16s rRNA sequencing

16S rRNA 测序
  • 文章类型: Journal Article
    本研究旨在探讨补充胆汁酸对脂肪肝出血性综合征(FLHS)的影响,生产性能,蛋鸡的生理和品质特性。60只三黄蛋鸡,年龄28周,在4周内被随机分配到6种饮食治疗,包括对照(CON)组(饲喂基础饮食),高脂饮食(HFD)治疗组(含10%大豆油的基础饮食),和补充有0.01%和0.02%的鹅去氧胆酸(CDCA)或猪去氧胆酸(HDCA)组的HFD。生产性能,鸡蛋质量,肝脏形态学,血清生化指标,抗氧化能力,促炎细胞因子,和肠道微生物群进行了评估。0.01%CDCA中的平均体重大于HFD组(p<0.05)。CON组的蛋壳厚度大于HFD,0.01%CDCA,和HDCA组(p<0.05)。0.02%HDCA组的卵白高度高于HFD组(p<0.05)。HFD组的蛋壳重量小于CON组(p<0.05)。HDCA组Haugh单位(HU)大于HFD组(p<0.05)。0.02%HDCA组的白蛋白重量大于CON和HFD组(p<0.05)。在HFD组中,甘油三酯(TG)的水平,总胆固醇(TC),低密度脂蛋白胆固醇(LDL-C)高于其他组(p<0.05)。HFD组过氧化氢酶(CAT)和总超氧化物歧化酶(T-SOD)水平低于其他组(p<0.05)。HFD组丙二醛(MDA)水平高于其他组(p<0.05)。HFD组的肿瘤坏死因子-α(TNF-α)水平高于其他组(p<0.05)。16SrRNA测序分析表明不同处理组之间特定细菌种群的相对丰度存在显著差异。治疗组和CON组表现出抑制宿主能量吸收或促进肠道健康的细菌,如Firmicutes,拟杆菌,和Ruminococus,而HFD组显示潜在致病性或有害细菌的患病率增加,如Desulfovibriospp.总之,已证明在家禽饲料中补充BA可有效减轻FLHS对蛋鸡的有害影响。这种干预调节脂质代谢,加强抗氧化防御,减少炎症,调节肠道微生物群,为BAs在家禽业中的应用提供了一个新的视角。
    This study aimed to investigate the effects of bile acids (BAs) supplementation on fatty liver hemorrhagic syndrome (FLHS), production performance, and physiological and quality characteristics of laying hen eggs. Sixty Sanhuang laying hens, aged 28 weeks, were randomly allocated to six dietary treatments over a 4-week period, including the control (CON) group (feeding basal diet), the high-fat diet (HFD)-treated group (basal diet containing 10% soybean oil), and HFD supplemented with 0.01% and 0.02% of chenodeoxycholic acid (CDCA) or hyodeoxycholic acid (HDCA) groups. Production performance, egg quality, liver morphology, serum biochemical indexes, antioxidant capacity, proinflammatory cytokines, and intestinal microbiota were evaluated. The average body weight in 0.01% CDCA was larger than in the HFD group (p < 0.05). Eggshell Thickness in the CON group was greater than in the HFD, 0.01% CDCA, and HDCA groups (p < 0.05). Albumen height in the 0.02% HDCA group was higher than the HFD group (p < 0.05). Eggshell weight in the HFD group was less than the CON group (p < 0.05). Haugh unit (HU) in the HDCA group was larger than the HFD group (p < 0.05). Albumen weight in the 0.02% HDCA group was greater than the CON and HFD groups (p < 0.05). In the HFD group, the levels of triglyceride (TG), total cholesterol (TC), and low-density lipo-protein cholesterol (LDL-C) were surpassing the other groups (p < 0.05). The levels of catalase (CAT) and total superoxide dismutase (T-SOD) in the HFD group was smaller than the other groups (p < 0.05). The level of malondialdehyde (MDA) in the HFD group was higher than in the other groups (p < 0.05). Tumor necrosis factor-α (TNF-α) levels were larger in the HFD group than in the other groups (p < 0.05). The 16S rRNA sequencing analysis indicated significant variations in the relative abundance of specific bacterial populations among the different treatment groups. The treatment and CON groups exhibited a higher presence of bacteria that inhibit host energy absorption or promote intestinal health such as Firmicutes, Bacteroidetes, and Ruminococcus, whereas the HFD group showed an increased prevalence of potentially pathogenic or deleterious bacteria, such as Desulfovibrio spp. In conclusion, the supplementation of BAs in poultry feed has been demonstrated to effectively mitigate the detrimental effects of FLHS in laying hens. This intervention regulates lipid metabolism, bolsters antioxidant defenses, reduces inflammation, and modulates the gut microbiota, offering a novel perspective on the application of BAs in the poultry industry.
