Lumbricus
- Lumbricus spp.
- Lumbricus castaneus
- Lumbricus friendi
- Lumbricus rubellus
- Lumbricus terrestris
[ref. ID; 6769]
Test system
Toxicants and worms
Soil were sampled around the chemical factory Kali Chemie AG in Bad Wimpfen, northern Baden-Wurttemberg. It was found in the 19th century as a salt works, but the upcoming aluminium industry prompted the management to change to kryolith production in 1921. From 1960 the factory diversified its F-chemistry prodution, including hydrofluoric acid, refrigerants, and spray propellants. In 1983, the factory was emitting about 2.8 kg/day, as HF. F emission and pollution of the surrounding landscape have a 67-years-old history.
The earthworms were extracted from the soil.
Measurements
F content of animals with gut and the F in chloragogen tissue.
[ref. ID; 5994]
Test system
Effect of urbanization on earthworm community
Study sites
The study was carried out in Brussels, Belgium. A transect was demarcated along heavily trafficked streets running from the city centre to the suburbs, in which a decreasing gradient of urbanization was predicted. Within the transect, six public parks were chosen for the study.
Sampling method
Earthworms were extracted by an electrical octet method (Thielemann, 1986).
Measurements/observations
Average density and biomass of earthworms at individual sites, Pb, Cd, Cu, Zn, Ca, and Mg concentrations of worms.
[ref. ID; 5996]
Test system
Pb uptake by earthworms
Sample
Earthworms were collected from 2 sites.
- Site 1. Near an old lead smelter in Tikkurila (The soil there is heavily contaminated with Pb from the smelter, which was in operation for nearly 60 years, until it was closed in the early 1980s and finally demolished in 1991).
- Site 2. Pornainen (less Pb contaminated situated ‘control’ soil) about 25 km to the north-east of Tikkurila.
Sampling method
The worms were extracted by hand-sorting on a plastic sheet.
Test design
- Experiment 1. Identification, counting and weighting of collecting earthworms.
- Experiment 2. Rearing experiments. Blocks of highly Pb contaminated soil were thawed and vertically cut into smaller units without destroying their structure, and the units were fitted into cylindrical plastic pots (diameter 8.5 cm, height 15 cm) to be used for rearing. The pots were fitted with caps of fine nylon net. During the rearing period the pots were kept inside a dark, ventilated chamber at temperature of +10 degrees C.
Measurements/observations
The number of species, the number of individuals and biomass, Pb concentration in earthworms and soil.
[ref. ID; 5990]
Test design
The heavy metal concentrations of copper, lead, cadmium and zinc in earthworms and soil from the border of a road.
Samples
Samples were collected in a meadow located at the border of a hairpin bend next to Osebe (U.T.M. 29TNH34) in the arterial road 550 and which bore traffic of about 10,000-15,000 vehicles a day.
Sampling method
Earthworms were collected from the soil by digging and handsorting.
Measurements
Concentrations of Cu, Pb, Cd, and Zn in soil and earthworm tissue.
Litter dwelling epigeic species. (ref. ID; 6138)
[ref. ID; 505]
Test system
Acute lethality tests (OECD 1984/EEC 1985)
Strains
Sexually mature.
Toxicants and Reference standard chemical
Chlorpyrifos and chloracetamide (ClCH2CONH2)
Test design
- Lethality test (14 days-LC50): Ten worms were placed in glass containers with 500 g dry mass of a soil-like substrate composed of a mixture of sphagnum peat, kaolinite clay, industrial quartz sand in a dry-weight ratio of 1:2:7. The pH (1N KCl) was adjusted to 6.0+/-0.5 with calcium carbonate. The water content was kept at 55% on a dry mass basis. Temperature 15 degrees C. Forty worms were tested at each test concentration.
- Reproduction test (14 days-EC50): Similar above experimental conditions, however, some modification following; a) the replacement of the artificial soil with a natural sandy soil (Kooyenburg) containing 3.7% organic matter, 1.4% clay, and pH (1N KCl) 4.8, and b) the supply of coarsely groud air-dried leaves of alder, Alnus glutinosa, for food.
Measurements/observations
Body weight and number.
Evaluations
14 days-LC50 according to the trimmed Spearman-Karber method (Hamilton et al. 1977). 14 days-EC50 using the statistical software package GENSTAT 5. NOEC applying analysis of variance.
[ref. ID; 1433]
Test system
Worm sampling site
- (i) The roadside verges of the A660 road in Leeds, West Yorkshire.
- (ii) The roadside verges of the A1 road in West Yorkshire.
- (iii) The verges of a service road through the University of Leeds farm (Spen Lane), at a site 300 m from the site by the A1 road.
- (iv) A city recreational area of deciduous woodland in Leeds (the Ridge), situated near a number of industrial sites.
- (v) A control site at Glen Sheilach, Oban, Scotland.
Measurements
Amounts of cadmium, lead, copper and iron in samples.
[ref. ID; 1913]
Test system
Accumulation test under laboratory and field conditions
Strains
From a local vermiculturist.
Toxicants
Copper.
Test design
Soils from an artificially contaminated (by applying CuSO4 at quantities of 0, 250, 500, and 750 kg Cu/ha) agricultural field near Wageningne, The Netherland. The soil is a slightly loamy fine sand and is low in organic matter content (loss-on-ignition: 3.5%). The soil was classified as a Fimic A Horizon (FAO-UNESCO, 1988) or a Plaggen Epipedon according to the USDA Soil Taxonomy.
Measurements/observations
Cu concentration in worm tissue.
[ref. ID; 1914]
Test system
The structure of model
Toxicants
Copper.
[ref. ID; 3580]
Test system
Strains
L. rubellus and D. rubidus from Carrock Fell, from Devon Great Consols and from uncontaminated site were collected by hand sorting.
Toxicants
Sodium arsenate. TWo arsenic- and heavy metal-contaminated mine-soil sites, at Carrock Fell, Cumbria and Devon Great Consols Mine, Devon. Uncontaminated site (mixed deciduous woodland site (SD 4888574) on the Lancaster University campus).
Test design
- Earthworm burrowing rates: The methods of Robinson et al. (1991). The soil was placed into Petri dishes (210 mm i.d.) with one half containing the uncontaminated soil and the other containing the same soil treated with 0, 12, 25, 123, 247, 494 or 1235 mg As kg-1 as sodium arsenate, with a divider separating the two soils. Earthworms (n=3) introduced onto the soil in each side of the dish. Six replicates. The time taken by earthworms to burrow entirely into the soil (15 mm depth) was recorded at 1, 3, 5, 10, 15, 20, 25 and 30 min.
- Soil discrimination: Soil was placed in Petri dishes with no divider separating the two soils. Earthworm (n=3) were introduced to each side of the dish. Replicates (n=3) for each species at each site and each concentration of sodium arsenate were stored in the dark at 9 degrees C.
- Arsenate toxicity: The uncontaminated soil was treated with sodium arsenate heptahydrate (494 mg As kg-1). Moistened soil (69 g) was weighed into the each of a series of 20x25 cm polythene bags. One earthworm was weighed and introduced in to each bag.
- Lethal concentration of As (LC50): The uncontaminated soil and Devon Great Consols were weighed and 55 g placed into a series of 20x25 cm polythene bags. Solutions of sodium arsenate were added to soil, setting the moisture content of the soil to 45%. Six concentrations of sodium arsenate were used for each site: Devon Grate Consols 247, 494, 988, 1235, 1976 and 2350 mg As kg-1 and 12, 25, 37, 74, 123 and 180 mg As kg-1, for the uncontaminated site. One earthworm was weighed and introduced into each bag. The bags were sealed and kept in the dark at 9 degrees C. Six replicates. Exposure period 14 days.
Measurements/observations
- Soil discrimination: Earthworms that had burrowed into each siil were recorded after 3 h.
- Arsenate toxicity: Specimens were examined at weekly intervals and assigned a conditions index (C.I.) score: 0 = dead, 1 = moribund (flaccid and/or unresponsible to tactile stimulation) and 2 = responsive (active and responsive to tactile stimulation).
- Lethal concentration of As (LC50): Conditions of alive or dead.
Evaluations
- Soil discrimination: A three factor ANOVA was carried out with species, concentration and site as factors, and a GLM model was constructed using Minitab.
- Lethal concentration of As (LC50): LC50.
[ref. ID; 4481]
Test system
Accumulation and elimination
Toxicants
Sodium arsenate heptahydrate (Na2HAsO4/7H2O).
Test design
- Contaminated site: From arsenic-contaminated sites at Devon Great Consols, an abandoned copper and arsenic mine near Tavistock, Devon, UK.
- Uncontaminated site: From a mixed deciduous woodland soil at Lancaster University campus (Lancaster, UK).
Measurements/observations
As concentration in worm tissue.
[ref. ID; 4581]
Test system
Uptake routes, ingestion (oral) or skin (dermal)?
Strains
From a non-polluted forest soil in Lepelstraat, The Netherlands.
Test design/concentrations
Orally sealed by medical histoacryl glue (Braun aesculap, Germany) and unsealed organisms.
- Water exposure experiment: The inert sand (quartz sand 200 um to <2 mm) matrix flushed with natural contaminated water continuously.
- Soil exposure experiment: Two metal-contaminated field soil.
Measurements/observation
Surviving number, weight and metal (Cd, Cu, Pb and Zn) concentration of body.
[ref. ID; 4639]
Test system
Accumulation
Strains
From studied site. Mature individual (clitellates).
Test design
Studied site: The site studied is located in Nord-Pas-de-Calais, northern France. In this area, there are two of the most important Pb an Zn smelters in Europe. The soils have been contaminated by wastes and dust emissions from the metallurgical industry since the middle of the 19th century. 6 sampling site.
Measurements
Contents of the metals (Pb, Zn, Cu, Cd) in earthworm tissues.
Evaluations
Biota-to-Soil Accumulation factor (BSAF).
[ref. ID; 5978]
Test system
Effect of pH on metal uptake and toxicity for earthworm
Strains
From a field-collected source and kept for one month in an uncontaminated culture medium. Mature adults weighing 500 to 1,500 mg.
Temperature/light condition
15+/-1.5 degrees C, 16:8-hr light:dark photoperiod.
Test design
Soils were collected from three sites under the deposition plume of a primary Cd, Pb, and Zn smelter (located at Avonmouth, southwestern England), located at different distances (site 1, 8.2 km; site 2, 3.2 km; site 3, 1.5 km) along a transect from the smelter.
1.5 L of test soils was placed into the experimental containers (polypropylene ice-containers, 180x180x93 mm), three pH treatment (Unamended, pH lowered by one unit (pH -1), pH increased by one unit (pH +1)) x 4 replicates, 8 worms/each replicate. At the start of the experiment, 5 g (dry wt) of suitable food (dried horse manure rewetted to 80% moisture content) was added to each container.
Measurements/observations
Mortality, number of cocoon, metal-binding protein metallothionein-2 (MT-2).
[ref. ID; 5984]
Test system
The influence of biotic factors of metal accumulation (31-days)
Strains
Worms collected from a single uncontaminated and ten contaminated sites in Mid-Wales, South East Wales and Shropshire.
Temperature/light condition
11 degrees C, dark.
