Bacteriome

细菌组
  • 文章类型: Journal Article
    土壤微生物组科学,快速发展,主要集中在田间作物土壤上。然而,了解花园土壤微生物对于提高花园环境中粮食生产的可持续性至关重要。这项研究旨在揭示达卡地区城市(达卡北部和达卡南部城市公司)和城市周边地区(加济布尔市公司)的屋顶花园土壤(RGS)和地表花园土壤(SGS)的细菌多样性和组成。孟加拉国。我们分析了11个样本,包括来自11个单独花园的6个RGS和5个SGS样本,使用基于16SrRNA(V3-V4区域)基因的扩增子测序。共有977个操作分类单位(OTU),包括RGS和SGS样品中的270和707,分别,已确定。观察到的OTU由21门代表,45班,84个订单,173个家庭,和293属细菌。Alpha多样性指数显示SGS样本中细菌多样性显著较高(p=0.01),而β多样性分析表明RGS和SGS样品之间有不同的细菌组组成(p=0.028,PERMANOVA)。尽管样本类别之间的分类差异很大,也有相当多的共享细菌类群的存在。在门一级,芽孢杆菌(61.14%),Pseudomonadota(23.42%),放线菌(6.33%),和拟杆菌(3.32%)是两种类型的花园土壤样品中的主要细菌门(占总丰度的94.0%)。在确定的属中,RGS中的芽孢杆菌(69.73%)和短芽孢杆菌(18.81%)和芽孢杆菌(19.22%),甲基phaga(19.21%),不动杆菌(6.27%),棒状杆菌(5.06%),伯克霍尔德菌(4.78%),SGS中副球菌(3.98%)和溶杆菌(2.07%)是主要的细菌属。重要的是,我们检测到52.90%的属在RGS和SGS土壤样品之间共享。我们的数据揭示了来自屋顶和地面花园的土壤样品中具有益生菌潜力的独特和共享的细菌。进一步的研究应探索共享细菌类群在花园土壤中的功能作用,以及城市环境因素如何影响微生物组组成,以优化土壤健康和可持续粮食生产。
    Soil microbiome science, rapidly evolving, predominantly focuses on field crop soils. However, understanding garden soil microbiomes is essential for enhancing food production sustainability in garden environments. This study aimed to unveil the bacteriome diversity and composition in rooftop garden soils (RGS) and surface garden soils (SGS) across urban (Dhaka North and Dhaka South City Corporations) and peri-urban (Gazipur City Corporation) areas of Dhaka Division, Bangladesh. We analyzed 11 samples, including six RGS and five SGS samples from 11 individual gardens using 16S rRNA (V3-V4 region) gene-based amplicon sequencing. A total of 977 operational taxonomic units (OTUs), including 270 and 707 in RGS and SGS samples, respectively, were identified. The observed OTUs were represented by 21 phyla, 45 classes, 84 orders, 173 families, and 293 genera of bacteria. Alpha diversity indices revealed significantly higher bacterial diversity in SGS samples (p = 0.01), while beta diversity analyses indicated distinct bacteriome compositions between RGS and SGS samples (p = 0.028, PERMANOVA). Despite substantial taxonomic variability between sample categories, there was also a considerable presence of shared bacterial taxa. At the phylum level, Bacilliota (61.14%), Pseudomonadota (23.42%), Actinobacteria (6.33%), and Bacteroidota (3.32%) were the predominant bacterial phyla (comprising > 94.0% of the total abundances) in both types of garden soil samples. Of the identified genera, Bacillus (69.73%) and Brevibacillus (18.81%) in RGS and Bacillus (19.22%), Methylophaga (19.21%), Acinetobacter (6.27%), Corynebacterium (5.06%), Burkholderia (4.78%), Paracoccus (3.98%) and Lysobacter (2.07%) in SGS were the major bacterial genera. Importantly, we detected that 52.90% of genera were shared between RGS and SGS soil samples. Our data reveal unique and shared bacteriomes with probiotic potential in soil samples from both rooftop and surface gardens. Further studies should explore the functional roles of shared bacterial taxa in garden soils and how urban environmental factors affect microbiome composition to optimize soil health and sustainable food production.
