城市河流细菌群落特征及影响因素——以鄱阳湖流域赣江南昌段为例

为了了解赣江细菌群落特征,探讨环境因子对城市河流细菌群落结构组成的影响,本文基于高通量测序分析鄱阳湖流域赣江南昌段2015年4月至2016年3月5个采样点(G1~G5)的细菌群落特征.采用R语言"Vegan"程序包中的"Bioenv"程序、冗余分析方法分析环境因子对河水细菌群落的影响,基于Bayesian统计方法的Source Tracker模型估算南昌城区对赣江细菌群落的改变比例.结果表明:赣江南昌段的优势菌门为放线菌门(Actinobacteria,34.05%)和变形菌门(Proteobacteria,31.59%),其次为厚壁菌门(Firmicutes,15.49%)和拟杆菌门(Bacteroidetes,9.12%);气温和流量是河流细菌群落主要影响因子,其中气温是影响OTU丰度的最佳单参数,影响门丰度的最佳单参数是流量;水文气象条件(气温和流量)对河流细菌群落的影响大于水化学指标,包括营养盐(溶解有机碳、总磷和铵态氮)、水化学离子(HCO_3~-、Cl~-、NO_3~-、SO_4~(2-)、Na~+、K~+和Mg~(2+))和重金属元素(Mn、Cd、Cr、Pb、Cu、As、Ba和Fe).除Proteobacteria外,其余门的细菌相对丰度在5个采样点之间不存在显著差异;Source Tracker模型结果表明南昌城区对赣江细菌群落结构组成的影响为6.33%~34.67%,对赣江南支细菌群落的影响显著高于其他河段;南昌城区对细菌群落的影响较小,主要的影响门类为Proteobacteria.     

海洋蛭弧菌DA5全基因组测序及序列分析

为深入挖掘和利用海洋蛭弧菌及其基因资源, 本研究利用Illumina HiSeq测序平台对一株属于嗜盐噬菌弧菌(Halobacteriovorax)的海洋蛭弧菌DA5进行了全基因组测序, 对基因组进行组装、基因预测和功能注释, 并与其他8株蛭弧菌进行了比较基因组分析。结果显示: DA5基因组大小为3.27Mb, GC含量为36.5%, 预测编码基因3175个。在DA5基因组中注释到303个基因具有直系同源蛋白簇分类, 与代谢通路相关基因1239个。比较基因组分析表明: DA5符合海洋蛭弧菌基因组的基本特征, 与其他8株蛭弧菌共有467个同源基因家族, 而DA5特有基因为266个。DA5中与细胞运动相关基因62个, 其中编码甲基受体趋化蛋白、鞭毛蛋白、鞭毛马达及其开关蛋白基因均与嗜盐噬菌弧菌BAL6_X的对应基因亲缘关系最近。对DA5全基因组序列的注释和功能分析, 为深入研究其捕食特性和作用机制, 并更有效地利用其防控海水养殖细菌病害提供了基础。     

病毒感染海洋球石藻Emiliania huxleyi的转录组分析

海洋球石藻Emiliania huxleyi及其特异性裂解病毒E. huxleyi virus (EhVs)在调节海洋碳、硫循环及全球气候变化中起着重要作用,也是开展真核生物病毒–宿主相互作用研究的良好模型系统之一。为了探究病毒感染条件下E. huxleyi基因表达水平的变化,以海洋球石藻E. huxley–BOF 92及其专一性裂解病毒EhV-99B1为研究对象,利用Illumina HiSeq 2000高通量测序技术,分别对E. huxleyi病毒感染组(Exp)和非感染对照组(Con)6 h和45 h的藻细胞样品进行转录组测序分析。共得到32 909条平均长度为1 153 bp的基因。病毒感染6 h和45 h分别得到2 617和5 229个差异表达基因,其中共差异表达基因465个。随机选取10条差异表达基因,采用qRT-PCR进行实验验证,结果证实转录组分析可靠。GO功能注释和KEGG通路富集,发现大量基因与氧化应激反应、脂类代谢、碳水化合物代谢及信号转导等代谢过程相关,其中变化最显著的是谷胱甘肽代谢途径。从病毒感染球石藻转录组中筛选出部分与氧化应激反应相关的基因,其中9个基因显著上调,11个基因显著下调,表明宿主能够通过体内的氧化应激反应响应病毒胁迫。     

