Production and release of selenocyanate by different green freshwater algae in environmental and laboratory samples

In a previous study, selenocyanate was tentatively identified as a biotransformation product when green algae were exposed to environmentally relevant concentrations of selenate. In this follow-up study, we confirm conclusively the presence of selenocyanate in Chlorella vulgaris culture medium by electrospray mass spectrometry, based on selenium's known isotopic pattern. We also demonstrate that the observed phenomenon extends to other green algae (Chlorella kesslerii and Scenedesmus obliquus) and at least one species of blue-green algae (Synechococcus leopoliensis). Further laboratory experiments show that selenocyanate production by algae is enhanced by addition of nitrate, which appears to serve as a source of cyanide produced in the algae. Ultimately, this biotransformation process was confirmed in field experiments where trace amounts of selenocyanate (0.215 ± 0.010 ppb) were observed in a eutrophic, selenium-impacted river with massive algal blooms, which consisted of filamentous green algae (Cladophora genus) and blue-green algae (Anabaena genus). Selenocyanate abundance was low despite elevated selenium concentrations, apparently due to suppression of selenate uptake by sulfate, and insufficient nitrogen concentrations. Finally, trace levels of several other unidentified selenium-containing compounds were observed in these river water samples; preliminary suggestions for their identities include thioselenate and small organic Se species.     

Characterization of Pseudomonas stutzeri NT-I capable of removing soluble selenium from the aqueous phase under aerobic conditions

Pseudomonas stutzeri strain NT-I was isolated from the drainage wastewater of a selenium refinery plant. This bacterium efficiently reduced selenate to elemental selenium without prolonged accumulation of selenite under aerobic conditions. Strain NT-I was able to reduce selenate completely at high concentrations (up to 10 mM) and selenite almost completely (up to 9 mM). In addition, higher concentrations of selenate and selenite were substantially reduced. Activity was observed under the following experimental conditions: 20–50°C, pH 7–9, and 0.05–20 g L^–1 NaCl for selenate reduction, and 20–50°C, pH 6–9, and 0.05–50 g L^–1 NaCl for selenite reduction. Under anaerobic conditions, selenate was reduced more rapidly, whereas selenite was not reduced at all. The high selenate- and selenite-reducing capability at high concentrations suggested that strain NT-I is suitable for the removal of selenium from high-strength industrial wastewater.     

Factors affecting soluble selenium removal by a selenate-reducing bacterium Bacillus sp. SF-1

High concentrations of soluble selenium, selenate and selenite, have acute and chronic toxicity toward living things. With the aim of developing a biological process for selenium removal, the effects of a variety of parameters on the reduction of soluble selenium by a Bacillus sp. strain SF-1, which is capable of reductively transforming selenate into selenite and, subsequently, into nontoxic insoluble elemental selenium, were studied. The bacterial strain could effectively reduce 20 mM of selenate to selenite and 2 mM of selenite to elemental selenium in the presence of an appropriate carbon source and in the absence of oxygen. The reduction rate of selenate to selenite was much higher than that of selenite to elemental selenium, resulting in the transient accumulation of selenite during selenate reduction. The selenate reduction rate increased with increases in the selenate concentration up to 20 mM, while the rate of selenite reduction decreased sharply at selenite concentrations of more than 2 mM. The elemental selenium transformed from selenate via selenite was found both inside and outside the cells. Bacillus sp. SF-1 was able to utilize a variety of organic acids or sugars as a carbon source in selenate reduction. Although the copresence of sulfate did not inhibit selenate reduction, it was completely inhibited by some other oxyanions, including nitrate. A model sequencing batch system using the bacterial strain was developed and exhibited good performance in the treatment of wastewater containing high concentrations of selenate.     