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  • 文章类型: Journal Article
    墨西哥中太平洋(MCP)地区具有不连续的珊瑚生态系统,具有不同的保护和人为干扰。表征与海胆弓形虫相关的细菌组合及其与环境变量的关系将有助于理解该物种的生理和生态学。我们连续两年在夏季和冬季从MCP的六个地点的珊瑚生态系统中收集海胆。空间尺度代表了T.roseus细菌组最重要的变异,特别是因为IslaIsabel国家公园(PNII)。同样,与生境结构变量相关的空间差异,主要是海绵和活珊瑚覆盖。与其他地点相比,PNII表现出高度多样化的细菌组合,以与疾病和环境压力相关的家庭为特征(腐植科,Flammeovirgaceae,和黄杆菌科)。其余五个地点呈现恒定的时空模式,其中弯曲杆菌科和螺旋杆菌科的优势是玫瑰T.holobiont的关键。然而,某些细菌家族的优势,如肠杆菌科,在分析的第二年,表明PuntoB和IslaseislototesdeBahíaChamelaSanctuary暴露于污水污染。总的来说,我们的结果提高了对特定时间和空间中宿主相关细菌组合及其与环境条件关系的理解。
    The Mexican Central Pacific (MCP) region has discontinuous coral ecosystems with different protection and anthropogenic disturbance. Characterizing the bacterial assemblage associated with the sea urchin Toxopneustes roseus and its relationship with environmental variables will contribute to understanding the species\' physiology and ecology. We collected sea urchins from coral ecosystems at six sites in the MCP during the summer and winter for two consecutive years. The spatial scale represented the most important variation in the T. roseus bacteriome, particularly because of Isla Isabel National Park (PNII). Likewise, spatial differences correlated with habitat structure variables, mainly the sponge and live coral cover. The PNII exhibited highly diverse bacterial assemblages compared to other sites, characterized by families associated with diseases and environmental stress (Saprospiraceae, Flammeovirgaceae, and Xanthobacteraceae). The remaining five sites presented a constant spatiotemporal pattern, where the predominance of the Campylobacteraceae and Helicobacteraceae families was key to T. roseus\' holobiont. However, the dominance of certain bacterial families, such as Enterobacteriaceae, in the second analyzed year suggests that Punto B and Islas e islotes de Bahía Chamela Sanctuary were exposed to sewage contamination. Overall, our results improve the understanding of host-associated bacterial assemblages in specific time and space and their relationship with the environmental condition.
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  • 文章类型: Journal Article
    目前支气管扩张的治疗方法,这是由气道的病理性扩张定义的,仅限于症状缓解和减少恶化。这种情况在世界范围内变得越来越普遍。由于该疾病的病理生理学尚未完全了解,开发新的治疗方法至关重要。慢性感染的相互作用,炎症,粘膜纤毛清除受损,导致结构改变和新感染的出现,最有可能导致支气管扩张的进展。除了治疗囊性纤维化引起的支气管扩张,没有批准的治疗方法。了解微生物在这种疾病中的参与至关重要,微生物组被定义为环境中所有细菌的集体遗传物质。在临床实践中,已经使用培养物研究了肺中的细菌;然而,近年来,研究人员使用下一代测序方法,如16SrRNA测序。虽然支气管扩张中的微生物组尚未得到完全研究,我们所知道的表明嗜血杆菌,假单胞菌和链球菌在肺部细菌生态系统中占主导地位,它们表现出显著的个体内稳定性和个体间异质性。假单胞菌和嗜血杆菌为主的微生物群与更严重的疾病和频繁的恶化有关,然而,需要更多的研究来充分了解微生物组在支气管扩张演变中的作用。这篇综述讨论了有关肺部微生物群及其与支气管扩张的关系的最新发现。
    The present treatments for bronchiectasis, which is defined by pathological dilatation of the airways, are confined to symptom relief and minimizing exacerbations. The condition is becoming more common worldwide. Since the disease\'s pathophysiology is not entirely well understood, developing novel treatments is critically important. The interplay of chronic infection, inflammation, and compromised mucociliary clearance, which results in structural alterations and the emergence of new infection, is most likely responsible for the progression of bronchiectasis. Other than treating bronchiectasis caused by cystic fibrosis, there are no approved treatments. Understanding the involvement of the microbiome in this disease is crucial, the microbiome is defined as the collective genetic material of all bacteria in an environment. In clinical practice, bacteria in the lungs have been studied using cultures; however, in recent years, researchers use next-generation sequencing methods, such as 16S rRNA sequencing. Although the microbiome in bronchiectasis has not been entirely investigated, what is known about it suggests that Haemophilus, Pseudomonas and Streptococcus dominate the lung bacterial ecosystems, they present significant intraindividual stability and interindividual heterogeneity. Pseudomonas and Haemophilus-dominated microbiomes have been linked to more severe diseases and frequent exacerbations, however additional research is required to fully comprehend the role of microbiome in the evolution of bronchiectasis. This review discusses recent findings on the lung microbiota and its association with bronchiectasis.