High-throughput sequencing of 16S r RNA amplicons characterizes gut microbiota shift of juvenile sea cucumber Apostichopus japonicus feeding with three antibiotics

Sea cucumber Apostichopus japonicus is an important marine economic species in Asian countries due to its profound nutritional and medicinal value. So far, with the rapid development of intensifi ed artifi cial aquaculture of sea cucumbers, the use of antibiotics is still an inexpensive and dispensable way to treat pathogenic infections, especially during the nursery phase. However, there is little information on the eff ects of antibiotics on the intestinal microbiota of sea cucumber. Therefore an Illumina based sequencing method was used to examine the intestinal bacterial composition of juvenile A . japonicas following diets with three typical antibiotics (tetracycline, erythromycin, and norfl oxacin) under 15, 30, and 45 d. The fi ndings reveal that diff erent antibiotics have distinct eff ects on the growth performance of juvenile sea cucumbers. However, the richness and diversity of microbiota were barely aff ected by antibiotics but the community composition alterations indicated that the three antibiotics exhibited their respective patterns of reshaping the intestinal bacteria of juvenile sea cucumbers. In common, the abundance of some sensitive genera with helpful functions, such as Thalassotalea , Shewanella , Sulfi tobacter , and Halomonas decreased signifi cantly with exposure to antibiotics and the abundance of multiple potential pathogenic- and suspected antibiotic-resistant microorganisms like Arcobacter , Leucothrix , and Clostridium_sensu_stricto_1 was found increased signifi cantly in the antibiotic groups. These results suggest that low doses of antibiotics could aff ect the composition of the intestinal microbiota of sea cucumbers and might increase the risk of infection of the hosts. This study could help us to explore how antibacterial compounds modify the gut microbiota of sea cucumbers and provide theoretical guidance in hatchery management by scientifi c antibiotic use in sea cucumber mariculture.     

Effect of carbon and nitrogen ratio control on Artemia growth,water quality,biofloc microbial diversity under high salinity and zero-water exchange culture condition

Biofloc technology has been applied successfully in the intensive aquaculture of several fish and shrimp species. The growth of heterotrophic microorganisms can be stimulated through adding extra carbon, which reduces the nitrogen level in the water and provides microbial protein to the animals. However, most of the studies and practical applications have been conducted in freshwater and marine environment. This paper focused on brine shrimp Artemia that lives in high salinity environment together with other halophilic or halotolerant microorganisms. The effect of carbon supplementation on Artemia growth, water quality, and microbial diversity of biofl ocs was studied in the closed culture condition without any water exchange. The salinity of the culture medium was 100. A 24-d culture trial was conducted through supplementing sucrose at carbon/nitrogen (C/N) ratio of 5, 15, and 30 (Su5, Su15, and Su30), respectively. The culture without adding sucrose was used as a control. Artemia was fed formulated feed at a feeding ration of 60% recommended feeding level. The results showed that sucrose supplementation at higher C/N ratio (15 and 30) signifi cantly improved the Artemia survival, growth and water quality ( P <0.05). Addition of sucrose at C/N ratio of 15 and 30 significantly increased biofloc volume (BFV)( P <0.05). The Illumina MiSeq sequencing analysis showed that supplementing carbon at C/N ratio of 15 had a better total bacterial diversity and richness, and shaped the microbial composition at genera level. This study should provide information for studying the mechanism of biofloc technology and its application in high salinity culture conditions.     

