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Ring-testing and field-validation of a terrestrial model ecosystem (TME)--an instrument for testing potentially harmful substances: effects of carbendazim on earthworms.

The effects of the fungicide carbendazim (applied in the formulation Derosal) on earthworms (Lumbricidae) was determined in Terrestrial Model Ecosystem (TME) tests and field-validation studies. TMEs consisted of intact soil columns (diameter 17.5 cm; length 40 cm) taken from a grassland or, in one case, from an arable site. The TMEs were taken from the same site where the respective field-validation study was performed. The tests were performed in Amsterdam (The Netherlands), Bangor (Wales, UK), Coimbra (Portugal) and Flörsheim (Germany). The sites selected had an earthworm coenosis representative of the different land use types and regions. In addition, the differences between the coenosis found in the TMEs and the respective field sites were in general low. A high variability was found between the replicate samples, which reduces the probability of determining significant differences by the statistical evaluation of the data. Similar effects of the chemical treatment were observed on abundance as well as on biomass. Effects were most pronounced 16 weeks after application of the test chemical. The observed effects on earthworm abundance and biomass did not differ between the TME tests and the respective field-validation studies. Effects on earthworm diversity were difficult to assess since the number of individuals per species was low in general. However, the genus Lumbricus and in particular L. terrestris and L. rubellus seemed to be more affected by the chemical treatment than others. NOEC and EC50-values derived from the TME pre-test, the TME ring-test and the field-validation study indicate that the TMEs of the different partners delivered comparable results although different soils were used. Due to the high variability NOECs could often not be determined. The EC50-values for the effect of carbendazim on earthworm abundance ranged between 2.04 and 48.8 kg a.i./ha (2.71-65.2 mg/kg soil) and on earthworm biomass from 1.02 to 34.6 kg a.i./ha (1.36-46.0 mg/kg soil). These results indicate that the abundance and biomass of earthworms are suitable endpoints in ecotoxicological studies with TMEs.

Animals↗

Single and joint effects of acetochlor and urea on earthworm Esisenia foelide populations in phaiozem.

Much attention is paid to soil health and environmental safety. Earthworms are an important indicator of soil ecosystem health and safety. Ecological toxicity of acetochlor and excessive urea, in both their single and joint effects, on earthworm Esisenia foelide was thus studied using the soil-culture method. Acetochlor had an enhanced toxicity from low concentration to high concentration. The mortality of earthworms after a 6-day exposure was changed from 0 to 86.7%, and the weight change rate ranged from 7.86 to -30.43%, when the concentration of acetochlor was increased from 164 to 730 mg kg(-1). Urea expressed its positive and beneficial effects on earthworms when its concentration was lower than 500 mg kg(-1). Strongly toxic effects took place when the concentration of urea was higher than 1000 mg kg(-1). The mortality of earthworms exposed to urea reached 100% when its concentration was more than 1500 mg kg(-1). When the concentration of urea was lower than 500 mg kg(-1), there were antagonistic effects between the two agrochemicals on earthworms; when the concentration of urea was higher than 500 mg kg(-1), joint toxic effects of acetochlor and excessive urea on earthworms were synergic. In any case, excessive urea application is very harmful to the health of soil ecosystems.

Animals↗

The effect of lead-contaminated soil from Canadian prairie skeet ranges on the neutral red retention assay and fecundity in the earthworm Eisenia fetida.

The sublethal effects of lead (Pb) on the earthworm Eisenia fetida were evaluated in the laboratory using freshly spiked soil and soil collected from Canadian prairie skeet ranges. After a four-week exposure to soil spiked with lead acetate, earthworm neutral red retention time (NRRT). soil Pb concentrations, and earthworm Pb body burdens were measured. Lysosomal NRRT was reduced in a concentration-dependent manner (p < 0.0001), and NRRT was negatively correlated with earthworm Pb body burdens (r = -0.80, p < 0.0001). To evaluate the effects of aged Pb, earthworms were exposed to soil from three skeet ranges, and responses were compared with three matched reference sites. After a four-week exposure, NRRT, growth, fecundity, soil total Pb levels, and earthworm Pb body burdens were measured. The potentially bioavailable fraction of Pb in these sites was measured using a Ca(NO3)2 extraction. Growth and fecundity did not differ significantly between any of the skeet ranges and their reference sites. However, NRRT was significantly reduced in all three ranges compared with their respective reference sites (p < 0.05), indicating that the neutral red retention assay (NRRA) may be useful for detecting toxicity and potential hazards at Pb-contaminated sites. Lysosomal NRRT was negatively correlated with soil Ca(NO3)2-extractable Pb (r = -0.80, p < 0.0001) and soil total Pb (r = -0.73, p = 0.001). Lysosomal NRRT was negatively correlated (r = -0.67, p < 0.002) with earthworm Pb tissue levels.