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  • 文章类型: Journal Article
    地热特征,如温泉和泥火山,寄主不同的微生物生命,包括许多极端微生物。地热特征的物理化学参数,如温度,pH值,和重金属浓度,可以影响这些环境中微生物生命的α和β多样性,站点和采样之间的时空差异也是如此。在这项研究中,从黄石国家公园的八个地热地点收集和分析了水和沉积物样本,包括六个温泉,一座泥火山,在2019年7月的同一周内,一个酸性湖泊,这些地热地点的温度变化很大,pH值,和化学成分。用相同的方法和分类学谱以及用16SrRNA测序确定的α和β多样性度量来处理和分析所有样品。然后分析这些微生物多样性结果与pH值,温度,和地热特征的化学成分。结果表明,根据地热场地的物理化学组成,主要的微生物种类变化很大,随着pH值的降低和水中溶解重金属的增加,对应于α多样性的降低,尤其是在沉积物样本中.同样,与相对中性或碱性pH的地热站点相比,具有酸性pH值的站点彼此之间具有更多相似的微生物种群(β多样性)。这项研究表明,与这些地点的温度相比,pH和/或重金属浓度是微生物多样性和种群分布的更重要的驱动因素,也是黄石国家公园多个地热地点首次报道的微生物多样性研究。包括相对较新的泥火山黑龙Caldron,它在1948年爆发。
    Geothermal features, such as hot springs and mud volcanoes, host diverse microbial life, including many extremophile organisms. The physicochemical parameters of the geothermal feature, such as temperature, pH, and heavy metal concentration, can influence the alpha and beta diversity of microbial life in these environments, as can spatiotemporal differences between sites and sampling. In this study, water and sediment samples were collected and analyzed from eight geothermal sites at Yellowstone National Park, including six hot springs, a mud volcano, and an acidic lake within the same week in July 2019, and these geothermal sites varied greatly in their temperature, pH, and chemical composition. All samples were processed and analyzed with the same methodology and taxonomic profiles and alpha and beta diversity metrics determined with 16S rRNA sequencing. These microbial diversity results were then analyzed with respect to pH, temperature, and chemical composition of the geothermal features. Results indicated that predominant microbial species varied greatly depending on the physicochemical composition of the geothermal site, with decreases in pH and increases in dissolved heavy metals in the water corresponding to decreases in alpha diversity, especially in the sediment samples. Similarly, sites with acidic pH values had more similar microbial populations (beta diversity) to one another than to relatively neutral or alkaline pH geothermal sites. This study suggests that pH and/or heavy metal concentration is a more important driver for microbial diversity and population profile than the temperature for these sites and is also the first reported microbial diversity study for multiple geothermal sites in Yellowstone National Park, including the relatively new mud volcano Black Dragon\'s Caldron, which erupted in 1948.
