生物电化学耦合厌氧氨氧化强化脱氮及其微生物群落特征

为探究生物电化学强化厌氧氨氧化(anaerobic ammonia oxidation,anammox)脱氮作用过程,采用双室微生物电解池(microbial electrolysis cell,MEC)富集电活性微生物,构建耦合厌氧氨氧化阴极的生物电化学系统。具体地,在外加0.2 V电压条件下改变不同总氮进水浓度于30°C进行暗培养批次实验研究,结合循环伏安法、电化学阻抗谱、高通量测序方法等多种表征手段研究了强化脱氮机理。结果表明,在初始总氮浓度分别为200、300和400 mg/L时对应获得了96.9%±0.3%、97.3%±0.4%和99.0%±0.3%的总氮去除率,且阴极电极生物膜表现出良好的电化学活性。高通量测序结果表明外加电压富集了除厌氧氨氧化菌以外的其他脱氮功能菌群:反硝化菌(Denitratisoma)、Limnobacter和氨氧化菌SM1A02和Anaerolineaceae、亚硝化菌(Nitrosomonas europaea)和硝化螺菌属(Nitrospira)等,这些具有电化学活性的微生物构成了体系的氨氧化胞外产电菌(ammonium oxidizing exoelectrogens,AOE)和反硝化电养菌(denitrifying electrotrophs,DNE),它们连同厌氧氨氧化菌Candidatus Brocadia构成了系统的脱氮微生物群落结构。AOE和DNE的种间直接电子传递作用协同厌氧氨氧化是强化系统总氮去除的关键原因。     

有机碳源下废水厌氧氨氧化同步脱氮除碳

为明确有机碳源胁迫下,厌氧氨氧化反应器的同步脱氮除碳规律及功能微生物群落结构的动态变化,采用成功启动的厌氧氨氧化UASB反应器,通过逐步提升进水有机负荷,探究有机碳源下废水厌氧氨氧化同步脱氮除碳。研究表明,当进水化学需氧量(Chemical oxygen demand,COD)浓度从172 mg/L升至620 mg/L,反应器维持较高的脱氮效率,氨氮和总氮去除率均在85%以上,并对COD具有平均56.6%的去除率,高浓度COD未对Anammox菌活性构成显著抑制作用。聚合酶链式反应和变性梯度凝胶电泳(PCR-DGGE)图谱和割胶测序结果表明,变形菌门Proteobacteria、浮霉菌门Planctomycetes、绿曲挠菌门Chloroflexi以及绿菌门Chlorobi等微生物共存于同一反应体系中,推测反应器内存在复杂的脱氮除碳途径。而且,代表厌氧氨氧化的部分浮霉菌门微生物能耐受高浓度有机碳源,在高有机负荷下依旧发挥着高效的脱氮作用,为反应器高效脱氮提供了保障。     

Stimulatory effects of low intensity ultrasound on the growth kinetics and metabolic activity of Lactococcus lactis subsp. Lactis

The present study was carried out to investigate the effects of low intensity ultrasound (20% and 30% amplitudes for 3 min and 5 min) on the growth kinetics and metabolic activity of Lactococcus lactis subsp. Lactis. The results revealed that the viable cell counts increased by 6.21–21.82% under ultrasound treatment. Confocal scanning laser microscopy showed 3.03–22.88% reduction in the percentage of intact cells after ultra-sonication. The specific growth rate of ultrasonicated L. lactis subsp. Lactis also increased by 7.09–23.22%. The pH values and titratable acidity were lower and higher, respectively, for most treatments. The treatments increased β-galactosidase activity. This was accompanied with a faster rate in lactic acid yield. The protein concentrations significantly decreased with increasing ultrasound amplitude and duration of irradiation, whilst at the same conditions the degree of protein hydrolysis was higher. Results suggest that ultrasound can stimulate the growth of L. lactis subsp. Lactis and its metabolic activity. Therefore, ultrasound can potentially play a significant role in stimulating fermentative activities of starter cultures and may successfully be employed in fermentation industries.     

