Residues effects of isoproturon in mature earthworm (Aporrectodea caliginosa) under laboratory conditions

This study was conducted to investigate the residues of isoproturon and its metabolites, 1-(4-isopropylphenyl)-3-methylurea, 1-(4-isopropylphenyl) urea, and 4-isopropylanilin in soil and mature earthworms under laboratory conditions. Mature earthworms (Aporrectodea caliginosa) were exposed for various durations (7, 15, 30, and 60 days) to soils contaminated with isoproturon concentrations (2, 4, 6, 8, and 10 mg.kg(-1) soil). The decrease in isoproturon concentration in the soil depended on initial concentration it was slower at higher concentrations. The isoproturon and its metabolites accumulated in earthworms it increased during the first 15 days and decreased thereafter. Acute toxicity of isoproturon was determined together with total soluble protein content and glycogen of worms. These parameters were related to isoproturon concentration in soil and earthworms. No lethal effect of isoproturon was observed even at the concentration 1000 mg.kg(-1) soil after 60 days of exposure. A reduction of total soluble protein was observed in all treated worms (maximum 59.54%). This study is suggesting the use of the total soluble protein content and glycogen of earthworms as biomarker of exposure to isoproturon.     

Earthworms as paratenic hosts of toxoplasmosis in eastern barred bandicoots in Tasmania

An experimental feeding study was designed to assess the role of earthworms in the transmission of Toxoplasma gondii infection to eastern barred bandicoots (Perameles gunnii). Six animals with no agglutinating antibodies to T. gondii were fed artificially cultured earthworms that had been maintained in autoclaved nutrient-enriched soil. Two animals were given earthworms that had been maintained in soil contaminated with T. gondii oocysts (P89/VEG strain); two animals were fed on earthworms, which initially had been exposed to soil containing T. gondii oocysts then transferred through three changes of sterile soil; two control bandicoots were fed earthworms maintained in sterile soil. Both bandicoots fed earthworms maintained in T. gondii contaminated soil died 11 and 14 days after feeding. The necropsy findings were consistent with acute toxoplasmosis. Bandicoots fed earthworms exposed to oocysts but then transferred through changes of sterilized soil remained healthy as did control animals. All surviving animals remained seronegative over the 6 wk observation period after feeding. These findings confirm that earthworms, a major component of the natural diet of P. gunnii, can transmit T. gondii infection. It appears that oocysts present in the alimentary tracts of the worms, rather than infective stages of T. gondii in worm somatic tissues, are responsible for these infections.     

Earthworm survival in oil contaminated soil

Earthworms are an important component of the soil biota and their response to oil pollution needs to be better understood. Laboratory investigations were undertaken to determine the concentrations of crude oil in soil that leads to death of Lumbricus terrestris and Eisenia fetida and to determine the propensity of L. terrestris to move away from contaminated soil. Clemville sandy clay loam was amended to contain maximum oil contents of 1.5 – 2.5% depending on the particular experiment. Additionally, the ability of L. terrestristo survive in bioremediated oil-contaminated soil was evaluated. An oil content of 0.5% was not harmful to survival of earthworms for 7 d but an oil concentration of 1.5% reduced survival to less than 40%. Bioremediated soil containing 1.2% oil did not reduce survival of L. terrestrisduring 10 d. Survival of L. terrestrisin unweathered oil was improved when free movement between contaminated and uncontaminated soil was possible. Casts of earthworms exposed to oil-containing soil contained 0.2% total petroleum hydrocarbons. An allowable regulatory level of 1% oil contamination in soil may not allow for survival of earthworms.     

