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PubMed2002

Earthworm leukocytes react with different mammalian antigen-specific monoclonal antibodies.

We identified conserved molecules (enzymes, peptides, cytokines) that might play a role in invertebrate innate immunity. We found these molecules by immunoserological and immunohistochemical methods in association with coelomocytes, leukocytes located in the coelomic cavity of the earthworm Eisenia foetida. We detected the enzyme Cu-Zn-superoxide-dismutase (SOD), cytokines (tumor necrosis factor-alpha, TNFalpha; transforming growth factor-alpha, TGFalpha; and alpha peptide hormone, thyreotrope stimulating hormone, TSH) in earthworm coelomocytes with monoclonal antibodies developed originally against human and/or mouse antigens. Three coelomocyte subpopulations were identified according to their form, size and granularity by microscopic and flow cytometric analysis. These cell populations showed different reactivity with antibodies against mammalian cell surface (CD) markers and different intracellular antigens. Two coelomocyte types showed cell surface positivity with anti-Thy-1 (CD90), CD24 and TNF-alpha antibodies. Strong cytoplasmic reaction was shown with anti-TNF-alpha and anti-SOD mAbs and a weaker but unambiguous reaction with thyroid stimulating hormone (TSH) in two cell populations. The third population was negative for all of the monoclonal antibodies. Our flow cytometric results were confirmed by confocal microscopy both on the cell surfaces and intracellularly.

Journal Article↗
PubMed2002

19F-NMR and directly coupled 19F/1H-HPLC-NMR spectroscopic investigations of the metabolism of the model ecotoxin 3-trifluoromethylaniline in the earthworm species Eisenia veneta.

1. The metabolic fate of the model ecotoxin 3-trifluoromethylaniline (3-TFMA) in earthworm was studied by (19)F- and directly coupled (19)F/(1)H-HPLC-NMR spectroscopy. Earthworms of Eisenia veneta spp. were subjected to the ecotoxin during a filter papercontact toxicity test at exposure levels of 1000, 100, 10, 1 and 0.1 micro g cm(-2). A metabolic profile was obtained previously by (19)F-NMR spectroscopy and metabolites were observed at all the exposure levels. 2. Identification of metabolites in individual worm extracts at the (lethal) exposure levels of 1000 and 100 micro g cm(-2) could be achieved on-line without sample preparation by (19)F/(1)H-HPLC-NMR spectroscopy. (19)F-HPLC-NMR spectroscopy was used in the continuous-flow mode, which enabled the HPLC chromatographic retention times (t(R)) of the metabolites to be established in a single analytical step. 3. In total, three (19)F-NMR signals could be detected, of which one was identified as the parent compound. Two earlier eluting metabolites were identified to be alpha- and beta-glucoside conjugates of 3-TFMA. 4. Metabolites at the lower (sublethal) exposure levels of 10, 1 and 0.1 micro g cm(-2) escaped identification by (19)F/(1)H-HPLC-NMR spectroscopy as outlined here and will require concentration prior to analysis.

Aniline Compounds↗
PubMed2006

Expression and characterization of earthworm Eisenia foetida lumbrokinase-3 in Pichia pastoris.

Lumbrokinase-3 (LK-3, AY438622), first cloned from the earthworm Eisenia foetida in our laboratory, is a component of earthworm fibrinolytic enzymes. In this study, cDNA encoding the LK-3 gene was sub-cloned into yeast pPIC9K expression vector and transformed into the Pichia pastoris GS115 cells by electroporation. High level expression of LK-3 in yeast cells was confirmed with a different induction time. The activity of expressed LK-3 was observed in fibrin plates. In addition, the expressed LK-3 protein could dissolve fibrinogen and bovine serum albumin. The use of this system for the high level production of biological protein is implicated from this study.

Animals↗
PubMed2006

Influence of citric acid amendments on the availability of weathered PCBs to plant and earthworm species.

