产油微藻筛选和鉴定及其产油性能的研究

为了筛选具有产油能力的微藻,从自然界水体中分离出14株微藻,根据形态特点对它们进行了初步鉴定。对其中12株微藻在自养和异养条件下的生长特性和产油性能进行了比较。通过微藻的生长曲线,生物量和油脂含量等指标,从中筛选出高产藻株并对该藻株进行了分子生物学鉴定。结果表明:藻株Y06在12种微藻中的油脂产量和产率最高,经18S rDNA鉴定确定为栅藻(Scenedesmus abundans)。藻株Y06在自养条件下的油脂产率为9.40 mg/(L.d),在异养条件下的油脂产率为201.29 mg/(L.d)。     

三种微藻对城市二级出水脱氮除磷作用的研究

利用污水培养微藻可去除废水中的氮、磷污染物, 又可以实现生物质生产的耦合。以城市二级出水做为培养基, 选取了小球藻(Chlorella sp.)、栅藻( Scenedesmus sp.)、螺旋藻(Spirulina sp.)为实验藻种, 考察3种微藻的生长特性和脱氮除磷的能力。结果表明, 所选的藻种在模拟二级出水中都能较好生长, 其中栅藻的生物量高于小球藻和螺旋藻, 最大生物量分别为0.328 g·L-1、0.264 g·L-1、0.192 g·L-1, 比生长速率为0.226 d-1、0.213 d-1、0.197 d-1。栅藻的油脂产量达97.35 mg·L-1, 高于其他两种微藻。3种微藻对模拟二级出水都具有较好的脱氮除磷效能, TP的去除率达到90%以上, TN的去除率达到80%以上。实验结果表明利用3种微藻均可达到对二级出水的深度脱氮除磷和生物质的富集。     

一株新型自絮凝凯式拟小球藻的两步培养产油技术

以分离获得的一株新型自絮凝凯式拟小球藻(Parachlorella kessleri) F01为材料, 自养单步培养法为对照, 设计两步培养法, 研究阶段Ⅰ添加葡萄糖兼养和阶段Ⅱ营养元素限制处理对藻细胞油脂积累及絮凝性能的影响。分别采用血球板计数法、干重法、脂染色法测定藻细胞浓度、生物量和总脂含量, 三维荧光光谱分析藻细胞胞外聚合物(Extracellular polymeric substances, EPS)组分和含量。结果表明: (1)两步培养法阶段Ⅰ兼养培养最适葡萄糖浓度为10 g/L, 10d收获时藻细胞油脂产率204.25 mg/(L·d), 是对照组的16.20倍; 静置12h的藻细胞自絮凝率96.1%, 与对照组差异不显著。(2)在阶段Ⅰ基础上, 阶段Ⅱ进行不同元素限制处理培养1d, 低糖组和低糖低氮协同处理组的藻细胞油脂产率分别为242.64和227.61 mg/(L·d), 分别比阶段Ⅰ增加18.8%和11.4%; 培养4d, 低糖、无糖、低氮和低糖低氮协同4种处理组油脂产量显著高于对照组和阶段Ⅰ, 其中低糖低氮协同处理组最高, 达到3.08 g/L, 是对照组的23.69倍, 比阶段Ⅰ增加了51.0%, 而且阶段Ⅱ中4种处理组藻细胞的自絮凝率基本在85.0%以上, 能满足收获要求。(3) F01藻细胞EPS中蛋白类色氨酸物质含量高低与藻细胞自絮凝率大小密切相关, 不同培养处理通过改变藻细胞EPS中蛋白类色氨酸物质的含量而影响其絮凝性能。自絮凝凯式拟小球藻F01是生物柴油生产的优良潜力藻种, 两步培养法能大幅提升其产油效益。产油微藻的自絮凝优势与两步培养法结合, 有望成为解决微藻生物柴油生产技术瓶颈的关键突破口。     

