Effect of intraperitoneal injection of immunostimulatory substances on allochthonous gut microbiota of Atlantic salmon (Salmo salar L.) determined using denaturing gradient gel electrophoresis

The allochthonous microbiota in the proximal and distal intestine was investigated in three groups of Atlantic salmon (Salmo salar L.) fed a commercial diet and intraperitoneally injected with (a) phosphate‐buffered saline (control), (b) lipopolysaccharide (LPS) from the fish pathogenic bacteria, Aeromonas salmonicida ssp. salmonicida, and (c) laminaran [β‐(1,3)‐d ‐glucan]. Denaturing gradient gel electrophoresis (DGGE) of the hyper variable V3 region was used to present the microbiota in different experimental groups. Sequencing and phylogenetic analysis of excised DGGE bands suggested that an intraperitoneal injection of LPS from A. salmonicida affects the allochthonous gut bacteria of Atlantic salmon to some extent, as Aeromonas enteropelogenes, Aeromonas veroni, Psychrobacter, Lactobacillus letvazi, Lactobacillus satsumensis, Pantoea, swine manure bacterium and several uncultured bacteria were unique for this group. On the other hand, the bacterial diversity of the allochthonous microbiota did not seem to be affected by injection of β‐(1,3)‐d ‐glucan. Sequences of this experimental group were most closely related to A. enteropelogenes, uncultured Escherichia and Lactobacillus aviarius ssp. aviarius.     

高通量测序分析凡纳滨对虾育苗期水体菌群结构特征

为了解凡纳滨对虾育苗期正常苗池水体菌群结构和多样性变化,实验采用Illumina MiSeq高通量测序水体细菌16S rDNA V4区,比较幼体在无节V期(N5)、溞状Ⅱ期(Z2)、糠虾Ⅰ期(M1)与仔虾Ⅰ期(P1)时的水体菌群特征。结果显示,水体菌群Shannon多样性指数随幼体发育先降后升,而Pielou均匀度指数除在N5时稍高外,在其余阶段都很接近。育苗期间,变形菌门为水体绝对优势菌群(50.0%~88.0%),尤其红杆菌科从N5到P1阶段分别占24.2%、61.6%、43.3%和51.8%;拟杆菌门在各期丰度均9.7%,其中腐螺旋菌科以5.3%~16.9%优势分布于后3个阶段;放线菌门在后3个期都为优势类群,并以微杆菌科为主,丰度分别达5.6%、37.6%和10.8%;而疣微菌门只在P1时表现为优势(5.3%)。苗池水体核心菌属主要由洛克氏菌、海命菌、栖东海菌、亚硫酸杆菌和2个分类未定属构成。可见水体菌群呈高度多样而动态变化,且N5菌群明显区别于后3个阶段。育苗期间水体主要环境因子总体波动较小,对优势操作分类单元(OTU)丰度分布无显著影响。研究表明,凡纳滨对虾健康苗池水体菌群随幼体发育和饵料投喂而明显更替,其中核心菌群的潜在功能对维持水体生态系统稳定起重要作用,可为对虾育苗益生菌与健康苗池指示菌的筛选提供依据。     

Identification of the adherent microbiota on the gills and skin of poly‐cultured gibel carp (Carassius auratus gibelio) and bluntnose black bream (Megalobrama amblycephala Yih)

PCR‐denaturing gradient gel electrophoresis (DGGE) was applied to analyse the microbial community attached to the gills and skin of poly‐cultured gibel carp (Carassius auratus gibelio) and bluntnose black bream (Megalobrama amblycephala Yih) and compare these results with those detected in the rearing water. The microbiota discussed included bacteria, fungi and a specific bacterial taxa of actinomycetes was also analysed. Proteobacteria, Firmicutes, Actinobacteria, Cyanobacteria, Ascomycota, Basidiomycota and some unclassified microbiota were identified. Based on our results, we concluded that: (1) the adherent bacterial/fungal communities on the gills and skin were different from those in the rearing water, (2) the bacterial/fungal diversities on fish gills were lower than that on fish skin, (3) the adherent bacterial/fungal communities on gill and skin of gibel carp were different from that of bluntnose black bream and (4) the adherent actinomycetal community showed certain similarity between the skin of different hosts. Based on our conclusions, we suggested that the topic investigated in the present study merits further investigations.     

