16s rRNA sequencing

16S rRNA 测序
  • 文章类型: Journal Article
    最近对帕金森病(PD)的见解,进行性神经退行性疾病,提示肠道微生物组对其发病机理和通过肠-脑轴的进展有重大影响。本研究整合了16SrRNA测序,高通量转录组测序,和动物模型实验,以探索支持肠-脑轴在PD中的作用的分子机制,关注由SCFA受体FFAR2和FFAR3介导的短链脂肪酸(SCFA)。我们的发现强调了PD患者和健康个体之间肠道菌群组成的显著差异,特别是在类群中,如大肠杆菌志贺氏菌和拟杆菌,这可能通过次级代谢产物生物合成影响SCFA水平。值得注意的是,从健康的粪便微生物群移植(FMT)到1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的PD小鼠模型显着改善运动功能,纹状体中多巴胺和5-羟色胺水平增强,并增加黑质中多巴胺能神经元的数量,同时减少神经胶质细胞的活化。这种治疗效果与SCFA水平的增加有关,如乙酸盐,丙酸盐,MPTP损伤小鼠肠道中的丁酸盐。此外,转录组学分析显示MPTP损伤小鼠中FFAR2和FFAR3的表达上调,表明它们在介导FMT对中枢神经系统的益处中的关键作用。这些结果提供了令人信服的证据,表明肠道微生物群和SCFA在调节肠-脑轴中起着关键作用。为PD的病因和治疗干预的潜在目标提供了新的见解。
    Recent insights into Parkinson\'s disease (PD), a progressive neurodegenerative disorder, suggest a significant influence of the gut microbiome on its pathogenesis and progression through the gut-brain axis. This study integrates 16S rRNA sequencing, high-throughput transcriptomic sequencing, and animal model experiments to explore the molecular mechanisms underpinning the role of gut-brain axis in PD, with a focus on short-chain fatty acids (SCFAs) mediated by the SCFA receptors FFAR2 and FFAR3. Our findings highlighted prominent differences in the gut microbiota composition between PD patients and healthy individuals, particularly in taxa such as Escherichia_Shigella and Bacteroidetes, which potentially impact SCFA levels through secondary metabolite biosynthesis. Notably, fecal microbiota transplantation (FMT) from healthy to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse models significantly improved motor function, enhanced dopamine and serotonin levels in the striatum, and increased the number of dopaminergic neurons in the substantia nigra while reducing glial cell activation. This therapeutic effect was associated with increased levels of SCFAs such as acetate, propionate, and butyrate in the gut of MPTP-lesioned mice. Moreover, transcriptomic analyses revealed upregulated expression of FFAR2 and FFAR3 in MPTP-lesioned mice, indicating their crucial role in mediating the benefits of FMT on the central nervous system. These results provide compelling evidence that gut microbiota and SCFAs play a critical role in modulating the gut-brain axis, offering new insights into PD\'s etiology and potential targets for therapeutic intervention.
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  • 文章类型: Journal Article
    目的:饲养酒厂副产品(WBP)可能会影响牛微生物组,因为它们的酚类化合物和WBP相关微生物群的转移。这项工作检查了包含WBP后未充分开发的固体相关瘤胃微生物组的变化。
    方法:使用瘤胃模拟技术,用供体母牛的接种物接种发酵罐,并饲喂六种饮食处理之一,包括70%干草+30%浓缩物(CON)的对照饮食,对照饮食+3.7%商业葡萄籽提取物(EXT),65%干草+25%精矿+10%葡萄果渣(GP-低),56%干草+24%浓缩+20%葡萄渣(GP-high),70%干草+25%浓缩物+5%葡萄籽粉(GS-低),和65%干草+25%浓缩物+10%葡萄籽粉(GS-高)(干物质基础)。细菌的组成变化,固体部分中的古细菌和真菌基于16S和ITS2rRNA测序。
    结果:微生物群的α-和β-多样性未受影响。然而,处理改变了低分类水平的细菌组成。Butyrivibriofibrisolvens,Bryantii密螺旋体,细菌MC2010在EXT中减少,与CON相比,柏林密螺旋体的GP高和GP低增加。关于真菌,GS-高增加的念珠菌属。,Lachanceaspp.,Microdochiumspp.,Mucorspp.,Pichiaspp.,Saturnisporaspp。,和酵母菌属。与CON相比。在WBP样品中检测到许多非酵母属酵母,但在供体母牛和CON样品中却没有。受处理影响的属不是营养物质瘤胃降解的主要贡献者。
    结论:结果表明,当作为提取物递送时,瘤胃固体细菌对葡萄酚的敏感性以及WBP相关微生物群转移到瘤胃中。
    OBJECTIVE: Feeding winery by-products (WBP) could affect the bovine microbiome because of their phenol compounds and a transfer of WBP-associated microbiota. This work examined changes in the underexplored solid-associated rumen microbiome following the inclusion of WBP.
