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PubMed2006

Bioaccumulation of heavy metals in the earthworms Lumbricus rubellus and Aporrectodea caliginosa in relation to total and available metal concentrations in field soils.

The aim of this study was to determine important metal pools for bioaccumulation by the earthworms Lumbricus rubellus and Aporrectodea caliginosa in soils with high binding capacity. Cd, Cu and Zn concentrations in soil, pore water and CaCl(2) extracts of soil, in leaves of the plant species Urtica dioica and in earthworms were determined at 15 field sites constituting a gradient in metal pollution. Variations in the Cu and Cd concentrations in L. rubellus and Cu concentrations in A. caliginosa were best explained by total soil concentrations, while variation in Cd concentration in A. caliginosa was best explained by pore water concentrations. Zn concentrations in L. rubellus and A. caliginosa were not significantly correlated to any determined variable. It is concluded that despite low availability, earthworms in floodplain soils contain elevated concentrations of Cu and Cd, suggesting that uptake takes place not only from the soluble metal concentrations.

Animal Feed↗

PubMed2006

Phoresy of the entomopathogenic nematode Steinernema feltiae by the earthworm Eisenia fetida.

The free-living stage of entomopathogenic nematodes occurs in soil, and is an environmental-friendly alternative for biological control. However, their dispersal capability is limited. Earthworms improve soil characteristics, changing soil structure and influencing many edaphic organisms. Thus, earthworms could be used as vectors to introduce/disperse beneficial organisms. Nevertheless this interaction has not been studied in detail. This study presents the infectivity results of Steinernema feltiae after passing through the Eisenia fetida gut. Although entomopathogenic nematodes have no deleterious effects on earthworms, their passage through E. fetida gut seriously affected their mobility and virulence.

Animals↗

PubMed2005

Crystal structure of earthworm fibrinolytic enzyme component B: a novel, glycosylated two-chained trypsin.

The earthworm fibrinolytic enzyme (EFE), belonging to a group of serine proteases with strong fibrinolytic activity, has been used in a mixture as an oral drug for prevention and treatment of thrombosis in East Asia. The EFE component b (EFE-b) is one of seven EFE components from Eisenia fetida, and among them it has nearly the highest fibrinolytic activity. Here, we report its crystal structure at a resolution of 2.06A. The structural analysis shows that EFE-b should be classified as a trypsin from earthworm. However, it is distinct from other trypsins. It is a two-chained protease with an N-terminal, pyroglutamated light chain and an N-glycosylated heavy chain. Furthermore, the heavy chain contains a novel structural motif, an eight-membered ring resulting from a disulfide bridge between two neighboring cysteine residues, and a cis peptide bond exists between these two cysteine residues. The crystal structure of EFE-b provides the structural basis for its high level of stability and reveals its complicated post-translational modifications in earthworm. This structure is the first reported for a glycosylated two-chained trypsin, which may provide useful clues to explain the origin and evolution of the chymotrypsin family.

Amino Acid Sequence↗

PubMed2004

Influence of copper fungicide residues on occurrence of earthworms in avocado orchard soils.

The compost worm Eisenia fetida was used to demonstrate the avoidance by worms of Cu contaminated soil. Soils were collected from two avocado orchards in north eastern New South Wales, Australia. In avoidance trials, worms preferred non-contaminated control soils, sourced from adjacent to the orchard or an OECD control soil, when Cu residues in the orchard soils reached 4-34 mg Cu kg(-1). At levels of 553 mg Cu kg(-1), 90% avoidance of orchard soil was observed. The worms showed preference for the soils in the order; uncontaminated field derived soil >OECD standard soil >Cu contaminated orchard soil. It was demonstrated that OECD standard soil was less favoured by worms than control soil derived from the test sites. While Cu was found to be the primary influence on worm avoidance in orchard soil, other factors, such as elevated soil Zn concentrations, could not be discounted. In a corresponding field study, it was shown that earthworms occurred at lower density in orchard soils with a history of Cu fungicide use. In one such orchard, soil Cu concentrations of up to 270 mg kg(-1) were determined and no earthworms were found, while nearby control sites and less contaminated sites within the orchard had up to 40.7 g m(-2) earthworm biomass. Considering the potential for Cu to accumulate in these soils and the subsequent impacts on soil biota, our results highlight the importance of limiting future application of Cu based fungicides.