A series of small and large pot trials were conducted to assess the phytoextraction potential of several plant species for weathered polychlorinated biphenyls (PCBs) in soil (105 microg/g Arochlor 1268). In addition, the effect of citric acid on PCB bioavailability to both plants and earthworms was assessed. Under small pot conditions (one plant, 400 g soil), three cucurbits (Cucurbita pepo ssp pepo [zucchini] and ssp ovifera [nonzucchini summer squash], Cucumis sativus, cucumber) accumulated up to 270 microg PCB/g in the roots and 14 microg/g in the stems, resulting in 0.10% contaminant removal from soil. Periodic 1 mM subsurface amendments of citric acid increased the stem and leaf PCB concentration by 330 and 600%, respectively, and resulted in up to a 65% increase in the total amount of contaminant removed from soil. Although citric acid at 10 mM more than doubled the amount of PCB desorbed in abiotic batch slurries, contaminant accumulation by two earthworm species (Eisenia foetida and Lumbricus terrestris) was unaffected by citric acid at 1 and 10 mM and ranged from 11-15 microg/g. Two large pot trials were conducted in which cucurbits (C. pepo ssp pepo and ssp ovifera, C. sativus) and white lupin (Lupinus albus) were grown in 70 kg of PCB-contaminated soil White lupin was the poorest accumulator of PCBs, with approximately 20 microg/g in the roots and 1 microg/g in the stems. Both C. pepo ssp ovifera (summer squash) and C. sativus (cucumber) accumulated approximately 65-100 microg/g in the roots and 6-10 microg/g in the stems. C. pepo ssp pepo (zucchini) accumulated significantly greater levels of PCB than all other species, with 430 microg/g in the roots and 22 microg/g in the stems. The mechanism by which C. pepo spp pepo extracts and translocates weathered PCBs is unknown, but confirms earlier findings on the phytoextraction of other weathered persistent organic pollutants such as chlordane, p,p'-DDE, and polycyclic aromatic hydrocarbons.

Animals↗
PubMed2003

Nutrient content of earthworms consumed by Ye'Kuana Amerindians of the Alto Orinoco of Venezuela.

For the Makiritare (Ye'Kuana) native people of the Alto Orinoco (Venezuela), earthworms (Anellida: Glossoscolecidae) are an important component of the diet. Two species in particular are widely consumed: 'kuru' (Andiorrhinus kuru n. sp.) and 'motto' (Andiorrhinus motto). We analysed eviscerated kuru body proper, and whole and smoked preparations of motto for their content of protein and amino acids, fatty acids and 20 minerals and trace elements. The samples contained large amounts of protein (64.5-72.9% of dry weight), essential amino acids, calcium and iron together with notable quantities of other important elements, indicating that these earthworms contain potentially useful quantities of many nutrients that are critical to the health of the humans who consume them.

Amino Acids↗
PubMed2000

Crystallization and preliminary X-ray analysis of earthworm fibrinolytic enzyme component A from Eisenia fetida.

Earthworm fibrinolytic enzyme component A, a protein which functions both as a direct fibrinolytic enzyme and a plasminogen activator, was purified from the earthworm Eisenia fetida. Diffraction-quality single crystals of the protein were grown by the hanging-drop vapour-diffusion technique with ammonium sulfate as a precipitant. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 40.6, b = 127.5, c = 129.2 A and three molecules per asymmetric unit. The data set reached a resolution of 1.95 A.

Animals↗
PubMed1985

Lipid composition of the nervous system of earthworms (Lumbricus terrestris).

As part of a systematic study of the evolution of the nervous system, the lipid composition of the ventral nerves of earthworms was examined. The nerve axons are wrapped in copious layers of loosely bound membrane, superficially resembling the myelin sheath of vertebrates. However, neither galactocerebroside nor sulfatide, both of which are considered to be markers for myelin, was present, and only traces of glucocerebroside, which is abundant in shrimp nerve, were detected. The lipids were rich in cholesterol (15.3 mumol/g of fresh tissue) and phospholipids (21.7 mumol/g of fresh tissue). The phospholipids were composed of phosphatidylethanolamine, -choline, -serine, and -inositol in the ratio of 41:44:8:7. Most of the ethanolamine-containing phospholipids were in the form of plasmalogens. The fatty acid moieties of these phospholipids were predominantly 18:1, 18:0, and 20:1, whereas the aldehyde moieties of plasmalogen were mostly 18:0. Sphingomyelin, which is considered a ubiquitous component of animal membranes, was not detected. How the unique structure of the membranes of earthworm nerves may be related to the function of the nervous system in this organism is discussed.

