Morphological and physiological changes exhibited by a Cd-resistant Dictyosphaerium chlorelloides strain and its cadmium removal capacity

Changes induced on freshwater microalga Dictyosphaerium chlorelloides (Dc^wt) acclimated in the laboratory until their survival in culture media enriched with cadmium 100 µM have been studied. Cadmium removal by living cells of this Cd-resistant (Dc^CdR100) strain was tested in cultures exposed to 100 µM Cd during 30 days. Cell dimensions were measured under light microscopy, and cell growth was studied. Photosynthetic yield (Φ_PSII) was analyzed and the photosynthetic oxygen development and respiration response was obtained. Results show that Dc^CdR100 strain exhibited significant cell morphology changes in comparison to Dc^wt cells, which affected both surface area and cell biovolume. Malthusian fitness analysis showed that Dc^CdR100 strain living in Cd-enriched culture had developed a lower capacity of nearly 50% growth, and its photosynthetic oxygen development and respiration response were significantly reduced in both light and dark photosynthetic phases. Dc^CdR100 strain showed a very high capacity to remove cadmium from the aquatic environment (over 90%), although most of the removed heavy metal (≈70%) is adhered to the cell wall. These specific characteristics of Dc^CdR100 cells suggest the possibility of using this strain in conjunction with Dc^wt strain as bioelements into a dual-head biosensor, and in bioremediation processes on freshwater polluted with Cd.     

Tissue cadmium (Cd) concentrations of people living in a Cd polluted area, Japan

There are more than 50 cadmium (Cd) polluted areas in Japan. The severest general environmental Cd polluted area in Japan was the Jinzu River basin in Toyama Prefecture where Itai-itai disease had been endemic. The present study aimed to compare organ Cd concentrations of inhabitants who had been exposed to different levels of environmental Cd and to clarify the health effects of the environmental Cd exposure in Japan. Since 1960 we have measured tissue Cd concentrations of inhabitants with a history of living in a different Cd polluted areas. Study population living in Cd polluted areas were 36 (1 male, 35 females) patients with Itai-itai disease, 20 (7 males, 13 females) subjects suspected of having Itai-itai disease, 8 (2 males, 6 females) inhabitants in Cd polluted areas other than the Jinzu River basin. Subjects who had lived in Cd non-polluted area were 72 inhabitants. Cd concentrations in liver, pancreas and thyroid of those living in Cd polluted areas were as high as those of patients with Itai-itai disease, and their Cd concentrations in renal cortex were as low as those of patients with Itai-itai disease. The present study demonstrated that tissue Cd concentrations of some inhabitants in Cd polluted areas other than Jinzu River basin are equal to those of the patients with Itai-itai disease and that patients with Itai-itai disease were present even in these areas.     

Cadmium removal by a new strain of Pseudomonas aeruginosa in aerobic culture.

A fluorescent pseudomonad (strain CW-96-1) isolated from a deep-sea vent sample grew at 30 degrees C under aerobic conditions in an artificial seawater medium and tolerated cadmium concentrations up to 5 mM. After 140 h, strain CW-96-1 removed > 99% of the cadmium from solution. Energy dispersive microanalysis revealed that the cadmium was removed by precipitation on the cell wall; sulfide production was confirmed by growth on Kligler's agar. Based on 16S ribosomal DNA sequencing and fatty acid analysis, the microorganism is closely related to Pseudomonas aeruginosa.     

Conjugation in Polytomella agilis Araga˜o (Volvocida,Polyblepharidae)*

SYNOPSIS. The occurrence of conjugation in the flagellate, Polytomella agilis Aragão, as first reported in 1910, is confirmed. The gametes made contact with each other in a variety of positions. Our observations differ in minor ways from those reported for P. agilis and are in considerable disagreement with others published on sexual reproduction in Polytomella caeca.     