    METHODS: Using the rumen simulation technique, fermenters were inoculated with the inoculum of donor cows and were fed one of six dietary treatments including a control diet of 70 % hay +30 % concentrate (CON), control diet + 3.7 % commercial grapeseed extract (EXT), 65 % hay + 25 % concentrate + 10 % grape pomace (GP-low), 56 % hay + 24 % concentrate + 20 % grape pomace (GP-high), 70 % hay + 25 % concentrate + 5 % grapeseed meal (GS-low), and 65 % hay + 25 % concentrate + 10 % grapeseed meal (GS-high) (dry matter basis). The compositional changes of bacteria, archaea and fungi in the solid fractions were based on 16S and ITS2 rRNA sequencing.
    RESULTS: The alpha- and beta-diversity of the microbiota were unaffected. However, treatment modified the bacterial composition at low taxonomic levels. Butyrivibrio fibrisolvens, Treponema bryantii, and bacterium MC2010 decreased in EXT, while Treponema berlinense was increased in GP-high and GP-low compared to CON. Concerning fungi, GS-high increased Candida spp., Lachancea spp., Microdochium spp., Mucor spp., Pichia spp., Saturnispora spp., and Zygosaccharomyces spp. compared to CON. Many non-Saccharomyces yeasts were detected in WBP samples but absent in donor cows and CON samples. The genera affected by treatment were not the major contributors to the ruminal degradation of nutrients.
    CONCLUSIONS: The results indicate a sensitivity of rumen solid bacteria to grape phenols when delivered as an extract and a transfer of WBP-associated microbiota into the rumen.
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  • 文章类型: Journal Article
    背景:最近,口腔癌杆菌具核梭杆菌(F.核仁),与溃疡性结肠炎(UC)有关。这里,我们的目的是研究粪便微生物移植(FMT)是否可以通过恢复肠道菌群和消除口腔来源的F.核仁和毒力因子fadA来缓解UC。
    方法:C57BL/6J小鼠随机分为健康对照组,葡聚糖硫酸钠基团(DSS),口服接种组(OR),上FMT组(UFMT),和较低的FMT组(LFMT)。疾病活动指数,体重,存活率,和组织病理学评分用于测量结肠炎的严重程度。通过对紧密连接蛋白Occludin进行免疫组织化学染色来评估肠粘膜屏障的功能。使用实时PCR评估nusG基因和毒力基因fadA的相对丰度。通过ELISA检测细胞因子水平。使用16SrRNA的全长测序来分析肠道菌群的变化和组成。
    结果:核仁F.的口服孵育进一步加剧了结肠炎和肠道菌群失调的严重程度。肽链球菌科,肠球菌科,和大肠杆菌在OR小鼠中显著富集。然而,与UFMT小鼠相比,LFMT小鼠的疾病活动明显减少,在恢复肠道菌群和消除核仁F.方面更有效。拟杆菌,落叶松科,Prevotellaceae主要是LFMT小鼠的富集细菌。此外,属如乳酸菌,Allobaculum,发现拟杆菌与TNF-α呈负相关,IL-1β,IL-6像Romboutsia这样的属人,大肠杆菌志贺氏菌,肠球菌,发现梭菌与TNF-α呈正相关,IL-1β,IL-6
    结论:经口孵育核菌进一步加重了DSS诱导的结肠炎小鼠的严重程度和菌群失调。此外,下肠道FMT可以通过恢复肠道菌群多样性和消除F.核仁和毒力因子fadA来改善结肠炎。
    BACKGROUND: Recently, the oral oncobacterium Fusobacterium nucleatum (F. nucleatum), has been linked with ulcerative colitis (UC). Here, we aim to investigate whether Fecal Microbiota Transplantation (FMT) can alleviate UC by restoring gut microbiota and eliminating oral-derived F. nucleatum and virulence factor fadA.
