一株高效去除亚硝酸氮细菌的分离鉴定及其脱氮特性研究

【目的】从南美白对虾养殖塘中分离到高效去除亚硝酸氮的细菌, 对其分类和脱氮特性进行了研究。【方法】 以除亚硝酸氮为主要指标, 取养殖塘底层水样分离筛选菌株; 依据16S rRNA基因序列和生理生化特征初步鉴定菌株; 研究不同碳源、碳氮比、起始pH、温度、摇床转速和氯化钠浓度对反硝化除亚硝酸氮的影响, 并考查了菌株对硝酸氮和氨氮的利用情况。【结果】得到的菌株中菌株FP6活性最高, 初步鉴定菌株FP6属于地衣芽孢杆菌。菌株FP6的生长最适脱氮碳源为蔗糖, 菌株FP6去除亚硝酸氮有高活性的条件范围为: C/N值15?25、起始pH 7.0?10.0、温度20 °C?37 °C、摇床转速0?200 r/min和氯化钠浓度0?40 g/L。菌株FP6对硝酸氮和氨氮都有一定的去除能力, 利用硝酸氮时不积累亚硝酸氮。【结论】地衣芽孢杆菌FP6具有优良的除亚硝氮特性, 适宜的温度、pH和盐度范围较宽。     

凡纳滨对虾育苗水体中三种生物絮团的菌群多样性及Tax4Fun基因功能预测分析

为了解凡纳滨对虾(Litopenaeus vannamei)育苗期水体中3种生物絮团的形成过程、水质净化效果、菌群结构及功能, 实验通过测定各组絮团含量和水质指标并采用Illumina MiSeq高通量测序技术, 比较葡萄糖组、淀粉组和蔗糖组生物絮团的不同特征。结果显示, 各组形成的生物絮团可有效调节水质, 降低水体中的氨氮和亚硝酸盐氮水平。在3个碳源添加组水样中氨氮、亚硝酸盐氮和硝酸盐氮浓度显著低于对照组(P<0.05), 在不同碳源组中淀粉组的氨氮、亚硝酸盐氮和硝酸盐氮的浓度显著高于葡萄糖组和蔗糖组(P<0.05); 淀粉组的生物絮团沉降体积(BFV)显著低于葡萄糖组(P<0.05), 蔗糖组BFV最高, 3组粒径大小呈现: 蔗糖组>葡萄糖组>淀粉组; 3组样品分别测得553、515和542个OTU, 菌群丰度指数Chao1、Shannon值葡萄糖组>蔗糖组>淀粉组。各组菌群在门水平中, 以变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和浮霉菌门(Planctomycetes)为各组优势菌门, 3个菌门在各组中占比分别达到91.7%、97.6%和88.7%。在属级水平中, 葡萄糖组中Oceanicella属为优势菌属, 占比最高为18.4%, 淀粉组中丰度较高的Muricauda、Cyclobacterium属, 占比分别为9.8%和5.9%, 均高于葡萄糖组和蔗糖组。在蔗糖组中Rhodopirellula属具有较高占比, 较葡萄糖组和淀粉组分别高1.8%和4.1%。通过Tax4Fun法在3组中检测到细胞新陈代谢、遗传信息加工和环境信息处理等基因功能。这些功能基因在葡萄糖组中的丰度均高于蔗糖组和淀粉组。各项结果表明, 凡纳滨对虾育苗水体中生物絮团不仅可以净化水质, 还能提高水体菌群多样性, 其中以葡萄糖作为碳源效果最好, 生物絮团技术对于维持水质与水体生态系统平衡具有重要意义。     

