谷氨酸脱盐废液培养普通小球藻(Chlorella vugaris)C9-JN2010

研究小球藻在谷氨酸脱盐废液中生长,藻细胞组分及废液中营养利用的效果。结果表明:停留时间10 d,小球藻在体积分数为2%脱盐废液中生长最佳,对总糖、总氮及总磷的消耗率分别为99.1%、96.0%和97.0%;细胞干质量、比生长速率和最大生产强度分别为5.60 g/L、0.223 d-1和560.0 mg/(L.d),细胞中粗蛋白质、粗脂肪、粗纤维、粗灰分及总磷含量(以质量百分数计)分别为50.0%±5.0%、10.0%±2.0%、4.0%±0.5%、6.0%±0.5%和0.80%±0.10%。     

沼液中化学成分的研究

通过用硅胶、凝胶柱层析和制备液相纯化,从沼液浓缩物里分离出四个化合物,经鉴定其中一个为新化合物,将其命名为24-降齐墩果烷-4,12-二烯-3,22-二酮(1),其余三个为已知化合物,8-羟基-3,4-二氢喹啉-2酮(2)、(E)-3-(2-羟基苯)丙烯酰胺(3)、苯乙酸(4)。这些化合物均是首次从鸡粪沼液中分离出来。     

不同氮磷浓度下周丛生物对水体中磺胺和恩诺沙星的去除

周丛生物膜是一种对水体污染物净化的新兴生物技术。有关水体不同氮磷营养水平下周丛生物对水体抗生素类污染物去除作用的研究还未见报道。本研究设置4个氮磷营养盐浓度水平[N-P (mg·L^-1):2-0.2、5-0.5、8-0.8、11-1.1],用塑料筐装置在室外培养周丛生物膜,对其生长、光合活力、物种组成以及对磺胺和恩诺沙星去除作用进行中型模拟试验。结果表明: 各处理组中周丛生物的生物量随培养时间的增加而升高,光合色素含量和光合活力则呈现先降低后上升的“单峰”模式,表明生物膜中的藻类会受到抗生素的胁迫,但可快速适应并恢复活力。除此之外,不同氮磷浓度处理造成各组生物群落组成差异,随营养盐浓度的升高,周丛藻类物种丰富度逐渐下降,但各处理胶网藻和小球藻都具有较高的相对丰度;16S rRNA高通量测序发现,根瘤菌科、放线菌门和莫拉氏菌科菌群在(N-P)2-0.2组显著富集,而几丁质嗜菌科在4个处理中的相对丰度都处在最高水平。所有处理的磺胺去除率均高于50%,而恩诺沙星去除率均达到90%以上,其中,(N-P)2-0.2 mg·L^-1组对磺胺的去除率(65.8%)显著高于其他各组,但各处理对恩诺沙星的去除率差异不显著,表明周丛生物在较宽的N-P营养水平范围内对磺胺和恩诺沙星均具有良好的去除能力。各处理组对水体可溶性氮的去除效果不明显,但对可溶性磷的去除效果显著。本研究为水体磺胺和恩诺沙星的生态去除提供了基础数据,为研发水体抗生素类新型污染物生态去除技术提供了新思路。     

小球藻去除水产加工废水中氨态氮的初步研究

目的：确定小球藻去除水产行业加工废水中氨态氮的优化环境条件,为利用小球藻处理此类废水的规模化发展提供基础的实验数据。方法：在静止摇瓶培养方式下,逐一改变废水与小球藻培养液的混合体系的初始pH、温度或光照强度等条件,定时测定体系的氨态氮含量,并根据其随培养时间的变化确定优化环境条件。结果：当小球藻藻液的初始浓度为1×10~6个/mL左右,且小球藻培养液与水产加工废水的配比为3：2时,小球藻去除氨态氮的优化环境条件为：初始pH 7．5、培养温度25℃、光照时间12h/d,光强5300k,此时氨态氮的去除率达到77%。结论：实验条件下小球藻能够较好地去除水产加工废水中的氨态氮,为小球藻规模化处理此类废水提供了基础数据。     

