有机碳对混合菌群亚硝酸盐氮氧化的影响

自养硝化过程在自然界氮素循环和污水处理系统脱氮过程中起着关键作用。因此，了解有机碳对硝化的影响和硝化菌与异养菌之间的竞争对微生物生态学和污水处理系统设计都很重要。目前对氨氧化到硝酸盐氮过程的研究文献很多，但对亚硝酸盐氧化过程在异养菌的存在下如何受到有机碳影响的研究甚少。本文从生理生化指标、基因组学、蛋白组学三方面考察了在实验室条件下有机碳（乙酸钠）对硝化细菌和异养菌组成的混合菌群的硝化性能、菌群结构及代谢功能的变化的影响。 全文分为两大部分： 第一部分为乙酸钠对游离态硝化混合菌群的硝化性能和菌群结构的短期影响。混合菌株先在自养条件下进行连续培养，两个月后硝化速率达到20 mg N/(L·d)；而后离心收集菌体进行批式实验。在批式反应器中，初始亚硝氮均为126mg N/ L，乙酸钠-C 与亚硝酸盐-N 的比分别为0，0.44，0.88，4.41，8.82。结果表明：在低C/N 比（0.44 和0.88）时，亚硝酸盐去除速率比C/N=0 下高，细菌呈现一次生长；而在高C/N 比（4.41 和8.82）时，出现连续的硝化反硝化，亚硝酸盐去除率仍比对照下高，细菌呈现二次生长。不同C/N 比下微生物群落明显不同，优势菌群从自养和寡营养细菌体系（包括亚硝酸盐氧化菌，拟杆菌门,α-变形菌纲,浮霉菌门和绿色非硫细菌下的一些菌株）过渡到异养和反硝化菌体系 （γ-变形菌纲的菌株尤其是反硝化菌Pseudomonas stutzeri 和P. nitroreducens 占主导）。 第二部分为乙酸钠对硝化混合菌群生物膜的硝化性能和菌群结构的长期影响。接种富集的硝化混合菌群于装有组合式填料的三角瓶中，于摇床中自养培养；两个月后填料上形成生物膜的硝化速率达到20 mg N/ (L·d)；而后进行长期实验，每12 小时更换混合营养培养基（亚硝氮约200 mg N/ L，C/N 比同上）。结果显示：相较于C/N 比=0 时的亚硝酸盐氧化反应来说，低C/N 比出现了部分的反硝化，而高C/N 比则是几乎完全的反硝化。与对照比，C/N=0.44 时亚硝酸盐氧化速率并未受乙酸钠的影响，反而上升了，但C/N=0.88 时亚硝酸盐氧化速率有所下降。菌群结构分析表明自养对照与混合营养下微生物群落的不同；PCR-DGGE未检测出混合营养下硝化杆菌的存在，而显示异养菌尤其是反硝化菌的大量存 在。荧光定量PCR 结果表明随C/N 比上升，硝化杆菌数量从2.42 × 104 下降到1.34× 103 16S rRNA gene copies/ ng DNA，反硝化菌由0 增加至2.51 × 104 nosZgene copies/ ng DNA。SDS-PAGE 的结果表明不同C/N 比下的蛋白组较为复杂且呈现一定的差异性。 有机碳对亚硝氮氧化及微生物群落的影响很复杂，本文分别讨论了对游离态和生物膜固定态两种状态的混合菌群相应的短期和长期影响研究。研究发现，有机碳并非一定带来硝化的负影响，如果控制在适当的C/N 比范围，有机碳是有利于亚硝氮氧化的。这些发现阐明了有机碳和硝化反硝化的关系，填补了硝化微生物生态学上的空白，对污水处理系统中减少异养菌的影响并提高氮去除率有一定理论指导意义。 Nitrification plays a key role in the biological removal of nitrogen in both nature and wastewater treatment plant (WWTP). So, understanding of the effect of organic carbon on nitrification and the competition between nitrifying bacteria and heterotrophic bacteria is important for both microbial ecology and WWTP design and operation. Despite the fact that the nitrification process of ammonia to nitrate has been extensively investigated, it is not known how the process of nitrite oxidization is affected by organic carbon when heterotrophic bacteria are present. By measuring different physiological and biochemical parameters, as well as using genomic DNA and proteome analysis, we investigated the influence of organic (acetate) on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria under laboratory conditions. The dissertation involves two parts: Part one deals with the effect of organic matter on functional performance and bacterial community shift of nitrite-oxidizing and heterotrophic bacteria under suspended state. The bacteria were prepared in a continuous-flow stirred reactor under autotrophic condition; after two months, the nitrification rate of the culture reached about 20 mg N/ (L·d); then the bacteria were harvested for the next batch experiments. The initial concentrations of nitrite were 126 ± 6 mg N/ L in all flasks, and sodium acetate (C) to nitrite (N) ratios were 0, 0.44, 0.88, 4.41, and 8.82, respectively. The results showed that at low C/N ratios (0.44 or 0.88), the nitrite removal rate was higher than that obtained under autotrophic condition and the bacteria had single growth