不同C/N生物絮团对洛氏鱥急性铜暴露保护作用

实验旨在探索不同C/N生物絮团对急性铜暴露洛氏鱥免疫抑制、炎症反应与氧化应激的保护作用。挑选480尾洛氏鱥(Rhynchocypris lagowskii Dybowski)幼鱼(10±0.15) g, 随机分成4组, 每组3个重复, 每个重复40尾鱼。对照组饲喂商品料(C/N 10.8﹕1), 实验组选择葡萄糖为外添碳源调节C/N 15﹕1(Ⅱ组)、C/N 20﹕1(Ⅲ组)和C/N 25﹕1(Ⅳ组), 生长实验为56d, 之后进行为期96h的急性铜暴露胁迫实验。结果表明, 各实验组中免疫、抗氧化酶活性与炎症因子含量随着C/N的增加先升高后下降。其中, 与对照组相比, Ⅲ和Ⅳ组血清碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、溶菌酶(LSZ)、一氧化氮合酶(NOS)、补体C3、C4和免疫球蛋白M(IgM)水平显著升高(P<0.05), 而Ⅲ和Ⅳ组之间补体C3和C4水平差异不显著(P>0.05); Ⅲ和Ⅳ组血清过氧化氢酶(CAT)、抗坏血酸(ASA)、超氧化物歧化酶(SOD)、总抗氧化能力(T-AOC)、谷胱甘肽过氧化物酶(GSH-PX)与谷胱甘肽还原酶(GR)酶活性显著高于对照组(P<0.05), 而丙二醛(MDA)含量显著低于对照组(P<0.05)。同时Ⅱ组的SOD和CAT酶活性也有所升高, 且与对照组差异显著(P<0.05); Ⅲ和Ⅳ组血清肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)与白介素-6(IL-6)含量显著低于对照组(P<0.05), 且它们两组之间无显著差异(P>0.05)。然而, Ⅱ、Ⅲ和Ⅳ组白介素-2(IL-2)含量均显著高于对照组, 且第Ⅲ组含量最高。在实验条件下, 当C/N≥15: 1时, 生物絮团能有效地增强急性铜暴露下洛氏鱥的免疫与抗氧化酶活力, 且在C/N为20﹕1时效果最为显著。     

氨氮对软体动物生长的影响:以铜锈环棱螺为例

为探究氨氮对底栖动物的毒性效应,在位于湖北保安湖的近自然生态系统(单个水域面积约600 m²,水深约1.5 m)中开展了为期1年的模拟实验,分析了6个不同氨氮浓度[N25>N20>N15>N10>N5>N0(对照);0.2—33.7 mg/L]条件下,大型底栖动物(软体动物)群落特征的差异。结果表明:(1)实验系统中采集的软体动物主要为铜锈环棱螺(Bellamya aeruginosa);(2)从B. aeruginosa密度来看, N0、N5、N10和N15处理相差不大[28(0—85) ind./m²],均显著高于N20和N25处理[5(0—29) ind./m²](P<0.05);(3)从B. aeruginosa生物量来看,N0、N5、N10、N15和N20处理相差不大[40.0(0—85.5) g/m²],均显著高于N25处理[0.8(0—4.0) g/m²](P<0.05);(4)从B. aeruginosa壳长、壳宽和带壳...     

接种两株有益菌的生物絮团净水性能及饲料化潜力

为强化新的絮团形成菌伪杜擀氏菌YN12(Pseudoduganella eburnea YN12, YN)利用养殖尾水生成生物絮团效果,并将絮团制成饲料,探讨在胡子鲶(Claris fuscus)饲养上的可行性。在生物絮团系统中,接种活性污泥(Activated sludge, AS)、伪杜擀氏菌YN12和枯草芽孢杆菌(Bacillus subtilis, BS)并配制成7组(AS、YN、BS、AS+YN、AS+BS、YN+BS和AS+YN+BS),分析絮团形态结构差异,评估出对模拟养殖尾水氮去除效果及微生物群落结构优势的最佳组合,最终收集最佳组合絮团烘干研磨成粉末添加到商业饲料制粒,分成对照组和絮团组两组,探讨对胡子鲶的生长性能、饲料利用率、肌肉营养成分及肝脏的消化酶和免疫酶活性的影响。结果表明:YN+BS组水质净化效果稳定,总氮去除效果(89.6%)良好,氨氮、亚硝酸盐和硝酸盐积累低,具有良好的絮团生成量(9.0 ml/L)及占据优势(47.5%)的反硝化菌群(包括Hydrogenophaga、Flavobacterium、Pseudoxanthomonas、Burkholderi...     

