Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii

Trachinotus blochii is one of the important commercial fish species.In this study,we aim to confirm the reliability reference genes in T.blochii during different bacterial challenge through quantitative real-time PCR(qRT-PCR).The expression of the seven selected genes in four immune organs(i.e.,spleen,kidney,intestine,and gill) stimulated with Vibrio harveyi,Edwardsiella tarda,and Streptococcus agalactiae were determined by qRT-PCR.The PCR data was analyzed using the geNorm and NormFinder algorithms.The results showed the selection of the internal controls should be tissue specific when studying gene expression in response to bacterial stimulation.After 48 h of stimulation with V.harveyi,geNorm ranked EF1 A/Actin,18 S rRNA/B2M,UBCE/B2M,and 18 S rRNA/B2M,as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.After 48 h of stimulation with E.tarda,geNorm ranked 18 S rRNA/EF1 A,18 S rRNA/B2M,B2M/RPL13,and 18 S rRNA/EF1 A,as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.After 48 h of stimulation with S.agalactiae,18 S rRNA/EF1 A,18 S rRNA/B2 M,B2 M/Actin,and 18 S rRNA/B2M were ranked as the most stably expressed genes in spleen,kidney,intestine,and gill,respectively.Compared to the results analyzed by geNorm,reference genes received similar rankings when using NormFinder software.The results showed that the reference genes appeared to be not only tissue specific,but also specific to the infecting species of bacteria.If one gene is preferred when T.blochii were infected by bacteria,18 S rRNA,B2M,B2M,18 S rRNA may be used in spleen,kidney,intestine,and gill,respectively.     

Pyrosequencing of nirS gene revealed spatial variation of denitrifying bacterial assemblages in response to wetland desertification at Tibet plateau

Amplicon sequencing of functional genes is a powerful technique to explore the diversity and abundance of microbes involved in biogeochemical processes. One such key process, denitrification, is of particular importance because it can transform nitrate(NO3-) to N2 gas that is released to the atmosphere. In nitrogen limited alpine wetlands, assessing bacterial denitrification under the stress of wetland desertification is fundamental to understand nutrients, especially nitrogen cycling in alpine wetlands, and thus imperative for the maintenance of healthy alpine wetland ecosystems. We applied amplicon sequencing of the nirS gene to analyze the response of denitrifying bacterial community to alpine wetland desertification in Zoige, China. Raw reads were processed for quality, translated with frameshift correction, and a total of 95,316 nirS gene sequences were used for rarefaction analysis, and 1011 OTUs were detected and used in downstream analysis. Compared to the pristine swamp soil, edaphic parameters including water content, organic carbon, total nitrogen, total phosphorous, available nitrogen, available phosphorous and potential denitrification rate were significantly decreased in the moderately degraded meadow soil and in severely degraded sandy soil. Diversity of the soil nirS-type denitrifying bacteria communities increased along the Zoige wetland desertification, and Proteobacteria and Chloroflexi were the dominant denitrifying bacterial species. Genus Cupriavidus(formerly Wautersia), Azoarcus, Azospira, Thiothrix, and Rhizobiales were significantly(P0.05) depleted along the wetland desertification succession. Soil available phosphorous was the key determinant of the composition of the nirS gene containing denitrifying bacterial communities. The proportion of depleted taxa increased along the desertification of the Zoige wetland, suggesting that wetland desertification created specific physicochemical conditions that decreased the microhabitats for bacterial denitrifiers and the denitrification related genetic diversity.     

Bacterial community shifts in different functional zones of a reservoir with ecological purification facilities

