微生物砷还原机制的研究进展

砷是一种剧毒物质,环境中的砷对人体健康存在潜在威胁,因此长期以来备受关注.微生物的各种代谢过程对砷在环境中的归趋起着重要作用,其中砷还原微生物能将吸附于固体矿物中的As(Ⅴ)还原为可溶性强的As(Ⅲ),使砷进入液相,从而加剧了地下水等饮用水源的砷污染.论文主要介绍了两种微生物砷还原机制(异化砷还原和细胞质砷还原)在作用...     

抗氧化系统在蜈蚣草细胞砷解毒机制中的作用

砷超富集植物——蜈蚣草体内可以富集高浓度的砷且不表现出受害症状,暗示蜈蚣草细胞具有较强的砷解毒机制.为探讨抗氧化系统在蜈蚣草砷解毒机制中的作用,测定了0、0.5、1和2mmol/L的砷酸盐[As(Ⅴ)]、亚砷酸盐[As(Ⅲ)]和二甲基胂酸盐(DMA)处理下蜈蚣草愈伤组织的丙二醛(Malonyldiadehyde,MDA)含量、非酶类抗氧化物质含量和酶类抗氧化物质的活性.结果表明,0.5mmol/L As(Ⅴ)和DMA就可以诱导氧化胁迫的产生,浓度越高胁迫程度也越严重.而1mmol/L的As(III)仍未使蜈蚣草愈伤组织产生氧化胁迫.在As(Ⅲ)处理下,酸溶性巯基含量增加,叶绿素和类胡萝卜素含量不变,过氧化氢酶(Catalase,CAT)和过氧化物酶(Peroxidase,POD)活性增加.在As(Ⅴ)处理下,酸溶性巯基含量减少,叶绿素和类胡萝卜素含量略有上升;CAT和POD活性先增加后减少.在DMA处理下,酸溶性巯基含量减少,叶绿素含量降低而类胡萝卜素含量增加,CAT和POD活性先增加后减少.与As(Ⅴ)和DMA相比,As(Ⅲ)处理下高酸溶性巯基含量、高氧化酶活性和低MDA含量暗示As(Ⅲ)对蜈蚣草的毒性可能比As(Ⅴ)和DMA要小.这些结果表明,酸溶性巯基和酶类抗氧化物质(CAT和POD)在蜈蚣草细胞砷解毒机制中发挥着重要作用.图4参22     

绿藻胞外聚合物对无机砷生物累积特征的影响

绿藻对无机污染物的净化作用受其自分泌胞外聚合物EPS影响.以EPS释放量高的蛋白核小球藻为绿藻代表,通过24 h短期As(Ⅲ)和As(V)的模拟水体暴露实验,研究绿藻对无机砷的生物累积特征及EPS影响.结果表明,在0—40 mg·L~(-1) As(Ⅲ)和As(V)暴露浓度范围,蛋白核小球藻细胞内的砷累积速率随暴露浓度的增加而升高,其动力学拟合结果符合Michaelis-Menten酶促反应动力学方程. EPS与无机砷存在界面相互作用影响,无机砷暴露浓度升高可促进小球藻EPS分泌, EPS与砷累积速率之间呈现正相关线性关系(R~2 0.900),主要影响成分是溶解态EPS.完整藻细胞与脱除胞外聚合物的活体细胞相比, As(Ⅲ)、As(V)暴露的最大吸附累积量分别增加30.6%和14.2%,而最大胞内累积量降低49.0%和31.0%. EPS与无机砷的微界面交互作用影响绿藻对砷污染的净化修复.     

链霉菌的抗砷特性及其对蜈蚣草富集砷的作用

本文研究了链霉菌Streptomyces sp.的耐砷特性及其对蜈蚣草富集砷的影响。结果表明,Streptomyces sp.可在100mmo·lL-1的砷酸盐溶液中生长,具有较强的抗砷毒害能力,且在48h内对As(Ⅴ)的还原率达96.5%。施用Streptomyces sp.能促进植物对砷的吸收,蜈蚣草地上部砷浓度为930mg·kg-1,地上部砷累积量达到对照组的2.09倍。加入Streptomyces sp.后,能促进根际土壤中As(Ⅴ)还原成As(Ⅲ),大幅度降低根际土壤残渣态砷含量,从48.15mg·kg-1下降至28.75mg·kg-1。Streptomyces sp.通过影响蜈蚣草根际环境,提高根际土壤pH,增加DOC含量,促使砷形态变化,从而增加砷生物可利用性。该菌可作为强化蜈蚣草修复砷污染土壤的材料。     