土壤碳氮比对平邑甜茶幼苗生长和碳氮分配的影响

为探讨土壤碳氮比(C:N)对苹果(Malus pumila)植株生长和碳氮分配特性的影响, 采用碳氮双标记示踪技术, 以二年生平邑甜茶(Malus hupehensis)幼苗为试验材料, 研究了6个不同土壤C:N处理(T1-T6分别为4.70、9.78、14.70、19.96、25.60和28.83)下平邑甜茶的生长状况和氮素吸收、利用分配以及碳水化合物的运转特性。结果表明, 随着土壤C:N的逐渐增大, 平邑甜茶幼苗根系干重逐渐增加, 而株高、茎粗、地上部干重和植株总干重呈先增加后降低的趋势, 以T4处理最大。土壤C:N显著影响了平邑甜茶幼苗的 ¹⁵N利用率, 从T1到T4处理, 植株的 ¹⁵N利用率逐渐升高, T4处理(18.46%)是T1处理(10.65%)的1.73倍; 随着土壤C:N的进一步增加, 植株的 ¹⁵N利用率逐渐降低, T5和T6处理分别比T4处理降低了1.59%和2.58%。土壤C:N较低的T1和T2处理, 平邑甜茶幼苗各器官从肥料中吸收分配到的 ¹⁵N量对该器官全氮量的贡献率(Ndff)大小顺序为根>叶>茎, 随着土壤C:N的进一步增大, 叶片的Ndff均为最大, 其次是根, 茎最少。随着土壤C:N的增大, 叶片 ¹⁵N分配率逐渐升高, ¹³C分配率逐渐降低; 而根系 ¹⁵N分配率逐渐降低, ¹³C分配率逐渐升高。综合考虑植株生长和氮素利用状况, 本试验条件下适宜平邑甜茶生长的土壤C:N为21-23。     

温度、pH、溶氧对添加植物乳杆菌构建生物絮凝系统的影响

生物絮凝技术和微生物制剂在水产养殖业中广泛应用,但向生物絮凝系统中添加益生菌最优条件研究甚少。本试验采取L_9(3~3)安排3因素3水平关于溶氧(厌氧0.5 mg/L,有氧5 mg/L,厌氧-有氧交替)、pH (5.5, 7, 8.5)、温度(25℃, 30℃, 35℃)正交试验,开展35 d以植物乳杆菌为添加剂培养生物絮团试验,通过比较三态氮的变化情况、脱氢酶活性(dehydrogenase activity, DHA)及微生物种群结构,得出以植物乳杆菌为添加剂培养生物絮凝的最佳工艺。结果表明:在有氧、pH值8.5、30℃时,生物絮凝培养用时最短,为13 d,生物脱氮效率高;温度和溶氧对脱氢酶活性有显著性影响,25℃,厌氧-好氧交替,pH值7条件下培养的生物絮团脱氢酶活性最高;微生物群落结构分析表明生物絮团中主要为变形菌门(70%),其次为拟杆菌门;植物乳杆菌在有氧、35℃、pH值5.5条件下丰度最高,为1.8%。综合实际生产,添加植物乳杆菌构建生物絮凝系统最佳条件为30℃、p H值7、有氧条件(DO5 mg/L)。     

生物土壤结皮演替对土壤生态化学计量特征的影响

生物土壤结皮(生物结皮)是干旱半干旱地区重要的地表覆被物,能够固定碳氮,影响养分循环,从而可能引起土壤化学计量特征的变化。以黄土丘陵区不同演替阶段生物结皮为研究对象,研究该区生态恢复初期生物结皮演替对土壤碳(C)、氮(N)、磷(P)含量及其化学计量特征的影响。结果表明:(1)生物结皮显著增加了生物结皮层C、N、P含量,后期藓结皮较初期藻结皮C、N、P含量分别增加了161%、127%和9%,均显著高于其下0—10 cm土壤;(2)土壤C/N随着生物结皮演替变化较小,范围为10.0—11.8,C/P和N/P在演替后期分别是初期的2.4倍和2.1倍,均显著高于其下0—10 cm土壤;(3)生物结皮土壤C/N、C/P与N/P受坡向影响较大,并与藓生物量显著正相关,与土壤容重显著负相关;(4)生物结皮显著影响土壤C、N、P含量及其化学计量特征,使生物结皮层显著不同于其下层土壤及裸地;(5)生物结皮演替加速表层土壤养分恢复,影响程度可至其下2 cm土层。研究初次从土壤化学计量特征的角度揭示生物结皮对土壤养分循环的贡献,对干旱半干旱地区生态修复和管理具有指导意义。     

