微型生物群落eDNA-PFU监测

微型生物群落在水生态系统中起着至关重要的作用, 是水生态系统食物链的关键环节, 其结构和功能可以很好地反映不同生境的水质特征。《水质 微型生物群落监测 PFU法》是我国首个生物监测国家标准, 被证明是一种快速、经济、准确的监测方法。然而, 由于PFU法对专业知识要求较高、费时耗力且不易标准化, 严重阻碍了该方法进一步的推广与应用。近年来, 环境DNA(eDNA)技术的快速发展突破了微型生物形态鉴定的瓶颈, 能高效、准确地鉴定出水环境中包括微型生物在内的各种水生生物。为此, 文章提出微型生物群落eDNA-PFU法, 即基于eDNA技术改进的PFU法, 对微型生物群落进行监测。为了对eDNA-PFU法进行验证, 文章针对武汉东湖水体中的原生动物群落开展了预实验, 结果表明eDNA-PFU法较传统PFU法覆盖度与灵敏度均更高, 可以真实、全面、准确地揭示水体中微型生物群落结构特征。因此, 基于eDNA-PFU法的微型生物群落监测有望为湖泊、河流、水库等水体的生态考核提供新一代的生物监测标准。     

流式细胞术在水体微型生物分子生物学研究中的应用

流式细胞术(FCM)是利用流式细胞仪对微小生物颗粒物的多种物理、生物学特性进行定量,并对特定细胞群体进行分选的分析测量技术。流式细胞仪是当代激光、流体力学、光学和电子计算机等学科技术高度发展的产物,是生命科学研究领域中先进的仪器之一。FCM最早应用于海洋生物学是在80年代中期,特别是1988年发现原绿球藻后[1],FCM成为研究水生微型生物(包括超微型生物)的重要手段。近年来,该领域的有关研究已成为海洋生命科学的研究热点,并从最初区分、记数不同微型浮游植物种群,发展到现在DNA分析和以荧光分子探针为辅助手段的系统发育分析…     

海洋微型底栖生物的多样性与地理分布

海洋微型底栖生物是指生活于海洋沉积物中及表面的所有单细胞原核和真核微型生物,包括原核微生物、真核微藻及原生动物等光合自养和异养的类群.与水体相比,海洋底栖生境孕育了形态和功能多样性更高的微型生物,在陆架浅海单位体积沉积物中其丰度较之水体中同类生物高一至几个数量级,而深海则孕育着特殊进化环境下新奇多样且数量庞大的微型底栖生物,是维持海洋生物多样性、海洋生态系统结构和功能不可或缺的部分.迄今对于微型生物是全球性还是限定性分布一直存在争议,对其解答受到分类研究欠缺及采样不足的制约.分子生物学从正反两方面提供了理论依据,但无法得出一个普遍接受的观点.海洋微型底栖生物的多样性研究侧重于物种多样性及群落结构与分子多样性,较为显著的进展体现在原核微生物的分子多样性及底栖真核微藻的物种多样性研究,对于海洋底栖原生动物的多样性研究则相较滞后.本文综述了国内外对海洋微型底栖生物各主要类群的分类学和多样性研究进展,探讨了各类群在全球的潜在物种多样性,并就我国未来加强海洋微型底栖生物多样性构成、分布与变动及驱动变化的因子以及底栖微食物网的研究提出了建议.     

