16S amplicon sequencing

16S 扩增子测序
  • 文章类型: Journal Article
    目的:口腔健康被认为是衰老的重要指标。口腔微生物群与口腔健康高度相关,而老年人口口腔微生物组的变化和与衰老相关的特征性微生物仍不清楚。
    方法:在本研究中,纳入130名老年受试者,按年龄分为3组:I期组(65≤70岁),II期组(70岁≤75岁),和III期组(75≤80岁)。使用IlluminaMiSeq平台分析其生理指标,并通过高通量测序确定口腔微生物组。
    结果:随着年龄的增长,空腹血糖水平,收缩压和单核细胞显着增加。各组间口腔微生物组整体结构无显著差异。在使用Metastats和Spearman\的相关分析时,特定细菌被鉴定为潜在的年龄或健康指数相关细菌属,包括梭杆菌属,Parvimonas,卟啉单胞菌,氨基杆菌,Collinsella,梭菌和不动杆菌。
    结论:我们的研究表明,老年人口唾液菌群的组成结构相对稳定,而特定细菌与年龄和健康状况相关。经过进一步探索,有望作为老年人的健康指标。
    OBJECTIVE: The health of oral cavity is considered as an important indicator of aging. Oral microbiota is highly associated with the oral health, while the variation of oral microbiome in elderly population and characteristic microbes associated with aging remain unclear.
    METHODS: In this study, 130 elderly subjects were recruited and divided into 3 groups according to their age: Stage I group (65 ≤ years < 70), Stage II group (70 ≤ years < 75), and Stage III group (75 ≤ years < 80). Their physiological indices were analyzed with using Illumina MiSeq platform and the oral microbiome was determined by high-throughput sequencing.
    RESULTS: Along with aging, the level of fasting blood glucose, systolic pressure and monocytes are significantly increased. No significant difference was detected on the whole structure of the oral microbiome among groups. While using Metastats and Spearman\'s correlation analysis, specific bacteria were identified as potential age- or health index-related bacterial genera including Fusobacterium, Parvimonas, Porphyromonas, Aminobacter, Collinsella, Clostridium and Acinetobacter.
    CONCLUSIONS: Our study revealed that the composition structure of salivary microbiota in elderly population was relatively stable while specific bacteria were correlated with age and health status, which is promising to be served as health indicators of the elderly after further exploration.
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  • 文章类型: Journal Article
    背景:龋齿的状况与口腔微生物组的变化密切相关。在这项研究中,我们比较了儿童严重早期龋齿(S-ECC)患儿在全身麻醉和门诊治疗前后牙菌斑微生物组的多样性和结构.
    方法:选择40名在全身麻醉(C1)或门诊(C2)下完成全口牙科治疗的3至5岁S-ECC患儿,每组20人。记录患儿的基本信息和口腔健康状况,以及治疗前牙菌斑的微生物群落结构和多样性(C1,C2),治疗后的第二天(C2_0D),治疗后7天(C1_7D,C2_7D),治疗后1个月(C1_1M,C2_1M),和治疗后3个月(C1_3M,C2_3M)通过16SrRNA高通量测序技术进行分析。
    结果:(1)α多样性检验表明,多重组菌群丰富度在治疗后明显大于治疗前(P<0.05),其余α多样性指数在两组间差异不显著(P>0.05)。β多样性分析表明,C1_7D组和C2_3M组的植物区系结构与各组内其他时间点的差异有统计学意义(P<0.05)。(2)治疗前和治疗后两组均存在核心菌群。根据两组儿童的龋齿状况,它们的植物丰度比例可能会发生变化。单约会组和多约会组治疗后7天,Leptotrichia丰度显着降低(P<0.05)。C1_1M和C1_3M组比C1和C1_7D组显著丰富(P<0.05)。链球菌,C1_7D组的嗜血杆菌和副流感嗜血杆菌明显高于其他组(P<0.05)。
    结论:全身麻醉下的单次治疗可在治疗后7天内引起微生物群落结构和组成的巨大变化,而多次预约治疗可能会导致口腔菌群多样性的缓慢变化。
    BACKGROUND: The status of dental caries is closely related to changes in the oral microbiome. In this study, we compared the diversity and structure of the dental plaque microbiome in children with severe early childhood caries (S-ECC) before and after general anaesthesia and outpatient treatment.