Animals↗

[Acute toxicological effects of heavy metal pollution in soils on earthworms].

Acute and sub-acute lethal effects of single and combined Cu, Zn, Pb and Cd on earthworm was determined using meadow brown soil. Results indicated that the mortality of earthworm was significantly related with the concentrations of Cu and Zn in the soil (alpha = 0.05, RCu = 0.86, RPb = 0.87), and the inhibition rate of earthworm growth was significantly related with the concentration of Cu in the soil (alpha = 0.05, RCu = 0.84). There was no significant relation between the concentrations of other heavy metals tested and the death rate and the growth-inhibition rate. The degree of individual earthworm enduring the toxicity of heavy metals varied greatly. The threshold concentration of toxicity defined by statistical method, the level to conduce the death of individual earthworm was 300 mg.kg-1 for Cu, 1300 mg.kg-1 for Zn, 1700 mg.kg-1 for Pb, 300 mg.kg-1 for Cd. LC50 was 400-450 mg.kg-1 for Cu, 1500-1900 mg.kg-1 for Zn, 2350-2400 mg.kg-1 for Pb and 900 mg.kg-1 for Cd. Combined effects of single Cu, Zn, Pb and Cd to conduce more than 10% of the death rate of earthworm could result in 100% of the death rate of earthworm, thus showing the strong synergistic joint effect of the heavy metals.

Animals↗

[Effects of different vegetation restoration of degraded red soil on earthworm population dynamics].

This study was conducted at the long-term experimental plots in Ecological Experimental Station of Red Soil in Yujiang County (28 degrees 15'30''N, 116 degrees 55'30''E ), Jiangxi Province, subtropical China. Earthworm population was investigated seasonally from May 1999 to February 2000, under different vegetations including four artificial woodlands [deciduous broadleaf woodland (Quercus chenii, Qc), evergreen broadleaf woodland (Schima superba, Ss), coniferous woodland (Pinus massonina, Pm) and mixed woodland (Schima superba-Pinus massonina, Sm)], two grasslands [gently-disturbed grassland (G1), undisturbed grassland (G2)] and control wasteland (CK). The results indicated that the population structure was very simple. Only Drawinda gisti characterized by pioneer was found. The seasonal averages of density and biomass were in the order of G2 > G1 > Qc > Ss > Pm > Sm > CK, and those of G2, G1 and Qc were significantly higher than those of the latters (P < 0.05). Seasonal fluctuations were obvious with dry-hot summer depressing the earthworm population sharply, leading to the aestivation of earthworm. Based on the variation coefficients of density and biomass, Qc had the highest ecosystem stability, followed by Sm and Ss, and G1, G2, and Pm had the lowest stability. The overall differentiation of earthworm population could be drawn through canonical discriminant analysis. There were significant correlations between earthworms and some soil properties (P < 0.01). Overall, the differentiation of earthworm population was driven by the quantity and quality of soil organic matter returned by the vegetations. Additionally, based on earthworm population, the importance of selecting appropriate vegetation types during the restoration of degraded red soil was emphasized.

Animals↗

Toxic effects of acetochlor and methamidophos on earthworm Eisenia fetida in phaiozem, northeast China.

Acetochlor and methamidophos are two important agrochemicals which are widely applied to agricultural production in northeast China. The investigation on the earthworm Eisenia fetida as an important type of soil animals exposed to single and binary-combined contamination of acetochlor and methamidophos was thus carried out. The single toxic effect test showed that the two agrochemicals had their toxicity to the earthworms living in phaiozem. Acetochlor had a stronger acute toxic effect on the earthworms than methamidophos. The mortality of the earthworms exposed to individual acetochlor and methamidophos changed with an increase in the exposure time and the exposed concentrations. The LD50 value of acetochlor and methamidophos toxic to the earthworms was 115.6-275.3 and 29.5-228.6 mg/kg, respectively. The weight of the earthworms was a more sensitive index compared to the mortality in indicating toxic effects of acetochlor and methamidophos in phaiozem. When considering both the mortality and the body-weight change, the combined pollution of acetochlor and methamidophos in phaiozem resulted in their synergic toxic effects on the earthworms.