Factors affecting soluble selenium removal by a selenate-reducing bacterium Bacillus sp. SF-1

High concentrations of soluble selenium, selenate and selenite, have acute and chronic toxicity toward living things. With the aim of developing a biological process for selenium removal, the effects of a variety of parameters on the reduction of soluble selenium by a Bacillus sp. strain SF-1, which is capable of reductively transforming selenate into selenite and, subsequently, into nontoxic insoluble elemental selenium, were studied. The bacterial strain could effectively reduce 20 mM of selenate to selenite and 2 mM of selenite to elemental selenium in the presence of an appropriate carbon source and in the absence of oxygen. The reduction rate of selenate to selenite was much higher than that of selenite to elemental selenium, resulting in the transient accumulation of selenite during selenate reduction. The selenate reduction rate increased with increases in the selenate concentration up to 20 mM, while the rate of selenite reduction decreased sharply at selenite concentrations of more than 2 mM. The elemental selenium transformed from selenate via selenite was found both inside and outside the cells. Bacillus sp. SF-1 was able to utilize a variety of organic acids or sugars as a carbon source in selenate reduction. Although the copresence of sulfate did not inhibit selenate reduction, it was completely inhibited by some other oxyanions, including nitrate. A model sequencing batch system using the bacterial strain was developed and exhibited good performance in the treatment of wastewater containing high concentrations of selenate.     

烟株对不同价态硒的吸收与分配

为了开发富硒烟叶生产技术和了解烟叶对硒的富集机理,采用水溶性Se（Ⅳ）（亚硒酸钠）和Se（Ⅵ）（硒酸钠）盆栽试验,探讨了烟株对硒的吸收和分配规律,以及不同价态和用量的硒对烟株生长发育的影响。结果表明,土壤施用适当浓度的两种价态硒（1～10 mg/kg）均有改善烟株形态性状和提高生物量的作用,高浓度硒（20～30 mg/kg）则有负面影响,且Se（Ⅵ）处理抑制效果大于 Se（Ⅳ）。相同浓度下,两种价态的硒均能有效提高烟株各部位硒含量,Se（Ⅵ）处理较 Se（Ⅳ）提高了3～30倍。土壤中的硒优先富集和分布于叶片中,其次是根系,茎秆最小。Monod方程拟合结果表明,Se（Ⅵ）处理烟株叶片、茎秆和根系最大硒含量分别是Se（Ⅳ）处理的1.83、15.81和20.98倍,烟株对Se（Ⅵ）的亲和能力显著大于Se（Ⅳ）,说明Se（Ⅵ）对烟株的生物有效性明显高于Se（Ⅳ）。     

Combined speciation analysis by X-ray absorption near-edge structure spectroscopy, ion chromatography, and solid-phase microextraction gas chromatography-mass spectrometry to evaluate biotreatment of concentrated selenium wastewaters

In this study we evaluate the potential of anaerobic granular sludge as an inoculum for the bioremediation of selenium-contaminated waters using species-specific analytical methods. Solid species formed by microbial reduction were investigated using X-ray absorption near-edge structure (XANES) spectroscopy at the selenium K-edge. Furthermore, dissolved selenium species were specifically determined by ion chromatography (IC) and solid-phase microextraction gas chromatography-mass spectrometry (SPME-GC-MS). Least-squares linear combination of the XANES spectra for samples incubated with the highest selenate/selenite concentrations (10(-3) M) show the predominance of elemental selenium and a Se(-I) selenide, such as ferroselite, the thermodynamically most stable iron selenide. In contrast, elemental selenium and Se(-II) selenides are the main species detected at the lower selenate/selenite concentrations. In each repeated fed batch incubation, most aqueous selenite anions were converted into solid selenium species, regardless of the type of electron donor used (acetate or H(2)/CO(2)) and the selenium concentration applied. On the other hand, at higher concentrations of selenate (10(-4) and 10(-3) M), significant amounts of the oxyanion remained unconverted after consecutive incubations. SPME-GC-MS demonstrated selenium alkylation with both electron donors investigated, as dimethyl selenide (DMSe) and dimethyl diselenide (DMDSe). Selenite was even more alkylated in the presence of H(2)/CO(2) (maximum 2156 μg of Se/L of DMSe + DMDSe) as compared to acetate (maximum 50 μg of Se/L). In contrast, selenate was less alkylated using both electron donors (maximum 166 and 3 μg of Se/L, respectively). The high alkylation potential for selenite limits its bioremediation in selenium laden waters involving H(2)/CO(2) as the electron donor despite the fact that nontoxic elemental selenium and thermodynamically stable metal selenide species are formed.     