Ecological purification in a reservoir is an important strategy to control the level of nutrients in water.The bacterial community of such a reservoir is the main agent for pollutant degradation,which has not been fully documented.Taking the Jinze Reservoir,a freshwater source for Shanghai,China as the case,its spatial distributions of water and sediment bacteria were determined using 16 S rRNA gene-based Illumina MiSeq sequencing,and the environmental parameters were analyzed.The reservoir takes natural river water and functions as an ecological purification system,consisting of three functional zones,i.e.,pretreatment zone,ecological purification zone,and ecological sustaining zone.Results show that the concentrations of both total nitrogen(TN) and total phosphorus(TP) decreased considerably after the ecological treatment,and the concentration of dissolved oxygen(DO) in the ecological purification zone was boosted from that before pretreatment.In addition,patterns of bacterial communities in both water and sediment were similar and consisted of mainly Proteobacteria,Actinobacteria,and Bacteroidetes.However,difference in water bacterial composition was distinct in each functional zone,whereas the bacterial communities in sediment changed only slightly among sediment samples.Network analysis revealed nonrandom co-occurrence patterns of bacterial community composition in water and sediment,and Pseudomonas,unclassified Comamonadaceae,Variovorax,and Dechloromonas were the key taxa in the co-occurrence network.The bacterial taxa from the same module of the network had strong ecological connections,participated in C-cycles,and shared common trophic properties.PICRUSt analysis showed that bacteria were involved potentially in various essential processes;and the abundance of predicted xenobiotic biodegradation genes showed a decreasing trend in water samples from the inlet to the outlet of the reservoir.These results improve our current knowledge of the spatial distribution of bacteria in water and sediment in ecological purification reservoirs.     

Eukaryotic microbial distribution pattern and its potential effects on fisheries in the fish reserves of Qiantang River in breeding season

To examine the eukaryotic biodiversity of aquatic ecosystems in the Qiantang River,China,eukaryotic microbes in the river were investigated using 18 S rRNA gene sequencing during the breeding season(July to August 2018).Four distinct distribution patterns(1.Jiande;2.Tonglu and Fuyang;3.Jiubao;4.Yanguan) of the microbial community and their potential effects on fishery activities were observed.Results show lower abundances of Dinophyta and Fungi and higher abundances of Cryptophyta and Chlorophyta in Tonglu and Fuyang than those in the other three sections.In addition,the reserves(Tonglu and Fuyang)destabilized the original eukaryotic microbial co-occurrence network.Among all the environmental factors measured,nitrogen(nitrite,nitrate,ammonium),water temperature and total chlorophyll a acted as major driving factors that controlled the eukaryotic microbial distribution.Furthermore,the existence of some algae(e.g.,Chrysophyceae,Cryptophytes,and Chlorophyceae) and fungi(e.g.,Rhizophydium) in Tonglu and Fuyang was beneficial to juvenile fish growth and water quality,although some detrimental species(e.g.,Aphanomyces) needed attention.This study provides further insights into the sustainable protection and utilization of rivers.     

基于室内路网的跨楼层路径规划技术的设计与实现

随着室内定位技术、BIM-GIS技术和室内地图研究的发展,室内位置服务需求日益旺盛。作为室内位置服务的关键技术之一,室内路径规划成为研究与应用热点。但是早先的路径规划技术往往基于单层网络拓扑模型而设计,由于模型缺少楼层之间的连通信息,而不能实现跨楼层导航算法分析;而后随着跨楼层导航算法的出现,跨楼层的路径规划、导航及定位得以实现,但这些跨楼层导航算法基于楼层连接通达规则,往往需要多次遍历楼层信息,运算效率低下。针对这一问题,本文利用电梯、扶梯、楼梯等楼层间连接信息,设计、实现了基于楼层连接表的室内多层路网分析模型,并采用逐层分析的方法和多次网络分析,实现室内跨楼层的带有多个途经点路径分析。整个技术框架包括跨楼层路网生成模块、位置点接收模块、网络分析模块、循环控制模块。算法方面,本研究采用双向遍历的A*最短路径分析算法,通过网络路径分析,最终将满足约束条件的规划路径显示在移动设备上,帮助用户实现跨楼层、多途经点的定位与路径导航。     

Effects of Silver Nanoparticles on Denitrification and Associated N2O Release in Estuarine and Marine Sediments

Silver nanoparticles(AgNPs)have been widely used in medicine and consumer products.And it enters the river in dif-ferent ways,then finally converges to the ocean through the estuary.AgNPs polution can affect NO₂?and N₂O production by denitri-fiers in aquatic system.The effects of AgNPs on denitrification activity,nitrogen transformation and nitrous oxide(N₂O)emission were investigated in Dagu River Estuary(DRE)and Jiaozhou Bay(JZB).The results showed that the potential denitrification activity(PDA),NO₃?and NO₂?reduction rates decreased with an increase of AgNPs concentration in DRE and JZB.However,the N₂O ac-cumulation was significantly increased at AgNPs concentrations above 5 mg kg^?1in both areas,and the accumulation rate was greater in estuary than in bay(P<0.05).Moreover,the total bacterial count showed a slightly increasing trend with an increase of AgNPs concentration(P>0.05)in DRE and JZB.Importantly,the relative abundance of narG,nirS and nosZ gene in two areas decreased with the increase of AgNPs concentration,and the negative effect of AgNPs varied in order:nosZ>nirS>narG,inferring that the expression of denitrifying related genes could be significantly and differently inhibited by AgNPs addition.Thus,this study demon-strated that the inhibitory effect of AgNPs on different denitrification process,which may lead to the increase of inorganic nitrogen accumulation and N₂O realease.This study provides a scientific basis for the further studies of AgNPs on the ecological impact mechanism and environmental effects of offshore sedimentary environment.     