Evaluation for Hsp70 as a biomarker of effect of pollutants on the earthworm Lumbricus terrestris

Induction of heat shock proteins (Hsps) is often associated with a cellular response to a harmful stress or to adverse life conditions. The main aims of the present study were (1) to assess if stress-induced Hsp70 could be used to monitor exposure of the earthworm species Lumbricus terrestris to various soil pollutants, (2) to assess the specificity of pollutants in their tissue targeting and in Hsp70 induction, and (3) to evaluate if dose-response relationships could be established and if the stress-response observed was specific. The midgut/intestinal tissues of L. terrestris are shown to express an inducible member of the Hsp70 family after heat shock treatment in vitro and exposures to different soil toxicants in vivo (re: artificial soil). Short-term (24-72 hours) and long-term (14-16 days) exposures to the chemical standards chloroacetamide and pentachlorophenol and to heavy metals (Pb++, Cd++, Cu++, and Hg++) also affected the earthworms, and Hsp70 was induced in their midgut/intestinal tissues. After a 3-day exposure to heavy metals, the level of Hsp70 induction in the midgut/intestinal tissues appears to correlate well with the reported in vivo and in vitro toxicity data. Comparatively, in proximal and midbody wall muscle tissues of animals exposed to the heavy metals, a decrease in expression of Hsp70 was sometimes detected. Thus Hsp analysis by Western blot in L. terrestris tissues and particularly in the midgut/intestine proved to be a suitable and sensitive assay for adverse effects in earthworms and showed a good level of reproducibility despite some individual variations. The use of pristine/nonexposed animals transposed into contaminated environments as in the present study should therefore be of high ecological relevance. Induction of Hsp70 in earthworms should represent not only a good wide-spectrum biomarker of exposure but also a biomarker of effect since known toxicants altered gene expression in tissues of these animals, as contrasted with a simple accumulation of Hsp. Hence, the detection of Hsp70 in earthworms can constitute an early-warning marker for the presence of potentially deleterious agents in soils, with L. terrestris in particular and earthworms in general acting as potential sentinel animal species.     

Glycogen-lead relationship in the earthworm Dendrobaena rubida from a heavy metal site

Control individuals contained no lead in the chloragocytes but high alpha-glycogen rosette reserves. Starvation of contaminated earthworms for 4d caused a lead loss and the chlorgocytes possessed fewer debris vesicles than those of unstarved worms, suggesting that the debris vesicles may be the route for at least some of the lead loss. No glycogen deposits were observed in the chloragocytes of starved or unstarved earthworms from contaminated soil. Maintenance of contaminated earthworms in potting compost caused lead losses similar to those sustained by starvation, but the chloragocyte cytoplasm possessed beta-glycogen reserves. Specimens maintained in lead-spiked potting compost showed lead levels similar to those of earthworms taken directly from contaminated soil. No beta-glycogen accumulations were observed under this enriched regime. Although the possible interference of lead in carbohydrate metabolism is discussed, the results do not wholly support metabolic inhibition by lead. It is hypothesised that lead sequestration is energy-demanding and that in the absence of an energy-rich diet glycogen stores fail to accumulate. In the presence of an organic-rich medium, elevated lead levels preclude glycogen formation, because of the high sequestration-demand, but at lower lead levels beta-glycogen deposits occur if a high organic diet is available.     

Genotoxicity assessment in Eisenia andrei coelomocytes: a study of the induction of DNA damage and micronuclei in earthworms exposed to B[a]P- and TCDD-spiked soils