phase, while at high C/N ratios (4.41 or 8.82), continuous aerobic nitrification and denitrification occurred besides higher nitrite removal rates, and the bacteria had double growth phases. The community structure of total bacteria strikingly varied with the different C/N ratios; the dominant populations shifted from autotrophic and oligotrophic bacteria (NOB, and some strains of Bacteroidetes, Alphaproteobacteria, Actinobacteria, and green nonsulfur bacteria) to heterotrophic and denitrifying bacteria (strains of Gammaproteobacteria, especially Pseudomonas stutzeri and P. nitroreducens). Part two describes the influence of acetate on nitrite oxidizing performance, community structure and metabolic function of nitrite-oxidizing and heterotrophic bacteria in biofilms. Bacterial enrichments was transferred into flasks with polypropylene carriers and cultured under agitated and autotrophic condition. After two month, the biofilms grown on the carriers had a nitrification rate of about 20 mg N/ (L·h); then the biofilms were refreshed with mixotrophic medium (nitrite were 200 mg N/ L in all flasks, and C/N ratios was the same as above) every 12 h. the results show: normal nitrite oxidization reactions were performed when C/N = 0, but nitrite oxidization and partial denitrification occurred with low C/N ratios (0.44 or 0.88). At high C/N ratios (4.41 or 8.82), we mainly observed denitrification. In contrast to C/N = 0, the nitrite oxidization rate was unaffected when C/N = 0.44, but decreased with C/N = 0.88. The structure of bacterial communities varied significantly between autotrophic and mixotrophic conditions. Nitrobacter was hard to detect by PCR-DGGE while heterotrophs and especially denitrifiers were in the majority under mixotrophic conditions. Real-time PCR indicated that the Nitrobacter population decreased from 2.42 × 104 to 1.34 × 103 16S rRNA gene copies/ ng DNA, while the quantity of denitrifiers obviously increased from 0 to 2.51×104 nosZ gene copies/ ng DNA with an increasing C/N ratio. SDS-PAGE indicated the complexity of and a certain difference between the proteome of nitrite-oxidizing and heterotrophic bacteria at different C/N ratios. We conclude that the influence of organic matter on nitrite oxidation and the community structure of NOB and heterotrophic bacteria is complex. In this dissertation, we focused on how sodium acetate influenced the system both under suspended state and in biofilms. We observed that acetate did not necessarily have a negative impact on nitrification. Instead, an appropriate amount of acetate benefited both nitrite oxidization and denitrification. These findings provide a greater understanding about the relationship between organics and nitrification; they fill the gaps in the field of microbial ecology of nitrifying bacteria; they also provide insight into how to minimize the negative impact of heterotrophic bacteria and maximize the benefit of nitrogen removal in biological treatment systems.