Identification and determination of selenosulfate and selenocyanate in flue gas desulfurization waters

In this work, 13 selenium species in flue gas desulfurization (FGD) waters from coal-fired power plants were separated and quantified using anion-exchange chromatography coupled to inductively coupled plasma mass spectrometry. For the first time, we identified both selenosulfate (SeSO(3)(2-)) and selenocyanate (SeCN(-)) in such waters, using retention time matching and confirmation by electrospray mass spectrometry. Besides selenite and selenate, selenosulfate was the most frequently occurring selenium species. It occurred in most samples and constituted a major fraction (up to 63%) of the total selenium concentration in waters obtained from plants employing inhibited oxidation scrubbers. Selenocyanate occurred in about half of the tested samples, but was only a minor species (up to 6% of the total selenium concentration). Nine additional Se-containing compounds were found in FGD waters, but they remain unidentified at this point.     

Response of sediment bacterial assemblages to selenate and acetate amendments

We followed the response of bacterial assemblages in slurries of estuarine sediments to amendments of 100 microM sodium selenate and 10 mM sodium acetate. Selenate was removed from the dissolved phase in all slurries after an initial lag that varied depending on the source of the sediment used in the slurry. Subsequent selenate amendments were removed without a lag but with the appearance of transient peaks of selenite. We documented changes in the composition of bacterial assemblages in the slurries using PCR/DGGE and RT-PCR/DGGE of 16S rDNA and rRNA. Bands of interest were sequenced to identify organisms responding to selenate amendments. The composition of communities from the two sampling sites was different and changed but did not converge during incubations with selenate. Selenate-reducing assemblages had broad phylogenetic diversity. Most bands were related to groups of bacteria known to contain organisms capable of selenate or selenite reduction, except for Acinetobacter species which dominated one of the samples and has not previously been associated with selenium oxyanion reduction.     

Uptake and distribution of added selenite and selenate by barley and red clover as influenced by sulphur

The uptake of added Se from selenite and selenate by barley and red clover was investigated in a pot experiment. Much more of selenate than of selenite was taken up but the Se concentrations in the plants declined more with time when selenate was the source. Increasing sulphate addition to the soil decreased the uptake of Se from selenate greatly and from selenite to a lesser extent. The ratio Se in the roots/Se in the tops shows that Se is more readily translocated from the roots when taken up from added selenate than from selenite. On the basis of these and other investigations it is concluded that selenite is a better source than selenate when the Se concentration in the plants has to be raised to a level sufficient for livestock nutrition.     

The effect of heat treatment on intrinsic and fortified selenium levels in cow's milk

The effects of two heat processing methods (pasteurisation and spray drying) routinely used in the processing of cow's milk and the production of infant formula powder on the selenium (Se) content of liquid milk, milk fortified with sodium selenite and sodium selenate were studied. Pasteurisation reduced intrinsic Se and selenate levels by 7.9% and 6.2% at p < 0.05 level and selenite levels by 7.0% at p > 0.05 level. Se losses following spray drying were 44.8% (p < 0.001), 11.4% (p < 0.01) and 10.0% (p < 0.01) for intrinsic selenium, selenite and selenate fortified milk, respectively. Total Se losses from unprocessed milk following processing (pasteurisation and spray drying) were 49.2% (p < 0.001), 17.6% (p < 0.001) and 15.6% (p < 0.001) for intrinsic selenium, selenite and selenate fortified milk, respectively.     

