Heavy metals in the terrestrial isopod Porcellio scaber Latreille. II. Subcellular fractionation of metal-accumulating lysosomes from hepatopancreas

Two populations of metal contaminated Porcellio scaber Latreille were studied: one consisting of animals that had been fed heavy metals in the laboratory for several months, and one from a metal-polluted site in the field (Braubach, FRG). Density gradient centrifugation was performed on hepatopancreas homogenates in order to identify cellular fractions and their association with lead, copper and cadmium. Marker enzymes were used for localisation of cellular fractions in the density gradient. Two lysosomal fractions, called the light and heavy fraction, were separated. They contained mainly lead, but also copper and some cadmium.Abbreviations AP acid phosphatase - ER endoplasmic reticulum - LDH lactic dehydrogenase - MDH malic dehydrogenase - NADH nicotinamide adenine dinucleotide - P. scaber Porcellio scaber     

Early reproduction and increased reproductive allocation in metal-adapted populations of the terrestrial isopod Porcellio scaber

Organisms inhabiting metal-contaminated areas can be stressed by metal exposure and are possibly subject to selection, resulting in increased metal tolerance and changes in growth and/or reproduction characteristics. In a previous study it was found that in the terrestrial isopod Porcellio scaber, sampled from the vicinity of a zine smelter, the body size was small and the brood size was large compared to isopods from a reference area. To assess whether these differences were due to genetic differentiation between strains, isopods collected from a reference wood, a zinc smelter area and a lead mine were cultured on non-polluted food, while growth, reproduction and metal concentrations were studied in first and second laboratory generations. The isopods from the three populations differed in age and weight at first reproduction, although there were hardly any differences in growth. The females of the mine and the smelter population started to reproduce earlier, at a lower weight, which resulted in fewer young per female. However, reproductive allocation (=wight of young relative to the weight of the mother) was higher in mine and smelter isopods. We conclude that the isopods at the metal-contaminated sites have been selected for early reproduction and increased reproductive allocation. The results indicate that populations inhabiting metal-polluted sites have probably undergone evolutionary changes. This study showed that growth and reproduction characteristics of different populations under laboratory conditions may provide information on selection processes in the field.     

Uropod and lateral plate glands of the terrestrial isopod Porcellio scaber Latr. (Oniscidae,Crustacea): An ultrastructural study

Lateral plate and uropod glands are composed of a binucleated gland cell, a ramified intermediate cell, and an elongated duct cell. The gland cell is divided into several lobes and forms numerous short processes in its periphery. The cytoplasm contains many secretory granules. The granules release their content into the intercellular collecting ducts between the gland cell and the branched extensions of the intermediate cell. The collecting ducts merge into a funnel-shaped space surrounded by the intermediate cell. The duct cell is lined by a cuticular intima and contains long striated bundles of fibrils. The duct cell consists of two different regions. The proximal region is characterized by microvilli on the luminal side and contains many organelles. In the distal region microvilli are lacking and organelles are scarce. Structurally, the uropod and lateral plate glands differ in the number of components within the granules. This is in accordance with the differences in the characteristics of the secretory products of the two gland types. The morphology of the glands, particularly the peripheral position of the collecting system, is unique among exocrine glands of arthropods. J. Morphol. 233:183–193, 1997 © 1997 Wiley-Liss, Inc.     

An intra-species demonstration of the independence of distance and time in turn alternation of the terrestrial isopod, Porcellio scaber

In a study of turn alternation in the terrestrial isopod Porcellio scaber (woodlouse), smaller individuals (<11 mm long) took significantly longer to traverse a 60-mm open-ended post-forced-turn runway following a 90 degrees forced turn, than larger subjects (>11 mm long). However, there were no significant differences between the two groups in their probability of alternating, or in the magnitude of alternating turns as reflected in the size of the free-turn angles they turned on emerging from the runway. Nor was there a significant relationship for either group between the time taken to traverse the runway and either the probability of alternating or the size of a free-turn angle. In a second experiment, significant alternation occurred after woodlice emerged from runways that were 30 and 60 mm long, but not when they were 100 or 145 mm long. Their free-turn angles also became smaller as the runway length increased. Overall, the results of the three experiments supported the long-held view that, for reasons not yet understood, distance and time cannot necessarily be equated in the determination of invertebrate turn alternation.     