Test design
Plastic container (21 cm x 12 cm x 18 cm).
- Run 1: Contaminated soil + worm collecting from same site.
- Run 2: Contaminated soil + worm collecting from uncontaminated site.
Measurements/observations
Mortality. Ca, Cd, Cu, Pb, and Zn concentration in earthworm tissues.
[ref. ID; 5987]
Test system
Effect of pH and calcium on lead and cadmium uptake in water
Strains
Subadult, 500 mg fresh weight. Prior to exposure they were starved for 2 days at 15 degrees C on moist filter paper to remove the intestinal contents.
Toxicants/concentrations
Pb(NO3)2 (concentration 0, 2.4, 9.7, and 24.1 uM Pb), Cd(NO3)2 (concentration 0, 8.9, 35.6, 89.0 uM Cd).
Test design
The earthworms were exposed singly in glass containers with 100 ml of aerated reconstituted water (100 mg NaHCO3, 20 mg KHCO3, 200 mg CaCl2/2H2O, 180 mg MgSO4 per liter demineralized water: pH 8.2) resembling natural groundwater for 24 hr and 15 degrees C.
- Run 1: Three concentrations of Calcium chloride (CaCl2: 0, 0.136, 1.36 mM Ca).
- Run 2: In the lead experiment, pH 4.0, 5.2, 5.7, 7.3 and 8.1. In the cadmium experiment, pH 4.0, 5.1, 5.8, 7.0 and 8.1.
Measurements/observations
Lead and cadmium of earthworm body were measured with using an atomic absorption spectrophotometer.
[ref. ID; 5989]
Test system
Influence of soil pH and organic matter for uptake of cadmium, zinc, lead, and copper by earthworms
Soil and worms
Sampling was done largely in the Dutch Kempen region located south-east of Eindhoven and north to a complex of several large zinc smelting works. The average weight of the worms with emptied gut contents was 86+/-28 mg (dry wt).
Measurements/observations
Samples of soil and worms were digested with nitric and sulfuric acids and analyzed for contents of Cd, Zn, Pb, and Cu by atomic absorption spectrophotometry.
[ref. ID; 5991]
Test system
Biological monitor
Collection sites
Earthworms were collected in October 1983 from heavily contaminated soils in the vicinity of disused non-ferrous metalliferous mines in Avon, Shropshire, Derbyshire and throughout Wales. Dinas Powis, a relatively uncontaminated site, was chosen as a control.
Measurements/observations
- Soil: pH, organic carbon, CEC, conc. nitric acid extractable Cd, Cu, Pb and Zn concentration.
- Worms: Dry-weight, tissue metal (Cd, Cu, Pb and Zn) concentration.
[ref. ID; 5992]
Test system
Heavy metal concentrations in the tissues, ingesta and faeces
Collection sites
Earthworms were collected by formalin extraction (20 L, 0.55%) from five separate 1 m2 sites situated on a longitudinal transect across the Cefn Parc Pb and Zn-mine, South Wales (O.S. grid ref. = ST 048822).
Test design
Earthworms were starved on moistened filter paper for 4 days by which time the alimentary canal was clear of ingested soil material. Animals were wet oxidized with concentrated (16N) "Analar" nitric acid, and analysed for Cd, Cu, Pb, and Zn by flame atomic absorption spectrophotometry.
Measurements/observations
Dry weight and tissue metal concentration of worms, metal concentrations of ingesta (crop contents) and egesta (faeces).
[ref. ID; 5993]
Test system
Cd uptake by earthworm
Strains
Field collected earthworm.
Toxicants
Cd(NO3)2.
Test design
An uncontaminated sandy soil from the Wildekamp field near Wageningen, The Netherlands, was spiked with cadmium.
Measurements
Number, weight, and Cd concentration of worms, cocoon number.
[ref. ID; 5994]
Test system
Effect of urbanization on earthworm community
Study sites
The study was carried out in Brussels, Belgium. A transect was demarcated along heavily trafficked streets running from the city centre to the suburbs, in which a decreasing gradient of urbanization was predicted. Within the transect, six public parks were chosen for the study.
Sampling method
Earthworms were extracted by an electrical octet method (Thielemann, 1986).
Measurements/observations
Average density and biomass of earthworms at individual sites, Pb, Cd, Cu, Zn, Ca, and Mg concentrations of worms.
[ref. ID; 5996]
Test system
Pb uptake by earthworms
Sample
Earthworms were collected from 2 sites.
- Site 1. Near an old lead smelter in Tikkurila (The soil there is heavily contaminated with Pb from the smelter, which was in operation for nearly 60 years, until it was closed in the early 1980s and finally demolished in 1991).
- Site 2. Pornainen (less Pb contaminated situated ‘control’ soil) about 25 km to the north-east of Tikkurila.
Sampling method
The worms were extracted by hand-sorting on a plastic sheet.
Test design
- Experiment 1. Identification, counting and weighting of collecting earthworms.
- Experiment 2. Rearing experiments. Blocks of highly Pb contaminated soil were thawed and vertically cut into smaller units without destroying their structure, and the units were fitted into cylindrical plastic pots (diameter 8.5 cm, height 15 cm) to be used for rearing. The pots were fitted with caps of fine nylon net. During the rearing period the pots were kept inside a dark, ventilated chamber at temperature of +10 degrees C.
Measurements/observations
The number of species, the number of individuals and biomass, Pb concentration in earthworms and soil.
[ref. ID; 5998]
Test system
The effect of exogenous (soil) calcium on lead accumulation by earthworms
Collection sites
Earthworms were collected from the shallow soil overlying the stony spoil heaps of 15 disused lead mines throughout Wales, Shropshire and Avon. Samples were also collected from an uncontaminated site (Dinas Powis) in South Wales.
Test design
Soil-feeding experiment: 3 soils were placed in a clean plastic box (23 cm x 23 cm x 12 cm) to a depth of 10 cm, respectively.
- 1. Cwmystwyth soil (a large volume of acidic, calcium-deficient and lead contaminated).
- 2. Calcium-supplemented (calcium concentration of approximately 6000 ug g-1 wet weight) Cwmystwyth soil.
- 3. Uncontaminated Dinas Powis soil.
15-20 worms were collected from Dinas Powis, were placed in the experimental boxes. The animals were maintained for 35 days in the dark at 4 degrees C, with soil moisture maintained at approximately 30%.
Filter paper feeding experiment:
Whatman No.1 filter paper was finely shredded in a liquidiser and 25 g were placed in six individual 500 ml glass beakers, the floors of which were covered by Whatman No.1 filter paper discs. To each baker was added 30 ml of a given test solution (Ca, Pb, and Ca + Pb). pH 5.4. 8-10 animals were used in each experimental group. The experimental was run in the dark at 15-17 degrees C for 14 days, and the filter paper and solution were changed daily.
Measurements/observations
Tissue lead concentration.
[ref. ID; 6065]
Test system
Bioaccumulation and biotransformation
Sampling sites
The field samples were collected from the area of the sawmill "Sikoniemi" (near the town of Kuopio in Central Finland). The sawmill was abandoned 28 yr ago, and at present there is a dense birch forest in the area.
Toxicants
2,3,4,6-Tetrachlorophenol.
Test design
Laboratory experiments: Soil was taken from a hayfield which had not been ploughed for several years (organic matter content 20% and pH 6.2) + 2,3,4,6-Tetrachlorophenol + food (clean birch leaf litter) + 13 worms, 16+/-1 degrees C.
Measurements
2,3,4,6-Tetrachlorophenol, Pentachlorophenol and their metabolites in worms.
[ref. ID; 6067]
Test system
Uptake of bound residues of Bentazone
Test design
10 individuals were held for 14 days in two soil types (sandy loam and loamy sand), both containing bound residues of [14]C-labelled (phenyl-u-[14]C) bentazone. The soil water content was adjusted to 17% (weight).
Measurements
Concentrations of [14]C-substances in different tissues of earthworm.
[ref. ID; 6083]
Test system
Metal accumulation
Sampling sites
- No. 1: High concentration of zinc (sewage sludge deposited on waste ground at Borth, Dyfed).
- No. 2: High concentration of lead (soil situated near the Cwmystwyth lead mine in the Ystwyth Valley).
- No. 3: High concentration of copper (waste tailing of the Glasdir copper mine near Dolgellau, Gwynedd, North Wales).
Measurements
Tissue concentration of Pb, Cu, Cd, Zn, Mn, and Ca.
[ref. ID; 6086]
Test system
The effect of calcium concentration on cadmium accumulation of two earthworms
Strains
Lumbricus rubellus and Allolobophora caliginosa were collected at Keele, Staffs.
Toxicants/concentrations
Cadmium (5 ppm).
Test design
The earthworms were placed in glass jars containing filter paper (Whatman No.1, 24 cm) for 21 days at 15 degrees, in the dark. At the end of the experimental period the filer paper was removed and the earthworms left for a further 5 days in contact with a small volume of cadmium solution.
Measurements
Cadmium concentration in earthworm tissues.
[ref. ID; 6090]
Test system
The influence of temperature and soil pH for sublethal toxic effect of copper on Lumbricus rubellus (6 weeks)
Strains
Adult worms were collected from a continuous grassland site.
Toxicants
Copper chloride (14 (unamended soil) - 372 mg Cu kg-1).
- Temperature: 12, 15, and 18 degrees C.
- pH: 4.8 (4.7-4.9), 6.0 (5.9-6.1), and 7.1 (7.1-7.2).
Test design
Experimental units were formed consisting of nylon-meshed netbags with a 5-litre volume of soil (agricultural sandy soil (loamy sand): 2% clay, 20% silt and 5.7% organic matter, pH (KCl) 4.8, soil moisture content 15%, and calcareous sandy loam soil: 17% clay, 74% silt, 3.4% organic matter, 5.5% CaCO3, soil moisture content 35%). 5 worms per unit. 5-8 replicates.
Measurements
Mortality, body weight, cocoon production, litter breakdown activity.
[ref. ID; 6095]
Test system
Calcium-lead interaction
Strains
Specimens were collected from the spoil-heap soils of the abandoned lead mines at Cwmystwyth (acidic, pH 4.3) and Draethen (calcareous, pH 6.5) in Mid- and South Wales, respectively. Also, L. rubellus was collected from Dinas Powis, an uncontaminated site in South Wales.
Toxicants
Lead.
Test design
Filter paper feeding experiment: 8-10 animals were exposed to finely shredded Whatman No.1 filter paper (25 g) soaked in 30 ml of test solutions (deionised water, 100 ug/ml Ca, 400 ug/ml Ca, 80 ug/ml Pb, 80 ug/ml + 100 ug/ml Ca, 80 ug/ml + 400 ug/ml Ca) in the dark at 15-17 degrees for 14 days. The filter paper and solutions were changed daily. pH 5.4.
Measurements
Lead concentration in worm tissue.
[ref. ID; 6102]
Test system
Seasonal changes in the tissue-metal (Cd, Zn and Pb) concentration in earthworm in heavy metal polluted soil
Sampling sites
Mature (clitellate) worm were collected monthly from a 9-m2 sampling site at Cefn Parc mine, near Llantrisant, S. Wales (O.S. grid ref. ST 048822).
Toxicants
Cd, Pb, Zn.
