洄游型、淡水型和陆封型刀鲚的寄生蠕虫群落结构

由于对淡水环境的适应和江湖阻隔,部分洄游型刀鲚(Coilia nasus)分化成淡水型和陆封型,为寻找区分洄游型、淡水型和陆封型这3种生态型刀鲚的寄生虫标志,并分析其寄生蠕虫群落结构特征,本文调查了3种生态型刀鲚鳃部、胃、肠和幽门盲囊中寄生蠕虫的种类和感染情况。共发现10种寄生虫,包括鳃上3种单殖吸虫:林氏异钩铗虫(Heteromazocraes lingmueni)、细长嗜鳀虫(Helciferus tenuis)和长江中华钩铗虫(Sinomazocraes changjiangensis);胃部1种复殖吸虫:鲚套茎吸虫(Elytrophallus coiliae);肠道3种线虫:简单异尖线虫(Anisakis simplex)、对盲囊线虫(Contracaecum sp.)和胃瘤线虫(Eustrongylides sp.),以及肠和幽门盲囊的3种棘头虫:陈氏刺棘虫(Acanthosentis cheni)、鲇异吻钩棘头虫(Arhythmacanthus parasiluri)、长江傲刺棘头虫(Brentisentis yangtzensis)。林氏异钩铗虫和陈氏刺棘虫在3种生态型的刀鲚中都有较高的感染率和平均丰度;海水性寄生虫细长嗜鳀虫、鲚套茎吸虫和对盲囊线虫只感染洄游型刀鲚,可作为区分洄游型刀鲚的寄生虫标志;淡水性寄生虫长江中华钩铗虫、鲇异吻钩棘头虫和长江傲刺棘头虫只在淡水型刀鲚中发现,可作为淡水型刀鲚的寄生虫标志。洄游型刀鲚的物种丰富度和Brillouin多样性最高,分别在1.25和0.19以上,淡水型刀鲚的较低,分别为0.79~1.12和0.10~0.12,陆封型刀鲚的最低,分别在0.66和0.02以下,这主要是由于淡水型和陆封型刀鲚中海水性寄生虫大部分丢失,以及淡水性寄生虫感染率和平均丰度都较低造成的。盐度降低和中间宿主缺乏是导致淡水型和陆封型刀鲚中海水性寄生虫丢失的主要原因;而淡水性寄生虫较低的感染水平可能是由于其对新宿主的适应时间较短;地理隔离可能是陆封型刀鲚中寄生虫群落多样性最低的另一重要原因。     

光唇鱼仔稚幼鱼肠道菌群与养殖水体细菌群落的相关性

采用16S rRNA高通量测序,系统研究了光唇鱼(Acrossocheilus fasciatus)仔鱼、稚鱼和幼鱼肠道菌群组成及与同时期养殖水体细菌群落的相关性。研究结果表明,光唇鱼仔鱼、稚鱼和幼鱼的肠道菌群中Chao1指数和Shannon指数均没有显著变化(P>0.05);而随着光唇鱼幼体的发育,养殖水体中Chao1指数和Shannon指数呈现显著下降趋势(P<0.05)。光唇鱼仔鱼和稚鱼肠道菌群中的优势菌门由变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)组成,而同时期养殖水体中优势菌门为变形菌门;光唇鱼幼鱼肠道菌群中的优势菌门为梭杆菌门(Fusobacteria)和变形菌门,同时期养殖水体中优势菌门由变形菌门、拟杆菌门和梭杆菌门组成。线性回归分析结果显示,随光唇鱼幼体发育,在光唇鱼肠道菌群中变形菌门、厚壁菌门和梭杆菌门相对丰度的时序变化趋势与其在养殖水体中相同。在属水平上,光唇鱼仔鱼、稚鱼肠道菌群中优势菌属均为醋酸杆菌属(Acetobacter),而幼鱼肠道菌群中优势菌属为鲸杆菌属(Cetobacte...     