ISOLATION AND CHARACTERIZATION OF CHLORELLA SOROKINIANA GXNN01 (CHLOROPHYTA) WITH THE PROPERTIES OF HETEROTROPHIC AND MICROAEROBIC GROWTH

Heterotrophic and anaerobic microalgae are of significance in both basic research and industrial application. A microalga strain was isolated from a wastewater treatment pond and identified as Chlorella sorokiniana Shihira et W. R. Krauss GXNN01 in terms of morphology, physiology, and phylogeny. The strain grows rapidly in heterotrophic or mixotrophic conditions with addition of various carbon sources, and even in anaerobic conditions. The maximum growth rate reached 0.28 d(-1) when using D,L-malate as the carbon source, and the protein content of the microalgae was 75.32% in cell dry weight. The strain was shown to be capable of (1) utilizing D, L-malate only with light, (2) inhibiting photosynthesis in mixotrophic growth, and (3) growing in anaerobic conditions with regular photosynthesis and producing oxygen internally. This study demonstrates the influence of oxygen (aerobic vs. anaerobic) and metabolic regime (autotrophy, mixotrophy, heterotrophy) on the physiological state of the cell.

Extracellular hydrolytic enzyme screening of culturable heterotrophic bacteria from deep-sea sediments of the Southern Okinawa Trough

The Southern Okinawa Trough is an area of focused sedimentation due to particulate matter export from the shelf of the East China Sea and the island of Taiwan. In order to understand the geomicrobiological characteristics of this unique sedimentary environment, bacterial cultivations were carried out for an 8.61 m CASQ core sediment sample. A total of 98 heterotrophic bacterial isolates were characterized based on 16S rRNA gene phylogenetic analysis. These isolates can be grouped into four bacterial divisions, including 13 genera and more than 20 species. Bacteria of the gamma-Proteobacteria lineage, especially those from the Halomonas ( 27 isolates) and Psychrobacter ( 20 isolates) groups, dominate in the culturable bacteria assemblage. They also have the broadest distribution along the depth of the sediment. More than 72.4% of the isolates showed extracellular hydrolytic enzyme activities, such as amylases, proteases, lipases and Dnases, and nearly 59.2% were cold-adapted exoenzyme-producers. Several Halomonas strains show almost all the tested hydrolases activities. The wide distribution of exoenzyme activities in the isolates may indicate their important ecological role of element biogeochemical cycling in the studied deep-sea sedimentary environment.

A bioaugmentation failure caused by phage infection and weak biofilm formation ability

Two biological aerated filters (BAF) were setup for ammonia removal treatment of the circulation water in a marine aquaculture. One of the BAFs was bioaugmented with a heterotrophic nitrifying bacterium, Lutimonas sp. H10, where the ammonia removal was not improved and the massive inoculation was even followed by a nitrification breakdown from day 9 to 18. The nitrification was remained stable in control BAF operated under the same conditions. Fluorescent in situ hybridization (FISH) with rRNA-targeted probes and cultivable method revealed that Lutimonas sp. H10 almost disappeared from the bioaugomented BAF within 3 d, and this was mainly due to the infection of a specific phage as revealed by flask experiment, plaque assay and transmission electron observation. Analyses of 16S rRNA gene libraries showed that bacterial groups from two reactors evolved differently and an overgrowth of protozoa was observed in the bioaugmented BAR Therefore, phage infection and poor biofilm forming ability of the inoculated strain are the main reasons for bioaugmentation failure. In addition, gazing by protozoa of the bacteria might be the reason for the nitrification breakdown in bioaugmented BAF during day 9-18.