Spatial distribution of calcite and amorphous calcium carbonate in the cuticle of the terrestrial crustaceans Porcellio scaber and Armadillidium vulgare

The crustacean cuticle is an interesting model to study the properties of mineralized bio-composites. The cuticle consists of an organic matrix composed of chitin–protein fibres associated with various amounts of crystalline and amorphous calcium carbonate. It is thought that in isopods the relative amounts of these mineral polymorphs depend on its function and the habitat of the animal. In addition to the composition, the distribution of the various components should affect the properties of the cuticle. However, the spatial distribution of calcium carbonate polymorphs within the crustacean cuticle is unknown. Therefore, we analyzed the mineralized cuticles of the terrestrial isopods Armadillidium vulgare and Porcellio scaber using scanning electron-microscopy, electron probe microanalysis and confocal μ-Raman spectroscopic imaging. We show for the first time that the mineral phases are arranged in distinct layers. Calcite is restricted to the outer layer of the cuticle that corresponds to the exocuticle. Amorphous calcium carbonate is located within the endocuticle that lies below the exocuticle. Within both layers mineral is arranged in rows of granules with diameters of about 20 nm. The results suggest functional implications of mineral distribution that accord to the moulting and escape behaviour of the animals.     

Digestion of prey in foraminifera is not anomalous: A correlation of light microscopic, cytochemical, and hvem technics to study phagotrophy in two allogromiids

Correlative light, high-voltage electron and conventional electron microscopic methods were used to investigate digestion in two allogromiid foraminiferans, Allogromia sp., strain NF, and A. laticollaris Arnold. Microscopic observations showed that bacterial prey are phagocytosed by reticulopodia and are transported to the allogromiid cell body within blister-like phagosomes. Larger prey (algae, diatoms) are transported along the reticulopodial surface and are either stored extrathalamously or phagocytosed at the oral opening (peduncle). Studies of allogromiids optimally fixed and labeled with an extracellular-space label (colloidal thorium) showed that phagocytosed prey are completely enclosed by a plasma membrane envelope; this finding was corroborated by a serial-section three-dimensional reconstruction of the oral zone of one allogromiid. Cytochemical staining for acid phosphatase showed that lysosomes are absent from reticulopods but abundant in the cell body, particularly in the oral zone cytoplasm. We conclude that digestion in allogromiid foraminiferans is accomplished by a vacuole-based digestive apparatus and not by extracellular digestion within a lacunary system, as has been suggested in earlier studies.     

Membrane particle distribution in the sternal epithelia of the terrestrial isopod Porcellio scaber latr. (Crustacea, oniscidea) during CaCO(3) deposit formation and resorption, a freeze-etch analysis.

The anterior sternal epithelium of terrestrial isopods transports cuticular Ca(2+) to and from large sternal CaCO(3) deposits. We analyzed the anterior and posterior sternal epithelium by the means of the freeze-etch technique and measured the size distribution and density of intramembrane particles (IMPs) during three different molting stages. At least three IMP size classes around 4.5, 7.7, and 9.4 nm can be distinguished on the P-face of the apical and basolateral plasma membrane. An additional size class of around 12.8 nm is restricted to the apical compartment. In the anterior sternal epithelium, the density of these large particles changes by a factor of 1.9 during the molt cycle, suggesting a role in CaCO(3) formation and/or resorption. The density of the smaller IMPs rises transiently by a factor of 1.3 in the posterior sternal epithelium only. The IMP density of the basolateral plasma membrane increases significantly by a factor of 1.4 and 1.3 in the anterior and posterior sternal epithelia, respectively. The results indicate that increases in the IMP density contribute to the differentiation to an increased transport activity during the cyclic enlargements of the plasma membrane surface area in the anterior sternal epithelium.     

Light and Electron Microscope Study of Dunaliella primolecta Butcher (Volvocida)*

SYNOPSIS. Light and electron microscope observations on Dunaliella primolecta Butcher from logarithmic and stationary phases of batch cultures are correlated. Except for the lack of a cell wall the fine structure has typical volvocid features. The transition from logarithmic to stationary phase is marked by changes in content and size of cytoplasmic vacuoles, accumulation of cytoplasmic lipid, accumulation of starch in the plastid matrix, and by the formation of autophagosome-like bodies. The organelles in stationary-phase flagellates are closely packed together because of the cytoplasmic lipid and starch-distended chloroplast. Organisms from logarithmic phase have an abundant ribosome-packed groundplasm supporting the organelles. In the cytochemical demonstration of acid phosphatase activity, Golgi cisternae and smooth and coated Golgi vesicles contain Gomori reaction product. The possible roles of the Golgi apparatus in this flagellate are discussed.     

