光合细菌的分离、鉴定和固定化及其在净化鱼池水质中的应用研究

本文报告了光合细菌的分离、鉴定及固定在载体ＰＶＡ上,用于鲤鱼育苗池中净化水质的研究结果。并验分别用固定化光合细菌、游离光合细菌和对照三组,在三个鱼池中分别放养７００尾鱼苗,每５日测鱼池水中氨态氮一次,共测６次,其结果表明,应用固定化光合细菌的鱼池、氨的去除率高达９０％以上,有明显净化鱼池水质的作用,从而有利于鱼苗的生长。     

复合固定化光合细菌及其处理养鱼水的效果

利用海藻酸钠和沸石,将含有球形红假单胞菌、荚膜红假单胞菌、沼泽红假单胞菌、万尼氏红微菌等菌体的复合光合细菌固定化,研究其对养鱼水的氮磷去除效果.比较了两种不同包埋材料固定化光合细菌处理养鱼废水的效果,对固定化光合细菌去除废水中氮磷的工艺条件进行了优化、并通过生物反应器连续处理养鱼水分析了处理后水质的效果.通过2 种固定化工艺的比较,确定了2 %沸石+2 %海藻酸钠(CA )的凝胶剂组合作为固定材料,其颗粒内生物活性最高.复合固定化光合细菌处理养鱼水的最佳条件为:厌氧光照条件下,颗粒粒径3 mm ,包埋比1 :5 ,颗粒投加量5 mg·L-1 ,4d 后养鱼水中NH4+-N 、PO43-和CODMn 的去除率分别为74.4%、84.26%、78.92%.此外,通过连续试验可以看出,固定化光合细菌具有明显的去除氨氮、磷酸盐的作用,其在净化养鱼水质方面具有非常明显的优越性.     

高效净水异养硝化细菌的筛选鉴定及固定化应用研究

本研究从海参养殖水体、泥土中筛选出4株具有硝化能力的异养硝化细菌。分别将其游离菌体细胞投入海参养殖水体,测定亚硝态氮、氨氮去除率,筛选出HS．NOB2为高效净化菌株,对HS-NOB2进行16SrDNA扩增及序列测定,初步鉴定为节杆菌（Arthrobactersp．）。利用海藻酸钠包埋法对高效净化菌体细胞进行固定化,将该固定化菌投入养殖水体及人工合成污水,研究其对水体中亚硝态氮、氨氮的处理效果,并与游离菌体细胞进行比较。结果表明,固定化后亚硝态氮去除率达到49．85％,氨氮去除率达到56．58％,均明显高于游离菌体细胞。上述研究为探寻水体净化提供了新思路,为水质改良剂的实际生产提供可选菌株。     

固定化微生物菌剂与蕹菜对中华鳖养殖污水的净化效果研究

为解决中华鳖(Trionyx sinensis)养殖所造成的水环境污染问题,研究利用海藻酸钠、沸石对复合光合菌剂和枯草芽孢杆菌菌剂分别进行固定化,并研究了不同固定化微生物菌剂与蕹菜对中华鳖养殖污水的净化效果。结果表明:净化处理15d后,利用固定化微生物菌剂与蕹菜共同处理组对中华鳖养殖污水的净化处理效果最好,养殖污水的CODMn、NH 4+-N、NO3-N、NO 2-N、PO34的去除率分别为88.7%、87.3%、90.8%、98.3%、74.9%。蕹菜处理组对于NO3-N、NO 2-N、NH 4+-N的去除率显著高于固定化复合光合菌剂处理组和固定化枯草芽孢杆菌处理组。固定化复合光合菌剂处理组对于NO3-N、NO 2-N、NH 4+-N的去除率显著高于固定化枯草芽孢杆菌菌剂处理组,但固定化枯草芽孢杆菌处理组对于污水的CODMn的去除率最高,可达90.6%。     

固定化技术在促进光合细菌产氢中的应用

氢气是一种新型的清洁高效能源,制氢技术的创新是目前研究的热点。将新型的技术及材料应用到生物制氢工艺中,从而促进生物制氢技术的产氢效率和工程应用是研究的重点之一。该文阐述了光合细菌在固定化生长条件下发酵产氢的最新研究进展,从固定化技术的原理、固定化方法的应用进展及影响因素几个方面进行了综述,详细阐述了包括包埋、悬浮载体附着生长及固定生物膜法等几种固定化方法对光发酵产氢的作用,介绍了国内外用于固定化的新型材料,并对今后的研究重点及方向进行了展望。     

