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At least 343 records · Page 19Linked to original sources

PubMed2005

Age-specific immunocompetence of the earthworm Eisenia andrei: exposure to methylmercury chloride.

Little information with regard to the sensitivity of the immune system of earthworms to toxicants is currently available. To that effect, we evaluated the yield of coelomocyte immune cells and their phagocytosis potential for four different stages of development of the earthworm Eisenia andrei both for in vitro and in vivo exposure. Cell viability was similar among size classes; extruded cell yield increased from 0.7 x 10(6) cells/worm in the smallest size class to 2.9 x 10(6) cells/worm in the largest size class. The proportion of phagocytically active cells was lower in the smallest groups but not different among the other three groups. Phagocytosis increased incrementally for the first three age/weight groups. The absolute phagocytosis level of the hatchling worms is significantly lower than that of the adult worms. The in vivo filter paper exposure of the mature worms showed a hormetic stimulation of 200-300% of phagocytosis and no significant effects on the phagocytosis potential of the hatchlings. The absolute response of coelomocytes from younger worms was significantly lower and the estimates of lethal mercury concentrations showed the hatchling worms to be roughly three times more sensitive than mature worms.

Age Factors↗

PubMed2005

Species-specific heavy metal accumulation patterns of earthworms on a floodplain in Japan.

We identified all earthworm species found on a floodplain contaminated by heavy metals (Cu, Zn, Cd, and Pb) from an old mine in central Japan and compared their abundance, biomass, and heavy metal concentrations in tissue. There were six species belonging to three families: Megascolecidae, Moniligastridae, and Lumbricidae. Earthworm community structure seemed to be influenced mostly by soil properties, especially pH and clay fraction. Despite the same endogeic characteristics, species-specific patterns of heavy metal accumulation were observed: species in Megascolecidae and Lumbricidae had relatively lower concentrations compared to those in Moniligastridae. Within Moniligastridae, Drawida sp. accumulated Cu and Pb markedly higher than Drawida japonica. Based on heavy metal concentrations in extracts of CaCl(2) and diethylenetriaminepentaacetic acid, the aging caused remarkably low concentrations in pore water, indicating low availability by dermal uptake. Therefore the different patterns of heavy metal accumulation among species would partly result from species-specific gut process.

Animals↗

PubMed2006

Earthworm expulsion by formalin has severe and lasting side effects on soil biota and plants.

The application of formalin to the ground is a commonly used method to estimate the abundance and biomass of earthworms in soil. From April to September 2003, we investigated potential negative side effects of this method on soil organisms and plants in the sampling soil area to which formalin has been applied. The application of 0.4% formalin (40 L per 0.25 m(2)) severely decreased abundances of Collembola and Nematoda, soil respiration, dehydrogenase activity, fatty acid contents, vegetation cover, and shoot and root biomass. These effects lasted for the whole study period. The impact of formalin may have been aggravated due to unusually hot and dry weather conditions. However, owing to its harmful consequences, the formalin method cannot be recommended as a standard for earthworm extraction.

Animals↗

PubMed2006

The determination of earthworm species sensitivity differences to cadmium genotoxicity using the comet assay.

The concept of species sensitivity differences is important in ecotoxicology and environmental risk assessment, but testing usually focuses on lethality of toxicants. The effects on the suborganismal level are mostly ignored; therefore, the present study assessed a biomarker of genotoxicity (the alkaline comet assay) to compare species sensitivities. Five earthworm species (Amynthas diffringens, Aporrectodea caliginosa, Dendrodrilus rubidus, Eisenia fetida and Microchaetus benhami) were exposed for 48 h to sublethal concentrations of cadmium sulphate in reconstituted soil water and DNA integrity was evaluated with the parameter Tail DNA %. Significant amounts of DNA damage were detected in three (A. caliginosa, D. rubidus and E. fetida) species. E. fetida exhibited the highest level of DNA damage, although D. rubidus showed the highest increase (3-fold) in DNA damage from the control. All exposed earthworms accumulated Cd, although body loads did not correspond with DNA damage levels; most of the Cd was probably sequestrated and rendered harmless.

Animals↗

PubMed2004

Profiles of enzymatic activity in earthworms from zinc, lead and cadmium polluted areas near Olkusz (Poland).