Measurements
Dry weight and Cd, Pb, and Zn concentration in worm.
Evaluations
Monthly differences in tissue-metal concentrations were evaluated statistically by Duncan's multiple-range test. Differences in tissue-metal burdens between diapause and pre- or post-diapause animals were statistically evaluated by the Mann-Whitney U-test.
[ref. ID; 6136]
Test system
Earthworm responses to Cd and Cu under fluctuating climate (temperature and rainfall)
Strains
Both adult (fifteen individuals: 500-1500 mg weight) and juvenile (ten individuals: mean weight 30 mg range 6-99 mg) were added to the mesocosms. Adults were obtained from a commercial source (originally field collected).
Toxicants
CuCl2/2H2O and CdCl2/2.5H2O.
Test design
Mesocosm design: Each unit comprised a 40 cm section of 30 cm internal diameter medium density polyethylene pipe with wall thickness of 12 mm. The bottom of each pipe was sealed using nylon and medium density polyethylene (mesh size 4 mm) and 64 g m-2 Phormisol (LBS Horticulture, Colne, UK). This was then screwed to the base of the mesocosm and secured with silicone sealant. The top of mesocosm secured 280 x 360 um mesh.
Test medium: A commercially available clay loam soil (Broughton Loam, Kettering, UK), with a pH of 7.1 and a 5% organic matter content. Fifteen kilograms dry weight of soil was used in each mesocosm with four replicates per treatment. For moistening and spiking, soils were placed in a watertight plastic bag within the mesocosm and aqueous solution of CuCl2/2H2O and CdCl2/2.5H2O added at volumes and concentrations sufficient to give a water content of 60% of soil water holding capacity and nominal metal concentrations of 0, 0.16, 0.63, 2.52, 7.56 umol Cu g-1 in the Cu test and 0, 0.111, 0.45, 1.78 and 5.34 umol Cd g-1 in the Cd test. The field plot was situated in the southeast, UK (Ordnance Survey Grids Reference TL 798292).
Total exposure time: 70 days.
Measurements/observation
- Metal residure analysis: Flame atomic absorption spectrophotometry of nitric acid digests according to the method on Spurgeon and Hopkin (1996).
- Lysosomal membrane stability: The nutral red retention time (NRR-T) assay.
- Metallothionein-2 (MT-2) transcript quantification: Real-time quantitative reverse transciptase polymerase chain reaction (Q-RT-PCR) using the fluorogenic 5' nuclease assay according to the procedure set out in Galay-Burgos et al. (2003).
Evaluations
- LC50 using the trimmed Spearman-Karbar method.
- Juvenile production EC50 (linear interpolation using the ICp program version 2.0).
[ref. ID; 6138]
Test system
OECD-style toxicity (28 days) test
Strains
From Ecology Earthworms, Hubbards Hall Farm, Bentley, Ipswich, Suffolk, UK. Mature adult (average weights 0.76+/-0.11 g), 280 individuals.
Toxicant/concentrations
Pb(NO3)2 1000, 3000, 4000, 5000, 7500 and 10 000 mg Pb kg-1.
Test design
OECD-style test: 1 L plastic containers (500g dry weight soil (Kettering loam, purchased from Barrycroft Stores Limited, Kettering, Cambridgeshire, UK), moisture content of the soil to 50% of the total WHC (water holding capacity), 15 degrees C, four replicates.
Measurements/observations
Mortality, weight of worms, total Pb in earthworm tissue.
Evaluations
- LC50 using Probit analysis on SPSS statistical software version 10.01.
- EC50 using USEPA (1984) Environmental Research Laboratory, Duluth MN 55804 USA, linear extrapolation method for sub-lethal toxicity.
[ref. ID; 6708]
Test system
Effect of pyrene for metabolomics
Strains
Adults weighing >500 mg.
Toxicity
Pyrene.
Test design
1 kg (dry weight) of soil mix was placed dry into 1 L Kilner jars and spiked with concentrations of 0, 10, 40, 160 and 640 mg kg-1 of pyrene in equal volumes of acetone. Soils were then vented for 72 hr to remove all the solvent, wetted to 60% of water holding capacity. Six worms added to each jar. For 42 days at 15+/-1.5 degrees C in a 16:8 hr light:dark. Food (air-dried horse manure).
Measurements/observations
1H nucelar magnetic resonance (NMR) analysis and gas chromatography mass spectrometry (GC-MS) analysis of worm tissue extraction.
[ref. ID; 6711]
Test system
Combined effects of zinc and earthworm density for risk assessment
Strains
Worms are collected at an unpolluted site (Nijkerkerveen, The Netherlands). Adult worm (average individual wet weight+/-SD: 1.58+/-0.11 g) density (0, 3, and 5 worms per containers).
Toxicants
ZnSO4/7H2O (0, 140, 300 and 620 mg Zn kg-1 dry wt). In order to check for effects of sulfate addition, CaSO4/H2O was added to a separate series of microcosms without worms at concentrations of 0, 172, 554 and 1721 mg CaSO4/2H2O kg-1 dry wt.
Test design
Soil: Soil was collected from the experimental organic farm 'Kooyenburg' at Rolde, The Netherlands. The soil is a sandy loam soil, slightly acidic and with a low SOM (soil organic matter) content. This soil has a long history of no-tillage farming without pesticide application. The soil contains low concentrations of heavy metals. The natural background concentration of zinc is 18 mg kg-1 dry wt.
Microcosms: The microcosms consisted of 1-l NALGENE cylindrical Perspex containers (diameter 115 mm, height 130 mm). They received 700 g air-dried soil, wetted with 114 ml demineralized water (soil moisture content of approx. 15% (w/w)). At the beginning of the experiment alder leaves (Alnus glutinosa (L.) Gaertn.) were added to the soil surface of each microcosms. 15 degrees C, relative air humidity of 80% and under permanent light conditions.
Measurements/observations
Respiration was measured every 2 hr in the headspace of each microcosm using conductometry.
After 52 days incubation, number and weight of worms measured. The weight of the remaining A. glutinosa litter was measured separately in each sieve fraction (mesh widths of 31.3, 16, 8, 4, 2, 1, 0.5 and 0.25 mm).
[ref. ID; 6713]
Test system
Flooding response
Strains
- For pot experiment: From grassland soil near Bergen op Zoom, in the southern part of the Netherlands.
- For another experiment: From the 'Afferdensche en Deetsche Waarden'(ADW), a floodplain system along the river Rhine, in the central part of the Netherlands.
Toxicants
Zinc, cadmium, copper.
Flooded soil: Soil was collected from the top 0-10 cm horizon in an 'Afferdensche en Deetsche Waarden'(ADW).
Test design
Pot experiment: 44 plastic flowerpots (18 cm diameter, height 18.5 cm) 100 mg kg-1 zinc (Zn(NO3)/3H2O), 5 mg kg-1 cadmium (Cd(NO3)/4H2O), 20 mg kg-1 copper (Cu(NO3)2/3H2O). On top of the soil, grass (Lolium perenne) was sown. At 12 degrees C with continuous light, during 42 days. In the flooded treatments, the water level was raised to 5 cm water above the soil surface.
- Run 1: Polluted combined with flooded soils + A. caliginosa 8 N and L. rubellus 3 N.
- Run 2: Non-polluted combined with flooded soils + A. caliginosa 5 N, L. rubellus 3 N, and A. chlorotica 3 N.
- Run 3: Polluted combined with non-flooded soils + A. caliginosa 8 N and L. rubellus 3 N.
- Run 4: Non-polluted combined with non-flooded soils + A. caliginosa 5 N, L. rubellus 3 N, and A. chlorotica 3 N.
Moisture preference experiment: 8 glass aquaria (31 cm x 19 cm x 20 cm) were dividing five equal compartments with four Perspex partitions (height 13 cm). The aquaria were filled to a depth 11 cm with field soil at different moisture contents (35%, 45% (field capacity), 55%, 65% (saturated), 65%+ (saturated and an extra water layer) w/w). At 12 degrees C with continuous light, during 9 days.
- Run 1: A. caliginosa 5 N (3 N adult + 2 N juveniles, FW 1.69 +/- 0.21 g per compertment).
- Run 2: L. rubellus 5 N (3 N adult + 2 N juveniles, FW 1.28 +/- 0.12 g per compertment).
- Run 3: A. chlorotica 3 N (2 N adult + 1 N juveniles, FW 0.51 +/- 0.08 g per compertment).
Health experiment: 40 small plastic buckets (diameter 12 cm, height 14 cm) were filled with 0.7 cm3 field soil. Two adult of each of the species A. caliginosa, L. rubellus and A. chlorotica added to each bucket (moisture content (35%, 45%, 55%, 65%, and 65%+). At 12 degrees C with 12h light, during 42 days.
Measurements/observations
Worm number and weight.
[ref. ID; 6741]
Test system
The mechanistic bioaccumulation model OMEGA (Optimal Modeling for Ecotoxicological Applications) predictions
Toxicants
Concentration of cadmium, copper, lead, zinc in the earthworm Lumbriucs rubellus, soil and porewater.
[ref. ID; 6743]
Test system
Effect of temperature and season
Strains
Worms obtained from a field collected source and then kept outdoors, under cover for one month in uncontaminated culture medium consisting of 33% loamy soil, 33% peat, 33% composed bark (LBS Horticultural, Colne, UK). Fully mature adults weighing 400-1600 mg used.
Soils
Soils were collected from three sites under the deposition plume of a primary Cd, Pb, and Zn smelter located at Avonmouth, South West England, and from three different seasons (spring, autumn, winter).
Test design
Plastic boxes with dimensions (180 mm x 180 mm x 93 mm) were filled with 1.5 L of the relevant soil. 8 worms added. Containers were covered to limit water loss and kept at the relevant temperature (10, 15 and 20 degrees C) in a 16 hr light 8 hr dark regime for 42 days. 5 g (dry weight) of food (dried horse manure re-wetted to 80% moisture content) was spread on the soil surface. Weekly, excess food was removed and 5 g of fresh food added.
Measurements/observations
Mortality, lysosomal membrane stability assay (NRR-T assay), metallothionein-2 protein, metal concentrations (As, Cd, Cu, Hg, Pb, Zn).
[ref. ID; 6744]
Test system
Effect of time and mode of depuration on tissue copper concentration
Strains
From Ecology Earthworms, Hubbards Hall Farm, Bentley, Ipswich, UK. Individuals with fully clitellate (mean fresh weights: 1158+/-1.56 mg).
Toxicant/concentrations
Cu(NO2)3/3H2O, 250 mg Cu kg-1.
Test design
Culture period 28 days.
- Soil: Kettering sandy loam soil is similar in its properties to the OECD standard soil. Moisture content 50%.
- Worm: Eighty earthworms were added to 12 kg of the treated Kettering loam. 15+/-1 degrees, constant darkness.
Depuration method: 1) Depuration time (0, 24, 36, 48, and 72 hr). After depuration the filter papers (Whatmann no.540) were dried at 40+/-1 degrees C for 24 hr and re-weighed. The earthworms were rinsed in ultrapure deionised water, dried on tissue paper and immersed for 20sec in boiling ultrapure water. 2) After depuration for 0, 24, 36, 48 or 72 hr, the earthworms was dissected following immersion in boiling water for 20sec. The earthworms were slit open to expose the alimentary canal, which was cut from above the crop to the bottom of the intestine. Each earthworm was pinned open onto a wax board and a small brush was used to carefully remove the soil particles present in the crop, gizzard and intestine.