Animals↗

Immunocytochemical electron microscopic study and Western blot analysis of troponin in striated muscle of the fruit fly Drosophila melanogaster and in several muscle cell types of the earthworm Eisenia foetida.

BACKGROUND: There is little information about troponin in invertebrate muscles, and no previous references to this protein in annelid muscles have been found. The aim of this paper was to study the presence and distribution of troponin in different muscle cell types from the earthworm Eisenia foetida (the muscular body wall, and the inner and outer muscular layer of the pseudoheart). These results were compared with those obtained in the transversely striated muscle of Drosophila melanogaster and in skeletal and smooth muscles of the mouse. METHODS: Immunocytochemical electron microscopic study and Western blot analysis using anti-TnT antibodies were employed in this study. RESULTS: Troponin immunoreaction was detected in the mouse skeletal muscle, the fly flight muscle, and earthworm obliquely striated muscles (body wall musculature and inner muscular layer of the pseudoheart). Immunolabeling for TnT in all these muscle cells appeared in moderate amounts at any point along the sarcomere length, except for the central zone of the A band (H band). This suggests that troponin molecules were located along the thin filaments. The density of immunogold particles was similar in the three muscles, and thus the amount of troponin in each muscle type was proportional to the number and length of actin filaments in each. Troponin was found in neither the mouse smooth muscle nor the outer muscular layer of the earthworm pseudoheart. The latter muscle showed an ultrastructural pattern that was intermediate between obliquely striated and smooth muscle. The estimated molecular weight for TnT in the earthworm was 55 kDa; this is higher than the weight of this protein in the mouse skeletal muscle (40 kDa) but similar to that of the D. melanogaster muscle (52 kDa). CONCLUSIONS: Troponin is present in both types of striated muscle (transversely striated and obliquely striated) of the earthworm with a distribution that is very similar to that observed in the mammalian striated muscle. As in vertebrates, troponin is absent in the smooth muscle of the earthworm. Discrepancies in the classification of some invertebrate muscles are common in the literature, and the use of distinctive markers, such as troponin, may improve our understanding of the nature and properties of many invertebrate muscles showing an ultrastructural pattern that does not resemble any of the classic muscle types.

Animals↗

A dynamic study of earthworm feeding ecology -using stable isotopes.

Changes in the specific diet of earthworms with time in relation to landuse changes and two different climates were studied by analysing (13)C and (15)N natural abundance in soils and animals. Soil samples from three depths (0-10, 10-20 and 20-30 cm) and earthworms were collected from two sites: Santiago (Northwest Spain) and North Wyke (Southwest England) both consisting of replicated long-term grasslands and recently converted to maize plots. Earthworms were hand-sorted in the field at the peak of the maize growth and after harvesting at both sites. In the Spanish plots, nine and eight earthworm species, all belonging to the three ecological categories (epigeic, anecic and endogeic), were found under maize and permanent pasture, whereas at the English site five and seven different species were, respectively, identified. At both sites (13)C isotopic values of the earthworm tissues reflected changes in diet from C(3) to C(4) with epigeic and epi/anecic worms in the maize plots showing one delta unit difference in relation to the ones found in the grassland plots. Anecic worms seemed to be less responsive to landuse changes. The higher (13)C values of the Spanish soils were also reflected in the earthworm tissues when compared with the English samples. (15)N values showed no clear relationship with the cropping treatments but were clearly related to the ecological grouping, with endogeic worms reaching the highest values whereas for the epigeic and epi/anecic species the lowest values were obtained. This finding was also previously recorded by other authors1 and suggests that, in the future, stable isotope techniques could also be a useful tool in taxonomic studies. Copyright 1999 John Wiley & Sons, Ltd.

Journal Article↗

Relevance and applicability of a simple earthworm biomarker of copper exposure. II. Validation and applicability under field conditions in a mesocosm experiment with Lumbricus rubellus.