碳氮比对模拟水稻秸秆田间厌氧发酵系统甲烷生产和秸秆降解的影响

利用自制的厌氧发酵装置模拟田间厌氧发酵产生物甲烷的条件,通过添加不同量的尿素,研究C/N对水稻秸秆厌氧发酵产生物甲烷和秸秆降解的影响。设置了C/N为15∶1、20∶1、25∶1、30∶1和对照(53∶1)等五个处理,测定各处理的产气量、甲烷含量和秸秆木质纤维素含量。结果表明C/N对厌氧发酵产气进程有一定影响,峰值出现的时间、峰值的大小在不同处理之间均有差异:以C/N为25∶1的处理峰值出现的最早,较对照早5 d出现;以C/N为20∶1的峰值最高,较对照提高了27. 55%。C/N对厌氧发酵累积产气量和累积甲烷产量有显著影响,以C/N为20∶1的处理累积产气量(4 235. 00 mL)和累积甲烷产量(1526. 34mL)最高,相比对照组的累积产气量和累积甲烷产量分别提高了30. 78%和51. 31%。C/N对水稻秸秆降解也有显著影响。以20∶1和25∶1处理对秸秆降解的效果最好,其中20∶1处理的总降解率、纤维素降解率和半纤维素降解率分别达到了51. 33%、55. 31%和53. 05%,25∶1处理分别为50. 00%、53. 08%和49. 42%。上述结果表明通过添加尿素调节C/N能够提高水稻秸秆厌氧发酵的产生物甲烷效率和促进秸秆降解,以C/N为(20～25)∶1处理的促进效果最好。     

异养硝化-好氧反硝化菌Acinetobacter johnsonii sp. N26的脱氮性能及代谢途径

【背景】水体中含氮物质的大量累积会造成水体富营养化、水生生物死亡等问题,严重威胁水生态环境,制约我国环境保护的持续发展。【目的】为去除生活污水中的含氮污染物,从羊粪堆肥中筛选出了一株具有异养硝化-好氧反硝化功能的细菌——约氏不动杆菌Acinetobacter johnsonii sp.N26,研究其脱氮性能和代谢途径。【方法】测定菌株N26在氨氮和硝态氮中的生长和脱氮曲线,通过单因素试验对其脱氮性能进行优化,通过氮平衡分析和功能基因鉴定研究其脱氮代谢途径。【结果】生长和脱氮曲线表明,菌株N26对初始浓度均为50 mg/L的氨氮和硝态氮的去除速度快、效率高,其中9 h内对氨氮的去除效率为95.5%,最大去除速率为5.330 mg/(L·h);15 h内对硝态氮的去除效率为93.6%,最大去除速率为3.147 mg/(L·h),且最终仅有少量硝酸盐、亚硝酸盐积累。脱氮性能优化结果表明,该菌株的最适氮源为氯化铵,最适碳源为丁二酸钠,最适温度为30℃,最适接种量为15%,最适p H值为8.0-9.0,最适碳氮比为15,最适转速为120 r/min,最适氮负荷≤300 mg/L (氨氮)。氮平衡...     