净化养殖水体紫色非硫光合细菌的筛选与鉴定

本研究从杭州养鱼塘水样及底泥样品中富集分离得到三株紫色非硫光合细菌,分别命名为HZ-3,HZ-4,HZ-5。 通过比较这三株菌对鱼、虾养殖水体的净化效果,筛选出菌株HZ-5 净化能力较强 ,经过5天的处理,该菌株使养鱼塘水样的COD 降低24.87%;使养虾池塘水样的COD 降低 36.99%;使养鱼塘水样的亚硝态氮的降解率达到97.79%。对菌株HZ-5进行了形态学观察、生理生化鉴定、活细胞吸收光谱以及16S rDNA序列分析。16S rDNA序列分析结果表明该菌株与沼泽红假单胞菌的16S rDNA序列有高达99％的同源性,结合形态特征和生理生化特性以及活细胞吸收光谱特征等,将其鉴定为沼泽红假单胞菌（Rhodopseudomonas palustris）。     

光合细菌在水污染治理中的研究进展

光合细菌以其无毒、繁殖快、适应能力强、易人工培养等优点而在环境治理中受到重视。国内外对光合细菌的研究主要集中在水产养殖业(如净化水质,作饵料添加剂等)和生活及工业重污染水处理中的作用,关于光合细菌在景观微污染水体治理方面的作用研究较少。课题组研究发现光合细菌中的沼泽红假单胞菌对西南大学景观水中氨氮的去除率高达95%,暗示光合细菌能有效治理景观水污染。综述了光合细菌的分类、脱氮除磷原理以及目前光合细菌在治理有机废水、重金属废水和养殖污水方面的应用,并展望了光合细菌在处理景观微污染水体方面的应用前景,以期为进一步研究光合细菌在景观水治理中的作用提供参考。     

聚氨酯泡沫半固定化培养盘基网柄菌

研究了聚氨酯泡沫应用于固定化盘基网柄菌的可行性,发现以简单处理过的聚氨酯泡沫为载体,能够高效实现盘基网柄菌的固定化培养。考察了载体粒径大小、载体量和摇床转速等对固定化培养的影响,在优化的培养条件和固定化条件下,盘基网柄菌的最大细胞密度是悬浮培养的2~4倍。     

光合细菌的分离、保存及其同化磷能力试验研究

为了研究光合细菌的保藏方法及其同化磷能力,采用液体种室内自然放置、液体种蜡封和穿刺物蜡封等方法对光合细菌进行保藏方法研究,结果表明,液体种蜡封和穿刺物蜡封可作为该菌种的长期保存方法;对两株菌的磷同化能力进行了比较,降解率分别为22%和15%。本研究为获得光合细菌的长期保存方法及其在污水净化中的应用提供参考。     

光合细菌的分离、保存及其同化磷能力研究

为了研究光合细菌的保藏方法及其同化磷能力,采用液体种室内自然放置、液体种蜡封和穿刺物蜡封等方法对光合细菌进行保藏方法研究,结果表明,液体种蜡封和穿刺物蜡封可作为该菌种的长期保存方法;对两株菌的磷同化能力进行了比较,降解率分别为22%和15%。本研究为获得光合细菌的长期保存方法及其在污水净化中的应用提供参考。     

Comparative studies of physiological and environmental effects on the production of cyclosporin A in suspended and immobilized cells of Tolypocladium inflatum

Process improvement of the production of cyclosporin A (Cy A), a powerful immunosuppressive fungal metabolite, has been undertaken by analyzing suspended and immobilized cell cultures in parallel. Conidiospores of the producer microorganism, Tolypocladium inflatum, were entrapped into porous celite particles. Easier germination of the entrapped spores and more active growth of the immobilized cells were manifested when compared with free cell cultures initiated with spores or with mycelial inocula. Significant differences in precursor flow between the immobilized and free cell systems were evident when the effects of L-valine (a constituent amino acid of the Cy A molecule) on Cy A biosynthesis were compared in the two systems. For the freely suspended cells, L-valine supplemented early in the fermentation served as a possible precursor or stimulator of Cy A biosynthesis. A significant increase in specific production and Cy A yield on carbon source was observed in this system relative to suspended cultures supplemented with L-valine during or after exponential growth. In contrast to the free cell cultures, the addition of L-valine during the initial stage of immobilized cell growth had a negative effect on Cy A production but resulted in somewhat increased cell growth. This suggests an incompatibility between primary and secondary metabolic networks involved in Cy A biosynthesis in the immobilized state upon external addition of the amino acid.     

