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
    数字皮炎(DD)是一种昂贵的蹄部感染,导致饲养场牛跛行和疼痛。DD病变可以通过一系列临床阶段非线性发展,可以通过Dopfer的M级评分系统进行分类。这种广泛采用的病变评分系统可识别五个DD阶段,其中M1(早期病变),M2(急性溃疡性病变),和M4.1(慢性增生性病变和新发展的病变)被认为是疾病的活跃但独立的阶段。这项研究评估了饲养场牛活性DD病变的皮肤表面微生物群。对来自三个商业饲养场的DD病变进行擦拭,然后根据Dopfer的M期评分系统进行评分。从12个M2和15个M4.1期病变收集拭子样品。来自健康对侧足部(DD对照)的总共21个对照拭子样品被分类为M0期。从没有损伤的完全健康(CH对照)牛收集另外六个皮肤拭子(M0)。使用16S扩增子测序对活性DD病变(M2和M4.1)和健康皮肤(M0)的细菌群落进行剖析。多样性分析表明,M2和M4.1病变的蹄细菌群落均与M0皮肤不同。然而,两个活动病变阶段之间的细菌群落没有差异。与健康皮肤(M0)相比,Spirochaetota和Fusobacteriota的相对丰度显着增加以及细菌多样性的总体减少导致M2和M4.1病变中细菌群落的改变。虽然M2和M4.1阶段被认为是临床上不同的阶段,两个活动阶段的病变相关细菌群落相似.
    Digital dermatitis (DD) is a costly hoof infection, causing lameness and pain in feedlot cattle. DD lesions can develop nonlinearly through a series of clinical stages, which can be classified by Dopfer\'s M-stage scoring system. This widely adopted lesion scoring system recognizes five DD stages, where M1 (early lesion), M2 (acute ulcerative lesion), and M4.1 (chronic proliferative lesion with new developing lesion) are considered active but separate stages of the disease. This study assessed the skin surface microbiota of the active DD lesions of feedlot cattle. The DD lesions from three commercial feedlots were swabbed and then scored according to Dopfer\'s M-stage scoring system. Swab samples were collected from 12 M2- and 15 M4.1-stage lesions. A total of 21 control swab samples from healthy contralateral feet (DD control) were classified as stage M0. An additional six skin swabs (M0) were collected from completely healthy (CH control) cattle with no lesions. The bacterial communities of active DD lesions (M2 and M4.1) and healthy skin (M0) were profiled using 16S amplicon sequencing. Diversity analyses showed that the hoof bacterial communities of M2 and M4.1 lesions were each distinct from those of M0 skin. However, the bacterial communities between the two active lesion stages were not different from each other. A significant increase in the relative abundance of Spirochaetota and Fusobacteriota and an overall decrease in bacterial diversity contributed to the altered bacterial communities in M2 and M4.1 lesions compared to those of healthy skin (M0). Although stages M2 and M4.1 are considered clinically different stages, the lesion-associated bacterial community is similar between the two active stages.