Agriculture↗

PubMed2005

Distribution of Cd, Zn, Cu and Fe among selected tissues of the earthworm (Allolobophora caliginosa) and Eurasian woodcock (Scolopax rusticola).

We have measured the concentrations of heavy metals in soils, earthworms and tissues of woodcocks in Quaderna Valley, northern Italy. The soil concentration of metals analysed in this research is consistent with data reported by other authors for uncontaminated or slightly contaminated soils. In earthworms, metals were mostly accumulated in the encapsulating chloragogenous tissue; the positive correlation between Cu concentration in the soil and in earthworms is noteworthy. Heavy metal distribution in the tissues of woodcock showed that Cd accumulation in the kidney was linked to the diet. Cu and Fe were preferentially concentrated in the liver and Zn in the testis. Kidney Cd and Zn concentrations were higher in adults than in juveniles. In addition, a main kidney metallothionein isoform, containing Cd and Zn, was isolated. In the kidney, Cd levels were linearly correlated with the concentration of metallothionein. Of the investigated metals, Cd raises the greatest concern, due to the increasing soil contamination by human activities.

Animals↗

PubMed2006

Lead concentrations in feathers and blood of common blackbirds (Turdus merula) and in earthworms inhabiting unpolluted and moderately polluted urban areas.

Despite the dramatic decrease of atmospheric lead (Pb) concentrations in urban areas of most industrialised countries, we hypothesised that urban common blackbirds (Turdus merula) may still be contaminated by Pb concentrations of toxicological concern due to transfer from soil through the food chain. We sampled blackbirds and earthworms, one of their main preys, in Besançon, a middle-size city of Eastern France (where atmospheric Pb concentrations decreased from 0.5 microg/m(3) in 1987 to nearly 0 in 2002) and in a rural reference site. Lead concentrations were determined in the tissues of the different functional groups of earthworms (anecic, epigeous and endogeous) and in blood, washed and unwashed outermost tail feathers and breast feathers of blackbirds. Fresh masses and an index of individual body condition were measured in the two blackbird populations as biomarkers of possible toxic effects. Lead concentrations in earthworms did not differ among functional groups but were significantly higher in urban individuals than in rural ones. Concentrations in outermost tail feathers, breast feathers and blood were significantly higher in urban blackbirds (7.75+/-4.50, 3.15+/-1.77 and 0.15+/-0.09 microg/g, respectively) than in rural individuals. In urban blackbirds, concentrations in washed and unwashed outermost tail feathers allowed estimating the external contamination (probably due to deposition of dusts and/or to excretion of the uropygial gland) at 37% of the total Pb concentration of the unwashed feathers. Remaining 63% should be linked to food chain transfer of persistent Pb from urban soils. Among the 23 sampled blackbirds, 4 of them (3 in the urban site and 1 in the rural site) exhibited blood Pb concentrations higher than the benchmark value (0.20 microg/g) related to subclinical and physiological effects in birds. Variations in body condition index were not correlated to Pb concentrations in blackbird tissues. Present results suggest that Pb may still be of environmental concern for blackbirds in urban areas because of the persistence of Pb in soils and its transfer through the food chain.

Animals↗

PubMed2003

Comparative toxicity of tetra ethyl lead and lead oxide to earthworms, Eisenia fetida (Savigny).

Leaded gasoline contains tetra ethyl lead (TEL) as an antiknocking agent, which produces major amounts of lead oxide in automobile exhaust along with traces of TEL. To minimize the lead contamination, methyl tertiary butyl ether (MTBE) is used as a substitute for producing unleaded gasoline. It has become increasingly apparent that young children are highly susceptible to the harmful effects of lead. Hence, a study was carried out to monitor lead toxicity in soil, using adult earthworms (Eisenia fetida, Savigny). Leaded gasoline (TEL) and lead oxide are 383- and 211-fold more toxic than unleaded gasoline (MTBE) in 7 days of exposure and 627- and 290-fold more toxic in 14 days, respectively. Results indicate that the presence of TEL in leaded gasoline and lead oxide has a significant effect on behavior, morphology, and histopathology of earthworms. Absorption of TEL into the tissues is comparatively less than that of lead oxide but toxic effects were severe. Rupture of the cuticle, extrusion of coelomic fluid and inflexible metameric segmentation were observed, causing desensitization of the posterior region leading to fragmentation in earthworms.