The aim of the study was to determine whether there are signs of adaptation of soil fauna to a gradient of heavy metal contamination. Earthworms Aporrectodea caliginosa, Lumbricus terrestris and Eisenia fetida were collected during the spring and summer of 2000 and 2001 from meadow sites situated between 2 and 32 km from the Bukowno-Olkusz complex of zinc-lead ore mines and smelters. The heavy metal content in the soil near smelters reaches 10,500 mg/kg (d.w.) for Zn, 2600 mg/kg for Pb and 81.9 mg/kg for Cd. The sites differ with respect to species composition of earthworm community, with A. caliginosa being dominant. Complete data was obtained only for A. caliginosa, since other species were not abundant at all investigated sites during the whole period of investigation. The body burdens of Zn, Pb, Cd and Cu in A. caliginosa reached 1500, 100, 220 and 10 microg/g, respectively, in the vicinity of the smelter (2-4 km), and decreased to 400, 2, 36 and 6 microg/g at the most distant site (32 km). Cadmium and lead content was significantly elevated in the whole body of L. terrestris collected at the site 2.5 km distant from the smelters when compared to more distant sites, while in E. fetida only the body burden of cadmium was elevated at the nearest site compared to the next site of transect. Activities of glutathione peroxidase (GPX; EC 1.11.1.9) against hydrogen peroxide (H2O2) or cumene hydroperoxide (cumOOH), glutathione reductase (GR; EC 1.6.4.2), glutathione S-transferase (GST; EC 2.5.1.18) and catalase (CAT; EC 1.11.1.6) were assayed in postmitochondrial supernatant obtained from whole body homogenates. Seasonal and annual variations of enzyme activity were reflected by higher GPX activity in the late summer of 2001 in comparison with the spring and summer of 2000. This may reflect severe drought in the spring and summer of 2000. The activity of both GPX isozymes, GR and GST in A. caliginosa and L. terrestris increased with increasing distance from the smelters and reached maximum at sites III and IV (4 and 8 km from the smelters, respectively) and then it decreased in the animals from site V (32 km). These may be the effects of antagonism between the enzyme inducing and enzyme inhibiting action of smelter emissions, a phenomenon known as a hormetic effect. It is postulated here that this effect is of diagnostic value for metal pollution biomonitoring.

Acetylcholinesterase↗

PubMed2004

Biomarker responses of the earthworm Aporrectodea tuberculata to copper and zinc exposure: differences between populations with and without earlier metal exposure.

Biomarkers in the earthworm Aporrectodea tuberculata (Eisen) were measured to find out their possible induction under Cu and Zn exposure and differences in the responses between two populations with different exposure history. The biomarkers applied were concentration of metallothioneins (MT), and cytochrome P4501A (CYP1A) monooxygenase and glutathione-S-transferase (GST) activities. These were measured from earthworms sampled at three distances from a steel smelter in Finland and from the individuals from two populations, one with and another without earlier metal exposure, exposed to three combined Cu/Zn concentrations in the laboratory. In the field, MT concentration, and cytochrome CYP1A and GST activities decreased with increasing distance from the smelter. In the laboratory, biomarker responses varied in relation to the duration and level of exposure, and they were also dependent on the population. The endpoints appeared to be analytically reproducible and sensitive parameters in A. tuberculata, and thus, they can be used in ecotoxicological field monitoring and in experimental research in the laboratory.

Animals↗

PubMed2005

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

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

Animals↗

PubMed2006

Metal accumulation in the earthworm Lumbricus rubellus. Model predictions compared to field data.

The mechanistic bioaccumulation model OMEGA (Optimal Modeling for Ecotoxicological Applications) is used to estimate accumulation of zinc (Zn), copper (Cu), cadmium (Cd) and lead (Pb) in the earthworm Lumbricus rubellus. Our validation to field accumulation data shows that the model accurately predicts internal cadmium concentrations. In addition, our results show that internal metal concentrations in the earthworm are less than linearly (slope<1) related to the total concentration in soil, while risk assessment procedures often assume the biota-soil accumulation factor (BSAF) to be constant. Although predicted internal concentrations of all metals are generally within a factor 5 compared to field data, incorporation of regulation in the model is necessary to improve predictability of the essential metals such as zinc and copper.

Animals↗

PubMed2006

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

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

Animals↗

PubMed2006

DNA damage and repair process in earthworm after in-vivo and in vitro exposure to soils irrigated by wastewaters.