基于微藻培养处理畜禽养殖废水的研究进展

当前规模化畜禽养殖业排放含有大量氮磷、重金属和有机污染物的粪污废水,导致生态环境遭受严重的污染,治理畜禽废水的任务迫在眉睫。由于传统畜禽废水处理方式及应用存在较多不足,基于微藻生物技术处理废水的研究得到越来越多的关注。微藻是一种广泛存在于水体中的单细胞生物,具有高效的脱氮除磷及纳污能力,其主要利用同化作用吸附污水中的氮,通过磷酸化作用吸附、沉降磷,依靠细胞膜上的官能团对重金属进行富集。基于以上生理基础,大多数微藻的氮磷吸附率和重金属富集率可以高达80%。目前微藻对畜禽废水污染组分的处理的研究主要集中在氮磷、重金属,实际应用方式多为高效藻类塘、活性藻、固定化技术、光生物反应器等。但是微藻处理畜禽废水仍存在分子机理研究较少,生产实际经验不足等问题。基于微藻处理畜禽废水的机理,通过综述若干微藻去除氮磷、重金属等污染物的效率,总结国内外微藻废水处理技术的研究及存在问题,展望了微藻废水工程发展前景。     

温度和氮磷浓度对平裂藻和栅藻生长及竞争的影响

为了解温度及氮磷浓度对平裂藻和栅藻生长及竞争的影响,分别在18、25和35℃条件下,以BG11培养基为基础,设置了不同的氮磷浓度组,进行了2种藻的单独培养试验和混合培养试验,结果表明:在纯培养条件下,低营养浓度组(磷浓度分别为0.1和0.5 mg/L)平裂藻在3种温度下基本停止生长,高营养盐浓度组(磷浓度分别为1.0和1.5 mg/L)除18℃1.0 mg/L组停止生长外,其余均能正常生长增殖,且细胞最大密度表现为25℃ 18℃ 35℃, 1.5和1.0 mg/L浓度组细胞生长差异不显著。栅藻在纯培养条件下,细胞密度总体表现为随着营养盐浓度升高而增加的趋势,细胞最大密度表现为18℃ 25℃ 35℃。在混合培养试验中,除35℃、1.5 mg/L组平裂藻对栅藻的竞争抑制参数大于栅藻对平裂藻的竞争抑制参数外,其余试验组均表现为栅藻受平裂藻影响较小。在混合培养体系中, 0.1 mg/L组平裂藻仍基本停止生长, 0.5 mg/L组受栅藻的刺激作用,生长优于纯培养体系。1.0和1.5 mg/L组均受栅藻的抑制作用,且浓度越高抑制作用越强。       

铜绿微囊藻、斜生栅藻对氮磷饥饿的耐受能力研究

在太湖微囊藻水华暴发期间 , 监测了梅梁湾水体中叶绿素 a 和氮磷营养盐的动态变化, 并通过批量培养实验研究了氮磷缺乏对铜绿微囊藻和斜生栅藻生长的影响 , 以探讨两种藻对氮磷饥饿的耐受能力对微囊藻水华形成的影响。结果表明 , 在水华期间 , 水体的氮磷营养盐浓度并不高, 并处于波动状态; 铜绿微囊藻和斜生栅藻对氮磷饥饿的耐受能力存在很大差异 : 在无磷培养基中 , 铜绿微囊藻能保持 8 天的指数生长, 而斜生栅藻在无氮或无磷培养基中生长都非常缓慢。两种藻对磷饥饿的耐受能力均高于氮。通过生长速率的比较, 发现铜绿微囊藻对磷饥饿的耐受能力明显高于斜生栅藻, 因此更能够适应波动的营养供给条件。对营养盐饥饿耐受能力的不同导致两种藻在营养盐波动的情况下微囊藻更易取得优势, 成为水华期间浮游植物的优势种。     

废水培养微藻藻种遴选及其组分积累的研究进展

利用废水培养微藻能够降低微藻的培养成本,同时削减污染.微藻的蛋白质、多糖和油脂等组分是影响其后续资源化利用的重要因素.本文重点综述了以废水为基质培养微藻的研究进展,从组分积累的角度,分析了微藻种类的选择依据,探讨了影响微藻生长的因素和提高产量的方法,并对藻体中组分的合成机制进行了讨论,提出未来废水培养微藻技术面临的挑战...     

