污水处理系统中厌氧氨氧化菌分布及影响因素

基于厌氧氨氧化的污水生物脱氮工艺近年来发展迅速,污水处理系统中厌氧氨氧化菌的分布和多样性成为了重要的研究方向。目前,在污水处理系统中曾检测出多种厌氧氨氧化(Anaerobic ammonium oxidation,Anammox)菌,最常被检测出的是待定布罗卡地菌Candidate Brocadia和待定斯图加特库氏菌Candidate Kuenenia的Anammox菌,并且研究发现单一生境下往往只存在一种类型的Anammox菌,但是影响Anammox菌分布和多样性的因素与机制目前仍不明确。系统总结了污水处理系统中,不同工艺形式和运行条件下的Anammox菌分布情况,归纳分析了关键因素对Anammox菌分布的影响,包括底物浓度和微生物比生长速率、污泥性质与微生物生境、多重因素的联合作用和影响等。在此基础上,阐述了Anammox菌分布机制研究的工程意义,并对该领域的研究方向和思路进行了展望。     

In situ morphology of nitrifying-like bacteria in aquaculture systems

[This corrects the article on p. 423 in vol. 31.].     

Quantitative role of shrimp fecal bacteria in organic matter fluxes in a recirculating shrimp aquaculture system

Microorganisms play integral roles in the cycling of carbon (C) and nitrogen (N) in recirculating aquaculture systems (RAS) for fish and shellfish production. We quantified the pathways of shrimp fecal bacterial activities and their role in C- and N-flux partitioning relevant to culturing Pacific white shrimp, Penaeus (Litopenaeus) vannamei, in RAS. Freshly produced feces from P. vannamei contained 0.6-7 × 10(10) bacteria g(-1) dry wt belonging to Bacteroidetes (7%), Alphaproteobacteria (4%), and, within the Gammaproteobacteria, almost exclusively to the genus Vibrio (61%). Because of partial disintegration of the feces (up to 27% within 12 h), the experimental seawater became inoculated with fecal bacteria. Bacteria grew rapidly in the feces and in the seawater, and exhibited high levels of aminopeptidase, chitinase, chitobiase, alkaline phosphatase, α- and β-glucosidase, and lipase activities. Moreover, fecal bacteria enriched the protein content of the feces within 12 h, potentially enriching the feces for the coprophagous shrimp. The bacterial turnover time was much faster in feces (1-10 h) than in mature RAS water (350 h). Thus, shrimp fecal bacteria not only inoculate RAS water but also contribute to bacterial abundance and productivity, and regulate system processes important for shrimp health.     

In situ morphology of nitrifying-like bacteria in aquaculture systems.

The in situ microbiota from several aquaculture facilities with active nitrification was examined by transmission electron microscopy of thin sections for the presence of bacteria that contained intracytoplasmic membranes characteristic of the nitrifying bacteria. Colonies of bacteria with the cellular morphology of a species of Nitrosomonas were found to be present in both the culture water and in the biological filter slime of a freshwater chinook salmon (Oncorhynchus tshawytscha) culture system. bacteria in the water possessed the normal nitrosomonas type of ultrastructure, whereas similar bacteria in the slime had an aberrant morphology due to multiple invaginations of the cell wall and cyto-membranes and a significantly greater number of ribosomes. These nitrosomonas-like bacteria lysed during enrichment in commonly used media. Bacteria with the morphology of species of Nitrosomonas and Nitrosococcus were also observed in colonies in the surface slimes of marine culture systems for striped bass (Morone saxatilis) and quahaug (Mercenaria mercenaria).     

Life cycle assessment of recirculating aquaculture systems: A case of Atlantic salmon farming in China