Animals↗
PubMed1963

CARBAMYL PHOSPHATE SYNTHESIS IN THE EARTHWORM LUMBRICUS TERRESTRIS.

The enzymatic synthesis of citrulline from ammonia, bicarbonate, adenosine triphosphate, and L-ornithine takes place in the soluble fraction of gut tissue of the earthworm. The synthesis occurs at low ammonia concentrations, is dependent upon added N-acetyl-L-glutamate, and results in incorporation of the bicarbonate carbon into the ureido group of the citrulline molecule. Carbamyl phosphate is the intermediate in the reaction and its biosynthesis in the earthworm is mediated by a carbamyl phosphate synthetase system similar to that of ureotelic vertebrates.

Adenosine Triphosphate↗
PubMed1968

Alarm pheromone in the earthworm Lumbricus terrestris.

Noxious stimulation of the earthworm Lumbricus terrestris elicits secretion of a mucus that is aversive to other members of the species, as well as to the stimulated animal when it is encountered later. This alarm pheromone is not readily soluble in water and retains its aversive properties for at least several months if not disturbed. Its influence may be responsible for some features of the data on instrumental learning in earthworms.

Animals↗
PubMedDate not supplied

Endogenous lipids of the earthworm Lumbricus terrestris.

Earthworms (Lumbricus terrestris) were given [1-14C]-labeled palmitic acid by gavage on days 0 and 3, and sacrificed on day 7. The distribution of label among lipid classes indicated that glycerides, sterol esters, cerebrosides, sulfatides, phosphatidylethanolamine, phosphatidylserine and (or) phosphatidylinositol, phosphatidylcholine, and sphingomyelin turn over in, or are synthesized by, the earthworm. Free fatty acids still had the highest specific radioactivity of any lipid class at the end of the experiment. Incorporation of label into sterol and hydrocarbon fractions was insignificant and there was no detectable label incorporated into gangliosides. Phosphatidylethanolamine apparently turned over quite slowly compared with other lipid classes, while the cerebroside fraction became highly labeled. Elongation of palmitic acid to stearate and oxidation to CO2 occurred extensively, but there was no evidence for desaturation.

Animals↗
PubMed1991

A non-invasive technique for sequential collection of earthworm (Lumbricus terrestris) leukocytes during subchronic immunotoxicity studies.

We have compared two methods for collecting earthworm leukocytes (coelomocytes) with respect to cell yield, viability and behaviour in immunoassays. Non-invasive extrusion was more efficient than puncturing the coelomic cavity. Extrusion does not produce trauma to earthworms maintained under long term laboratory conditions. Neither technique modified immune assays, as determined by erythrocyte and secretory rosette formation, and phagocytosis, since all 3 were functionally equivalent. After an initial extrusion, sequential leukocyte collections by extrusion are possible at intervals of 6 weeks without affecting total and differential cell counts and rosette formation.

Animals↗
PubMed2001

Molecular cloning, sequencing, and expression of cDNA encoding serine protease with fibrinolytic activity from earthworm.

An earthworm, Lumbricus rubellus, produces alkaline trypsin-like proteases that are greater than trypsins in their stability and strong tolerance to organic solvents. cDNAs encoding strong fibrinolytic proteases (F-III-2 and F-III-1) in the six isozymes were cloned and sequenced to study their stability-structure relationship. The cDNAs of F-III-2 and F-III-1 comprised 1011 and 973 bp and included open reading frames that encode polypeptides of 245 and 246 amino acid residues, respectively. The deduced amino acid sequences of F-III-2 and F-III-1 have 7 and 8 activation peptides in the N-termini respectively, indicating that they are translated as proenzymes and modified to active forms by posttranslational processing. They showed similarity to mammalian serine proteases and conserved the catalytic amino acid residues, however, neither arginine nor lysine residues were present in the autolysis region. The gene encoding the native form of F-III-2 was expressed in Pichia pastoris to produce and secrete the earthworm protease in the culture medium, which dissolves an artificial fibrin plate.

Amino Acid Sequence↗
PubMed2006

Visceral larva migrans associated with earthworm ingestion: clinical evolution in an adolescent patient.