Earthworms are useful indicators of soil quality and are widely used as model organisms in terrestrial ecotoxicology. The assessment of genotoxic effects caused by environmental pollutants is of great concern because of their relevance in carcinogenesis. In this work, the earthworm Eisenia andrei was exposed for 10 and 28 days to artificial standard soil contaminated with environmentally relevant concentrations of benzo[a]pyrene (B[a]P) (0.1, 10, 50ppm) and 2,3,7,8-tetrachloro-dibenzo-para-dioxin (TCDD) (1×10(-5), 1×10(-4), 2×10(-3)ppm). Micronucleus (MNi) induction was evaluated in earthworm coelomocytes after DNA staining with the fluorescent dye DAPI. In the same cells, the DNA damage was assessed by means of the alkaline comet assay. Induction of MNi in coelomocytes, identified according to standard criteria, was demonstrated. B[a]P exposure for 10 and 28 days induced a significant increase in MNi frequency. In TCDD-treated earthworms, a significant effect on chromosomal damage was observed at all the concentrations used; surprisingly, greater effects were induced in animals exposed to the lowest concentration (1×10(-5)ppm). The data of the comet assay revealed a significant increase in the level of DNA damage in coelomocytes of earthworms exposed for 10 and 28 days to the different concentrations of B[a]P and TCDD. The results show that the comet and MN assays were able to reveal genotoxic effects in earthworms exposed even to the lowest concentrations of both chemicals tested here. The combined application in E. andrei of the comet assay and the micronucleus test, which reflect different biological mechanisms, may be suggested to identify genotoxic effects induced in these invertebrates by environmental contaminants in terrestrial ecosystems.     

蚯蚓-秸秆及其交互作用对黑麦草修复Cu污染土壤的影响

以高沙土为供试土壤,加入Cu^2＋以模拟成：0,100,200,400mg／kgCu^2＋的Cu污染土壤,设置接种蚯蚓（E）、表施秸秆（M）,同时加入蚯蚓和秸秆（ME）及不加蚯蚓和秸秆的对照（CK）4个处理,并种植黑麦草。研究蚯蚓、秸秆相互作用对黑麦草吸收、富集铜的影响。结果表明：加入秸秆显著提高了蚯蚓的生物量,一定程度上缓解了重金属对蚯蚓的毒害,同时蚯蚓显著提高了秸秆的分解率,较无蚯蚓对照提高了58．11％～77．32％。接种蚯蚓（E,ME）还提高了土壤有效态重金属（DTPA-Cu）含量,秸秆处理（M）则降低了土壤有效态重金属含量。研究还发现,E处理促进了黑麦草地上部生长,而M和ME处理均显著提高了黑麦草地下部的生物量。E和ME处理同时提高了植物地上部和地下部的Cu浓度及Cu吸收量,M处理则只对植物的地下部Cu浓度和Cu吸收量有显著促进作用。总体来看,E处理、M处理及ME处理分别使黑麦草地上部Cu富集系数提高了31．22％～121．07％．2．12％～61．28％和25．56％～132．64％。     

蚯蚓作用下污泥重金属形态变化及其与化学生物学性质变化的关系

城市污泥处理是一项世界性难题,污泥农业利用是其最简单有效的资源化利用方式之一,但污泥中较高的重金属含量限制了其实际推广应用,利用蚯蚓-超富集植物联合修复污泥重金属的方法已引起国内外研究者的关注。以新鲜城市脱水污泥为研究对象,接种赤子爱胜蚓(Eisenia fetida)进行室内培养试验,系统研究蚯蚓作用下污泥重金属形态的变化,及其与污泥氧化还原条件、化学和微生物性质变化的关系,以期为蚯蚓-超富集植物联合修复技术在污泥重金属处理中的应用提供理论依据。结果表明,试验前期蚯蚓在污泥中能正常生长和存活,前20 d总生物量增加了52%。蚯蚓可以显著促进污泥中的Cu、Zn、Cd、Ni等重金属从残渣态和铁锰态等稳定形态向交换态和水溶态等有效形态转化。还可以显著降低污泥中还原性物质的含量,减缓p H值下降速度,降低总有机碳含量,促进铵态氮向硝态氮转化,减少污泥微生物的数量并增加其种群活性。蚯蚓作用下,污泥中重金属的活化程度与还原性物质的含量呈显著负相关,而与微生物种群的活性呈显著正相关(P0.05)。综上所述,蚯蚓可以促进污泥重金属的活化,并改善污泥的肥力条件,为修复植物在污泥中的正常生长和对重金属离子的快速吸收提供有利条件。     