Spatial variability in biomass and production of heterotrophic bacteria in the East China Sea and the Yellow Sea

The distributions of heterotrophic bacterial abundance and production were investigated in the East China Sea and the Yellow Sea during the autumn of 2000 and spring of 2001. Bacterial abundance varied in the range 3.2-15.7 (averaging 5.7) x 10(5) and 2.3-13.6 (averaging 6.2) x 10(5) cells cm(-3) in the spring and autumn, respectively. During autumn, bacterial production (BP) (0.27-7.77 mg C m(-3) day(-1)) was on average 3 fold that in spring (0.001-2.04 mg C m(-3) day(-1)). Bacterial average turnover rate (ratio of bacterial production:bacterial biomass, mu=0.21 day(-1)) in autumn was 3 times as high as in spring (0.07 day(-1)). The ratio of integrated bacterial biomass to integrated phytoplankton biomass in the euphotic zone ranged from 4 to 101% (averaging 35%) in spring and 24 to 556% (averaging 121%) in autumn. The results indicate that the distributions of heterotrophic bacteria were controlled generally by temperature in spring and additionally by substrate supply in autumn. (C) 2010 Elsevier Ltd. All rights reserved.

Spatial variations of size-fractionated chlorophyll, cyanobacteria and heterotrophic bacteria in the Central and Western Pacific

Geographic and vertical variations of size-fractionated (0.2-1 mu m, 1-10 mu m, and >10 mu m) Chlorophyll a (Chl.a) concentration, cyanobacteria abundance and heterotrophic bacteria abundance were investigated at 13 stations from 4 degrees S, 160 degrees W to 30 degrees N, 140 degrees E in November 1993. The results indicated a geographic distribution pattern of these parameters with instances of high values occurring in the equatorial region and offshore areas, and with instance of low values occurring in the oligotrophic regions where nutrients were almost undetectable. Cyanobacteria showed the highest geographic variation (ranging from 27x10(3) to 16,582x10(3) cell l(-1)), followed by Chl.a (ranging from 0.048 to 0.178 mu g l(-1)), and heterotrophic bacteria (ranging from 2.84x10(3) to 6.50 x 10(5) cell l(-1)). Positive correlations were observed between nutrients and Chl.a abundance. Correspondences of cyanobacteria and heterotrophic bacteria abundances to nutrients were less significant than that of Chl.a. The total Chl.a was accounted for 1.0-30.9%, 35.9-53.7%, and 28.1-57.3% by the >10 mu m, 1-10 mu m and 0.2-1 mu m fractions respectively. Correlation between size-fractionated Chl.a and nutrients suggest that the larger the cell size, the more nutrient-dependent growth and production of the organism. The ratio of pheophytin to chlorophyll implys that more than half of the > 10 mu m and about one third of the 1-10 mu m pigment-containing particles in the oligotrophic region were non-living fragments, while most of the 1-10 mu m fraction was living cells. In the depth profiles, cyanobacteria were distributed mainly in the surface layer, whereas heterotrophic bacteria were abundant from surface to below the euphotic zone. Chl.a peaked at the surface layer (0-20 m) in the equatorial area and at the nitracline (75-100 m) in the oligotrophic regions. Cyanobacteria were not the principle component of the picoplankton. The carbon biomass ratio of heterotroph to phytoplankton was greater than 1 in the eutrophic area and lower than 1 in oligotrophic waters.