不同月份养殖草鱼幼鱼消化道微生物群落动态变化研究

为了评估草鱼消化道微生物群落结构在草鱼食性转化后的变动情况,实验对食性转换完成后的草鱼消化道微生物群落结构进行了PCR-DGGE分析。结果显示,不同月份采集草鱼消化道微生物群落结构存在明显差异(Monte Carlo检验,A:F=3.41,P=0.002;B:F=3.58,P=0.002),而被广泛认为影响宿主消化道微生物群落结构的宿主大小、丰满度等因素则在短期内并没有发现会对草鱼消化道微生物群落结构产生影响。实验表明,短期内环境因素可能是造成草鱼消化道微生物群落结构变动的主要因素。本实验结果将为深入阐述草鱼消化道微生物群落结构的变化规律、消化道益生菌的作用机理、消化道微生物群落结构变动与草鱼疾病的关系等问题积累基础材料。     

许氏平鲉仔鱼、稚鱼、幼鱼肠道微生物群结构特征

采集常规饲养的健康许氏平鲉1日龄仔鱼,9、20日龄稚鱼,54、95日龄幼鱼,运用Illumina MiSeq PE300平台,对其肠道微生物的16S rDNA V3~V4可变区进行高通量测序,共获得1039个运算分类单位、667个属。在丰度高于1%的菌属水平上,野生鱼肠道菌群丰度的平均值(86%)远高于幼鱼时期的平均值(73.5%)。不动杆菌属为仔鱼、稚鱼、幼鱼肠道中最优势菌属(丰度为26.2%~45.3%);短芽孢杆菌属次之。结合共有菌属分析获得11个核心菌群,芽孢杆菌属、短芽孢杆菌属、不动杆菌属、根瘤菌属等的丰度随仔鱼、稚鱼、幼鱼生长先升后降,均在54日龄时丰度最高;弧菌属则呈现相反的趋势(0.37%~12.41%),54日龄时丰度最低。仔鱼、稚鱼、幼鱼与野生个体肠道仅有14个共有菌属,短芽孢杆菌属、Massilia的最小丰度值分别是野生个体肠道中最大丰度值的9倍和32倍,差异明显。本研究揭示了许氏平鲉仔鱼、稚鱼、幼鱼肠道菌群结构特征及其发生、发育的过程,为苗种繁育阶段肠道生理健康调控提供理论依据。     

Structural characteristics and succession of intestinal microbiota for Paralichthys olivaceus during the early life stage

The research of intestinal microflora structures for Paralichthys olivaceus larvae and juveniles will help us to master the ontogeny and developmental colonization of microflora during the larval rearing stage. In this study, we sequenced the total bacterial genomic DNA in larval and juvenile guts with an Illumina MiSeq PE300 system, and analysed the structural characteristics of these microbiota, feed and rearing water in live and formulated feeding periods. The structure of gut microbiota was gradually similar to those in wild P. olivaceus at the phylum level and the newly hatched ones with the growth, according to the distribution and abundance of intestinal microbiota. And the colonized rule of main microbial species in guts was decreased initially and then increased during the larvae and juveniles stage. Meantime, the core microflora of this study were obtained through the analysis of shared and dominant species, which included Bacteroides, Bacillus, Enterococcus, Lactobacillus, Lactococcus, Escherichia_Shigella, Acinetobacter, Pseudomonas, Vibrio, Nitrosomonas, and Glaciecola. The correlation analysis of microbiota between intestines and environmental factors suggested that microflora in feed and water could affect the distribution of larval and juvenile gut microbiota. Moreover, many species of Acinetobacter, Pseudomonas and Vibrio are treated as important potential pathogens in aquaculture. These all pointed out the microbial quality of feed and rearing water should be strictly controlled in fish breeding and farming, and supply theoretical bases for screening the native probiotics to artificially regulate the gut microbiota.     