Earthworms Lumbricus rubellus were exposed to a range of increasing soil copper concentrations in a mesocosm experiment for 17, 40, 70, and 110 days, respectively. Neutral-red retention times were measured along with earthworm tissue copper residues and earthworm growth and survival. The neutral-red retention assay demonstrated a clear dose-response threshold preceding a copper exposure value where increased regulatory activity had resulted in the copper bioconcentration factor (BCF) decreasing from 1 to 0.5. Effects on earthworm growth and survival occurred only at soil copper concentrations where the earthworm BCFs were lowered to ca. 0.5. Thus, the position of the neutral-red threshold could potentially make it possible to differentiate between copper exposure and actual toxicity. In terms of field applicability it was demonstrated that natural and seasonal variations in climatic parameters had little or no effect on the neutral-red response. Neutral-red retention assay has a potential role in environmental risk assessment and routine monitoring, as it is likely to provide a sufficiently accurate warning of impending ecological damage at a level below that found to cause actual physiological damage to the earthworms.

Analysis of Variance↗

Properties of tropomyosin from the dual-regulated obliquely striated body wall muscle of the earthworm (Lumbricus terrestris L.).

The obliquely striated body wall muscle of the earthworm Lumbricus terrestris L. possesses a dual actin-linked and myosin-linked regulatory system. Tropomyosin from this muscle has now been purified and its functional properties compared to tropomyosin from vertebrate skeletal muscle. Earthworm tropomyosin has a molecular weight of about 70 000 and is composed of two polypeptide chains of molecular weight of 34 000 and 37 000. Structural and functional similarities to skeletal muscle tropomyosin were demonstrated with respect to the formation and periodicity of paracrystals and nets and the potentiation of skeletal muscle acto-SF1 ATPase activity at low ATP concentration. Likewise, earthworm tropomyosin inhibited skeletal muscle acto-HMM ATPase activity at normal ATP concentrations but to a much greater extent than skeletal muscle tropomyosin; this inhibition was removed by skeletal muscle troponin, in the presence of Ca2+. In a system containing earthworm myosin and skeletal muscle actin, earthworm tropomyosin had no detectable influence on the actin-activated ATPase activity. It is concluded that earthworm tropomyosin plays an active role in the actin-linked troponin-dependent regulatory system and has no measurable effect on the regulation via myosin.

Actins↗

Geographic variation of freeze-tolerance in the earthworm Dendrobaena octaedra.

Freeze-tolerance and some of the underlying biochemical defence mechanisms in the earthworm Dendrobaena octaedra was investigated. Survival after slow cooling to -2 degrees C, -4 degrees C, or -6 degrees C was analysed in D. octaedra from three geographic regions representing large differences in winter temperature (Denmark, Finland and Greenland). A large variation in freeze-tolerance between the three populations of D. octaedra was found. Earthworms from the northern populations (Finland and Greenland) tolerated lower temperatures (-6 degrees C) than earthworms from the Danish population (poor survival at -4 degrees C and -2 degrees C). In the Finnish population, freezing led to the production of high concentrations of glucose, which reached values much higher than controls (94 mg g(-1) vs. 2 mg g(-1) dry weight). Other potential cryoprotectants were not elevated after freezing. The Danish and Greenlandic populations had substantially lower mean glucose levels after freezing than the Finnish population (about 15 mg g(-1)). Danish earthworms rapidly frozen did not accumulate glucose, and did not survive freezing at -2 degrees C. Danish earthworms exposed to osmotic stress in Ringer's solutions, containing different concentrations of glycerol, showed significantly elevated glucose levels, but did not survive rapid freezing. It was determined if freezing had an influence on the reproduction of the earthworms. After warming to summer temperatures (15 degrees C), survivors of freezing produced viable cocoons. In a field experiment it was tested if natural acclimatization during autumn and winter months had an effect on freeze-tolerance in the Danish population. There was a significant increase of post-freeze survival during this period. The results of the freezing experiments are discussed in relation to the general ecology of D. octaedra.

Acclimatization↗

Preliminary characterization of response-eliciting components of earthworm extract.

Fifteen garter snakes (Thamnophis sirtalis) reliably spent more time and tongue flicked more frequently at a dish containing earthworm extract than a dish containing distilled water when these were presented simultaneously for two minute intervals. The discriminability of the earthworm extract was directly related to its concentration. Garter snakes with their vomeronasal ducts sutured closed did not respond differentially to earthworm extract and water under these test conditions. Thus their ability to discriminate earthworm extract from water in this bioassay was dependent upon a functional vomeronasal system. Earthworm extract retains its biological activity after boiling at 100 degrees C for 15 minutes and after lyophilization. Its effectiveness is not altered by changes in pH. Snakes continued to respond differentially to extracts with pH 2, 5-6 or 11. Chloroform extractions of the acid, neutral and alkaline earthworm extract yielded activity primarily in the water layer. The small amount of activity in the chloroform layer was removed by use of a drying agent. Bradford dye-binding tests indicated the presence of protein in the active fractions.