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  • 文章类型: Journal Article
    浸出床反应器(LBR)是干式厌氧系统,可以处理高固体含量的原料,比如鸡粪,添加最少的水。在这项研究中,鸡粪与沸石混合,一个新颖的补充,并包装在LBR中,以提高沼气产量。然后在连续搅拌釜反应器(CSTR)中处理所得的渗滤液,大部分甲烷是在那里产生的。将CSTR的上清液返回到LBR。LBR的分批模式操作导致甲烷生产率(MPR)变化,当渗滤液富含有机物时,在每个分批循环开始时达到峰值。比较两个系统中的MPR,沸石系统中的峰比控制系统中的峰更高且更尖锐,在压力下,如2328mgL-1的乙酸积累所示。此外,沸石在LBR中的存在起着至关重要的作用,在固体停留时间为14d时,将每克鸡粪挥发性固体的总甲烷产量从0.142(对照实验)提高到0.171NLCH4。沸石还提高了系统的稳定性。由于进入系统的水很少,氨浓度逐渐增加,在实验结束时达到3220mgL-1(控制系统)和2730mgL-1(沸石系统)。与对照实验(17.3mgL-1d-1)相比,沸石似乎以较低的速率(14.0mgL-1d-1)促进氨的积累。对来自用沸石修正的LBR的渗滤液的CSTR的微生物分析显示,与对照实验(69.1%)相比,甲烷的相对丰度更高(83.6%)。两种CSTR在手术120天后建立了与接种物显著不同的细菌谱(p<0.05)。关于考古社区,CSTRs与接种物之间无显著统计学差异(p>0.05)。
    Leach bed reactors (LBRs) are dry anaerobic systems that can handle feedstocks with high solid content, like chicken manure, with minimal water addition. In this study, the chicken manure was mixed with zeolite, a novel addition, and packed in the LBR to improve biogas production. The resulting leachate was then processed in a continuous stirred tank reactor (CSTR), where most of the methane was produced. The supernatant of the CSTR was returned to the LBR. The batch mode operation of the LBR led to a varying methane production rate (MPR) with a peak in the beginning of each batch cycle when the leachate was rich in organic matter. Comparing the MPR in both systems, the peaks in the zeolite system were higher and more acute than in the control system, which was under stress, as indicated by the acetate accumulation at 2328 mg L-1. Moreover, the presence of zeolite in the LBR played a crucial role, increasing the overall methane yield from 0.142 (control experiment) to 0.171 NL CH4 per g of volatile solids of chicken manure entering the system at a solid retention time of 14 d. Zeolite also improved the stability of the system. The ammonia concentration increased gradually due to the little water entering the system and reached 3220 mg L-1 (control system) and 2730 mg L-1 (zeolite system) at the end of the experiment. It seems that zeolite favored the accumulation of the ammonia at a lower rate (14.0 mg L-1 d-1) compared to the control experiment (17.3 mg L-1 d-1). The microbial analysis of the CSTR fed on the leachate from the LBR amended with zeolite showed a higher relative abundance of Methanosaeta (83.6%) compared to the control experiment (69.1%). Both CSTRs established significantly different bacterial profiles from the inoculum after 120 days of operation (p < 0.05). Regarding the archaeal communities, there were no significant statistical differences between the CSTRs and the inoculum (p > 0.05).
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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
    以前,我们表明,在哈沙湖水样中预防性添加葡萄糖可以抑制蓝藻的生长,至少在短时间内。当前的研究测试了整个哈沙湖开花季节的葡萄糖控制蓝藻。从6月9日开始并持续到2022年8月24日的藻华季节,每周从Harsha湖收集水样(1000ml)。两个7升聚丙烯容器中的每一个,加入500毫升的哈沙湖水,并将容器放置在受控环境室中。到一个标有“已处理”的容器,“添加0.15克葡萄糖,并且没有添加到标记为“控件”的容器中。\"之后,从每个容器中收集3份25ml样本,每周用于16SrRNA基因测序.然后每周新收集1000毫升的哈沙湖水,每个容器中加入500毫升,同时向“处理过的”容器中添加0.15g葡萄糖。测序数据用于检查处理和对照容器之间细菌群落组成的差异。用葡萄糖处理通过以下方式改变了微生物群落:1)减少分类多样性,2)在很大程度上消除了蓝藻类群,和3)增加非蓝细菌类群(如变形杆菌和放线菌群)的亚群的相对丰度。尽管每周都有直接来自湖水的输入,但这些影响还是在不同时间观察到的。在整个夏季盛开季节,向每周增加湖水的容器中添加葡萄糖会抑制蓝细菌种群。葡萄糖似乎以蓝细菌为代价刺激了某些细菌类群的多样性。