氢化物原子荧光光谱法测定水中痕量砷（Ⅲ）和砷（Ⅴ）

应用氢化物发生无色散原子荧光法测定水中痕量As(Ⅲ)和As(Ⅴ).在pH5.6—6.0时,As(Ⅲ)与KBH_4作用生成气态氢化物(AsH_3),被原子荧光仪测定.在此酸度下,AS(Ⅴ)不发生反应.在2NHCl溶液中,用硫脲和抗坏血酸还原As(Ⅴ)为As(Ⅲ),同法测总砷,用差减法求得As(Ⅴ).方法检出限0.1ppb,相对标准偏差4.6—5.8％,回收率93—104％.     

湖南桃江锰矿对溶液中As(Ⅴ)和As(Ⅲ)的去除及迁移行为对比

锰氧化物对砷的去除有着环境和地球化学现实意义,本文通过批实验和柱实验研究湖南桃江锰矿对溶液中As(Ⅴ/Ⅲ)的去除行为差异和迁移行为.Langmuir吸附等温线结果表明,锰矿对As(V)和As(Ⅲ)的理论最大吸附量分别为1.32 mg·g~(-1)和0.30 mg·g~(-1).As(Ⅴ/Ⅲ)在锰矿表面的动力学符合拟二阶动力学模型,表明锰矿吸附As(Ⅴ/Ⅲ)均属于化学吸附,受化学反应速率控制;反应吸附速率常数K_(2[As(Ⅴ)])K_(2[As(Ⅲ)]),表明锰矿对As(Ⅴ)的吸附速率更快.锰矿在氧化As(Ⅲ)时,溶液中As(Ⅲ)减少速率与溶液中Mn浓度变化非常一致,表明砷的氧化行为与锰矿相关.CDE和Thomas吸附模型拟合As(Ⅴ)和As(Ⅲ)的迁移行为表明,Mn~(2+)、Al~(3+)、PO■和SiO■均会降低锰矿的最大吸附量和滞留因子,其中Al~(3+)对As(Ⅲ)的吸附具有较强的拮抗作用,最大吸附量下降至0.002 mg·g~(-1),对As(Ⅴ)的拮抗作用相对较弱,而PO■和SiO■对As(Ⅴ/Ⅲ)在柱实验中的拮抗作用相近.研究为就地处理湖南地表和地下水砷污染提供了新的处理矿物.     

锑和砷对固氮菌的毒性效应及其机制研究

锑作为一种具有毒性和致癌性的类金属,其不合理的开发造成了严重的生态环境污染,尤其是土壤中高浓度锑和砷污染导致了环境中多种营养元素的缺乏。其中,氮素的缺乏严重阻碍了环境的恢复。然而,固氮微生物不仅能为氮素匮乏环境提供稳定的氮素来源,而且其固氮潜能可以作为监测土壤金属污染的敏感指标。为此,该文通过乙炔还原法(ARA)微宇宙培养试验手段,研究了不同价态锑和砷(三价及五价)在不同质量浓度(100、200、500、1 000、2 500、5 000 mg·L^-1)水平下对棕色固氮菌固氮潜能的影响,同时进一步探究了土壤固氮菌群落固氮潜能对土壤中主要锑和砷价态[Sb(Ⅴ)和As(Ⅴ)]的响应机制。结果表明,锑和砷浓度与棕色固氮菌和土壤固氮微生物的固氮潜能均呈显著负相关,验证了以固氮微生物固氮潜能作为生物指示物的可行性。此外,Sb(Ⅴ)对土壤固氮微生物的毒性显著小于As(Ⅴ)。三价锑和砷污染对固氮菌固氮潜能的毒性高于五价[As(Ⅲ)>Sb(Ⅲ)>As(Ⅴ)>Sb(Ⅴ)],这可能是由于Sb(Ⅲ)和As(Ⅲ)对细胞的致裂性强于Sb(Ⅴ)和As(Ⅴ)。相较于Sb(Ⅴ)和As(Ⅴ)对于棕色固氮菌固氮潜能的抑制作用,其对土壤固氮菌群落的固氮潜能的影响相对较弱。这一差异可能是由于土壤复杂的理化性质以及微生物群落和功能多样性而导致的。     

利用高效液相色谱-动态反应池模式-等离子体质谱法测定水稻中的砷形态

利用HPLC-DRC-ICP-MS测定了水稻中4种砷的形态As(Ⅲ)、DMA(Ⅴ)、MMA(Ⅴ)、As(Ⅴ),比较了两种提取剂的优劣.结果表明,三氟乙酸提取剂在提取率及加标回收率两方面均优于甲醇-水.HPLC-DRC-ICP-MS具快速、可自动分析的特点,提取液中上述4种不同形态的砷可在7min内采用等度洗脱法得以有效地分离,其检测限低于0.1 mg·kg-1.     