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  • 文章类型: Journal Article
    银屑病是一种常见的慢性炎症性皮肤病。牛皮癣的出现与皮肤表面微生物群的生态失调和免疫微环境的失衡有关。在这项研究中,我们研究了局部胸腺五肽(TP5)对咪喹莫特(IMQ)诱导的小鼠银屑病的治疗影响,以及TP5对皮肤免疫环境和皮肤表面微生物群的调节作用。
    使用IMQ诱导的银屑病样病变小鼠模型来鉴定TP5的靶标和分子机制。免疫荧光用于鉴定TP5治疗前后T细胞亚群表达的差异。使用蛋白质印迹(WB)评估NF-κB信号传导途径组分的表达的变化。16SrRNA测序和网络药理学用于检测TP5给药前后皮肤菌群的变化。
    体内,TP5减少了IMQ诱导的小鼠背部炎症。H&E染色显示表皮厚度减少和TP5的炎性细胞浸润。Masson染色显示TP5给药后表皮和真皮胶原蛋白浸润减少。免疫组织化学显示TP5处理显著降低IL-17表达。免疫浸润分析的结果显示牛皮癣性病变具有更多的T细胞亚群。根据免疫荧光结果,TP5显著降低了CD4+的比例,Th17,ROR+,和CD8+T细胞。WB显示TP5降低了IMQ诱导的银屑病模型小鼠皮肤组织中NF-κB通路的表达。16SrRNA测序显示伯克霍尔德菌和假单胞菌科假单胞菌显着增加,葡萄球菌科葡萄球菌显着减少,水细菌,Herbaspirillum,和Balneimonas。TP5处理后,Firmicutes主导了皮肤微生物多样性,而拟杆菌,Verrucomicrobia,TM7,变形杆菌,放线菌,酸杆菌,双子座,和其他物种在IMQ组中占主导地位。
    TP5可能通过调节表皮菌群来治疗牛皮癣,降低NF-κB通路表达,影响T细胞亚群。
    UNASSIGNED: Psoriasis is a common chronic inflammatory skin condition. The emergence of psoriasis has been linked to dysbiosis of the microbiota on the skin surface and an imbalance in the immunological microenvironment. In this study, we investigated the therapeutic impact of topical thymopentin (TP5) on imiquimod (IMQ)-induced psoriasis in mice, as well as the modulatory influence of TP5 on the skin immune milieu and the skin surface microbiota.
    UNASSIGNED: The IMQ-induced psoriasis-like lesion mouse model was used to identify the targets and molecular mechanisms of TP5. Immunofluorescence was employed to identify differences in T-cell subset expression before and after TP5 therapy. Changes in the expression of NF-κB signaling pathway components were assessed using Western blotting (WB). 16S rRNA sequencing and network pharmacology were used to detect changes in the skin flora before and after TP5 administration.
    UNASSIGNED: In vivo, TP5 reduced IMQ-induced back inflammation in mice. H&E staining revealed decreased epidermal thickness and inflammatory cell infiltration with TP5. Masson staining revealed decreased epidermal and dermal collagen infiltration after TP5 administration. Immunohistochemistry showed that TP5 treatment dramatically reduced IL-17 expression. Results of the immunoinfiltration analyses showed psoriatic lesions with more T-cell subsets. According to the immunofluorescence results, TP5 dramatically declined the proportions of CD4+, Th17, ROR+, and CD8+ T cells. WB revealed that TP5 reduced NF-κB pathway expression in skin tissues from IMQ-induced psoriasis model mice. 16S rRNA sequencing revealed a significant increase in Burkholderia and Pseudomonadaceae_Pseudomonas and a significant decrease in Staphylococcaceae_Staphylococcus, Aquabacterium, Herbaspirillum, and Balneimonas. Firmicutes dominated the skin microbial diversity after TP5 treatment, while Bacteroidetes, Verrucomicrobia, TM7, Proteobacteria, Actinobacteria, Acidobacteria, Gemmatimonadetes, and other species dominated in the IMQ group.
    UNASSIGNED: TP5 may treat psoriasis by modulating the epidermal flora, reducing NF-κB pathway expression, and influencing T-cell subsets.