Cold shocks of Anammox biofilm stimulate nitrogen removal at low temperatures

The adaptation of Anammox (ANaerobic AMMonium OXidation) to low temperatures (10–15°C) is crucial for sustaining energy‐efficient nitrogen removal from the mainstream of municipal wastewater. But, current adaptation methods take months or even years. To speed up the adaption of Anammox to low temperatures, this study describes a new approach: exposing Anammox microorganisms to an abrupt temporary reduction of temperature, i.e., cold shock. Anammox biomass in a moving bed biofilm reactor was subjected to three consecutive cold shocks (reduction from 24 ± 2 to 5.0 ± 0.2°C), each taking eight hours. Before the cold shocks, Anammox activity determined in ex situ tests using the temperature range of 12.5–19.5°C was 0.005–0.015 kg‐N kg‐VSS^?1 day^?1. Cold shocks increased the activity of Anammox at 10°C to 0.054 kg‐N kg‐VSS^?1 day^?1 after the third shock, which is similar to the highest activities obtained for cold‐enriched or adapted Anammox reported in the literature (0.080 kg‐N kg‐VSS^?1 day^?1). Fluorescence in situ hybridization analysis showed that Ca. Brocadia fulgida was the dominant species. Thus, cold shocks are an intriguing new strategy for the adaptation of Anammox to low temperature. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:277–281, 2018     

New concepts of microbial treatment processes for the nitrogen removal in wastewater

Many countries strive to reduce the emissions of nitrogen compounds (ammonia, nitrate, NOx) to the surface waters and the atmosphere. Since mainstream domestic wastewater treatment systems are usually already overloaded with ammonia, a dedicated nitrogen removal from concentrated secondary or industrial wastewaters is often more cost-effective than the disposal of such wastes to domestic wastewater treatment. The cost-effectiveness of separate treatment has increased dramatically in the past few years, since several processes for the biological removal of ammonia from concentrated waste streams have become available. Here, we review those processes that make use of new concepts in microbiology: partial nitrification, nitrifier denitrification and anaerobic ammonia oxidation (the anammox process). These processes target the removal of ammonia from gases, and ammonium-bicarbonate from concentrated wastewaters (i.e. sludge liquor and landfill leachate). The review addresses the microbiology, its consequences for their application, the current status regarding application, and the future developments.     

Establishment of axon regeneration regulatory network and the role of low intensity pulsed ultrasound in the network

ObjectiveTo establish an axon regeneration regulatory network for optimal selection, and explore the role of low intensity pulsed ultrasound in the network.MethodsThe axon regeneration regulatory network involving axon regeneration-related proteins NGF, BDNF and PirB was constructed by using GO and KEGG. The maximum possible pathway acting on axon regeneration was screened by Bayesian network theory. The node of low - intensity pulsed ultrasound in NGF - involved axon regeneration network was complemented by combining literature methods.ResultsThe NGF, BDNF and PirB-involved axonal regeneration regulatory pathway was successfully constructed. The low intensity pulsed ultrasound played a role in axon regeneration by acting on ERK1/2-CREB pathway and GSK-3β. NGF-TrKA-Rap1-ERK1/2-CREB-Bcl-2 was optimized as optimal pathway by Bayesian theory.ConclusionThe regulatory pathway of axon regeneration involving nerve growth related factors and low intensity pulsed ultrasound was initially established, which provided a theoretical basis for further study of axon regeneration, and also new ideas for action of low intensity pulsed ultrasound on axon regeneration regulatory pathway.     

低强度脉冲超声波辐照与泡沫TiC/Ti对犬节段性骨缺修复的促进作用

目的探讨低强度脉冲超声波辐照对节段性骨缺损修复效果的影响。方法将直径12 mm长20mm泡沫TiC/Ti植入6只Beagle犬的左侧胫骨节段性骨缺损区。随机分为超声组和对照组,超声组采用低强度脉冲超声波辐照（频率1.5 MHz、强度30 mW/cm2、脉冲宽度200μs、脉冲周期1 kHz、20 min/次、1次/d）,对照组为不开功率源的假辐照,术后4、8周后分别行X线检查及骨密度测定,观察及分析材料周围骨愈合情况。结果 6只beagle犬均进入结果分析。术后4周超声组骨早期成熟度优于对照组,表现在材料周围骨痂影密度增高,骨痂影由两端向中央生长;对照组仅见骨痂区密度低,还可见部分骨痂缺如。术后8周超声组新生骨痂面积优于对照组,骨干结构相对稳定;对照组骨缺损区未闭合,在骨干两侧看到少量骨痂,愈合较差。骨密度测定结果显示,4周时超声组高于对照组,两组间存在统计学差异;8周时超声组略高于对照组,但两组间没有统计学差异。结论通过联合应用低强度脉冲超声波辐照与人工骨材料修复可提高新骨形成速度及骨组织密度,缩短节段性骨缺损的骨愈合时间。     