典型农田土壤中丛枝菌根真菌-根际细菌互作及与氮磷利用的关系

在农业生态系统中,土壤微生物是土壤-作物系统养分循环的重要驱动力,其中丛枝菌根真菌(Arbuscularmycorrhizalfungi,AMF)能够促进作物对养分的吸收,适应逆境胁迫。【目的】进一步揭示AMF和根际细菌群落的跨界网络互作,挖掘与作物氮磷利用显著相关的关键微生物类群,揭示关键类群的生态网络特征。【方法】利用Illumina测序技术对3种典型农田旱地土壤(黑土、潮土和红壤)中AMF和根际细菌群落结构进行分析;构建互作网络并利用偏冗余分析、相关性分析探究了与氮磷利用相关的潜在关键类群。【结果】3种土壤中AMF与根际细菌均以正相互作用为主。不同土壤中AMF与根际细菌互作关系差异明显,在红壤中跨界互作最为密切,其中球囊霉属真菌(Glomus)与根际细菌中的放线菌(Actinobacteria)和变形菌(Proteobacteria)之间的交互作用最多。而在黑土中主要体现为根际细菌的界内互作。与氮磷利用率显著相关的关键微生物类群主要属于球囊霉属真菌、放线菌和α变形菌。【结论】典型旱地土壤中AMF与根际细菌的正相互关系对作物氮磷利用有潜在促进作用,关键类群在有机质和养分贫乏的红壤中可能起到更重要的作用。     

典型抗生素在土壤-水-蔬菜系统中迁移分布的研究

通过施用畜禽废物和抗生素污染进行芥菜盆栽试验,从而研究典型抗生素(QNs、SAs和TCs)在土壤.水一蔬菜系统的迁移分布规律.研究结果表明:蔬菜、渗漏水、土壤中均检测到多类抗生素,抗生素在不同介质中的含量与施加抗生素的种类及其含量等因素有关.盆栽后,土壤中抗生素残留量为:QNsTCsSAs,施加猪粪总体上不会造成土壤抗生素污染(ENF除外);只施用畜禽废物的土壤渗漏水中抗生素含量较低(<40.77 μg·L-1),而添加抗生素污染的土壤渗漏水中抗生素含量明显升高(<2.28 mg·L-1),施加猪粪会造成渗漏水抗生素污染;SMT、ENF、OTC、TC这四种药物在盆栽芥菜中均被检出,其中OTC检出率最高,而SMZ则未检出,施加猪粪会造成芥菜抗生素污染.     

脱氮功能菌去除市政废水中氮素的研究

为了探讨实验室筛选获得的氨氧化细菌CM-NRO14和反硝化细菌CM-NRD3联合去除市政废水中氮素的应用价值,采用了两级A/O工艺进行菌株去除废水中氮素的小试实验,最后将菌株用于废水脱氮工程中。结果表明,脱氮功能菌实现了短程硝化-反硝化,氨氮去除率在98%以上,总氮去除率在75%以上, COD (化学需氧量)去除率大于90%,出水各项指标均低于城镇污水处理厂污染物排放一级(A)标准。脱氮功能菌在去除市政废水中氮素方面有很高的应用价值,可用于城镇污水处理厂提标改造等。     

不同来源小球藻对岩溶水Ca2+、HCO3-利用的初步研究

以外源小球藻和岩溶区筛选出的土著小球藻为研究对象, 在封闭体系中比较研究了两种不同来源小球藻对典型岩溶水中Ca2+、HCO3-的利用、藻细胞数量与其对Ca2+、HCO3-的利用率的关系和体系pH的变化。结果表明, 土著小球藻利用Ca2+、HCO3-的能力强于外源小球藻, 但外源小球藻对Ca2+的利用量高于土著小球藻, 而二者对HCO3-的利用量相同, 并且外源小球藻能够以胞外CaCO3形式产生沉淀, 而土著藻则不能形成沉淀。其次两体系中pH的变化显示, 两种小球藻光合作用都是先以水体中CO2为光合作用碳源, 然后利用HCO3-。外源小球藻能将岩溶水中29.648%的HCO3-吸收, 而土著藻能将40.625%的HCO3-通过其在食物链中的初级生产地位将岩溶碳汇转化进入到生态系统, 表现为净碳汇效应。       