Animals↗

PubMed1998

Mixture toxicity and tissue interactions of Cd, Cu, Pb and Zn in earthworms (Oligochaeta) in laboratory and field soils: a critical evaluation of data.

Soil organisms inhabiting contaminated field sites are usually exposed to mixtures of toxicants. In such mixtures, toxicants can interact to express enhanced or weakened toxicity. Therefore, mixture effects should be considered in risk assessment methods for polluted soils. Data on mixture toxicity to soil organisms are scarce, however. In this paper, data on sublethal toxicity and tissue concentrations of Cd. Cu, Pb and Zn mixtures in earthworms are evaluated and compared with data on other organisms. Toxic effects were mainly antagonistic for total soil concentrations and nearly concentration-additive for 0.01 M CaCl2-extractable soil concentrations. Evidence to support concentration-additive behaviour of metals was found in interaction patterns within earthworm tissues. A method is proposed to assess the sublethal toxicity of metal mixtures in field soils to earthworms, based on single metal experiments from the laboratory. Finally, suggestions are made on how to incorporate mixture toxicity in risk assessment for polluted soils.

Animals↗

PubMedDate not supplied

Characterization of a 14 kDa plant-related serine protease inhibitor and regulation of cytotoxic activity in earthworm coelomic fluid.

We have purified and characterized the serine protease inhibitor activity contained in the coelomic fluid of the earthworms, Eisenia. Serine protease inhibitor activity was stable between pH3 and 9.5, not flocculable by pH 3.0 and resistant to 100 degrees C for 15 min. or to 4 degrees C for 24 h. Ten microL of coelomic fluid was sufficient to inhibit in vitro the protease activity of 0.12 microgram of trypsin. Injection of living bacteria into earthworms resulted in increased serine protease activity 1-2 days post-injection, and increased serine protease inhibitor activity on day 4, suggesting that serine protease inhibitor is responsible for serine protease neutralization. Purified to homogeneity by affinity chromatography on trypsin, the serine protease inhibitor of Eisenia is a monomer of 14 kDa. Its partial NH2 amino acid sequence revealed a basic hydrophobic fragment which shared 68-75% homologies and 47-60% identities with several plant serine protease inhibitors. Eisenia cytotoxic activity due to the two fetidins of 40 and 45 kDa was stimulable in vitro by several serine proteases. Incubation with soybean trypsin inhibitor variant a (STIa) resulted in less cytotoxicity. The inhibitory effect occurred only when STIa was added before cell disruption. Interpretative cytotoxic scheme involving the release of intracellular cytotoxic proteins, intracellular trypsin-like activator and extracellular serine protease inhibitor suggests regulatory mechanisms for cellular/humoral immune system of earthworms.

Aeromonas hydrophila↗

PubMedDate not supplied

Peptide fragments induce a more rapid immune response than intact proteins in earthworms.

The effect of in vivo proteolytic processing of protein antigen was studied in Eisenia foetida earthworms. Parenteral administration of the protein antigen induces elevated levels of an antigen-binding protein (ABP) which recognizes the protein used for stimulation. When the protein antigen is administered simultaneously with nontoxic serine proteinase inhibitor, ABP levels remain close to background. On the other hand, the in vivo adaptive response of earthworms to peptide fragments obtained by coelomic fluid digestion of the foreign antigen occurs even in the presence of proteinase inhibitor and, moreover, is significantly faster as compared to the response to intact antigen. These findings confirm the role of proteolytic processing in earthworms. MALDI mass spectrometric analysis of the fragments after coelomic fluid digestion has revealed the presence of the peptide fragments with molecular weights in the mass range 700-1100 Da.

Animals↗

PubMed2001

Visualization of nitric oxide production in the earthworm ventral nerve cord.

