常见疾病、抗生素及农药对意大利蜜蜂(Apis mellifera ligustica)肠道微生物多样性的影响研究进展

蜜蜂是对农业生产十分重要的授粉昆虫。蜜蜂肠道微生物与蜜蜂健康有密切关系,但肠道微生物也会受多种外界因素的影响。本文就蜜蜂疾病、抗生素等蜂病治疗药物、农药,以及益生菌的应用等对意大利蜜蜂工蜂肠道微生物影响的研究进行了归纳总结,并对蜜蜂与其肠道菌关系研究进行了展望。     

洞察景观环境影响蜜蜂之新视角: 肠道微生物

作为生态服务提供者的传粉蜜蜂与景观生态息息相关, 而以农田为主的景观组成显著降低了传粉蜜蜂的多样性。目前调查研究显示, 农田的扩张与蜜蜂多样性下降相关, 且农药残留对蜜蜂损害严重。景观中的开花植物决定了蜜蜂的食物(营养)组成, 其中花粉蛋白含量与蜜蜂的生长发育紧密相关。尽管研究已证实景观环境会显著影响蜜蜂蜂群的发展和个体的生长繁殖能力, 但未来还需要加强景观组成变化直接作用于蜜蜂的机制研究。另一方面, 大量研究表明蜜蜂肠道共生菌是影响宿主健康的重要因素: 可促进宿主吸收营养和抵抗病原菌。作为传粉者, 蜜蜂接触到的主要外部环境——花粉和花蜜都含有特殊的微生物, 很多研究暗示花源微生物是蜜蜂肠道菌来源之一。研究表明景观环境相关的食物(营养)、农药残留以及环境微生物都会显著影响肠道微生物。现有少量的研究证明不同景观的蜜蜂肠道微生物有差异, 景观环境可能通过作用于蜜蜂肠道微生物进而影响蜜蜂健康。然而不同景观环境中的微生物, 尤其是花源微生物和蜜蜂肠道菌之间的关联有待证明。景观对蜜蜂肠道微生物的影响值得研究, 希望可以从肠道菌的视角鉴别对蜜蜂友好的景观环境, 进而指导土地合理利用和蜜蜂保护。     

蜜蜂肠道菌群定殖研究进展

肠道菌群在其宿主健康中发挥着各种各样的重要功能。蜜蜂是高度社会化的昆虫,其肠道菌群与大多数昆虫明显不同,由兼性厌氧和微好氧的细菌组成,具有高度保守性和专门的核心肠道微生物群。近年来的研究表明,蜜蜂肠道微生物群在代谢、免疫功能、生长发育以及保护机体免受病原体侵袭等方面起着重要作用,并已证实肠道微生物在蜜蜂健康和疾病中起着重要作用,肠道微生物群的破坏对蜜蜂健康会产生不利影响。本文综述了蜜蜂肠道菌群的特征及菌群定殖研究进展,介绍了蜜蜂的日龄、群体、季节等对蜜蜂肠道菌群定殖的影响,探讨了宿主的功能和新陈代谢对肠道菌群的影响。     

蜜蜂肠道微生物与其社会性的关系

蜜蜂是一种典型的营群居生活的社会性昆虫,相比独居生活的昆虫,其肠道微生物具有独特的区系结构。这种独特肠道微生物与其社会性之间的关系是一个重要的科学问题。现有研究显示,蜜蜂肠道的优势菌包括9大类群。消化道不同区段的优势菌种类和丰度存在差异。主要表现为前肠种类少、丰度低、后肠种类多、数量大,占了全消化道微生物的99%以上。不同社会分工的蜜蜂肠道微生物区系结构存在差异,肠道微生物会通过影响胰岛素信号的传导、保幼激素和卵黄原蛋白的合成以及蜜蜂抗氧化应激的能力等对蜜蜂的级型分化、社会分工、摄食行为及寿命长短产生调节作用。除此之外,蜜蜂肠道微生物还具有激活免疫、抑制病原菌生长、降解食物、促进养分吸收、解毒、发酵蜂蜜和蜂粮等作用。主要针对蜜蜂肠道微生物的基本特征及其与蜜蜂社会性的关系作一简要综述。     