    METHODS: C57BL/6J mice were randomly divided into a healthy control group (HC), Dextran Sulfate Sodium group (DSS), oral inoculation group (OR), upper FMT group (UFMT), and lower FMT group (LFMT). Disease activity index, body weight, survival rate, and histopathological scores were used to measure the severity of colitis. The function of the intestinal mucosal barrier was evaluated by performing immunohistochemical staining of the tight junction protein Occludin. Real-time PCR was used to assess the relative abundance of the nusG gene and the virulence gene fadA. Cytokine levels were detected by ELISA. Full-length sequencing of 16S rRNA was used to analyze the changes and composition of gut microbiota.
    RESULTS: Oral incubation of F. nucleatum further exacerbated the severity of colitis and gut dysbiosis. Peptostreptococcaceae, Enterococcaceae, and Escherichia coli were significantly enriched in OR mice. However, LFMT mice showed an obvious decrease in disease activity and were more effective in restoring gut microbiota and eliminating F. nucleatum than UFMT mice. Bacteroidota, Lachnospiraceae, and Prevotellaceae were mainly enriched bacteria in LFMT mice. In addition, Genera such as Lactobacillus, Allobaculum, and Bacteroidales were found negative correlation with TNF-α, IL-1β, and IL-6. Genera like Romboutsia, Escherichia Shigella, Enterococcus, and Clostridium were found positively correlated with TNF-α, IL-1β, and IL-6.
    CONCLUSIONS: Oral incubation of F. nucleatum further exacerbates the severity and dysbiosis in DSS-induced colitis mice. Besides, lower tract FMT can ameliorate colitis by restoring the gut microbiota diversity and eliminating F. nucleatum and virulence factor fadA.
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  • 文章类型: Journal Article
    全面了解口腔微生物组在中度至重度斑块型银屑病中的作用及其对疾病管理和发展的潜在影响具有重要意义。为了探讨口腔微生物群与重度银屑病的相关性,这项研究涉及72名严重银屑病患者和16名健康个体,仔细记录其临床表现和生活习惯。采用16SrRNA基因测序和生物信息学分析等前沿技术对微生物菌群进行比较,研究严重斑块型银屑病患者的动态变化,银屑病关节炎患者和健康个体。研究结果揭示了值得注意的模式,包括银屑病关节炎组的Aggregatibacter水平升高,伴随着普雷沃氏菌水平的下降。此外,Capnocytandophaga的富集(P=0.009),弯曲杆菌(P=0.0022),和醋杆菌(P=0.0292)在银屑病组中明显高于对照组,而某些细菌物种,如拟杆菌(P=0.0049),Muribaculaceae(P=0.0048)显示出减少的富集。此外,银屑病关节炎组表现出明显更高水平的Ralstonia,双歧杆菌和小单孢菌。基于这些发现,可以推断,普雷沃氏菌水平较低,棒状杆菌水平较高的个体可能更容易发生银屑病加重.