Measurements/observations
The concentration of Cu and Ti in tissues of worm.
[ref. ID; 6745]
Test system
Metal bioaccumulation
Floodplain site
Three sites along the rivers Nieuwe Merwede and Waal. The sites are located on gradually sloping riverbands, and are subjected to periodic inundation.
Collection organisms
Collection was random within the 10x10 m plot. At least 5 individuals of each pedo-ecological group were taken. After taking to the laboratory, the earthworms were allowed to defaecate on wet filter paper for 48 hr. Wet weights were determined, followed by storage in freezer at -18 degrees C until analyses.
Measurements/observations
As, Cd, Cr, Cu, Pb, Zn, Mn, Fe, Ca concentration in earthworms.
[ref. ID; 6749]
Test system
The effect on heavy metal (Pb and Zn) fractionation of processing the soil through earthworm's digestive tracts, mobility, and oral-bioabailablity in earthworms casts.
Strains
Worms obtained from Biobrazda (Dragomer, Slovenia), were used fully clitellated adult specimens and subadult specimens with clear signs of developing tubercula pubertatis.
Toxicants
Soil was collected from the 0-30 cm surface layer of an abandoned vegetable garden in the vicinity of a former industrial site in the Mezica Valley in Slovenia. The Mezica Valley has been exposed to more than 300 years of active Pb mining and smelting.
Test design
Pot experiment: Clean plastic pots (height 9 cm, diameter 12.5 cm) were filled with 250 g of air-dried non-remediated and remediated soil, in three replicates. 80% of soil field water capacity. 10 earthworms (0.12-0.29 g fresh weight) introduced into each pot and kept in the dark at 20 degrees C for 7 weeks.
Lead bioavailability in warm casts was determined as oral bioavailability in simulated stomach (pH 2.50+/-0.05) and intestinal (pH 7.00+/-0.05) phases of human gastrointestinal tract, using Ruby's physiologically based extraction test.
Measurements/observations
Fractionation (assessed using sequential extractions) of Pb and Zn in worm casts.
[ref. ID; 6763]
Test system
Strains
Adult earthworms were provided by Edwin Berry, National Soil Tilth Laboratory, USDA Agricultural Research Service, Ames, Iowa. The earthworms species used were Lumbricus rubellus (weighing 0.80-1.21 g), Aporrectodea trapezoids (weighing 2.15-2.57 g), Lumbricus terrestris (weighing 5.08-6.23 g).
Toxicants
Plasmid: pJP4, Donor: Alcaligenes eutrophus JMP222N, Recipient: Pseudomonas fluorescens C5t
Test design
Soil: A Hubbard loamy sand (Udorthenic Haploboroll).
Microcosm: The microcosm consisted of a polyvinyl chloride tube (50 by 10 cm diameter) which was cut into eight 5-cm segments. The remaining 10-cm segment served as the headspace.
Experimental design
Soil microcosms containing spatially segregated, coinoculated donor and recipient strains, inoculated control (donor and recipient strain only) soil microcosms, and uninoculated control soil microcosms. Triplicate microcosms for each of the four treatment (i.e., no worms, L. rubellus, A. trapezoides, and L. terretris). 3 adult earthworms added to surface of the microcosm. (The final earthworms density was equivalent to approximately 100 earthworm per m2 of soil. 20 degrees C. Experimental period 2 weeks.
Measurements/observations
Vertical distribution of earthworms number. Donor, recipient, transconjugant bacteria number of casts and cocoon.
[ref. ID; 6786]
Test system
The applicability of CBRs as a practical tool in soil quality assessment of contaminated sites
Strains
From unpolluted grassland sites in the region of Wageningen. Clitellated adult stage.
Toxicants
CuSO4/5H2O.
Test design
Batches of five adults worms in 600 ml volume of soil (an arable sandy soil from Kooyenburg, and a silty clay loam from Oostelijk Flevopolder) were incubated in a climate chamber with a constant temperature of 15 degrees C and continuous light. Crushed leaves of alder (Alnus glutinosa) were added on top of the soil surface as a suitable food source. Experiment period was 4 weeks.
Measurements
Survival and cocoon number.
Evaluations
Treatment effects and dose-response data were analyzed using the Genstat 5, release 4.2, statistical package.
[ref. ID; 6787]
Test system
The effects of earthworm density on the life-history parameters
Strains
Adult were used to produce F1 cocoons.
Test design
Bioassay containers (64 one-liter containers were filled with 650 g of soil (sandy loam soil pH 5.1, organic matter 5.7%, clay content 2%). Crumbled alder leaves (Alnus glutinosa, about 80 g) were placed on top of the soil surface after being rewetted with distilled water. Containers were stored in controlled climate rooms at 15 degrees, 61% humidity, and in continuous light. Experiment period was 6 months. Food was added if the amount was reduced by about 50% such that food remained over-abundant during the test in all containers.
Juveniles (mean weight+/-SD, 16+/-2 mg) were allotted randomly to the 64 containers in earthworm densities ranging from two to nine earthworms per container, with eight replicates for each earthworm density.
Measurements
Number and weight of adult, subadult, and cocoon production.
[ref. ID; 6819]
Test system
Strains
From a reference (control) and two polluted (smelter and mine) sites. Adult worm.
- The reference strain was collected from the campus of the University of Reading, UK.
- The smelter strain was collected approximately 3 km from a smelting works situated at Avonmouth, UK (Hallen Wood). This site has been subject to aerial deposition of cadmium, copper, lead, and zinc since 1929.
- The mine strain was obtained from Shipham, UK. This site is contaminated with very high levels of cadmium, lead, and zinc as a result metal extraction and processing during the sixteenth and seventeenth centuries.
Toxicant/concentrations
ZnNO3/6H2O (0, 190, 350, 620, 1200, 2000, and 3600 ug Zn g-1).
Test design
Six worms were weighed and added to each test replicate (plastic boxes, 220 mm x 160 mm x 80 mm). Containers were covered and kept for 42 days at 15 +/- 2 degrees C under constant light. A suitable food (finely ground fresh horse manure, dried and rewetted to 75% water content) added. Artificial soil (OECD, 1984 and EEC, 1985) was not used, because previous experiments to measure metal toxicity for L. rubellus in OCED soil have indicated some problems, in particular high control mortality. Tests were conducted with a medium based on commercially available sady loam topsoil, and 20% by weight of finely ground Sphagnum peat was added.
Measurements/observations
Survival, weight change, cocoon production, and zinc concentration in worm.
[ref. ID; 6849]
Test system
Heavy metal accumulation
Toxicants
Sewage sludge (Milorganite, an anaerobically digested, heat-dried commercial sludge (N-P-K, 6-2-0)).
Test design
The study area consisted of eight 0.1 ha enclosures in the third year of secondary succession. The enclosures were ploughed, disced, and fertilized with 336 kg ha-1 commercial fertilizer (N-P-K, 12-12-12) in 1977. Winter wheat Triticum aestivum var. Ranger was planted in October 1977. The wheat matured in 1978 and was not harvested; the fields were allowed to go fallow in 1979.
Three replicates plots were treated with dried sludge, three with fertilizer, and two left as untreated controls.
Measurements/observations
Heavy metal (Cd, Cu, Pb, Zn) concentration in earthworm.
[ref. ID; 6858]
Test system
Life-cycle and biomarker responses to zinc in four earthworm species (Aporrectodea caliginosa, Eisenia fetida, and Lumbricus terrestris)
Strains
From an unpolluted pasture on the University of Reading Campus, Reading, United Kingdom, by digging and hand sorting. Adult worm with mean wet weight 732 mg.
Toxicants/concentrations
Zn(NO3)2/6H2O aqueous solutions: 0, 190, 350, 620, 1200, 2000, and 3600 ug/g.
Test design
Zinc exposures were conducted in a natural soil-based test system. The soil used was a mixture of a commercially available sandy loam soil (Rockalls, Wokingham, Berkshire, UK) and commercially available Sphagnum peat (Bullrush Ltd, County Tyrone, UK). One kilogram of the soil mix was added to each experimental a container (plastic boxes 220x160x80 mm), with four replicate containers used for each test concentration. Water-holding capacity 60%. 6 worms per container. During exposure period, the test containers were covered to limit water loss and kept in constant light at 15 degrees C for 42 days. Finely ground fresh horse manure (dried and rewetted to 75% water content) was added as source of food (4 g dry weight per weekly to each container) in all tests.
Measurements/observations
- Life-cycle parameter: Survival, weight change, and cocoon production rate.
- Biomaker: Neutral-red retention by coelomocytes lysosomes.
- Zinc conconcentration in worm tissues.
Evaluations
Significant differences in parameters were calculated using analysis of variance (ANOVA). When, differences were found, Tukey's multiple comparison test was used to determine differences between specific treatments.
- LC50 by the log-probit method using the MicroProbit 3.0 statistical software package.
- Sublethal sensitivity index (SSI) were determined from calculated effect concentration (EC10) values. SSI=LC50/EC10.
[ref. ID; 6860]
Test system
Inherited resistance
Strains
- Field-collected adults: Specimens of mature L. rubellus were collected by hand-sorting soil from a culture at the Centre for Ecology and Hydrology (Monks Eood, Huntingdon, UK) and an abandoned arsenic and copper mine (Devon Great Consols, Devon, UK), and an abandoned tungsten mine (Carrock Fell, Caldbeck, Cumbria, UK).
- F1 generation adults.
Toxicants
Sodium arsenate (Na2HAsO4/7H2O), copper chloride (CuCl2).
Test design
Soil from an uncontaminated, mixed deciduous woodland site on the Lancaster University campus was rewetted to a moisture content of 53% (dry wt equivalent) using a solution of sodium arsenate (Na2HAsO4/7H2O) to give a concentration of 2,000 mg As/kg or with a solution copper chloride (CuCl2) to give a concentration of 300 mg Cu/kg dry weight of soil. Mean soil pH 5.34+/-0.04. Moistened soil (69 g) was weighed into each of a series of 20x25-cm polythene bags.
One worm per each bag. The bags were kept for 28 days at 9 degrees C in 24 hr of darkness.
Measurements
Condition index scores, 0 = dead, 1 = moribund (flaccid unresponsive to tactile stimulation), or 2 = turgid (responsive to tactile stimulation). Number of cocooons produced in the field-collected adults and the F1 generation adults. Arsenic concentration in worm tissue.
Evaulations
Condition Index.
[ref. ID; 6892]
Test system
Effect of soil heterogeneity
Strains
From a vermiculturist.
Toxicants
CuSO4.
Test design
- Field experiment: The field (an agricultural field near Wageningen, The Netherlands) consisted of eight blocks of 16 plots each (6x11 m2). Cu concentration (for CuSO4) 0, 250, 500 and 750 Cu ha-1. pH 4.4, 4.7, 5.4 and 6.1. In the middle of the sampled area of each plot, about 500 earthworms, obtained from a vermiculturist, were introduced. The earthworms were sampled after 2, 4, and 10 wk by gathering 25 earthworms out of each plot.