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  • 文章类型: Journal Article
    数十年来,研究人员一直未能解开多种症状的海湾战争疾病(GWI)病理并找到有效的治疗方法。在小鼠模型中研究病因的慢性症状持续性和局限性与人类的病因显着不同,这对药物发现和寻找有效的治疗方案构成了挑战。GWI暴露在研究队列中存在显着差异,以上情况使得很难重建与GWI症状病理学非常相似的模型。我们已经使用双重植入策略来重建人类免疫系统以及人类微生物组转移,以创建GWI的人源化小鼠模型。采用全基因组鸟枪测序和血液免疫细胞因子酶联免疫吸附试验(ELISA),我们表明,我们用海湾战争(GW)化学物质处理的双人化小鼠显示出显着改变的肠道微生物组,与GWI的退伍军人队列中报道的相似。结果还显示了相似的细胞因子谱,如IL-1β水平升高,双人源化模型中的IL-6和TNF-R-1,正如以前在人类队列中发现的那样。Further,我们利用一种新型GWI退伍军人粪便微生物群转移方法创建了第二种替代模型,该模型与GWI退伍军人的微生物组和免疫系统相关病理非常相似.在人源化小鼠中的GWI退伍军人微生物群移植显示了人类微生物组重建和全身性炎症病理,正如白细胞介素1β的增加所反映的那样,6,8(IL-1β,IL-6,IL-8),肿瘤坏死因子受体1(TNF-R-1),和内毒素血症。总之,虽然是初步的,我们报道了一种新的体内模型,该模型具有人类微生物组重建和移植的人类免疫表型,可能有助于更好地了解GWI中的肠道-免疫相互作用.
    Unraveling the multisymptomatic Gulf War Illness (GWI) pathology and finding an effective cure have eluded researchers for decades. The chronic symptom persistence and limitations for studying the etiologies in mouse models that differ significantly from those in humans pose challenges for drug discovery and finding effective therapeutic regimens. The GWI exposome differs significantly in the study cohorts, and the above makes it difficult to recreate a model closely resembling the GWI symptom pathology. We have used a double engraftment strategy for reconstituting a human immune system coupled with human microbiome transfer to create a humanized-mouse model for GWI. Using whole-genome shotgun sequencing and blood immune cytokine enzyme linked immunosorbent assay (ELISA), we show that our double humanized mice treated with Gulf War (GW) chemicals show significantly altered gut microbiomes, similar to those reported in a Veteran cohort of GWI. The results also showed similar cytokine profiles, such as increased levels of IL-1β, IL-6, and TNF R-1, in the double humanized model, as found previously in a human cohort. Further, a novel GWI Veteran fecal microbiota transfer was used to create a second alternative model that closely resembled the microbiome and immune-system-associated pathology of a GWI Veteran. A GWI Veteran microbiota transplant in humanized mice showed a human microbiome reconstitution and a systemic inflammatory pathology, as reflected by increases in interleukins 1β, 6, 8 (IL-1β, IL-6, IL-8), tumor necrosis factor receptor 1 (TNF R-1), and endotoxemia. In conclusion, though preliminary, we report a novel in vivo model with a human microbiome reconstitution and an engrafted human immune phenotype that may help to better understand gut-immune interactions in GWI.