    Gaining a comprehensive understanding of the role played by the oral microbiome in moderate to severe plaque psoriasis and its potential implications for disease management and development holds significant importance. With the objective of exploring correlations between the oral microbiota and severe psoriasis, this study involved 72 severe psoriasis patients and 16 healthy individuals, whose clinical manifestations and living habits were carefully recorded. Cutting-edge techniques such as 16S rRNA gene sequencing and bioinformatics analysis were employed to compare the microbial flora, investigating dynamic changes among severe plaque psoriasis patients, psoriatic arthritis patients and healthy individuals. The findings revealed noteworthy patterns including increased levels of Aggregatibacter in the psoriatic arthritis group, accompanied by a decrease in the level of Prevotella. Moreover, the enrichment o Capnocytandophaga (P = 0.009), Campylobacter (P = 0.0022), and Acetobacter (P = 0.0292) was notably more substantial in the psoriasis group compared to the control group, whereas certain bacterial species such as Bacteroides (P = 0.0049), Muribaculaceae (P = 0.0048) demonstrated decreased enrichment. Additionally, the psoriatic arthritis group exhibited significantly higher levels of Ralstonia, Bifidobacterium and Micromonospora. Based on these findings, it can be inferred that individuals with lower levels of Prevotella and higher levels of Corynebacterium may be more susceptible to psoriasis exacerbation.
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  • 文章类型: Journal Article
    目的采用基于微生物培养的方法,对健康孕妇和感染HPV的孕妇阴道微生物优势群的组成进行比较分析。MALDITOFMS方法和16SrRNA基因片段测序用于鉴定从健康孕妇(n=32)和感染HPV的孕妇(n=24)中分离的微生物。发现两组的阴道分泌物样品都含有4种细菌:芽孢杆菌,放线菌,Pseudomonadota,拟杆菌,和子囊真菌。健康孕妇中最常见的微生物群落是CSTI(p=0.0007),和CSTV在感染HPV的孕妇中(p=0.0001)。在属一级,在所有样本中共发现25个分类群,乳杆菌是总体上的优势属。在HPV感染的患者中,埃希氏菌(p<0.0001)和普雷沃氏菌(p=0.0001)的浓度较高。计算门的皮尔逊相关系数时,发现芽孢杆菌与HPV基因型16和51呈负相关(p≤0.05),但HPV基因型为59(p≤0.05),就像放线菌一样(p≤0.05)。拟杆菌与HPV基因型56呈正相关(0.001 The purpose of this study was to conduct a comparative analysis of the composition of the dominant groups of vaginal microorganisms in healthy pregnant women and pregnant women infected with HPV using a microbiological culture-based method. The MALDI TOF MS method and 16S rRNA gene fragment sequencing were used to identify microorganisms isolated from healthy pregnant women (n=32) and pregnant women infected with HPV (n=24). It was found that vaginal secretion samples from both groups contained bacteria of 4 phyla: Bacillota, Actinomycetota, Pseudomonadota, Bacteroidota, and Ascomycota fungi. The most common microbial community in healthy pregnant women being CST I (p=0.0007), and CST V in pregnant women infected with HPV (p=0.0001). At the genus level, a total of 25 taxa were found in all samples, with Lactobacillus being the dominant genus overall. Escherichia (p<0.0001) and Prevotella (p=0.0001) concentrations were higher in HPV infected patients. When calculating the Pearson correlation coefficient for the phyla, it was found that Bacillota correlated negatively with HPV genotypes 16 and 51 (p≤0.05), but positively with HPV genotype 59 (p≤0.05), just like Actinomycetota (p≤0.05). Bacteroidota correlated positively with HPV genotype 56 (0.001