Distribution of nitric oxide (NO)-producible neurons in the ventral nerve cord (VNC) of the earthworm was investigated by nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry. Some neurons (20-30 microm in diameter) were intensely stained and were localized in areas between the 1st and 2nd lateral nerves in the ventral side of VNC. In contrast, no neurons including giant fibers were stained in the dorsal side. Endogenous NO production from VNC was visualized using a fluorescent dye, diaminofluorescein-2 diacethyl (DAF-2 DA). When VNC was incubated in a saline, a relative high level of NO was produced from the ventral side, especially from NADPH-d-positive neurons. Under high-K+ stimulation, NO was also detected in the giant fibers in the dorsal side of VNC. Our results suggest that the earthworm VNC constantly and relative highly produces NO as a neuromodulator, and that NO produced from the ventral side sometimes reaches and affects the giant fibers. In conclusion, we successfully visualized NO in the earthworm VNC by clarifying both the distribution of NO-producible neurons and the endogenous NO production.

Animals↗

PubMed2003

Interactions between earthworms and arsenic in the soil environment: a review.

Chemical pollution of the environment has become a major source of concern. In particular, many studies have investigated the impact of pollution on biota in the environment. Studies on metalliferous contaminated mine spoil wastes have shown that some soil organisms have the capability to become resistant to metal/metalloid toxicity. Earthworms are known to inhabit arsenic-rich metalliferous soils and, due to their intimate contact with the soil, in both the solid and aqueous phases, are likely to accumulate contaminants present in mine spoil. Earthworms that inhabit metalliferous contaminated soils must have developed mechanisms of resistance to the toxins found in these soils. The mechanisms of resistance are not fully understood; they may involve physiological adaptation (acclimation) or be genetic. This review discusses the relationships between earthworms and arsenic-rich mine spoil wastes, looking critically at resistance and possible mechanisms of resistance, in relation to soil edaphic factors and possible trophic transfer routes.

Adaptation, Physiological↗

PubMed1997

Cu accumulation in the earthworm Dendrobaena veneta in a heavy metal (Cu, Pb, Zn) contaminated site compared to Cu accumulation in laboratory experiments.

Earthworms (Dendrobaena veneta) were exposed to heavy metal contaminated soil from a field site under both laboratory and field conditions. In the laboratory study, D. veneta were analysed for Cu after 2 weeks' exposure. The tissue Cu concentration (Cu(W)) increased proportionally with the total extractable soil Cu content (Cu(T)) in soils that contained less than 150 mg Cu kg(-1). In D. veneta exposed to soils containing more than Cu(T) = 150 mg kg(-1), however, we observed no further increase in Cu(W). In a 64-m(2) field plot at a contaminated site, 81 soil samples were taken and analysed for Cu, Pb, Zn and pH. In June 1994, 1000 D. veneta were homogeneously distributed in the plot and sampled after 1, 2 and 5 weeks to analyse for Cu. Additionally, 2000 D. veneta were introduced in September 1994 and sampled after 1, 2 and 4 weeks. At locations where earthworms were removed, the soil Cu content was estimated by disjunctive kriging. Cu concentration factors (ratio of Cu(W) to Cu(T)) in the field experiment were in excellent agreement with those of the laboratory experiment. This indicates that, with a sufficiently high sampling density, it may be feasible to predict earthworm Cu accumulation in the field using a concentration factor determined in the laboratory.

Journal Article↗

PubMedDate not supplied

Earthworm muscle preparation for studying several potent compounds--the possible existence of a "Soman receptor.".

A new model of earthworm (Eisenia foedida) dorsal muscle preparation was studied. It is a specific preparation for testing GABA and its agonist muscimol. The response of muscle could be recorded through mechanical functional changes instead of potential variations. Thus, it is a direct and simple means for measuring GABA activity. ChE-deactivated preparation was suggested for this purpose. By mild membrane modifications of earthworm muscle preparation, it was possible to remove or inactivate GABA, glutamic acid or Soman combining protein. In conjunction with superfusion of Ach-receptor antagonist (alpha-Bungaratoxin) or GABA antagonist (picrotoxin, isopropylbicyclophosphate), we could differentiate Soman receptor from Ach and GABA receptor. The possible existence of a Soman receptor in the earthworm muscle was therefore proposed.