两株多环芳烃降解菌协同对菲-镉污染的去除特性北大核心

【目的】从污染土壤中分离筛选一株多环芳烃降解菌,并探究其与Pseudomonas aeruginosa B6-2构建的混菌体系对菲-镉复合污染的修复效能,以及微生物代谢特性对不同镉浓度赋存的响应特性,以期为复合污染的生物修复提供优良菌株资源及应用技术参考。【方法】采用富集驯化、筛选纯化方法得到一株多环芳烃降解菌,通过生理生化特征和16S rRNA基因序列分析进行鉴定。利用高效液相色谱法和电感耦合等离子体质谱法评估不同镉浓度赋存下各反应体系对菲和镉的去除效能;通过菌体细胞形态的扫描电镜观测及菌株代谢活性检测,探讨镉胁迫对菲生物降解过程的影响机制。【结果】筛选得到一株具有重金属耐受性和多环芳烃高效降解菌SZ-3,经鉴定为节杆菌属;降解菌协同体系(M)具有良好的菲降解效能和抗镉胁迫优势。镉胁迫浓度为0.5、10 mg/L时,M对菲和镉的去除率分别高于85%、80%;镉胁迫浓度为25、50 mg/L时,2种污染物的去除率均大于65%。扫描电镜分析表明,镉胁迫导致菌体表面粗糙且出现不同程度变形,菌体间黏附性和聚集性提高。反应周期内,邻苯二酚1,2-双加氧酶活性与电子传递体系活性随镉浓度增加而降低,两者变化与菲降解速率变化一致。【结论】Arthrobacter sp.SZ-3是一株PAHs高效降解菌,能与Pseudomonas aeruginosa B6-2协同高效修复菲-镉复合污染,随着初始镉胁迫浓度增加,混菌协同对目标污染物去除的优势显著。     

Cadmium removal in a biosorption column

New biosorbent material derived from a ubiquitous brown marine alga Ascophyllum nodosum has been examined in packed-bed flow-through sorption columns. It effectively removed 10 mg/L of cadmium down to 1.5 ppb levels in the effluent, representing 99.985% removal. The experimental methodology used was based on the early Bohart and Adams sorption model, resulting in quantitative determination of the characteristic process parameters which can be used for performance comparison and process design. An average metal loading of the biosorbent (N(0)) determined was 30 mg Cd/g, corresponding closely to that observed for the batch equilibrium metal concentration of 10 mg Cd/L. The critical bed depth (D(min)) for the potable water effluent quality standard (0.005 mgg Cd/L) varied with the column feed flow rate (2.4 to 9.6 L/h . cm(2)) from 20 to 50 cm. The sorption column mass transfer and dispersion coefficients were determined, which are also required for solving the sorption model equations. (c) 1994 John Wiley & Sons, Inc.     

好氧反硝化生物脱氮技术的研究进展

好氧反硝化生物脱氮技术自提出以来,凭借能实现同步硝化反硝化、节省基建投资及运行费用等诸多优点,受到国内外环境领域学者的广泛关注。本文首先总结了近年来好氧反硝化菌种的筛选分离情况,以及环境因子对好氧反硝化菌脱氮效能的影响,包括溶解氧(dissolved oxygen,DO)、碳氮比(C/N)、温度等。然后深入探讨了好氧反硝化生物脱氮技术的原理,好氧反硝化过程中的关键功能基因及酶,同时介绍了分子生物技术在好氧反硝化研究过程中的应用,以及好氧反硝化生物脱氮技术在实际应用方面的研究现状。最后,基于目前的研究瓶颈问题,对未来好氧反硝化生物脱氮技术的研究方向提出了科学展望。     

Simultaneous COD, nitrogen, and phosphate removal by aerobic granular sludge

Aerobic granular sludge technology offers a possibility to design compact wastewater treatment plants based on simultaneous chemical oxygen demand (COD), nitrogen and phosphate removal in one sequencing batch reactor. In earlier studies, it was shown that aerobic granules, cultivated with an aerobic pulse-feeding pattern, were not stable at low dissolved oxygen concentrations. Selection for slow-growing organisms such as phosphate-accumulating organisms (PAO) was shown to be a measure for improved granule stability, particularly at low oxygen concentrations. Moreover, this allows long feeding periods needed for economically feasible full-scale applications. Simultaneous nutrient removal was possible, because of heterotrophic growth inside the granules (denitrifying PAO). At low oxygen saturation (20%) high removal efficiencies were obtained; 100% COD removal, 94% phosphate (P-) removal and 94% total nitrogen (N-) removal (with 100% ammonium removal). Experimental results strongly suggest that P-removal occurs partly by (biologically induced) precipitation. Monitoring the laboratory scale reactors for a long period showed that N-removal efficiency highly depends on the diameter of the granules.     

