4种蚯蚓肠道微生物对砷毒性的响应差异研究

蚯蚓肠道是微生物多样性的一个潜在存储库。砷对蚯蚓肠道微生物群落的影响已被证实,但砷在不同蚯蚓肠道菌群中生物转化的差异仍不清楚。为了进一步阐述土壤中广泛存在的低浓度砷(浓度为5,15,25 mg/kg)对不同种类蚯蚓肠道微生物影响的差异,将4种典型蚯蚓暴露于砷污染土壤后,测定其肠道微生物组成变化,并分析砷对不同蚯蚓肠道内砷富集、形态和砷生物转化基因的影响。结果显示,所有蚯蚓组织内均存在明显的砷富集,其富集系数由高到低依次为:安德爱胜蚓(1.93)>加州腔蚓(0.80)>通俗腔蚓(0.78)>湖北远盲蚓(0.52),蚯蚓组织和肠道内砷形态主要以无机砷为主,其中As(III)含量比例> 80%,部分蚯蚓组织内还发现少量有机砷。4种蚯蚓肠道微生物群落在门水平上主要以变形菌、厚壁菌和放线菌为主,并与周围土壤细菌群落组成存在显著差异。同时,在土壤和肠道内共检测到17个砷转化基因,其中蚯蚓肠道内As(V)还原和砷转运相关基因相对丰度较高,而砷(去)甲基化基因丰度较低。此外,低浓度砷污染对蚯蚓生长无显著影响,却能引起蚯蚓肠道微生物群落的紊乱。蚯蚓种类和砷污染是引起蚯蚓肠道微生物...     

Enantioselective Acute Toxicity Effects and Bioaccumulation of Furalaxyl in the Earthworm (Eisenia foetida)

The enantioselectivities of individual enantiomers of furalaxyl in acute toxicity and bioaccumulation in the earthworm (Eisenia foetida) were studied. The acute toxicity was tested by filter paper contact test. After 48 h of exposure, the calculated LC₅₀ values of the R‐form, rac‐form, and S‐form were 2.27, 2.08, and 1.22 µg cm^‐2, respectively. After 72 h of exposure, the calculated LC₅₀ values were 1.90, 1.54, and 1.00 µg cm^‐2, respectively. Therefore, the acute toxicity of furalaxyl enantiomers was enantioselective. During the bioaccumulation experiment, the enantiomer fraction of furalaxyl in earthworm tissue was observed to deviate from 0.50 and maintained a range of 0.55–0.60; in other words, the bioaccumulation of furalaxyl was enantioselective in earthworm tissue with a preferential accumulation of S‐furalaxyl. The uptake kinetic of furalaxyl enantiomers fitted the first‐order kinetics well and the calculated kinetic parameters were consistent with the low accumulation efficiency. Chirality 26:307–312, 2014. © 2014 Wiley Periodicals, Inc.     

The effect of anthropogenic arsenic contamination on the earthworm microbiome

Earthworms are globally distributed and perform essential roles for soil health and microbial structure. We have investigated the effect of an anthropogenic contamination gradient on the bacterial community of the keystone ecological species Lumbricus rubellus through utilizing 16S rRNA pyrosequencing for the first time to establish the microbiome of the host and surrounding soil. The earthworm‐associated microbiome differs from the surrounding environment which appears to be a result of both filtering and stimulation likely linked to the altered environment associated with the gut micro‐habitat (neutral pH, anoxia and increased carbon substrates). We identified a core earthworm community comprising Proteobacteria (～50%) and Actinobacteria (～30%), with lower abundances of Bacteroidetes (～6%) and Acidobacteria (～3%). In addition to the known earthworm symbiont (Verminephrobacter sp.), we identified a potential host‐associated Gammaproteobacteria species (Serratia sp.) that was absent from soil yet observed in most earthworms. Although a distinct bacterial community defines these earthworms, clear family‐ and species‐level modification were observed along an arsenic and iron contamination gradient. Several taxa observed in uncontaminated control microbiomes are suppressed by metal/metalloid field exposure, including eradication of the hereto ubiquitously associated Verminephrobacter symbiont, which raises implications to its functional role in the earthworm microbiome.     