Mutations Affecting the Trichocysts in Paramecium aurelia. I. Morphology and Description of the Mutants*

Six types of genic mutants have been isolated. Their phenotypes range from animals with no trichocysts (trichless), to animals with morphologically abnormal trichocysts (football, stubby, pointless, screwy-cigar), to animals which are incapable of extruding otherwise normal looking trichocysts (nondischarge). The football mutant possesses football-shaped trichocysts, which, unlike wild-type trichocysts, do not attach at the cortex. The stubby mutant possesses shorter trichocysts which have a very highly variable morphology. The screwy-cigar animals have thinner and usually longer trichocysts than those found in wild-type cells. The trichocysts of the pointless mutant have all the components of the wild-type organelles but not in their proper relationship. Electron microscopic studies of the mutants have demonstrated that although the morphology of the various mutant trichocysts may differ, their ultrastructure and early developmental stages are comparable to those of trichocysts found in wild type. The mutations are usually pleiotropic, affecting other systems besides trichocysts. The existence of these mutants, particularly trichless, poses some interesting questions regarding the function of trichocysts, and also gives insight into the development of trichocysts.     

Comparison of phosphatase localization in the intraradical hyphae of arbuscular mycorrhizal fungi,Glomus spp. and Gigaspora spp.

The localization of acid and alkaline phosphatases in the intraradical hyphae of the arbuscular mycorrhizal fungi, Glomus mosseae (Nicol. and Gerd.) Gerd. and Trappe (Gm), Gl. etunicatum Becker and Gerd. (Ge) and Gigaspora rosea Nicol. and Schenck (Gir) were compared. Marigold (Tagetes patula L.) and leek (Allium porrum L.) were inoculated with each of the three fungi. The mycorrhizal roots were harvested at 3, 4, 5 and 6 weeks after sowing (WAS), treated with a digestion solution containing cellulase and pectinase, and then stained for phosphatase activities at pH 5.0 and pH 8.5. The development of fungal structures in the host root was also examined. Gm formed fine-branched (mature) arbuscules only at the early phase of infection (3 to 4 WAS). Mature arbuscules of Ge and Gir were observed from the early phase (4 WAS) up to the end of experiment. At pH 5.0, the localization of the phosphatase activities of the three fungi were similar irrespective to host plant species. The activity appeared in mature arbuscules and intercellular hyphae, whereas the collapsed arbuscules were inactive. Ten millimolar NaF, an acid phosphatase inhibitor, inhibited the phosphatase activities of Gm and Ge but did not affect that of Gir. At pH 8.5, a difference among the fungal species was found in the localization of phosphatase activity while that between host species was not. The mature arbuscules were also the active sites in all three species. Only Gir showed the activity in the intercellular hyphae while the two Glomus spp. did not. Five millimolar EDTA inhibited the activity of Gir at pH 8.5 while the activities of Ge and Gm were not affected by either 5 mM EDTA or 10 mM KCN (both are alkaline phosphatase inhibitors).     

上海市崇明岛城镇河流沉积物重金属累积与环境风险

小城镇河流沉积物重金属累积状况间接指示了河流水环境质量现状,研究河流沉积物重金属累积水平与环境风险具有重要的现实意义.对上海市崇明岛崇南分区3个典型城镇(城桥镇、堡镇、陈家镇)河流沉积物重金属累积特征进行了研究.结果表明,3个城镇河流沉积物中Zn、Cr、Cu、Pb和Cd平均含量分别为：城桥镇103.1、79.2、39.0、29.7和0.300 mg·kg^-1;堡镇99.0、104.0、50.5、32.7和0.308 mg·kg^-1;陈家镇86.7、79.5、32.2、30.5和0.274 mg·kg^-1;崇南分区94.72、81.35、38.52、30.78和0.297 mg·kg^-1.用地累积指数法(I_geo)对沉积物重金属进行的环境风险评估结果为：崇明3个城镇河流沉积物重金属均为低-中等环境风险,其中3个城镇达到中等环境风险的概率为城桥镇>堡镇>陈家镇.     