三种室内饲养鱼类肠道微生物群落PCR-DGGE指纹分析

以室内饲养的斑点叉尾(Ictalurus punctatus)、银鲫和异育银鲫(中科三号)(Carassius auratus gibelio)幼鱼为对象,通过PCR-DGGE指纹技术对其肠道微生物群落进行了探索研究。在三种鱼的肠道中检测到不同的PCR-DGGE指纹谱带,其中斑点叉尾的平均谱带数(7.5)相对于银鲫和异育银鲫的谱带数(分别为15和14)要少。基于PCR-DGGE指纹谱带及各谱带相对丰度的UPGMA聚类和MDS排序结果显示银鲫和异育银鲫肠道微生物相似性高,而与斑点叉尾的差异比较大;rank-abundance散点图及回归分析也显示斑点叉尾与银鲫、异育银鲫肠道微生物群落存在显著差异(P0.05),而银鲫和异育银鲫之间无显著差异(P=0.383)。在斑点叉尾肠道中检测到的菌群主要是变形杆菌,包括-变形杆菌和-变形杆菌;而银鲫和异育银鲫肠道中菌群主要包括梭杆菌属中的类群,还包括变形杆菌门中的气单胞菌属,以及一些未知的类群。以上结果均表明在所研究的三种鱼的幼鱼阶段,其肠道微生物组成在不同种类鱼中存在差异,且该差异受基因型的影响可能更大。       

综合养殖池塘中三角帆蚌和鱼类肠道细菌的组成

采用 PCR-DGGE (PCR-denaturing gradient gel electrophoresis)方法研究了综合养殖池塘中三角帆蚌(Hyriopsis cumingii)、草鱼(Ctenopharyngodon idella)、鳊(Parabramis pekinensis)、银鲫(Carassius auratus gibelio)、青鱼(Mylopharyngodon piceus)和鳙(Aristichthys nobilis)的肠道细菌组成,并分析了水体中的浮游细菌对鱼、蚌肠道细菌的影响。研究结果表明,三角帆蚌和混养鱼类肠道细菌属于厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、疣微菌门(Verrucomicrobia)、蓝细菌(Cyanobacteria)、梭杆菌门(Fusobacteria)。其中,三角帆蚌肠道优势菌群为不动杆菌属(Acinetobacter)、志贺氏菌属(Shigella)和分枝杆菌属(Mycobacterium),草鱼肠道优势菌群为梭菌属(Clostridium),鳊肠道优势菌群为梭菌属和假单胞菌属(Pseudomonas),银鲫肠道优势菌群为不动杆菌属和志贺氏菌属,青鱼肠道优势菌群为梭菌属和不动杆菌属,鳙肠道优势菌群为不动杆菌属和弧菌属(Vibrio)。三角帆蚌与银鲫肠道细菌谱带相似性较高,草鱼与鳊肠道细菌谱带相似性较高,表明鱼类肠道细菌组成特点与食性存在一定的关系。水体浮游细菌优势菌群为类芽孢杆菌属(Paenibacillus)。三角帆蚌及鱼类肠道细菌群落与水体浮游细菌群落组成相似性较低,表明水体浮游细菌对三角帆蚌和鱼类肠道细菌优势菌群的影响有限。     

高尿酸血症鹌鹑肠道菌群结构分析

目的探讨高尿酸血症鹌鹑肠道菌群结构变化特点,为高尿酸血症病理机制研究提供参考。方法将迪法克鹌鹑随机分为两组:正常组和模型组,正常组喂饲正常饲料,模型组喂饲高嘌呤饲料; PCR-DGGE法构建两组鹌鹑盲肠内容物菌群DNA指纹图谱,对图谱进行数字化处理,聚类分析法比较两组鹌鹑肠道菌群结构整体差异,logistic回归法辨别差异条带,克隆和鉴定差异条带序列; HE染色观察盲肠组织结构变化。结果造模第28天,模型组鹌鹑与正常组比较,血尿酸显著升高、肠道菌群整体结构发生变化,DGGE指纹图谱上12条差异条带,序列鉴定归属于拟杆菌门、厚壁菌门、变形菌门;盲肠组织出现炎性变化。结论高嘌呤饮食诱导的高尿酸血症鹌鹑肠道菌群结构发生改变。     