The role of proteolytic enzymes in protein degradation during senescence of rice leaves

The role of proteolytic enzymes in protein degradation of detached and intact leaves of rice seedling ( Oryza sativa L. cv. Taiching Native 1) during senescence and of mature leaves during reproductive development was investigated. The amount of soluble protein decreased by about 50% in 2, 4, and 15 days for detached, intact and mature leaves, respectively. Three proteolytic enzyme activities were monitored with pH optima of 4.5 for hemoglobin-digesting proteinase, 5.5 for carboxypeptidase and 8.0 for aminopeptidase. No azocoll-digesting proteinase activity could be detected in rice leaves. Dialysis did not alter the activities of any of the three proteolytic enzymes. Acid proteinase activity and aminopeptidase activity were highly unstable during storage of the enzyme extracts at 4°C. Proteolysis was stimulated by inclusion of meroaptoethanal either in the extraction medium or the assay medium.
Acid proteinase, carboxypeptidase and aminopeptidase were all present in detached, intact and mature leaves throughout senescence. There seems to be a direct correlation between protein degradation and increases of acid proteinase and carboxypeptidase activity in seedling leaves (detached and intact) during senescence. In senescing (detached and intact) leaves of seedlings the acid proteinase activity developed first, while that of carboxypeptidase developed later. Acid proteinase and carboxypeptidase may play major roles in protein degradation of leaves from seedlings during senscence. During reproductive development, protein degradation was associated with decreases in the activities of acid proteinase, carboxypeptidase and aminopeptidase in mature leaves suggesting that the enzymes were less important for protein degradation in this system. Hence, the role of protelytic enzymes in protein degradation during senescence of rice leaves appears to depend largely on the leaf system used.     

Competition alters plant species response to nickel and zinc

Phytoextraction can be a cost-efficient method for the remediation of contaminated soils. Using species mixtures instead of monocultures might improve this procedure. In a species mixture, an effect of heavy metals on the species' performance can be modified by the presence of a co-occuring species. We hypothesised that (a) a co-occuring species can change the effect of heavy metals on a target species, and (b) heavy metal application may modifiy the competitive behaviour between the plants. We investigated these mechanisms in a greenhouse experiment using three species to serve as a model system (Carex flava, Centaurea angustifolia and Salix caprea). The species were established in pots of monocultures and mixtures, which were exposed to increasing concentrations of Ni and Zn, ranging from 0 to 2,500 mg/kg. Increased heavy metal application reduced the species' relative growth rate (RGR); the RGR reduction being generally correlated with Ni and Zn concentrations in plant tissue. S. caprea was an exception in that it showed considerable Zn uptake but only moderate growth reduction. In two out of six cases, competitors significantly modified the influence of heavy metals on a target species. The interaction can be explained by an increased uptake of Zn by one species (in this case S. caprea) that reduced the negative heavy metal effect on a target species (C. flava). In two further cases, increasing heavy metal application also altered competitive effects between the species. The mechanisms demonstrated in this experiment could be of relevance for the phytoextraction of heavy metals. The total uptake of metals might be maximised in specific mixtures, making phytoextraction more efficient.     

Role of earthworm-straw interactions on phytoremediation of Cu contaminated soil by ryegrass