Paternity success and the direction of sexual selection in a field population of a semelparous marsupial,Antechinus agilis

Antechinus agilis is a small sexually size dimorphic marsupial with a brief annual mating period of 2-3 weeks. All males die after this period, and females give birth to up to 10 young. Mating is thought to be promiscuous, however, there is no field data to confirm this. Using microsatellites, we investigated paternity patterns over two seasons in a wild population. Male weight was significantly positively related to the number of females fertilized and with the number of offspring sired, in both years. Furthermore, selection gradients indicated selection for larger males. Both results suggest that size dimorphism in A. agilis can be explained by sexual selection for larger males. The proportion of offspring sired within litters, did not relate to male size. Therefore, larger males are more successful through higher mating access, not through their sperm outcompeting that of smaller males. As expected from their known ranging behaviour, the number of offspring within litters left unassigned to a father did not depend on the grid location of the mother. Female size did not differ between successful reproducing and unsuccessful females. However, females that weaned offspring had larger heads than females that did not wean offspring. Males did not 'prefer' mating with larger females, nor did assortative mating occur. From our results, the mating system of A. agilis is clearly promiscuous. Selection for larger males occurred in both years, even though in one year the operational sex ratio was highly female biased, suggesting that the potential reproductive rate is a better predictor of the direction of sexual selection in A. agilis.     

Implication of distinct proteins in cadmium uptake and transport by intestinal cells HT-29

The mechanisms of intestinal absorption have not been clearly elucidated for cadmium, a toxic metal. In this work, we show the implication of distinct proteins in cadmium transport, and the transport step where these proteins are involved. We first validated the HT-29 model by evaluating nontoxic doses of cadmium (ranging from 1 to 20 μmol/L), and by quantifying metal uptake and transepithelial transport. The time-course of 1 μmol/L cadmium uptake at pH 7.5 showed three steps: a rapid one during the first 4 min, probably due to cadmium binding to the membrane; a slower one, characterized by K _m of 1.65±0.54 μmol/L and V _max of 3.9±0.3 pmol/min per mg protein; and a third, corresponding to slow accumulation that was not equilibrated even after 48 h of cadmium exposure. Intracellular metallothionein content following 1 or 5 μmol/L cadmium exposure showed a significant increase after 6 h of exposure, and was not equilibrated even after 72 h, allowing cadmium accumulation. After 24 h of exposure, metallothionein content was 5-fold, 14-fold, 26-fold, and 50-fold, respectively, for cells grown in the presence of 1, 5, 10, and 20 μmol/L cadmium, compared to control cells. The second step of uptake, characterized by carrier-mediated transport, was markedly increased at pH 5.5, compared to pH 7.5, and strongly inhibited by the metabolic inhibitor dinitrophenol. Moreover Nramp2 transporter cDNA was present in HT-29 cells. These data suggest the involvement of a proton-coupled transporter, which may be the divalent cation transporter Nramp2 (natural resistance-associated macrophage protein 2). Cadmium uptake was also inhibited by copper, zinc, and para-chloromercuribenzenesulfonate (pCMBS), but not by verapamil or ouabain. Taken together, our results indicate that cadmium could enter HT-29 cell by Nramp2 proton-coupled active transport and by diffusion, and accumulates in the cell as long as it binds to metallothionein. Cadmium toxicity could depend partly on the activity of Nramp2, and partly on metallothionein content. This revised version was published online in July 2006 with corrections to the Cover Date.     