Animals↗

Development of a standardized reproduction toxicity test with the earthworm species Eisenia fetida andrei using copper, pentachlorophenol and 2,4-dichloroaniline.

This article describes a standardized test method for determining the effect of chemical substances on the reproduction of the earthworm Eisenia fetida andrei. It is based on the existing guidelines for acute toxicity testing with earthworms, and for reasons of standardization the same artificial soil substrate and earthworm species were chosen as prescribed by these guidelines. After being preconditioned for one week in untreated soil, earthworms are exposed to the chemical substances for 3 weeks. The number of cocoons produced is determined, and cocoons are incubated in untreated artificial soil for 5 weeks to assess hatchability. Results are presented from toxicity experiments with pentachlorophenol, copper, and 2,4-dichloroaniline. For these compounds no-effect levels (NEL) for cocoon production were 32, 60-120, and 56 mg.kg-1 dry soil, respectively. Hatching of cocoons was influenced by pentachlorophenol (NEL, 10 mg.kg-1), but not by copper and dichloroaniline. Following exposure, earthworms were incubated in clean soil again to study the possibility of recovery of cocoon production. For copper and dichloroaniline earthworms did recover cocoon production to a level as high as the control level or even higher; in case of pentachlorophenol, cocoon production was still reduced after 3 weeks in clean soil.

Aniline Compounds↗

Earthworms as biological monitors of cadmium, copper, lead and zinc in metalliferous soils.

Earthworms (Lumbricus rebellus and Dendrodrilus rubidus) were sampled from one uncontaminated and fifteen metal-contaminated sites. Significant positive correlations were found between the earthworm and 'total' (conc. nitric acid-extractable) soil Cd, Cu, Pb and Zn concentrations (data log1) transformed). The relationships were linear, and the accumulation patterns for both species were similar when a single metal was considered, even though there were species difference in mean metal concentrations. Generally, the earthworm Cd concentration exceeded that of the soil; by contrast, the worm Pb concentration was lower than the soil Pb concentration in all but one (acidic, low soil Ca) site. Our observations suggest that Cu and Zn accumulation may be physiologically regulated by both species. Total-soil Cd explained 82-86% of the variability (V2) in earthworm Cd concentration; 52-58% of worm Pb and worm Zn concentrations were explained by the total-soil concentrations of the respective metals. Total-soil Cu explained only 11-32% of the worm Cu concentration. The effect of soil pH, total Ca concentration, cation-exchange capacity (CEC) and organic carbon on metal accumulation by L. rubellus and D. rubidus was investigated by multiple regression analysis. Soil pH (coupled with CEC) and soil Ca had a major influence on Pb accumulation (V2 of worm Pb increased to 77-83%), and there was some evidence that Cd accumulation may be suppressed in extremely organic soils. The edaphic factors investigated had no effect on Cu or Zn accumulation by earthworms. In the context of biomonitoring, it is proposed that earthworms have a potential in a dual role: (1) as 'quantitative' monitors of total-soil metal concentrations (as shown for Cd); and (2) as estimators of 'ecologically significant' soil metal, integrating the effects of edaphic factors (as shown for Pb).

Journal Article↗

Biomarkers responses of the earthworm Eisenia fetida to acetochlor exposure in OECD soil.

To examine the potential of a suite of biomarkers as early warning indicators of environmental pollution, sperm count, neutral red retention time (NRRT) and DNA damage were measured in earthworm Eisenia fetida exposed to increasing concentrations of acetochlor in OECD soil. The neutral red retention time of earthworms coelomocytes was sensitive to acetochlor pollution, and decreased significantly when the concentration was more than 10mgkg(-1) after 30 and 60 days of exposure (P<0.05). The reduced neutral red retention time correlated with the soil acetochlor residual. Sperm count decreased significantly at the concentrations of 40 and 80mgkg(-1) after 15 days of exposure (P<0.05). The DNA damage of earthworms coelomocytes increased significantly after 30 days of exposure at the highest concentration (80mgkg(-1); P<0.05). Earthworms were under physiological stress at field dose of acetochlor (10mgkg(-1)). Higher concentrations of acetochlor caused sperm count decrease and DNA damage of earthworms. Such a suite of biomarkers could serve as indicators of the health of the soil environment and to evaluate the toxicity of acetochlor on earthworms or as a means of monitoring soil acetochlor pollution.