Animals↗

PubMed1999

Optical monitoring of the neural activity evoked by mechanical stimulation in the earthworm nervous system with a fluorescent dye, FM1-43.

In the central nervous system of the earthworm, sensory and motor neurons have direct synapses on three giant fibers. To determine the locations of synapses and neural network activated by mechanical stimuli, we optically monitored the activity-dependent staining in the earthworm ventral nerve cord with a styryl dye, N-(3-triethylammoniumpropyl)-4- (4-(dibutylamino)styryl)pyridinium dibromide (FM1-43), and a confocal laser scanning microscope. When scratch stimulus was applied to the body wall of the earthworm, bright fluorescent spots with 3-10 microns in diameter localized only in the stimulated segmental ganglion of the ventral nerve cord. The fluorescent intensity of these spots decreased during dye-free high K+ saline incubation. These results suggest that FM1-43 is useful for activity-dependent staining of invertebrate neurons and their synaptic regions as well as vertebrate nervous system.

Animals↗

PubMed1997

Molecular cloning of cDNA for lysenin, a novel protein in the earthworm Eisenia foetida that causes contraction of rat vascular smooth muscle.

Lysenin, which causes contraction of rat vascular smooth muscle, is a protein that was isolated from the earthworm Eisenia foetida. A cDNA encoding lysenin was isolated by use of a partial cDNA probe that had been generated by the PCR with a primer designed by reference to an internal peptide sequence of lysenin. This clone had an ORF encoding 297 amino acid residues. The amino acid sequence deduced from the cDNA revealed the absence of any significant homology to those of previously characterized vasoactive substances. The recombinant lysenin was produced in Escherichia coli. This protein and native lysenin isolated from the earthworm had similar contractive activities when tested on rat aorta. Northern blot analysis of the RNA from various tissues of the earthworm indicated that lysenin is produced by the coelomocytes.

Amino Acid Sequence↗

PubMed2001

[Diet of an anecic earthworm from the Colombian savannas: questioning ecological groups].

An ecological classification based on a set of morphological characters may be used to describe earthworm functions in soil. However these relationships have been seldom verified. Gizzards of the earthworm Martiodrilus carimaguensis (Glossoscolecidae) were studied to find out if its anecic morphology was in agreement with its diet (shallow litter remains and deep organo-mineral material). By direct observation of 13 adults' gizzard contents with a dissecting and a phase contrast microscope, the volumetric percentage of plant fragments, roots, amorphous organic matter and mineral particles reached respectively 63, 5, 6.5 and 30% v/v. Eighty percent of plant fragments were microscopic, and mostly incorporated into the organo-mineral earthworm casts well preserved in the gizzard. Coprophagy and rhizophagy were the dominant feature of M. carimaguensis diet. Yet, the casts produced revealed a high variability in its diet, what could be related to transitional adoption of a typical anecic diet during some periods.

Animals↗

PubMed2001

Potential of two epigeic and two anecic earthworm species in vermicomposting of water hyacinth.

The potential of two epigeic species (Eudrilus eugeniae Kinberg, and Perionyx excavatus Perrier) and two anecic species (Lampito mauritii Kinberg and Drawida willsi Michaelson) of earthworms was assessed in terms of efficiency and sustainability of vermicomposting water hyacinth (Eichhornia crassipes, Mart. Solm.). In different vermireactors, each run in duplicate with one of the four species of earthworms, and 75 g of 6:1 water hyacinth:cowdung as feed, vermicasts were produced with steadily increasing output in all the reactors. E. eugeniae was by far the most efficient producer of vermicasts, followed by the other epigeic P. excavatus. The two anecics came next, with D. willsi being the least effective which could generate only about half the quantity of vermicasts achieved in a corresponding time by E. eugeniae. In all the reactors, the earthworms grew well, increasing their weights by more than 250%. The maximum net gain of weight (average 30.7 g) was by E. eugeniae, followed by P. excavatus, L. mauritii and D. willsi. This trend, which followed the efficiency of vermicast production, was also shown in terms of reproductive ability as measured by the number of offspring produced by the four species.

Animals↗