肠道菌群在蜜蜂健康与疾病中的作用研究进展

蜜蜂是重要的经济昆虫,在农作物授粉和维持生态平衡等方面扮演了重要角色。随着生态环境的逐步恶化,蜜蜂极易受到寄生虫、农药、抗生素和病原微生物等的危害。肠道作为蜜蜂的重要免疫器官,在抵抗外源刺激等方面具有不可替代的作用。而肠道菌群作为肠道的重要组成部分,对蜜蜂的健康有着至关重要的影响。其拥有高度保守和专门的核心微生物群,主要由九大类细菌组成。近年来,有关肠道菌群的研究主要集中在新陈代谢、免疫防御、生长发育等方面。肠道菌群不仅可以帮助宿主消化和制造营养素,还可维持宿主体内能量稳态,其对蛋白质的代谢也使菌群丰度增加。越来越多的证据表明,肠道菌群紊乱会导致蜜蜂健康受损,其多样性对宿主健康和疾病影响甚广。因此,肠道菌群成为近年来微生物学领域研究的焦点。主要综述了蜜蜂属、熊蜂属等主要传粉蜂类肠道菌群的组成及其在健康与疾病中的作用,为深入了解传粉昆虫肠道菌群功能提供参考。     

肠道微生物在动脉粥样硬化发生发展中的作用机制及未来治疗靶点的研究进展

动脉粥样硬化是多种心脑血管疾病的重要病理基础。目前研究发现肠道微生物在动脉粥样硬化的风险因素中起着非常重要的作用。随着研究的深入,人们发现肠道微生物及其代谢产物可能通过影响人体物质代谢、能量的吸收与利用、全身慢性炎症与氧化应激等,参与动脉粥样硬化的发生发展。本文就肠道微生物在动脉粥样硬化的发生发展中作用机制及临床治疗靶点作一简要综述。     

用于蜜蜂和熊蜂肠道微生物分类的细菌16S rRNA数据库优化

蜜蜂和熊蜂是重要的传粉昆虫, 对农业生产及生态平衡的维持具有重要作用。近年来, 研究发现蜜蜂及熊蜂肠道内含有大量微生物, 其组成简单、特异。正常的肠道微生物群落对蜜蜂的生长、激素调节、致病菌抵抗等具有重要作用。随着高通量测序的发展, 研究者们也可快速获得传粉蜂肠道微生物组成, 这给生物多样性和物种保护及蜂类健康等的研究带来了便捷。但是由于蜜蜂和熊蜂肠道微生物群落均由特殊菌种组成, 目前的细菌16S rRNA数据库无法对其进行准确的分类, 并且部分东方蜜蜂(Apis cerana)特有的肠道微生物菌种缺乏16S rRNA序列信息。本文从来源于5个不同省份的东方蜜蜂肠道中分离得到在东方蜜蜂中普遍含有的Apibacter菌属纯菌, 获取其全长16S rRNA序列, 并对目前蜜蜂和熊蜂肠道的5个核心菌种的分类进行了综述, 对其分类和命名进行了修正。根据蜜蜂肠道微生物的明确分类, 在目前常用的SILVA细菌分类数据库基础之上对其进行了命名及分类优化, 并加入东方蜜蜂中普遍含有的Apibacter序列, 从而获得了优化数据库Bee Gut Microbiota-Database (BGM-Db)。通过1组东方蜜峰及1组西方蜜蜂(Apis mellifera)的肠道菌群高通量测序结果, 分析不同数据库的表现, 我们发现相比于SILVA和Ribosomal Database Project (RDP), BGM-Db对蜜蜂肠道16S rRNA高通量测序短序列实现了菌种级别的分类, 分辨率更高。     

肠道微生物和肠-脑轴在肥胖发展中的作用

肥胖和相关代谢综合征已成为全球最突出的健康问题之一,肠道微生物已被证明参与肥胖的发展,并可能对肥胖的发展和其干预治疗等提供重要的见解。最近的研究表明,肠道微生物和大脑的相互作用可能是肥胖的后果或解释因素,肠-脑轴是它们相互作用的联络枢纽。此外,肠道微生物可以通过肠道激素（包括ghrelin）以及迷走神经连接（影响能量消耗和CNS中与饮食行为相关的区域）来影响肠-脑轴,从而改变宿主行为。同时,肠道微生物代谢物和其产物还可以充当信号分子并调节肠内分泌细胞的激素分泌,如GLP1和PYY,从而调节食欲、肠道运动、能量吸收和储存以及能量消耗等摄食相关行为,进而影响肥胖的发展。所以,理解这些信号和激素作用并从药理学方法增强它们,可能为治疗肥胖提供一种重要的途径。     