It is well known that the earthworm's activities can increase the availability of soil nutrients, improve soil structure, and enhance the biomass of plants in uncontaminated soil. Recently, many researchers found that some metal-tolerant earthworms can survive and even change the fractional distribution of heavy metals in contaminated soil. Furthermore, it has been revealed that earthworms are able to increase metal availability, and therefore, accumulate more metals in plants through their burrowing and casting activity. It is clear that the influence of soil animals is an important factor for phyto-remedation that must be taken into account. ~In this article, the authors studied some effects of addition of earthworms (Metaphire guillelmi), corn straw, and in combinations of earthworms and corn straw on the growth and Cu uptake by ryegrass in Cu contaminated pot soils. The experiment consisted of four levels of Cu addition (0, 100, 200, 400 mg·kg?1) and four treatments. The treatments were 1. control (CK); 2.straw mulching only (M); 3. earthworm additions to soil only (E); and 4.straw mulching plus earthworm additions (ME). Each treatment had three replicates. 10 seeds of ryegrass (Lolium multiflorum) were sowed in each pot and harvested after 30 days. After 30 days of incubation, all earthworms were found to be alive and the pot soils were burrowed through by earthworms. Results showed that the biomass of earthworm declined with the increase of the dosage of Cu additions. The biomass of earthworm increased significantly in treatment 4 (ME) as compared with treatment 3 (E). Not only the earthworms could get more food from the straw, but also could counteract some negative effects of Cu on the earthworms. The rates of straw decomposition in ME treatment increased by about 58.11% ?77.32%. The earthworm activities increased root biomass of ryegrass significantly, but did not show the effect on plant root growth. On the contrary, straw enhanced roots biomass significantly instead of shoots biomass. It was also found that the concentration of Cu in the plant shoot and the plant root, as well as plant Cu uptake were enhanced by earthworm's activities and straw mulching. The increased amount by straw mulching was lower than that of earthworms (E). The treatment of the earthworm–straw mulching combinations enhanced plant Cu concentration, and the amount increased by it was lower than that of the earthworm treatment (E) but higher than that of straw mulching treatment (M). The accumulation factors of copper in the shoots of ryegrass were increased by 31.22% ?121.07%, 2.12% ?61.28% and 25.56% ?132.64%, respectively, in treatment 3(E), 2(M), and 4(ME), respectively. In conclusion, the earthworm activities, straw-mulching and their interactions may have potential roles in elevating phyto-extraction efficiency in low to medium level Cu contaminated soil.     

The collembola Lobella sokamensis juvenile as a new soil quality indicator of heavy metal pollution

We propose Lobella sokamensis Deharveng and Weiner, 1984 juvenile as a new soil quality indicator to assess heavy metal polluted soils. L. sokamensis is a collembola commonly found with earthworms, and it plays a key role in decomposing dead earthworms. The soil quality assessment of cadmium (Cd), copper (Cu), antimony (Sb), and zinc (Zn) on the survival of L. sokamensis adults were performed in artificial soil. The LC50 values for L. sokamensis adults exposed to Cd, Cu, Sb and Zn for 5 days were calculated to be 4729, 4472, 4702, and 2521 mg kg⁻¹, respectively. L. sokamensis juveniles (10–12 days) were also exposed to Cd, Cu, Sb, and Zn, and 5 days-LC50 values for Cd, Cu, Sb, and Zn were 282, 229, 447, and 163 mg kg⁻¹, respectively. Both adult and juvenile survivals of L. sokamensis in metal spiked soil were inhibited. We found that the soil quality assessment using L. sokamensis juveniles were sensitive enough to evaluate soil pollution, compared to conventional Folsomia candida assay which is widely used. In addition, L. sokamensis has advantage for easy detection in soil due to the big and orange-colored body. The 5 days-soil quality assessment using L. sokamensis juveniles appears to be rapid protocols for the collembola assay. This is the first report on the collembola assay of heavy metal contaminated soils using the L. sokamensis as a soil indicator.     