Analysis of microbial communities developed on the fouling layers of a membrane-coupled anaerobic bioreactor applied to wastewater treatment

The structure of the biofouling layers formed on a pilot-scale membrane-coupled upflow anaerobic sludge blanket bioreactor (UASB) used to treat urban wastewater was analyzed by scanning electron microscopy and electron-dispersive X-ray microanalysis. For comparison, control samples of the membranes were fed either UASB effluent or raw wastewater in a laboratory-scale experiment. Microbial diversity in the fouling materials was analyzed by temperature gradient gel electrophoresis (TGGE) combined with sequence analysis of partial 16S rRNA. Significant differences in structure of the Bacteria communities were observed amongst the different fouling layers analyzed in the UASB membranes, particularly following a chemical cleaning step (NaClO), while the Archaea communities retained more similarity in all samples. The main Bacteria populations identified were evolutively close to Firmicutes (42.3%) and Alphaproteobacteria (30.8%), while Archaea were mostly affiliated to the Methanosarcinales and Methanospirillaceae. Sphingomonadaceae-related bacteria and methanogenic Archaea were persistently found as components of biofouling, regardless of chemical cleaning.     

A study of the spatial organisation of microbial cells in a gel matrix subjected to treatment with ultrasound standing waves

Retention and manipulation of microbial cells through exploitation of ultrasonic forces has been reported as a novel cell immobilisation technique. The spatial ordering of yeast cells, within suspensions subjected to an ultrasonic standing wave field, was analysed for the first time. A technique, based on `freezing' the spatial arrangement using polymer gelation was developed. The resultant gel was then sectioned and examined using microscopic techniques. Light Microscopy confirmed the presence of specific regions in the ultrasonic field, where the cells are organised into bands corresponding to the standing waves' pressure nodal planes. Computer Image Analysis measurement of several physical parameters associated with this cell distribution matched the values derived from the theoretical model. The spatial cell-cell re-arrangement within each band and uneven distribution along the nodal planes have been analysed by Scanning Electron Microscopy. These results complement the ongoing study of the process of immobilisation of microbial cells by ultrasound standing waves. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of the antifungal activity of Rumex vesicarius L. and Ziziphus spina-christi (L) Desf. Aqueous extracts and assessment of the morphological changes induced to certain myco-phytopathogens

Many Plant extracts had proved a potential antifungal activity against a wide range of phytopathogenic fungi. The aim of this study was to evaluate the antifungal activity of the aqueous extracts of Rumex vesicarius L. and Ziziphus spina-christi (L) Desf. against some fungal species. The effect on growth inhibition, conidia germination, sporogenesis, morphological, and ultrastructural characterizations of fungal growth by scanning and transmission electron microscopes, have been investigated. Both plant extracts exhibited an antifungal activity against Fusarium, Helminthosporium, Alternaria, and Rhizoctonia species, besides, the sporogenesis of Alternaria and Fusarium species was suppressed. Both plants induced severe morphological changes in the hyphal shape and surface. We concluded that the aqueous extracts of these plants had strong antifungal activities. More investigations should be performed to evaluate the possible applications in agriculture and in vivo.     

Integration of a Coagulation/Flocculation step in a biological sequencing batch reactor for COD and nitrogen removal of supernatant of anaerobically digested piggery wastewater

The supernatant from mesophilic anaerobic digestion of piggery wastewater is characterised by a high amount of COD (4.1 g COD L(-1)), ammonium (2.3g NH(4)(+)-NL(-1)) and suspended solids (2.5 g SS L(-1)). This effluent can be efficiently treated by means of a Sequencing Batch Reactor (SBR) strategy for biological COD, SS and nitrogen removal including a Coagulation/Flocculation step. Total COD and SS reduction yields higher than 66% and 74%, respectively, and a total nitrogen removal (via nitrite) of more than 98% were reached when working with HRT 2.7 days, SRT 12 days, temperature 32 degrees C, three aerobic/anoxic periods, without external control of pH and under limited aeration flow. The inhibition of nitrite oxidizing biomass was achieved by the working free ammonia concentration and the restricted air supply (dissolved oxygen concentration below 1 mg O(2)L(-1)). Since a part of the total COD was colloidal and/or refractory, a Coagulation/Flocculation step was implemented inside the SBR operating strategy to meet a suitable effluent quality to be discharged. Several Jar-Tests demonstrated that the optimal concentration of FeCl(3) was 800 mg L(-1). A respirometric assay showed that this coagulant dosage did not affect the biological activity of nitrifying/denitrifying biomass.     