Effects of Temperature and Carbon-Nitrogen (C/N) Ratio on the Performance of Anaerobic Co-Digestion of Dairy Manure,Chicken Manure and Rice Straw: Focusing on Ammonia Inhibition

Anaerobic digestion is a promising alternative to disposal organic waste and co-digestion of mixed organic wastes has recently attracted more interest. This study investigated the effects of temperature and carbon-nitrogen (C/N) ratio on the performance of anaerobic co-digestion of dairy manure (DM), chicken manure (CM) and rice straw (RS). We found that increased temperature improved the methane potential, but the rate was reduced from mesophilic (30∼40°C) to thermophilic conditions (50∼60°C), due to the accumulation of ammonium nitrogen and free ammonia and the occurrence of ammonia inhibition. Significant ammonia inhibition was observed with a C/N ratio of 15 at 35°C and at a C/N ratio of 20 at 55°C. The increase of C/N ratios reduced the negative effects of ammonia and maximum methane potentials were achieved with C/N ratios of 25 and 30 at 35°C and 55°C, respectively. When temperature increased, an increase was required in the feed C/N ratio, in order to reduce the risk of ammonia inhibition. Our results revealed an interactive effect between temperature and C/N on digestion performance.     

马尾松林两种林下植被土壤碳氮特征及其与凋落物质量的关系

以飞播马尾松林为研究对象,通过典型样地调查和样品测定,采用配对样本t检验和冗余分析（RDA）方法分析芒萁类和禾草类两种林下植被类型土壤碳、氮特征及其与凋落物质量之间的关系。结果表明：（1）土壤有机碳、微生物量碳、可溶性有机碳、全氮、速效氮、微生物量氮和可溶性有机氮含量在0-10、10-20 cm土层均表现为禾草类显著高于芒萁类（P < 0.05）,而在20-40、40-80 cm土层两种植被类型碳氮指标的大小未表现出相同的变化规律,且差异不显著（P > 0.05）。（2）两种植被类型凋落物半分解和未分解层的C含量及C/N值均表现为芒萁类显著高于禾草类（P < 0.05）,而N含量则表现为禾草类显著高于芒萁类（P < 0.05）;同一植被类型的未分解层C含量及C/N值均显著大于半分解层,N含量则半分解层显著大于未分解层（P < 0.05）。（3）0-10 cm土层两种类型凋落物C/N值和C含量均与土壤碳氮各指标呈显著负相关（P < 0.05）,N含量与土壤碳氮各指标的相关性不显著（P > 0.05）;10-20 cm土层,芒萁类的半分解层C/N值与土壤碳氮各指标存在显著相关性（P < 0.05）,禾草类的凋落物C含量与土壤碳氮各指标也存在显著相关性（P < 0.01）。林下植被凋落物C/N值越小,其分解速率越快,有利于土壤养分的积累,禾草类凋落物C/N值低于芒萁类是导致其土壤碳氮指标高于芒萁类的重要原因。     

Environmental remediation potential of Perinereis aibuhitensis (Polychaeta) based on the effects of temperature and feed types on its carbon and nitrogen budgets

The effects of three temperatures (15°C, 20°C and 25°C) and four feed types (artificial feed, AF; sediment, SM; fish faeces, FF; and fish meat, FM) on the growth, carbon budget and nitrogen budget of the polychaete Perinereis aibuhitensis were evaluated over a 35 day period. The final body weight and specific growth rate (SGR) of P. aibuhitensis were significantly affected by temperature and feed types (T, F?=?15.831, P?=?0.000; Feed, F?=?81.827, P?=?0.000), but the interaction between these factors was not significant (F?=?0.435, P?=?0.848). The worms achieved the highest SGR in the AF group at 20°C. However, the SGR in the SM group was only half that of the other groups. The food conversion efficiency (FCE) was significantly lower at 25°C than at 15°C and 20°C (P?P?=?0.000), with a mean of 39.83%. The apparent digestibility rate (ADR) at 25°C was significantly higher than at 15°C and 20°C (P?相似文献   

Effect of sublethal concentrations of four chemicals on susceptibility of juvenile rainbow trout (Oncorhynchus mykiss) to saprolegniosis.