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  • 文章类型: Journal Article
    许多昆虫以植物汁液等营养挑战性饮食为食,叶子,或木材与细菌共生体有古老的联系,这些细菌共生体补充限制性营养或产生消化或解毒酶。然而,分布,函数,以及利用其他植物组织或依靠饮食的昆虫中微生物共生体的进化动力学仍然知之甚少。这里,我们研究了软翼花甲虫(Melyridae,Dashtinae)在幼虫和成虫阶段之间从食性或食肉性过渡到古生性(花粉摄食)。重建Dashtinae系统发育中共生体的分布揭示了长期的共同进化,源于单一的收购事件,随后的宿主-共生体共同多元化,但也有几个独立的共生体损失。对20种不同共生体基因组的分析表明,它们的基因组受到严重侵蚀。然而,普遍保留的莽草酸途径表明,对其宿主的核心代谢贡献是为角质层硬化和黑化提供酪氨酸。尽管共生体菌株的基因含量和顺序高度相似,合成额外必需氨基酸和维生素以及回收尿素的能力保留在一些但不是所有的共生体中,表明寄主谱系之间的生态差异。这篇关于昆虫与食腐或食肉幼虫和花粉摄食成虫的酪氨酸共生体的报道,扩大了我们对酪氨酸补充剂的理解,这是一种重要的共生体提供的益处,适用于具有多种摄食生态的广泛昆虫。
    Many insects feeding on nutritionally challenging diets like plant sap, leaves, or wood engage in ancient associations with bacterial symbionts that supplement limiting nutrients or produce digestive or detoxifying enzymes. However, the distribution, function, and evolutionary dynamics of microbial symbionts in insects exploiting other plant tissues or relying on a predacious diet remain poorly understood. Here, we investigated the evolutionary history and function of the intracellular gamma-proteobacterial symbiont \"Candidatus Dasytiphilus stammeri\" in soft-winged flower beetles (Coleoptera, Melyridae, Dasytinae) that transition from saprophagy or carnivory to palynivory (pollen-feeding) between larval and adult stage. Reconstructing the distribution of the symbiont within the Dasytinae phylogeny unraveled not only a long-term coevolution, originating from a single acquisition event with subsequent host-symbiont codiversification, but also several independent symbiont losses. The analysis of 20 different symbiont genomes revealed that their genomes are severely eroded. However, the universally retained shikimate pathway indicates that the core metabolic contribution to their hosts is the provisioning of tyrosine for cuticle sclerotization and melanization. Despite the high degree of similarity in gene content and order across symbiont strains, the capacity to synthesize additional essential amino acids and vitamins and to recycle urea is retained in some but not all symbionts, suggesting ecological differences among host lineages. This report of tyrosine-provisioning symbionts in insects with saprophagous or carnivorous larvae and pollen-feeding adults expands our understanding of tyrosine supplementation as an important symbiont-provided benefit across a broad range of insects with diverse feeding ecologies.
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  • 文章类型: Journal Article
    多发性硬化症(MS)源于宿主遗传因素和环境成分之间复杂的相互作用,随着肠道微生物群成为一个关键的调查领域。在目前的研究中,我们使用离子洪流测序来深入研究MS(pwMS)患者的细菌组(细菌微生物群)和分枝杆菌组(真菌微生物群),并将其与健康对照(HC)进行比较。通过主要协调,多样性,和丰度分析,以及聚类和跨界微生物相关性评估,我们发现pwMS和HC之间的微生物谱存在显着差异。在pwMS中观察到真菌Torulaspora和细菌类肠杆菌科的水平升高,而有益的细菌分类群,如普雷沃拉达科和Dialister,减少了。值得注意的是,聚类分析显示,74%的参与者的细菌组和分枝杆菌组数据存在重叠模式,在pwMS改变的微生物群中明显具有减弱的跨王国相互作用。我们的发现强调了MS中细菌和真菌微生物群的菌群失调,以生物多样性和成分的变化为特征。此外,真菌-细菌相互作用的独特疾病相关模式表明真菌,除了细菌,有助于MS的发病机制。总的来说,我们的研究揭示了MS背后复杂的微生物动力学,为进一步研究MS管理中肠道微生物群的潜在治疗靶向奠定了基础。
    Multiple sclerosis (MS) arises from a complex interplay between host genetic factors and environmental components, with the gut microbiota emerging as a key area of investigation. In the current study, we used ion torrent sequencing to delve into the bacteriome (bacterial microbiota) and mycobiome (fungal microbiota) of people with MS (pwMS), and compared them to healthy controls (HC). Through principal coordinate, diversity, and abundance analyses, as well as clustering and cross-kingdom microbial correlation assessments, we uncovered significant differences in the microbial profiles between pwMS and HC. Elevated levels of the fungus Torulaspora and the bacterial family Enterobacteriaceae were observed in pwMS, whereas beneficial bacterial taxa, such as Prevotelladaceae and Dialister, were reduced. Notably, clustering analysis revealed overlapping patterns in the bacteriome and mycobiome data for 74% of the participants, with weakened cross-kingdom interactions evident in the altered microbiota of pwMS. Our findings highlight the dysbiosis of both bacterial and fungal microbiota in MS, characterized by shifts in biodiversity and composition. Furthermore, the distinct disease-associated pattern of fungi-bacteria interactions suggests that fungi, in addition to bacteria, contribute to the pathogenesis of MS. Overall, our study sheds light on the intricate microbial dynamics underlying MS, paving the way for further investigation into the potential therapeutic targeting of the gut microbiota in MS management.