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  • 文章类型: Journal Article
    本研究旨在探讨视网膜静脉阻塞(RVO),失明的眼部状况,以及肠道微生物群组成的改变,提供对RVO发病机制的见解。使用16SrRNA测序和液相色谱-质谱(LC-MS)分析来自25名RVO患者和11名非RVO个体的粪便样品。在RVO和非RVO组之间注意到肠道微生物物种的丰度的显著差异。在门一级,RVO组显示厚壁菌与拟杆菌的比例升高。在属一级,与非RVO组相比,RVO组表现出更高的大肠杆菌_志贺氏菌丰度(P<0.05),而副杆菌属丰度较低(P<0.01)。功能预测表明叶酸合成减少,生物素代谢,和氧化磷酸化,随着RVO组丁酸代谢的增加。LC-MS分析显示嘌呤代谢存在显著差异,ABC运输商,和萘降解途径,尤其是嘌呤代谢。Pearson相关性分析显示细菌属和粪便代谢产物之间存在显著关联。富集分析强调了特定代谢物和细菌属之间的联系。研究结果表明,在RVO患者中观察到肠道菌群失调,提示肠道微生物群是潜在的治疗靶点。调节肠道微生物群可能是管理RVO和改善患者预后的新策略。此外,研究结果表明,肠道微生物菌群失调参与RVO的发育,强调了解其发病机制对有效治疗发展的重要性。
    目的:视网膜静脉阻塞(RVO)是一种致盲眼病,了解其发病机制对于开发有效的治疗方法至关重要。这项研究表明,RVO患者和非RVO个体之间的肠道菌群组成存在显着差异。肠道微生物菌群失调参与RVO发育。功能预测和代谢分析提供了对潜在机制的见解,突出治疗干预的潜在途径。这些发现表明,调节肠道微生物群可能是管理RVO和改善患者预后的有希望的策略。
    This study aims to explore the link between retinal vein occlusion (RVO), a blinding ocular condition, and alterations in gut microbiota composition, to offer insights into the pathogenesis of RVO. Fecal samples from 25 RVO patients and 11 non-RVO individuals were analyzed using 16S rRNA sequencing and liquid chromatography-mass spectrometry (LC-MS). Significant differences in the abundance of gut microbial species were noted between RVO and non-RVO groups. At the phylum level, the RVO group showed an elevation in the ratio of Firmicutes to Bacteroidetes. At the genus level, the RVO group showed higher abundance in Escherichia_Shigella (P < 0.05) and less abundance in Parabacteroides (P < 0.01) than the non-RVO group. Functional predictions indicated reduced folate synthesis, biotin metabolism, and oxidative phosphorylation, with an increase in butyric acid metabolism in the RVO group. LC-MS analysis showed significant differences in purine metabolism, ABC transporters, and naphthalene degradation pathways, especially purine metabolism. Pearson correlation analysis revealed significant associations between bacterial genera and fecal metabolites. Enrichment analysis highlighted connections between specific metabolites and bacterial genera. The findings showed that the dysregulation of gut microbiota was observed in RVO patients, suggesting the gut microbiota as a potential therapeutic target. Modulating the gut microbiota could be a novel strategy for managing RVO and improving patient outcomes. Furthermore, the study findings suggest the involvement of gut microbial dysbiosis in RVO development, underscoring the significance of understanding its pathogenesis for effective treatment development.
    OBJECTIVE: Retinal vein occlusion (RVO) is a blinding ocular condition, and understanding its pathogenesis is crucial for developing effective treatments. This study demonstrates significant differences in gut microbiota composition between RVO patients and non-RVO individuals, implicating the involvement of gut microbial dysbiosis in RVO development. Functional predictions and metabolic profiling provide insights into the underlying mechanisms, highlighting potential pathways for therapeutic intervention. These findings suggest that modulating the gut microbiota might be a promising strategy for managing RVO and improving patient outcomes.