    Previously, we showed that prophylactic addition of glucose to Harsha Lake water samples could inhibit cyanobacteria growth, at least for a short period of time. The current study tested cyanobacterial control with glucose for the entire Harsha Lake bloom season. Water samples (1000 ml) were collected weekly from Harsha Lake during the algal-bloom season starting June 9 and lasting until August 24, 2022. To each of two 7-liter polypropylene containers, 500 ml of Harsha Lake water was added, and the containers were placed in a controlled environment chamber. To one container labeled \"Treated,\" 0.15 g of glucose was added, and nothing was added to the container labeled \"Control.\" After that, three 25 ml samples from each container were collected and used for 16S rRNA gene sequencing each week. Then 1000 ml of Harsha Lake water was newly collected each week, with 500 ml added to each container, along with the addition of 0.15 g glucose to the \"Treated\" container. Sequencing data were used to examine differences in the composition of bacterial communities between Treated and Control containers. Treatment with glucose altered the microbial communities by 1) reducing taxonomic diversity, 2) largely eliminating cyanobacterial taxa, and 3) increasing the relative abundance of subsets of non-cyanobacterial taxa (such as Proteobacteria and Actinobacteriota). These effects were observed across time despite weekly inputs derived directly from Lake water. The addition of glucose to a container receiving weekly additions of Lake water suppressed the cyanobacterial populations during the entire summer bloom season. The glucose appears to stimulate the diversity of certain bacterial taxa at the expense of the cyanobacteria.
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  • 文章类型: Journal Article
    2011年3月东北太平洋地震引发海啸后,福岛第一核电站发生了一起重大事件,海水进入反应堆大楼地下室的环面室。这里,我们鉴定和分析了圆环室内水和几个环境样品中的细菌群落。东京电力公司控股公司从距离房间底部(TW1)和底层(TW2)30cm至1m之间的两个采样点收集了环形房间水(1×109Bq137Cs/L)。基于16SrRNA扩增子测序的细菌群落结构分析表明,TW1和TW2中的主要细菌属相似。TW1主要包含Limnobacter属,硫代硫酸盐氧化细菌。氧化硫脂林杆菌的γ辐照试验,在TW1中发现的最密切相关的系统发育学表明,其抗辐射能力与普通细菌相似。TW2主要包含Brevirhabdus属,锰氧化细菌.尽管圆环室水中的细菌多样性低于福岛附近的海水,约70%的鉴定属与金属腐蚀有关。潜在环境分配-一种估计栖息地分布和共同检测分析的分析技术-揭示了环面室内水中的微生物群落起源于天然海洋微生物和典型生物膜的人工细菌群落的独特混合物,污泥,和废水。了解与受损植物中金属腐蚀有关的特定细菌对于推进退役工作非常重要。
    目的:在核电站退役的背景下,微生物在反应器和管道系统内的增殖构成了巨大的挑战。因此,在这样的环境中,微生物群落的识别是至关重要的。在福岛第一核电站事故之后,微生物群落分析是对主要在场地外收集的环境样品进行的。然而,使用现场区域的样本进行分析,包括邻近的土壤和海水,没有执行。这项研究代表了对微生物群落的首次全面分析,利用meta16S扩增子测序,重点关注从圆环室内含放射性元素的水中收集的环境样本,包括周围环境。一些已确定的微生物属与以前在法国和巴西等国家的乏核燃料池中确定的微生物属共享。此外,我们在本文中的讨论阐明了许多这些细菌与金属腐蚀的相关性。
    A major incident occurred at the Fukushima Daiichi Nuclear Power Station following the tsunami triggered by the Tohoku-Pacific Ocean Earthquake in March 2011, whereby seawater entered the torus room in the basement of the reactor building. Here, we identify and analyze the bacterial communities in the torus room water and several environmental samples. Samples of the torus room water (1 × 109 Bq137Cs/L) were collected by the Tokyo Electric Power Company Holdings from two sampling points between 30 cm and 1 m from the bottom of the room (TW1) and the bottom layer (TW2). A structural analysis of the bacterial communities based on 16S rRNA amplicon sequencing revealed that the predominant bacterial genera in TW1 and TW2 were similar. TW1 primarily contained the genus Limnobacter, a thiosulfate-oxidizing bacterium. γ-Irradiation tests on Limnobacter thiooxidans, the most closely related phylogenetically found in TW1, indicated that its radiation resistance was similar to ordinary bacteria. TW2 predominantly contained the genus Brevirhabdus, a manganese-oxidizing bacterium. Although bacterial diversity in the torus room water was lower than seawater near Fukushima, ~70% of identified genera were associated with metal corrosion. Latent environment allocation-an analytical technique that estimates habitat distributions and co-detection analyses-revealed that the microbial communities in the torus room water originated from a distinct blend of natural marine microbial and artificial bacterial communities typical of biofilms, sludge, and wastewater. Understanding the specific bacteria linked to metal corrosion in damaged plants is important for advancing decommissioning efforts.