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  • 文章类型: Journal Article
    普通的臭虫,Cimexlectularius,是只以血液为食的半翅目昆虫,谁的叮咬会导致公共卫生问题。由于全球化和对杀虫剂的抗药性,近几十年来,这种害虫经历了重大的全球复苏。血液是一种不均衡的饮食,缺乏足够的B族维生素。像所有严格的食血节肢动物一样,臭虫拥有提供B族维生素的营养共生体。在C.lectularius,这种营养共生体是细胞内细菌Wolbachia(wCle)。它位于特定的共生器官中,细菌体,以及卵巢。wCle的实验性消耗已被证明会导致若虫发育更长和繁殖力更低。这些表型是通过补充B族维生素来拯救的。了解wCle与臭虫之间的相互作用可能有助于开发新的害虫防治方法,以破坏这种共生相互作用。因此,这项工作的目的是准确量化宿主生命周期中wCle的密度,并描述细菌组中潜在的相关形态变化。我们还试图确定性的影响,喂养状态,和衰老对细菌种群动态的影响。我们表明,在臭虫发育过程中,wCle的相对数量不断增加,而细菌组的相对大小保持稳定。我们还表明,成年女性比男性拥有更多的wCle,并且随着年龄的增长,成年女性的wCle相对数量略有下降。除了每周喂食的雄性。这些结果在臭虫生态学的背景下进行了讨论,将有助于确定臭虫生命周期中共生相互作用的关键点。
    The common bed bug, Cimex lectularius, is a hemipteran insect that feeds only on blood, and whose bites cause public health issues. Due to globalization and resistance to insecticides, this pest has undergone a significant and global resurgence in recent decades. Blood is an unbalanced diet, lacking notably sufficient B vitamins. Like all strict hematophagous arthropods, bed bugs host a nutritional symbiont supplying B vitamins. In C. lectularius, this nutritional symbiont is the intracellular bacterium Wolbachia (wCle). It is located in specific symbiotic organs, the bacteriomes, as well as in ovaries. Experimental depletion of wCle has been shown to result in longer nymphal development and lower fecundity. These phenotypes were rescued by B vitamin supplementation. Understanding the interaction between wCle and the bed bug may help to develop new pest control methods targeting the disruption of this symbiotic interaction. The objective of this work was thus to quantify accurately the density of wCle over the life cycle of the host and to describe potential associated morphological changes in the bacteriome. We also sought to determine the impact of sex, feeding status, and aging on the bacterial population dynamics. We showed that the relative quantity of wCle continuously increases during bed bug development, while the relative size of the bacteriome remains stable. We also showed that adult females harbor more wCle than males and that wCle relative quantity decreases slightly in adults with age, except in weekly-fed males. These results are discussed in the context of bed bug ecology and will help to define critical points of the symbiotic interaction during the bed bug life cycle.