镁、锰、活性炭和石灰及其交互作用对小麦镉吸收的影响

采用盆栽试验,研究了在镉污染土壤上施用石灰、硫酸镁、硫酸锰和活性炭不同用量以及交互作用对小麦生长和吸收重金属镉的影响.研究结果表明,在试验条件下施用适量的硫酸镁、硫酸锰或与石灰配合能明显提高小麦籽粒产量,单施石灰或与活性炭配合施用降低了小麦籽粒产量;与对照(CK)相比,所有处理秸秆产量均下降.施用硫酸镁能显著降低小麦籽粒和秸秆中Cd浓度,且随用量的增加两增大.低量硫酸锰能有效降低小麦籽粒和秸秆中Cd浓度,高量反而增加小麦对Cd的吸收.石灰、活性炭单独施用或配合施用都能明显减少小麦对Cd的吸收,但籽/杆中Cd比却随石灰用量的增加呈明显的上升趋势.叶面喷施硫酸镁对降低小麦吸收镉的效果与土施相当,但叶面喷施硫酸锰却比土施硫酸锰显著降低了小麦籽粒中的镉浓度与吸收量.硫酸镁与硫酸锰,或石灰、硫酸镁和硫酸锰3种物质配合施用,对小麦籽粒镉浓度和吸收量的降低表现出明显的正交互作用,对抑制小麦体内镉从秸秆向籽粒的转移具有显著效果.     

Increased metal tolerance and bioaccumulation of zinc and cadmium in Chlamydomonas reinhardtii expressing a AtHMA4 C-terminal domain protein

The use of microalgal biomass for metal pollutant bioremediation might be improved by genetic engineering to modify the selectivity or capacity of metal biosorption. A plant cadmium (Cd) and zinc (Zn) transporter (AtHMA4) was used as a transgene to increase the ability of Chlamydomonas reinhardtii to tolerate 0.2 mM Cd and 0.3 mM Zn exposure. The transgenic cells showed increased accumulation and internalization of both metals compared to wild-type. AtHMA4 was expressed either as the full-length (FL) protein or just the C-terminal (CT) tail, which is known to have metal-binding sites. Similar Cd and Zn tolerance and accumulation was observed with expression of either the FL protein or CT domain, suggesting that enhanced metal tolerance was mainly due to increased metal binding rather than metal transport. The effectiveness of the transgenic cells was further examined by immobilization in calcium alginate to generate microalgal beads that could be added to a metal contaminated solution. Immobilization maintained metal tolerance, while AtHMA4-expressing cells in alginate showed a concentration-dependent increase in metal biosorption that was significantly greater than alginate beads composed of wild-type cells. This demonstrates that expressing AtHMA4 FL or CT has great potential as a strategy for bioremediation using microalgal biomass.     

Metal uptake by different species of phytoplankton in culture

Cadmium and copper uptake by cultures of Prasinocladus marinus, Amphidinium carterae, Chaetoceros curvisetum and Chaetoceros protuberans was considered as a function of exposure time (from 0.5 to 72 h) and of metal concentrations added to the culture medium (from 5 to 20 µg l^–1). For cadmium uptake, A. carterae exhibited a particular behaviour: the accumulation was not dependent on exposure time or on metal concentration. An adsorption process appeared to govern both Cadmium uptake by phytoplankton species (except A. carterae) and that of copper. In addition to adsorption, another active process might be involved in the Cadmium uptake of P. marinus, C. curvisetum and C. protuberans.     

Improvement of Cd2+ uptake ability of SmtA protein by Lys/Cys mutation; experimental and theoretical studies

The improved Cd²⁺ surface affinity characteristics of a mutated cyanobacterial metallothionein SmtA (K45C) were investigated via experimental and theoretical methods. Molecular dynamics simulations were carried out using a model of Cd²⁺ and other ions enclosed in a fully hydrated simulation box with the wild-type or mutated SmtA protein. The theoretical results suggested that mutated SmtA was more powerful in absorption of Cd²⁺ than the wild-type protein. Then, the mutated smtA gene (from Synechococcus PCC 7942) was synthesized by simplified gene synthesis method and expressed on isopropyl-beta-d-thiogalactopyranoside induction. The protein expression was investigated by SDS-PAGE and verified by Western blotting. Finally, cadmium uptake ratio of mutant protein toward wild type was analyzed by atomic absorption. This study is the first example of cytoplasmic expression of a mutant protein. Experimental results also verified that the mutation intensifies uptake of Cd²⁺ ions.     