土地利用规划信息的社会化与网络化应用

针对土地利用规划信息量大增,传统管理方式无法满足新形势要求的现状,探索出一种运用GIS、网络、数据库等关键性核心技术,实现土地利用规划信息社会化与网络化的设计方案,并在GIS平台上开发了土地利用规划管理系统。该系统集图形和属性数据为一体,包括B/S与C/S两子系统,实现规划辅助编制、规划成果管理、专题图提取及规划信息发布等功能。实践证明:该方案对于改变传统管理方式,提高政府的管理与服务水平和宏观调控能力。     

Characterization and in-vivo evaluation of potential probiotics of the bacterial flora within the water column of a healthy shrimp larviculture system

A thorough understanding of the normal bacterial flora associated with shrimp larviculture systems contributes to probiotic screening and disease control.The bacterial community of the water column over a commercial Litopenaeus vannamei larval rearing run was characterized with both culture-dependent and culture-independent methods.A total of 27 phylotypes at the species level were isolated and identified based on 16 S rDNA sequence analysis.Denaturing gradient gel electrophoresis(DGGE) analysis of the V3–V5 region of 16 S rRNA genes showed a dynamic bacterial community with major changes occurred from stages zoea to mysis during the rearing run.The sequences retrieved were affiliated to four phyla,Proteobacteria,Actinobacteria,Bacteroidetes,and Firmicutes,with the family Rhodobacteraceae being the most frequently recovered one.Subsequently,13 representative strains conferred higher larval survival than the control when evaluated in the in-vivo experiments;in particular,three candidates,assigned to Phaeobacter sp.,Arthrobacter sp.,and Microbacterium sp.,significantly improved larval survival( P 0.05).Therefore,the healthy shrimp larviculture system harbored a diverse and favorable bacterial flora,which contribute to larval development and are of great importance in exploiting novel probiotics.     

Impact of Phenanthrene on Denitrification Activity and Transcription of Related Functional Genes in Estuarine and Marine Sediments

The effects of phenanthrene(Phe)on the denitrification activity and denitrifying genes(narG,nirS and nosZ)were evaluated by dose-response experiments in sediments of Dagu River Estuary(DRE)and Jiaozhou Bay(JZB).The results showed that potential denitrification activity(PDA),N2O,NO3−and NO2−reduction rates of both areas were inhibited with an increase of Phe concentrations.The PDA,N2O,NO3−and NO2−reduction rates of both areas was highest and lowest in the control(DRE:0.453,0.427,7.439 and 3.222mgNkg−1 h−1,JZB:0.592,0.555,8.470 and 3.793mgNkg−1 h−1)and highest Phe amended treatments(DRE:0.069,0.001,4.486,and 1.563 mgNkg−1 h−1;JZB:0.114,0.024,5.527 and 2.200 mgNkg−1 h−1).The inhibition rate of PDA was highest,follow by NO2−reduction and then NO3−reduction.Moreover,with the increasing of Phe concentrations,total bacteria count and the abundance of denitrifying genes were decreased.And N2O accumulation was promoted with the addition of Phe for both areas.Based on the comparison of EC50 values,denitrifiers harboring three genes were more sensitive to Phe than PDA,and denitrifiers harboring nirS gene were more sensitive,followed by nosZ gene,and then narG gene.Furthermore,according to correlation analysis,the relative abundance of denitrifying genes was much more positively correlated with PDA,NO3−and NO2−reduction than total bacteria count.In addition,the denitrification activity and total bacteria count in JZB were more inhibited than that of DRE.This study is useful for understanding the impact of Phe pollution on denitrification in estuary and marine sediments,with profound implications for the management of aquatic ecosystems regarding eutrophication(N-removal)and greenhouse effect.     