Animals↗

Biomarker response and biomass change of earthworms exposed to chlorpyrifos in microcosms.

Background levels of chlorpyrifos and earthworm abundance were determined in an orchard and adjacent areas on a farm in the Western Cape, South Africa before these areas were again sprayed with this organophosphate. The background concentrations ranged from 0.2 microg/kg dm in the spray drift area adjacent to the orchard to 10.18 microg/kg dm on the slope in the run off area. In the target area the chlorpyrifos concentrations varied from a mean of 15.25 +/- 10.0 microg/kg directly after spraying to a mean of 7.0 +/- 0.9 microg/kg 6 months later and in the nontarget area they varied from a mean of 55.0 +/- 35 microg/kg to 12.0 +/- 5 microg/kg after 6 months. Chlorpyrifos was therefore still present in the field soils, but at lower concentrations, up to 6 months after the last spraying event. Earthworm abundance and population densities were very low. Only Aporrectodea caliginosa was found and the densities were much lower in the orchards (22 per m(2)) than in the nontarget areas (98.3 per m(2)). Microcosm studies were undertaken to relate biomarker responses to chlorpyrifos with biomass changes. Microcosms were filled with soil from the same areas and earthworms of the species A. caliginosa were introduced. The microcosms were treated with a series of concentrations of chlorpyrifos in the laboratory under controlled conditions. These concentrations were chosen to fall within the background ranges found in the soils. The biomass of the worms was determined regularly for a period of 5 weeks and worms in a state of estivation were noted. Earthworms were removed from the microcosms for biomarker tests: for cholinesterase (ChE) inhibition assays every week and for a neutral red retention determination 2 weeks after the exposures started. The most prominent biomass loss was noted in earthworms exposed to the highest pesticide concentration of 8.0 microg/kg. Estivation was higher among earthworms exposed to higher exposure concentrations. Inhibition of ChE increased with higher exposure concentrations and with time but there was no clear dose-related response. A clear dose-related response with exposure concentration was established for the neutral red retention assay. A correlation between ChE inhibition and biomass change existed directly after the second application of chlorpyrifos.

Animals↗

A review of studies performed to assess metal uptake by earthworms.

Earthworms perform a number of essential functions in soil; the impacts of metals on earthworms are often investigated. In this review we consider the range of earthworm species, types of soil and forms of metal for which metal uptake and accumulation have been studied, the design of these experiments and the quantitative relationships that have been derived to predict earthworm metal body burden. We conclude that there is a need for more studies on earthworm species other than Eisenia fetida in order to apply the large existing database on this earthworm to other, soil dwelling species. To aid comparisons between studies agreement is needed on standard protocols that define exposure and depuration periods and the parameters, such as soil solution composition, soil chemical and physical properties to be measured. It is recommended that more field or terrestrial model ecosystem studies using real contaminated soil rather than metal-amended artificial soils are performed.

Animals↗

Effects of perchlorate on earthworm (Eisenia fetida) survival and reproductive success.

The effects of perchlorate on earthworm, Eisenia fetida, survival and reproductive success were evaluated using three types of tests: dermal contact (filter paper), sand, and artificial soil. All studies utilized a range of perchlorate concentrations in order to simulate levels that are likely to occur in the environment under different scenarios (typical soil levels vs. spill levels). The OECD filter paper contact test involved exposing earthworms to the test compound on moist filter paper for 14 days to evaluate earthworm survival under a worst-case dermal exposure scenario. A similar test involved exposing the earthworms to perchlorate-contaminated sand, where earthworm survival decreased as concentration of perchlorate increased, with no worms surviving 14 days at the highest treatment concentrations (>2000 microg/g). However, the perchlorate concentrations that affected the survival of E. fetida are likely to occur only under extreme conditions (e.g. spills). The effect of perchlorate on the reproductive success (cocoon production) of E. fetida over a 4-week test period in artificial soil and a 3-week test period in sand was also examined. Production of cocoons was observed in soil containing up to 100 microg/g perchlorate, with no production in the uppermost treatment groups (1000 microg/g). Cocoon production was highest in the control group, although overall cocoon production appeared to be low. In contrast to the acute toxicity tests, perchlorate did affect earthworm reproduction at environmentally relevant soil concentrations. In addition, preliminary data suggest that cocoons produced under perchlorate contamination did not hatch as well as cocoons produced in control soil despite incubation of both sets of cocoons in clean soil or sand.

Animals↗