From industrial sites to environmental applications with <Emphasis Type="Italic">Cupriavidus metallidurans</Emphasis>

Cupriavidus metallidurans CH34 and related strains are adapted to metal contaminated environments. A strong resistance to environmental stressors and adaptation make it ideal strains for survival in decreasing biodiversity conditions and for bioaugmentation purposes in environmental applications. The soil bacterium C. metallidurans is able to grow chemolithoautotrophically on hydrogen and carbon dioxide allowing a strong resilience under conditions lacking organic matter. The biofilm growth on soil particles allows coping with starvation or bad conditions of pH, temperature and pollutants. Its genomic capacity of two megaplasmids encoding several heavy metal resistance operons allowed growth in heavy metal contaminated habitats. In addition its specific siderophores seem to play a role in heavy metal sequestration besides their role in the management of bioavailable iron. Efflux ATPases and RND systems pump the metal cations to the membrane surface where polysaccharides serve as heavy metal binding and nucleation sites for crystallisation of metal carbonates. These polysaccharides contribute also to flotation under specific conditions in a soil-heavy metals–bacteria suspension mixture. An inoculated moving bed sand filter was constructed to treat heavy metal contaminated water and to remove the metals in the form of biomass mixed with metal carbonates. A membrane based contactor allowed to use the bacteria as well in a versatile wastewater treatment system and to grow homogeneously formed heavy metal carbonates. Its behaviour toward heavy metal binding and flotation was combined in a biometal sludge reactor to extract and separate heavy metals from metal contaminated soils. Finally its metal-induced heavy metal resistance allowed constructing whole cell heavy metal biosensors which, after contact with contaminated soil, waste, solids, minerals and ashes, were induced in function of the bioavailable concentration (Cd, Zn, Cu, Cr, Co, Ni, Tl, Pb and Hg) in the solids and allowed to investigate the speciation of immobilization of those metals. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Characterization and cDNA cloning of midgut carboxypeptidases from Trichoplusia ni

Carboxypeptidase A and carboxypeptidase B activities from the midgut of Trichoplusia ni larvae were characterized. In the T. ni larval midgut, the primary digestive carboxypeptidase activity was attributed to carboxypeptidase A, which was eight times more active than carboxypeptidase B. Both the midgut carboxypeptidase A and carboxypeptidase B exhibited maximal activities at pH 8.0-8.5 and were similarly susceptible to inhibition by potato carboxypeptidase inhibitor and phenanthroline. The midgut carboxypeptidase activities were analyzed in T. ni larvae fed on various diet sources and the results indicated that midgut carboxypeptidase activities per milligram of gut were similar regardless of the amount of dietary proteins or amino acids. However, midgut carboxypeptidase A activity was significantly higher in larvae exposed to soybean trypsin inhibitor and was significantly lower in larvae fed on broccoli foliage. From the T. ni larval midgut, five putative carboxypeptidase cDNAs were cloned, demonstrating that midgut carboxypeptidase activities are composed of multiple carboxypeptidase types. Sequence analysis indicated that the midgut carboxypeptidases were produced as secreted proenzymes which could be activated after removal of an N-terminal activation fragment by a trypsin. Two cloned cDNAs are predicted to code for carboxypeptidase A and one cDNA is predicted to code for a putative carboxypeptidase B. The other two cDNAs are highly similar to carboxypeptidase A and carboxypeptidase B in sequences, but their activity was not predictable.     

蚯蚓对土壤-植物系统生态修复作用研究进展

土壤-植物系统是生物圈的基本结构单元,土壤与植物之间存在密切的相互反馈.土壤退化导致植物面临各种非生物胁迫,植物的生理代谢遭到干扰,养分获取受到抑制.蚯蚓被称为"生态系统的工程师".蚯蚓能够通过调控土壤物理-化学-生物学特性,改良退化土壤(盐碱土、重金属和有机污染物污染土壤),缓解植物所受胁迫,增加土壤养分有效性,促进...     