    METHODS: Forty children aged 3 to 5 years with S-ECC who had completed whole-mouth dental treatment under general anaesthesia (C1) or in outpatient settings (C2) were selected, 20 in each group. The basic information and oral health status of the children were recorded, and the microbial community structure and diversity of dental plaque before treatment (C1, C2), the day after treatment(C2_0D), 7 days after treatment (C1_7D, C2_7D), 1 month after treatment (C1_1M, C2_1M), and 3 months after treatment (C1_3M, C2_3M) were analysed via 16 S rRNA high-throughput sequencing technology.
    RESULTS: (1) The alpha diversity test showed that the flora richness in the multiappointment group was significantly greater at posttreatment than at pretreatment (P < 0.05), and the remaining alpha diversity index did not significantly differ between the 2 groups (P > 0.05). The beta diversity analysis revealed that the flora structures of the C1_7D group and the C2_3M group were significantly different from those of the other time points within the respective groups (P < 0.05). (2) The core flora existed in both the pre- and posttreatment groups, and the proportion of their flora abundance could be altered depending on the caries status of the children in both groups. Leptotrichia abundance was significantly (P < 0.05) lower at 7 days posttreatment in both the single- and multiappointment groups. Corynebacterium and Corynebacterium_matruchotii were significantly more abundant in the C1_1M and C1_3M groups than in the C1 and C1_7D groups (P < 0.05). Streptococcus, Haemophilus and Haemophilus_parainfluenzae were significantly more abundant in the C1_7D group than in the other groups (P < 0.05).
    CONCLUSIONS: A single session of treatment under general anaesthesia can cause dramatic changes in the microbial community structure and composition within 7 days after treatment, whereas treatment over multiple appointments may cause slow changes in oral flora diversity.
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  • 文章类型: Journal Article
    衣原体病,由Chytrid真菌引起的感染性皮肤病,Batrachochytriumdendrobatidis和B.salamandrivorans,对全球两栖动物的生物多样性构成重大威胁。在耐chytrid的两栖动物的皮肤上发现的抗真菌细菌可能会提供针对chytrimycosis的防御和降低耐药个体的死亡率。香港牛顿(Paramesotritonhongkongensis)原产于东亚,一个被怀疑是食糜起源的地区,并表现出无症状感染,表明与食糜的长期共存。因此,这种抗性物种的皮肤微生物群值得调查,以及其他可能影响微生物群的因素。在香港的自然栖息地采样的149只new中,中国,在所有个体中都发现了推定的抗真菌细菌。有314个扩增子序列变体分布在25个推定的抗真菌细菌属;丰富的包括不动杆菌,黄杆菌,和Novoshingoumspp。皮肤微生物群组成受到站点间地理距离的强烈影响。尽管站点间存在差异,我们在不同部位发现了一些核心的皮肤微生物,这些微生物可能对洪孔虫至关重要。占主导地位的核心包括Coamonadaceae家族,科,和Betaproteobacteria类。此外,栖息地升高和宿主性别对皮肤微生物群组成也表现出显着影响。在这些new上发现的抗真菌细菌提供了重要的保护资源,例如开发易感物种的益生菌治疗。
    Chytridiomycosis, an infectious skin disease caused by the chytrid fungi, Batrachochytrium dendrobatidis and B. salamandrivorans, poses a significant threat to amphibian biodiversity worldwide. Antifungal bacteria found on the skin of chytrid-resistant amphibians could potentially provide defense against chytridiomycosis and lower mortality rates among resistant individuals. The Hong Kong newt (Paramesotriton hongkongensis) is native to East Asia, a region suspected to be the origin of chytrids, and has exhibited asymptomatic infection, suggesting a long-term coexistence with the chytrids. Therefore, the skin microbiota of this resistant species warrant investigation, along with other factors that can affect the microbiota. Among the 149 newts sampled in their natural habitats in Hong Kong, China, putative antifungal bacteria were found in all individuals. There were 314 amplicon sequence variants distributed over 25 genera of putative antifungal bacteria; abundant ones included Acinetobacter, Flavobacterium, and Novosphingobium spp. The skin microbiota compositions were strongly influenced by the inter-site geographical distances. Despite inter-site differences, we identified some core skin microbes across sites that could be vital to P. hongkongensis. The dominant cores included the family Comamonadaceae, family Chitinophagaceae, and class Betaproteobacteria. Moreover, habitat elevation and host sex also exhibited significant effects on skin microbiota compositions. The antifungal bacteria found on these newts offer an important resource for conservation against chytridiomycosis, such as developing probiotic treatments for susceptible species.
