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[Histopathologic study of the host-parasite relationship: the earthworm-, wild boar-metastrongyle model].

A histopathological study was conducted on two hosts of metastrongyles (Metastrongylus sp.): earthworms and wild boars (Sus scrofa Linnaeus, 1758). In the earthworm intermediate host there were rare granulomas outside the normal site occupied by larvae (blood sinuses of calciferous glands). Given the small number of these reactions, they could not constitute a limiting factor for parasitism in this host. However, the lungs of wild boars contained numerous lesions associated with the presence of these nematodes. Such lesions, often inflammatory, were part of an immunological defence mechanism which controls the extent of parasitism in wild boar.

Animals↗

Assessment of metal availability in smelter soil using earthworms and chemical extractions.

Chemical immobilization is a relatively inexpensive in situ remediation method that reduces soil contaminant solubility, but the ability of this remediation treatment to reduce heavy metal bioavailability and ecotoxicity to soil invertebrates has not been evaluated. Our objectives were to (i) assess the ability of chemical immobilization amendments (municipal sewage sludge biosolids and rock phosphate) to reduce metal bioavailability and toxicity in a toxic metal-contaminated smelter soil and (ii) evaluate soil extraction methods using Ca(NO3)2 solution or ion-exchange membranes coated with diethylenetriaminepentaacetic acid (DTPA) as surrogate measures of metal bioavailability and ecotoxicity. We treated a soil contaminated by Zn and Pb milling and smelting operations and an uncontaminated control soil with lime-stabilized municipal biosolids (LSB), rock phosphate (RP), or anaerobically digested municipal biosolids (SS) and evaluated lethality of the remediated soils to earthworm (Eisenia fetida Savigny). Lime-stabilized municipal biosolids was the only remediation amendment to successfully immobilize lethal levels of Zn in the smelter soil (14-d cumulative mortality < or = 15%). Calcium nitrate-extractable Zn in the lethal Zn smelter soil-amendment combinations was 11.5 to 18.2 mmol/kg, compared with the nonlethal LSB amended soil (0.62 mmol/kg). The Ca(NO3)2-extractable Zn-based median lethal concentration (LC50) of 6.33 mmol/kg previously developed in Zn-spiked artificial soils was applicable in the remediated smelter soils despite a 14-fold difference in total Zn concentration. Chelating ion-exchange membrane uptake among the soils was highly variable (mean CV = 39%) compared with the Ca(NO3)2-extraction (mean CV = 1.9%) and not well related to earthworm toxicity.

Animals↗

A novel fibrinolytic enzyme extracted from the earthworm, Lumbricus rubellus.

A strong fibrinolytic enzyme was readily obtained in saline extracts of the earthworm, Lumbricus rubellus. It hydrolyzed not only plasminogen-rich fibrin plates, but also plasminogen-free fibrin plates. The average fibrinolytic activity was about 100 CU (plasmin units) or 250 IU (urokinase units)/g wet weight. The molecular weight and isoelectric point were about 20,000 and 3.4, respectively. The enzyme was heat-stable and displayed a very broad optimal pH range. DFP and SBTI strongly inhibited the enzyme, but the anti-plasmin agent, t-AMCHA, exerted little effect under the same conditions. Purification of the enzyme was performed and three partially purified fractions were obtained. These three fractions were further subdivided. The first fraction (F-I) was divided into three fractions (F-I-0, F-I-1, and F-I-2), which exhibited similar biochemical characteristics. The second fraction (F-II) could not be subdivided. The third fraction (F-III) was divided into two fractions (F-III-1 and F-III-2). Based on results for their enzymatic activities against various substrates, the fraction I enzymes are thought to represent a chymotrypsin-like enzyme and the fraction III enzymes to represent a trypsin-like enzyme. The fraction II enzyme appears to be neither a trypsin- or chymotrypsin-like enzyme nor an elastase. The amino acid compositions of the six enzymes were estimated. Compared with other serine enzymes, these enzymes contained very abundant asparagine or aspartic acid, and there was very little proline or lysine. From the above data, these enzymes are regarded as novel fibrinolytic enzymes, and we name them collectively as Lumbrokinase from the generic name of the earthworm.

Amino Acids↗

Isolation and some characterizations of a glycosylated fibrinolytic enzyme of earthworm, Eisenia fetida.

Resin coupled with m-aminophenylbornic acid was used to isolate a glycosylated component from homogenate of earthworm (Eisenia fetida). The fraction showed a single band on SDS-PAGE with a molecular weight of 34, 193 Da determined by mass spectroscopy. The N-terminal region is AQVCCPDI, different from those of earthworm fibrinolytic enzymes reported previously (Nakajima et al. 1993). This glycosylated component showed an activity on digesting both Chromozym-TH and fibrin, suggesting that it is a novel fibrinolytic enzyme.