- Laboratory experiment: About 120 kg above mentioned field soil was homogenized and split up into six portions of 20 kg each and put into plastic containers (40x30x20 cm3). Three water contents 25, 35, and 45%. These containers were put into a phytotron, adjusted to 12 hr daylight per day, room temperature at 15 degrees C, and relative air humidity of 85%. Exposure period 1, 3, 7, 14, 28 and 56 days.
Measurements/observations
Cu concentration in worm tissue.
[ref. ID; 6909]
Test system
Resistance to arsenic toxicity
Strains
- Run 1. 18 adult and large immature L. rubellus were collected by hand-sorting in mixed deciduous woodland.
- Run 2. L. rubellus were collected from an uncontaminated grassland site on the University of Lancaster campus (mean body weights: 1087+/-120 mg), and from Carrock Fell mine spoil (mean body weights 490+/-82 mg).
Toxicants
- Run 1. As contaminated soil. (Spoil at an abandoned tungsten mine at Carrock Fell, Cumbria, NW England, contains As, largely as arsenopyrite, in concentrations up to 53,000 mg kg-1).
- Run 2. Sodium arsenate heptahydrate 2000 mg kg-1 dry weight of soil of the hydrated salt.
Test design
Experimental period 28 days.
- Run 1. 500 g soil samples were weighed into 20x25 cm polythere bags and 200 mg of dried, ground mine spoil vegetation, dominated by grass and rush, was added as food. One earthworm was placed in each bag. The bags were sealed, stored in the dark at 9 degrees C and periodically opened for ventilation. Six L. rubellus were removed after 5, 10 and 15 days.
- Run 2. Uncontaminated soil + Sodium arsenate heptahydrate was weighed into each of a series of 20x25 cm polythene bags. One earthworm (from campus) and 10 earthworms (from Carrock) introduced into each bag. The bags were sealed and stored in the dark at 9 degrees. After 7 days, 200 mg dried, powdered nettle (Urtica dioica L.) leaves were added as a food supplement.
Measurements/observations
Tissue As concentration in L. rubellus and body weight.
Evaluations
Condition Index.
[ref. ID; 6912]
Test system
Tolerance to arsenic toxicity
Strains
Resistant strain were collected by hand-sorting from Devon Great Consols mine at Tavistock, Devon, UK. The strain show high tolerance to Cu- and As-toxicity and frequently have a striking yellow coloration.
Another strain were collected from the uncontaminated, mixed deciduous woodland on the Lancaster University campus.
Toxicants
The Devon Great Consols soil contained considerably higher concentrations of As 9845+/-40 mg kg-1 and Cu 1645+/-220 mg kg-1.
Test design
Moistened soil was weighed into each of a series of 20x25 cm polythene bags. One earthworm was weighed and introduced into each bag.
Measurements/observations
Pigmentation, As concentration in worm tissue and condition index.
[ref. ID; 6953]
Test system
Toxicity and bioaccumulation
Strains
Adults with a well-developed clitellum and average weight 1314-1816 mg.
Toxicants
3-Chlorophenol, 3,4-Dichlorophenol, 2,4,5-Trichlorophenol, 2,3,4,5-Tetrachlorophenol, Pentachlorophenol.
Test design
Soils: Soils was collected from the top 20 cm of agricultural fields. Holten soil (very humic sand) and Kooyenburg soil (moderately humic sand).
Glass jar filled with about 0.65 kg soil (dry wt) and 5 worm. The jars were placed in an incubator at 15 degrees C. Experimental period 7 and 14 days.
Measurements
Mortality.
Evaluations
LC50 values based on 14 days'mortality data were calculated according to a logit model.
[ref. ID; 6978]
Test system
2-wk LC50
Strains
Toxicants
3-chlorophenol (MCP), 3,4-dichlorophenol (DCP), 2,4,5-trichlorophenol (TCP), 2,3,4,5-tetrachlorophenol (TeCP), pentachlorophenol (PCP), 2,4-dichloroaniline (DCA), 1,2,3-trichlorobenzene (TCB).
Test design
Soil type: OECD artificial soil, two different sandy soil, and peaty soil.
Tests carried out at least in duplicate, with at least 5 concentrations and a control.
Evaluations
LC50 using trimmed Spearman-Karber method. Quantitative structure-activity relationships (QSARs).
[ref. ID; 7027]
Test system
Assessment of Equilibrium partitioning theory in in situ studies and water experiments
Strain
Toxicants
Benzo[a]pyrene, Fluoranthene, Phenanthrene, and Pyrene.
Test design
- Field study: Samples of soil and earthworms were collected from sites in the floodplains along the River Waal near Ochten and Gelderse Poort. Adult L. rubellus were extracted from upper 20 cm of soil by digging and hand sorting.
- Experimental study (water-only exposure): Adult specimens of L. rubellus with emptied guts were exposed to 200 ml of aqueous solution (NaHCO3 100 mg/L, KHCO3 20 mg/L, CaCl2 200 mg/L, and MgSO4 180 mg/L in demineralized water) in glass vessels. The experiments were conducted in total darkness to avoid photolytic degradation or phototoxic effects of the test componds. One worm was used per experimental vessel, 3 replicates.
Measurements/observations
- Field study; Total concentration of PAHs in worm.
- Experimental study: Concentration of Benzo[a]pyrene, Fluoranthene, Phenanthrene, and Pyrene in worm lipid.
Evaluations
BSAF, BAF, BCF.
[ref. ID; 7055]
Test system
The distribution of Cd, Cu, Pb, Zn, and Ca in the worm tissues
Samples
Mature (clitellate) L. rubellus were collected from an uncontaminated site (Dinas Powys, O.S. grid reference ST 146723) and four disused non-ferrous metalliferous mine sites (Llantrisant, ST 048822; Draethen, ST 217877; Cwmystwyth, SN 803748; Ecton, SK 098581). Worms and soils were collected from a 3-m diameter circle at each site.
Toxicants
Cd, Cu, Pb, Zn, and Ca.
Measurements
The worms were dissected following the tissue fractions.
- 1. The 'anterior' alimentary canal, i.e. the buccal cavity, oesophagus and associated calciferous glands, crop and gizzard
- 2. The 'posterior' alimentary canal, i.e. the intestine and encapsulating chloragogenous tissue
- 3. The remaining tissues, (including the body wall, reproductive tissues, and nephridia) designated the "rest".
Dry weight and metal content in each tissue fractions.
Evaluations
Tissue affinity for Cd, Cu, Pb, Zn, and Ca.
Anecic species. (ref ID; 6763)
Lumbricus terrestris is anecic and lives in deep vertical burrows. (ref. ID; 6858)
[ref. ID; 505]
Test system
Acute lethality tests (OECD 1984/EEC 1985)
Strains
Sexually mature.
Toxicants and Reference standard chemical
Chlorpyrifos and chloracetamide (ClCH2CONH2)
Test design
- Lethality test (14 days-LC50): Ten worms were placed in glass containers with 500 g dry mass of a soil-like substrate composed of a mixture of sphagnum peat, kaolinite clay, industrial quartz sand in a dry-weight ratio of 1:2:7. The pH (1N KCl) was adjusted to 6.0 +/- 0.5 with calcium carbonate. The water content was kept at 55% on a dry mass basis. Temperature 15 degrees C. Forty worms were tested at each test concentration.
- Reproduction test (14 days-EC50): Similar above experimental conditions, however, some modification following; a) the replacement of the artificial soil with a natural sandy soil (Kooyenburg) containing 3.7% organic matter, 1.4% clay, and pH (1N KCl) 4.8, and b) the supply of coarsely groud air-dried leaves of alder, Alnus glutinosa, for food.
Measurements/observations
Body weight and number.
Evaluations
14 days-LC50 according to the trimmed Spearman-Karber method (Hamilton et al. 1977). 14 days-EC50 using the statistical software package GENSTAT 5. NOEC applying analysis of variance.
[ref. ID; 1433]
Test system
Worm sampling sites
- (i) The roadside verges of the A660 road in Leeds, West Yorkshire.
- (ii) The roadside verges of the A1 road in West Yorkshire.
- (iii) The verges of a service road through the University of Leeds farm (Spen Lane), at a site 300 m from the site by the A1 road.
- (iv) A city recreational area of deciduous woodland in Leeds (the Ridge), situated near a number of industrial sites.
- (v) A control site at Glen Sheilach, Oban, Scotland.
Measurements
Amounts of cadmium, lead, copper and iron in samples.
[ref. ID; 2167]
Test system
A 32-days toxicity test
Toxicants
Terbufos (COUNTER-15G)
Temperature
8-18 degrees C.
Test design
12-L polyethylene plastic containers. EPA artificial soil (68% silica sand (No. 70 mesh: sand-blasting grade), 20% kaolin clay, 10% peat moss, 2% finely ground calcium carbonate), 12L:12D photoperiod.
Measurements/observations
Mortality, body weight, pesticide uptake.
[ref. ID; 4496]
Test system
60 days toxicity test
Strains
Collected around the Faculty of Sciences in Reims, France, in an area that has not been treated with pesticides for 35 years. Adults with a well-developed clitellum.
Toxicants/concentrations
Isoproturon (0, 0.5, 0.8, 1.0, 1.2, and 1.4 g/kg soil).
Test design
70-90% relative humidity, 12:12 photoperiod. Temperature 14+/-1 degrees C.
Measurements/observations
Body weight, growth rate, mortality, protein contents.
Evaluations
LC50.
[ref. ID; 4498]
Test system
Strains
Clitellate adult (4 to 5 g).
Toxicants
Atrazine-metolachlor liquid formulation.
Test design/concentrations
One hundred-twenty 0.5 L glass jars (200 g moist (13 % w/w) soil (from a zero-till corn field near Belmont, ON, Canada) + concentration (0, a recommended field rate, x3 times, and 6 times), food (2 g soybean leaves, 2 g corn leaves, or no residures).
Measurements/observations
Body weight.
[ref. ID; 4958]
Test system
NRRT assay
Strains
Mature adult individuals were obtained from a commercial supplier (Appats Saint-Gabriel, Saint-Gabriel de Brandon, QC, Canada).
Toxicants
HMX, RDX, TNT, Tetryl.
Test design
Test soil was contaminated with different PNOs such as HMX, RDX, TNT and its by-products.
Measurements/observations
NRRT.
[ref. ID; 5992]
Test system
Heavy metal concentrations in the tissues, ingesta and faeces
Collection sites
Earthworms were collected by formalin extraction (20 L, 0.55%) from five separate 1 m2 sites situated on a longitudinal transect across the Cefn Parc Pb and Zn-mine, South Wales (O.S. grid ref. = ST 048822).
Test design
Earthworms were starved on moistened filter paper for 4 days by which time the alimentary canal was clear of ingested soil material. Animals were wet oxidized with concentrated (16N) "Analar" nitric acid, and analysed for Cd, Cu, Pb, and Zn by flame atomic absorption spectrophotometry.
Measurements/observations
Dry weight and tissue metal concentration of worms, metal concentrations of ingesta (crop contents) and egesta (faeces).