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  • 文章类型: English Abstract
    由于抗生素的广泛使用,微生物对环境中抗生素的抗性正在增强,抗生素抗性基因(ARGs)的问题越来越严重,严重威胁生态安全和人类健康。为了研究长江口沿海ARGs的分布特征和不同培养基中的微生物群落,通过实地调查收集了八个地点的水和沉积物样本。两个磺胺类抗性基因([STBX]sul1,sul2[STBZ])和六个四环素抗性基因(tetM,tetC,tetX,tetA,tetO,和tetQ),一个整合酶基因intI[STBX]1[STBZ],16SrRNA基因,并对微生物群落进行了检测和分析。结果表明,长江口沿海10个抗性基因的检出率为100%。[STBX]intI1[STBZ]与水样中各种ARGs呈显著正相关。长江口水环境中的细菌门主要是变形杆菌和拟杆菌。叶绿体是水体中的主要细菌,叶绿体和硝基螺旋体是沉积物中的主要细菌属。在水中,Nitrosirota是四个四环素抗性基因(tetX,tetA,tetO,和tetQ)。在沉积物中,Sva0485是[STBX]sul1和intI1[STBZ]共享的潜在宿主社区。微生物群落的分布是影响长江口近岸地区ARGs迁移转化的重要因素。
    Due to the extensive use of antibiotics, the resistance of microorganisms to antibiotics in the environment is increasing, and the problem of antibiotic resistance genes (ARGs) is becoming more and more severe, which seriously threatens ecological security and human health. In order to study the distribution characteristics of ARGs and the microbial community in different media in the coastal area of the Yangtze River Estuary, water and sediment samples from eight sites were collected through a field investigation. Two sulfonamide resistance genes ([STBX]sul1, sul2[STBZ]) and six tetracycline resistance genes (tetM, tetC, tetX, tetA, tetO, and tetQ), one integrase gene intI[STBX]1[STBZ], 16S rRNA gene, and the microbial community were detected and analyzed. The results showed that the detection rate of 10 resistance genes in the coastal area of the Yangtze River Estuary was 100%. [STBX]intI1[STBZ] was significantly positively correlated with various ARGs in the water samples. Proteobacteria and Bacteroidota were the dominant bacteria phyla in the water environment of the Yangtze River Estuary. Chloroplast was the main bacteria genus in water, and Chloroplast and Nitrospira were the main bacteria genera in sediment. In water, Nitrospirota was the common potential host of four tetracycline resistance genes (tetX, tetA, tetO, and tetQ). In sediments, Sva0485 was a potential host community shared by [STBX]sul1 and intI1[STBZ]. The distribution of the microbial community was an important factor affecting the migration and transformation of ARGs in the nearshore area of the Yangtze River Estuary.
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  • 文章类型: Journal Article
    区分腐烂尸体的死前伤害和死后伤害是法医学的难题之一。法医病理学家根据受伤后发生的宏观和微观生命反应来识别死前伤害。然而,分解会使那些重要的反应无效。微生物群因其随时间的推移和对环境变化的敏感性而被广泛用于法医学。在这项研究中,微生物组被引入,以确定细菌群落是否可以用来区分死前和死后的伤害,通过动物实验。我们的发现表明,从伤口产生后的第6天到第9天,细菌群落的差异越来越明显,因为由于分解而无法通过形态学检查确定伤口的类型。属水平的生物标志物可以有效区分损伤类型,其中,仅在死前损伤组中观察到肠球菌和肠杆菌,而葡萄球菌和不动杆菌仅在死后受伤组中。通过检测腐败伤口的细菌群落的差异,可以判断尸体损伤是在死前还是死后发生的。这项研究为死前和死后损伤之间的差异提供了新的视角,并为我们识别死前和死后伤口提供了一种有希望的方法,尤其是腐烂的尸体.
    Differentiating antemortem injury from postmortem injury of decayed cadavers is one of the difficult issues in forensic science. Forensic pathologists identify antemortem injury according to the macroscopic and microscopic vital reactions taken place after being injured. However, the decomposition would render those vital reactions ineffective. Microbiomes have been widely used in forensic science due to their succession with time and sensitivity to vary of environment. In this study, microbiomes were introduced to determine whether the bacterial communities can be used to distinguish between the ante- and postmortem injuries through an animal experiment. Our findings showed that the differences of bacterial community were increasingly apparent from the 6th to 9th day after the wound created when the types of wounds were unidentified by morphological examination due to decomposition. The biomarkers at the genus level could effectively distinguish between injury types, Among them, Enterococcus and Enterobacter were only observed in the antemortem injured group, while Staphylococcus and Acinetobacter were only in the postmortem injured group. It is possible to tell whether cadaveric injuries developed before or after death by detecting differences in the bacterial communities of putrefying wounds. This study provides a new perspective for the differences between ante- and postmortem injuries and provides a promising method for us to identify the ante- and postmortem wounds, especially in decomposed cadavers.