Fungal volatilization of trivalent and pentavalent arsenic under laboratory conditions

Production of volatile derivatives of arsenic was studied using pure cultures of different fungal strains under laboratory conditions. Arsenic was used in its trivalent and pentavalent forms to evaluate the effect of arsenic valency on its biovolatilization. The average amount of volatilized arsenic for all fungal strains ranged from 0.026 mg to 0.257 mg and 0.024 mg to 0.191 mg of trivalent and pentavalent arsenic, respectively. These results show that approximately 23% of arsenic was volatilized from all culture media originally enriched with approximately 4 and 17 mg L(-1) of arsenic in trivalent form. The average amount of biovolatilized arsenic from culture media originally enriched with 4 and 17 mg L(-1) of arsenic in pentavalent form was 24% and 16%, respectively. The order of ability of arsenic biovolatilization is Neosartorya fischeri > Aspergillus clavatus > Aspergillus niger. Toxicity and fungal resistance to trivalent and pentavalent arsenic were also evaluated based on radial growth and biomass weight.     

Effect of arsenic trioxide on metallothionein and its conversion to different arsenic metabolites in hen liver

The metabolism of arsenic, its affinity to metallothionein (MT), its influence on selenium levels, and its biotransformation to different metabolites in the liver tissue of laying hens exposed to arsenic trioxide (As2O3) was investigated. The experiment was performed with two groups of hens fed for 19 d with either a standard diet or with the same diet enriched in arsenic (30 microg/g). The major findings were as follows: 1. After 19 d exposure, about 65% of the total liver As was found in the water-soluble phase (100,000g centrifuged supernatant). In liver supernatant, As binding was found mostly in the range of very low-molecular-weight proteins (Mr < 10,000). Although after exposure the amount of MT-like proteins increased, the As bound to it was only in trace amounts. The protein was identified by convential procedures as Zn,Cu-thionein with traces of selenium and arsenic. 2. Arsenic exposure resulted in almost unchanged Se levels regarding its tissue concentrations and distribution between supernatant and pellet, where about 10% of total Se was found in the supernatant. On the contrary, As exposure did affect Cd levels. Tissue Cd concentration was slightly diminished, but the percentage of tissue Cd found in the water-soluble phase was increased from 20% to 40%. 3. In methanol extracts of tissue and supernatant of the As-exposed group, only two arsenic compounds were detected, As(III) and dimethylarsinic acid (DMA), the latter prevailing.     

Prevention of arsenic-induced hepatic apoptosis by concomitant administration of garlic extracts in mice

Garlic is well known as a folk remedy for a variety of ailments since ancient times, however, very few studies are available suggesting its beneficial role against arsenic toxicity pertaining to its ability to eliminate arsenic from the blood and soft tissues and in reversal of arsenic-induced oxidative stress in affected tissues. The present study was planned to investigate the protective efficacy of aqueous garlic extract using two different doses on parameters suggestive of hepatic injury, tissue oxidative stress and mobilization of arsenic. Further, an attempt to understand the mechanism of arsenic in inducing hepatic apoptosis was also studied. Results of the present study suggested that arsenic administration in mice caused generation of reactive oxygen species (ROS) causing apoptosis through mitochondria-mediated pathway. The ROS generation in hepatic tissue reverted to normal values after co-administration of garlic extracts. The study provides significant evidence that garlic extracts contain strong anti-oxidant property which could be beneficial in preventing arsenic-induced toxicity in cells. However, further research is required to determine whether the results from animal studies are applicable to humans before garlic can be recommended as a putative agent against arsenic toxicity.     

Isolation from earthworms of a proteinaceous chemoattractant to garter snakes

We have isolated a potent proteinaceous chemoattractant from aqueous washes of earthworm (Lumbricus terrestris) for garter snakes (Thamnophis sirtalis) by means of a covalent chromatography. It contained free sulfhydryl groups and showed an apparent mass of 20 kDa. The chemoattractive activity of this protein could be destroyed by heating as well as by proteolysis. Its activity could also be reversibly blocked by mixed disulfide formation with dithiodipyridine, suggesting that the free sulfhydryl(s) was essential for its function as a chemoattractant. This bioactive material had a tendency to form intermolecular crosslinked aggregates during isolation, if reducing agents were not included. Some of the high-molecular-weight aggregates cochromatographed with earthworm cuticle collagen on Ultragel AcA 34 or 44 columns. In contrast to an earlier report by D. M. Kirschenbaum, N. Schulman, and M. Halpern [1986) Proc. Natl. Acad. Sci. USA 83, 1213-1216) the purified earthworm collagen showed no chemoattractive activity to garter snakes.     