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  • 文章类型: Journal Article
    妇科检查期间常规使用凝胶润滑,以防止或减轻疼痛,但其对采样过程中微生物组成的影响尚不清楚.本研究旨在研究润滑凝胶是否会影响阴道样品的微生物组成。我们包括31名孕妇,在他们的第三个三个月期间出现在诊所或急诊室,并收集了143个独特的阴道样本进行16S扩增子微生物分析。在各种条件下使用无菌拭子获得阴道样品:无凝胶立即冷冻(n=30),用凝胶立即冷冻,没有凝胶-在室温(RT)冷冻前5小时,在冷冻前在室温下使用凝胶5小时,和24小时后没有凝胶立即冷冻的额外取样。我们发现,采用凝胶润滑的样品收集会影响样品质量-与没有凝胶的样品相比,一半的凝胶样品未能达到加工限制。凝胶对测试质量的影响在24小时后消失。然而,当样品遇到测序后过滤器时,凝胶润滑不会改变微生物组成,个体分类群丰度或α和β多样性。我们建议在凝胶暴露前或24小时后取样。这些发现强调了在阴道微生物组研究中考虑样品收集方法的重要性,以确保高质量的微生物数据进行准确分析。
    Gel lubrication is routinely used during gynecological examination to prevent or reduce pain, yet its impact on microbial composition during sampling remains unclear. This study aimed to investigate whether lubricating gel affects the microbial composition of vaginal samples. We included 31 pregnant women presenting during their third trimester to clinics or emergency room and collected 143 unique vaginal samples for 16S amplicon microbial analysis. Vaginal samples were obtained using sterile swabs under various conditions: without gel-immediately frozen (n = 30), with gel-immediately frozen, without gel-at room temperature (RT) for 5 h before freezing, with gel-at RT for 5 h before freezing, and additional sampling after 24 h without gel-immediate freezing. We found that sample collection with gel lubrication influenced specimen quality-half of the gel samples failing to meet processing limitation compared to those without gel. The effect of gel on testing quality dissipated after 24 h. However, when samples met post-sequencing filters, gel lubrication did not alter the microbial composition, individual taxa abundance or alpha and beta diversity. We recommend sampling either before gel exposure or 24 h after. These findings underscore the importance of considering sample collection methodologies in vaginal microbiome studies to ensure high-quality microbial data for accurate analysis.
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  • 文章类型: Journal Article
    本研究旨在评估扩展的基质辅助激光解吸电离飞行时间质谱(MALDI-TOFMS)数据库,以鉴定流感嗜血杆菌(Hi)以外的嗜血杆菌。
    总共144种嗜血杆菌,从囊性纤维化患者(生活)的呼吸道样本中培养出来,用MALDI-TOFMS和16SrRNA测序鉴定。其中,99个嗜血杆菌菌株显示与国家生物技术信息中心(NCBI)数据库中的最佳匹配菌株>99%的相似性,并且使用MALDI-TOFMS和16SrRNA测序将其分配到单个嗜血杆菌亚种。将菌株子集(n=58/99)的MS谱添加到BrukerMALDI-TOFMS数据库中。随后,重新分析先前在常规设置中分析的270种不同菌株。
    16SrRNA测序可靠地鉴定了99/144个嗜血杆菌菌株(>99%相似性)。H.溶血杆菌16SrRNA鉴定是次优的,因为只有3/21H.溶血杆菌菌株与NCBI数据库中的溶血杆菌16SrRNA序列达到>99%的相似性。MALDI-TOFMS数据库的扩展仅适度地改善了溶血嗜血杆菌的可靠识别数量,流感嗜血杆菌和副嗜血杆菌(<10%)。相比之下,对副溶血H.H.副流感,H.putorum和H.pittmaniae(>85%)。
    16SrRNA测序是鉴定嗜血杆菌的一种有价值的方法。除了嗨。MALDI-TOFMS数据库的扩展,基于16SrRNA测序结果,增加了可靠鉴定的比例,在这项研究中,嗜血杆菌增加了10%。除了Hi菌株识别。
    UNASSIGNED: This study aimed to evaluate an expanded matrix-assisted laser desorption-ionization-time of flight mass spectrometry (MALDI-TOF MS) database for the identification of Haemophilus species other than H. influenzae (Hi).