生物膜型污水脱氮系统中膜结构及微生物生态研究进展

生物膜法污水脱氮系统主要利用生物膜中脱氮功能微生物的代谢活动去除氮素,从而达到净化水质的目的,研究脱氮生物膜的微观结构和微生物生态是揭示生物膜脱氮机理从而提高脱氮效率的重要途径.本文综述了生物膜型污水脱氮系统类型、生物膜微观结构特征及其影响因素、生物膜型污水脱氮系统内氮素传质过程、脱氮机理和生物膜数学模型等方面的研究进展.另外,本文介绍了生物膜型污水脱氮系统内生物膜脱氮功能微生物分布特征,不同生物膜脱氮系统、底物、运行条件和时间对功能微生物群落影响,及新型脱氮功能微生物等方面的研究进展,为生物膜脱氮技术的深入研究提供参考.     

Plackett-Burman Design与均匀设计法优化玫瑰色微球菌固定化脱氮的性能

【目的】采用Plackett-Burman Design (PBD)与均匀设计(Uniform Design, UD) 优化玫瑰色微球菌(Micrococcus roseus)固定化脱氮的条件,以提高其脱氮性能。【方法】选择聚乙烯醇(Polyvinyl alcohol,PVA)与海藻酸钠(Sodium alginate,SA)作为固定化细胞材料,研究其成球方式与性能,应用PBD对9个影响脱氮效果的因素进行显著性检验,筛选出PVA、SA、温度、菌液接种量、菌球量5个显著性因素(P<0.05)。采用UD对5个显著因素进行条件优化,优化数据通过Minitab与Matlab软件进行逐步回归分析,并建立二阶多项式模型。【结果】最优脱氮条件为PVA 9.6%、SA 2.2%、温度33.4 °C、菌液接种量8%、菌球量500个/100 mL,连续脱氮72 h,最高脱氮率达到60.9%。【结论】PBD与UD的联合使用能够在显著因子筛选与寻优中发挥重要的作用。连续脱氮试验表明,玫瑰色微球菌去除氨氮主要通过同化、硝化与反硝化途径。     

青藏高原东缘窄叶鲜卑花灌丛生长季土壤无机氮对增温和植物去除的响应

为了揭示气候变暖背景下高寒灌丛土壤氮转化过程, 该文研究了青藏高原东缘窄叶鲜卑花(Sibiraea angustata)灌丛生长季节土壤硝态氮和铵态氮含量对增温和去除植物的响应。结果表明: 窄叶鲜卑花灌丛土壤硝态氮和铵态氮含量具有明显的季节动态。整个生长季节, 土壤硝态氮含量呈先增加后降低的趋势, 而铵态氮含量均表现为一直增加的趋势。在生长季初期和中期, 各处理土壤硝态氮含量均显著高于铵态氮含量, 而在生长季末期土壤硝态氮含量均显著低于铵态氮含量, 说明该区域土壤氮转化过程在生长季初期和中期以硝化作用为主, 而在生长季末期以氨化作用为主。不同时期土壤硝态氮和铵态氮含量对增温和去除植物的响应不同: 增温对硝态氮的影响主要发生在生长季中期和末期, 且因植物处理的不同而有显著差异, 增温仅在生长季中期使不去除植物样方铵态氮含量显著升高。去除植物对土壤硝态氮的影响仅表现在对照样方(不增温), 去除植物显著提高了生长季初期和中期土壤硝态氮含量, 显著降低了生长季末期土壤硝态氮含量; 同时去除植物显著降低了增温样方生长季中期土壤铵态氮含量。灌丛植被在生长季初期和中期可能主要吸收土壤硝态氮, 其吸收过程不受土壤增温的影响。     

Impact of low‐intensity pulsed ultrasound on the growth of Schizochytrium sp. for omega‐3 production