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  • 文章类型: Journal Article
    光自养土壤蓝细菌发挥着重要的生态作用,并且在整个演替早期表现出多样性和丰度的巨大变化。然而,人们对土壤蓝藻群落如何以及为什么随着土壤的发展而变化的了解要少得多,以及植被对这些群落的影响。我们结合了广泛的实地调查,包括跨不同生态系统(从沙漠到热带森林)的16个全球土壤时间序列,通过分子分析来研究光合和非光合土壤蓝细菌的多样性和丰度如何受到土壤发育过程中植被变化的影响,从数百年到数千年的时间段。我们证明,在大多数时间序列中,丰富,随着土壤的发展(从几个世纪到几千年),土壤蓝藻的物种丰富度和群落组成相对稳定。无论土壤年龄如何,森林时间序列始终以非光合蓝细菌(Vampirovibrionia)为主,而草原和灌木以光合蓝藻为主。经历剧烈植被变化(例如从草原到森林的过渡)的时间序列经历了土壤蓝细菌群落组成的显着变化。我们的结果提高了我们对蓝藻类生态学的理解,特别是研究不足的非光合蓝细菌,并强调了植被在土壤发育过程中作为其时间动态的主要驱动力的关键作用。
    Photoautotrophic soil cyanobacteria play essential ecological roles and are known to exhibit large changes in their diversity and abundance throughout early succession. However, much less is known about how and why soil cyanobacterial communities change as soil develops over centuries and millennia, and the effects that vegetation have on such communities. We combined an extensive field survey, including 16 global soil chronosequences across contrasting ecosystems (from deserts to tropical forests), with molecular analyses to investigate how the diversity and abundance of photosynthetic and nonphotosynthetic soil cyanobacteria are affected by vegetation change during soil development, over time periods from hundreds to thousands of years. We show that, in most chronosequences, the abundance, species richness and community composition of soil cyanobacteria are relatively stable as soil develops (from centuries to millennia). Regardless of soil age, forest chronosequences were consistently dominated by nonphotosynthetic cyanobacteria (Vampirovibrionia), while grasslands and shrublands were dominated by photosynthetic cyanobacteria. Chronosequences undergoing drastic vegetation shifts (e.g. transitions from grasslands to forests) experienced significant changes in the composition of soil cyanobacterial communities. Our results advance our understanding of the ecology of cyanobacterial classes, and of the understudied nonphotosynthetic cyanobacteria in particular, and highlight the key role of vegetation as a major driver of their temporal dynamics as soil develops.
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  • 文章类型: Journal Article
    Aims: Permanent treatment of morbid obesity with medication or diet is nearly impossible. Laparoscopic sleeve gastrectomy (LSG) is becoming a widely accepted treatment option. This study profiled and compared gut microbiota composition before and after LSG. Methods & results: A total of 54 stool samples were collected from 27 morbidly obese individuals before and after LSG. The gut microbiota was profiled with 16S amplicon sequencing. After LSG, patients demonstrated a significant decrease (p < 0.001) in BMI and an increase in bacterial diversity. An increased Firmicutes/Bacteroidetes ratio was also noticed after LSG. The families Prevotellaceae and Veillonellaceae predominated in preoperative samples but were markedly lowered after LSG. A marked increase in Akkermansia, Alistipes, Streptococcus, Ruminococcus and Parabacteroides was observed after LSG. Conclusion: In addition to lowering BMI, LSG remodeled gut microbiota composition.
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  • 文章类型: Journal Article
    Anthropogenic activities have inevitably impacted riverine ecosystems, yet their overall contribution to the assemblage of bacterial communities at a large river basin scale remains unclear. In this study, 16S amplicon sequencing was implemented to investigate the bacterial ecosystems in paired water and sediment of North River and West River basins in South China., which contains various anthropogenic environments (e.g., rural/urban area, mining area and livestock area). Subsequently, the links between bacterial community and various types of emerging pollutants in river water were analyzed. The results show that the bacterial assemblage of water and sediment had their own properties that the bacterial community of sediment were mainly affected by seasonal properties, while the bacterial community of water were affected by both seasons and anthropogenic activities. Therein, the aquatic bacterial compositions and abundances were driven by changes in temperature, dissolved oxygen and the emerging pollutants. The dominant phyla Proteobacteria and Firmicutes exhibited adaptability to the mining-affected regions, therein many clades (e.g., Beijerinckiaceae, Acetobacteraceae and Mycobacteriaceae) were also prevalent in the livestock-affected and densely-populated regions. In addition, these two phyla presented associations to the antibiotic resistance in water. The levels of antibiotics, relative antibiotic resistance gens (ARGs) and non-antibiotic pharmaceuticals (NAPs) were closely related to bacterial community composition, diversity and functional diversity, indicating their drive in shifting bacterial communities. Collectively, this work provides a basis for understanding the contribution of anthropogenic activities in shifting bacterial community at a large river basin scale. Further, the results provide new insights for expansion of ecological assessment.