转基因大豆对硒的富集作用和形态分布研究

目的研究转基因大豆对硒的富集作用以及富集前后转基因大豆中硒的形态分布和形态转化。方法 采用电感耦合等离子体质谱分析转基因大豆富集前后硒的总量,采用蛋白酶提取,高效液相色谱-电感耦合等离子体质谱(high performance liquid chromatography-inductively coupled plasma-mass spectrometry,HPLC-ICP-MS)联用技术分析转基因大豆中的硒酸盐(selenate,Se VI)、亚硒酸盐(selenite,Se IV)、硒代蛋氨酸(selenomethionine,Se Met)和硒代胱氨酸(selenocystine,Se Cys)等几种硒化合物。盆栽种植试验研究转基因大豆对硒的富集作用。通过对照试验,考察大豆植株不同部位总硒含量变化和形态分布的转化情况。结果转基因大豆对硒有较好的富集作用,吸收的亚硒酸盐在大豆植株中部分转化成了硒代蛋氨酸和硒代胱氨酸。结论 了解了转基因大豆的形态分布和转化情况,可以更好地评估转基因大豆的食用安全风险并进行生物利用开发。     

硒酸化乳清分离蛋白抗前列腺癌细胞活性

采用干燥加热法制备硒酸化乳清分离蛋白(selenized whey protein isolate,Se-WPI),通过Se-WPI中有机硒质量分数、硒酸根稳定性、Se-WPI消化率测定,⁷⁷Se核磁共振(nuclear magnetic resonance,NMR)分析以及Se-WPI抑制前列腺癌细胞增殖实验等,研究其理化性质及抗前列腺癌细胞活性。结果表明,在pH 3.0、加热温度80℃、加热时间24 h条件下制备得到的Se-WPI中有机硒质量分数为2.09%。⁷⁷Se-NMR和硒酸根稳定性分析结果表明,Se-WPI中的硒以亚硒酸酯的形式存在。经硒酸化的WPI多种生物活性得到改善,消化性明显提高。Se-WPI与亚硒酸钠均具有抑制人前列腺癌LNCaP细胞、DU145细胞增殖的活性。细胞周期、细胞凋亡和Caspase-3活力检测结果从不同角度验证了Se-WPI具有诱导LNCaP细胞凋亡的作用。另外,Se-WPI还能抑制癌细胞与基底膜成分的黏附能力。实验结果可为进一步研究有机硒化合物的抑癌机制提供依据。     

耐硒芽孢杆菌的筛选及其亚硒酸盐还原机制的探究

微生物能将氧化形态的有毒硒转化为无毒的纳米硒（SeNPs）,这种“绿色合成”方式在功能性产品开发、环境治理和医疗等方面的应用潜力巨大。本文从长期施加硒肥的大豆种植基地中筛选出菌株F4,对该菌株进行耐硒能力测定、鉴定分类、生长还原动力学分析,并通过体外还原试验初步探究亚硒酸钠的还原机理。结果表明:该菌属于高山芽孢杆菌（Bacillus altitudinis）,在浓度为50 mmoL/L亚硒酸钠平板中,菌株菌落存活率为37.08%,对亚硒酸钠的耐受能力可高达250 mmoL/L。在含硒培养基中的还原过程发现,菌株在前期6~14 h先生长然后在对数期中后期14~48 h对亚硒酸钠进行还原,并在稳定时期产生红色沉淀。体外还原定位试验反应体系在33 ℃下孵育24 h后,在培养上清和胞外多糖中观察到反应活性,添加还原型辅酶ⅠNADH下细胞质有略微变化。该芽孢杆菌可作为生产红色纳米硒的菌株,可为硒的生物转化提供参考。     

Distribution of selenium and phenolics in buckwheat plants grown from seeds soaked in Se solution and under different levels of UV-B radiation

Seeds of common buckwheat (Fagopyrum esculentum) were soaked in water, sodium selenate (5, 10 or 20 mg Se^VI/L), or sodium selenite (10 or 20 mg Se^IV/L) solutions. Plants grown from soaked seeds were exposed to reduced UV-B radiation, ambient, or enhanced UV-B. The mass fraction of selenium in leaves was much higher in plants obtained from seeds soaked with selenate (up to 185 ng/g) in comparison to selenite (up to 103 ng/g). In plants obtained from seeds soaked in water, regardless of UV-B levels, the highest concentration of selenium was found in leaves, where the values were between 45 and 66 ng Se/g. In buckwheat leaves 44.5–63.6 mg/100 g d.m. of fagopyrin was found, and in stems 14.3–26.4 mg/100 g d.m.; here no influence of seed soaking solution or UV-B exposure was found. The content of total flavonoids in leaves was 7.8–15.9% and in stems 1.4–4.1%.     

