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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.

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.

The ability of biomarkers to integrate effects of chemicals on biota has lead to increased calls for their application in assessing the status of polluted ecosystems. In tandem there has been an increase in our knowledge of the ecophysiological responses of keystone species to pollutants, which has allowed the development of a number of promising methods. In contrast to the number of biomarker development studies, the number of biomarker validation studies has remained limited. This paper redresses this imbalance by drawing together data from studies that have used the earthworm lysosomal membrane stability response (measured using the neutral red retention assay). This review first gives a short history of the biomarker's development. Second, it sets published applications of the technique against established criteria for a "good" biomarker (i.e., dose-response relationship, sensitivity, ecological relevance, confounding factors, chemical specificity, species differences, time-response relationship, methodological concerns, and overall public/regulator confidence, and acceptance). Discussion of the biomarker's suitability to each criterion is followed by an overall evaluation of its workability for routine soil quality assessment and caveats for its use.