Recirculating aquaculture systems (RAS) are an alternative technology to tackle the major environmental challenges associated with conventional cage culture systems. In order to systematically assess the environmental performance of RAS farming, it is important to take the whole life cycle into account so as to avoid ad hoc and suboptimal environmental measures. So far, the application of life cycle assessment (LCA) in aquaculture, especially to indoor RAS, is still in progress. This study reports on an LCA of Atlantic salmon harvested at an indoor RAS farm in northern China. Results showed that 1 tonne live‐weight salmon production required 7,509 kWh farm‐level electricity and generated 16.7 tonnes of CO₂ equivalent (eq), 106 kg of SO₂ eq, 2.4 kg of P eq, and 108 kg of N eq (cradle‐to‐farm gate). In particular, farm‐level electricity use and feed product were identified as primary contributors to eight of nine impact categories assessed (54–95% in total), except the potential marine eutrophication (MEU) impact (dominated by the grow‐out effluents). Among feed ingredients (on a dry‐weight basis), chicken meal (5%) and krill meal (8%) dominated six and three, respectively, of the nine impact categories. Suggested environmental improvement measures for this indoor RAS farm included optimization of stocking density, feeding management, grow‐out effluent treatment, substitution of feed ingredients, and selection of electricity generation sources. In a generic context, this study can contribute to a better understanding of the life cycle environmental impacts of land‐based salmon RAS operations, as well as science‐based communication among stakeholders on more eco‐friendly farmed salmon.     

Anaerobic ammonium-oxidizing (anammox) bacteria and associated activity in fixed-film biofilters of a marine recirculating aquaculture system

Microbial communities in the biological filter and waste sludge compartments of a marine recirculating aquaculture system were examined to determine the presence and activity of anaerobic ammonium-oxidizing (anammox) bacteria. Community DNA was extracted from aerobic and anaerobic fixed-film biofilters and the anaerobic sludge waste collection tank and was analyzed by amplifying 16S rRNA genes by PCR using anammox-selective and universal GC-clamped primers. Separation of amplified PCR products by denaturing gradient gel electrophoresis and sequencing of the different phylotypes revealed a diverse biofilter microbial community. While Planctomycetales were found in all three communities, the anaerobic denitrifying biofilters contained one clone that exhibited high levels of sequence similarity to known anammox bacteria. Fluorescence in situ hybridization studies using an anammox-specific probe confirmed the presence of anammox Planctomycetales in the microbial biofilm from the denitrifying biofilters, and anammox activity was observed in these biofilters, as detected by the ability to simultaneously consume ammonia and nitrite. To our knowledge, this is the first identification of anammox-related sequences in a marine recirculating aquaculture filtration system, and our findings provide a foundation for incorporating this important pathway for complete nitrogen removal in such systems.     

Mass production of nitrifying bacterial consortia for the rapid establishment of nitrification in saline recirculating aquaculture systems

Two distinct nitrifying bacterial consortia, namely an ammonia oxidizing non-penaeid culture (AMONPCU-1) and an ammonia oxidizing penaeid culture (AMOPCU-1), have been mass produced in a nitrifying bacterial consortia production unit (NBCPU). The consortia, maintained at 4°C were activated and cultured in a 2 l fermentor initially. At this stage the net biomass (0.105 and 0.112 g/l), maximum specific growth rate (0.112 and 0.105/h) and yield coefficients (1.315 and 2.08) were calculated respectively, for AMONPCU-1 and AMOPCU-1 on attaining stationary growth phase. Subsequently on mass production in a 200 l NBCPU under optimized culture conditions, the total amounts of NH₄ ⁺–N removed by AMONPCU-1 and AMOPCU-1 were 1.948 and 1.242 g/l within 160 and 270 days, respectively. Total alkalinity reduction of 11.7–14.4 and 7.5–9.1 g/l were observed which led to the consumption of 78 and 62 g Na₂CO₃. The yield coefficient and biomass of AMONPCU-1 were 0.67 and 125.3 g/l and those of AMOPCU-1 were 1.23 and 165 g/l. The higher yield coefficient and growth rate of AMOPCU-1 suggest better energy conversion efficiency and higher CO₂ fixation potential. Both of the consortia were dominated by Nitrosomonas-like organisms. The consortia may find application in the establishment of nitrification within marine and brackish water culture systems.     