A 16-year-old girl developed a cough, hypereosinophilia (absolute eosinophil count: 32000/mm3), hypergammaglobulinemia, and multiple noncavitary pulmonary nodules 1 month after having ingested an earthworm on a dare. Spirometry revealed moderate restriction and reduced gas diffusion. Parabronchial biopsy demonstrated eosinophilic organizing pneumonitis with multiple eosinophilic microabscesses, and Toxocara titers were elevated (>1:4096). Ophthalmologic examination ruled out ocular larva migrans. The patient received a 10-day course of albendazole (400 mg orally twice daily) and demonstrated significant clinical improvement with resolution of cough and pulmonary function abnormalities. Her white blood cell count and hypergammaglobulinemia normalized within 20 days, yet eosinophils (absolute eosinophil count: 1780/mm3) and Toxocara serologies (>1:4096) remained elevated 3 months after completing antihelminthic therapy. In this instance, the ingested earthworm served as the paratenic carrier of Toxocara larvae from the soil to the patient. This case highlights the clinical evolution of pulmonary visceral larva migrans infection caused by Toxocara spp. associated with a discrete ingestion in an adolescent patient. In addition, it provides a rare opportunity to define the incubation period of visceral larva migrans and emphasizes the importance of education regarding sources of Toxocara infection.

Adolescent↗
PubMed2005

Effect of volatile hydrocarbon fractions on mobility and earthworm uptake of polycyclic aromatic hydrocarbons from soils and soil/lampblack mixtures.

Studies were conducted to examine the mobility and bioavailability to earthworms (Eisenia fetida) of priority pollutant polycyclic aromatic hydrocarbons (PAH) in a suite of 11 soils and soil/lampblack mixtures obtained from former manufactured-gas plant sites. Contaminant mobility was assessed using XAD4 resins encapsulated in dialysis tubing, which were exposed to slurried soils for 15 d. These experiments showed that mobility of PAH in the different soils strongly correlated to the levels of volatile hydrocarbons (namely, gasoline- and diesel-range organics [GRO and DRO]) that existed in the soils as co-contaminants. Actual PAH bioavailability (as measured by earthworm PAH concentrations) also appeared to depend on GRO + DRO levels, although this was most evident at high levels of these contaminants. These findings are discussed in view of the effects of dieselrange organics on oil viscosity, assuming that the hydrocarbon contaminants in these soils exist in the form of distinct adsorbed oil phases. This study, therefore, extends correlations between carrier-oil viscosity and dissolved solute bioavailability, previously observed in a number of other in vitro and whole-organism tests (and in bacterial mutagenicity studies in soil), to multicellular organisms inhabiting contaminated-soil systems.

Animals↗
PubMed2006

Tungsten effects on survival, growth, and reproduction in the earthworm, Eisenia fetida.

To provide basic toxicity data for formulating risk characterization benchmarks, the effects of tungsten on survival, growth, and reproduction were investigated in the earthworm Eisenia fetida. Parallel studies with lead as a reference toxicant also were conducted. Although sodium tungstate (Na2WO4) was less acutely toxic than lead nitrate (Pb(NO3)2) in 14-d spiked field soil acute toxicity assays (lethal concentrations for 50% of organisms: W, 6,250 mg/kg; Pb, 2,490 mg/kg), tungstate completely inhibited reproduction in 28- and 56-d assays at all tested tungsten concentrations (> or = 704 mg/kg). By comparison, cocoon production was not significantly reduced for lead concentrations until concentrations reached 766 mg/kg, and cocoon production was still observed at the highest concentration tested (1,650 mg/kg). These data indicate that tungsten is a reproductive toxicant for earthworms and that, by comparison, its sublethal toxicity is greater than that of lead. Toxicity data for other soil invertebrate species are required to fully establish benchmark levels/ecological soil screening levels for tungsten.

Animals↗
PubMed2001

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 4. The influence of salt speciation, soil type, and sewage sludge on toxicity using the collembolan Folsomia fimetaria and the earthworm Aporrectodea caliginosa as test organisms.