In this study, DNA damage to earthworms (Eisenia fetida) after in vivo exposure to contaminated soils was measured by detecting DNA strand breakages (DSBs) and causality was analyzed through fractionation based bioassays. A non-linear dose-response relationship existed between DNA damage and total soil PAHs levels. DNA damage, measured with the comet assay, and its repair process, were observed. To identify the chemical causality, an in vitro comet assay using coelomocytes was subsequently performed on the fractionated organic extracts from soils. The results showed that the PAHs in the soils were responsible for the exerting genotoxic effects on earthworms. When normalized to benzo(a)pyrene toxic equivalent (TEQ(BaP)), the saturation dose in the dose-response curve was about 10ng TEQ(BaP) g(-1) soil (dw).

Animals↗

PubMed2006

Glycosylated trypsin-like proteases from earthworm Eisenia fetida.

Although groups of earthworm proteases have been found by several laboratories, it is still unclear how many of the isolated trypsin-like fibrinolytic enzymes are in glycosylated form. Here, eight glycosylated fibrinolytic proteases (EfP-0-1, EfP-0-2, EfP-I-1, EfP-I-2, EfP-II-1, EfP-II-2, EfP-III-1 and EfP-III-2) were isolated from an earthworm species (Eisenia fetida) through a stepwise-purification procedure: ammonium sulfate precipitation, affinity chromatography on a Sepharose-4B column coupled with soybean trypsin inhibitor (SBTI), and ionic chromatography with a DEAE-Cellulose-52 column. Among the eight purified trypsin-like glyco-proteases, EfP-0-2 and EfP-II-2 were newly isolated isozymes. Glycoprotein staining of the proteases on native-PAGE with a Schiff's reagent (sodium meta-periodate) revealed that the eight proteases were glycoproteins. Measurements of the glycan content with sodium meta-periodate and glycoprotein-test reagent showed that these proteases had different carbohydrate contents. Dot-blotting assay with ConA suggested the oligosaccharides were composed of mannose residues.

Amino Acid Sequence↗

PubMedDate not supplied

Evaluation of an innate immune reaction to parasites in earthworms.

Encapsulation is an essential process of the invertebrate immune system and includes the prophenoloxidase (proPO) cascade. We present an assay for evaluating this immune response, now newly adapted to earthworms. Coelomic fluid is withdrawn and coelomocytes are stained with l-Dopa. We studied assay repeatability and the correlation between number of PO-active cells and infection level of the parasitic protozoan Monocystis sp. in the earthworm Lumbricus terrestris. Our study showed high assay repeatability although the expected negative relationship between PO-active coelomocytes and parasite load was not observed; yet a suggestion toward a positive relationship was detected. This finding is contrary to previous assumptions that presume coelomocyte concentrations to be the independent variable determining parasite load.

Animals↗

PubMed2005

Influence of mRNA and protein synthesis inhibitors on the long-term memory acquisition of classically conditioned earthworms.

We investigated the process of memory consolidation following classical conditioning of earthworms. Earthworms were conditioned in paired trials by a weak vibration as a conditioned stimulus (CS), and by light as an unconditioned stimulus (US). The occurrence of a shrinking response upon exposure to the CS increased steadily with the number of paired training trials. When the training procedure was changed by increasing the intertrial interval (ITI), it was found that only those worms trained with a 68 s ITI exhibited long-term memory retention for at least 24 h. The influence of mRNA synthesis inhibition by actinomycin-D or of protein synthesis by anisomycin on memory consolidation was also examined. Induction of the long-term memory was blocked when either of these two compounds was injected into the body cavity of the worm within 25 min of conditioning with the 68 s ITI. These results demonstrate that the long-term memory is dependent upon protein synthesis in response to the upregulation of new transcription messengers.

Animals↗

PubMed2005

Cloning and expression of earthworm fibrinolytic enzyme PM(246) in Pichia pastoris.

We have cloned, expressed, and purified a novel earthworm fibrinolytic enzyme (EFE) of Lumbricus rubellus in Pichia pastoris. Its cDNA sequence revealed a 747bp region containing an intact ORF that encodes a protein of 246 amino acid residues, designated as EFE PM(246). While EFE PM(246) is distinct, its cDNA shows a high degree of sequence homologies with four other EFE cDNAs registered in GenBank. The recombinant EFE PM(246) was active, showing a fibrinolytic activity of 7.5 x 10(6)U/L in basal salts medium, a higher fibrinolytic activity than those produced in other expression systems. The recombinant EFE PM(246) expressed in basal salts medium was purified by a three-step purification procedure with a recovery rate of about 20%. This is the first report detailing the successful purification of a genetically engineered earthworm fibrinolytic enzyme. The main physiochemical features of the EFE PM(246), including temperature stability, pH resistance, and sensitivity to some protein inhibitors, were also characterized.