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  • 文章类型: Journal Article
    最近,越来越多的报道描述了动物来源的病原体,这些病原体引起各种感染,并增加了它们向人类的传播。溶胆链球菌,牛链球菌/马链球菌复合体(SBSEC)的成员,是这些病原体之一,感染从哺乳动物到家禽的多种宿主,并且具有从致病性到食物发酵的广泛功能。由于胆溶菌引起包括菌血症在内的并发症,感染性心内膜炎,和人类的结肠直肠恶性肿瘤,重要的是调查它在各种宿主中的发生,包括鹅,以防止潜在的人畜共患传播。这项研究旨在调查临床健康和腹泻的鹅粪便中是否存在胆溶菌,密集和半密集地饲养,通过体外培养方法,通过PCR和基于序列的分子方法表征回收的分离株,并通过圆盘扩散和梯度测试方法确定其抗生素敏感性。为此,使用150个新鲜鹅粪便样品。对卵溶链球菌的培养阳性被确定为8%(12/150)。PCR分析鉴定出54.55%(n=6)的分离株为溶胆囊链球菌亚种。胆溶菌和45.45%(n=5)为胆溶菌亚种。巴氏杆菌.在16SrRNA序列和ERIC-PCR分析之后,卵黄S.亚种表现出相同的簇和带谱,可以很容易地彼此区分并进行克隆鉴定。氟苯尼考的敏感性高,青霉素,利福平,在分离物中检测到万古霉素,不管亚种的多样性。两个亚种对杆菌肽都表现出很高的抗性,克林霉素,多西环素,四环素,甲氧苄啶-磺胺甲恶唑,和红霉素和多种MDR谱,表明它们有可能成为超级细菌.来自Türkiye的第一份报告证明了鹅中gallolyticus亚种的出现。鉴于最近在蒂尔基耶的鹅产量和鹅肉消费量的增加,不应忽视鹅的发生,以防止人畜共患传播。
    Recently, an increased number of reports have described pathogens of animal origin that cause a variety of infections and a rise in their transmission to humans. Streptococcus gallolyticus, a member of the Streptococcus bovis/Streptococcus equinus complex (SBSEC), is one of these pathogens and infects a wide range of hosts from mammals to poultry and has a broad functionality ranging from pathogenicity to food fermentation. As S. gallolyticus causes complications including bacteremia, infective endocarditis, and colorectal malignancy in humans, it is important to investigate its occurrence in various hosts, including geese, to prevent potential zoonotic transmissions. This study aimed to investigate the presence of S. gallolyticus in the droppings of clinically healthy and diarrheic geese, which were raised intensively and semi-intensively, by the in vitro culture method, characterize the isolates recovered by PCR and sequence-based molecular methods and determine their antibiotic susceptibility by the disk diffusion and gradient test methods. For this purpose, 150 samples of fresh goose droppings were used. Culture positivity for S. gallolyticus was determined as 8% (12/150). PCR analysis identified 54.55% (n = 6) of the isolates as S. gallolyticus subsp. gallolyticus and 45.45% (n = 5) as S. gallolyticus subsp. pasteurianus. Following the 16S rRNA sequence and ERIC-PCR analyses, S. gallolyticus subspecies exhibited identical cluster and band profiles that could be easily distinguished from each other and were clonally identified. High rates of susceptibility to florfenicol, penicillin, rifampicin, and vancomycin were detected among the isolates, regardless of the subspecies diversity. Both subspecies showed high levels of resistance to bacitracin, clindamycin, doxycycline, tetracycline, trimethoprim-sulfamethoxazole, and erythromycin and multiple MDR profiles, indicating their potential to become superbugs. This first report from Türkiye demonstrates the occurrence of the S. gallolyticus subspecies in geese. In view of the recent increase of geese production and the consumption of goose meat in Türkiye, the occurrence of S. gallolyticus in geese should not be ignored to prevent zoonotic transmission.
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  • 文章类型: Journal Article
    目的:肠道菌群与结直肠癌的发生、发展密切相关。然而,肿瘤组织(TT)和正常组织(NT)之间的细菌共丰度组(CAGs)的差异,以及它们与临床特征的关联,需要澄清。
    方法:使用251例结直肠癌患者的TT样本和NT样本进行细菌16SrRNA测序。微生物多样性,分类学特征,微生物组成,比较TT和NT的功能通路。分层聚类用于构建CAGs。
    结果:在层次聚类分析中对四个CAG进行分组。CAG2,主要由病原菌组成,在TT样品中显著富集(TT中2.27%与NT中的0.78%,p<0.0001)。CAG4主要由非致病菌组成,在NT样品中显著富集(TT与NT中的0.79%,p=0.0004)。此外,CAG2也与肿瘤微卫星不稳定性显着相关(不稳定与不稳定的13.2%稳定2.0%,p=0.016),CAG4与CA199水平呈正相关(r=0.17,p=0.009)。
    结论:我们的研究将加深我们对多种细菌之间相互作用的理解,并提供对NT向TT过渡的潜在机制的见解。
    OBJECTIVE: Gut microbiota is closely related to the occurrence and development of colorectal cancer (CRC). However, the differences in bacterial co-abundance groups (CAGs) between tumor tissue (TT) and normal tissue (NT), as well as their associations with clinical features, are needed to be clarified.
