碳源小球藻强化人工湿地处理养猪沼液的效能与机制

针对传统人工湿地处理畜禽养殖沼液反硝化不足、脱氮效果不佳的难题,该研究将小球藻(Chlorella sorokiniana)作为人工湿地外加碳源,构建小球藻强化人工湿地脱氮系统,通过添加不同浓度的小球藻（0、0.25、0.5、1.0 g/L）来研究系统对养猪沼液水质的去除效果及溶解性有机物（DOMs）的变化规律,并从微生物角度解释强化脱氮机制。结果表明：（1）与未添加小球藻的人工湿地相比,总氮的去除率明显增高,最高可达70.24%,硝氮去除率最高达93.45%;(2)基于三维荧光平行因子分析,DOMs中3组分分别为富里酸/类腐殖酸物质、高分子芳香族物质和类腐殖酸物质;小球藻的添加增强了养猪沼液中DOMs的去除,同时降低了类腐殖酸的含量;（3）小球藻的添加明显增加了人工湿地中微生物的Alpha多样性。其中,门水平上,与反硝化作用相关的优势菌门（变形菌门）相对丰度明显增加;属水平上,亚硝化单胞菌属（Nitrosomonas）和湖沉积杆菌属（Limnobacter）在系统氨氧化和脱氮过程中发挥重要作用。该研究为小球藻作为外加碳源增强人工湿地脱氮除磷提供了理论支持。     

污水厂尾水中消毒副产物对小球藻的生长生理影响

通过检测无锡某污水厂疫情期间排放尾水中消毒副产物的种类和浓度,发现一氯乙酸（CAA）占全年总检出量的30.22%。采用毒性标准实验方法研究CAA对小球藻生长和抗氧化系统的影响。结果表明在CAA胁迫下,小球藻生长速率减慢,最大光化学量子产量（Fv/Fm）较空白组出现明显下降,96 h叶绿素 a 浓度明显降低。CAA对小球藻的96 h半数效应浓度（EC₅₀）为14.734 mg·L^-1。中、低浓度CAA胁迫会诱导超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性和还原型谷胱甘肽（GSH）含量的上升,减少丙二醛（MDA）含量的积累;而高浓度胁迫下,SOD、CAT活性和GSH含量持续下降,MDA含量先升高再下降。     

Effect of nutrient conditions on the toxicity of naphthalene to Chlorella pyrenoidosa

The toxicity of naphthalene to a freshwater microalga, Chlorella pyrenoidosa, and the subsequent recovery of algae from the damage were investigated under two nutrient conditions, either enriched with nitrogen (N) and phosphorus (P), or starved of N and P. Results showed that C. pyrenoidosa was more sensitive to naphthalene under N,P-enriched condition, and the inhibitory rate generally increased at first and then decreased gradually with the evaporation of naphthalene under both nutrient conditions. Enriched N, P reduced the inhibitory rate at initial naphthalene concentration of 5 and 10 mg/L, but enhanced it at 100 mg/L, at which more severe ultrastructure damages were found than those under N,P-starved condition. Observed damages included partly or totally disappearance of nucleolus, nuclear, and plasma membranes. According to the chlorophyll content and cell density measurements, C. pyrenoidosa could recover from naphthalene damage with initial concentrations 6 50 mg/L in 7 days under both nutrient conditions, while they could not recover if the initial concentration of naphthalene was at 100 mg/L. Under the N,P-starved condition, the inability of C. pyrenoidosa to recover from the naphthalene damage was consistent with the results of high inhibitory rate, low value of specific growth rate (SGR, 0.05 day??1), and the severe destruction of cell structure. However, under the N,P-enriched conditions, the observed lower inhibitory rate, higher value of SGR (0.55 day??1), and the intact cell structure of most cells suggested that algae could potentially recover from the naphthalene damage.     

氧化乐果对小球藻的毒性研究

以小球藻(Chlorella vulgaris)为实验生物,通过研究不同浓度(0、1、5、10、15、20 mg·L-1)的氧化乐果对小球藻生长发育及EC50,硝酸还原酶活性,叶绿素含量的影响,初步探讨了氧化乐果对小球藻的毒性作用。实验结果表明:氧化乐果对小球藻96 hEC50值为4.88 mg·L-1;低浓度氧化乐果(1 mg·L-1)在前6天对小球藻细胞增长影响不大,高浓度氧化乐果(大于5 mg·L-1)则在初期就表现为抑制,氧化乐果浓度与细胞数量之间表现为负相关(R2=0.65755);在第8天,氧化乐果各浓度组的硝酸还原酶的相对酶活也表现"低促高抑"作用,但是仍然高于对照组的酶活,整体表现为促进作用;随着氧化乐果浓度的增加,小球藻叶绿素含量呈现明显降低的趋势。     

藻类水体Cd2+毒性快速监测新方法研究

采用藻类生长抑制试验和光合活性抑制试验两种方法对蛋白核小球藻受不同浓度Cd2+胁迫作用下叶绿素浓度及藻活性荧光参数进行测量,依据Sigmoidal曲线拟合及单因素方差分析(one-way ANOVA)的方法对Cd2+不同胁迫时间下藻活性抑制率和96 h比生长率抑制率的相关性进行研究.结果表明,48、53、72、77和96 h的藻活性抑制率和96 h比生长率抑制率间具有较好的S函数关系(R2>0.95),因此可采用藻活性抑制率48 h-EC10和53 h-EC10来代替96 h的藻类半数比生长率抑制率EC50进行Cd2+藻类毒性实验评价.进一步分析了蛋白核小球藻受Cd2+胁迫48 h和53 h藻活性抑制率和Cd2+毒性当量的剂量-效应关系.该方法为实验室内单一Cd2+毒性的监测提供了一种快速有效的新方法,为水环境综合毒性预警提供了一种切实可行的方法依据.     