生物修复的研究和应用现状及发展前景

综述了生物修复的研究和应用现状及发展前景。内容涉及生物修复的研究方法和手段、生物修复在土壤和水体中的应用、生物修复的发展前景等。     

珠江浮游真核微型生物分子多样性及其与水环境的关系

文章分别在丰水期和枯水期于珠江广州段和西江广东段43个样点采集水样, 基于末端限制性长度多态性分析, 研究了珠三角地区珠江水体的浮游真核微型生物的分子多样性及其群落结构和水体理化因子的关系。结果表明, 西江广东段及珠江广州段水体氮和磷严重超标, 水质状况堪忧。枯水期样本浮游真核微型生物香农威尔多样性指数普遍高于丰水期样本, 西江广东段样本浮游真核微型生物多样性指数普遍高于珠江广州段样本。不同水期, 不同区域的浮游真核微型生物群落结构均存在显著差异。珠江样点的浮游真核微型生物群落结构均与化学需氧量、高锰酸盐指数、氨氮、总氮、总磷高度相关; 但不同水期、不同区域的浮游真核微型生物群落结构与理化因子相关系数有所差异。此外, 分别筛选出了1个、8个末端限制性片段代表了潜在的敏感(Cystobasidium sp.或Protostelium nocturnum)、耐污(Acanthamoeba hatchetti、Babesia bicornis、Blastocystis sp.、Botryosphaerella sudetica、Candida caryicola、Coccomyxa simplex、Cryptomonas ovata、Filos agilis、Stenophora robusta、Sulfonecta uniserialis、Theileria sp.等)物种/类群。     

流式细胞术在富营养淡水湖泊微型浮游植物细胞中的应用

应用流式细胞术(FCM)对一个富营养化淡水湖泊表、底层微型浮游植物细胞进行了初步研究。研究结果表明：流式细胞术可快速、多参数区分3种不同类群微型浮游植物。微型浮游植物细胞在表、底层占50μm以下微型颗粒物数量比例分别为21．08％、17．87％,在不同水层,微型浮游植物的优势类群及数量也不同。流式细胞术大大提高了淡水微型浮游生物研究监测水平。     

用微型生物群落评价常德市水系的研究

本文应用PFU法对常德水系四季微型生物群落的结构和功能进行了分析。根据MacArthur-Wilson平衡模型提出了3个功能参数,测定了微型生物的2个结构参数。同步进行了16个站四季的水质分析,提出化学综合污染指数(P)。经统计学处理,P和微型生物群落的HI、S_(eq)、G、T_(90％)等指数均呈显著性相关。物种多样性(d)也在一定程度上反映出城市水体的不同污染程度。再次证明PFU微型生物监测方法的科学性,为生物监测提供了快速、经济、正确的新方法。     

海洋有害赤潮的生物防治对策

该文综述了海洋有害赤潮治理的生物方法。细菌、寄生虫和病毒等微型生物是天然水体中具有调节藻类种群动态平衡的重要潜在因子,它们的繁殖速度非常快,并具有宿主的专一性,是一种非常有潜质的赤潮调控因子。根据生态系统中的食物链关系,通过引入赤潮生物的天敌来防治赤潮也是一种治理方法。浮游动物和一些滤食性的贝类在水体中与赤潮生物共存,并能以这些赤潮生物为食,是具有明显赤潮治理功效的生物控制剂。另一方面,利用大型海藻与微藻间的相生相克和营养竞争等作用在养殖区域内进行赤潮的防控被认为具有较强的理论和实践意义。该文详细介绍了上述方法的原理、可行性以及局限性,并就今后的研究方向进行了展望。     

水污染的微型生物监测

一切污染物最终都要流入水体。对水体进行生物监测是控制水污染的关键。本文从公害评价的基本原理出发,对已成为我国国家标准《水质-微型生物群落监测-PFU法》的生态学基础、方法的改进和创新,在国内的推广、应用及其标准化的过程作了扼要介绍。对新方法提出的5个问题作了解答。     

利用拟杆菌分子标记物对粪便污染溯源的研究进展

拟杆菌(Bacteroidales)是粪便污染的主要标志之一,近年来被广泛应用在水体微生物溯源领域,并展现了良好的应用前景.拟杆菌微生物溯源的优势在于不需要纯培养过程,直接根据拟杆菌具有的宿主特异性生物标记引物设计,通过检测水体中拟杆菌生物标记的存在情况来判断粪便污染来源.本文从人、猪、反刍动物等拟杆菌特异性生物标记的发现、引物设计以及其应用综述了拟杆菌在微生物溯源中的研究进展,指出了应用过程中的局限性.本文还对拟杆菌在溯源中的应用进行了展望.提出了寻找新的宿主特异性拟杆菌生物标记是研究的主要方向,在应用研究中应注重与其他溯源方法相结合以使溯源结果更加准确.     