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  • 文章类型: Journal Article
    Kuflu奶酪,一种流行的传统土耳其模具成熟奶酪,其特点是半坚硬的质地和蓝绿色。阐明从生乳生产的Kuflu奶酪的微生物群以标准化和维持其感官特性非常重要。这项研究旨在检查细菌,酵母,和Kuflu奶酪中的丝状霉菌群落,使用基于16S和ITS2区域的高通量扩增子测序。乳球菌,链球菌,葡萄球菌是最主要的细菌属,而在一些Kuflu奶酪样品中发现双歧杆菌属非常高。青霉属在丝状霉菌生物群中占主导地位,而相对丰度最高的酵母则被检测为脱巴酵母,Pichia,还有念珠菌.Virgibacillus和Paraliobacillus,以前没有报道过霉菌成熟的奶酪,在一些Kuflu奶酪样品中检测到高相对丰度。没有一个属包括沙门氏菌等重要的食物病原体,弯曲杆菌,在样品中检测到李斯特菌。这是用宏基因组方法评估Kuflu奶酪的微生物群的第一个实验。这项研究为评估Kuflu奶酪提供了机会,以前检查过真菌成分,在病原和有益细菌方面。
    Kuflu cheese, a popular variety of traditional Turkish mold-ripened cheeses, is characterized by its semi-hard texture and blue-green color. It is important to elucidate the microbiota of Kuflu cheese produced from raw milk to standardize and sustain its sensory properties. This study aimed to examine the bacteria, yeasts, and filamentous mold communities in Kuflu cheese using high-throughput amplicon sequencing based on 16S and ITS2 regions. Lactococcus, Streptococcus, and Staphylococcus were the most dominant bacterial genera while Bifidobacterium genus was found to be remarkably high in some Kuflu cheese samples. Penicillium genus dominated the filamentous mold biota while the yeasts with the highest relative abundances were detected as Debaryomyces, Pichia, and Candida. The genera Virgibacillus and Paraliobacillus, which were not previously reported for mold-ripened cheeses, were detected at high relative abundances in some Kuflu cheese samples. None of the genera that include important food pathogens like Salmonella, Campylobacter, Listeria were detected in the samples. This is the first experiment in which the microbiota of Kuflu cheeses were evaluated with a metagenomic approach. This study provided an opportunity to evaluate Kuflu cheese, which was previously examined for fungal composition, in terms of both pathogenic and beneficial bacteria.
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  • 文章类型: Journal Article
    背景:肠道微生物群对人类高海拔适应的贡献仍未得到充分理解。
    方法:在这里,使用深全宏基因组鸟枪测序对生活在海平面和高海拔地区的健康个体进行了肠道微生物群的比较分析,探讨高原居民肠道菌群的适应机制。
    结果:结果显示,与海平面个体相比,高海拔个体的肠道细菌组表现出更大的样本内多样性和细菌组成和功能谱的显着变化。表明与高海拔环境相关的独特细菌的潜在选择。菌株水平的调查显示,高海拔人群中Collinsellaaerorofaciens和Akkermansiamuciniphila的富集。还研究了肠道病毒和肠道真菌的特征。与海平面相比,高海拔受试者的肠道病毒表现出更大的多样性,随着病毒操作分类单位(vOTU)和独特的注释基因数量的增加。最后,相关性分析显示,42种细菌和375种vOTUs之间有819个显著相关性,而细菌和真菌之间或真菌和病毒之间没有观察到显著的相关性。
    结论:这些发现极大地促进了对人类肠道菌群高海拔地理适应机制的理解。
    BACKGROUND: The contribution of gut microbiota to human high-altitude adaptation remains inadequately understood.
    METHODS: Here a comparative analysis of gut microbiota was conducted between healthy individuals living at sea level and high altitude using deep whole-metagenome shotgun sequencing, to investigate the adaptive mechanisms of gut microbiota in plateau inhabitants.
    RESULTS: The results showed the gut bacteriomes in high-altitude individuals exhibited greater within-sample diversity and significant alterations in both bacterial compositional and functional profiles when compared to those of sea-level individuals, indicating the potential selection of unique bacteria associated with high-altitude environments. The strain-level investigation revealed enrichment of Collinsella aerofaciens and Akkermansia muciniphila in high-altitude populations. The characteristics of gut virome and gut mycobiome were also investigated. Compared to sea-level subjects, high-altitude subjects exhibited a greater diversity in their gut virome, with an increased number of viral operational taxonomic units (vOTUs) and unique annotated genes. Finally, correlation analyses revealed 819 significant correlations between 42 bacterial species and 375 vOTUs, while no significant correlations were observed between bacteria and fungi or between fungi and viruses.
    CONCLUSIONS: The findings have significantly contributed to an enhanced comprehension of the mechanisms underlying the high-altitude geographic adaptation of the human gut microbiota.