非人灵长类肠道微生物研究进展与展望

肠道微生物组被誉为动物的“第二套基因组”,与动物的个体发育、营养获取、生理功能、免疫调节等重要活动密切相关。非人灵长类在生态位、社会结构、地理分布以及进化上与人类相近,开展其肠道微生物研究不仅有助于了解灵长类的生态、保护和进化,而且对深入了解肠道微生物在人类进化中所发挥的作用也具有重要的参考价值。本文总结了影响非人灵长类肠道微生物变化的因素,包括系统发育、觅食、栖息地破碎化、年龄和性别、圈养方式以及社群生活,并探讨了肠道微生物研究在非人灵长类生态、行为、保护以及适应性进化方面的应用。未来,非人灵长类肠道微生物研究将为灵长类生态、进化和人类健康的研究提供新的视角,为灵长类的保护提供新的理论基础和研究方法。     

哺乳动物肠道微生物与碳水化合物的互作及其影响

肠道微生物具有调节宿主营养、免疫以及能量代谢等生理功能。饮食是影响哺乳动物的肠道微生物的一个重要因素。碳水化合物是哺乳动物食物能量的主要来源,因此研究肠道微生物与碳水化合物的代谢之间的关系及其影响具有重要意义。基于近年相关研究,本文从碳水化合物对肠道微生物组成的影响、肠道微生物对碳水化合物的代谢机制以及碳水化合物发酵产物短链脂肪酸对宿主的影响3个方面进行了综述。研究表明,肠道微生物可用于发酵的碳水化合物类型主要是抗性淀粉和非淀粉多糖;不同类型的碳水化合物会导致肠道菌群发生适应性变化;复杂多糖发酵产生的短链脂肪酸在调节宿主能量平衡和免疫应答方面发挥了重要作用。总结近年来相关研究,可加深对肠道菌群对宿主碳水化合物代谢贡献的理解,为哺乳动物机体健康状况的营养调控策略提供参考。     

Environmental factors have a strong impact on the composition and diversity of the gut bacterial community of Chinese black honeybees

The Chinese black honeybees is an ecotype of the European honeybee that is formed by the natural hybridization of Apis mellifera mellifera and A. m. carnica. It is distributed in nature reserves in North China and has been an important breeding resource for disease resistance and other desirable traits. Compared to the areas outside of reserves, the nature reserves offer significant biodiversity benefits not only to the Chinese black honey bees but also to the other valuable plants and animals. In recent years there has been growing evidence that environmental factors including food choices play an important role in shaping the composition and activity of gut microbiota, which in turn can impact host health. In the previous studies on Chinse black honeybees, little attention has been paid to the diverse population of microbes in the gut that play a vital role in host health. In order to achieve a better understanding on the role of environmental factors in diversity and composition of gut microbiota of honey bees, in the present study, we analyzed the gut bacterial communities of Chinese black honeybees using terminal restriction fragment length polymorphism (T-RFLP). The results showed that the samples from the national nature reserves that are protected and managed so as to preserve and enrich their natural condition and resources for Chinese black honeybees had higher variety and richness of gut bacteria than that collected from unreserved regions that also harbor populations of Chinese black honeybees. The four terminal restriction fragments (T-RFs), 201, 223, 247 and 320 bp, were identified to be the dominant bacteria of Chinese black honeybees. Of which 247 and 320 bp had greater differences between bee groups sampled in different regions and therefore could be used as genetic markers to separate samples collected from the national nature reserves to samples collected from unreserved regions. The results clearly indicate that national nature reserve protects biological diversity and ecological and evolutionary processes which have had a significant influence on the diversity of gut bacteria of Chinese black honeybees. The ubiquity of gut symbiotic bacteria identified in Chinese black honeybee suggests that environmental factors could play an important role in diversity and composition of gut bacteria and warrant further investigation into the functional significance of these gut bacteria for the honeybee health.     

