dominated (2 sites) areas. Geographically, sites were either low lying or on adjacent elevated ground. Across the transect, a severe gradient of metal concentrations was found with highest values for organic soil horizons from close to the factory. Comparisons with quality standards indicate that these metal levels are likely to be a major ecological stressor. In addition to the strong metal gradient, a number of other between site differences were also observed. These were firstly water regime, which at the low lying sites close to the smelter showed influence by both, groundwater and stagnant water, while at more distant sites indicated susceptibility to stagnant water only, and secondly soil pH, which showed large between site variations, although no consistent trend along the transect. Humus forms at sites more than 1.5 km from the smelter were characteristic for the soil conditions and land-use present, while those at sites within 1.5 km showed disturbed profiles. Indeed, the humus types present at these locations suggest that the activity of soil invertebrates may be inhibited. This is almost certainly due primarily to the high concentrations of metals that were present in mineral soil and organic horizons at the sites. However, the potential influences of water regime and pH could also be relevant factors influencing the results of any further studies conducted at these study sites.

Cadmium↗

Environmental metabonomics: applying combination biomarker analysis in earthworms at a metal contaminated site.

Earthworms were taken across an environmental gradient of metal contamination for ecotoxicology assessment. Both indigenous (Lumbricus rubellus and L. terrestris) and introduced earthworms (Eisenia andrei, exposed in mesh bags) were studied. Changes in the levels of small molecule metabolites in earthworm tissue extracts were analysed by 1H NMR spectroscopy as a means of identifying combination biomarker compounds. Principal components analysis of the NMR spectral data revealed that biochemical changes were induced across the metal contamination gradient. Native worms (L. rubellus) from the most polluted sites were associated with an increase in the relative concentration of maltose; a decrease was also seen in the concentration of an as yet unidentified biomarker compound. Introduced worms (E. andrei) did not show differences to the same extent. Direct integration of the resonances from histidine and 1-methylhistidine showed that relative histidine concentrations were elevated slightly for L. rubellus, confirming the results of earlier mesocosm studies. Conversely, the relative concentrations of both histidine and 1-methylhistidine were greatly reduced by metal contamination in L. terrestris. This study demonstrates the utility of NMR spectroscopy in detecting previously unknown potential biomarkers for ecotoxicity testing and identified maltose as a potential biomarker compound deserving of further study.

Animals↗

Metal effects on soil invertebrate feeding: measurements using the bait lamina method.

Heightened concerns regarding the protection of terrestrial ecosystems at a national level has increased the need to develop a suite of indicators capable of assessing the quality, integrity and fertility of soils. Of the assays currently available, tests that measure aspects of soil function and associated parameters are among the most promising, since these integrate effects on soil quality at the highest level of organisation. In this study we describe results of the deployment of an indicator of soil functional integrity (the bait lamina test) that is designed to measure the feeding activity of soil invertebrates. Bait lamina was used at six grassland sites located along a transect from a smelter at Avonmouth (South-West England) used in the EU funded BIOPRINT II project. Results indicated highest bait removal (feeding) at sites furthest from the factory, intermediate feeding activity at intervening sites and extremely low activities at the two sites closest to the smelter. The strong decline in activity for the group of sites closest to the smelter corresponded with increasing metal concentrations suggesting a clear impact of metals on detritivorous invertebrate feeding. Comparisons of the results of the bait lamina study to previous invertebrate survey work suggested that the differences in observed bait removal can be attributed to direct effects of metals on the abundance and biodiversity of key decomposer groups such as earthworms, isopods, molluscs, myriapods, springtails and mites.

Animals↗

Critical analysis of soil invertebrate biomarkers: a field case study in Avonmouth, UK.

During the period 1996-1999 a joint field research programme (BIOPRINT-II) funded by the European Union was undertaken. The main objective of this project was the deployment of biochemical fingerprint techniques of soil invertebrate biomarkers for assessing the exposure and effect of toxicants on soil invertebrates in the field. The aim was to apply these techniques in the field focusing on a a chronically polluted field near a lead and zinc smelter in Avonmouth (UK). Therefore six sites were selected from which organisms were either sampled or transplanted to or from the laboratory. The project has provided a unique opportunity to apply a series of biological test methodologies in order to determine the hazard posed to soil sustainability and by inference soil biodiversity and function. This work has attempted to understand the linkage between effects measured at the molecular or cellular level and relate these to changes at higher levels of biological organisation. Here we evaluated the links between biomarkers and soil function parameters. The paper aims to summarize and explore the necessary caveats that must be understood before soil biomarker test systems may be used to strengthen the risk assessment process.

Animals↗

Toxicological, cellular and gene expression responses in earthworms exposed to copper and cadmium.

This study correlates sub-organismal changes with toxicological effects in earthworms (Lumbricus rubellus) exposed to copper and cadmium. Both metals reduced survival and reproduction at the highest concentration (LC50 5.11 microM Cu g(-1) and 4.04 microM Cd g(-1); cocoon production EC50s 5.17 microM Cu g(-1) and 1.86 microM Cd g(-1), all values as dry mass soil). Cadmium significantly reduced lysosomal membrane stability (at 1.86 microM Cd g(-1) and higher), upregulated metallothionein gene expression (at least sevenfold in all treatments) and reduced lysosome-associated-glycoprotein gene expression. Copper did not lower lysosomal membrane stability, but did upregulate metallothionein gene expression (at 2.5 microM Cu g(-1)), reduce lysosome-associated-glycoprotein gene expression and gave a nonlinear pattern for mitochondrial ribosomal subunit transcript expression (reduced at 0.35 and 0.811 microM Cu g(-1); higher at 2.5 microM Cu g(-1)). Correlation of metal body residue concentrations and cellular and molecular genetic responses with juvenile production rate confirmed a relationship for metallothionein expression, lysosomal membrane stability and cadmium tissue concentration in cadmium-exposed worms. Relationships between responses were also found for both metals. These suggested mechanisms for the interaction of cadmium and copper with specific gene products and with organelle (mitochondrial, lysosomal) functioning.