北京东灵山暖温带典型植被类型物种构成及物种多样性对氮素转化过程的影响

通过调查东灵山地区7个典型植被类型的物种多样性，并测定各林型中植物叶片养分特征，凋落物品质，土壤理化特性，豆科植物的生物固氮作用以及土壤有机质的净N矿化与硝化作用，研究了不同林型及其物种多样性对氮素转化过程的可能影响。结果表明： 1) 各林型乔灌草三层13个物种多样性指数之间都不同程度存在差异，且都表现基本一致的大小顺序，各样地乔灌草三层3个丰富度指数和5个多样性指数之间大都表现草本层>灌木层>乔木层的大小顺序。 2) 林型对植物叶片养分特征、叶片凋落物品质指标、土壤的大多数理化特性都有显著影响，其中，落叶阔叶林与针叶林之间的差异最为明显，优势种不同的针叶林之间也不同程度地存在差异。各林型物种多样性对植物叶片养分特征，凋落物品质和土壤理化特性也都不同程度存在显著影响，且乔灌草三层的物种多样性的影响不同。我们的结果支持物种多样性对生态系统过程存在显著影响的观点。 3) 胡枝子(Lespedeza bicolor)％FNDA值较低，在0～53.3％之间，与国外学者对某些木本豆科固氮植物的研究结果接近，但低于三叶草、野豌豆等草本豆科固氮植物。胡枝子生物固氮具有明显的季节差异，不同样地生境对胡枝子％FNDA值也存在显著影响，大都都表现铁塔辽东栎样地>重力点样地>站下灌丛样地>垭口样地的趋势。除胡枝子之外的其他豆科植物的％FNDA都高于胡枝子，且存在种间差异，以三籽两型豆(Amphicarpaea capillipes)最高(100％)，歪头菜(Vicia unijuga)最低(平均66.4％)，这些结果与国外相关研究接近。不同林型之间和同一林型不同样地之间也影响豆科植物的％FNDA。东灵山地区有相当数量的植物种具有和固氮植物相近的δ15N值和全N含量，具有潜在的生物固氮能力。6个林型中所有豆科植物的％FNDA平均值主要受乔木层和草本层物种多样性的影响，乔木层和草本层物种多样性提高，豆科植物的％FNDA将随之显著降低。 4) 气温、土壤温度和降水量显著影响各林型净氮硝化速率的季节动态，都表现为温度升高，降水量增加，净氮硝化速率也随之增大。 5) 在不同取样时间，不同林型在土壤NH4+、N03-含量，矿化、硝化速率以及年度总净矿化、硝化量之间均不同程度存在显著差异，其中，以辽东栎落叶阔叶林与针叶林之间，常绿针叶林与落叶针叶林之间，纯林与混交林之间的差异最为明显。 6) 土壤初始N0_3含量与年度总净硝化量，年度总净矿化量及其占TKN的百分比呈显著正相关关系;植物叶片全N浓度与净氮矿化、硝化作用呈显著负相关：植物叶片凋落物的品质指标与净氮矿化、硝化作用没有显著相关。 7) 胡枝子固氮作用(％FNDA值)越强，土壤有机氮的净硝化量就越大，胡枝子在从大气中获取大量N，的同时，很可能会增加氮素的淋溶损失量。胡枝子固氮作用(％FNDA值)与净氮矿化量不存在显著相关关系。 8) 只有乔木层和草本层部分种多样性指数对净氮矿化、硝化量存在显著影响。乔木层均匀度提高，土壤净氮碳化量将随之增大;而草本层物种多样性提高，均匀度提高，净氮矿化、硝化作用将随之降低，草本层植物对土壤氮素矿化作用具有显著抑制作用。 9) 落叶阔叶林与针叶林的供氮能力和维持无机氮素的能力之间存在比较明显的差异，而不同针叶林的矿化／硝化作用也有所差别。其中，辽东栎落叶阔叶林的供氮能力和维持氮素能力均高于针叶林和山杏灌丛;油松林的供氮能力与防止氮素损失的能力显然要强于落叶松林和山杏灌丛。 10) 尽管箭叶锦鸡儿灌丛植物叶片与凋落物中全N、全P浓度在大多数取样点上都低于硕桦(高于草甸)，但表层土中全N浓度高于硕桦和草甸，且其土壤有机质的供氮能力以及维持氮素能力都高于硕桦林和草甸，表明，锦鸡儿灌丛为侵入草甸和硕桦入侵提供了良好的养分条件，在该演替序列的发展过程中起了一定的推动作用。