Antifouling effect of bioactive compounds from selected marine organisms in the Obhur Creek,Red Sea

Three species of sponges and a tunicate were collected from Obhur creek of Jeddah coast for this bioactivity study. In order to assess the antifouling efficacy of selected marine organisms, methanolic extracts of these organisms were tested against different fouling bacterial forms and II-instar stage of the barnacle, Balanus amphitrite. Antibiosis, bioactivity and followed by multivariate analyses were carried out to check the efficacy of antifouling effect of the selected marine organisms. Principal component analysis revealed the exemplary antifouling efficacy of the sponge extracts of Stylissa sp. observed followed by Hyrtios sp. against bacterial forms in the laboratory study. De-trended correspondence analysis confirmed that the contribution of antifouling efficacy of the selected sponge extracts was observed to be more towards Bacillus sp., Vibrio sp. and Alteromonas sp. Moreover, the efficacy of Hyrtios sp. extract(20.430 μg m L~(-1)) followed by Stylissa sp.(30.945 μg m L~(-1)) showed higher against barnacle instar compared with other extracts in the bioactivity assay. Bray-Curtis cluster analysis under paired linkage categorized all the sponge extracts into one major cluster with 75% similarity, and one outlier tunicate. More than 80% similarity observed between Hyrtios sp. and Stylissa sp. Fourier transform infrared spectroscopy(FTIR) showed that the contribution of major peaks found in the marine organisms were towards sulfones, sulfoxides, cyanates and ketones.     

Antifouling activities of marine bacteria associated with sponge (Sigmadocia sp.)

The present study aimed at assessing the antifouling activity of bacteria associated with marine sponges. A total of eight bacterial strains were isolated from the surface of sponge Sigmadocia sp., of them, SS02, SS05 and SS06 showed inhibitory activity against biofilm-forming bacteria. The extracts of these 3 strains considerably affected the extracellular polymeric substance producing ability and adhesion of biofilm-forming bacterial strains. In addition to disc diffusion assay, microalgal settlement assay was carried out with the extracts mixed with polyurethane wood polish and coated onto stainless steel coupons. The extract of strain SS05 showed strong microalgal settlement inhibitory activity. Strain SS05 was identified as Bacillus cereus based on its 16S rRNA gene. Metabolites of the bacterial strains associated with marine invertebrates promise to be developed into environment-friendly antifouling agents.     

Antimicrobial activities of novel cultivable bacteria isolated from marine sponge Tedania anhelans

Marine sponge Tedania anhelans distributes throughout the intertidal zone of Fujian, southeastern China, and is a potential source of natural bioactive products. The sponge harbors a large number of bacterial groups that have been identified using various techniques, including fluorescent in situ hybridization (FISH). Fractionation of dissociated sponge allowed isolation of 25 bacterial species. Based on 16S rRNA gene sequencing, phylogenetic analysis attributed most of these eubacteria to α-Proteobacteria, γ-Proteobacteria, Cytophaga/Flavobacterium/Bacteroidetes (CFB group), and the family Bacillaceae of Gram-positive bacteria. In sequence similarity, five putatively novel species were identified with less than 98% similarity to other strains in the NCBI database. Tests for antimicrobial activities were performed against Gram-positive bacteria, Gram-negative bacteria, fungi, antitumor indicators Escherichia coli 343/591 (with DNA repair deficiency), regular E. coli 343/636 (with different DNA repair capacity), and 10 bacterial isolates exhibited inhibitory bioactivities. Among these strains, three isolates were detected involving function gene NRPS-A domains, which were most closely related to the amino acid sequences of linear gramicidin synthetase and pyoverdine synthetase. These results contribute to our knowledge of the microbes associated with marine sponges and further reveal novel bacterial resources for the screening of bioactive marine natural products.     