Purification, cDNA cloning, and expression of a new human blood plasma glutamate carboxypeptidase homologous to N-acetyl-aspartyl-alpha-glutamate carboxypeptidase/prostate-specific membrane antigen

We describe the identification, cDNA cloning, and biochemical characterization of a new human blood plasma glutamate carboxypeptidase (PGCP). PGCP was co-purified from human placenta with lysosomal carboxypeptidase, cathepsin A, lysosomal endopeptidase, cathepsin D, and a gamma-interferon-inducible protein, IP-30, using an affinity chromatography on a Phe-Leu-agarose column. A PGCP cDNA was obtained as an expressed sequence tag clone and completed at 5'-end by rapid amplification of cDNA ends polymerase chain reaction. The cDNA contained a 1623-base pair open reading frame predicting a 541-amino acid protein, with five putative Asn glycosylation sites and a 21-residue signal peptide. PGCP showed significant amino acid sequence homology to several cocatalytic metallopeptidases including a glutamate carboxypeptidase II also known as N-acetyl-aspartyl-alpha-glutamate carboxypeptidase or as prostate-specific membrane antigen and expressed glutamate carboxypeptidase activity. Expression of the PGCP cDNA in COS-1 cells, followed by Western blotting and metabolic labeling showed that PGCP is synthesized as a 62-kDa precursor, which is processed to a 56-kDa mature form containing two Asn-linked oligosaccharide chains. The mature form of PGCP was secreted into the culture medium, which is consistent with its intracellular localization in secretion granules. In humans, PGCP is found principally in blood plasma, suggesting a potential role in the metabolism of secreted peptides.     

Glycogen-lead relationship in the earthworm Dendrobaena rubida from a heavy metal site

Summary Control individuals contained no lead in the chloragocytes but high -glycogen rosette reserves. Starvation of contaminated earthworms for 4d caused a lead loss and the chloragocytes possessed fewer debris vesicles than those of unstarved worms, suggesting that the debris vesicles may be the route for at least some of the lead loss. No glycogen deposits were observed in the chloragocytes of starved or unstarved earthworms from contaminated soil. Maintenance of contaminated earthworms in potting compost caused lead losses similar to those sustained by starvation, but the chloragocyte cytoplasm possessed -glycogen reserves. Specimens maintained in lead-spiked potting compost showed lead levels similar to those of earthworms taken directly from contaminated soil. No -glycogen accumulations were observed under this enriched regime.Although the possible interference of lead in carbohydrate metabolism is discussed, the results do not wholly support metabolic inhibition by lead. It is hypothesised that lead sequestration is energy-demanding and that in the absence of an energy-rich diet glycogen stores fail to accumulate. In the presence of an organic-rich medium, elevated lead levels preclude glycogen formation, because of the high sequestration-demand, but at lower lead levels -glycogen deposits occur if a high organic diet is available.     

The occurrence and localisation of heavy metals and glycogen in the earthwormsLumbricus rubellus andDendrobaena rubida from a heavy metal site

Summary The lead content of whole earthworms, highest in contaminated site specimens, was significantly higher inDendrobaena rubida thanLubricus rubellus and a species difference in zinc was also recorded. Selective feeding and differential absorption are discounted and a species difference in maximum tolerance to body lead is suggested. Copper was low in all specimens. Chloragocytes and intestinal tissue showed significantly higher lead levels in contaminated earthworms than in control material. Ultrastructurally, chlorgocytes from contaminated earthworms showed electron dense flecks associated with the chroragosome peripheries and within the debis vesicles. Very fine flecks occurred in the nuclei, but mitochondria and Golgi were indistinguishable from control material. Preliminary X-ray microanalysis of contaminated chloragocytes revealed lead and phosphorus. The deposits within the chloragocytes might represent unbound lead precipitated by phosphate buffer; flecks being absent from contaminated, citrate buffered material and from control material. The chloragosomes are proposed as possible sites for sequestered lead.Chloragocyte and intestinal glycogen levels were significantly higher in control material where the chloragocyte cytoplasm was rich in -glycogen rosettes, these being absent from lead contaminated cells. The glycogen-lead correlation suggests that the metabolism of contaminated chloragocytes is directed towards lead sequestration, though differing nutritional states cannot be ignored.The assistance of Mr. P.M. Webster, University of Keele, is gratefully acknowledged.