植物砷吸收与代谢的研究进展

砷(As)作为一种植物非必需的类金属元素广泛存在于自然界中,砷过量摄人不仅会对植物生长产生毒害作用,而且在植物的可食部位累积并通过食物链对人体健康构成威胁.生长介质中的砷酸盐(五价砷)一般是通过磷酸盐转运蛋白被植物吸收的,而亚砷酸(三价砷)和没有解离的甲基化砷则主要是通过质膜上的水通道蛋白被植物吸收的.在植物体内五价砷...     

微生物砷代谢机制的研究进展（Progress in Study of Mechanisms of Microbial Arsenic Transformation in Environment）

环境砷污染是一个全球性问题.研究砷的生物地球化学循环可以明确环境中砷的来源及其转化特征,为探索砷污染治理的方法提供参考.越来越多的研究表明,自然界中的微生物在砷的迁移转化过程中发挥了重要作用.根据微生物对砷的代谢机制不同将其分为:砷氧化微生物、砷还原微生物和砷甲基化微生物.砷氧化微生物可以将环境中的As(Ⅲ)氧化为毒性较弱并且容易被铁铝矿物吸附固定的As(Ⅴ),因此对降低环境中的砷毒性具有重要作用;微生物对砷的甲基化作用的产物通常为毒性较低的有机砷,因此也被认为是理想的修复环境砷污染的生物手段之一;然而在还原环境中,砷还原微生物却可以将游离态和结合态的As(Ⅴ)还原为毒性更强的As(Ⅲ),从而加重环境中的砷污染状况.由此可见,明确微生物的砷代谢机制及其对砷污染环境中砷迁移转化的影响,是实现生物修复砷污染环境的必要前提.论文总结了近年来国内外微生物砷代谢机制的研究进展,以期为深入研究微生物代谢砷的机理及其在砷污染治理中的应用提供参考.     

Non-carcinogenic effects of inorganic arsenic

This review will focus primarily on ohe effects of the inorganic arsenicals (arsenate and arsenite forms) that are present in drinking water. They are acutely toxic to both humans and animals, an effect that may be related to their bioavailibility. In humans, arsenicals have been reported to cause dermatitis and mucous membrane irritation upon exposure. They have also been reported to cause skin lesions and peripheral neurotoxicity in smelter workers and in patients treated with Fowler's Solution. When humans are exposed to arsenic in drinking water, effects such as hyperkeratosis, electromyographic abnormalities and vascular effects have been reported. In experimental animals, arsenic has been demonstrated to affect the liver and kidneys. In mice, arsenic has also been reported to decrease the animal's resistance to certain viral infections. The arsenite (+3) and arsenate (+5) forms have different modes of action. Arsenite binds to sulphhydryl groups and has been reported to inhibit over 100 different enzymes, while the arsenate can substitute for phosphate in various high energy intermediates, resulting in arsenolysis. In addition, when arsenate is reduced to arsenite in the body, it can also cause toxicity as that species.     

Characteristics of arsenic accumulation by the seaweeds Fucus spiralis and Ascophyllum nodosum

This study examines the accumulation of ⁷⁴As-arsenic in the seaweeds Fucus spiralis (L.) and Ascophyllum nodosum (L.) collected from Restronguet Creek in southwest England during 1978. Also, the influence of environmental factors (valence state of arsenic, pH, salinity, temperature, phosphate concentration) and metabolic inhibitors on the uptake of arsenic by F. spiralis is examined. Most of the arsenic in the seaweeds was non-exchangeable with labelled arsenic in the medium. The accumulation of ⁷⁴As reached a steady state in 1 to 8 d, depending on the species and external arsenic concentration. At steady state the accumulated arsenic is proportional to external arsenate concentration. F. spiralis accumulated 4 times more arsenate than arsenite. The short-term uptake of arsenic increased in proportion to the external arsenic concentration up to a level of 1000 g l^-1; it then remained constant at higher levels of arsenic. Arsenic uptake increased in direct proportion to increasing temperature. Variation of pH or salinity had no effect on arsenic incorporation. The accumulation of arsenic occurred only in living tissue and was inhibited by KCN in a concentration-dependent manner. The uptake of arsenic by F. spiralis in the presence of photo-synthetic inhibitors (DCMU or CMU) or in the dark was greater than in the light controls. Thus, it was concluded that energy is required for arsenic uptake and this is derived from respiration rather than photosynthesis. There was no evidence for a common mechanism of phosphate and arsenate uptake by macroalgae, although high concentrations of phosphate (40 to 400 M) initially inhibited arsenate uptake.     