[ref. ID; 5994]
Test system
Effect of urbanization on earthworm community
Study sites
The study was carried out in Brussels, Belgium. A transect was demarcated along heavily trafficked streets running from the city centre to the suburbs, in which a decreasing gradient of urbanization was predicted. Within the transect, six public parks were chosen for the study.
Sampling method
Earthworms were extracted by an electrical octet method (Thielemann, 1986).
Measurements/observations
Average density and biomass of earthworms at individual sites, Pb, Cd, Cu, Zn, Ca, and Mg concentrations of worms.
[ref. ID; 5997]
Test system
The heavy metal concentrations of lead, zinc and cadmium in earthworms and soil from pasture near the Avonmouth smelter (Severnside near Bristol, England)
Samples
Soil and earthworms were taken from two sites. The first site was permanent pasture at Severnside, 4 km from the smelting works. The second site was an apple orchard at Long Ashton at a distance of 9.3 km from the smelter.
Sampling method
The formalin method (Raw, 1959) was used to extract the earthworms from the soil for population estimates and metal determinations. Each worm sample consisted of the number emerging in a 0.25 m2 quadrat and ten samples were taken at each site.
Measurements
Concentration of lead, zinc and cadmium in worms tissues were estimated by atomic absorption spectroscopy.
[ref. ID; 6009]
Test system
Optimized design for earthworm survival test in soil
Strains
The worms had been freshly obtained from a commercial angling-bait supplier in Ontario.
Test design
- Experiment A: [Soil properties] 11 soils (agricultural soils, humus enriched garden soil, boreal forest podzolic sands, and organic) were placed in 500-ml plastic containers with soil surface areas of 90 cm2, respectively. 15 degrees C, 5 mature worms per container.
- Experiment B: Soil-to-worm ratio (80, 40, 33, 27, 20 cm3 soil/worm), type of container (plastic bag, bag + tray, bucket), 15 degrees C.
Measurements
Mortility.
Evaluations
Probability.
[ref. ID; 6073]
Test system
The concentration of volatile chlorinated hydrocarbons in worms sampled from contaminated site and the effect of TCE on worms
Toxicants
cis-1.2.-Dichloroethene, Dichloromethane, 1.1.1.-Trichloroethane, Trichloroethene (TCE), Tetrachloromethane, Trichloroacetic acid (TCA).
Test design
Field study: Earthworms were collected in the northern part of the Black Forest near Pforzheim (west Germany). The worms were collected with electricity using the Octet-Method (Thielemann, 1986).
Laboratory study: 10 Lumbricus terrestris were kept in closed boxes (32x25x10 cm = 8 litres) containing 4 litres of uncontaminated natural soil.
Box 1: 100 ml m-3 TCE, 15 days.
Box 2: 500 ml m-3 TCE, 15 days.
Box 3: 5000 ml m-3 TCE, 9 days.
Box 4: On wet blotting paper without soil, 500 ml m-3 TCE, 15 days.
Control box: 15 days.
Measurements
The concentrations of cis-1.2.-Dichloroethene, Dichloromethane, 1.1.1.-Trichloroethane, Trichloroethene (TCE), Tetrachloromethane, Trichloroacetic acid (TCA) in worms.
[ref. ID; 6075]
Test system
The EDX-measurements on different tissue samples
Toxicants
Sewage sludge (including Chromium, Nickel, Copper, Zinc, Cadmium, and Lead).
Test design
Artificial soil test method.
3 litre test container: Artificial soil + sewage sludge (uncontaminated treatment (control), contaminated treatment) + 3 earthworms. 8 weeks, 10 degrees C.
Measurements
The concentration of heavy metal in tissue of worm using EDX-spectrometer.
[ref. ID; 6078]
Test system
Mercury concentrations in surface soils (0-2 cm), grass (Festuca rubra L.) and earthworms (Lumbricus terrestris L.)
Samples
Within 0.5 km and 10-30 km around a chlor-alkali works (The chlor-alkali process is recognised as a potential industrial emission source of mercury).
Measurements
L. terrestris were killed by deep-freezing and, after thawing, their gut contents removed prior to digestion. Digestions were carried out in Kjeldahl flasks using a concentrated 'Analar' nitric/perchloric acid mixture (4:1). Initial digestion in the cold followed by careful heating was found to prevent any evaporation losses of mercury. Digests were analysed for total mercury with a Varian AA4 atomic absorption spectrophotometer.
[ref. ID; 6101]
Test system
Sublethal toxicity test (Sperm count as a biomarker for environmental toxicology)
Strains
Adult L. terrestris, purchased from Carolina Biological Supply (Burlington, NC).
Toxicants/concentrations
Technical chlordane (6.25, 12.5 and 25 ppm) and cadmium nitrate (100, 200 and 300 ppm).
Test design
The artificial-soil (AS) protocol described by Greene et al. (1989). Adult earthworms, in groups of nearly equivalent masses (4-6 g) were then exposed to chlordane or cadmium mixed with 100 g of AS in 250-ml glass jars, which were maintained in the dark at 10 degrees C within an environmental chamber.
Measurements
Sperm counts, body mass weight.
[ref. ID; 6699]
Test systems
Effect of soil organic matter content for the bioavailability of malathion
Strains
National Association of Supplies Bait and Tackle, Marblechead Ohio.
Toxicants
Malathion (surface exposure 50 ug/cm2).
Test design
- High organic matter content (55%) soil: A commercial Scott's garden soil.
- Low organic matter content (8%) soil: The garden soil : inert silica sand = 1:1 (v:v).
1000 mL glass beaker filled with 600 mL of soil. Malathion was applied by fine mist. Exposures were run in triplicate with three worms replicate, at 10 degrees C, on a 12-hr day/night cycle, during 72 hr.
Measurements/observations
Cholinesterase activities in worm tissue, marathion body burdens.
[ref. ID; 6744]
Test system
Effect of time and mode of depuration on tissue copper concentration
Strains
From Walker Organics, West Creiglee Farm, Roddymoor, Crook, County Durham, UK. Individuals with fully clitellate (mean fresh weights: 1394+/-94 mg).
Toxicant/concentrations
Cu(NO2)3/3H2O, 350 mg Cu kg-1.
Test design
Culture period 28 days.
- Soil: Kettering sandy loam soil is similar in its properties to the OECD standard soil. Moisture content 50%.
- Worm: Eighty earthworms were added to 12 kg of the treated Kettering loam. 15+/-1 degrees C, constant darkness.
Depuration method: 1) Depuration time (0, 24, 36, 48, and 72 hr). After depuration the filter papers (Whatmann no.540) were dried at 40+/-1 degrees C for 24 hr and re-weighed. The earthworms were rinsed in ultrapure deionised water, dried on tissue paper and immersed for 20sec in boiling ultrapure water. 2) After depuration for 0, 24, 36, 48 or 72 hr, the earthworms was dissected following immersion in boiling water for 20sec. The earthworms were slit open to expose the alimentary canal, which was cut from above the crop to the bottom of the intestine. Each earthworm was pinned open onto a wax board and a small brush was used to carefully remove the soil particles present in the crop, gizzard and intestine.
Measurements/observations
The concentration of Cu and Ti in tissues of worm.
[ref. ID; 6748]
Test system
p,p'-DDE bioaccumulation from compost and soil
Strains
From uncontaminated soil.
Toxicants
Soil (56% sand, 36% silt, 8.0% clay, 1.4% organic carbon, and bulk density 1.14 g/cm3) containing p,p'-DDE residues as a result of historical application of DDT was collected from the Conneticut Agricultural Experiment Station's experimental farm. 474+/-59.3 ng p,p'-DDE/g of soil dry wt.
Compost aged approximately 6 months was collected from Lehigh Country, PA, USA. It contained 37.0% organic carbon and had a bulk density of 0.61 g/cm3. The compost contained p,p'-DDE residures. 539+/-79.8 ng p,p'-DDE/g compost dry wt.
Test design
350 g each dried, seived medium (soil and compost) were mixed with 30 g of Perlite added to seven 1000-ml glass beakers (3 replicates + no-worm control). 4 individuals (approximately 9 g biomass) added each beakers. 22+/-2 degrees C in dark.
Measurements/observations
Biomass of worm and p,p'-DDE concentration in the worm tissue.
[ref. ID; 6753]
Test systems
CbE activity in L. terrestris and evaluate its use as a complementary biomaker of anti-ChE pesticide exposure
Strains
A local commercial supplier (Armeria 20, Toledo, Spain), who imported from a commercial vermiculture supplier (Vivastic, Elsenheim, France).
Toxicants
Chlorpyrifos-oxon
Test design
- Run 1: The tissue distribution of total CbE activity using three model substrates (alpha-naphthyl acetate, 4-nitrophenyl valerate and 4-nitrophenyl acetate).
- Run 2: The variations of enzyme activity levels with sexual maturation (presence or absence of clitellum).
- Run 3: The long-term stability of CbE activity.
- Run 4: The tissue-specific differences in CbE isozyme abundance, and the sensitivity of CbEs to chlorpyrifos-oxon inhibition.
Evaluation
IC50.
[ref. ID; 6763]
Test system
Strains
Adult earthworms were provided by Edwin Berry, National Soil Tilth Laboratory, USDA Agricultural Research Service, Ames, Iowa. The earthworms species used were Lumbricus rubellus (weighing 0.80-1.21 g), Aporrectodea trapezoids (weighing 2.15-2.57 g), Lumbricus terrestris (weighing 5.08-6.23 g).
Toxicants
Plasmid: pJP4, Donor: Alcaligenes eutrophus JMP222N, Recipient: Pseudomonas fluorescens C5t
Test design
Soil: A Hubbard loamy sand (Udorthenic Haploboroll).
Microcosm: The microcosm consisted of a polyvinyl chloride tube (50 by 10 cm diameter) which was cut into eight 5-cm segments. The remaining 10-cm segment served as the headspace.
Experimental design
Soil microcosms containing spatially segregated, coinoculated donor and recipient strains, inoculated control (donor and recipient strain only) soil microcosms, and uninoculated control soil microcosms. Triplicate microcosms for each of the four treatment (i.e., no worms, L. rubellus, A. trapezoides, and L. terrestris). 3 adult earthworms added to surface of the microcosm. (The final earthworms density was equivalent to approximately 100 earthworm per m2 of soil. 20 degrees C. Experimental period 2 weeks.
Measurements/observations
Vertical distribution of earthworms number. Donor, recipient, transconjugant bacteria number of casts and cocoon.
[ref. ID; 6770]
Test system
Risk assessment of GEMs
Genetically engineered microorganisms (GEMs)
Pseudomonas fluorescens DSM 50148. It was transformed with plasmid pFL105, conferring resistance to sulfonamides and kanamycin. This chimeric replicon contains RSF1010 and an EcoRI fragment from plasmid K5 carrying the aphB (Km(r)) gene of Tn5 undder control of the CaMV 35S promotor. Plasmid RSF1010 and derivatives have been stably maintained in a variety of Gram-negative bacteria including pseudomonades.
Test design
Plastic column with a diameter of 15 cm and a length of 50 cm was filled up with soil (the Agricultural Research Center at Braunschweig), compressed to a defined bulk density of 1.37 g cm-3 and kept at 12 degrees C. The water content of the soil was 17% (weight:weight; +/-0.5%).