    METHODS: Bacterial 16 S rRNA sequencing was performed by using TT samples and NT samples of 251 patients with colorectal cancer. Microbial diversity, taxonomic characteristics, microbial composition, and functional pathways were compared between TT and NT. Hierarchical clustering was used to construct CAGs.
    RESULTS: Four CAGs were grouped in the hierarchical cluster analysis. CAG 2, which was mainly comprised of pathogenic bacteria, was significantly enriched in TT samples (2.27% in TT vs. 0.78% in NT, p < 0.0001). CAG 4, which was mainly comprised of non-pathogenic bacteria, was significantly enriched in NT samples (0.62% in TT vs. 0.79% in NT, p = 0.0004). In addition, CAG 2 was also significantly associated with tumor microsatellite instability (13.2% in unstable vs. 2.0% in stable, p = 0.016), and CAG 4 was positively correlated with the level of CA199 (r = 0.17, p = 0.009).
    CONCLUSIONS: Our research will deepen our understanding of the interactions among multiple bacteria and offer insights into the potential mechanism of NT to TT transition.
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  • 文章类型: Journal Article
    阿尔及利亚东北部拥有众多温泉,然而,这些热液场所在很大程度上仍未探索其微生物生态。本研究探讨了两个不同的阿尔及利亚温泉(HammamamSaida和HammamDebagh)中的细菌丰度和多样性,并研究了盛行细菌与地球化学参数之间的联系。水和沉积物样品的高通量16SrRNA基因测序显示,在两个春季,细菌的优势均为99.85-91.16%,而古菌(0.14-0.66%)。有趣的是,赛达温泉,具有较高的温度和钠含量,藏有一个由Pseudomonadota(51.13%)主导的社区,而Debagh,Ca-Cl-SO4型弹簧,主要由芽孢杆菌居住,占55.33%。拟杆菌在两个站点上的分布均匀。额外的门,包括氯氟科,Deinococcota,蓝细菌,和叶绿素,也在场。环境因素,特别是温度,钠,钾,和碱度,显著影响细菌多样性和组成。这些发现揭示了不同微生物群落及其相关地球化学性质之间的相互作用,为未来在不同环境条件驱动的这些独特生态系统中的生物地球化学过程的研究提供有价值的见解,包括在生物修复和酶发现中的潜在应用。
    Northeastern Algeria boasts numerous hot springs, yet these hydrothermal sites remain largely unexplored for their microbial ecology. The present study explores the bacterial abundance and diversity within two distinct Algerian hot springs (Hammam Saïda and Hammam Debagh) and investigates the link between the prevailing bacteria with geochemical parameters. High-throughput 16S rRNA gene sequencing of water and sediment samples revealed a bacterial dominance of 99.85-91.16% compared to Archaea (0.14-0.66%) in both springs. Interestingly, Saïda hot spring, characterized by higher temperatures and sodium content, harbored a community dominated by Pseudomonadota (51.13%), whereas Debagh, a Ca-Cl-SO4 type spring, was primarily populated by Bacillota with 55.33%. Bacteroidota displayed even distribution across both sites. Additional phyla, including Chloroflexota, Deinococcota, Cyanobacteriota, and Chlorobiota, were also present. Environmental factors, particularly temperature, sodium, potassium, and alkalinity, significantly influenced bacterial diversity and composition. These findings shed light on the interplay between distinct microbial communities and their associated geochemical properties, providing valuable insights for future research on biogeochemical processes in these unique ecosystems driven by distinct environmental conditions, including potential applications in bioremediation and enzyme discovery.