利用基于CRISPR-Cas9 的基因编辑技术对miR-29a在GT1-7 细胞中进 行基因敲除

目的:本文利用CRISPR-Cas9技术在GT1-7细胞中对miR-29a基因进行基因编辑,用于构建miR-29a基因敲除GT1-7细胞模型。方法:通过构建Cas9稳转的GT1-7细胞株并转染sgRNA质粒用于在靶向miR-29a基因区域引发突变。然后构建EGFP与sgRNA共表达质粒并转染Cas9稳转GT1-7细胞,利用流式细胞仪富集表达绿色荧光蛋白的阳性细胞和分选阳性单克隆细胞。最后利用实时荧光定量PCR(realtimefluorescencequantitativePCR)对富集细胞和单克隆细胞进行miR-29a表达量检测。结果:T7E1检测结果显示CRISPR-Cas9系统有效地在miR-29a基因区域引发了突变。荧光定量PCR结果显示,与对照组相比,富集后阳性细胞miR-29a的表达量整体下降了50%左右(P0.05)。此外,通过流式筛选获得了一个纯合miR-29a基因敲除细胞克隆,与对照组相比,其miR-29a的表达量下降了75%左右(P0.05)。结论:本文建立了一种有效编辑GT1-7细胞基因的方法,并采用该方法构建了miR-29a稳定敲除细胞模型。     

LC-MS/MS法检测人头发中利培酮及其代谢物9-羟利培酮含量的实验研究

目的:建立高效液相色谱-三重四级杆质谱联用(LC-MS/MS)法检测人头发中利培酮(RIP)及其代谢物9-羟利培酮(9-OH-RIP)含量的方法。方法:采用同位素内标氘4-利培酮(RIP-d4)及氘4-9-羟利培酮(9-OH-RIP-d4),流动相A为10 mmol/L醋酸胺溶液(甲酸调p H值为4.0),流动相B为乙腈,A/B=70/30,流速为0.3 m L/min,等度洗脱4.00 min。色谱柱为安捷伦Zorbax SB C18(2.1×50 mm,1.8μm),柱温30℃。准确称取20 mg丙酮清洗过晾干剪碎成粉末的头发样本,加1N氢氧化钠(Na OH)超声2 h,等量酸中和后,加200μL1N氢氧化钠溶液及5.0 m L的甲基叔丁基醚(MTBE)提取涡旋1分钟,3000×g离心5 min后取上清液在40度水浴下氮气吹干后用100μL流动相复溶,进样2μL经LC-MS/MS检测。MRM监测离子对:RIP:m/z 411.2→191.0,9-OH-RIP:m/z 427.2→207.1,RIP-d4:m/z 415.2→195.2,9-OH-RIP-d4:m/z 431.2→211.1。结果:利培酮及9-羟利培酮线性范围分别为0.5-25 ng/mg,0.0025-0.15 ng/mg,提取回收率均70.0%,方法回收率均在85.0%-115.0%之间,线性r均0.999,精密度和重现性RSD均15%。结论:本研究建立了采用LC-MS/MS法检测人头发中利培酮及9-羟利培酮含量的方法,该法快速、简单、准确、重现性好。     

普通小球藻对养殖污水脱氮除磷的效果研究

随着我国养殖业的不断发展,养殖污水排放量的日益增加,养殖污水的高氮、磷含量导致水体富营养化问题日趋严重。小球藻是光能自养生物,能有效同化氮、磷,使污水中的氮、磷减少。本研究通过在实验室模拟不同氮、磷含量的养殖污水环境,分析小球藻对氮、磷的去除效果;在此基础上,用小球藻处理某养殖场污水;并联合膨润土与小球藻,探究两者脱氮除磷的协同作用能力及膨润土对小球藻细胞沉降的效果。结果表明,小球藻对模拟污水的氨氮去除率可达80%,对磷酸根的最高去除率接近100%;对养殖污水中的氮、磷也有一定的去除效果;但养殖污水成分复杂,小球藻的生长被抑制。膨润土与小球藻的结合,能够提高污水中的氮磷去除率并帮助藻细胞快速沉降,为污水处理后藻细胞的收集处理提供了有效方法。     