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  • 文章类型: Journal Article
    目的:研究患有原发性牙髓感染(PEI)和根尖周炎(AP)的牙齿中存在的细菌组,并定量和定性地确定使用2.5%次氯酸钠NAOCl的化学机械制剂(CMP)对使用IlluminaMiSeq平台在PEI和AP中发现的细菌组的影响。
    方法:对18例患者的36个配对样本进行了成功测序和分析。在两个采样时间收集样品:在使用2.5%NaOCl的CMP之前(s1)和之后(s2)。从s1和s2样品中提取DNA并使用定量PCR(qPCR)定量。使用IlluminaMiSeq平台对所有36个样品进行测序。用DADA2流水线处理原始V3-V4扩增子测序数据以产生扩增子序列变体(ASV)。代表丰度(Chao1)和多样性和均匀度(Shannon,辛普森)进行了计算。配对样品Wilcoxon检验用于比较s1和s2之间的α多样性指标和qPCR计数。应用PERMANOVA方法(具有999个排列)来比较样品类型(s1对s2)之间和患者ID之间的群落组成。ALDEx2(用于高通量测序数据的ANOVA样差异表达工具)研究s1和s2之间的差异丰富分类单元。使用配对样品Wilcoxon检验来比较s1和s2之间的α多样性指标和qPCR计数。
    结果:与s2相比,s1中的qPCR计数明显更高(p=.0007)。Chao1指数表明α多样性没有差异(p<.7019);而Shannon(p=.0056)和Simpson(p=.02685)指数在s2中显示出较高的值。使用Adonis2的PERMANOVA检验显示样本时间对群落组成有显著影响(R2=.0630,p=.012)。患者ID对社区组成也显示出显着影响(R2=.6961,p=.001)。在属一级,Dialister,小杆菌属,普雷沃氏菌,和Olsenella在s1差异富集,而放线菌,窄食单胞菌_未分类,肠球菌_未分类,和未分类的放线菌在s2中差异富集。
    结论:PEI伴AP的牙齿中存在的细菌组复杂多样。使用2.5%NaOCl的CMP对带有AP的PEI中存在的细菌组显示出较高的定量和定性消毒剂影响。
    OBJECTIVE: To investigate the bacteriome present in teeth with primary endodontic infection (PEI) and apical periodontitis (AP) and to determine quantitatively and qualitatively the impact of chemomechanical preparation (CMP) using 2.5% sodium hypochlorite NAOCl on the bacteriome found in PEI with AP using the Illumina MiSeq platform.
    METHODS: Thirty-six paired samples from 18 patients were successfully sequenced and analysed. Samples were collected at two sampling times: before (s1) and after (s2) CMP using 2.5% NaOCl. The DNA was extracted from s1 and s2 samples and quantified using quantitative PCR (qPCR). All 36 samples were sequenced using the Illumina MiSeq platform. Raw V3-V4 amplicon sequencing data were processed with the DADA2 pipeline to generate amplicon sequence variants (ASVs). Alpha diversity metrics representing abundance (Chao1) and diversity and evenness (Shannon, Simpson) were computed. The paired-sample Wilcoxon\'s test was used to compare alpha diversity metrics and qPCR counts between s1 and s2. The PERMANOVA method (with 999 permutations) was applied to compare community composition between sample types (s1 versus s2) and between patient IDs. ALDEx2 (ANOVA-like differential expression tool for high-throughput sequencing data) to investigate differentially abundant taxa between s1 and s2. A paired-sample Wilcoxon\'s test was used to compare alpha diversity metrics and qPCR counts between s1 and s2.
    RESULTS: The qPCR counts were significantly higher in s1 compared to s2 (p = .0007). The Chao1 index indicated no difference in alpha diversity (p < .7019); whereas Shannon (p = .0056) and Simpson (p = .02685) indexes showed higher values in s2. The PERMANOVA test using Adonis2 showed a significant effect of sample time on community composition (R2 = .0630, p = .012). Patient ID also showed a significant effect on community composition (R2 = .6961, p = .001). At the genus level, Dialister, Mogibacterium, Prevotella, and Olsenella were differentially enriched at s1, while Actinomyces, Stenotrophomonas_unclassified, Enterococcus_unclassified, and Actinomyces_unclassified were differentially enriched in s2.
    CONCLUSIONS: The bacteriome present in teeth with PEI with AP is complex and diverse. CMP using 2.5% NaOCl showed a high quantitatively and qualitatively disinfectant impact on the bacteriome present in PEI with AP.
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