暖温带典型森林生态系统未来碳平衡的LPJ-GUESS模型模拟与分析

森林作为陆地生态系统的主体，在全球陆地碳循环中起着决定性作用。实测和模型研究均表明北半球的森林是重要的大气CO2汇，在缓解全球碳收支失衡中发挥着关键作用。过去几十年北半球所经历的显著气候变化，已经很大地改变了陆地生态系统的碳平衡状况。随着未来100年气候变化继续增大，对未来气候变化下森林生态系统碳平衡的预测研究就尤为重要。 北京山区森林属于典型的暖温带森林生态系统，前人对本区森林的植被特征、生态系统结构和功能、养分循环以及长期动态变化等都进行了深入的研究。然而长期的人类活动已使本区原生的地带性植被破坏殆尽。因此，对该区域森林生态系统碳平衡的模拟研究可以帮助我们认识其生态系统碳平衡变化特点及未来气候变化对其潜在的影响。 本研究采用LPJ-GUESS植被动态机理模型，利用IPCC于2000年发布的《排放情景特别报告》(SRES)的A2和B2两种情景下不同气候模式对华北地区未来100年温度和降水预测的平均值以及相应大气CO2浓度变化情景进行驱动，模拟北京山区未来100年暖温带森林生态系统的净初级生产力和碳平衡，尽可能真实地反映未来百年的变化趋势。通过比较当前和未来气候情景下北京山区以辽东栎（Quercus liaotungensis）为优势种的落叶阔叶林、以白桦（Betula platyphylla）为主的落叶阔叶林和油松（Pinus tabulaeformis）为主的针阔混交林三种典型暖温带森林生态系统的碳平衡差异，了解未来北京山区这三种暖温带森林生态系统的碳源汇功能，认识气候变化和大气CO2浓度升高对净初级生产力（Net primary productivity, NPP）、净生态系统碳交换（Net ecosystem exchange, NEE）、土壤异养呼吸（Heterotrophic respiration, Rh）和碳储量（Carbon biomass, C biomass）的影响，以及不同生态系统碳平衡对气候变化响应的异质性。 结果表明，未来100年两种气候情景下三种森林生态系统的NPP和Rh均增加，并且A2情景下增加的程度更大。由于三种生态系统树种组成的不同，未来气候情景下各自NPP和Rh增加的比例不同，导致三者NEE的变化也相异：100年后辽东栎林由碳汇转变为弱碳源，白桦林仍保持为碳汇但功能减弱，油松林成为一个更大的碳汇。三种森林生态系统的碳生物量在未来气候情景下均增大，21世纪末与20世纪末相比：辽东栎林在A2情景下碳生物量增加的比例为27.6%，大于B2情景下的19.3%；白桦林和油松林在B2情景下碳生物量增加的比例分别为34.2%和52.2%，大于A2情景下的30.8%和28.4%。各森林类型碳平衡状况不同，原因除气候因素外，主要是由于树种组成的差异所导致。SRES A2和B2两种气候情景相比，相对较低的排放情景（B2）下，生态系统有更高的碳储量。

Extracellular phosphatases produced by phytoplankton and other sources in shallow eutrophic lakes (Wuhan, China): taxon-specific versus bulk activity

Extracellular phosphatases are an important part of the phosphorus cycle in aquatic environments. Phosphatase activity (PA) in plankton was studied in seven subtropical shallow lakes of different exploitation management and trophic status in the urban area of Wuhan City. Bulk PA was rather high (range 1.1-11 mu mol l(-1) h(-1)), although concentrations of soluble reactive phosphorus (SRP) were also high (range 27 mu g P l(-1) to similar to 1.5 mg P l(-1)) in all lakes. Cell-associated extracellular PA in phytoplankton was detected using the fluorescence-labelled enzyme activity technique. Phytoplankton species partly contributed to the bulk PA. We found explicit differences in the presence of cell-associated phosphatase within the main phytoplankton groups; species belonging to Chlorophyta and Dinophyta were regularly phosphatase-positive, while Cyanophyta and Bacillariophyceae were phosphatase-negative in all but one case. Furthermore, there is a certain potential of extracellular phosphatases produced by heterotrophic nanoflagellates in most of the lakes. This new finding compromises the 'traditional' interpretation of bulk phosphatase data as being due to overall phytoplankton or bacterial P regeneration.