The control of histone methylation and gene expression by oxidative stress, hypoxia, and metals

The harmful consequences of carcinogenic metals, such as nickel, arsenic, and chromium, are thought to be in part due to their ability to induce oxidative stress. The ubiquity of oxidative stress in biological systems has made it a fairly obvious culprit in causing cellular damage and/or development of disease. However, the full extent of oxidative stress-induced damage is not limited to its direct effects on cellular components, such as lipids, proteins, and DNA, but may extend to its ability to alter gene expression. Gene expression regulation is an important component of cellular and/or tissue homeostasis, and its alteration can have detrimental consequences. Therefore, a growing amount of interest is being paid to understanding how oxidative stress can influence gene expression. Oxidative stress-induced epigenetic dysregulation in the form of posttranslational histone modifications, in particular, is a popular topic of research. This review will therefore primarily focus on discussing the role of oxidative stress and hypoxia on histone methylation and/or gene expression alterations. The sources of oxidative stress discussed here are carcinogenic metals, such as, nickel, arsenic, and chromium.     

Mortality of the earthworm Eisenia fetida (Savigny) presented to the terrestrial planarian Ártioposthia triangulata (Dendy) (Tricladida: Terricola)

Ártioposthia triangulata is a terrestrial planarian that feeds on earthworms. Up to 76% of field collected A. triangulata fed on Eisenia fetida earthworms at 10°C in the laboratory. In one experiment, the feeding rate declined from 63% of earthworms attacked to 36% and 34% in the first, second and third feeding opportunities over a week. The average number of E. fetida eaten was 1.4 per planarian per week. The gain in weight of individual A. triangulata was significantly related to the amount of earthworm tissue lost during the feeding process. Over a one week period, the amount of earthworm tissue lost was related to the total amount presented to the planarians but not to their size. Frequency of attack was, however, related to planarian size. The results are discussed in relation to an hypothetical population of earthworms in pasture. It is concluded that the results support the contention that A. triangulata could seriously deplete earthworm populations.     

Heterologous expression of the yeast arsenite efflux system ACR3 improves Arabidopsis thaliana tolerance to arsenic stress

? Arsenic contamination has a negative impact on crop cultivation and on human health. As yet, no proteins have been identified in plants that mediate the extrusion of arsenic. Here, we heterologously expressed the yeast (Saccharomyces cerevisiae) arsenite efflux transporter ACR3 into Arabidopsis to evaluate how this affects plant tolerance and tissue arsenic contents. ? ACR3 was cloned from yeast and transformed into wild-type and nip7;1 Arabidopsis. Arsenic tolerance was determined at the cellular level using vitality stains in protoplasts, in intact seedlings grown on agar plates and in mature plants grown hydroponically. Arsenic efflux was measured from protoplasts and from intact plants, and arsenic levels were measured in roots and shoots of plants exposed to arsenate. ? At the cellular level, all transgenic lines showed increased tolerance to arsenite and arsenate and a greater capacity for arsenate efflux. With intact plants, three of four stably transformed lines showed improved growth, whereas only transgenic lines in the wild-type background showed increased efflux of arsenite into the external medium. The presence of ACR3 hardly affected tissue arsenic levels, but increased arsenic translocation to the shoot. ? Heterologous expression of yeast ACR3 endows plants with greater arsenic resistance, but does not lower significantly arsenic tissue levels.     