喂养不同饵料对匙吻鲟和鳙消化道微生物区系的影响

为了解消化道的菌群结构及影响因素, 用PCR-DGGE技术和序列分析调查投喂鲜活饵料(BI-L)和配合饲料(BI-C)的鳙肠道菌群, 和投喂鲜活饵料(PS-L, PI-L)和配合饲料(PS-C, PI-C)的匙吻鲟胃和肠道菌群。实验共回收测序16条DGGE条带, 系统发育分析显示鳙和匙吻鲟所有待测样本的优势菌为厚壁菌门(31.25%, 5条)。其他属于-变形菌纲(-Proteobacteria)(25%, 4条)、拟杆菌门(Bacteroidetes)(12.5%, 2条)、梭杆菌门(Fusobacteria)(12.5%, 2条)、蓝藻门(Cyanobacteria)(6.25%, 1条)、放线菌门(Actinobacteria)(6.25%, 1条)、-变形菌门(-Proteobacteria)(6.25%, 1条)。不同饵料喂养的匙吻鲟肠道菌群具有较高的相似性(49%), 不同饵料喂养的鳙肠道菌群也具有较高的相似性(59%), 说明这些生态位中存在着更稳定成型的微生物群落。然而, 两组不同饵料喂养的匙吻鲟胃样本菌群之间存在相当大的差异, 表明匙吻鲟胃部菌群可能更易受到饵料的影响。     

喂养不同饵料对匙吻鲟和鳙消化道微生物区系的影响（英文）

为了解消化道的菌群结构及影响因素,用PCR-DGGE技术和序列分析调查投喂鲜活饵料(BI-L)和配合饲料(BI-C)的鳙肠道菌群,和投喂鲜活饵料(PS-L,PI-L)和配合饲料(PS-C,PI-C)的匙吻鲟胃和肠道菌群。实验共回收测序16条DGGE条带,系统发育分析显示鳙和匙吻鲟所有待测样本的优势菌为厚壁菌门(31.25%,5条)。其他属于γ-变形菌纲(γ-Proteobacteria)(25%,4条)、拟杆菌门(Bacteroidetes)(12.5%,2条)、梭杆菌门(Fusobacteria)(12.5%,2条)、蓝藻门(Cyanobacteria)(6.25%,1条)、放线菌门(Actinobacteria)(6.25%,1条)、β-变形菌门(β-Proteobacteria)(6.25%,1条)。不同饵料喂养的匙吻鲟肠道菌群具有较高的相似性(49%),不同饵料喂养的鳙肠道菌群也具有较高的相似性(59%),说明这些生态位中存在着更稳定成型的微生物群落。然而,两组不同饵料喂养的匙吻鲟胃样本菌群之间存在相当大的差异,表明匙吻鲟胃部菌群可能更易受到饵料的影响。     

饵料对鳡肠道微生物多样性的影响

通过PCR-DGGE指纹分析并结合克隆、测序对饲喂人工配合饲料和冰鲜鱼两种不同饵料的鳡肠道微生物群落结构及多样性差异进行了比较研究。摄食配合饲料和冰鲜鱼的鳡肠道样品中分别检测到21条和17条清晰的DGGE指纹条带; 进一步的克隆、测序及BLAST比对分析表明, 21条测序谱带与GenBank数据库中已知微生物的同源性为98%100%。配合饲料饲养鳡肠道微生物特有条带代表种群主要为魏斯氏菌(Weissellakoreensis)等, 冰鲜鱼饲养鳡特有条带代表种为威斯康星米勒菌(Moellerella wisconsensis)等。从PCR-DGGE指纹相似性来看, 不同饵料饲养鳡的肠道细菌组成差异较为明显, 相似性仅为11.9%42.6%。鳡肠道菌群的DGGE 指纹图谱中条带的H'指数(Shannon-Weiner 指数)最高为配合饲料饲养鳡第Ⅴ组样本, 达到2.84, 最低的为冰鲜鱼饲喂下的鳡第Ⅵ组样本, 为2.46。研究结果表明, 投喂人工配合饲料和冰鲜鱼会对鳡肠道菌落产生影响, 可为鳡饲料的开发提供一定的基础依据。此外, 两类鳡的肠道群落PCR-DGGE指纹图谱有助于这两种鳡产品的跟踪和肠道益生菌研究。     