One worm (15 cm length) introduced into column, and then, was fed with 500 ul of a culture of GEMs containing 10E9 cells per ml and 1 g dry food (cow dung).
- After 26 days, earthworm was transferred to a fresh soil column.
- After 51 days, earthworm was again transferred to a fresh soil column.
Measurements/observations
Number of GEMs (CFU) in L. terrestris excreta.
[ref. ID; 6803]
Test system
Food-chain transfer of Cadmium and Zinc
Strains
From a commercial supplier (Blades Biological) mass, 5.38+/-1.25 g, n=50.
Toxicants
Urtica dioica grown for 63 days under hydropoic conditions in Hoagland's solution spiked with Cd (CdSO4/8H2O: 0, 1, 2, 3, and 6 mg Cd/L) or Zn (ZnSO4/7H2O: 0.02 (control), 5, 10, 20 and 50 mg Zn/L).
Test design
Earthworms were placed individually in 0.5-L, black plastic bottles (diameter, 77.0 mm; height 176.5 mm) that had been filled 24 hr earlier with 375 g of air-dried Kettering loam (Broughton Turf and Loam Management) and 125 g of deionized water; the bottles were secured with venting lids. After a further 24 hr, 0.6 g of crushed, air-dried U. dioica leaves were placed on the soil surface of each bottle. The bottles were placed in a randomized block design, with five replicates for each Cd and Zn concentration in the controlled-environment chambers at 15 degrees C and 80% humidity under a 16:8-hr artificial light:dark photoperiod, although the black bottles ensured complete darkness. After 7 days, any uneaten food was removed from the soil surface and replaced with another 0.6 g of the appropriate leaves. This was repeated every 7 days for 28 days.
Measurements
Cd and Zn concentration in the body tissue of worm.
[ref. ID; 6850]
Test system
The use of acid insoluble residue as a marker fraction in the soil
Strains
Lumbricus terrestris, Allolobophora longa, Allolobophora caliginosa, Allolobophora chlorotica were collected from the soil in Rothamsted Park. Eisenia foetida was collected from cattle manure. Mature, clitellate individuals for experimental use.
Toxicants
Zn, Cu, Cd, Pb. Soil sampling site; 4 soils (Frongoch, Ystwyth, Shipham, Broek Polder).
Test design
Groups of each species of earthworm were placed on separate subsamples of each soil. A ratio of approximately 5 g (live weight) Of earthworms to 600 g (air dry weight) of soil. Experimental period 15 days. Temperature 15 degrees C.
- Whole worm: Immediately after removal from the soil, earthworms were rinsed in distilled water, killed by oven drying at 85 degrees C to constant weight. These 'whole worm' samples comprised both earthworm tissue and soil within the alimentary canal.
- Dissected worm: Earthworms were dissected along the entire length of the alimentary canal which was rinsed free of soil with distilled water before being oven dried at 85 degrees C to constant weight.
- Starved worm: Earthworms were placed in clean petri dishes at 100% humidity (a few millilitres of distilled water) for 48 hr (transferred to clean petri dishes after the first 24 hr). All soil egested by the earthworms during the 48 hr period was collected, dried and analysed for acid insoluble residue. Earthworms which had been starved for 48 hr were rinsed in distilled water, killed by oven drying at 85 degrees C to constant weight.
Measurements/observations
Heavy metals and AIR concentration in earthworm tissue.
[ref. ID; 6856]
Test system
Depuration and uptake
Strains
From an Ontario commercial bait supplier. Sexually mature individuals for experiment use.
Toxicants
Iodine-[125], Cs-[134], Mn-[54], Zn-[65] and Cd-[109].
Test design
- Container type 1: 500 ml plastic food containers with small holes in the lid for ventilation, filled with litter.
- Container type 2: Opaque acrylic columns, 5 cm inside diameter by 37 cm deep, filled with a commerical potting soil. These had screen bottoms to allow for free passage of air and water and 500-ml plastic containers fixed to the top to hold the spiked litter.
Litter was collected in the spring in a mixed forest region in southern Manitoba, Canada. A 1-kg wet-weight (69% moisture content) aliquot of litter was spiked, using multiple additions of 10 ml of the spike solution.
Measurements/observations
Weight and Gamma activity of worm.
[ref. ID; 6858]
Test system
Life-cycle and biomarker responses to zinc in four earthworm species (Aporrectodea caliginosa, Eisenia fetida, and Lumbricus rubellus)
Strains
From a commercial supplier. Adult worm.
Toxicants/concentrations
Zn(NO3)2/6H2O aqueous solutions: 0, 190, 350, 620, 1200, 2000, and 3600 ug/g.
Test design
Zinc exposures were conducted in a natural soil-based test system. The soil used was a mixture of a commercially available sandy loam soil (Rockalls, Wokingham, Berkshire, UK) and commercially available Sphagnum peat (Bullrush Ltd, County Tyrone, UK). One kilogram of the soil mix was added to each experimental a container (plastic boxes 220x160x80 mm), with four replicate containers used for each test concentration. Water-holding capacity 60%. 6 worms per container. During exposure period, the test containers were covered to limit water loss and kept in constant light at 15 degrees C for 42 days. Finely ground fresh horse manure (dried and rewetted to 75% water content) was added as source of food (4 g dry weight per weekly to each container) in all tests.
Measurements/observations
- Life-cycle parameter: Survival, weight change, and cocoon production rate.
- Biomaker: Neutral-red retention by coelomocytes lysosomes.
- Zinc conconcentration in worm tissues.
Evaluations
Significant differences in parameters were calculated using analysis of variance (ANOVA). When, differences were found, Tukey's multiple comparison test was used to determine differences between specific treatments.
- LC50 by the log-probit method using the MicroProbit 3.0 statistical software package.
- EC50 using the linear interpolation technique within the ICp 2.0 software system.
- Sublethal sensitivity index (SSI) were determined from calculated effect concentration (EC10) values. SSI=LC50/EC10.
[ref. ID; 6894]
Test system
The role of middens formed by Lumbricus terrestris
Sampling sites
The samples were collected from a silt loam soil under a corn (Zea mays L.)-soybean (Glycine max L.) rotation in west-central Indiana U.S.A.
Toxicants
[14]C-Atrazine.
[ref. ID; 6897]
Test system
To standardize the phagocytic activity assay in coelomocytes using flow cytometry
Strains
From a commercial supplier (St-Eustache, Quebec, Canada).
[ref. ID; 6899]
Test system
Applications of toxicity curves and incipient lethal level (ILL) in assessing the toxicity of soil contaminant
Strains
From commercial suppliers.
Toxicants
Diazinon, PCP.
Test design
3 natural soil (Brookston Clay, Agriculture Canada Harrow Research Station; Guelph Loam, University of Guelph Arkell Research Station; Fox Sand, Agriculture Canada Delphi Research Station). The collection sites had known land-use histories and were considered to be free of pesticide contamination.
Test soils (487.5 g dry wt) were weighed into 2-L clear glass jar. Water-holding capacity 80%. 10 worms per jars. Exposure period 21 days. 3 replicates.
Measurements/observations
Mortality (2, 4, 8, 16, 24, 48 and 96 hr, 5, 7, 14 and 21 days).
Evaluations
- LC50 by trimmed Spearman-Karber analysis.
- ILLs were estimated by fitting a one-comparment first-order kinetics model to the LC50/time data using non-linear regression analysis.
[ref. ID; 6902]
Test system
Strains
From bait shops and acclimated in the laboratory for four months.
Toxicants
Parathion (Formulation E 605 forte Bayer, 50% a.i.) and Amitrole-Diuron (Ustinex).
Test design
OECD-Guideline 207 (OECD, 1984). Experimental period 28 days.
Soil substrate (artificial soil or loamy natural soil (5.6% sand, 75% silt, 19% clay; 2.1% organic substance; pH 6.0; air-dried and sieved)), temperature (20 or 10 degrees C), soil moisture (82% or 51% Max. water capacity), way of application (mixing pesticide into the soil or spraying onto the soil surface).
1000 g (dry wt) soil and 3 worms per test container. Each container was supplied weekly with 2.5 g (dry wt) of a mixture of two-thirds air-dried, finely ground cow manure and one-third dried leaves of stinging nettle (Urtica dioica L.).
Measurements/observations
Mortality, weight, cocoon production, and behavioural changes.
[ref. ID; 6903]
Test system
Sampling sites
Earthworm population collected from four sites: the median strip of a busy street in Cracow; a city park (100 m from the busy street); a forest about 30 km from Cracow (100 m from a low-traffic local road); a forest on calcareous soil about 30 km from Cracow (distant from traffic).
Toxicants
Traffic pollution (Pb).
Measurements/observations
Species composition, population density and biomass of the earthworms.
Lumbricus terrestris were analyzed Pb concentration in tissue and haemoglobin content.
Evaluations
Data were subjected to one way ANOVA with TUkey HSD range test, or Kruskal-Wallis one-way analysis by ranks and Spearman Rank Correlations (Sokal and Rohlf, 1981).
[ref. ID; 6904]
Test system
Avoidance test
Strains
Adult worms, approximetly 4500 mg having a well developed clitellum (more than 6 weeks old).
Toxicants
Diazinon, Sevin, Safer Fruit and Vegetable Insect Killer, Lindane, Malathion, Orthene, Spectracide, Captan, Metaldehyde, Daconil.
Test design
A double layer of glass spheres, approximately 1.25 cm dia, underlying a wire mesh screen barrier, was placed into glass test chambers (30.5x25.4x5.1 cm). Each chamber was then divided into two sections of equal size using a glass divider. Soil mixture for each section of the test chamber was prepared by combining 190 g fine, synthetic potting soil (Hoffman Seed Starter containing vermiculite, peat moss, and limestone), 14 g dried, pulverized leaves, and 37 ml distilled water. Each pesticide was added to and blended thoroughly with the prepared soil mixture. 5 worms were added to each pesticide-soil mixture, and the earthworm-pesticide-soil combination was placed in one side of each test chamber. The test chambers were held in a dark room maintained at 13 degrees C for 48 hr.
[ref. ID; 6905]
Test system
Acute lethality
Strains
Toxicants
Diazinon (Basudin R 500 EC).
Test design
- Laboratory experiment: 2-l glass test container (including Fox sand, water and diazinon) were randomly placed onto a table in a room with a constant temperature (17+/-1 degrees C, and the fluorescent lights were kept on to ensure that the worms penetrated the soil. 15 test containers per treatment (diazinon concentration 10, 40, 80 and 160 mg diazinon kg-1 soil dry wt, 3 replicates each concentration). 10 worms per container. Sampling time is day 4, 7, 14 and 21.
- Field experiment: Test containers were made with flexible corrugated drainage tiles with vinyl fly screens fixed over both ends. The test containers with soil (750 g of Fox sand) and 10 worms were randomly placed into prepared holes in outside plots, where they would be exposed to natural climatic conditions. The top of each test container was fitted with a 7.4 cm dia plug of bluegrass sod. Spray applications of diazinon (0.5, 1.0 or 2.0 times the application rate of 7.5 kg a.i. ha-1) in distilled water were applied to the surface of each test container (12 test containers per treatment). Sampling time is day 4, 7, 14 and 21.