    OBJECTIVE: In the context of nuclear power station decommissioning, the proliferation of microorganisms within the reactor and piping systems constitutes a formidable challenge. Therefore, the identification of microbial communities in such environments is of paramount importance. In the aftermath of the Fukushima Daiichi Nuclear Power Station accident, microbial community analysis was conducted on environmental samples collected mainly outside the site. However, analyses using samples from on-site areas, including adjacent soil and seawater, were not performed. This study represents the first comprehensive analysis of microbial communities, utilizing meta 16S amplicon sequencing, with a focus on environmental samples collected from the radioactive element-containing water in the torus room, including the surrounding environments. Some of the identified microbial genera are shared with those previously identified in spent nuclear fuel pools in countries such as France and Brazil. Moreover, our discussion in this paper elucidates the correlation of many of these bacteria with metal corrosion.
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  • 文章类型: Journal Article
    背景:跛行被定义为由于运动系统功能障碍而改变或异常的步态,是饲养场牛的健康问题,有重大的经济,劳动,和福利影响。数字皮炎(足足底表面的DD-病变)和脚腐烂(FR-影响指间裂隙)是饲养场跛行的常见感染原因。这些蹄病变可以单独或组合(DDFR)在同一蹄中发生。从位于艾伯塔省南部的三个商业饲养场总共收集了208个蹄拭子。每个损伤样本与取自健康对侧足部的相应对照皮肤样本相匹配。还从任何脚上没有损伤的牛收集对照皮肤样品。三种蹄病变的细菌群落(DD,DD+FR,FR)和健康皮肤使用16S扩增子测序进行轮廓分析。
    结果:Alpha多样性分析显示,与对照皮肤相比,DD和FR病变上的细菌多样性较低。β多样性分析表明,DD的细菌群落,FR,DD+FR病变与对照皮肤不同。虽然饲养场的影响很小,病变类型占细菌群落变化的22%(PERMANOVA-R=0.22,P<0.01)。与相应的对照皮肤相比,DD中有11、12和3个差异丰富(DA)细菌属,DD+FR,和FR病变,分别。
    结论:DD+FR病变的细菌群落描述是一个新发现。与健康皮肤相比,病变不仅导致细菌群落改变,但是这些群落的组成也因蹄部病变而异。表面拭子的16S扩增子测序作为分离不同蹄病变的研究工具具有重要价值,并且可以为饲养场牛DD和FR的多细菌病因提供更多见解。
    BACKGROUND: Lameness is defined as altered or abnormal gait due to dysfunction of the locomotor system, and is a health issue of feedlot cattle, having major economic, labour, and welfare implications. Digital dermatitis (DD-a lesion of the plantar surface of the foot) and foot rot (FR-affects the interdigital cleft) are common infectious causes of lameness in feedlots. These hoof lesions can occur alone or in combination (DD + FR) in the same hoof. A total of 208 hoof swabs were collected from three commercial feedlots located in southern Alberta. Every lesion sample was matched with a corresponding control skin sample taken from a healthy contralateral foot. Control skin samples were also collected from cattle with no lesion on any feet. Bacterial communities of three types of hoof lesions (DD, DD + FR, FR) and healthy skin were profiled using 16S amplicon sequencing.