Uptake of arsenate, trimethylarsine oxide, and arsenobetaine by the shrimp Crangon crangon

Common shrimp, Crangon crangon (L.), were exposed to inorganic arsenic (arsenate), trimethylarsine oxide, or arsenobetaine in sea water (100 μg As l⁻¹) or in food (1 mg As g⁻¹ wet wt) for up to 24 d, followed by 16 d depuration in clean sea water with undosed food, in order to determine the efficiency of uptake and retention of the compounds. Accumulation of arsenic in the tail muscle, gills, midgut gland, exoskeleton, and remaining tissues was found to depend on the chemical form of the arsenic and the route of exposure. No arsenic was accumulated by C. crangon exposed to arsenate or trimethylarsine oxide in sea water. Shrimps exposed to waterborne arsenobetaine initially accumulated a small amount of arsenic in their tail muscle and gills. After 16 d, C. crangon fed arsenate, trimethylarsine oxide, or arsenobetaine had accumulated arsenic in their tail muscle to levels ∼2-, 2-, or 40-times, respectively, that of the control group. A roughly linear rate of accumulation was shown by shrimps fed trimethylarsine oxide or arsenobetaine, but C. crangon fed arsenate accumulated arsenic for 16 d, then lost arsenic such that their concentration on Day 24 was not significantly different from that of the control group. Patterns of arsenic accumulation in the gills of shrimps fed the compounds were similar to those seen in the tail muscle. On a whole animal basis, C. crangon retained ∼1.2% of the arsenate, 1.6% of the trimethylarsine oxide, and 42% of the arsenobetaine consumed over the first 16 d of exposure, with roughly half present in the tail muscle in each case. Data obtained support the view that the direct uptake of arsenobetaine from sea water does not make a significant contribution to the relatively high concentrations of this compound in marine crustaceans, and that food is the primary source. Naturally occurring arsenic compounds in C. crangon and possible transformations of the administered arsenic compounds were examined by high performance liquid chromatography using an inductively coupled plasma mass spectrometer as the arsenic-specific detector. Control C. crangon contained arsenobetaine as the major arsenic compound (>95% of total arsenic); tetramethylarsonium ion (0.7%) and an unknown arsenic compound (1.7%) were also present as minor constituents. Shrimp ingesting arsenobetaine accumulated it unchanged. Shrimp ingesting arsenate did not form methylated arsenic compounds; they appeared to contain their accumulated arsenic as unchanged arsenate only, although the possibility that some of the arsenic was reduced to arsenite could not be excluded. C. crangon ingesting trimethylarsine oxide biotransformed the compound predominantly to dimethylarsinate. Received: 9 October 1997 / Accepted: 11 February 1998     

Species-level study on arsenic availability from dietary components

Arsenic (As) contaminated water and foodstuffs are of major concern. Samples of drinking–cooking water (n = 50), raw rice (n = 50), common vegetables (eight types), and common pulses (three types) were collected from households in the endemic region. The study found up to 70% As reduction by using safe water for cooking of rice and vegetables. Speciation study reflected more arsenate than arsenite and other organic arsenicals in all the types of samples. Male intake of 293 μg As through drinking water contained 38 μg arsenite and 246 μg arsenate, and female intake of 199 μg As contained 167 μg arsenate and 25 μg arsenite. In cooked rice, 108 μg As contained 69 μg arsenate and 17 μg arsenite with 9 μg dimethylarsonic acid (DMA). Total As consumption from cooked vegetables was 45 μg with 34 and 4 μg of arsenite and arsenate, respectively, and 5 μg of DMA. Data indicate that cooking with As-free water removes arsenic in already contaminated foodstuffs but without interconversion of the As species, from toxic inorganic to less toxic organic forms.     