Measurements
Mortality. Diazinon concentrations in worms.
Evaluations
7-days and 21-days LC50, BCF.
[ref. ID; 6951]
Test system
Acute toxicity (7 and 14 days)
Strains
Strains supplied by Wolkins Bug House, Martinsville, Indiana.
Toxicant/concentrations
Benomyl (0.0, 0.03, 0.1, 0.3, 1.0, and 3.0 ppm).
Test design
- 1. Effect of test media (slurry): Nonsterile sandy loam soil 900, 890, 875, 870, 850, 825, 800 g + food (rabbit feces) 0, 10, 25, 30, 50, 75, 100 g + water 100 ml.
Approximately one-half of the test medium was placed in the test containers (2-liters glass battery jars). The worms placed onto the test medium, and covered with the remaining test medium. The test medium was covered with a piece of moistened cheesecloth and held in the dark at approximately 13 degrees C.
- 2. Toxicity test: Test medium soil 850 g + food 50 g + water 100 ml, 5 worms, triplicate.
Measurements
Mortality and body weight.
Evaluations
LC50.
[ref. ID; 6952]
Test system
LC50 and LD50
Strains
From local bait shop. Sexually mature worms weighing 4-5 g with well developed clitella were used.
Toxicants
Carbaryl, DDT, solvent (acetone, acetonitrile).
Test design
13 degrees c in the dark. Exposure period 1, 5, 7, 14, 21 days.
- 1. LC50: Dips were performed by placing 10 ml of each concentration of compound in a 50-ml beaker. Worms were added individually to beakers and exposed for 10 or 30 min. Upon removal, the worms were placed in jars containing 100 g of untreated standard assay soil described by Karnak and Hamelink (1982) (unsterilized potting soil/distilled water/ground rabbit feces, 7:2:1). For soil incorporation, following changes: 1 ml of each concentration of compound, dissolved in acetone, was added to 200 g of standard assay medium and mixed thoroughly; five worms were exposed per jar. Six replicates.
- 2. LD50: Topical application was accomplished using an Instrumentation Specialites microapplicator with a 1.0-ml Hamilton Syringe fitted with a 27-gauge needle. Worms to be injected were anesthetized by placement on ice 15 min prior to dosing. Ethanolic solutions of pesticide were injected in volumes of 1-2 ul into the hemocoel directly behind the clitellum using a microapplicator.
Measurements/observations
Weight loss, appearance of sores or swellings, coiling, and mortality.
Evaluations
LD and LC50 values were determined according to the method of Finney (1971).
[ref. ID; 7014]
Test system
Avoidance test
Strains
From a local supplier.
Toxicants/concentrations
Brass powder (70% Cu, 30% Zn): 0, 17, 38, 120, 200 ug/g.
Test design
15 cm x 50 cm styrofoam chambers filled with nonsterile loam soil (600 g), distilled water (150 ml), and then the brass powder spike. The soil spiked with the brass powder was placed in one-half of the chamber and uncontaminated soil was placed in the other half. Five earthworms were added at the interface between the two soils. The cambers were placed randomly into a precision low-temperature incubator set at 13.0+/-0.2 degrees C. Exposure period 7 days.
Measurements/observations
The locations of the earthworms in the chamber.
Evaluations
The threshold avoidance concentration.
[ref. ID; 7015]
Test system
LC/LD50 (5 days) and Immunoassay
Strains
From a commercial biological supply company.
Toxicants
Aroclor 1254 (hereon PCB)
Test design
- Filter paper experiment: PCB concentrations (2.5, 5.0, 10.0, 40.0, 50.0, 100, 200, 400 and 800 ug/cm2) on Whatman #1 filter paper disks (9 cm diameter) in 0.47-liter glass jars. The jars were sealed with lids and placed into environmental chambers without light at 10+/-1 degrees C for 5 days.
- Uptake/depuration experiment: Earthworms were exposed PCB concentration of 10.0 ug/cm2 for 5 days and then returned to containers of peat moss for depuration. During exposure, sample earthworms were removed at 1, 4, 8, 16, 32, 64 and 120 hr. Depuration samples were weekly during 5 months.
- Immunoassay: During uptake and depuration phases, coelomic leukocytes from experimental and control earthworms (N=14-20 assay) were obtained by a noninvasive extrusion protocol.
Measurements/observations
- Filter paper method: Mortality and PCB concentration in worm entire body.
- Uptake/depuration experiment: PCB levels in carcass tissue (all tissue except coelomic contents), and coelomic leukocytes and fluid.
- Immunoassay: Number of secretory rosettes and erythrocytic rosettes formed by 100 randomly counted coelomic leukocytes.
Evaluations
Filter paper method: LC/LD50 by probit anaylsis.
[ref. ID; 7016]
Test system
On-site method for assessing chemicals
Strains
From a local bait dealer.
Toxicant
Site: The Baird and McGuire Superfund Site, Holbrook, Massachusetts, a former site for mixing pesticide batches for resale, is a mixed upland/wetland forested area near Boston, Massachusetts.
Test design
The bioassay chambers were laid out in transects the area of suspected contamination and in a reference area. Field survey methods (distance and bearing) were used to tie the transect into known survey points (accuracy +/- 1.5 m). Plastic chambers (approximately 4-L) were placed into the soil by excavating soil to a depth of about 20 cm, which filled the container to about three-quarter capacity. The cambers permitted free flow water and air. Five L. terrestris were placed on the surface of soil, and then the camber was with cheesecloth cover secured with a rubber band. Exposure period 7 days.
Measurements/observations
Morbidity (burrowing, coiling, shortening, stiffening, swelling and lesions) and Mortality.
Nine chemicals (alpha-chlordane, Chlordene, Endrin, gamma-chlordane, gamma-chlordene, p-DDD, p-DDE, p-DDT, Nonachlor) concentrations in worm tissue and soil.
Evaluations
- RANK 4 = death of oe to five worms.
- RANK 3 = any occurrence of lesions without death.
- RANK 2 = a combination of the sublethal endpoints.
- RANk 1 = no obvious impact of nay kind.
[ref. ID; 7020]
Test system
24-hr Allogeneic cytotoxicity assay
Strains
From Golden West Cricket Co. (Paramount, CA) and Carolina Biological Suplly Co. (Burlington, NC).
Toxicants
Aroclor 1254.
Test design
Single pieces of 9-cm Whatman no.1 filter paper were placed flat on the bottoms of glass jars. 1 ml Aroclor 1254 (10 ug/cm2 paper) in acetone or acetone only (controls) was dropped onto the paper, wetting the entire surface. Acetone was allowed to evaporate overnight, 1 ml water was dropped onto the paper, and one worm was placed in each jar. Lids were adjusted loosely onto jars and then transferred to 15 degrees C incubator for 48 hr.
Measurements/observations
Cytotoxicity assay (trypan blue assay) and Lactate dehydrogenase assay by using coelomocytes.
[ref. ID; 7026]
Test system
The effect of Edaphic factor (soil organic matter content, pH, and depth in soil profile) affecting toxicity and accumulation
Strains
From a bait shop.
Toxicants
Disodium Arsenate.
Test design
Soil (a beech-Scotch pine canopy, UK. Arsenic level 1.2 ug/g).
- Experiment 1: Arsenate-dosed soil (400 g dry weight, 0-70 cm depth in soil profile) was placed in 500-ml Kilner jars. The tops of the jars were covered with clear polyethylene punched with holes. 5 worms in each jars. 3 replicates. Arsenate doses 0, 15, 30, 40, 50, 60, 70, 80, 100, 200, 400 and 500 ug/g. Exposure period 10 days.
- Experiment 2: Soil from following depth 70-90, 90-140, 140-300, 300-500, and 500-700 cm. Arsenate doses 0, 50, 100, 200, 300, and 600 ug/g. Exposure period 4 days.
- Experiment 3: Arsenate doses 40 ug/g dry weight. Exposure period 23 days.
Measurements/observations
Mortality and arsenic concentration in worm tissues.
Evaluations
LC50.
[ref. ID; 7084]
Test system
The potential for trophic transfer
Strains
From Carolina Biological Supply Company.
Toxicants
ZnCl2 (Zn2+: 4.7, 22.9, 52.7, 106.6, 130.7, 173, 259.6 275.8, 287.8, 372.5, 399.9, and 476.3 mg/kg)
Test design
Three earthworm were weighted and placed into each microcosm containing 300g of OECD artificial soil. pH approximately 6. Each microcosm was placed inside an incubator and held at 70% humidity and 16+/-3 degrees C on a 12 hr dark:light cycle. Exposure period 21 days.
Measurements/observations
Mortality, body weight, Zn concentration in worm (gut tissue, body tissue).
[ref. ID; 7133]
Test system
Bioavailability
Strains
Isotopic enrichment of worms.
Earthworms were equilibrated in [68]Zn-spiked backgroud soil (the outer perimeter of a storm-water pond basin in Owings Mills, Maryland, USA).
Toxicants
ZnCl2 only (0, 25, 50, 75, 100, 125, 150, and 175 mg/kg Zn). ZnCl2 (175 mg/kg Zn) + CaCl2 (0.1 M), or NaCl (0.04, 0.02, 0.017, 0.013, 0.0075 M)
Test design
One worm was placed in a plastic cup containing 300 g soil wet weight. Fifteen treatments were performed, with three replicate cups each. The worms were kept in a temperature-controlled incubator on a 16:8 light:dark cycle at 16.5 degrees C; humidity at approximately 60%. Exposure period 2 days.
Measurements/observations
[68]Zn/[66]Zn in worm of body tissues, gut tissues, organ tissues.
[ref. ID; 7173]
Test system
Biotransformation
Strains
- 1. Field-collected samples: Adult L. terrestris (clitellum present) were hand collected at the Devon Great Consols (DGC) former arsenic mine, UK, at former gold mine in Deloro, Ontario, Canada and at uncontaminated garden soil in Nottingham, UK.
- 2. Mesocosm samples: L. terrestris for use in laboratory mesocosm exposures were sourced from a commercial supplier.
Toxicants
As.
Test design
Mesocosm experiments: A contaminated bulk soil sample collected at DGC was thoroughly mixed with equal proportions of uncontaminated, unsterilized top-soil and John Innes No.1 compost in increasing amounts to give six soil mixtures (~5 kg) of increasing arsenic contamination. A 1:1 mixture of uncontaminated topsoil and compost was used as the control and contained 1.0 mg kg-1. Depurated L. terrestris were exposed to increasing soil As concentrations (1.0, 98, 183, 236, 324 and 436 mg kg-1) for a total period of 35 days. pH 5.9+/-0.4, moisture 31+/-1.3%.
Measurements/observations
These following material concentration in worm tissues, soil, and litter: Inorganic arsenite (As(III)), arsenate (As(V)), methylarsonate, dimethylarsinate, arsenobetaine and arsenoholine, trimethylarsineoxide and tetramethylarsonium ion, and four arsenosugar compounds (glycerol, phosphate, sulphonate, sulphate).
[ref. ID; 7626 (in German)]
Test system
The chloragocyte-eleocyte transformation
Strains
Toxicants
Benomyl, Carbofuran
Test design
Filter paper method.
Measurements/observations
Electronmicroscopical methods.