刀鲚幼鱼消化系统的组织形态学结构

采用光镜和扫描电镜观察长江刀鲚(Coilia nasus)幼鱼消化系统组织形态学结构。结果显示,刀鲚体长,口裂大,含有犬齿状的颌齿和尖锥状的腭齿,具有5对鳃弓,鳃耙长度明显大于鳃丝且表面附着不规则绒毛状细齿;胃呈"Y"型,胃与肠连接处具有16～21个指状环形幽门盲囊;肠为直肠,较短,比肠长为0.241±0.080;肝分为两叶,胰为独立的器官。刀鲚口咽腔为复层鳞状上皮,含有腺体、大量椭圆形黏液细胞、少量杯状细胞及味蕾;胃黏膜都为典型的单层柱状上皮,含有较多由上皮凹陷形成的胃小凹和胃腺;幽门盲囊具有20～25个丰富的褶皱,占满大部分幽门盲囊腔,黏膜层具有微绒毛;中肠黏膜上皮最发达,形成的褶皱细长且连接成网状,单层柱状上皮与复层扁平上皮交替分布。观察结果表明,刀鲚消化系统具有典型肉食性鱼类特征。     

运动对小鼠肠道菌群的影响

目的利用PCR-DGGE方法比较运动小鼠和正常小鼠肠道菌群的结构和数量变化,研究运动对肠道菌群的影响。方法从5只正常Balb/c小鼠和5只运动C57BL/6J小鼠的粪便中提取细菌基因组总DNA,用聚合酶链式反应—变性梯度凝胶电泳(PCR-DGGE)的方法获得小鼠肠道菌群分布图谱,对图谱进行相似性、多样性以及优势条带的序列分析。结果 PCR-DGGE结果图显示运动小鼠肠道菌群的多样性明显增加,优势菌群发生转变,序列分析表明变形菌门明显减少,厚壁菌门成为优势菌型。结论运动会对小鼠肠道菌群产生影响。     

长江安庆段刀鲚寄生线虫群落结构及感染特征

为理清刀鲚(Coilia nasus)洄游时间与体内寄生线虫的关系,于2018年4—7月在长江安庆段调查了刀鲚寄生线虫的感染情况及群落结构。结果显示,刀鲚线虫感染率为96.0%,平均感染强度为8.06±7.26,平均感染丰度为7.74±7.29。经ITS分子标记共鉴定出7种线虫,其中异尖属2种,宫脂属4种,针晶蛔属1种,均为海水性寄生虫,可作为刀鲚有海水生活经历的生物标记。其中派氏异尖线虫(Anisakis pegreffii)的感染率最高,为84.5%,其次为内弯宫脂线虫(Hysterothylacium aduncum)(31.0%),二者的感染强度和感染丰度也较高,其中平均感染强度分别为6.40±6.08和2.81±2.49,平均感染丰度分别为5.41±6.05和0.87±1.90;派氏异尖线虫感染丰度和感染强度在洄游早期呈上升趋势,到6月下旬稍有降低但不显著(P>0.05),内弯宫脂线虫感染丰度和强度则均表现为下降趋势。刀鲚寄生线虫的群落结构在不同洄游时间也有所差异,其中刀鲚感染线虫的平均物种丰富度和Brillouin多样性指数均呈下降趋势,但优势物种一直是派氏异尖线虫,...     

Spatial heterogeneity of bacterial community structure in the sediments of the Pearl River estuary

Denaturing gradient gel electrophoresis (DGGE) and multivariate statistical analytical methods were applied to investigate the spatial variation of bacterial community structure in the Pearl River estuary sediment and to address the relationship between microbial community composition and bottom water chemistry in ten different stations. Preliminary results of sequencing analysis of the excised DGGE bands suggested that α-Proteobacteria, γ-Proteobacteria, Acidobacteria and Actinobacteria were the dominant bacterial groups in the Pearl River estuary sediment. Results of multidimensional scaling analysis of these field data suggested that the composition of bacterial communities varied with sampling sites. Finally, canonical correspondence analysis of the data of environmental variables and bacterial community suggested that bacterial community structure was significantly influenced by the change of environmental variables (total phosphorus, nitrite, ammonium, dissolved oxygen, pH and salinity).     