主养草鱼池塘三种混养模式下鱼类肠道菌群PCR-DGGE比较

消化道微生物在宿主生长、营养和健康等方面起到重要的作用.本实验采用基于PCR扩增的变性梯度凝胶电泳(PCR-DGGE)技术比较研究了主养草鱼(Ctenopharyngodon idellus)池塘三种不同混养模式下的鱼类肠道菌群差异.结果表明,三种混养模式下同种鱼的生长率出现显著性差异(P＜0.05),而肠道细菌16SrDNA V3区特征片段PCR-DGGE指纹分析显示草鱼的肠道菌群相似性较高(＞42.2％);投喂配合饲料的草鱼与摄食浮游生物的鲢(Hypophthalmichthys molitrix)、鳙(Aristichthys nobilis和匙吻鲟(Polyodon spathula)肠道菌群结构相差最大(＜19％),鲢和鳙肠道菌群相似性较高(＞41.6％),除模式Ⅱ鳙的肠道和匙吻鲟的胃菌群具有较高的相似性(＞50.3％)外,匙吻鲟的消化道菌群和鲢鳙的相似性低.实验共回收测序了14条特定DGGE条带中的DNA片段,并进行系统进化分析,结果显示,这14条条带分别归属于4个细菌类群:变形细菌门(Proteobacteria),拟杆菌门(Bacteroidetes),厚壁菌门(Firmicutes)和梭杆菌门(Fusobacteria).研究结果为鱼类混养模式的优化,饲料研发和鱼病防治提供了基础参考资料.     

(微)塑料污染对土壤生态系统的影响:进展与思考

塑料已经成为现代社会不可或缺的产品而被广泛应用,塑料污染也成了一个全球性的环境污染问题。近年来,土壤塑料污染的问题也开始受到关注。本文针对近几年来国内外关于塑料污染对土壤生态系统的影响进行综述,主要包括以下几个方面:(1)微塑料对土壤物理化学性质的影响;(2)微塑料对土壤微生物群落的影响;(3)微塑料与土壤动物的相互作用。最后,本文对未来关于土壤微塑料研究的重点方向进行了展望。     

适钙植物报春苣苔根际土壤微生物群落结构

为解析喀斯特适钙植物报春苣苔根际微生物群落结构的特点,采用高通量测序16S rRNA V4和ITS1区序列,比较分析了适钙植物报春苣苔根际/非根际土壤微生物群落结构差异,并利用CCA及斯皮尔曼相关系数分析了土壤可溶性钙含量和报春苣苔根际/非根际土壤微生物群落之间的关系。结果表明：报春苣苔根际土壤微生物群落包含41个细菌门及6个真菌门。细菌中优势菌门为变形菌门（Proteobacteria）、放线菌门（Actinobacteria）及酸杆菌门（Acidobacteria）,占64%以上;真菌主要为子囊菌门（Ascomycota）、担子菌门（Basidiomycota）和接合菌门（Zygomycota）,占99%以上。相比非根际土壤,在根际土壤中酸杆菌门菌群丰度显著上升,而变形菌门和芽单胞菌门丰度显著下降,生长受到抑制。报春苣苔的根际和非根际土壤微生物群落非常丰富,包含569个属的细菌及276个属的真菌。报春苣苔根际细菌的Shannon、Chao1和ACE等α-多样性指数显著高于非根际,表明报春苣苔根际细菌菌群种类数量高于非根际土壤;而报春苣苔根际真菌的PD_whole_tree指数显著低于非根际,表明报春苣苔根际真菌群落的群体进化低于非根际。土壤可溶性钙对于微生物群落结构的解释量仅为16.93%,且蒙特卡罗置换检验表明土壤可溶钙和物种分布显著不相关性;只有Hydrogenedentes门细菌与土壤可溶性钙显著正相关。报春苣苔根际/非根际土壤微生物群落之间差异显著,且两者与土壤可溶性钙的相关性有限;报春苣苔根际土壤微生物的物种组成丰富,且报春苣苔根系有利于根际土壤细菌群落多样性。     

K94复合活菌剂对蛋鸡产蛋性能、饲料消化率和盲肠菌群的影响

240只60周龄的海赛克斯褐壳蛋鸡用于作K94的应用效果试验。试验开始2周后，试验组（基础日粮中添加1％K94）产蛋率提高7．85％，对照组（添加无活菌的1％K94）则降低2．34％。日粮中未添加维生素后．试验组产蛋率下降28．15％，对照组则下降38．49％。重新添加维生素后，试验组和对照组产蛋率都迅速回升，但试验组回升速度比对照组快。整个试验期（9周），试验组产蛋率和枚蛋重都明显高于对照组；料蛋比明显比对照组低。饲喂K949周后，与对照组比较，试验组采食量增加8．01％，饲料干物质（DM）和粗蛋白（CP）消化分别提高14．79％和18．49％；盲肠内容物中大肠杆菌明显减少，而厌氧乳杆菌、梭状芽胞杆菌以及双歧杆菌数量明显增多。结果表明，K94能明显改善盲肠微生态环境，提高饲料消化率，从而提高蛋鸡的产蛋性能，改善日粮中维生素不足对产蛋性能的影响。