    UNASSIGNED: A total of 144 Haemophilus species, cultured from respiratory samples from people (living) with cystic fibrosis, were identified with MALDI-TOF MS and 16S rRNA sequencing. Of these, 99 Haemophilus strains showed >99% similarity with the best matching strain in the National Center for Biotechnology Information (NCBI) database and were assigned to a single Haemophilus subspecies using both MALDI-TOF MS and 16S rRNA sequencing. The MS profiles of a subset of strains (n = 58/99) were added to the Bruker MALDI-TOF MS database. Subsequently, 270 different strains that were analyzed previously in a routine setting were re-analyzed.
    UNASSIGNED: 16S rRNA sequencing reliably identified 99/144 Haemophilus strains (>99% similarity). H. haemolyticus 16S rRNA identification was suboptimal since only 3/21 H. haemolyticus strains attained a similarity of >99% with H. haemolyticus 16S rRNA sequence in the NCBI database. Expansion of the MALDI-TOF MS database improved the number of reliable identifications only moderately for H. haemolyticus, H. influenzae and H. paraphrohaemolyticus (<10%). By contrast, improved identification was more outspoken for H. parahaemolyticus, H. parainfluenzae, H. sputorum and H. pittmaniae (>85%).
    UNASSIGNED: 16S rRNA sequencing is a valuable method for the identification of Haemophilus sp. other than Hi. Expansion of the MALDI-TOF MS database, based on 16S rRNA sequencing results, increased the proportion of reliable identifications and in this study resulted in an increase of 10% of Haemophilus sp. other than Hi strain identifications.
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  • 文章类型: Journal Article
    宿主-微生物群相互作用在宿主生物学中起着关键作用,因此,阐明形成肠道微生物群落的机制至关重要。我们利用三松stickleback鱼(Gasterosteusaculeatus)作为模型系统,以研究宿主和环境因素对肠道微生物群变化的贡献。这些鱼提供了在自然条件下进行实验的独特机会;我们在实验池塘中从三个不同的湖泊中饲养了底栖和边缘生态型,允许我们评估共享环境(池塘)的相对影响,地理起源(起源湖),营养生态学和遗传学(生态型)和生物学性别对肠道微生物群α-和β-多样性的影响。宿主生态型对α-多样性的影响最强,底栖鱼类表现出比海洋鱼类更高的多样性,其次是饲养环境。β-多样性主要是由饲养环境决定的,其次是宿主生态型,这表明环境因素在决定肠道菌群组成中起着至关重要的作用。此外,许多细菌在池塘中的数量不同,强调环境因素对肠道微生物群变异的重大贡献。我们的研究说明了环境与宿主生态或遗传因素之间的复杂相互作用,在塑造棘鱼肠道微生物群中,并强调了在自然条件下进行的实验对理解肠道微生物群动态的价值。
    Host-microbiota interactions play a critical role in the hosts\' biology, and thus, it is crucial to elucidate the mechanisms that shape gut microbial communities. We leveraged threespine stickleback fish (Gasterosteus aculeatus) as a model system to investigate the contribution of host and environmental factors to gut microbiota variation. These fish offer a unique opportunity for experiments in naturalistic conditions; we reared benthic and limnetic ecotypes from three different lakes in experimental ponds, allowing us to assess the relative effects of shared environment (pond), geographic origin (lake-of-origin), trophic ecology and genetics (ecotype) and biological sex on gut microbiota α- and β-diversity. Host ecotype had the strongest influence on α-diversity, with benthic fish exhibiting higher diversity than limnetic fish, followed by the rearing environment. β-diversity was primarily shaped by rearing environment, followed by host ecotype, indicating that environmental factors play a crucial role in determining gut microbiota composition. Furthermore, numerous bacterial orders were differentially abundant across ponds, underlining the substantial contribution of environmental factors to gut microbiota variation. Our study illustrates the complex interplay between environmental and host ecological or genetic factors in shaping the stickleback gut microbiota and highlights the value of experiments conducted under naturalistic conditions for understanding gut microbiota dynamics.