不同氮浓度对一株产油绿球藻生长、脂类积累及脂肪酸分布的影响

富含多不饱和脂肪酸的产油微藻是开发高值微藻油的理想原料,一株产油微藻是否具有高值油脂开发潜力需要评估其油脂产量、中性脂比例与多不饱和脂肪酸分布等指标。低氮胁迫是研究微藻油脂积累理想的方式之一,以一株产油绿球藻(Chlorococcum sp.)为实验材料,以硝酸钠(NaNO_3)为氮源,设置17.6 mmol/L、5.9 mmol/L和3.5 mmol/L三种氮浓度,跟踪测定产油绿球藻生长、脂类组成及多不饱和脂肪酸分布的时相变化。结果显示,3.5 mmol/L和5.9 mmol/L氮浓度条件下,产油绿球藻取得了2.55 g/L和2.51 g/L的总脂产量,远高于17.6 mmol/L氮浓度组的总脂产量(1.43 g/L);降低氮浓度可以提高中性脂比例,3.5mmol/L氮浓度组取得最高的中性脂比例,为88.6%总脂质(Total lipid,TL),高于5.9 mmol/L氮浓度组(86.3%TL)和17.6 mmol/L氮浓度组(80.5%TL);降低氮浓度可以改变产油绿球藻的脂肪酸在不同脂类分子中的分布,促进脂肪酸更多分布于中性脂中,其中,3.5 m mol/L TL氮浓度组培养结束时,α-亚麻酸在中性脂的比例由接种时的33.9%提高到86.5%。适宜氮浓度对于提高产油绿球藻总脂产量、中性脂比例而多不饱和脂肪酸分布具有重要作用,产油绿球藻积累的α-亚麻酸在低氮条件下更倾向于分布在中性脂中,是一株具有高值化微藻油开发价值的藻株。     

斜生栅藻在不同磷条件下氮生态幅的研究

为获得斜生栅藻（Scendesmus obliquus）的氮生态幅,研究根据中华人民共和国地表水环境质量标准磷浓度界定,利用谢尔福德（Shelford）耐受定律进行曲线拟合对斜生栅藻在低磷（0.02 mg/L）、中磷（0.2 mg/L）和高磷（0.4 mg/L）三种不同磷浓度下氮的生态幅进行定量表达,获得三种磷起始条件下斜生栅藻生长的最佳氮浓度、氮适宜生长范围和氮耐受范围。研究表明,在三种磷条件下斜生栅藻生长的最佳氮浓度分别为1.02、8.91和18.05 mg/L,对应的最大比生长速率分别为（0.1420.006）、（0.3140.002）和（0.3460.007） /d,氮适宜生长范围分别为（0.521.52）、（4.4813.34）和（11.7224.38） mg/L,氮耐受限度分别为（0.022.02）、（0.0517.77）和（5.3930.71） mg/L。这表明富营养化水体可能引起斜生栅藻的大量生长、繁殖,也暗示了斜生栅藻能作为高氮水环境的一个良好指示生物。       

小型生活污水受纳水体浮游植物增长的氮、磷限制研究

为研究生活污水处理后其受纳水体中浮游植物增长的氮磷限制,选取某生活污水处理系统的受纳水体为研究对象,依据我国《城镇污水处理厂污染物排放标准》(GB189182002)一级A标准(氨氮5 mg/L和磷0.5 mg/L)进行氮磷营养盐最高浓度和浓度梯度添加微宇宙实验模拟实验。最高浓度添加实验结果显示N、P双添加的实验组中3d后叶绿素a的浓度显著(P0.05)高于单独添加氮和单独添加磷实验组。因此,氮和磷是被研究水体浮游植物生长的共同限制因子。同时结果还暗示受纳水体接纳处理后的生活污水仍可能会造成浮游植物在短期内剧烈增长。浓度梯度添加实验结果显示,将磷控制在0.27 mg/L或者将氮控制在1.0 mg/L以下,可以有效降低被研究水体浮游植物的增长。据此可以进一步严格生活污水处理后的排放标准以降低受纳水体水华的风险。     

Effect of nitrogen and phosphorus concentration on their removal kinetic in treated urban wastewater by Chlorella vulgaris

This study evaluates the feasibility of removing nutrients by the microalgae Chlorella vulgaris, using urban wastewater as culture medium, namely the effluent subjected to secondary biological treatment in a wastewater treatment plant (WWTP). For this, laboratory experiments were performed in batch cultures to study the effect of initial nitrogen and phosphorus concentrations on growth and reduction of nutrient performance of C. vulgaris. The microalga was cultivated in enriched wastewater containing different phosphorus (1.3-143.5 mg x L(-1) P.PO4(3-)), ammonium (5.8-226.8 mg x L(-1) N-NH4+) and nitrate (1.5-198.3 mg x L(-1) N-NO3-) concentrations. The nutrient removal and growth kinetics have been studied: maximum productivity of 0.95 g SS x L(-1) x day(-1), minimum yield factor for cells on substrate (Y) of 11.51 g cells x g nitrogen(-1) and 0.04 g cells x g phosphorus(-1) were observed. The results suggested that C. vulgaris has a high potential to reduce nutrients in secondary WWTP effluents.     