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  • 文章类型: Journal Article
    探讨中国老年心血管疾病(CVD)患者肠道菌群的变化及其与三甲胺(TMA)代谢的相关性。使用16SrRNA测序分析了患有CVD的老年人和健康老年人的肠道菌群组成,使用顶空气相色谱(HS-GC)检测老年人粪便中的TMA水平,使用实时荧光定量聚合酶链反应(qPCR)对老年人中四种具有特征性的产TMA肠道细菌进行定量。结果表明,Firmicutes,放线菌,变形杆菌,拟杆菌,Verrucomicbia是中国老年人肠道菌群的优势微生物。健康老年人和心血管疾病患者的肠道细菌组成存在显着差异,伴随着TMA含量的显着差异。心血管疾病老年人肠道菌群的丰富度和多样性高于健康老年人。相关分析表明,某些明显不同的肠道菌群与TMA水平相关。我们的发现表明,健康老年人和CVD患者之间产生TMA的肠道菌群存在显着差异。发现TMA水平与肺炎克雷伯菌呈正相关且显着相关,这表明这种细菌与老年人肠道中TMA的产生密切相关。这可能对老年人群中CVD的发展和进展有影响。
    To investigate the changes in the intestinal flora in the Chinese elderly with cardiovascular disease (CVD) and its correlation with the metabolism of trimethylamine (TMA), the intestinal flora composition of elderly individuals with CVD and healthy elderly individuals was analyzed using 16S rRNA sequencing, the TMA levels in the feces of elderly were detected using headspace-gas chromatography (HS-GC), and four kinds of characterized TMA-producing intestinal bacteria in the elderly were quantified using real-time fluorescence quantitative polymerase chain reaction (qPCR). The results showed that Firmicutes, Actinobacteria, Proteobacteria, Bacteroidetes, and Verrucomicrobia are the dominant microorganisms of the intestinal flora in the Chinese elderly. And there were significant differences in the intestinal bacteria composition between healthy elderly individuals and those with CVD, accompanied by a notable difference in the TMA content. The richness and diversity of the intestinal flora in the elderly with CVD were higher than those in the healthy elderly. Correlation analysis indicated that certain significantly different intestinal flora were associated with the TMA levels. Our findings showed a significant difference in TMA-producing intestinal flora between healthy elderly individuals and those with CVD. The TMA levels were found to be positively and significantly correlated with Klebsiella pneumoniae, suggesting that this bacterium is closely linked to the production of TMA in the elderly gut. This may have implications for the development and progression of CVD in the elderly population.
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  • 文章类型: Journal Article
    这项研究使用了0日龄(36只兔子)的粪便,10天大(119只兔子),和60日龄(119只)子代兔及其相应的雌性兔(36只)为实验材料。使用16srRNA测序,研究分析了不同生长发育阶段家兔肠道菌群的类型和变化,以及肠道菌群组成与60日龄兔体重的相关性。将所有实验兔置于同一兔棚内。幼年兔在18日龄时饲喂固体饲料,在35日龄时断奶。除了确定不同年龄阶段兔肠道微生物群的优势细菌门,研究发现,所有乳兔(10日龄)中的梭状芽胞杆菌和副营养梭状芽胞杆菌的丰度均显着高于全饲料(60日龄)(p<0.05),革兰阴性菌cTPY13的丰度显著降低(p<0.05)。此外,快速预期最大微生物来源追踪(FEAST)分析表明,雌性兔肠道菌群对后代兔肠道菌群定植的贡献显着高于无关兔肠道菌群(p<0.05)。雌性兔肠道菌群对0日龄兔肠道菌群定植的贡献显著高于10日龄和60日龄兔肠道菌群定植的贡献(p<0.05)。最后,基于混合线性模型分析60日龄兔肠道菌群组成与体重的相关性,筛选了6个显著影响体重的ASV。上述结果为维护肠道健康提供了重要的理论和实践指导,提高增长和发展绩效,和兔子的喂养配方。
    This study used feces from 0-day-old (36 rabbits), 10-day-old (119 rabbits), and 60-day-old (119 rabbits) offspring rabbits and their corresponding female rabbits (36 rabbits) as experimental materials. Using 16s rRNA sequencing, the study analyzed the types and changes of gut microbiota in rabbits at different growth and development stages, as well as the correlation between gut microbiota composition and the weight of 60-day-old rabbits. All experimental rabbits were placed in the same rabbit shed. Juvenile rabbits were fed solid feed at 18 days of age and weaned at 35 days of age. In addition to identifying the dominant bacterial phyla of gut microbiota in