长江刀鲚生殖洄游期间脂肪酸组成及含量变化分析

为探究长江刀鲚生殖洄游过程中脂肪酸组成及其含量变化规律,研究选择洄游距离、卵巢发育和规格大小3个影响因子设置梯度,对长江刀鲚肝胰腺、肌肉和卵巢的脂肪酸组成及含量进行实验分析。实验结果显示, 62尾雌性刀鲚3个组织均检测出28种脂肪酸,以单不饱和脂肪酸(MUFA)含量最高,大于各组织总脂肪酸含量的56.23%,各类脂肪酸中的C18:1、C16:0、C16:1、DHA和EPA含量较高,为主要脂肪酸。在生殖洄游过程中,刀鲚肝胰腺总脂肪含量随洄游距离的延长呈上升趋势,从崇明江段的(526.61±38.50) mg/g增加至安庆江段的(587.21±124.72) mg/g,而肌肉和卵巢总脂肪酸含量呈显著下降趋势,分别下降了33.03%和57.09%(P<0.05)。在各体长组中,肌肉总脂肪酸、SFA、MUFA和PUFA含量与体长呈正相关(P<0.05),而肝胰腺和卵巢总脂肪酸及各类脂肪酸含量与体长无显著相关性(P>0.05)。在卵巢发育过程中,刀鲚肝胰腺和肌肉总脂肪酸、MUFA和PUFA含量随卵巢由Ⅱ期发育至Ⅳ期均呈下降趋势,总脂肪酸含量分别减少了47.56%和22.40%,...     

16S rDNA-Based Analysis of Dominant Bacterial Populations Associated with Early Life Stages of Coho Salmon (Oncorhynchus kisutch)

In this study, we used a 16S rDNA–based approach to determine bacterial populations associated with coho salmon (Oncorhynchus kisutch) in its early life stages, highlighting dominant bacteria in the gastrointestinal tract during growth in freshwater. The present article is the first molecular analysis of bacterial communities of coho salmon. Cultivability of the salmon gastrointestinal microbiota was estimated by comparison of direct microscopic counts (using acridine orange) with colony counts (in tryptone soy agar). In general, a low fraction (about 1%) of the microbiota could be recovered as cultivable bacteria. Using DNA extracted directly from individuals belonging to the same lot, bacterial communities present in eggs and gastrointestinal tract of first-feeding fries and juveniles were monitored by polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE). The DGGE profiles revealed simple communities in all stages and exposed changes in bacterial community during growth. Sequencing and phylogenetic analysis of excised DGGE bands revealed the nature of the main bacteria found in each stage. In eggs, the dominant bacteria belonged to β-Proteobacteria (Janthinobacterium and Rhodoferax). During the first feeding stage, the most abundant bacteria in the gastrointestinal tract clustered with γ-Proteobacteria (Shewanella and Aeromonas). In juveniles ranging from 2 to 15 g, prevailing bacteria were Pseudomonas and Aeromonas. To determine the putative origin of dominant Pseudomonas and Aeromonas found in juvenile gastrointestinal tracts, primers for these groups were designed based on sequences retrieved from DGGE gel. Subsequently, samples of the water influent, pelletized feed, and eggs were analyzed by PCR amplification. Only those amplicons obtained from samples of eggs and the water influent presented identical sequences to the dominant bands of DGGE. Overall, our results suggest that a stable microbiota is established after the first feeding stages and its major components could be derived from water and egg epibiota.     

Analysis of Bacterial Community in the Ginseng Soil Using Denaturing Gradient Gel Electrophoresis (DGGE)

The objective of this work was to determine the shifts in the PCR-DGGE profiles of bacterial communities associated with the rhizosphere soil of ginseng at varying age levels. Differences in the dominance of intense DNA bands in the DGGE profile was observed over the age of the plants indicating the fluctuation in the microbial community structure. The bacterial orders of actinomycetales of Actinobacteria and Spingomonadales and Rhizobiales of α-Proteobacteria were predominant in the ginseng soil.     