老挝北部Sentinel-2 A/B云覆盖适宜阈值界定及其影像获取概率时空差异

光学卫星影像云覆盖时空特征评估是衡量其作为重要遥感监测数据源的前提。Sentinel-2 A/B影像因其免费获取、多光谱（红边）、更高时空分辨率等优势,已在全球不同尺度陆面植被与生态监测中受到重视。相较于Landsat等同类影像产品,有关Sentinel-2 A/B的云覆盖分析还未见报道。本文利用2016—2018年老挝北部所有5288景Sentinel-2 A/B影像（Granule/Tile）的云覆盖元数据,基于不同云覆盖阈值（0~100%）水平下的影像获取概率差异确定了影像获取概率分析的云覆盖适宜阈值,并揭示了云量特征阈值水平下的影像获取概率时空差异。主要结论如下:① Sentinel-2 A/B影像获取概率分析云覆盖适宜特征阈值为20%（即云覆盖≤20%）,该阈值水平下老挝北部Sentinel-2 A/B影像的逐月累积平均获取概率最高（约27.41%）;② 在20%云覆盖阈值水平下,老挝北部Sentinel-2 A/B影像逐月累积平均获取概率差异在时间上与旱季（11月—次年4月）雨季（5月—10月）的时间分布较为吻合。旱季获取概率约为42.91%,3月概率（50.27%）最大,4月与2月次之,时间上与刀耕火种焚烧与橡胶林落叶特征吻合;雨季相应概率约为11.81%,6月最低（约1.26%）;③ 老挝北部Sentinel-2 A/B影像逐月累积平均获取概率在空间上存在东西差异,旱季西部省域单元（如琅南塔）影像获取概率远高于东部,雨季西部地区影像获取概率则略低于东部地区。本研究既可为后续开展大区及全球Sentinel-2 A/B影像云量分析提供借鉴,也对开展联合国减少森林砍伐和退化排放（UN-REDD）计划引发的土地利用变化如刀耕火种农业演变、橡胶林扩张等遥感监测有指导意义。     

鄂尔多斯盆地多级次地下水流系统中硝酸盐分布特征及其成因

水资源短缺的鄂尔多斯盆地内地下水遭受硝酸盐（NO₃-）污染等问题日益突出，识别盆地不同地下水流系统的NO₃-分布规律及其成因，对地下水资源的合理利用与保护具有重要意义。选取鄂尔多斯盆地北部湖泊集中区白垩系地下水系统为研究对象，基于水化学和聚类-主成分分析划分地下水流系统级次，在此基础上对比分析不同级次地下水流系统中NO₃-分布特征，综合水化学和环境同位素分析识别多级次地下水流系统中NO₃-来源及其潜在过程。研究表明:研究区ρ（NO₃-）超出地下水质量标准（GB/T 14848-2017）Ⅲ类水标准的地下水样品集中在局部-中间地下水流系统，其超标率达到28%；区域地下水流系统中ρ（NO₃-）均值约为1 mg/L。研究区不同级次地下水流系统中ρ（NO₃-）分布特征主要与人类活动影响程度有关，而地下水蒸发富集和反硝化衰减作用对ρ（NO₃-）的影响可以忽略。其中，局部-中间地下水流系统受到人类活动产生的污染影响显著，其NO₃-污染主要来源于无机铵肥和粪便污水等；区域地下水流系统可能尚未受到人类活动污染，其NO₃-来源于天然有机氮矿化。      

Metabolic and phylogenetic profile of bacterial community in Guishan coastal water (Pearl River Estuary), South China Sea

Characteristics of a microbial community are important as they indicate the status of aquatic ecosystems. In the present study, the metabolic and phylogenetic profile of the bacterioplankton community in Guishan coastal water(Pearl River Estuary), South China Sea, at 12 sites(S1–S12) were explored by community-level physiological profiling(CLPP) with BIOLOG Eco-plate and denaturing gradient gel electrophoresis(DGGE). Our results showed that the core mariculture area(S6, S7 and S8) and the sites associating with human activity and sewage discharge(S11 and S12) had higher microbial metabolic capability and bacterial community diversity than others(S1–5, S9–10). Especially, the diversity index of S11 and S12 calculated from both CLPP and DGGE data(H 3.2) was higher than that of others as sewage discharge may increase water nitrogen and phosphorus nutrient. The bacterial community structure of S6, S8, S11 and S12 was greatly influenced by total phosphorous, salinity and total nitrogen. Based on DGGE fingerprinting, proteobacteria, especially γ- and α-proteobacteria, were found dominant at all sites. In conclusion, the aquaculture area and wharf had high microbial metabolic capability. The structure and composition of bacterial community were closely related to the level of phosphorus, salinity and nitrogen.     