    RESULTS: Alpha diversity analysis revealed a lower bacterial diversity on DD and FR lesions compared to control skin. Beta diversity analysis showed that bacterial communities of DD, FR, and DD + FR lesions were distinct from those of the control skin. While the impact of feedlot was minimal, lesion type contributed to 22% of the variation observed among bacterial communities (PERMANOVA-R = 0.22, P < 0.01). Compared to the corresponding control skin, there were 11, 12, and 3 differentially abundant (DA) bacterial genera in DD, DD + FR, and FR lesions, respectively.
    CONCLUSIONS: The bacterial community description of a DD + FR lesion is a novel finding. Not only did lesions lead to altered bacterial communities when compared to healthy skin, but the composition of those communities also differed depending on the hoof lesion. The 16S amplicon sequencing of surface swabs has significant value as a research tool in separating different hoof lesions and can provide additional insights to the polybacterial etiology of DD and FR in feedlot cattle.
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  • 文章类型: Journal Article
    尽管微生物降解是表层海水中多环芳烃(PAH)的关键汇,缺乏基于野外的证据,表明生物降解的区域差异以及PAHs对特定地点微生物群落的影响。我们比较了PAH降解的程度及其在地中海沿岸和南极海洋微生物群的短期孵化中的影响与环境相关的PAHs浓度。地中海细菌容易降解疏水性较低的PAHs,速率平均为4.72±0.5ngLh-1。代谢组学反应显示与水平基因转移相关的基因显着富集,应激反应,和PAH降解,主要由阿尔法变形杆菌包藏。群落组成发生变化,并增加了拟杆菌和黄杆菌的相对丰度。在南极水域,没有PAH的降解,观察到最小的转移组反应。这些结果为地理区域等因素提供了证据,社区组成,和暴露前历史来预测海水中PAH的生物降解。
    Although microbial degradation is a key sink of polycyclic aromatic hydrocarbons (PAH) in surface seawaters, there is a dearth of field-based evidences of regional divergences in biodegradation and the effects of PAHs on site-specific microbial communities. We compared the magnitude of PAH degradation and its impacts in short-term incubations of coastal Mediterranean and the Maritime Antarctica microbiomes with environmentally relevant concentrations of PAHs. Mediterranean bacteria readily degraded the less hydrophobic PAHs, with rates averaging 4.72 ± 0.5 ng L h-1. Metatranscriptomic responses showed significant enrichments of genes associated to horizontal gene transfer, stress response, and PAH degradation, mainly harbored by Alphaproteobacteria. Community composition changed and increased relative abundances of Bacteroidota and Flavobacteriales. In Antarctic waters, there was no degradation of PAH, and minimal metatranscriptome responses were observed. These results provide evidence for factors such as geographic region, community composition, and pre-exposure history to predict PAH biodegradation in seawater.
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  • 文章类型: Journal Article
    背景:使用常规培养方法正确鉴定威胁肢体的糖尿病足溃疡(LTDFU)患者的多微生物微生物是不够的。与单独的标准治疗相比,这项前瞻性研究评估了辅助分子测试在LTDFU中识别挑剔微生物的潜在价值。
    方法:90例LTDFU患者在转诊糖尿病足中心接受了跨学科和标准的抗生素治疗。同时使用16S扩增子测序(16SAS)标本以及入院时收集的常规培养物来回顾性评估微生物学发现及其与截肢结局的关联。
    结果:16SAS显示的微生物计数超过了常规培养(17vs.3个细菌/溃疡分别)。窄食单胞菌属。在29例患者中发现与严重(踝关节以上)截肢高度相关(OR:4.76,95%CI1.01-22.56),而只有一种是通过常规培养同时鉴定的。因此,有27例患者在治疗期间没有适当的抗生素覆盖。
    结论:辅助分子检测有助于鉴定严重的病原体,例如寡养单胞菌感染,并可能与LTDFU患者的严重截肢有关。
    BACKGROUND: Proper identification of the polymicrobial microorganisms in patients with limb-threatening diabetic foot ulcers (LTDFUs) using conventional culture is insufficient. This prospective study evaluates the potential value of adjuvant molecular testing assisting in identify fastidious micro-organisms in LTDFUs compared to standard treatment alone.