Molecular characterization of the bacteria adherent to human colorectal mucosa

AIMS: To study large intestinal mucosal bacterial communities by Denaturing Gradient Gel Electrophoresis (DGGE) profiling and sequencing of 16S rRNA gene polymerase chain reaction (PCR) products amplified from DNA extracted from colorectal biopsies taken from healthy individuals. The specific aims were to determine how similar the mucosa-associated bacterial communities are within and between individuals and also to characterize the phylogenetic origin of isolated DGGE bands. METHODS AND RESULTS: Human colorectal biopsies were taken at routine colonoscopy from 33 patients with normal looking mucosa. The DNA was extracted directly from single biopsies and the bacterial 16S rDNA PCR amplified. The PCR products were profiled using DGGE to generate a fingerprint of the dominant members of the bacterial community associated with the biopsy. The reproducibility of this method was high (>98%). Washed and unwashed biopsies gave similar DGGE banding patterns (Median Similarity Coefficient - MSC 96%, InterQuartile Range - IQR 3.0%, n = 5). Adjacent biopsies sampled from the same patient using different forceps gave similar DGGE profiles (MSC 94%, n = 2). Two colorectal biopsies sampled at locations 2-5 cm apart, from each of 18 patients, resulted in very similar profiles (MSC 100%, IQR 2.8%). Biopsies sampled from different locations within the large intestine of the same patient also gave similar DGGE profiles (MSC 98% IQR 3.3%n = 6). Although all patients (n = 33) gave different DGGE profiles, some similarity (c. 34%) was observed between profiles obtained from 15 patients arbitrarily selected. 35 DGGE bands were excised and sequenced. Many were found to be most closely related to uncultured bacterial sequence entries in the Genbank database. Others belonged to typical gut bacterial genera including Bacteroides, Ruminococcus, Faecalibacterium and Clostridium. CONCLUSIONS: Bacterial communities adherent to colorectal mucosa within a normal patient show little variation; in contrast, mucosal bacterial communities sampled from different patients with normal colorectal mucosa show a high degree of variation. SIGNIFICANCE AND IMPACT OF THE STUDY: This research demonstrates that DGGE profiling of 16S rRNA gene PCR products amplified from DNA extracted directly from mucosal samples offers fresh insight into the bacterial communities that are adherent to colorectal mucosa. These findings are important with respect to further studies on the gastrointestinal tract in health and disease.     

基于PCR-DGGE指纹图谱川纹笛鲷及圆白鲳消化道壁优势菌群结构比较分析

本文采用免培养的16S rDNA梯度凝胶电泳技术(DGGE)对集约化海水网箱养殖川纹笛鲷Lutjanus sebae及圆白鲳Ephippus orbis消化道壁优势菌群结构进行了比较分析。研究结果显示川纹笛鲷及圆白鲳消化道壁存在着大量细菌群落,对DGGE指纹图谱聚类分析表明两种鱼肠道壁及胃壁菌群组成相似度高于50%,其中二者肠道壁细菌组成相似性最高(67%),这些可能与两种鱼养殖在同一水域、摄食相同饵料相关,另外通过软件对DGGE指纹带谱相对丰度分析表明同种鱼肠道壁及胃壁具有相同最大优势菌群。同时,两种鱼消化道壁之间在细菌多样性及相对丰度上亦存在明显区别,圆白鲳消化道壁细菌多样性要高于川纹笛鲷,这可能归因于川纹笛鲷与圆白鲳在天然环境中栖息地的差异性。本研究通过首次建立不同海水鱼消化道壁16S rDNA-DGGE指纹图谱及比较分析,为澄清海水鱼消化道壁微生物区系奠定基础。     

三峡库区短颌鲚年龄和生长特性的研究

为调查短颌鲚(Coilia nasus)在三峡库区的资源现状,自2019年7—12月于三峡库区的长江一级支流澎溪河、梅溪河、草堂河、大宁河共采集短颌鲚样本459尾,用以分析三峡库区现有短颌鲚的年龄结构及生长特征.研究结果显示,三峡库区短颌鲚群体由1—6龄组成,1龄即可达性成熟,群体中1—3龄个体占比90.85％;群体平...