Animals↗

Comparison of instantaneous rate of population increase and critical-effect estimates in Folsomia candida exposed to four toxicants.

The instantaneous rate of population increase (ri) integrates several life cycle variables into one accessible statistic and has been proposed as a more practical alternative than assembling full life tables in the study of population-level responses to toxicant exposure. In this study the sensitivity of instantaneous rate of population increase is compared to critical-effect estimates for populations exposed to four toxicants with different modes of action. Populations of the Collembolan Folsomia candida were exposed to cadmium, copper, pyrene, and chlorpyrifos in artificial soil following the standardized ISO (International Organization for Standardization, Geneva, 1999) protocol. We calculated ri values and LC(50), EC(50juvenile), and NOEC values for each chemical. Comparison of the relative toxicity of the four chemicals indicated that chlorpyrifos had the lowest values and was thus the most toxic, followed by pyrene, cadmium, and copper. Significant changes in ri were seen to follow closely changes in the sublethal parameter measured (juvenile production) and showed populations in decline at concentrations as low as 40% of the LC(50). The study showed ri to be a good measure of population response, and we conclude that the statistic gives a better understanding of effects on a population than through the sole use of traditional critical-effect estimates.

Animals↗

Closing the loop: a spatial analysis to link observed environmental damage to predicted heavy metal emissions.

In many cases, the link between industrial emissions and damage to the environment can only be inferred. The Environment Agency of the United Kingdom imposes emissions limits on industrial sites so that predicted concentrations and deposition rates remain below standard thresholds. Estimates of appropriate critical levels and loads are usually based on laboratory results and rarely estimate synergistic effects between pollutants or consider biological adaptation or selection in the target receptor organisms. The Avonmouth smelter has been emitting zinc and other heavy metals since 1929. It has been the subject of a number of detailed and synoptic studies, especially the impact on soil invertebrates. Damage was assessed using both physiological and ecological measurements. Two methods of spatial analysis were investigated, namely interpolation using standard geographical information system (GIS) operators and atmospheric dispersal modeling using an off-the-shelf model. Both methods can be used to compute contours (isolines) of predicted biological effect. Correlation results show that dispersal modeling is at least as good as kriging but requires much less data. This article demonstrates the usefulness of GIS and dispersal models as tools in decision making to determine the most suitable sampling sites in the assessment and monitoring of the impact of contamination around major point sources.

Air Pollution↗

Quantifying copper and cadmium impacts on intrinsic rate of population increase in the terrestrial oligochaete Lumbricus rubellus.

Demographic methods can translate toxicant effects on individuals into consequences for populations. To date few such studies have been conducted with longer-lived invertebrates. This is because full life-cycle experiments are difficult with such species. Here we report the effects of copper and cadmium on the key demographic parameter intrinsic rate of population increase (r) in a long-lived invertebrate (an earthworm). The approach used to derive r was based on robust measurement of effects on life-cycle traits in three specifically designed toxicity tests and integration of this data within a demographic model. The three laboratory tests used to generate values for specific life-cycle parameters under copper and cadmium exposure were suited to the task. Significant effects on a range of separate adult and juvenile life-cycle parameters were seen. Integration of parameter values within the demographic equation indicated that for copper, r was reduced only at a concentration that also caused adult mortality. For cadmium, a more graded exposure-dependent effect on fitness was seen, with r reduced at sublethal concentrations. The concentration response patterns for r found for the two metals suggest significantly different consequences for earthworm populations exposed to sublethal levels of copper and cadmium.

Animals↗

Earthworm species of the genus Eisenia can be phenotypically differentiated by metabolic profiling.

The universality of low molecular weight metabolites allows rapid and straightforward investigation of the biochemistry of genetically uncharacterised species. Thus ex vivo metabolic profiling in combination with multivariate data analysis (metabonomics) offers great potential in comparative biology. Here we present the first use of high resolution nuclear magnetic resonance (NMR) spectroscopy to distinguish closely related animal species via their metabolic phenotype (metabotype). We have profiled the three Eisenia (Oligochaeta, Lumbricidae) species Eisenia fetida, Eisenia andrei and Eisenia veneta using tissue extracts and coelomic fluid analysis. The low molecular weight biochemical profiles of tissue extracts were highly conserved for all three species, with E. fetida and E. andrei being more similar to each other than to E. veneta. However the metabolic profiles of the coelomic fluid of the different species were highly distinctive - the NMR spectra allowed unequivocal identification of species. Multivariate statistics were also used to quantify these spectral differences and to enable simplified graphical visualisation of species similarity. These results show that two morphologically undistinguishable species (E. fetida and E. andrei) differ markedly in their biochemical profiles despite apparently occupying the same ecological niche, and indicate that metabolic phenotype profiling can be used as a powerful functional genomics tool.

Animals↗