Sewage sludge applied to agricultural soils often contains linear alkylbenzene sulfonates (LAS) in the range of 1 to 10 g/kg dry weight, and their toxicity to relevant soil organisms should, therefore, be assessed to ensure safe use of sewage sludge as a fertilizer. Studies of LAS toxicity to soil organisms are few, and to our knowledge, factors that may influence the toxicity in the field have not been studied in detail. In this paper, we report on the influence of speciation of LAS in the test solution added to soil (soluble Na-LAS vs poorly soluble Ca-LAS or Mg-LAS), the influence of soil type, and the modifying effects of sludge amendment on the toxicity of LAS. These issues were investigated using reproduction of Collembola and growth of juvenile earthworms as test parameters. Speciation of the LAS added to test soil did not have any influence on toxicity for any of the test species. Likewise, in three different agricultural soils (sand, loam, and clay), we found almost equal toxicities. The LAS added to test soil in a sludge-water suspension was equally toxic as when it was added in an aqueous solution. However, anaerobic incubation for 7 and 14 d of the LAS-sludge suspension (with no decay of LAS) caused the toxicity to increase almost threefold in both collembolan and earthworm. The relationships between soil constituents, bioavailability, and toxicity are also discussed.

Agriculture↗
PubMed2001

Resistance to copper toxicity in populations of the earthworms Lumbricus rubellus and Dendrodrilus rubidus from contaminated mine wastes.

Two arsenic and heavy metal-contaminated mine spoil sites, at Carrock Fell, Cumbria, United Kingdom, and Devon Great Consols Mine, Devon, United Kingdom, have been found to support populations of the earthworms Lumbricus rubellus Hoffmeister and Dendrodrilus rubidus (Savigny). Lumbricus rubellus and D. rubidus collected from the Devon site and an uncontaminated site were kept for 28 d in uncontaminated soil and in soil containing 750 mg/kg CuCl2, the state of the specimens being recorded using a semiquantitative assessment of earthworm health (condition index). The condition index remained high for all specimens except those of L. rubellus and D. rubidus from uncontaminated sites, which displayed 100% mortality. Bioavailability of Cu in the soils from one uncontaminated and two contaminated sites and in the uncontaminated soil treated with CuCl2 was determined using sequential extraction. Soils from Devon Great Consols had the greatest availability of Cu, Carrock Fell the lowest. Total tissue Cu for L. rubellus and D. rubidus from the contaminated sites did not change significantly for each species during the experiment. Total tissue concentrations of Cu for L. rubellus and D. rubidus from uncontaminated sites increased significantly during the first 7 d, after which mortality was 90%, making it impossible to continue the analysis.

Animals↗
PubMed2002

Morphological plasticity in metal-sequestering earthworm chloragocytes: morphometric electron microscopy provides a biomarker. of exposure in field populations.

Morphometric measurements of the volume fractions (Vv) of two prominent metal-sequestering compartments, chloragosome granules and debris vesicles, were made on transmission electron micrographs of chloragocytes in Dendrodrilus rubidus (Annelida: Oligochaeta: Lumbricidae) from one uncontaminated site and three qualitatively and quantitatively different metalliferous soils associated with disused Pb/Zn mines. We also examined the relative volume fractions of the alimentary wall and attached chloragogenous tissue by light microscopy. Several conclusions relevant to environmental diagnostics were reached: Metal identity and degree of soil pollution exert strong effects on chloragocyte ultrastructure; morphometry by light microscopy indicated that the metal-induced structural effects are characterized by intracellular alterations not accompanied by a significant expansion of the tissue volume; elevated metal burdens can reduce chloragosome Vv and, concomitantly, increase the Vv of their autophagic derivatives, the debris vesicles; the measured cellular alterations were more closely correlated with integrated tissue metal burdens than they were with integrated soil metal burdens; and estimates of tissue metal distribution, derived by combining morphometric data and published whole-worm and chloragosome metal concentrations, suggested that the alimentary wall of earthworms may sequester Pb, Zn, and Cd. Our study shows that cellular changes, directly or indirectly, may yield quantifiable expressions of metal loads bioaccumulated by earthworms. However, the practical use of cellular changes as biomarkers of soil quality probably will require automated light-microscopic morphometry in combination with organelle-specific molecular probes to be cost effective.

Animals↗