Animals↗

Rapid induction of functional and morphological continuity between severed ends of mammalian or earthworm myelinated axons.

The inability to rapidly restore the loss of function that results from severance (cutting or crushing) of PNS and CNS axons is a severe clinical problem. As a novel strategy to help alleviate this problem, we have developed in vitro procedures using Ca2+-free solutions of polyethylene glycol (PEG solutions), which within minutes induce functional and morphological continuity (PEG-induced fusion) between the cut or crushed ends of myelinated sciatic or spinal axons in rats. Using a PEG-based hydrogel that binds to connective tissue to provide mechanical strength at the lesion site and is nontoxic to nerve tissues in earthworms and mammals, we have also developed in vivo procedures that permanently maintain earthworm myelinated medial giant axons whose functional and morphological integrity has been restored by PEG-induced fusion after axonal severance. In all these in vitro or in vivo procedures, the success of PEG-induced fusion of sciatic or spinal axons and myelinated medial giant axons is measured by the restored conduction of action potentials through the lesion site, the presence of intact axonal profiles in electron micrographs taken at the lesion site, and/or the intra-axonal diffusion of fluorescent dyes across the lesion site. These and other data suggest that the application of polymeric fusiogens (such as our PEG solutions), possibly combined with a tissue adherent (such as our PEG hydrogels), could lead to in vivo treatments that rapidly and permanently repair cut or crushed axons in the PNS and CNS of adult mammals, including humans.

Animals↗

[Effect of the medium composition on the resting membrane potential in the earthworm longitudinal somatic muscle fibers].

Norepinephrine or increased extracellular K+ hyperpolarize the membrane of the earthworm somatic muscle fibre, whereas removal of Cl- from external solution or a hypotonic solution depolarize the membrane. The dependence of the membrane resting potential on the extracellular K+ is quite characteristic against the background of ouabain action. A preliminary membrane depolarisation by ouabain eliminates the above effects on the membrane resting potential. The data obtained suggest that the ouabain-sensitive active ion pump directly contributes to the membrane resting potential value. This hypothesis is discussed with respect to existence of active Cl- transport combined with Na+, K(+)-pump which presumably takes part in the intracellular osmotic pressure regulation in the earthworm somatic muscle.

Animals↗

Neutral red retention by earthworm coelomocytes: a biomarker of cadmium contamination in soil.

The earthworm Metaphire posthuma were used as a model to assess the toxic potential of cadmium incorporated into the soil by environmental or human activities. The retention period of neutral red in the lysosomes of the coelomocytes was used as a biomarker. The viability of harvested coelomocytes by a non-invasive extrusion protocol was 93% with no alteration by the dye during experimentation. The control cells retained dye for 119 and 121 min in normal soil and KCl, respectively, whereas a linear decline in the retention time in the treated earthworm coelomocytes was observed. This illustrated that the presence of cadmium caused damage to the lysosomes of the coelomocytes.

Animals↗

[Dynamics of the biological activity of litter in course of its transformation by earthworms].

The dynamics of the biological activity of litter during its transformation were studied under laboratory conditions. The model group of soil saprophages was represented by earthworms (Eisenia genus). The system "litter-droppings-litter + droppings-soil" was investigated. The feeding activity of earthworms intensified biological processes in the decomposing plant residues. A fraction of readily hydrolyzable nitrogen was accumulated in the worm droppings. Dressing the litter with droppings shifted transformation of the plant materials towards mineralization.

Animals↗

Biomanagement of paper mill sludge using an indegenous (Lampito mauritii) and two exotic (Eudrilus eugineae and Eisenia foetida) earthworms.

Paper mills have severe problem in disposing effluent or semisolid sludge despite repeated recycling. It requires treatment prior to disposal of sludge. In recent years biological treatment methods received much attention and considered as efficient low-cost treatment. One such method is vermiculture treatment. The present study was carried out to dispose the paper mill sludge biologically using 2 exotic species (Eudrilus eugineae and Eiseniafoetida) and an indigenous species (Lampito mauritii) of earthworm. The paper mill sludge in various concentration 25%,50% and 75% were subjected to vermitub treatment for a period of 60 days. During the period of study data were collected on reproductive strategies of earthworm and chemical analysis of wastes before and after treatment. Results obtained indicate that 25% concentration of sludge was ideal and of the three worms used Eiseniafoetida proved to be the best worm for biomanagement.

Animals↗

Application of sewage sludge to arable land-soil concentrations of polybrominated diphenyl ethers and polychorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls, and their accumulation in earthworms.