Bacterial microbiota assemblage in Aedes albopictus mosquitoes and its impacts on larval development

Interactions between bacterial microbiota and mosquitoes play an important role in mosquitoes’ capacity to transmit pathogens. However, microbiota assemblages within mosquitoes and the impact of microbiota in environments on mosquito development and survival remain unclear. This study examined microbiota assemblages and the effects of aquatic environment microbiota on the larval development of the Aedes albopictus mosquito, an important dengue virus vector. Life table studies have found that reducing bacterial load in natural aquatic habitats through water filtering and treatment with antibiotics significantly reduced the larva‐to‐adult emergence rate. This finding was consistent in two types of larval habitats examined—discarded tires and flowerpots, suggesting that bacteria play a crucial role in larval development. Pyrosequencing of the bacterial 16S rRNA gene was used to determine the diversity of bacterial communities in larval habitats and the resulting numbers of mosquitoes under both laboratory and field conditions. The microbiota profiling identified common shared bacteria among samples from different years; further studies are needed to determine whether these bacteria represent a core microbiota. The highest microbiota diversity was found in aquatic habitats, followed by mosquito larvae, and the lowest in adult mosquitoes. Mosquito larvae ingested their bacterial microbiota and nutrients from aquatic habitats of high microbiota diversity. Taken together, the results support the observation that Ae. albopictus larvae are able to utilize diverse bacteria from aquatic habitats and that live bacteria from aquatic habitats play an important role in larval mosquito development and survival. These findings provide new insights into bacteria's role in mosquito larval ecology.     

Development and laboratory-scale testing of a fully automated online flow cytometer for drinking water analysis

Accurate and sensitive online detection tools would benefit both fundamental research and practical applications in aquatic microbiology. Here, we describe the development and testing of an online flow cytometer (FCM), with a specific use foreseen in the field of drinking water microbiology. The system incorporated fully automated sampling and fluorescent labeling of bacterial nucleic acids with analysis at 5-min intervals for periods in excess of 24 h. The laboratory scale testing showed sensitive detection (< 5% error) of bacteria over a broad concentration range (1 × 10(3) -1 × 10(6) cells mL(-1) ) and particularly the ability to track both gradual changes and dramatic events in water samples. The system was tested with bacterial pure cultures as well as indigenous microbial communities from natural water samples. Moreover, we demonstrated the possibility of using either a single fluorescent dye (e.g., SYBR Green I) or a combination of two dyes (SYBR Green I and Propidium Iodide), thus broadening the application possibilities of the system. The online FCM approach described herein has considerable potential for routine and continuous monitoring of drinking water, optimization of specific drinking water processes such as biofiltration or disinfection, as well as aquatic microbiology research in general.     

The effects of taxonomy,diet, and ecology on the microbiota of riverine macroinvertebrates