Studies of bioavailability of different food sources of selenium in experiment

The selenium bioavailability in selenium enriched Spirulina (Arthrospira platensis), phycocyanin containing (Se-PC) protein isolate, separated from this micro algae and in sodium selenite was studied and compared in rats. The daily dose of selenium per one animal was 5 microgram in all experimental groups. The average selenium levels in blood serum and liver of animals that received sodium selenite during 14 days were the highest. The average selenium level in blood serum of animals fed with selenium enriched Spirulina platensis after 14 days of receiving was the same with the control group, but the average concentration of selenium in their liver was rather high and close to this parameter of sodium selenite animal group. The animals which were fed with Se-PC showed better results. Their average selenium level in blood serum was higher than in Spirulina group, but lower than in sodium selenite group. The average concentration of selenium in the liver of these animals was the same with sodium selenite animal group. As regards to animals that were fed with selenium enriched Spirulina, Se-PC and sodium selenite for 21 days, the average selenium levels ratio in their blood serum and liver was higher than in control group, but these results were not significantly different among each other. The concentrations of selenium in seminal glands in all groups of animals including control group both after 14 and 21 days feeding were close to each other.     

Selenium speciation in the soil solution and its relevance to plant uptake

The relationship between soil and plant selenium has been investigated using two ⁷⁵SeO₃ spiked silt loam soils from the Hoosfield series with long-term equilibration pH values of 4.5 and 7. Lolium perenne plants were grown in the soils over a 15-week period, and seven harvests were made. Plant samples and soil solutions were assayed for radioactivity, and selenium compounds found in the soil solutions separated by high voltage paper electrophoresis. Throughout the 15-week experimental period selenium speciation was observed to change in soil solution. Samples collected at day 2 from pH 4.5 soil revealed that selenate accounted for 71% and selenite 8% of the soluble selenium compounds present, whereas for the pH 7 soil the relative percentages were 51 and 23% respectively. After 105 days selenate accounted for 22% and selenite 20% at pH 4.5, and 12 and 22% respectively at pH 7. The occurrence of selenoglutathione was noted in the soil solution and its concentration increased significantly with time. Statistically significant correlations (r=0.82) were found between the concentration of selenium in Lolium perenne at harvest and selenate selenium removed from the soil solution.     

Evaluation of the antioxidant capacity of lupin sprouts germinated in the presence of selenium

Antioxidant capacity of 5 DAS (days after seeding) lupin sprouts germinated in the presence of selenate or selenite was determined by cyclic voltammetry (CV), Folin-Ciocalteu reagent (FCR) application and photochemiluminescence (PCL) methods. The hydrophilic (80% methanol) and lipophillic (hexane/methanol) extracts of 5 DAS lupin sprouts germinated in the absence of selenium (control sprouts) showed higher antioxidant capacity than ungerminated seeds. Five DAS sprouts produced in the presence of higher concentration of selenate or selenite (6–8 mg/L) showed increased total antioxidant capacity formed by both hydrophilic and lipohillic antioxidants when evaluated by CV and FCR methods. The hydrophilic extracts from sprouts produced in the presence of low concentration of selenate (2–6 mg/L) showed a significantly higher antioxidant capacity of water-soluble compounds (PCL ACW) while no changes were noted in those germinated in the presence of the highest concentration in relation to the control sprouts. In contrast, the antioxidant capacity of lipid-soluble compounds (PCL ACL) lowered significantly in relation to the control sprouts. The similar changes in antioxidant capacity of sprouts produced in the presence of selenite were found by both PCL assays. Comparison of PCL with CV and FCR reducing capacity assays has shown that these methods yielded considerably different chemical information. Moreover, the changes in total antioxidant and total reducing capacity of sprouts germinated in the presence of either selenate or selenite provided by CV and FCR assays were higher than those obtained by photochemiluminesce measurements. It can be suggested to use 6–8 mg/L of selenite rather than selenate in order to obtain a lupin sprouts with high antioxidant capacity.     