Soils from five agricultural sites, three research sites, and two privately owned farms were analyzed for polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), and polybronimated diphenylethers (PBDEs). In soils that had not been treated with sludge (reference soils), the international toxic equivalents (I-TEQs) were 0.6 to 1.5 pg/g dry matter (D.M) for the PCDD/Fs. which are low compared to generally reported background soil levels in Europe. The concentrations of sum of six penta- and hexa-PCBs were 450 to 1,400 pg/g DM. The PBDEs 47 and 99 dominated among the tri- to hepta-PBDEs analyzed (8-80 pg/g DM). The impact of adding 1 to 3 tonnes of sludge (DM) per hectare per year on the concentrations was studied at the three research sites by calculating ratios of the compounds in sludge-treated soil to reference soil (S/R ratio). The concentrations of I-TEQs did not increase in the sludge-treated soil, whereas the S/R ratios for PBDEs were greater than one. Also, although the PCB contents were higher in the sludge-treated soils, the background sources were more important for the concentrations of PCBs than of PBDEs. The largest increase in the S/R ratios was found at a private farm where large amounts of sludge had been used in the past. Accumulation of the compounds in earthworms from the sites also was investigated. The biota-soil accumulation factors (BSAFs) of the tested compounds declined in the following order: ortho-PCBs approximately PBDEs > non-ortho-PCBs > 2.3,7,8-substituted PCDD/Fs. The average BSAF for ortho-PCBs was five (organic matter/lipids), and the lowest BSAFs (0.1-0.8) found were for octachlorodibenzo-p-dioxin. To our knowledge, accumulation of PBDEs in earthworms has not been published previously.

Agriculture↗

[Influence of different ecological groups of earthworms on the intensity of nitrogen fixation].

The results of studying nitrogen fixation in organic substrates processed by different ecological groups of earthworms suggest that the earthworms Aporrectoidea caliginosa actively stimulate nonsymbiotic nitrogen fixation. In this respect, they exceed the manure worms Eizenia foetida tens of times due to the formation in the organic substrate of conditions favorable for the nitrogen fixing bacteria, and namely: low content of nitrogen easily accessible for microorganisms, changes in the structure of the microbial community of the substrate in favor of non-spore forms of bacteria, and suppression of the growth of saprophyte bacilli, the main competitors of nitrogen fixing bacteria for carbon nourishment sources.

Animals↗

Biological effects of two insecticides on earthworms (Lumbricus terrestris L.) under laboratory conditions.

Earthworms (Lumbricus terrestris L.) were exposed to endosulfan and aldicarb for 2, 7, and 15 days and lethal concentrations were determined. Worms were then exposed to these concentrations (LC10, LC25, and LC50) of endosulfan and (LC10 and LC25) of aldicarb. Growth rate, total protein content, and insecticide residues were determined. Aldicarb was more toxic than endosulfan and both insecticides caused a significant reduction of growth rate and total protein content of earthworms. Residues remaining in the soil after 2 to 15 days ranged between 37.75% and 68.54% of applied concentration for endosulfan and between 10.13% and 67.71% of applied concentration for aldicarb. Less than 1% of soil concentration was detected in worms and accumulation was more important in the case of endosulfan. This study proposes the use of growth rate and total protein content as biomarkers for contamination by endosulfan and aldicarb.

Aldicarb↗

Arsenic speciation in the earthworms Lumbricus rubellus and Dendrodrilus rubidus.

Two species of earthworm, Lumbricus rubellus Hoffmeister and Dendrodrilus rubidus (Savigny) collected from an arsenic-contaminated mine spoil site and an uncontaminated site were investigated for total tissue arsenic concentrations and for arsenic compounds by liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS). For L. rubellus, whole-body total tissue arsenic concentrations were 7.0 to 17.0 mg arsenic/ kg dry weight in uncontaminated soil and 162 to 566 mg arsenic/kg dry weight in contaminated soil. For D. rubidus, whole-body tissue concentrations were 2.0 to 5.0 mg arsenic/kg dry weight and 97 to 321 mg arsenic/kg dry weight, respectively. Arsenobetaine was the only organic arsenic species detected in both species of earthworms, with the remainder of the extractable arsenic being arsenate and arsenite. There was an increase in the proportion of arsenic present as arsenobetaine in the total arsenic burden. Lumbricus rubellus and D. rubidus have similar life styles, both being surface living and litter feeding. Arsenic speciation was found to be similar in both species for both uncontaminated and contaminated sites, with dose-dependent formation of arsenobetaine. When L. rubellus and D. rabidus from contaminated sites were incubated in arsenic-free soils, the total tissue burden of arsenic diminished. Initially, L. rubellus from the tolerant populations (from the contaminated site) eliminated arsenic in the first 7 d of exposure before accumulating arsenic in tissues, whereas nontolerant populations (from the uncontaminated site) accumulated arsenic linearly. The tolerant and nontolerant L. rubellus eliminated tissue arsenic linearly over 21 d when incubated in uncontaminated soil.

Animals↗

Chamber and gavage technique for metabolic studies of earthworms.