The effects of sublethal concentrations of a variety of chemicals on the susceptibility of rainbow trout (Oncorhynchus mykiss) juveniles to Saprolegnia parasitica infection was examined. Sublethal concentrations of un-ionized ammonia (0.05 mg/liter) and nitrite (0.12 mg/liter) increased fish susceptibility after 10 days of exposure to the toxin, this increase being higher for ammonia (75% and 20% morbidity) than for nitrite (20% and 0% morbidity, respectively) with inoculum doses of 1.4 x 10(6) and 9.5 x 10(5) zoospores per liter, respectively. Sublethal concentrations of copper (0.05 mg/liter) or cyanide (0.05 mg/liter) did not show enhancement of infection by S. parasitica, even though the toxin exposure was for 21 days and the inoculum doses were higher than those for the experiments with the nitrogen compounds (4 x 10(6) and 3.2 x 10(6) zoospores per liter. However, infections began to appear in control animals.     

Effect of sublethal concentrations of four chemicals on susceptibility of juvenile rainbow trout (Oncorhynchus mykiss) to saprolegniosis

The effects of sublethal concentrations of a variety of chemicals on the susceptibility of rainbow trout (Oncorhynchus mykiss) juveniles to Saprolegnia parasitica infection was examined. Sublethal concentrations of un-ionized ammonia (0.05 mg/liter) and nitrite (0.12 mg/liter) increased fish susceptibility after 10 days of exposure to the toxin, this increase being higher for ammonia (75% and 20% morbidity) than for nitrite (20% and 0% morbidity, respectively) with inoculum doses of 1.4 x 10(6) and 9.5 x 10(5) zoospores per liter, respectively. Sublethal concentrations of copper (0.05 mg/liter) or cyanide (0.05 mg/liter) did not show enhancement of infection by S. parasitica, even though the toxin exposure was for 21 days and the inoculum doses were higher than those for the experiments with the nitrogen compounds (4 x 10(6) and 3.2 x 10(6) zoospores per liter. However, infections began to appear in control animals.     

Growth of Nitrosomonas europaea on hydroxylamine

Abstract Hydroxylamine is an intermediate in the oxidation of ammonia to nitrite, but until now it has not been possible to grow Nitrosomonas europaea on hydroxylamine. This study demonstrates that cells of N. europaea are capable of growing mixotrophically on ammonia and hydroxylamine. The molar growth yield on hydroxylamine (4.74 g mol⁻¹ at a growth rate of 0.03 h⁻¹) was higher than expected. Aerobically growing cells of N. europaea oxidized ammonia to nitrite with little loss of inorganic nitrogen, while significant inorganic nitrogen losses occurred when cells were growing mixotrophically on ammonia and hydroxylamine. In the absence of oxygen, hydroxylamine was oxidized with nitrite as electron acceptor, while nitrous oxide was produced. Anaerobic growth of N. europaea on ammonium, hydroxylamine and nitrite could not be observed at growth rates of 0.03 h⁻¹ and 0.01 h⁻¹.     

Effect of carbon to nitrogen (C:N) ratio on nitrogen removal from shrimp production waste water using sequencing batch reactor

The United States Marine Shrimp Farming Program (USMSFP) introduced a new technology for shrimp farming called recirculating raceway system. This is a zero-water exchange system capable of producing high-density shrimp yields. However, this system produces wastewater characterized by high levels of ammonia, nitrite, and nitrate due to 40% protein diet for the shrimp at a high density of 1,000 shrimp per square meter. The high concentrations of nitrate and nitrite (greater than 25 ppm) are toxic to shrimp and cause high mortality. So treatment of this wastewater is imperative in order to make shrimp farming viable. One simple method of treating high-nitrogen wastewater is the use of a sequencing batch reactor (SBR). An SBR is a variation of the activated sludge process, which accomplishes many treatment events in a single reactor. Removal of ammonia and nitrate involved nitrification and denitrification reactions by operating the SBR aerobically and anaerobically in sequence. Initial SBR operation successfully removed ammonia, but nitrate concentrations were too high because of carbon limitation in the shrimp production wastewater. An optimization study revealed the optimum carbon to nitrogen (C:N) ratio of 10:1 for successful removal of all nitrogen species from the wastewater. The SBR operated with a C:N ratio of 10:1 with the addition of molasses as carbon source successfully removed 99% of ammonia, nitrate, and nitrite from the shrimp aquaculture wastewater within 9 days of operation.