Effect of linear alkylbenzene sulfonate on Cu2+ removal by Spirulina platensis strain (FACHB‐834)

The removal efficiency of Cu²⁺ by Spirulina platensis (strain FACHB‐834), in viable and heat‐inactivated forms, was investigated in the presence and absence of linear alkylbenzene sulfonate (LAS). When the initial Cu²⁺ concentration was in the range of 0.5–1.5 mg · L^?1, a slight increase in growth rate of FACHB‐834 was observed. In contrast, when Cu²⁺ or LAS concentrations were at or higher than 2.0 or 6.0 mg · L^?1, respectively, the growth of FACHB‐834 was inhibited and displayed yellowing and fragmentation of filaments. The presence of LAS improved Cu²⁺ removal by ~20%, and accelerated attainment of Cu²⁺ retention equilibrium. For the 2‐ mg · L^?1 Cu²⁺ treatments, retention equilibrium occurred within 2 d and showed maximum Cu²⁺ removal of 1.83 mg · L^?1. In the presence of LAS, the ratio of extracellular bound Cu²⁺ to intracellular Cu²⁺ taken up by the cells was lower (1.05–2.26) than corresponding ratios (2.46–7.85) in the absence of LAS. The percentages of extracellular bound Cu²⁺ to total Cu²⁺ removal (both bound and taken up by cells) in the presence of LAS ranged from 51.2% to 69.3%, which was lower than their corresponding percentages (71.1%–88.7%) in the absence of LAS. LAS promoted biologically active transport of the extracellular bound form of Cu²⁺ into the cell. In contrast, the addition of LAS did not increase the maximum removal efficiency of Cu²⁺ (61.4% ± 5.6%) by heat‐inactivated cells compared to that of living cells (59.6% ± 6.0%). These results provide a theoretical foundation for designing bioremediation strategies using FACHB‐834 for use in surface waters contaminated by both heavy metals and LAS.     

Effects of cadmium on the uptake,distribution and assimilation of nitrate in Pisum sativum

The net uptake, distribution and assimilation of NO ₃ ^– were studied in pea plants subjected to either long-term continuous Cd treatment for 10 d (10 or 50 M Cd) or short-term treatment (72 h) with 50 M Cd. In the latter treatment, the effects of transferring the plants to a Cd-free nutrient solution for a 'recovery period' of 96 h were also studied. All these treatments were compared with 'controls', plants which received no Cd. In both experiments, the reduction in fresh weight was associated with a decrease in the content (%) of shoot and root water and in transpiration rate as Cd concentration increased. The concentration of ₃ ^– in the shoots and sap decreased dramatically and net ₃ ^– uptake was severely inhibited, effects associated with a loss of shoot nitrate reductase (NR) activity. In the short-term Cd treatment, net ₃ ^– uptake was almost completely inhibited after 24 h, but recovered after the transfer of plants to a Cd-free nutrient solution. Similarly, a dramatic decrease in the shoot NR activity was observed. The uptake, distribution and tissue partitioning of K was also studied, which is considered to be the major counterion of ₃ ^– . Potassium uptake was similarly affected by Cd, as inferred from the ratio ₃ ^– /K uptake, which was ca. 10. The ratio K/ ₃ ^– tissue content increased in the shoot concomitantly to Cd in both long-term and short-term metal supply. These parameters showed a tendency of K similar to that observed for ₃ ^– , although its relative tissue distribution was not affected by Cd.     