Freshwater macroinvertebrates play key ecological roles in riverine food webs, such as the transfer of nutrients to consumers and decomposition of organic matter. Although local habitat quality drives macroinvertebrate diversity and abundance, little is known about their microbiota. In most animals, the microbiota provides benefits, such as increasing the rate at which nutrients are metabolized, facilitating immune system development, and defending against pathogenic attack. Our objectives were to identify the bacteria within aquatic invertebrates and determine whether their composition varied with taxonomy, habitat, diet, and time of sample collection. In 2016 and 2017, we collected 264 aquatic invertebrates from the mainstem Saint John (Wolastoq) River in New Brunswick, Canada, representing 15 orders. We then amplified the V3‐V4 hypervariable region of the 16S rRNA gene within each individual, which revealed nearly 20,000 bacterial operational taxonomic units (OTUs). The microbiota across all aquatic invertebrates were dominated by Proteobacteria (69.25% of the total sequence reads), but they differed significantly in beta diversity, both among host invertebrate taxa (genus‐, family‐, and order‐levels) and temporally. In contrast to previous work, we observed no microbiota differences among functional feeding groups or traditional feeding habits, and neither water velocity nor microhabitat type structured microbiota variability. Our findings suggest that host invertebrate taxonomy was the most important factor in modulating the composition of the microbiota, likely through a combination of vertical and horizontal bacterial transmission, and evolutionary processes. This is one of the most comprehensive studies of freshwater invertebrate microbiota to date, and it underscores the need for future studies of invertebrate microbiota evolution and linkages to environmental bacteria and physico‐chemical conditions.     

Flow cytometric analysis of fine particles in a eutrophic lake.

Fine particles play an important role, not only in aquatic biogeochemical processing but also in the distribution, transfer and transformation of pollutants in the aquatic environment. Flow cytometry, widely used in biomedical research, allows fast counting and optical analysis of individual particles. Organic autotrophic particles contain naturally fluorescing pigments, such as chlorophyll and phycoerythrin. Different populations have different sizes and pigments. They also have different ratios of pigments. In general, side angle scatter (SSC) is related to the size, shape and refractive index of particles. When a 488 nm wavelength was used to excite chlorophyll and phycoerythrin fluorescence, the pigments of organic autotrophic particles emitted red and orange light. Fine particles were detected by flow cytometry (FCM) in the southern part of a eutrophic lake in winter. We found that organic autotrophic particles belonged to three populations, which represented only 15.89% of total fine particles. Organic non-living particles and inorganic particles represented the greater part (84.11%) of total fine particles. This study also demonstrated that flow cytometry is well suited to the dynamic monitoring and analysis of natural water aquatic particles that were difficult to study with traditional methods.     

Genetic and Physiological Characterization of the Intestinal Bacterial Microbiota of Bluegill (Lepomis macrochirus) with Three Different Feeding Habits

Bluegill (Lepomis macrochirus) in Lake Biwa, Japan, feed on benthic invertebrates (benthivorous type), aquatic plants (herbivorous type), and zooplankton (planktivorous type). To evaluate the effect of food on intestinal bacterial microbiota, we characterized and compared the intestinal microbiota of these three types of bluegill in terms of community-level physiological profile (CLPP) and genetic structure. The CLPP was analyzed using Biolog MicroPlates (Biolog, Inc., Hayward, CA, USA), and multivariate analysis of variance revealed that the CLPP of intestinal microbiota differed significantly between any pairs of the three types of bluegill. The genetic profiles were analyzed by temperature gradient gel electrophoresis of polymerase chain reaction (PCR)-amplified 16S rDNA fragments, and multidimensional scaling indicated the existence of specific intestinal bacterial structures for both the benthivorous and the planktivorous types. These results suggest that the host's feeding habit can be one factor controlling the intestinal microbiota of fish in the natural environment.     

基于性别二分类的青海天峻藏民肠道菌群差异分析

目的对青海省天峻县的藏族牧民健康志愿者肠道菌群进行研究,探讨青海藏民肠道菌群结构及性别对肠道菌群组成的影响。方法以28例藏族牧民志愿者的粪便样品作为研究对象,应用基于16SrDNA V3+V4可变区的高通量测序技术测定肠道菌群组成及核心菌群;通过db-RDA和菌群多样性分析,探索性别因素对肠道菌群结构的影响。结果在门水平上,厚壁菌门和拟杆菌门是青海藏族志愿者的优势菌群。在属水平上,优势菌属是普氏菌属、Lachnospiracea_incertae_sedis、Faecalibacterium和拟杆菌属;特有优势菌属是Lachnospiracea_incertae_sedis。db-RDA结果显示性别因素对肠道微生物群结构有一定分离趋势,多样性结果进一步证实性别差异显著影响菌群结构。男性和女性存在12种差异菌群,包括普氏菌属、Acidaminobacter属等。结论性别差异对青海藏民肠道菌群影响显著。     