Protection of arsenic-induced testicular oxidative stress by arjunolic acid

Arsenic-induced tissue damage is a major concern to the human population. An impaired antioxidant defense mechanism followed by oxidative stress is the major cause of arsenic-induced toxicity, which can lead to reproductive failure. The present study was carried out to investigate the preventive role of arjunolic acid, a triterpenoid saponin isolated from the bark of Terminalia arjuna, against arsenic-induced testicular damage in mice. Administration of arsenic (in the form of sodium arsenite, NaAsO(2), at a dose of 10 mg/kg body weight) for 2 days significantly decreased the intracellular antioxidant power, the activities of the antioxidant enzymes, as well as the levels of cellular metabolites. In addition, arsenic intoxication enhanced testicular arsenic content, lipid peroxidation, protein carbonylation and the level of glutathione disulfide (GSSG). Exposure to arsenic also caused significant degeneration of the seminiferous tubules with necrosis and defoliation of spermatocytes. Pretreatment with arjunolic acid at a dose of 20 mg/kg body weight for 4 days could prevent the arsenic-induced testicular oxidative stress and injury to the histological structures of the testes. Arjunolic acid had free radical scavenging activity in a cell-free system and antioxidant power in vivo. In summary, the results suggest that the chemopreventive role of arjunolic acid against arsenic-induced testicular toxicity may be due to its intrinsic antioxidant property.     

Methylene blue filling of the dorsal giant fibres of the earthworm by electrophoresis

Multisegmental methylene blue filling of the three dorsal giant fibres in the nervous system of the earthworm was obtained by the use of electrophoresis. There was no difference in aldehyde fixed and unfixed tissue; due to its larger diameter, the median giant fibre could be stained over longer distances than the lateral ones. Accumulations of the dye were detected in the median giant fibre in the dorsal openings of the myelin sheath and at the origins of the ventral branches descending from both fibre systems. Since no significant axonal filling was observed by using several other dyes similar to methylene blue in diffusion and dialysis properties, the staining mechanism obviously depends on the specific binding sites of methylene blue in combination with the current. Therefore, the revealed phenomena provides evidence of a direct correlation between methyelne blue coupling and electrical coupling in the dorsal giant fibres of the earthworm.     

蚯蚓对细菌降解土壤中菲的作用

通过30d室内培养试验,分别研究了接种蚯蚓(E)、细菌(B)以及同时接种细菌和蚯蚓(BE)对土壤中菲降解的影响.结果表明: 在土壤中菲的初始污染浓度为50 mg*kg-1的条件下,各处理间菲的降解率差异显著,其降解率的大小顺序依次为:BE》B》E》CK(对照); 在150 mg*kg-1菲的初始污染浓度下,BE处理中菲的降解率高达98.86%,显著高于CK和E处理.B处理中细菌的双加氧酶活性在3种菲初始污染浓度下没有显著差异,而BE处理中双加氧酶的活性随着土壤中菲的初始污染浓度的升高而增加.在相同菲污染浓度下BE处理中蚯蚓体内的菲含量明显高于E处理.表明蚯蚓能够通过生物富集作用降低土壤中菲的浓度,而蚯蚓与细菌的相互作用能够进一步促进土壤中菲的降解.     

Molecular cloning and characterization of cDNA encoding fibrinolytic enzyme-3 from earthworm Eisenia foetida

Earthworm fibrinolytic enzyme (EFE), a multi-com-ponent protease purified from some earthworm breeds,belongs to serine protease family with fibrinolytic activity[1]. It has been used in prevention and treatment of cardiacand cerebrovascular diseases in Ch…     

Immobilization of arsenite and ferric iron by Acidithiobacillus ferrooxidans and its relevance to acid mine drainage

Weathering of the As-rich pyrite-rich tailings of the abandoned mining site of Carnoulès (southeastern France) results in the formation of acid waters heavily loaded with arsenic. Dissolved arsenic present in the seepage waters precipitates within a few meters from the bottom of the tailing dam in the presence of microorganisms. An Acidithiobacillus ferrooxidans strain, referred to as CC1, was isolated from the effluents. This strain was able to remove arsenic from a defined synthetic medium only when grown on ferrous iron. This A. ferrooxidans strain did not oxidize arsenite to arsenate directly or indirectly. Strain CC1 precipitated arsenic unexpectedly as arsenite but not arsenate, with ferric iron produced by its energy metabolism. Furthermore, arsenite was almost not found adsorbed on jarosite but associated with a poorly ordered schwertmannite. Arsenate is known to efficiently precipitate with ferric iron and sulfate in the form of more or less ordered schwertmannite, depending on the sulfur-to-arsenic ratio. Our data demonstrate that the coprecipitation of arsenite with schwertmannite also appears as a potential mechanism of arsenite removal in heavily contaminated acid waters. The removal of arsenite by coprecipitation with ferric iron appears to be a common property of the A. ferrooxidans species, as such a feature was observed with one private and three collection strains, one of which was the type strain.     