Effects of high cell density on growth-associated monoclonal antibody production by hybridoma T0405 cells immobilized in macroporous cellulose carriers

Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production of hybridoma T0405 cells immobilized in macroporous cellulose carriers were investigated in continuous and batch cultures. The results showing, that the specific MAb production rate increased with increasing specific growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. Moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomena, MAb mRNA expression and cell cycle distribution were investigated in batch cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromode-oxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb mRNA expression reached the peak during the exponential growth phase, suggest a positively growth-associated MAb production. And the immobilized cells continued the MAb mRNA expression until dead phase, which was longer than that in suspended cells. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated MAb productivity of T0405 cells.     

Media and environmental effects on phenolics production from tobacco cell cultures

The effects of various factors in culture medium on the phenolics production from cultured tobacco cells (free and immobilized) were studied. It was found that removing the growth hormone from the medium increased the productivities of phenolics for both free and immobilized cultures. Low initial sucrose concentration in the medium restricted growth and resulted in high cellular productivities of the phenolics for freely suspended cells, but not for the immobilized cells. Addition of 1.4% DMSO to standard culture medium greatly increased phenolics productivities without affecting cell viability in both free and immobilized cell cultures. Continuous operation of a packed-column reactor of the immobilized cells was achieved for 500 hours. Aeration was accomplished by diffusing pure oxygen through silicone tubing placed inside the reactor. It was found that prolonged cell viability was contingent upon initially operating the reactor with total recycle for several days, and then introducing fresh feed while maintaining a high recycle rate. Immobilized cells packed in a continuous column reactor achieved productivities more than twice that achieved in any batch run.     

Thermostability of immobilized plant microsomes

Microsomes from grapewine (V. vinifera L. cv. Camay Fréaux) cell homogenates were immobilized in calcium alginate beads. Cinnamic acid-4-hydroxylase activities for both freely suspended and immobilized microsomes were compared, resulting in a decrease of 30–40% of the initial activity as a consequence of immobilization. This decrease was probably due to both protein denaturation and microsomal mass losses during the gelation process. Nevertheless, immobilized microsomes showed an enhanced thermostability. Only 35% of the initial activity was lost when immobilized microsomes were incubated at 45°C for 10 minutes, whereas a 68% activity loss was observed when dealing with free microsomes.     

光合细菌对水产养殖水质和水生生物的影响

研究光合细菌对水产养殖水体的水质,如化学需氧量(COD)、氨氮(NH₄⁺-N)、溶解氧(DO)、pH等指标,及对水生生物,如藻类植物和细菌的种群组成的影响。结果表明:鲤鱼养殖水中投放一定量的光合细菌,能明显去除水中有机物和NH₄⁺-N,增加DO的含量,稳定pH值。光合细菌对水中藻类也有明显影响,0.50%、1%剂量组中藻类明显增加,其中硅藻在所有藻类中所占比例可达26%以上,绿藻所占比例达60%左右,而以蓝藻为主的杂藻则下降到10%左右。光合细菌对有害菌有一定的抑制作用,使得弧菌属、气单胞菌属等致病菌所占比例显著降低。     

不同接种浓度绿球藻对水产养殖废水净化的影响

通过设置绿球藻(Chlorococcum sphacosum GD)的起始接种浓度(25—400 mg/L),研究其对水产养殖废水的处理效果及藻细胞的生长特性。研究结果表明,起始接种浓度为100 mg/L的绿球藻藻液,其生长特性最佳,比生长速率最大,倍增时间最短。随着起始接种浓度的增加,生长速率逐渐降低,倍增时间逐渐增加。在起始接种浓度为100 mg/L的条件下,在5d的培养周期内,绿球藻能够去除水产养殖废水中96.92%的COD、98.08%的氨氮、98.67%的亚硝氮、91.42%的硝氮及98.36%的总磷。低起始接种浓度(尤其是100 mg/L)有利于绿球藻的生长和污染物降解。研究初步探明了微藻起始接种浓度对水产养殖废水处理效果的影响。通过控制微藻接种浓度有望在提高污染物去除率的同时缩短培养周期并提高容积负荷,为今后微藻用于大规模水产养殖废水的处理提供了一定的理论支持。     