Antagonistic interaction of mercury and selenium in a marine fish is dependent on their chemical species

It is well-known that selenium (Se) shows protective effects against mercury (Hg) bioaccumulation and toxicity, but the underlying effects of Se chemical species, concentration, and administration method are poorly known. In this study, we conducted laboratory studies on a marine fish Terapon jurbua to explain why Hg accumulation is reduced in the presence of Se observed in field studies. When Se and Hg were administrated concurrently in the fish diets, different Se species including selenite, selenate, seleno-dl-cystine (SeCys), and seleno-dl-methionine (SeMet) affected Hg bioaccumulation differently. At high concentration in fish diet (20 μg g(-1) normally), selenate and SeCys significantly reduced the dietary Hg(II) assimilation efficiency (AE) from 38% to 26%. After the fish were pre-exposed to dietary selenite or SeMet (7 μg g(-1) normally) for 22 days with significantly elevated Se body concentrations, the Hg(II) AEs were pronouncedly reduced (from 41% to 15-26%), whereas the dissolved uptake rate constant and elimination rate constant were less affected. In contrast to Hg(II), all the MeHg biokinetic parameters remained relatively constant whether Se was administrated simultaneously with the fish diet or when the fish were pre-exposed to Se with elevated body concentrations. Basic biokinetic measurements thus revealed that Se had direct interaction with Hg(II) during dietary assimilation rather than with MeHg and that different Se species had variable effects on Hg assimilation.     

Mineral substance in human nutrition. Selenium: absorption and bioavailability

In the review there are discussed results of selenium absorption studies in human and animals gastrointestinal tract. According to plenty of experimental data selenate-anion is rapidly and quantitatively absorbed upstream concentration gradient the site of absorption being predominantly ileum. This process is Na+ and energy dependent. On the contrary seleniteanion is absorbed mostly in jejunum, the rate of uptake is less than for selenate in model systems and absorption does not occur toward concentration gradient. Under physiological conditions the most part of selenite is transported as mixed thiols with glutathione and some other tissue thiol compounds. Selenium amino acids are transported by corresponding transfer systems with specificity for analogous sulfur containing amino acids. Whereas bioavailability of transition al metals salts rarely exceeds 10-20%, selenium food sources (both organic and inorganic origin) are typically high absorbable predominantly on 60% or more and in some cases almost on 100%. From this point of view selenium food sources bioavailability evaluation must take into account their metabolic retention and toxicity indicies rather than absorption ratio (such is for zinc, chromium and other metal ions).     

Metabolic pathway for selenium in the body: speciation by HPLC-ICP MS with enriched Se

Selenium (Se) is an ultramicro essential nutrient and both inorganic (selenite and selenate) and organic (selenocysteine and selenomethionine) forms of Se can be used as nutritional sources. Metabolic pathways for Se in the body were studied for selenite and selenate, with the use of enriched 82 Se, by speciation with separation by gel filtration HPLC and detection by element-specific mass spectrometry with ionization with inductively coupled argon plasma (HPLC-ICP MS). The concentrations of 82 Se in organs and body fluids and the distributions of their constituents depending on the dose and time after the intravenous administration of 82 Se-selenite and -selenate to rats were determined. Selenite was taken up by red blood cells within several minutes, reduced to selenide by glutathione, and then transported to the plasma, bound selectively to albumin and transferred to the liver. Contrary to selenite, intact selenate was either taken up directly by the liver or excreted into the urine. The 82 Se of selenite origin and that of selenate origin were detected in the forms of the two Se peak materials in the liver, A and B. The former one was methylated to the latter in vivo and in vitro . The latter one was identical with the major urinary metabolite and it was identified as Se-methyl- N -acetyl-selenohexosamine (selenosugar). The chemical species-specific metabolic pathway for Se was explained by the metabolic regulation through selenide as the assumed common intermediate for the inorganic and organic Se sources and as the checkpoint metabolite between utilization for the selenoprotein synthesis and methylation for the excretion of Se.