In vitro cultured cocklebur (Xanthium strumarium L.) responses to dimercaptopropanesulfonic acid and monosodium methanearsonate

Monosodium methanearsonate (MSMA) is an effective herbicide used for the control of Xanthium strumarium (cocklebur), a serious weed problem in cotton production. Recently, MSMA-resistant cocklebur was reported, which reduced MSMA effectiveness in the cocklebur control strategy. The mode of action of MSMA may involve the reduction of the pentavalent arsenic in MSMA to a trivalent form, which may then form a complex with sulfhydryl-containing enzymes. Dimercaptopro-panesulfonic acid (DMPS) chelates trivalent arsenic, resulting in a nontoxic complex. The objective of this study was to determine the effect of DMPS and MSMA on the growth of in vitro grown cocklebur shoot and callus tissues. Lateral and terminal shoots were grown on basal Murashige and Skoog medium containing 6.25 mg L^–1 (0.0428 mm) MSMA alone and with various concentrations of DMPS. Callus was cultured on basal callus medium also containing 6.25 mg L^–1 MSMA alone and an equimolar concentration of DMPS. Shoot injury symptoms, dry root weight, and fresh weight of callus were recorded. Susceptible cocklebur shoots showed severe discoloration and death when treated with MSMA. Resistant shoots and cultured shoot tips were not adversely affected. DMPS alone did not inhibit growth of these tissues. Callus induced from susceptible tissue was inhibited by MSMA and DMPS alone. In all combinations of DMPS and MSMA tested, DMPS did not counteract the MSMA response. Thus, the pentavalent form of arsenic in MSMA apparently is not reduced to the trivalent form by the plant and hence is not involved in the mechanism of actionAbbreviations MSMA monosodium methanearsonate - DMPS dimercaptopropanesulfonic acid - SM shoot medium - CM callus medium - ANOVA analysis of variance     

Distribution of arsenic in marine animals: relationship to diet

The chemical form of arsenic in the muscle tissues of marine animals in relation to diet has been examined. Inorganic arsenic concentrations in all muscle tissues were found to be low compared to total arsenic concentrations and independent of diet. Arsenic was found to be present in all tissues predominantly as a methanol-water soluble form. The ratio of lipid soluble and unextractable arsenic to total arsenic was higher in plankton feeders than in herbivores and carnivores.     

假单胞菌Tw224抗砷基因簇的功能研究

长期在砷污染环境生存的细菌逐渐演化出抗砷机制,旨在评价分离自湖南铅锌矿区污染土壤的Pseudomonas sp.Tw224抗砷能力和抗重金属谱;揭示该菌的抗砷基因/基因簇的功能.在砷或重金属存在条件下测量细菌的生长情况,对该菌的抗砷能力和抗重金属谱进行评价;通过基因组框架图分析该菌的抗砷基因/基因簇;采用代谢工程法构建...     

蚯蚓纤溶酶的纯化及稳定性研究

蚯蚓纤溶酶（ＥＰＡ）抽提液经３０％ ～７０％ 饱和度的（ＮＨ４）２ＳＯ４ 盐析、ＤＥＡＥ—纤维素柱层析、Ｓｅｐｈａｄｅｘ Ｇ－７５葡聚糖凝胶过滤等纯化步骤,得到了具有纤溶活性的洗脱峰,置凝胶电泳后,得到四个活性组分,它们经５０℃保温６ｈ,活力上升６４％ ;在２ ｍ ｏｌ／Ｌ盐酸胍存在时,活力仅保存７．２％ ,当其浓度降低时,活力可恢复至９０％ ;在１％ ＳＤＳ存在时,活力仅保存１２．１％ ,但当ＳＤＳ除去时,活力又可恢复。因此,盐酸胍、ＳＤＳ均为ＥＰＡ 可逆性抑制剂。另外,ＥＰＡ 中含有较高的糖链（占总量的４５％ ）,具有良好的抵抗自水解作用。