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  • 文章类型: Journal Article
    Wetland ecosystems are critical to the regulation of the global carbon cycle, and there is a high demand for data to improve carbon sequestration and emission models and predictions. Decomposition of plant litter is an important component of ecosystem carbon cycling, yet a lack of knowledge on decay rates in wetlands is an impediment to predicting carbon preservation. Here, we aim to fill this knowledge gap by quantifying the decomposition of standardised green and rooibos tea litter over one year within freshwater and coastal wetland soils across four climates in Australia. We also captured changes in the prokaryotic members of the tea-associated microbiome during this process. Ecosystem type drove differences in tea decay rates and prokaryotic microbiome community composition. Decomposition rates were up to 2-fold higher in mangrove and seagrass soils compared to freshwater wetlands and tidal marshes, in part due to greater leaching-related mass loss. For tidal marshes and freshwater wetlands, the warmer climates had 7-16% less mass remaining compared to temperate climates after a year of decomposition. The prokaryotic microbiome community composition was significantly different between substrate types and sampling times within and across ecosystem types. Microbial indicator analyses suggested putative metabolic pathways common across ecosystems were used to breakdown the tea litter, including increased presence of putative methylotrophs and sulphur oxidisers linked to the introduction of oxygen by root in-growth over the incubation period. Structural equation modelling analyses further highlighted the importance of incubation time on tea decomposition and prokaryotic microbiome community succession, particularly for rooibos tea that experienced a greater proportion of mass loss between three and twelve months compared to green tea. These results provide insights into ecosystem-level attributes that affect both the abiotic and biotic controls of belowground wetland carbon turnover at a continental scale, while also highlighting new decay dynamics for tea litter decomposing under longer incubations.
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  • 文章类型: Journal Article
    微生物在地下生态系统中起着关键作用,喀斯特石漠化(KRD)控制会影响土壤特性和植被覆盖度。然而,KRD控制与土壤细菌群落之间的关系尚不清楚。16SrRNA基因下一代测序用于研究土壤细菌群落结构,composition,多样性,KRD控制区五种生态类型的共现网络。此外,土壤理化性质和土壤化学计量特征,对氮和磷进行了分析。土壤N和P共限制在促进KRD控制对土壤性质的贡献中降低。尽管土壤细菌群落似乎与土壤pH密切相关,土壤钙,土壤磷和植物丰富度,决定它们组成的关键因素是通过改变土壤性质来决定后者。共生网络分析表明,自然生态系统中土壤细菌网络的复杂性高于附加管理生态系统。念珠菌,鸡只杆菌,和Pedosphaeraceae被认为是维持KRD控制区喀斯特土壤生态系统的关键类群。我们的结果表明,自然恢复是退化生态系统恢复和恢复的合适途径。从而有助于不断努力评估土壤-植物生态网络之间的相互作用。
    Microorganisms play critical roles in belowground ecosystems, and karst rocky desertification (KRD) control affects edaphic properties and vegetation coverage. However, the relationship between KRD control and soil bacterial communities remains unclear. 16S rRNA gene next-generation sequencing was used to investigate soil bacterial community structure, composition, diversity, and co-occurrence network from five ecological types in KRD control area. Moreover, soil physical-chemical properties and soil stoichiometry characteristics of carbon, nitrogen and phosphorus were analyzed. Soil N and P co-limitation decreased in the contribution of the promotion of KRD control on edaphic properties. Though soil bacterial communities appeared strongly associated with soil pH, soil calcium, soil phosphorus and plant richness, the key factor to determine their compositions was the latter via changed edaphic properties. The co-occurrence network analysis indicated that soil bacterial network complexity in natural ecosystem was higher than that in additional management ecosystem. Candidatus Udaeobacter, Chthoniobacterales, and Pedosphaeraceae were recognized as the key taxa maintaining karst soil ecosystems in KRD control area. Our results indicate that natural recovery is the suitable way for restoration and rehabilitation of degraded ecosystems, and thus contribute to the ongoing endeavor to appraise the interactions among soil-plant ecological networks.
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