Production of rifamycin B and SV by free and immobilized cells of Amycolatopsis mediterranei

The effect of growth conditions (incubation time, inoculum size, initial pH value) and some nutrient concentrations on the growth and rifamycin B and SV production by free and immobilized cells of Amycolatopsis mediterranei CBS 42 575 was studied. In alginate beads, the immobilized cells behaved like the free cells, but a pronounced difference was observed in antibiotic production and cell growth. The rifamycin production by the immobilized cells was higher than that obtained by the free cells. The immobilized cells were also reused repeatedly for six batch cultivations with a fresh medium charged into flasks at the beginning of each batch. It was found that the immobilized cells were stable, and the rifamycin yield was almost constant during the first three batches and then decreased.     

Induction of purple sulfur bacterial growth in dairy wastewater lagoons by circulation

Aims:  To determine whether circulation of dairy wastewater induces the growth of phototrophic purple sulfur bacteria (PSB).
Methods and Results:  Two dairy wastewater lagoons that were similar in size, geographic location, number and type of cattle loading the lagoons were chosen. The only obvious visual difference between them was that one was stagnant and the water was brown in colour (Farm 1), and the other was circulated and the water was red in colour because of the presence of PSB that contained carotenoid pigments (Farm 2). Both wastewaters were sampled monthly for 3 months and assayed for PSB and extractable carotenoid pigments (ECP). After this point, circulators were placed in the wastewater lagoon on Farm 1, and samples were taken monthly for 9 months and assayed for PSB and ECP. Before the installation of circulators, no PSB-like 16S rRNA sequences or ECP were observed in the wastewater from Farm 1; however, both were observed in the wastewater from Farm 2. After the installation of circulators, statistically greater levels of PSB and extractable carotenoid pigments were observed in the wastewater from Farm 1.
Conclusions:  Circulation enhances the growth of PSB in dairy wastewater.
Significance and Impact of this Study:  Because PSB utilize H₂S and volatile organic acids (VOA) as an electron source for photosynthesis, and VOA and alcohols as a carbon source for growth, the increase in these bacteria should reduce H₂S, volatile organic compounds and alcohol emissions from the lagoons, enhancing the air quality in dairy farming areas.     

Production of butanol from glucose and xylose with immobilized cells of Clostridium acetobutylicum

Pretreated cotton towels were used as carriers to immobilize Clostridium acetobutylicum CGMCC 5234 cells for butanol or ABE production from glucose and xylose. Results showed that cell immobilization was a promising method to increase butanol concentration, yield and productivity regardless of the sugar sources compared with cell suspension. In this study, a high butanol concentration of 10.02 g/L with a yield of 0.20 g/g was obtained from 60 g/L xylose with 9.9 g/L residual xylose using immobilized cells compared with 8.48 g/L butanol and a yield of 0.141 g/g with 20.2 g/L residual xylose from 60 g/L xylose using suspended cells. In mixed-sugar fermentation (30 g/L glucose plus 30 g/L xylose), the immobilized cultures produced 11.1 g/L butanol with a yield of 0.190 g/g, which were 28.3% higher than with suspended cells (8.65 g/L) during which 30 g/L glucose was utilized completely using both immobilized and suspended cells while 3.46 and 13.1 g/L xylose maintained untilized for immobilized and suspended cells, respectively. Based on the results, we speculated that immobilized cells showed enhanced tolerance to butanol toxicity and the cultures preferred glucose to xylose during ABE fermentation. Moreover, the cultures showed obvious difference when grown between high initial concentrations of glucose and those of xylose. Repeated-batch fermentations from glucose with immobilized cells showed better long-term stability than from xylose. At last, the morphologies of free and immobilized cells adsorbed on pretreated cotton towels during the growth cycle were examined by SEM.