利用市政污水培养Chlorella vulgaris生产生物柴油

为了考察利用南昌市政污水规模化培养富油微藻生产生物柴油,同时达到净化污水的目的,取南昌市青山湖污水处理厂未经任何处理的市政污水作为普通小球藻(Chlorella vulgaris)生长的培养液。监测了C.vulgaris在市政污水中连续培养10 d的特定生长率、生物质产量以及与之相关的市政污水中氨氮(NH4+-N)、总磷(TP)、化学需氧量(COD)、总悬浮固体(TSS)和挥发性悬浮固体(VSS)的清除情况。实验表明:营养物质的水平显著地影响了C.vulgaris的生长。C.vulgaris的生长率在培养8 d后达到最大,OD680为2.856,总的生物质产量日均最大积累速率为0.01 g/L,油脂含量为干质量的18%,油脂的平均日产量为0.001 g/L。培养10 d内NH4+-N、TP和COD的去除率分别为50.0%、32.1%和26.0%,TSS和VSS的日平均去除速率分别为0.01 g/L和0.006 1 g/L。     

富营养废水中碳氮磷浓度对布朗葡萄藻总脂和总烃的影响

以经过二次过滤的富营养化鱼塘养殖污水为培养液,添加外源的碳、氮、磷元索,研究了污水中不同的外源无机碳、总氮和总磷浓度对布朗葡萄藻(Botryococcus braunii)生物量、总脂和总烃含量的影响.结果表明:(1)以NaHCO3作为碳源,布朗葡萄藻的生物量和总脂含量在外源无机碳浓度为5～10 mg/L时最高,总烃含量在外源无机碳浓度为15mg/L时最高.(2)以KNO3作为氮源,布朗葡萄藻的生物量在总氮浓度为15mg/L时最高,总脂含量在总氮浓度为2mg/L时最高,总烃含量在总氮浓度为20mg/L时最高.(3)以KH2 PO4作为磷源,布朗葡萄藻生物量在总磷浓度为2mg/L时最高,总脂含量和总烃含量在总磷浓度为1.5 mng/L时最高.     

On the nitrogen and phosphorus removal in algal photobioreactors

This study shows results of nitrogen and phosphorus removal by microalgae (tertiary treatment) in a prototype of tubular photobioreactor tested under controlled and uncontrolled conditions. The wastewater was the supernatant coming from a secondary settler of a municipal wastewater activated sludge treatment plant without nitrification and denitrification units. The algal biomass was directly selected from the supernatant and it was principally composed of genus Scenedesmus (autochthonous algae). All the experiments evaluated both nitrogen and phosphorus removal and biomass and lipid production. A satisfactory nutrients removal - about 99.9% for the nitrogen and phosphorus - and a specific biomass productivity of 0.25 g/l d have been obtained in the indoor photobioreactor; less satisfactory results have been reached in the outdoor photobioreactor because of ambient condition instability and limiting nutrients concentration.     

Removal of nitrogen and phosphorus from wastewater using microalgae immobilized on twin layers: an experimental study

Removal of nitrogen and phosphorus from wastewater by two green microalgae (Chlorella vulgaris and Scenedesmus rubescens) was investigated using a novel method of algal cell immobilization, the twin-layer system. In the twin-layer system, microalgae are immobilized by self-adhesion on a wet, microporous, ultrathin substrate (the substrate layer). Subtending the substrate layer, a second layer, consisting of a macroporous fibrous tissue (the source layer), provides the growth medium. Twin-layers effectively separate microalgae from the bulk of their growth medium, yet allow diffusion of nutrients. In the twin-layer system, algae remain 100% immobilized, which compares favourably with gel entrapment methods for cell immobilization. Both microalgae removed nitrate efficiently from municipal wastewater. Using secondary, synthetic wastewater, the two algae also removed phosphate, ammonium and nitrate to less than 10% of their initial concentration within 9 days. It is concluded that immobilization of C. vulgaris and S. rubescens on twin-layers is an effective means to reduce nitrogen and phosphorus levels in wastewater.     