Improved phosphate removal by selective sludge discharge in aerobic granular sludge reactors

Two lab-scale aerobic granular sludge sequencing batch reactors were operated at 20 and 30°C and compared for phosphorus (P) removal efficiency and microbial community composition. P-removal efficiency was higher at 20°C (>90%) than at 30°C (60%) when the sludge retention time (SRT) was controlled at 30 days by removing excess sludge equally throughout the sludge bed. Samples analyzed by fluorescent in situ hybridization (FISH) indicated a segregation of biomass over the sludge bed: in the upper part, Candidatus Competibacter phosphatis (glycogen-accumulating organisms--GAOs) were dominant while in the bottom, Candidatus Accumulibacter phosphatis (polyphosphate-accumulating organisms--PAOs) dominated. In order to favour PAOs over GAOs and hence improve P-removal at 30°C, the SRT was controlled by discharging biomass mainly from the top of the sludge bed (80% of the excess sludge), while bottom granules were removed in minor proportions (20% of the excess sludge). With the selective sludge removal proposed, 100% P-removal efficiency was obtained in the reactor operated at 30°C. In the meantime, the biomass in the 30°C reactor changed in color from brownish-black to white. Big white granules appeared in this system and were completely dominated by PAOs (more than 90% of the microbial population), showing relatively high ash content compared to other granules. In the reactor operated at 20°C, P-removal efficiency remained stable above 90% regardless of the sludge removal procedure for SRT control. The results obtained in this study stress the importance of sludge discharge mainly from the top as well as in minor proportions from the bottom of the sludge bed to control the SRT in order to prevent significant growth of GAOs and remove enough accumulated P from the system, particularly at high temperatures (e.g., 30°C).     

Specific oxygen, ammonia, and nitrate uptake rates of a biological nutrient removal process treating elevated salinity wastewater

Anaerobic/anoxic/aerobic systems inoculated without and with NaCl acclimated cultures, i.e., Models A and B, respectively, were fed with a synthetic wastewater at various salinity levels. After achieving a steady state, the systems were shocked with 70 g/l NaCl for four consecutive days before returning to pre-shock conditions. At the steady-state, the specific oxygen uptake rates (SOURs) increased with an increase of sodium chloride concentration (from 5.40 to 9.72 mg O₂/g mixed liquor suspended solids (MLSS)-h at 0–30 g/l NaCl for Model A and from 6.84 to 17.64 mg O₂/g MLSS-h at 5–30 g/l NaCl for Model B). In contrast, the specific ammonia uptake rate (SAUR) and specific nitrate uptake rate (SNUR) decreased with increasing chloride concentration (from 4.76 to 2.14 mg NH₃–N/g MLSS-h and 2.50 to 1.22 NO3–N/g MLSS-h, for Model A, and from 3.84 to 2.71 mg NH₃–N/g MLSS-hr and 2.54 to 1.82 mg NO₃–N/g MLSS-hr, for Model B). During the shocked period, the SOUR in most scenarios increased whereas the SAUR and SNUR tended to decrease. The impact of the chloride shock on nitrifiers was more obvious than on denitrifiers; however, after a certain recovery period, the activities of both nitrifiers and denitrifiers in terms of SAUR and SNUR were approximately the same as those prior to shock.     

Biosorption of cadmium,cobalt, nickel,and strontium by aBacillus simplex strain isolated from the vadose zone

A subsurface Gram-positive, endospore-forming, filamentous bacterium, designated ZAN-044, was isolated from a depth of 96.2 m in the vadose zone of the Hanford Site in Washington State. A phylogenetic analysis of the 16S rRNA gene sequence of strain ZAN-044 revealed it to be 99.5% similar toBacillus simplex strain DSM 1321, indicating that they may be members of the same species.B. simplex ZAN-044 was studied along withBacillus subtilis 168, andEscherichia coli K-12 (AB264), two well-characterized metal-sorbing bacteria, for the binding of Cd²⁺, Co²⁺, Ni²⁺, and Sr²⁺. There was rapid (less than 1 h) uptake of 1 M metal by the three bacteria in the order Cd>NiCo>Sr. Binding followed a saturation isotherm at cation concentrations from 0.1 M to 1 mM. Cation binding was pH-dependent, with less binding at low pH.B. simplex ZAN-044 bound more metal thanB. subtilis orE. coli, demonstrating that subsurface microorganisms can remove significant quantities of metals from solution and may be able to influence radionuclide and metal transport in the subsurface.