Purple acid phosphatase-like sequences in prokaryotic genomes and the characterization of an atypical purple alkaline phosphatase from Burkholderia cenocepacia J2315

Purple acid phosphatases (PAP) are a group of dimetallic phosphohydrolase first identified in eukaryotes. Bioinformatics analysis revealed 57 prokaryotic PAP-like sequences in the genomes of 43 bacteria and 4 cyanobacteria species. A putative PAP gene (BcPAP) from the bacteria Burkholderia cenocepacia J2315 was chosen for further studies. Synteny analysis showed that this gene is present as an independent gene in most of the members of the genus Burkholderia. The predicted 561 a.a. polypeptide of BcPAP was found to harbour all the conserved motifs of the eukaryotic PAPs and an N-terminal twin-arginine translocation signal. Expression and biochemical characterization of BcPAP in Escherichia coli revealed that this enzyme has a relatively narrow substrate spectrum, preferably towards phosphotyrosine, phosphoserine and phosphoenolpyruvate. Interestingly, this enzyme was found to have a pH optimum at 8.5, rather than an acidic optima exhibited by eukaryotic PAPs. BcPAP contains a dimetallic ion centre composed of Fe and Zn, and site-directed mutagenesis confirmed that BcPAP utilizes the invariant residues for metal-ligation and catalysis. The enzyme is secreted by the wild type bacteria and its expression is regulated by the availability of orthophosphate. Our findings suggest that not all members in the PAP family have acidic pH optimum and broad substrate specificity.     

Protein Uptake and Digestion in Bloodstream and Culture Forms of Trypanosoma brucei*

SYNOPSIS. The mechanisms of ferritin uptake and digestion differ in bloodstream and culture forms of Trypanosoma brucei. Ferritin enters bloodstream forms from the flagellar pocket by pinocytosis in large spiny-coated vesicles. These vesicles become continuous with straight tubular extensions of a complex, mostly tubular, collecting membrane system where ferritin is concentrated. From the collecting membrane system the tracer enters large digestive vacuoles. Small spiny-coated vesicles, which never contain ferritin, are found in the Golgi region, fusing with the collecting membrane system, and around the flagellar pocket. Acid phosphatase activity is present in some small spiny-coated vesicles which may represent primary lysosomes. This enzymic activity is also found in the flagellar pocket, pinocytotic vesicles, the collecting membrane system, the Golgi (mature face), and digestive vacuoles of bloodstream forms. About 50% of the acid phosphatase activity of blood forms is latent. The remaining nonlatent activity is firmly cell-associated and probably represents activity in the flagellar pocket. The structures involved in ferritin uptake and digestion are larger and more active in the short stumpy than in the long slender bloodstream forms. The short stumpy forms also have more autophagic vacuoles. No pinocytotic large, spiny-coated vesicles or Golgi-derived, small spiny-coated vesicles are seen in culture forms. Ferritin leaves the flagellar pocket of these forms and enters small smooth cisternae located just beneath bulges in the pocket membrane. The tracer then passes through a cisternal collecting membrane network, where it is concentrated, and then into multivesicular bodies. In the culture forms, acid phosphatase activity is localized in the cisternal system, multivesicular bodies, the Golgi (mature face), and small vesicles in the Golgi and cisternal regions. The flagellar pocket has no acid phosphatase activity, and almost all the activity is latent in these forms. The culture forms do not release acid phosphatase into culture medium during 4 days growth. Uptake of ferritin by all forms is almost completely inhibited by low temperature. These differences among the long slender and short stumpy bloodstream forms and culture forms are undoubtedly adaptive and reflect different needs of the parasite in different life cycle stages.     

硫对土壤重金属形态转化及植物有效性的影响研究进展

生源要素硫在土壤中的化学循环不仅会直接影响土壤重金属元素的环境行为,也可通过调控植物根际微环境间接影响植物对重金属元素的吸收累积.土壤中的硫被植物根吸收后在植株中合成的有机硫化合物如植物螯合素(PCs)和金属硫蛋白(MTs)可与重金属形成毒性较低的络合物,构成植物重金属解毒的重要机制之一.我国部分土壤缺硫现象严重,为保证作物高质高产,硫肥的使用逐渐被重视,而硫与重金属的交互作用机制也逐渐成为研究热点.本文综合相关研究,介绍了硫在土壤中的生物化学转化,探讨了土壤硫的化学转化对土壤重金属形态转化及植物有效性的影响,并对今后硫在土壤重金属控制的应用提出展望.
     