宿主遗传因素对肠道菌群的影响

随着高通量测序技术的发展,人们逐渐认识到肠道菌群与人类的健康和疾病密切相关,并发现肠道菌群受很多因素的影响。除了研究传统饮食和药物对肠道菌群的改变外,近年来,科学家也开始注重遗传因素在塑造肠道菌群中的作用。遗传因素可决定宿主的饮食偏好、肠道的生理结构、肠道屏障功能和免疫功能等,而这些都直接与肠道菌群相互作用,参与肠道微生态平衡的构建和稳定。因此,在研究肠道菌群与疾病发生相关性的过程中也需要考虑遗传因素的重要性。随着基因敲除、无菌小鼠和菌群移植等实验技术的革新,以及主成分分析、数量性状基因座和全基因组关联性分析等大数据分析手段的提高,科学家能够深入研究宿主遗传基因与肠道菌群之间的关联性,从而证明宿主遗传基因在塑造肠道微生态的过程中具有重要作用。本文将首先简述肠道菌群与疾病发生之间可能存在的联系,然后从多方面综述遗传因素对肠道菌群的影响及主要的研究进展,从而为今后该领域的深入研究提供重要的指导,也为今后预防和治疗疾病提供新思路和新方法。     

Health benefits of microalgae and their microbiomes

Microalgae comprise a phylogenetically very diverse group of photosynthetic unicellular pro- and eukaryotic organisms growing in marine and other aquatic environments. While they are well explored for the generation of biofuels, their potential as a source of antimicrobial and prebiotic substances have recently received increasing interest. Within this framework, microalgae may offer solutions to the societal challenge we face, concerning the lack of antibiotics treating the growing level of antimicrobial resistant bacteria and fungi in clinical settings. While the vast majority of microalgae and their associated microbiota remain unstudied, they may be a fascinating and rewarding source for novel and more sustainable antimicrobials and alternative molecules and compounds. In this review, we present an overview of the current knowledge on health benefits of microalgae and their associated microbiota. Finally, we describe remaining issues and limitation, and suggest several promising research potentials that should be given attention.     

Interactions between commensal bacteria and viral infection: insights for viral disease control in farmed animals

Viral diseases cause serious economic loss in farmed animals industry. However, the efficacy of remedies for viral infection in farmed animals is limited, and treatment strategies are generally lacking for aquatic animals. Interactions of commensal microbiota and viral infection have been studied in recent years, demonstrating a third player in the interaction between hosts and viruses. Here, we discuss recent developments in the research of interactions between commensal bacteria and viral infection,including both promotion and inhibition effect of commensal bacteria on viral pathogenesis, as well as the impact of viral infection on commensal microbiota. The antiviral effect of commensal bacteria is mostly achieved through priming or regulation of the host immune responses, involving differential microbial components and host signaling pathways, and gives rise to various antiviral probiotics. Moreover, we summarize studies related to the interaction between commensal bacteria and viral infection in farmed animals, including pigs, chickens, fish and invertebrate species. Further studies in this area will deepen our understanding of antiviral immunity of farmed animals in the context of commensal microbiota, and promote the development of novel strategies for treatment of viral diseases in farmed animals.     

Metabolites from freshwater aquatic microalgae and fungi as potential natural pesticides

Microorganisms are recognized worldwide as the major source of secondary metabolites with mega diverse structures and promissory biological activities. However, as yet many of them remain little or under-explored like the microbiota from freshwater aquatic ecosystems. In the present review, we undertook a recompilation of metabolites reported with pesticidal properties from microalgae (cyanobacteria and green algae) and fungi, specifically from freshwater aquatic habitats.