长江水系不同水体鳜mtDNA控制区序列的遗传分析

对长江水系洞庭湖、鄱阳湖、秋浦河和太湖4个鳜群体共42尾的mtDNA控制区核苷酸序列进行了测定,获得了长度785 bp的同源序列.4个群体中共检测到变异位点36个,占全部序列4.6%.42个体中共检测到19种单倍型,根据碱基组成特征,19种单倍型可分为两大类型:Ⅰ型和Ⅱ型.两大类型的主要区别在第6、8、17、25、35变异位点上,Ⅰ型的核苷酸分别为G、C、G、A、C,Ⅱ型为A、T、A、G、T.除太湖群体全部表现为Ⅱ型单倍型外,不同水体鳜两大单倍型的分布频率并无明显的地理变化规律.洞庭湖、鄱阳湖、秋浦河和太湖鳜群体内的核苷酸序列差异分别为0.5%、1.0%、0.6%、0.5%,群体间的遗传差异为0.6%-0.9%.利用控制区核苷酸序列构建的NJ分子树中,各群体内的个体均未单独成群,而是互有交叉.由于遗传分化低,初步认为洞庭湖、鄱阳湖、秋浦河和太湖鳜群体可能同属一个种群--长江种群.     

阿特拉津低温降解菌的筛选及降解机理研究

以阿特拉津为唯一氮源, 在低温条件下(10℃),从吉林市污水处理厂的活性污泥中分离、筛选出1株能够高效降解地下水中阿特拉津的菌株W4.通过16S rDNA碱基测序和比对,初步确定该菌为假单胞菌属;通过室内降解条件优化,确定W4的最佳降解条件:初始pH范围为7～9,最佳碳源为蔗糖和乳糖,最佳碳源加入量为0.4 g/L.在最佳降解条件下,W4对初始质量浓度为34 mg/L、22 mg/L和10 mg/L的阿特拉津的生物降解反应符合零级反应动力学方程,对初始质量浓度为5 mg/L的阿特拉津的生物降解反应符合一级反应动力学方程.GC/MS分析结果显示,菌株W4降解阿特拉津遵循氯水解途径,代谢产物为2-羟基-4-乙胺基-6-异丙胺基-1,3,5-三嗪.     

珠江下游浮游细菌群落结构的时空分布特征

浮游细菌群落对河流变化具有高度响应性,并可能影响河流生境的生物地球化学过程.因此,了解浮游细菌群落的时空特征,阐明其在河流生态系统中的生态功能具有重要科学意义.于2016年6月和12月对珠江下游浮游细菌群落结构的时空特征进行调查研究,并采用16S rRNA高通量测序技术对样品组分进行分析.结果表明细菌群落主要由变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、抗微菌门(Verrucomicrobia)和蓝细菌门(Cyanobacteria)组成.其中变形菌门相对丰度最高,主要包括β-变形菌纲(Betaproteobacteria)、α-变形菌纲(Alphaproteobacteria)和γ-变形菌纲(Gammaproteobacteria).季节上,丰水期Shannon-Wiener和Chao 1多样性指数高于枯水期其中芽孢杆菌纲(Bacilli)和黄杆菌纲(Flavobacteriia)差异显著;空间上,珠江下游可分为西江沿线、珠三角河网中部和广州市周边3个区域.采用RDA分析表明水温(WT)、溶解氧(DO)、磷酸盐(PO_4~(3-)-P)、硅酸盐(SiO_4--Si)、总磷(TP)和透明度(SD)是驱动细菌群落变化最显著的环境因子,可影响细菌的增殖代谢.其中WT和SD是影响季节差异的重要因素将丰水期和枯水期区分开;而DO、化学需氧量(COD)和营养盐(PO_4~(3-)-P、SiO_4~(2-)-Si、TP)是影响空间差异的重要因素将西江沿线、珠三角河网中部和广州市周边区分开.PICRUSt功能预测分析表明,转运体(Transporters)、ABC转运体(ABC transporters)、DNA修复和重组蛋白(DNA repair and recombination proteins)等是珠江下游浮游细菌群落所表达的主要功能其中转运体和ABC转运体功能丰水期明显高于枯水期.研究结果可为珠江下游生态环境保护提供科学参考依据.     

健康与患病大黄鱼消化道微生物结构特征分析

为研究患病大黄鱼(Larimichthys crocea)消化道中可能的病原微生物,采用高通量测序技术详细分析了健康和患病大黄鱼的胃、幽门盲囊和肠道中微生物群的组成信息和分布特征.研究显示,变形菌门、拟杆菌门和厚壁菌门为健康和患病大黄鱼消化道的优势菌门,且其丰度在两类鱼消化道中差异显著.拟杆菌属和乳杆菌属为健康大黄鱼消...