Comparison of nonlinear and linear PCA on surface wind, surface height, and SST in the South China Sea

We compared nonlinear principal component analysis (NLPCA) with linear principal component analysis (LPCA) with the data of sea surface wind anomalies (SWA), surface height anomalies (SSHA), and sea surface temperature anomalies (SSTA), taken in the South China Sea (SCS) between 1993 and 2003. The SCS monthly data for SWA, SSHA and SSTA (i.e., the anomalies with climatological seasonal cycle removed) were pre-filtered by LPCA, with only three leading modes retained. The first three modes of SWA, SSHA, and SSTA of LPCA explained 86%, 71%, and 94% of the total variance in the original data, respectively. Thus, the three associated time coefficient functions (TCFs) were used as the input data for NLPCA network. The NLPCA was made based on feed-forward neural network models. Compared with classical linear PCA, the first NLPCA mode could explain more variance than linear PCA for the above data. The nonlinearity of SWA and SSHA were stronger in most areas of the SCS. The first mode of the NLPCA on the SWA and SSHA accounted for 67.26% of the variance versus 54.7%, and 60.24% versus 50.43%, respectively for the first LPCA mode. Conversely, the nonlinear SSTA, localized in the northern SCS and southern continental shelf region, resulted in little improvement in the explanation of the variance for the first NLPCA.     

Production of Pacific White Shrimp Polycultured with Swimming Crab at Different Densities,and Nutrient Budget in the Enclosure System

This study was to evaluate the effect of stocking densities on the nitrogen and phosphorus budgets for a polyculture of the crab Portunus trituberculatus and the Pacific white shrimp Litopenaeus vannamei.The shrimps(initial weight,0.012 g)were cultured at a density of 45 shrimp m^-2,and juvenile crabs(initial weight,0.024 g)were cultured at five densities of 0,3,6,9,and 12 crabs m^-2.The treatments were grouped as C0S45,C3S45,C6S45,C9S45,and C12S45,respectively.Water quality parameters,growth of shrimp and crabs,and nitrogen and phosphorus budget were measured.The results indicated that the shrimp performances in polyculture treatments C3S45 and C6S45 were superior to those in the crab-free treatment(C0S45).The crab in treatments C3S45 and C6S45 exhibited a significantly higher final mean weight and carapace width/length than those in treatments C9S45 and C12S45.The final size and survival of crabs had a negative correlation with the increasing crab stocking density.The contents of total phos-phorus and total nitrogen and the comprehensive contamination index values were higher in the C9S45 and C12S45 treatments than in the other treatments.The conversion ratios of nitrogen for crab and shrimp growth in treatment C3S45 were significantly higher than those in the crab-free treatment.These findings indicate that polyculturing shrimp with crabs at suitable densities can improve productivity,profitability,nutrient utilization,and the environmental quality.From the 60-day treatments,the optimal culture densi-ties were 3-6 crabs m^-2 and 45 shrimps m^-2.     

Life history, secondary production and trophic basis of two dominant mayflies in a subtropical stream of China

Mayflies constitute a major part of macroinvertebrate biomass and production in Iotic ecosystems, and play an important role in material cycle and energy flow. There are more than 250 species of mayflies in rivers and streams of China. In order to learn their ecological functions, an investigation on life cycle, production and trophic basis of dominant species of mayflies in a second-order branch of Hanjiang River basin, Hubei, China was carried out during June 2003 to June 2004. The results showed that the dominant mayfly species Epeorus sp. and Caenis sp. developed two generations per year; in term of Epeorus sp., pupation mainly occurred in spring and then from late summer to early autumn, while Caenis sp. pupated in spring and autumn. The abundance and biomass of the Epeorus sp. population peaked twice （1 226 ind/m^2, 3.142 5g/m^2） in April and June. Caenis sp. also had two peaks （307ind/m^2, 1.590 g/m^2）, but in February and June. Cohort production and cohort P/B ratio of Epeorus sp. were 161.009 g/m2 wet weight and 7.7, respectively, and annual production and P/B ratio were 267.46g/m^2.a wet weight and 15.4, respectively; cohort production and P/B ratio of Caenis sp. were 26.7995g/m^2 wet weight and 4.7, its annual production and P/B ratio were 53.60 g/m2.a wet weight and 9.4, respectively. For Epeorus sp., the proportions contributing to secondary production of the main food types were： amorphous detritus, 33.46%; fungi, 10.83%; vascular plant detritus, 1.80%; diatoms, 53.90%; for Caenis sp., the proportions were 70.79%, 6.90%, 3.52% and 18.77%, respectively.