Earthworms make very suitable laboratory animals for metabolic studies in vivo using radiolabeled test chemicals. We describe the construction and operation of a metabolic chamber to enable the collection of labeled CO2, volatile organics, material excreted into the bedding, and labeled material remaining in the worms. A gavage technique has been developed that permits the administration of water-soluble and lipid-soluble test chemicals in spite of the extremely low level of triglyceride lipase activity in the earthworm gut. This technique is less likely to puncture the worm tissue than previous methods. Radiolabeled DDT and diethylhexyl adipate were used to provide examples of the use of these techniques and the metabolic chamber. Results were qualitatively similar to those that have been noted in vertebrates.

Adipates↗

Chloride cotransport in the membrane of earthworm body wall muscles.

The resting membrane potential (V(m)) of isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris was studied by glass microelectrodes. The inhibition of chloride permeability by low pH did not affect V(m) of the muscle fibers in isolated somatic longitudinal muscles of the earthworm Lumbricus terrestris which was -48.7 mV (inside negative) at pH 7.3 and -49.1 at pH 5.6. On the other hand, bathing the muscles in Cl(-) and Na(+)-free solutions, or application of the chloride transporter inhibitor furosemide and Na(+)-K(+)-ATPase inhibitor ouabain depolarized the V(m) by 3-5 mV. The effects of a Cl(-) -free solution and ouabain were not additive. This demonstrates relatively small contribution of equilibrium potential for Cl(-) to the resting membrane potential and electrogenic effect of Na(+)K(+)-ATPase which is dependent on the supply of Na(+)(i) ions by furosemide-sensitive and Cl(-)(e)- and Na(+)(e)-dependent electroneutral transport (most probably Na(+)K(+)Cl(-) cotransport).

Animals↗

Seasonal changes in population of some selected earthworm species and soil nutrients in cultivated agroecosystem.

Studies were conducted on population dynamics of Metaphire posthuma, Lampito mauritii and Dichogaster bolaui in cultivated pedoecosystem of desert region of Rajasthan. The populations of aclitellate and clitellate M. posthuma, L. mauritii and D. bolaui were maximum in rainy season and minimum in summer season. The abundant population of these worms were found during the months of July to October. The species M. posthuma breed throughout the year except in extreme summer but L. mauritii breed twice in a year in the field. However, D. bolaui breed once in a year. Among these earthworm species D. bolaui with relative density of 44.2% was the most dominant species in cultivated land while L. mauritii and M. posthuma contributed 33.3% and 22.5% of density, respectively. The total population of earthworms showed significant positive correlation with different soil nutrients.

Agriculture↗

Biomanagement of petrochemical sludge using an exotic earthworm Eudrilus eugineae.

Petrochemical industry have severe problem in disposing effluent and semisolid sludge despite repeated recycling. It requires further treatment prior to disposal of sludge. In recent years biological treatment methods received much attention and considered as an efficient low-cost treatment. One such method is vermiculture treatment The end product of vermicompost is rich in essential micro and macronutrients along with microorganisms in a very simple form. Adding cast, not only improves the soil structure and fertility but also leads to improvement in overall plant growth and thus increase their yield. The present study was carried out to dispose the petrochemical sludge biologically using an exotic earthworm Eudrilus eugineae. The petrochemical sludge at various concentrations 25, 50 and 75% were subjected to vermicomposting treatment for a period of 60 days. During the period of study, data were collected on life form of earthworm and chemical analysis of the sludge before and after treatment. The microbial analysis was carried out fortnightly. The results indicate that 25 and 50% concentration of sludge was ideal for the vermicomposting, whereas the higher concentration inhibits the vermicomposting.

Animals↗

[Aging of spiked pyrene in two paddy soils and their particle-size fractions after soil incubation and changes in extractability and bio-availability to earthworm].

Effect of aging on bioavailability and extractability of persistent organic pollutants has recently been paid much attention in environmental studies. This study deals with aging of spiked pyrene, a four-ring polycyclic hydrocarbon, in two paddy soils and their particle-size fractions under laboratory incubation and change in its extractability and bioavailability. The bioavailability of aged pyrene was tested by an assay of exposure to earthworms (Eisenia foetida). The extractability of spiked pyrene was decreased by 15% to 23%, and the bioavailability to earthworm was decreased by 37% to 67% after incubation for 90 d compared to under no incubation. Meanwhile, there was no significant difference in the pyrene extractability between incubation with different size fractions of a single soil. Soil incubation resulted in lower extractability and, thus, lower environmental risk of the organic pollutants after incorporation in paddy soils and their aggregates. The different size fractions of paddy soils may exert different bioavailability when exposure to soil fauna. Thus, content and chemical extractability may not be appropriate index for assessing the potential environmental risk and bioavailability to soil fauna for persistent organic pollutants in paddy soils.

Animals↗