封闭循环养殖系统中β变形菌亚纲氨氧化细菌的引物特异性

摘要:【目的】为探讨4 对不同的引物对封闭循环养殖系统生物膜中β 变形菌亚纲氨氧化细菌(β-AOB) 的特异性差异。【方法】采用16S rDNA 文库克隆技术对β-AOB 的多样性进行了分析。【结果】以引物CTO189f/CTO654r 扩增构建的文库中所含β-AOB 比例最高,可达67. 3%。不同封闭循环养殖系统的生物膜对引物的扩增效率有明显的影响,其中以养殖尼罗罗非鱼的封闭循环养殖系统生物膜为DNA 来源的,引物均有较高的扩增效率。【结论】针对封闭循环养殖系统生物膜中的β-AOB,特异性最高的是CTO189f /CTO654r 引物。     

Anammox bacteria: from discovery to application

Anaerobic ammonium oxidation (anammox) bacteria, which were discovered in waste-water sludge in the early 1990s, have the unique metabolic ability to combine ammonium and nitrite or nitrate to form nitrogen gas. This discovery led to the realization that a substantial part of the enormous nitrogen losses that are observed in the marine environment--up to 50% of the total nitrogen turnover--were due to the activity of these bacteria. In this Timeline, Gijs Kuenen recalls the discovery of these unique microorganisms and describes the continuing elucidation of their roles in environmental and industrial microbiology.     

Characterizing the structural diversity of a bacterial community associated with filter materials in recirculating aquaculture systems of Scortum barcoo

The bacterial community structure associated with filter materials in the recirculating aquaculture system of Scortum barcoo was investigated using the 16S rRNA gene clone library method. Preliminary results showed that the clone library constructed from the initial operation condition was characterized by 31 taxa of bacteria belonging to eight phyla including Proteobacteria, Acidobacteria, Firmicutes, Fusobacteria, Sphingobacteria, Bacteroidetes, Verrucomicrobiae, and Actinobacteria. There were 14 taxa of bacteria belonging to four phyla including Proteobacteria, Acidobacteria, Planctomycetacia, and Nitrospirae from the stable operation condition where the water quality was well maintained. Nitrospirae was only found under the stable operation condition in this study. Our results further indicated that Nitrospira was dominated by members of the Nitrospira sp. lineages, with a minor fraction related to Nitrospira moscoviensis and an unknown Nitrospira cluster. These great differences of both diversity and composition between two operation conditions suggested that the composition of the microbial community varied with the degree of water quality in the recirculating aquaculture system of S.?barcoo.     

Mechanical mode floating medium filters for recirculating systems in aquaculture for higher solids retention and lower freshwater usage

The aim of this work was to develop a better understanding of a floating medium in a mechanical filtration mode. The experiments were carried out using a commonly available polystyrene floating medium filter with the grain size of 1mm. A sand medium filter with the similar grain size was also tested for the comparison. A short-term trial of 2h and a long-term of 20 days filtration times were conducted with three custom manufactured pressurized filters of 16l. The filters were operated under three different configurations: (i) upflow with floating media (UFMF), (ii) downflow with floating media (DFMF) and (iii) downflow with a sand medium (DSF). The results of the long-term trial indicated that at a flow rate of 22 m/h, the UFMF and DSF had similar solid removal capacity with an average total suspended solids (TSS) removal efficiency of 60%. The DFMF could only remove 33% of TSS. However, during the short-term trial, TSS removal efficiency of the UFMF was better compared to the DSF (e.g., 71%, 56% and 57% of TSS removal in UFMF compared to 66%, 49% and 41% in the DSFF at the flow rates of 20, 25 and 31m/h, respectively). The energy requirements of each filter were compared by measuring the pressure differential across each filter. The long-term trial indicated that the UFMF had a significantly less pressure differential (44 kPa) compared to the DSF (80 kPa) (p<0.001). This was further confirmed that at different flow rates whereby the DSF displayed higher pressure differentials for filtration rates at 350, 450, 550 and 800 l/h. The study indicated that floating medium filter was better and more applicable to recirculating aquaculture systems than conventional pressurized sand filter.     

Inhibitory effects of free ammonia on Anammox bacteria

Anammox bacteria can effectively treat high ammonia and nitrite concentrations under anoxic environments. However, the presence of high ammonia and nitrite concentrations may cause free ammonia and nitrous acid inhibition at high pH and temperature environments. In this study, the inhibitory effect of free ammonia on Anammox bacteria was investigated in a lab-scale upflow fixed-bed reactor with Kaldnes biofilm carriers. Results of continuous operation showed that inhibition was not observed in the Anammox reactor when the free ammonia concentration gradually increased up to 150?mg/L. However, Anammox activity suddenly dropped to 10 % when the free ammonia concentration reached to 190?mg/L. Nevertheless, high influent ammonia and nitrite concentrations up to 1,500 and 500?mg/L, respectively, did not noticeably inhibit the Anammox activity. Gradually decreasing Anammox activity was also supported by fluorescent in situ hybridization (FISH) analysis. FISH and 16S rRNA gene analysis results revealed that main Anammox organisms were phylogenetically related to Candidatus Kuenenia stuttgartiensis, Candidatus Jettenia asiatica and Candidatus Brocadia anammoxidans.     