Schizochytrium sp. is a microalga that is known for its high content of oils or lipids. It has a high percentage of polyunsaturated fatty acids in the accumulated oil, especially docosahexaenoic acid (DHA). DHA is an important additive for the human diet. Large‐scale production of Schizochytrium sp. can serve as an alternative source of DHA for humans as well as for fish feed, decreasing the burden on aqua systems. Therefore, research on improving the productivity of Schizochytrium attracts a lot of attention. We studied the potential of using low‐intensity pulsed ultrasound (LIPUS) in the growth cycle of Schizochytrium sp. in shake flasks. Different intensities and treatment durations were tested. A positive effect of LIPUS on biomass accumulation was observed in the Schizochytrium sp. culture. Specifically, LIPUS stimulation at the ultrasound intensity of 400 mW/cm² with 20 min per treatment 10 times a day with equal intervals of 2.4 h between the treatments was found to enhance the growth of Schizochytrium biomass most effectively (by up to 20%). Due to the nature of cell division in Schizochytrium sp. which occurs via zoospore formation, LIPUS stimulation was inefficient if applied continuously during all 5 days of the growth cycle. Using microscopy, we studied the interval between zoospore formation in the culture and selected the optimal LIPUS application days (Days 0–1 and Days 4–5 of the 5‐day growth cycle). Microscopic images have also shown that LIPUS stimulation enhances zoospore formation in Schizochytrium sp., leading to more active cell division in the culture. This study shows that LIPUS can serve as an additional tool for cost‐efficiency improvement in the large‐scale production of Schizochytrium as a sustainable and environmentally friendly source of omega‐3 (DHA).     

Redox-stratification controlled biofilm (ReSCoBi) for completely autotrophic nitrogen removal: the effect of co- versus counter-diffusion on reactor performance

A multi-population biofilm model for completely autotrophic nitrogen removal was developed and implemented in the simulation program AQUASIM to corroborate the concept of a redox-stratification controlled biofilm (ReSCoBi). The model considers both counter- and co-diffusion biofilm geometries. In the counter-diffusion biofilm, oxygen is supplied through a gas-permeable membrane that supports the biofilm while ammonia (NH(4)(+)) is supplied from the bulk liquid. On the contrary, in the co-diffusion biofilm, both oxygen and NH(4)(+) are supplied from the bulk liquid. Results of the model revealed a clear stratification of microbial activities in both of the biofilms, the resulting chemical profiles, and the obvious effect of the relative surface loadings of oxygen and NH(4)(+) (J(O(2))/J(NH(4)(+))) on the reactor performances. Steady-state biofilm thickness had a significant but different effect on T-N removal for co- and counter-diffusion biofilms: the removal efficiency in the counter-diffusion biofilm geometry was superior to that in the co-diffusion counterpart, within the range of 450-1,400 microm; however, the efficiency deteriorated with a further increase in biofilm thickness, probably because of diffusion limitation of NH(4)(+). Under conditions of oxygen excess (J(O(2))/J(NH(4)(+)) > 3.98), almost all NH(4)(+) was consumed by aerobic ammonia oxidation in the co-diffusion biofilm, leading to poor performance, while in the counter-diffusion biofilm, T-N removal efficiency was maintained because of the physical location of anaerobic ammonium oxidizers near the bulk liquid. These results clearly reveal that counter-diffusion biofilms have a wider application range for autotrophic T-N removal than co-diffusion biofilms.     

苗龄、光照强度和施氮量对抗褐飞虱水稻品种主要防御酶活性的影响

为探明苗龄、光照强度和施氮量对水稻主要防御酶活性的影响及其与抗褐飞虱的关系,分别就苗龄、光照强度和施氮量等因子对不同抗褐飞虱水稻品种苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)和过氧化物酶(POD)活性的影响进行研究。结果表明:IR56、Ptb33和570011对褐飞虱生物型II的抗性与PAL和PPO活性密切相关,RHT对II型的抗性与PAL、PPO和POD活性密切相关,RP1976-18-6-4-2对II型的抗性与PAL活性密切相关,570011对II型的抗性与PAL和PPO活性密切相关;IR56对褐飞虱孟加拉型的抗性与PAL、PPO和POD活性密切相关,Rathu Heenati(RHT)和RP1976-18-6-4-2对孟加拉型的抗性与与PPO密切相关,570011对孟加拉型的抗性与POD活性密切相关。苗龄、光照强度和施氮量的单独作用或交互作用对不同水稻中防御酶活性有明显影响,但品种不同,其影响程度也不同。苗龄、光照强度和施氮量单独或是交互作用显著影响IR56中PAL和PPO、RHT和RP1976-18-6-4-2中PPO、Ptb33中POD及570011中PAL和POD的活性,苗龄显著影响IR56和RP1976-18-6-4-2中POD的活性,光照强度和施氮量单独或交互作用显著影响RHT和RP1976-18-6-4-2中PAL的活性,施氮量显著影响RHT中POD的活性,光照强度显著影响Ptb33中PAL的活性;苗龄和光照强度单独或交互作用显著影响570011中PPO的活性。