Phycoremediation of dairy and winery wastewater using <Emphasis Type="Italic">Diplosphaera</Emphasis> sp. MM1

A new green microalgal species was isolated, identified and investigated for its biomass production and nutrient removal efficiency in dairy and winery wastewater in this study. The 18S rRNA-based phylogenetic analysis revealed that this new strain is a Diplosphaera sp. and was designated strain MM1. The growth of this strain was evaluated in different diluted dairy and winery wastewaters. The highest algal biomass production (up to 2.3 g L^?1) was obtained in dairy wastewater (D3; dairy wastewater 1:2 deionised water) after 14 days of culture. However, for winery wastewater, the highest algal biomass production (up to 1.46 g L^?1) was obtained in wastewater combination W2 (winery wastewater 1:1 deionised water) after 14 days of culture. Turbid dairy wastewater with high concentration of nitrogen and phosphorous slowed down the initial growth of the alga. However, at the end of day 14, biomass production was nearly twofold higher than that of winery wastewater. The findings from both types of wastewater suggest that Diplosphaera sp. MM1 has potential for its application in generating biomass with simultaneous remediation of nutrient-rich wastewater.     

钝顶螺旋藻SP1(Spirulina platensis)对集约化养殖尾水氮磷的去除效果

针对集约化养殖模式后期硝酸盐氮和磷酸盐浓度较高的问题, 实验设置生物絮团养殖尾水(BFW)和BG11培养液(BGW)两种水体环境, 并以池塘常见优势微藻--绿色颤藻OC1(Oscillatoria chlorina)作为对比, 研究分析了钝顶螺旋藻SP1(Spirulina platensis)对集约化养殖尾水氮磷的去除效果及其生长状况。结果发现, 在BFW组中两种微藻均对硝酸盐氮(NO3--N)、总无机氮(TIN)和磷酸盐(PO43--P)去除效果明显(P<0.05), 其中, 螺旋藻对NO3--N、TIN和PO43--P的最大去除率分别为79.60%、46.06%和98.55%, 相应的浓度值分别从130.04 mg·L-1、130.85 mg·L-1和10.23 mg·L-1降至26.53 mg·L-1、70.58 mg·L-1和0.15 mg·L-1,其数量降低的绝对值分别为103.51 mg·L-1、60.27 mg·L-1、10.08 mg·L-1; 在BGW组中两种藻对氮磷均具有一定的去除效果, 但总体仍低于BFW组。实验过程中两种微藻的细胞数量均无明显变化(P>0.05)。可见, 钝顶螺旋藻SP1和绿色颤藻OC1均可在BFW和BGW两种水体营养环境下存活, 且对水中的氮磷均有良好的去除效果; 虽然颤藻亦是集约化养殖水环境中的常见微藻优势种, 但它能分泌蓝藻毒素, 因此, 从产业应用的可行性考虑可将螺旋藻作为集约化养殖尾水净化的备选藻株。     

Wastewater treatment at the Houghton Lake wetland: Hydrology and water quality

Lagoon-treated wastewater was discharged to a natural peatland to remove nutrients. For thirty consecutive years, an average of 600,000 m³ of treated water was discharged to the Porter Ranch peatland near the community of Houghton Lake, Michigan. This discharge was seasonal, commencing no sooner than May 1 and ending no later than October 31. During the winter half-year, treated wastewater was stored at the lagoon site. This water contained 3.5 mg/L of total phosphorus, and 7 mg/L of dissolved inorganic nitrogen. Other wastewater quality parameters were CBOD₅ = 15 mg/L, TSS = 34 mg/L, and fecal coliforms at 66 cfu/100 ml. The peatland was large, about 700 ha, but the zone that provided wastewater polishing was approximately 100 ha. Outflows from the larger peatland showed no effects of the discharge, and maintained concentrations of 40 μg/L of phosphorus, and 85 μg/L of ammonia nitrogen. Nutrients were stored in the 100-ha irrigation area, which removed 94% of the phosphorus (53 metric tons) and 95% of the dissolved inorganic nitrogen. All other constituents were also removed in the irrigation area, except for pass-through substances such as chloride. Phosphorus was stored in new biomass, increased soil sorption, and accretion of new soils and sediments, the last being dominant. A simple growth and uptake model described the removal of phosphorus, with an uptake rate coefficient that did not change over time. Thus, rates in this system were stable over time, and the P-removal capacity did not diminish. The irrigation area underwent large changes in ecosystem structure. There was an initial fertilizer response, characterized by much larger standing crops of vegetation. There was also a plant community shift, from the initial sedge-willow cover type to a cattail-dominant cover type. This new cattail patch became a floating mat.