Gut microbiome dysbiosis and honeybee health

Since a few decades, apiculture is facing important economic losses worldwide with general major consequences in many areas of agriculture. A strong attention has been paid towards the phenomenon named Colony Collapse Disorder in which colonies suddenly disappear with no clear explanations. Honeybee colonies can be affected by abiotic factors, such as environmental pollution or insecticide applications for agricultural purposes. Also biotic stresses cause colony losses, including bacterial (e.g. Paenibacillus larvae) and fungal (e.g. Ascosphaera apis) pathogens, microsporidia (e.g. Nosema apis), parasites (i.e. Varroa destructor) and several viruses. In the light of recent research, intestinal dysbiosis, considered as the relative disproportion of the species within the native microbiota, has shown to affect human and animal health. In arthropods, alteration of the gut microbial climax community has been shown to be linked to health and fitness disequilibrium, like in the medfly Ceratitis capitata for which low mate competitiveness is determined by a gut microbial community imbalance. According to these observations, it is possible to hypothesize that dysbiosis may have a role in disease occurrence also in honeybees. Here we aim to discuss the current knowledge on dysbiosis in the honeybee and its relation with honeybee health by reviewing the investigations of the microbial diversity associated to honeybees and the recent experiments performed to control bee diseases by microbial symbionts. We conclude that, despite the importance of a good functionality of the associated microbiota in preserving insect health has been proved, the mechanisms involved in honeybee gut dysbiosis are still unknown. Accurate in vitro, in vivo and in field investigations are required under healthy, diseased and stressed conditions for the host.     

蜜蜂大脑的分区与功能

蜜蜂Apis mellifera L.是神经生物学研究的重要模式生物。尽管工蜂脑的体积不足1mm3,包含的神经元数量不到百万,但却拥有丰富的个体和社会行为,甚至还有学习、记忆、认知等高级行为。如此微小的大脑也是通过不同结构分区来实现其丰富复杂的行为。本文对蜜蜂大脑的精细解剖结构以及脑区功能研究进行了综述,为昆虫科学和神经生物学研究提供参考。     

Gut microbiome helps honeybee (Apis mellifera) resist the stress of toxic nectar plant (Bidens pilosa) exposure: Evidence for survival and immunity

Honeybee (Apis mellifera) ingestion of toxic nectar plants can threaten their health and survival. However, little is known about how to help honeybees mitigate the effects of toxic nectar plant poisoning. We exposed honeybees to different concentrations of Bidens pilosa flower extracts and found that B. pilosa exposure significantly reduced honeybee survival in a dose-dependent manner. By measuring changes in detoxification and antioxidant enzymes and the gut microbiome, we found that superoxide dismutase, glutathione-S-transferase and carboxylesterase activities were significantly activated with increasing concentrations of B. pilosa and that different concentrations of B. pilosa exposure changed the structure of the honeybee gut microbiome, causing a significant reduction in the abundance of Bartonella (p < 0.001) and an increase in Lactobacillus. Importantly, by using Germ-Free bees, we found that colonization by the gut microbes Bartonella apis and Apilactobacillus kunkeei (original classification as Lactobacillus kunkeei) significantly increased the resistance of honeybees to B. pilosa and significantly upregulated bee-associated immune genes. These results suggest that honeybee detoxification systems possess a level of resistance to the toxic nectar plant B. pilosa and that the gut microbes B. apis and A. kunkeei may augment resistance to B. pilosa stress by improving host immunity.     

Diversity and evolutionary patterns of bacterial gut associates of corbiculate bees

The animal gut is a habitat for diverse communities of microorganisms (microbiota). Honeybees and bumblebees have recently been shown to harbour a distinct and species poor microbiota, which may confer protection against parasites. Here, we investigate diversity, host specificity and transmission mode of two of the most common, yet poorly known, gut bacteria of honeybees and bumblebees: Snodgrassella alvi (Betaproteobacteria) and Gilliamella apicola (Gammaproteobacteria). We analysed 16S rRNA gene sequences of these bacteria from diverse bee host species across most of the honeybee and bumblebee phylogenetic diversity from North America, Europe and Asia. These focal bacteria were present in 92% of bumblebee species and all honeybee species but were found to be absent in the two related corbiculate bee tribes, the stingless bees (Meliponini) and orchid bees (Euglossini). Both Snodgrassella alvi and Gilliamella apicola phylogenies show significant topological congruence with the phylogeny of their bee hosts, albeit with a considerable degree of putative host switches. Furthermore, we found that phylogenetic distances between Gilliamella apicola samples correlated with the geographical distance between sampling locations. This tentatively suggests that the environmental transmission rate, as set by geographical distance, affects the distribution of G. apicola infections. We show experimentally that both bacterial taxa can be vertically transmitted from the mother colony to daughter queens, and social contact with nest mates after emergence from the pupa greatly facilitates this transmission. Therefore, sociality may play an important role in vertical transmission and opens up the potential for co‐evolution or at least a close association of gut bacteria with their hosts.     