Depletion of Vipp1 in Synechocystis sp PCC 6803 affects photosynthetic activity before the loss of thylakoid membranes

A vipp1 mutant of Synechocystis sp. PCC 6803 could not be completely segregated under either mixotrophic or heterotrophic conditions. A vipp1 gene with a copper-regulated promoter (P-petE-vipp1) was integrated into a neutral platform in the genome of the merodiploid mutant. The copper-induced expression of P-petE-vipp1 allowed a complete segregation of the vipp1 mutant and observation of the phenotype of Synechocystis 6803 with different levels of vesicle-inducing protein in plastids 1 (Vipp1). When P-petE-vipp1 was turned off by copper deprivation, Synechocystis lost Vipp1 and photosynthetic activity almost simultaneously, and at a later stage, thylakoid membranes and cell viability. The photosystem II (PSII)-mediated electron transfer was much more rapidly reduced than the PSI-mediated electron transfer. By testing a series of concentrations, we found that P-petE-vipp1 cells grown in medium with 0.025 mu M Cu2+ showed no reduction of thylakoid membranes, but greatly reduced photosynthetic activity and viability. These results suggested that in contrast to a previous report, the loss of photosynthetic activity may not have been due to the loss of thylakoid membranes, but may have been caused more directly by the loss of Vipp1 in Synechocystis 6803.

Changes in the patterns of inorganic nitrogen and TN/TP ratio and the associated mechanism of biological regulation in the shallow lakes of the middle and lower reaches of the Yangtze River

The changes of NH3-N, NO3-N, NO2-N and TN/TP were studied during growth and non-growth season in 33 subtropical shallow lakes in the middle and lower reaches of the Yangtze River. There were significant positive correlations among all nutrient concentrations, and the correlations were better in growth season than in non-growth season. When TP > 0.1 mgL(-1), NH3-N increased sharply in non-growth season with increasing TP, and NO3-N increased in growth season but decreased in non-growth season with TP. These might be attributed to lower dissolved oxygen and low temperature in non-growth season of the hypereutrophic lakes, since nitrification is more sensitive to dissolved oxygen and temperature than anti nitrification. When 0.1 mgL(-1)> TP > 0.035 mgL(-1), TN and all kinds of inorganic nitrogen were lower in growth season than in non-growth season, and phytoplankton might be the vital regulating factor. When TP < 0.035 mgL(-1), inorganic nitrogen concentrations were relatively low and NH3-N, NO2-N had significant correlations with phytoplankton, indicating that NH3-N and NO2-N might be limiting factors to phytoplankton. In addition, TN/TP went down with decline in TIP concentration, and TN and inorganic nitrogen concentrations were obviously lower in growth season than in non-growth season, suggesting that decreasing nitrogen (especially NH3-N and NO3-N) was an important reason for the decreasing TN/TP in growth season. The ranges of TN/TP were closely related to trophic level in both growth and non-growth seasons, and it is apparent that in the eutrophic and hypertrophic state the TN/TP ratio was obviously lower in growth season than in non-growth season. The changes of the TN/TP ratio were closely correlated with trophic levels, and both declines of TN in the water column and TP release from the sediment were important factors for the decline of the TN/TP ratio in growth season.