    METHODS: Ninety patients with LTDFUs received interdisciplinary and standard antibiotic treatment in a referral diabetic foot center. A simultaneous 16S amplicon sequencing (16S AS) specimen along with conventional culture collected at admission was used to retrospectively evaluate the microbiological findings and its association with amputation outcomes.
    RESULTS: The microorganism count revealed by 16S AS overwhelmed that of conventional culturing (17 vs. 3 bacteria/ulcer respectively). The Stenotrophomonas spp. revealed in 29 patients were highly correlated with major (above ankle) amputation (OR: 4.76, 95% CI 1.01-22.56), while only one had been concomitantly identified by conventional culturing. Thus, there were 27 cases without proper antibiotics coverage during treatment.
    CONCLUSIONS: Adjuvant molecular testing assisted identification of fastidious pathogens such as Stenotrophomonas infection and might be associated with major amputation in patients with LTDFUs.
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  • 文章类型: Preprint
    了解塑造人类微生物组变化的因素是生物学研究的主要目标。虽然其他基因组学领域已经使用了大量,预编译的简编,以提取系统的见解,否则需要不切实际的样本量,对于通常用于量化微生物组组成的16SrRNA测序数据,还没有可比的资源。为了帮助缩小这个差距,我们已经收集了一组168,484个公开的人类肠道微生物组样本,用单一管道处理,并组合成迄今为止最大的统一微生物组数据集。我们利用这个资源,可以在microbiomap.org上免费获得,揭示人类肠道微生物组的全球变化。我们发现Firmicutes,尤其是芽孢杆菌和梭菌,几乎普遍存在于人类肠道中。同时,65个最常见的微生物属的相对丰度在至少两个世界地区之间存在差异。我们还表明,样本不足的世界地区的肠道微生物组,例如中亚和南亚,与欧洲和北美特征更彻底的微生物区有很大不同。此外,这些被忽视地区的人类可能藏有数百个由于这种采样不足而尚未发现的分类群,强调微生物组研究中多样性的必要性。我们预计这一新的纲要可以为社区服务,并实现先进的应用和方法学研究。
    Understanding the factors that shape variation in the human microbiome is a major goal of research in biology. While other genomics fields have used large, pre-compiled compendia to extract systematic insights requiring otherwise impractical sample sizes, there has been no comparable resource for the 16S rRNA sequencing data commonly used to quantify microbiome composition. To help close this gap, we have assembled a set of 168,484 publicly available human gut microbiome samples, processed with a single pipeline and combined into the largest unified microbiome dataset to date. We use this resource, which is freely available at microbiomap.org, to shed light on global variation in the human gut microbiome. We find that Firmicutes, particularly Bacilli and Clostridia, are almost universally present in the human gut. At the same time, the relative abundance of the 65 most common microbial genera differ between at least two world regions. We also show that gut microbiomes in undersampled world regions, such as Central and Southern Asia, differ significantly from the more thoroughly characterized microbiomes of Europe and Northern America. Moreover, humans in these overlooked regions likely harbor hundreds of taxa that have not yet been discovered due to this undersampling, highlighting the need for diversity in microbiome studies. We anticipate that this new compendium can serve the community and enable advanced applied and methodological research.
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