Geographically widespread honeybee‐gut symbiont subgroups show locally distinct antibiotic‐resistant patterns

How long‐term antibiotic treatment affects host bacterial associations is still largely unknown. The honeybee‐gut microbiota has a simple composition, so we used this gut community to investigate how long‐term antibiotic treatment affects host‐associated microbiota. We investigated the phylogenetic relatedness, genomic content (GC percentage, genome size, number of genes and CRISPR) and antibiotic‐resistant genes (ARG) for strains from two abundant members of the honeybee core gut microbiota (Gilliamella apicola and Snodgrassella alvi). Domesticated honeybees are subjected to geographically different management policies, so we used two research apiaries, representing different antibiotic treatment regimens in their apiculture: low antibiotic usage (Norway) and high antibiotic usage (Arizona, USA). We applied whole‐genome shotgun sequencing on 48 G. apicola and 22 S. alvi. We identified three predominating subgroups of G. apicola in honeybees from both Norway and Arizona. For G. apicola, genetic content substantially varied between subgroups and distance similarity calculations showed similarity discrepancy between subgroups. Functional differences between subgroups, such as pectin‐degrading enzymes (G. apicola), were also identified. In addition, we identified horizontal gene transfer (HGT) of transposon (Tn10)‐associated tetracycline resistance (Tet B) across the G. apicola subgroups in the Arizonan honeybees, using interspace polymorphisms in the Tet B determinant. Our results support that honeybee‐gut symbiont subgroups can resist long‐term antibiotic treatment and maintain functionality through acquisition of geographically distinct antibiotic‐resistant genes by HGT.     

A selective sweep in a microsporidian parasite Nosema‐tolerant honeybee population,Apis mellifera

Nosema is a microsporidian parasite of the honeybee, which infects the epithelial cells of the gut. In Denmark, honeybee colonies have been selectively bred for the absence of Nosema over decades, resulting in a breeding line that is tolerant toward Nosema infections. As the tolerance toward the Nosema infection is a result of artificial selection, we screened chromosome 14 for a selective sweep with microsatellite markers, where a major quantitative trait locus (QTL) had been identified to be involved in the reduction in Nosema spores in the honeybees. By comparing the genetic variability of 10 colonies of the selected honeybee strain with a population sample from 22 unselected colonies, a selective sweep was revealed within the previously identified QTL region. The genetic variability of the swept loci was not only reduced in relation to the flanking markers on chromosome 14 within the selected strain but also significantly reduced compared with the same region in the unselected honeybees. This confirmed the results of the previous QTL mapping for reduced Nosema infections. The success of the selective breeding may have driven the selective sweep found in our study.     

蜜蜂转基因研究进展及研究途径探讨

转基因动物的科研价值和商业价值促进了转基因技术的不断发展和在各个领域的深入应用。蜜蜂是有着悠久饲养历史的经济昆虫和在基础理论研究领域有重大应用价值的模式动物,但其转基因研究却相对落后。雌性蜂的级型分化和工蜂清洁巢房行为增加了蜜蜂转基因的难度,精子介导转基因配套以人工授精技术及蜜蜂卵或幼虫的转基因操作与蜜蜂人工孵育技术结合是目前蜜蜂转基因的较好途径。文章综述蜜蜂转基因的研究进展,并讨论蜜蜂转基因所面临的特殊性及其研究途径。     

引入西方蜜蜂对中蜂的危害及生态影响

作者阐述自1896年中国引进西方蜜蜂Apis melliferaL.的优良品种如意大利蜂Apis mellifera ligusticaSpinola和喀尼阿兰蜂Apis mellifera Carnica Pollmann以来,使当地的东方蜜蜂Apis cerana F.受到严重危害,其分布区域缩小75%以上,种群数量减少80%以上。使山林植物授粉总量减少,导致植物多样性减少。文中提出: 建立自然保护区保存本地蜜蜂遗传特性,和采用基因转移等技术,培育具有西方蜜蜂优良生产性能的中蜂新品种,逐步恢复中蜂的种群数量。