Use of microbial community to evaluate performance of a wetland system in treating Pb/Zn mine drainage

The performance of a wetland system in treating lead (Pb)/zinc (Zn) mine drainage was evaluated by using the polyurethane foam unit (PFU) microbial community (method), which has been adopted by China as a standardized procedure for monitoring water quality. The wetland system consisted of four cells with three dominant plants: Typha latifolia, Phragmites australis and Paspalum distichum. Physicochemical characteristics [pH, EC, content of total suspended solid (TSS) and metals (Pb, Zn, Cd, and Cu)] and PFU microbial community in water samples had been investigated from seven sampling sites. The results indicated that the concentrations of Pb, Zn, Cd, Cu, and TSS in the mine drainage were gradually reduced from the inlet to the outlet of the wetland system and 99%, 98%, 75%, 83%, and 68% of these metals and TSS respectively, had been reduced in concentration after the drainage passed through the wetland system. A total of 105 protozoan species were identified, the number of protozoa species and the diversity index (DI) gradually increased, while the heterotrophic index (HI) gradually decreased from the inlet to the outlet of the wetland system. The results indicated that DI, HI, and total number species of protozoa could be used as biological indicators indicating the improvement of water quality.

Detection of extracellular phosphatases in natural spring phytoplankton of a shallow eutrophic lake (Donghu, China)

The species-specific production of extracellular phosphatases in phytoplankton of a subtropical polymictic take was investigated from March to May 2004. Phosphatase activity was detected directly at the site of enzyme action using the enzyme-labelled fluorescence (ELF) technique. Size fractionation of bulk phosphatase activity (PA), concentrations of soluble reactive phosphorus (SRP), chlorophyll a, and phytoplankton composition were determined in parallel. Phosphatase-positive cells were present in every phytoplankton sample; labelled cells were detected in 33 algal taxa, including many chlorophytes, dinoflagellates and some diatoms, but never among cyanobacteria. We recorded an unusual dinoflagellate bloom (Peridiniopsis sp.), of which similar to 25% of the cells were phosphatase-positive. Several populations were partly phosphatase-positive whenever present, while some other species never showed any activity. The production of extracellular phosphatases was not primarily regulated by ambient P concentrations; algae produced these enzymes even if SRP concentrations were high. Moreover, heterotrophic nanoflagellates most probably contributed to the pool of particle-bound PA in some samples.

Residual plastids of bleached mutants of Euglena gracilis and their effects on the expression of nucleus-encoded genes

Bleached mutants of Euglena gracilis were obtained by treatment with ofloxacin (Ofl) and streptomycin (Sm) respectively. As shown by electron microscopy, the residual plastids contain prothylakoids in an Ofl mutant, and the highly developed and tightly stacked membranous structure found in cells of two Sm, mutants. Nine genes of the plastid genome were examined with PCR, showing that ribosomal protein genes and most other plastid genes were lost in all but one Sm mutant. Using differential display and RT-PCR, it was shown that chloroplast degeneration could cause changes in transcription of certain nucleus-encoded genes during heterotrophic growth in darkness.

Changes in the plankton community following introduction of filter-feeding planktivorous fish

1. We conducted enclosure experiments in a shallow eutrophic lake, in which a biomass gradient of the filter-feeding planktivore, silver carp, Hypophthalmichthys molitrix Valenciennes, was created, and subsequent community changes in both zooplankton and phytoplankton were examined. 2. During a summer experiment, a bloom of Anabaena flos-aquae developed (approximate to 8000 cells mL(-1)) solely in an enclosure without silver carp. Concurrent with, or slightly preceding the Anabaena bloom, the number of rotifer species and their abundance increased from seven to twelve species (1700-14 400 organisms L-1) after the bloom in this fish-free enclosure. Protozoans and bacteria were generally insensitive to the gradient of silver carp biomass. 3. During an autumn experiment, on the other hand, large herbivorous crustaceans were more efficient than silver carp in suppressing the algae, partly because the lower water temperature (approximate to 24 degrees C) inhibited active feeding of this warm-water fish and also formation of algal colonies. Heterotrophic nanoflagellate and bacterial densities were also influenced negatively by the crustaceans. 4. Correspondence analysis (CA) was applied to the weekly community data of zooplankton and phytoplankton. A major effect detected in the zooplankton community was the presence/absence of silver carp rather than the biomass of silver carp, whereas that in the phytoplankton community was the fish biomass before the Anabaena bloom, but shifted to the presence/absence of the fish after the bloom.