半连续及连续培养小球藻减排沼液及CO2

采用半连续或连续模式培养小球藻,考察小球藻减排沼液和CO2的能力。结果表明:在半连续培养模式中,当更新率为30%时,沼液中的N、P质量浓度可分别稳定在16～18和0.4～0.6 mg/L,达到污水二级排放标准;提高更新率到40%以上,3 d后微藻生物量及其对沼液中N、P的吸收达到动态平衡,但N、P去除率未达到污水直接排放标准;在连续培养模式中,分别选用20%及30%的日更新率,7 L规模12 d后沼液中的总氮(TN)仍高达55.64 mg/L。说明大规模培养条件下的光限制是微藻法减排沼液的主要制约因素。     

pH值对沼液培养的普通小球藻生长及油含量积累的影响

以50％的沼液为普通小球藻的全营养培养基,考察培养基的起始pH值对小球藻生长及油脂含量的影响,普通小球藻对不同初始pH的沼液中氮、磷的去除情况。设定了2组实验,一组只调节初始接种培养液的pH,分别为6.0、6.5、7.0、7.5、8.0、8.5;另一组将培养液pH分别固定在6.0、6.5、7.0、7.5、8.0、8.5,pH用稀HCl和NaOH进行调节。研究发现在pH 6.5和pH 7.0的偏酸环境有利于小球藻生长,而pH在7.0~8.5的偏碱性条件下有利于小球藻油脂的积累,因此综合小球藻生长和油脂积累2个因素,得到最适合小球藻生长和油脂积累的pH为7.0。培养结束后沼液中氮磷的去除率分别达到了95％和97％,沼液中的总氮由原来的134.91 mg/L降至4.86 mg/L,总磷由10.19 mg/L降到0.32 mg/L。     

菌藻共生系统对序批式反应器处理养猪废水脱氮除磷效果及微生物群落结构的影响

[背景]养猪废水作为高浓度有机废水,是导致我国农业面源污染的主要因素之一.目前采用菌藻共生系统处理养猪废水越来越受到关注,与传统序批式反应器(Sequencing Batch Reactor,SBR)相比,藻辅助SBR具有提高脱氮除磷效果、增加污泥活性和降低能源消耗的特点.[目的]针对SBR中菌藻共生系统对养猪废水脱氮...     

三种不同小球藻去除亚硝态氮和氨氮能力的研究

为了研究小球藻在不同质量浓度的氨氮和亚硝态氮环境下的去除能力与生长效果,从实验室挑选3株不同的小球藻CV315-1(Chlorella sp.)、CV315-2(Chlorella sorokiniana)和CV315-3(Chlorella pyrenoidosa)分别置于不同质量浓度的氨氮和亚硝态氮模拟污水中,在温...     

Operation and modeling of bench-scale SBR for simultaneous removal of nitrogen and phosphorus using real wastewater

Experimental work was carried out on nitrogen and phosphorus removal from real wastewater using a bench-scale SBR process. The phosphorus removal was stable and the phosphorus concentration remaining in the reactor was maintained within 1.5 ppm, regard-less of the addition of an external carbon source. In the case of nitrogen, an external carbon source was necessary for denitrification. The effect on denitrification with the addition of various carbon sources, such as glucose, methanol, acetate, and propionate, was also investigated. Acetate was found to be the most effective among those tested in this study. When 100 ppm (theoretical oxygen demand) of sodium acetate was added, the average rate of denitrifiaction was 2.73 mg NO₃⁻-N (g MLSS)⁻¹ h⁻¹, which wasca. 4 times higher than that with the addition of 200 ppm of methanol. The phosphorus and nitrogen concentrations were both maintained within 1.5 ppm by the addition of an appropriate amount of a carbon source during a long-term operation of the SBR. The mathematical modeling was performed using Monod kinetics, other microbial kinetics, mass balances, and stoichiometry. The modeling was found to be useful for predicting the SBR operation and optimizing the HRT.     