臭氧氧化法深度处理生活垃圾焚烧厂沥滤液

采用臭氧氧化法对生活垃圾焚烧厂沥滤液经生化处理后的废水(称沥滤液生化处理水)进行深度处理。实验结果表明,COD降解速率随废水pH的提高明显增加,其中pH=10.5时的COD降解速率常数约为pH=4时的5.8倍。在臭氧投量为52.92 mg/min、pH=10.5的条件下反应70 min后,UV254和COD去除率分别达到84.7%和59.3%。向反应体系投加叔丁醇后COD去除率下降了约15%,由羟自由基氧化去除的COD占总COD去除量的26.7%。毒性实验结果表明,沥滤液生化处理水的96 h-EC50为38%,经臭氧氧化进一步处理后出水的96 h-EC50为77%,表明经臭氧深度处理后沥滤液生化处理水的毒性明显降低。     

Early stage toxicity of excess copper to photosystem II of Chlorella pyrenoidosa–OJIP chlorophyll a fluorescence analysis

Acute toxicity of excess Cu on the photosynthetic performance of Chlorella pyrenoidosa was examined by using chlorophyll a fluorescence transients and JIP-test after exposure to elevated Cu concentrations for a short time period. High Cu concentration resulted in a significant suppression in photosynthesis and respiration. The absorption flux (ABS/RC) per PSII reaction center increased with increasing Cu concentration, but the electron transport flux (ET₀/RC) decreased. Excess Cu had an insignificant effect on the trapping flux (TR₀/RC). The decline in the efficiency, with which a trapped exciton can move an electron into the electron transport chain further than Q_A−(Ψ₀), the maximal quantum yield of primary photochemistry (?_P0), and the quantum yield of electron transport (?_E0) were also observed. The amount of active PSII reaction centers per excited cross section (RC/CS) was also in consistency with the change of photosynthesis when cells were exposed to excess Cu concentration. JIP-test parameters had a good linear relationship with photosynthetic O₂ evolution. These results suggested that the decrease of photosynthesis in exposure to excess Cu may be a result of the inactivation of PSII reaction centers and the inhibition of electron transport in the acceptor side.     

尼罗红荧光染色法的优化及应用

以自主筛选的小球藻为材料对尼罗红荧光染色进行条件优化,并用该方法追踪微藻在污水中的油脂含量变化。实验发现,微藻只有经过超声波破碎,染色才会出现荧光,DMSO的添加会增强荧光反应。最终确定其染色条件为:DMSO与藻液体积比为1∶5,超声波破碎5 min后染色5 min,NR质量浓度1.5μg/m L。在其应用方面发现,微藻的油脂积累量只有培养基的一半,说明污水不利于微藻油脂的积累;最佳采收时期为平稳期,此时微藻的产量和油脂含量达到最高。     

共生细菌对盐生小球藻亚砷酸盐代谢的影响

为探讨共生细菌对盐生小球藻（Chlorella salina）亚砷酸盐（As（III））富集和形态转化的影响,从C.salina培养液中分离培养出一株共生细菌,并采用抗生素处理C.salina获得无菌藻,设置0,75,150,300,750 μg/LAs（III）溶液处理带菌和无菌的C.salina,7d后测定C.salina对As（III）的吸收、吸附和富集量,以及培养液和藻细胞内As的形态,计算带菌和无菌C.salina对As（III）的氧化率和去除率;使用不同浓度的As（III）处理共生细菌7d,并以300μg/L的As（III）处理共生细菌不同时间,计算两种情况下共生细菌对培养液中As（III）的氧化率和去除率.结果表明：C.salina的共生细菌为根癌农杆菌（Agrobacterium tumefaciens WB-1）;与无菌C.salina相比,带菌C.salina生长更快、对As（III）的耐性更强.带菌C.salina对As（III）的富集量为103.10~448.12mg/kg、氧化率为78.93%~96.88%、去除率为18.92%~55.21%,显著高于无菌C.salina的富集量（11.68~91.39mg/kg）、氧化率（14.46%~26.39%）和去除率（12.82%~29.15%）,也高于A.tumefaciens WB-1对As（III）的氧化率（4.51%~30.61%）和去除率（1.86%~16.19%）.此外,带菌C.salina胞内还检测到少量的As（III）和甲基砷,而在无菌C.salina胞内没有甲基砷存在.共生细菌促进了盐生小球藻对As（III）的富集和形态转化.     

微藻无机氮利用过程中的稳定氮同位素分馏

选用两种δ15N差异显著的铵态氮和硝态氮,分别设置不同浓度的铵态氮和硝态氮来处理莱茵衣藻和蛋白核小球藻,通过分析微藻的稳定氮同位素组成变化,来研究微藻利用不同浓度、不同形态无机氮过程中的稳定氮同位素分馏特征。结果显示,在未添加无机氮的条件下,微藻利用有机氮时,生长缓慢,稳定氮同位素基本上不存在分馏;在添加低浓度无机氮(≤20 mmol/L)时,微藻的生长和稳定氮同位素分馏都随着无机氮浓度的增加而增加;而添加高浓度无机氮(20 mmol/L)时,微藻的生长趋于稳定,铵态氮条件下的微藻稳定氮同位素分馏继续加大,而硝态氮条件下的微藻稳定氮同位素分馏反而减小,可能与此时微藻硝酸还原酶的活力减小有关。