循环海水养鱼系统中pH值调节技术研究

分别采用NaHCO3、Na2CO3和NaOH 3种碱性试剂,3种添加方式调节循环海水养殖系统中水样的pH值。结果表明:从NaHCO3、Na2CO3和NaOH相应的滴定曲线确定突跃点pH值分别为7.25±0.04,7.22±0.01,7.11±0.01;发现添加714m g/L的NaHCO3可以有效提高并稳定水体的pH值,调节效果优于另两种碱试剂;"一次性"、"分四次"和"连续性"3种添加方式中,以"连续性"添加的水体pH值上升最平缓。要使循环海水的pH值维持在7.20～8.00之间,应在突跃点pH值7.22前夕,用连续添加的方式加入NaHCO3试剂,周期约为15～19 d。     

青石斑鱼海水循环水养殖水体的细菌群落特征北大核心CSCD

【目的】为揭示工厂化循环水青石斑鱼养殖水体的细菌群落特征,比较患病养殖池与健康养殖池的细菌群落结构差异,探讨细菌群落结构与青石斑鱼病害相关的相关性。【方法】采用Illumina Miseq高通量测序方法,分析比较了患病和健康养殖水体细菌群落结构、α-多样性指数(包括多度、均一度和系统发育多样性);并结合传统方法从患病青石斑鱼病灶部位分离疑似病原菌。【结果】患病和健康养殖水体中细菌群落的α-多样性并无太大差异,但主坐标分析与热图样本聚类分析表明细菌群落结构明显不同。二者的优势细菌门均为Proteobacteria、Verrucomicrobia和Bacteroidetes,但它们的相对丰度差异显著。患病养殖水体中α-Proteobacteria(25.07%)和γ-Protebacteria(22.74%)丰度相当,而健康养殖水体中γ-Protebacteria(40.49%)显著高于α-Proteobacteria(10.87%)。患病水体的Verrucomicrobia丰度(26.4%)远高于健康水体(10.9%);而Bacteroidetes的相对丰度则相反(12.3%vs 20.9%)。主要的差异类群包括α-Proteobacteria的Rhodobacteraceae和Rhodospirillaceae,γ-Proteobacteria的Alteromonadaceae、HTCC2188和Oceanospirillaceae,Verrucomicrobia的Verrucomicrobiaceae和Bacteroidetes的Cryomorphaceae。更表现在核心微生物类群的差异,健康养殖池水体以Glaciecola、HTCC、Sediminicola、Prevotella等对于养殖动物有益或无害的属为核心微生物;而患病养殖池水体则以Vibrio、Rubritalea、Oleibacter等病原菌或对养殖动物不利的属为核心微生物。从患病青石斑鱼的皮肤、肝脏和脾脏共分离得到弧菌20株,Acinetobacter haemolyticus 1株。【结论】患病的青石斑鱼循环水养殖水体中的细菌群落明显不同于健康养殖水体,特别是核心微生物的差异,其以Vibrio等病原菌或对养殖动物不利的属为主。该结果为青石斑鱼循环水养殖系统的管理、病害的诊断和监测提供理论与实验基础。     