The Fine Structure of the Centriolar Apparatus and Associated Structures in the Flagellates Deltotrichonympha and Koruga. I. Interphase

SYNOPSIS. An electronmicroscopic study was made of the centriolar apparatus in the rostrum of Deltotrichonympha operculata and Koruga bonita , 2 closely related hypermastigote flagellates from the Australian termite, Mastotermes darwiniensis. In interphase flagellates, the centriolar apparatus consists of 2 similar parts with a mutually perpendicular orientation. Each part contains a large, club-shaped centriolar body consisting of fibrillar and granular material, without recognizable internal symmetry or microtubules. The anterior centriolar body extends from the inner rostral wall, which is structurally related to the fibrous wall surrounding the posterior centriolar body. The 2 centriolar bodies are joined by connecting branches, which meet at 3 barren kinetosome-like structures located inside the rostrum. Thus, an interphase flagellate has 2 centriolar bodies oriented at a 90° angle to each other, like a pair of typical centrioles in an interphase metazoan cell.     

Pollen morphology of Xanthium L. (s.l.) (Asteraceae,Asteroideae, Heliantheae,Ambrosiinae) in the Iberian Peninsula - a palynotaxonomic approach to a poisonous,allergenic and invasive genus

Abstract

The pollen morphology of the Iberian taxa of the genus Xanthium s.l. and of the non-Iberian species X. ambrosioides was studied to help to identify these poisonous, allergenic and invasive taxa and clarify their taxonomic relationships. In addition, the question of the existence and structure of a mesoaperture in the apertural system of Xanthium was also considered. The pollen grains of sections Acanthoxanthium and Xanthium clearly differ by their characters, which supports the separation of both taxa and allows for their distinction based on pollen morphology. It is suggested that the morphological and palynological differences between Xanthium and Acanthoxanthium, together with the lack of hybridisation between the two sections and the monophyly of Acanthoxanthium, could lead to the elevation of the latter to the generic status. The two species of section Xanthium present in the Iberian Peninsula (X. orientale and X. strumarium) and of section Acanthoxanthium are well separated by their palynological features. Likewise, the Iberian subspecies of Xanthium, X. orientale subsp. orientale and X. orientale subsp. italicum are palynologically distinguishable. However, no significant difference could be found between the pollen of X. strumarium subsp. strumarium and of X. strumarium subsp. brasilicum. The pollen data do not support the hypotheses of a hybrid origin for X. strumarium subsp. brasilicum. A mesoaperture intersecting the foot layer and the upper part of the endexine is present in both sections, which is congruent with the results already found for other Asteroideae.     

赣江上游沉积物重金属空间分布及污染特征

为了解赣江上游表层沉积物重金属污染特征,采集赣江上游38个样品,采用电感耦合等离子体质谱仪(ICP-MS,Agilent 8800)分析沉积物中重金属含量,结合内梅罗综合污染指数法、地累积指数法、主成分分析及沉积物质量基准等研究赣江上游沉积物中重金属污染程度,分析污染物可能的来源及评价其存在的生态风险。结果表明:赣江上游沉积物中W、Cr、Mn、Cu、Zn、As、Cd、Hg、Pb等9种重金属的平均含量分别为12.30、4.40、999.84、9.72、293.81、47.66、2.85、1.34、63.68 mg·kg-1;除Cr和Cu外,其余7种重金属的平均值均超过江西省土壤背景值。内梅罗综合污染评价表明,赣江上游表层沉积物中47.37%的采样点呈严重污染,28.95%采样点污染明显,其中污染程度章水段>桃江段>贡江段;地积累指数法显示,Mn、As、W、Pb呈轻度污染,Cd和Hg呈中度或偏重度污染;主成分分析显示,As、W、Hg、Pb具有相似的污染源,Cr和Mn具有相似的污染源;沉积物质量基准分析表明,对底栖生物可能产生毒性效应的重金属主要是Cd和Hg,主要分布在章水段和桃...