砷浓度、形态及碳酸氢盐对蜈蚣草吸收砷的影响

为了探讨超富集植物蜈蚣草在处理高砷地下水方面的可行性,研究了水培条件下砷的浓度、形态和碳酸氢盐(HCO-3)对超富集植物蜈蚣草吸收砷的影响。实验中使用了浓度为0.1～100mg·L-1的As(III)和As(V)溶液。HCO-3处理中,HCO-3浓度范围为0.5～20mmol·L-1,As(III)或As(V)的浓度为5mg·L-1。结果表明,在水培条件下,蜈蚣草具有明显的耐高砷特征。当介质砷含量高达100mg·L-1时,砷的去除率可达到80%,且对As(III)的吸收效率高于As(V)。植物体内砷形态研究表明,蜈蚣草体内2种形态砷的含量与外源砷形态有一定的关系,As(V)处理条件下,植物体中的As(V)比例较As(III)处理高。高浓度的HCO-3(20mmol·L-1)处理对蜈蚣草地上部分生物量没有明显影响,但是抑制了地下部分的生长,并且对砷的吸收表现出明显的抑制作用。     

砷浓度、形态及碳酸氢盐对蜈蚣草吸收砷的影响

大同盆地是典型的高砷地下水分布区。利用从地方性砷中毒严重病区山阴县采集的高砷地下水样品,用稀释培养法实验研究了外加砷源对地下水中微生物数量的影响;同时基于生物学可培养法和16S rDNA序列比对法,选取代表性高砷水样,研究了耐砷菌的种群特征。结果表明,外加砷源对地下水中微生物数量影响显著,高浓度砷会抑制大部分微生物生长,使微生物数量减少;低浓度砷对微生物生长具有一定促进作用。通过多次分离、纯化从3个不同砷含量地下水样中分离到多株砷抗性菌,经鉴定属于主要为Bacillus、Pseudomonas、Paenibacillus、Aeromonas、Enterobacter5个属。从RDP(Ribosomal Database Project)分析显示3个水样可培养微生物组成不同,都有生存能力强能够耐低浓度NaAsO₂的Bacillales,优势耐砷菌是γ-proteobacteria,其中Enterbacter具有耐高浓度NaAsO₂的能力。     

不同磷浓度下水稻短根突变体与野生型对五价砷吸收和转运的差异

为研究水稻根形态及磷营养状况对水稻吸收和转运五价砷的调控作用,采用水培的方法,研究了在不同外部磷浓度(0、10、50、150、300μM KH2PO4)下水稻(Oryza sativa L.)短根突变体与野生型短期内对五价砷(10μM Na3AsO4)吸收和转运的差异.结果表明,水稻根形态(短根突变体与野生型)和磷营养状况均能对水稻吸收和转运五价砷产生显著影响:1)磷能竞争性抑制水稻对五价砷的吸收,随营养液中磷浓度的增加,水稻地上部和地下部五价砷含量、单位根干重砷吸收量(根系砷吸收能力)均显著降低;2)水稻短根突变体对五价砷的吸收能力低于野生型,但转运能力高于野生型.     

砷形态在湖水中垂直分布的研究

本文以天津水上公园湖泊作为研究现场，通过采样分析，研究了湖水中砷酸盐、亚砷酸盐、一甲基胂酸盐和二甲基胂酸盐的垂直分布及其影响因素。结果表明水上公园湖泊中砷酸盐是溶解态砷的主要存在形式，其垂直分布与悬浮物、叶绿素ａ、磷酸盐的浓度、水文等诸因素密切相关；一甲基胂酸盐含量保持相对稳定；光致转化是影响水中亚砷酸盐垂直分布差异的一个因素。     

Arsenic metabolism in marine bacteria and yeast

Arsenic metabolism was studied for two marine microorganisms, a facultative anaerobic bacterium, Serratia marinorubra, and an obligately aerobic yeast, Rhodotorula rubra. Both were cultivated in media with (⁷⁴As) arsenate (As V), and the products of arsenate metabolism were determined qualitatively. Both the bacterium and the yeast produced arsenite (AS III) and methylarsonic acid [CH₃AsO(OH)₂]. In addition to the foregoing, only the yeast produced dimethylarsinic acid (CH₃)₂AsO(OH) and volatile alkylarsines. In contrast, the bacterium growing anaerobically with cobalamine as a cofactor did not synthesize gaseous forms of arsenic such as methylarsines. Neither organism synthesized arsoniumphospholipids such as those produced by marine phytoplankton or terrestrial fungi. The yeast did not accumulate arsenite, but instead transported some of it into the culture medium and methylated the remainder first to methylarsonic acid and then to dimethylarsinic acid. Finally, the latter compound was methylated further and volatile alkylarsines were formed. In contrast, the bacterium retained all products of arsenate metabolism intracellularly. Both the bacterium and the yeast, therefore, converted relatively toxic arsenate, the most abundant arsenic compound in seawater, to products that were presumably less toxic.