Process hydraulics, distributed bacterial states, and biological phosphorus removal from wastewater

Hydraulic characteristics of biological wastewater treatment systems were shown to affect bacterial state distributions and system performance through mathematical simulations. The term "state" is used here to mean the microbial storage product and biomass content of a bacterium. The traditional approach to simulating biological treatment processes assumes "lumped" (average) states, rather than accounting for variable states across bacterial populations. Distributed states were previously suggested as critical to enhanced biological phosphorus removal (EBPR), but the factors that cause distributed states were not evaluated. A primary driver for distributed state development is variable hydraulic experiences of bacteria as they cycle through completely mixed reactors, and so process characteristics that affect hydraulics were hypothesized to affect state distributions. Two design characteristics affecting system hydraulics were evaluated using a new distributed state simulation program (DisSimulator 1.0): total hydraulic residence time (HRT) and numbers of reactors in series. Distributed predictions consistently predicted worse EBPR performance than did the lumped approach. Increasing HRTs (with constant solids retention times) tended to increase state distributions, to increase the differences between lumped and distributed simulation predictions, and to decrease predicted EBPR performance. As the numbers of reactors in series increased, distributed predictions tended to converge with lumped simulation predictions. Distributed simulations tended to predict a greater benefit to using reactors in series than did lumped simulations. This work provides guidance for new strategies to improve EBPR by minimizing state distributions. The targeted hydraulic characteristics may be more important to EBPR than previously recognized due to their effects on distributed states.     

氮源对C.vulgaris油脂积累的影响

实验室条件下,考察了在发酵过程中不同氮源对小球藻的生物量和油脂积累的影响,确定了小球藻的最佳氮源;并对比分析了含氮培养与缺氮培养的生物量、油脂含量、氮消耗量、生物量氮消耗比率和油脂氮消耗比率的不同。结果表明:小球藻在1.6 g/L Na NO3时获得最大生物量,为562.2 mg/L,在0.8 g/L Na NO3时获得最大相对油脂含量为12.01%;以油脂含量为考察指标时,培养小球藻的最佳氮源为0.8 g/L Na NO3;缺氮培养时,最大油脂含量为13.49%,比含氮培养高约15%;含氮培养时,最高生物量为626.3 mg/L,比缺氮培养高约1.9倍。氮源对生物量,相对油脂含量,生物量氮消耗比率和油脂氮消耗比率具有明显的影响。藉此,提出了通过改变培养方式,达到调控小球藻细胞内生理代谢组分的可行性。     

Ammonia removal from anaerobically digested dairy manure by struvite precipitation

Ammonia is one of the most important contaminants impairing the quality of water resources. When this is considered along with the fact that the global demand for nitrogenous fertilizers is in constant rise, the need for recovery as well as removal of nitrogen is well justified. Crystallization of N and P in the form of struvite (MgNH₄PO₄·6H₂O), which is a slow releasing and valuable fertilizer, is one possible technique for this purpose. This study investigated the removal of NH₄⁺ through struvite precipitation from the effluents of one- (R1) and two-phase (R2) anaerobic reactors digesting dairy manure. To force the formation of struvite in the anaerobic reactor effluents, Mg²⁺ ion was added by using both Mg(OH)₂ and MgCl₂·6H₂O. To prevent the effect of different total phosphorus (TP) concentration in the effluents of R1 and R2, as well as to not limit the formation of struvite, an excess amount of PO₄³⁻ (0.14 M) was added in the form of Na₂HPO₄. Different stoichiometric Mg²⁺:NH₄⁺:PO₄³⁻ ratios were tested to determine the required Mg²⁺ concentrations for maximum NH₄⁺ removal by keeping NH₄⁺:PO₄³⁻ ratio constant for the effluents of reactors R1 and R2. The results revealed that very high NH₄⁺ removal efficiencies (above 95%) were possible by adding Mg²⁺ ions higher than 0.06 M concentration in the effluents from reactors R1 and R2. It was also observed that the initial pH adjustment to 8.50 using NaOH did not result in any significant increase in the removal of NH₄⁺ and the removal of NH₄⁺ in the reactors treated with MgCl₂·6H₂O was higher than those treated with Mg(OH)₂ for the same Mg²⁺ concentration.