Amino Acid Sequence↗

PubMed2005

Surface adsorption of metals onto the earthworm Lumbricus rubellus and the isopod Porcellio scaber is negligible compared to absorption in the body.

In terrestrial organisms, bioaccumulation is usually based on a summation of the amount of metal adsorbed to the body wall and absorbed into the body. The relative proportions of metal adsorption and absorption are usually not quantified. In this study, the distinction between adsorbed and absorbed metals was investigated in two different terrestrial species exposed to metals for 2 weeks. The earthworm Lumbricus rubellus was chosen as representative for organisms mainly taking up metals via the dermal route, and the isopod Porcellio scaber as an organism taking up metals mainly via the alimentary tract. Cross-sections of whole animals were made using a cryostat and accumulated metals were localized by means of a phosphor screen (autoradiography). Radiolabels were used to determine the distribution of metals over the different organs and to distinguish between adsorption and absorption. Cd in the earthworm was mainly found in tissues of the chloragogenous region, whereas Zn was also found in various other organs and in the connective tissue. In the isopod, both Cd and Zn were mainly located in the hepatopancreas. Adsorbed amounts of Cd and Zn were negligible compared to internalized Cd and Zn concentrations for both organisms. Consequently, when focusing on effects of metal uptake for the organism itself, there is no need to correct for adsorption. This suggests that adsorption to the epidermis is not a rate limiting step in metal uptake by soil invertebrates.

Adsorption↗

PubMed2006

Formation of filopodia in earthworm (Lumbricus terrestris) coelomocytes in response to osmotic stress.

Coelomocytes, the immunodefense cells of the earthworm Lumbricus terrestris, are exposed to changing osmotic pressures as the worm's coelomic fluid responds to fluctuating wet-dry conditions of the surrounding soil. Using light and fluorescence microscopy combined with actin and tubulin disrupting drugs, we determined the effects of changing osmotic pressure on coelomocyte morphology. The coelomocytes from L. terrestris respond to an increase in environmental osmotic pressure from isotonic conditions (170 mOsm) to hypertonic conditions (715 mOsm) by changing from a round/petalloid morphology to a filopodial morphology. Cytoskeletal fluorescent staining studies indicate that for filopodia to form, the actin cortical ring, present in most coelomocytes in isotonic conditions, must be disrupted. Breakdown of the actin ring by exposure to a hypertonic environment or actin disrupting drugs allows the formation of actin or tubulin-based filopodia. The filopodia, or podial-like extensions formed by earthworm coelomocytes, may enable the cells to better explore their environment.

Actins↗

PubMed1997

Octopamine-containing neurons in the alimentary tract of the earthworm (Eisenia fetida).

Octopamine-containing nerve cells have been demonstrated in the enteric plexus of the earthworm (Eisenia fetida), applying immunocytochemistry and HPLC assay. A few octopamine-immunoreactive neurons occurred in the fore- and hindgut, whereas their number in the midgut was considerably greater. Octopamine levels detected by HPLC correlated with the distribution of octopamine-containing nerve cells. A regulatory role for these intrinsic octopaminergic neurons is suggested in the enteric plexus in the earthworm alimentary tract. This is the first report on the occurrence of octopamine-containing nerve cells in the peripheral nervous system of an invertebrate.

Animals↗

PubMed2001

An NMR-based metabonomic approach to the investigation of coelomic fluid biochemistry in earthworms under toxic stress.

The endogenous metabolites of the coelomic fluid of the earthworm Eisenia veneta were characterised using high-resolution one-dimensional and two-dimensional 1H nuclear magnetic resonance spectroscopy. Signals from common organic acids, such as acetate, fumarate, malonate, malate, formate, and succinate, were identified together with adenosine and nicotinamide mononucleotide. The potential use of this information as a baseline dataset for future toxicological or physiological studies was demonstrated by a metabonomic analysis: a series of earthworms were dosed with the model compound 3-fluoro-4-nitrophenol, and toxic effects followed by multivariate analysis of the spectral data of the coelomic fluid. Relative concentrations of acetate and malonate were decreased in the dosed worms compared to the controls.

Acetates↗