rabbits at different age stages, it was found that the abundance of Clostridium tertium and Clostridium paraputrificum in all suckling rabbits (10-day-old) was significantly higher than that in rabbits fed with whole feed (60-day-old) (p < 0.05), while the abundance of Gram-negative bacterium cTPY13 was significantly lower (p < 0.05). In addition, Fast Expected Maximum Microbial Source Tracing (FEAST) analysis showed that the contribution of female rabbits\' gut microbiota to the colonization of offspring rabbits\' gut microbiota was significantly higher than that of unrelated rabbits\' gut microbiota (p < 0.05). The contribution of female rabbits\' gut microbiota to the colonization of gut microbiota in 0-day-old rabbits was significantly higher than that to the colonization of gut microbiota in the 10- and 60-day-old rabbits (p < 0.05). Finally, the correlation between gut microbiota composition and body weight of 60-day-old rabbits was analyzed based on a mixed linear model, and six ASVs significantly affecting body weight were screened. The above results provide important theoretical and practical guidance for maintaining gut health, improving growth and development performance, and feeding formulation in rabbits.
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  • 文章类型: Journal Article
    16S核糖体RNA(rRNA)基因的高变区测序通过提供对特定生态位内细菌群落的见解,在微生物生态学中起着至关重要的作用。在提供有价值的属级信息的同时,它对目标遗传区域数据的依赖限制了它的整体效用。测序技术的最新进展已经能够表征全长16SrRNA基因,加强物种级分类。尽管当前的短读取平台具有成本效益且精确,它们缺乏全长16SrRNA扩增子测序能力。这项研究旨在通过利用标准模拟微生物群落并随后进行探索性表征,评估在IlluminaiSeq100和16SrRNA扩增子装配工作流程上修改的150bp配对末端全长16SrRNA扩增子短读取测序技术的可行性圈养(动物园)和自由放养的非洲象(Loxodontaafricana)呼吸微生物群。我们的研究结果表明,尽管产生了组装的扩增子,平均长度为869bp,这种测序技术提供了与其他哺乳动物的模拟群落和呼吸道微生物群的理论组成一致的分类分配。初步细菌特征,视觉识别可能代表不同的呼吸道隔室(躯干和下呼吸道),有必要进行进一步的调查,以更深入地了解它们对大象生理和健康的影响。
    Hypervariable region sequencing of the 16S ribosomal RNA (rRNA) gene plays a critical role in microbial ecology by offering insights into bacterial communities within specific niches. While providing valuable genus-level information, its reliance on data from targeted genetic regions limits its overall utility. Recent advances in sequencing technologies have enabled characterisation of the full-length 16S rRNA gene, enhancing species-level classification. Although current short-read platforms are cost-effective and precise, they lack full-length 16S rRNA amplicon sequencing capability. This study aimed to evaluate the feasibility of a modified 150 bp paired-end full-length 16S rRNA amplicon short-read sequencing technique on the Illumina iSeq 100 and 16S rRNA amplicon assembly workflow by utilising a standard mock microbial community and subsequently performing exploratory characterisation of captive (zoo) and free-ranging African elephant (Loxodonta africana) respiratory microbiota. Our findings demonstrate that, despite generating assembled amplicons averaging 869 bp in length, this sequencing technique provides taxonomic assignments consistent with the theoretical composition of the mock community and respiratory microbiota of other mammals. Tentative bacterial signatures, potentially representing distinct respiratory tract compartments (trunk and lower respiratory tract) were visually identified, necessitating further investigation to gain deeper insights into their implication for elephant physiology and health.
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