以聚羟基丁酸戊酸共聚酯为碳源去除循环水养殖系统的硝酸盐及生物膜中微生物群落动态

【目的】利用聚羟基丁酸戊酸共聚酯(PHBV)作为固体碳源和生物膜载体,去除循环水养殖系统的硝酸盐,并研究固体碳源反硝化反应器不同运行阶段生物膜中微生物群落结构的动态变化。【方法】采用PCR-DGGE技术对反硝化反应器中微生物群落结构的动态变化进行了分析,采用传统纯培养方法分离反应器中降解PHBV的细菌。【结果】连接固相反硝化反应器能使循环水养殖系统中积累的硝酸盐显著降低,并维持在较低水平(小于10 mg/L),而常规循环水养殖系统中硝酸盐浓度持续增加。系统发育树聚类分析结果表明,反硝化反应器生物膜细菌归属于变形菌门(β-proteobacteria、γ-proteobacteria和δ-proteobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)。反应器运行初期(40 d)的优势种群主要为Acidovorax和Bacillus;运行后期(150 d)的优势种群依次为Clostridium、Desulfitobacterium、Dechloromonas、Pseudoxanthomonas和Flavobacterium。从反应器中分离到的PHBV降解菌株分别归属于Acidovorax、Methylibium、Pseudoxanthomonas和Dechloromonas。【结论】利用PHBV为碳源能有效去除循环水养殖系统的硝酸盐。明确了反硝化反应器在运行过程中,碳源表面生物膜的优势菌群及其动态变化。     

Anammox bacterial populations in deep marine hypersaline gradient systems

To extend the knowledge of anaerobic ammonium oxidation (anammox) habitats, bacterial communities were examined in two hypersaline sulphidic basins in Eastern Mediterranean Sea. The 2 m thick seawater–brine haloclines of the deep anoxic hypersaline basins Bannock and L’Atalante were sampled in intervals of 10 cm with increasing salinity. ¹⁵N isotope pairing incubation experiments showed the production of ²⁹N₂ and ³⁰N₂ gases in the chemoclines, ranging from 6.0 to 9.2 % salinity of the L’Atalante basin. Potential anammox rates ranged from 2.52 to 49.65 nmol N₂ L^?1 day^?1 while denitrification was a major N₂ production pathway, accounting for more than 85.5 % of total N₂ production. Anammox-related 16S rRNA genes were detected along the L’Atalante and Bannock haloclines up to 24 % salinity, and the amplification of the hydrazine synthase genes (hzsA) further confirmed the presence of anammox bacteria in Bannock. Fluorescence in situ hybridisation and sequence analysis of 16S rRNA genes identified representatives of the marine anammox genus ‘Candidatus Scalindua’ and putatively new operational taxonomic units closely affiliated to sequences retrieved in marine environments that have documented anammox activity. ‘Scalindua brodae’ like sequences constituted up to 84.4 % of the sequences retrieved from Bannock. The anammox community in L’Atalante was different than in Bannock and was stratified according to salinity increase. This study putatively extends anammox bacterial habitats to extremely saline sulphidic ecosystems.     

Differential cytokine response from dendritic cells to commensal and pathogenic bacteria in different lymphoid compartments in humans

Resident host microflora condition and prime the immune system. However, systemic and mucosal immune responses to bacteria may be divergent. Our aim was to compare, in vitro, cytokine production by human mononuclear and dendritic cells (DCs) from mesenteric lymph nodes (MLNs) and peripheral blood mononuclear cells (PBMCs) to defined microbial stimuli. Mononuclear cells and DCs isolated from the MLN (n = 10) and peripheral blood (n = 12) of patients with active colitis were incubated in vitro with the probiotic bacteria Lactobacillus salivarius UCC118 or Bifidobacterium infantis 35624 or the pathogenic organism Salmonella typhimurium UK1. Interleukin (IL)-12, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, and IL-10 cytokine levels were quantified by ELISA. PBMCs and PBMC-derived DCs secreted TNF-alpha in response to the Lactobacillus, Bifidobacteria, and Salmonella strains, whereas MLN cells and MLN-derived DCs secreted TNF-alpha only in response to Salmonella challenge. Cells from the systemic compartment secreted IL-12 after coincubation with Salmonella or Lactobacilli, whereas MLN-derived cells produced IL-12 only in response to Salmonella. PBMCs secreted IL-10 in response to the Bifidobacterium strain but not in response to the Lactobacillus or Salmonella strain. However, MLN cells secreted IL-10 in response to Bifidobacteria and Lactobacilli but not in response to Salmonella. In conclusion, commensal bacteria induced regulatory cytokine production by MLN cells, whereas pathogenic bacteria induce T cell helper 1-polarizing cytokines. Commensal-pathogen divergence in cytokine responses is more marked in cells isolated from the mucosal immune system compared with PBMCs.