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  • 文章类型: Journal Article
    先前观察到对精子质量参数的季节性影响。尽管以前通过16SrRNA测序鉴定了公牛精液微生物群,它没有在不同季节的商业精液样本中进行,其与精子质量参数的关系尚未得到评估。这项研究的目的是:(i)评估不同季节公牛精液微生物群和精子质量参数的多样性,和(ii)发现特定细菌是否与特定精子质量参数的季节性差异有关。在3个季节的54个商业公牛精液样本中鉴定出公牛精液微生物群(冬季,spring,summer).通过计算机辅助精子分析(CASA)和流式细胞术(FC)分析精子质量。所有样本中的28个门,在所有季节的样本中都发现了六个门,观察到它们分布的季节性差异。在属一级,鉴定出388属,其中22属的相对丰度超过1%,细菌多样性表现出季节性差异,所有季节都有9个细菌属。活过氧化氢阳性精子细胞在春季和夏季之间观察到差异(P<0.05)。对于一些CASA运动学(VCL和LIN)和FC参数(高呼吸活动,和季节之间的活过氧化氢阳性精子细胞)。然而,在春季观察到精子质量参数与特定细菌之间的关联。
    A seasonal effect on sperm quality parameters was observed previously. Although identification of the bull semen microbiota by 16S rRNA sequencing was performed previously, it has not been carried out in commercial semen samples from different seasons, and its connection with sperm quality parameters has not been evaluated yet. The objectives in this study were; (i) to evaluate diversity of bull semen microbiota and sperm quality parameters in different seasons, and (ii) to find if specific bacteria were associated with seasonal differences in specific sperm quality parameters. Bull semen microbiota was identified in 54 commercial bull semen samples from 3 seasons (winter, spring, summer). Sperm quality was analysed by Computer Assisted Sperm Analyses (CASA) and Flow Cytometry (FC). From 28 phyla in all samples, six phyla were identified in samples from all seasons, with observed seasonal differences in their distribution. At genus level, 388 genera were identified, of which 22 genera had a relative abundance over 1 % and showed seasonal differences in bacterial diversity, and 9 bacteria genera were present in all seasons. Differences between spring and summer (P < 0.05) were observed for live hydrogen peroxide positive sperm cells. A trend towards significance (0.10 > P > 0.05) was observed for some CASA kinematics (VCL and LIN) and FC parameters (High respiratory activity, and live hydrogen peroxide positive sperm cells) between seasons. Nevertheless, associations between sperm quality parameters and specific bacteria were observed in spring.
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  • 文章类型: Journal Article
    根据全球统计数据,脓毒症每年造成5人死亡。在最近的研究中,脓毒症发病率至少占所有ICU入院的35%,并且死亡率很高。尤其是在并存的情况下。挑战是准确诊断致病生物,考虑到可能的多微生物感染等因素,共生和环境污染物。传统技术,如文化,自动化培养系统或甚至更新的物种特异性PCR或胶片阵列这些挑战难以克服。Bactfast®和Fungifast®分析以及集成的工作流程基于下一代测序,能够区分感染病原体的污染和共生。查明确切病原体的独特能力,考虑到各种样品的共生和污染,具有极高的灵敏度,由于其全面的覆盖范围和速度,可能使其成为非解决ICU脓毒症的诊断选择工具.这项研究的目的是评估Bactfast®和Fungifast®作为ICU环境中最后一英里诊断工具的使用。
    这项研究是考虑进入四个重症监护病房(ICU)进行的。遗留测试,主要是在文化方面,是在重症监护病房所在医院的各种综合微生物设施进行的,在钦奈,印度。NABL认可的实验室MicroGenomics(India)PvtLtd,被建立为下一代测序的中央处理设施,以运行Bactfast®和Fungifast®测定。由微生物学家组成的多学科顾问团队对490个样本的结果进行了回顾性分析,和传染病医生。
    使用Bactfast®测定法建立的诊断工作流程提供了94.1%的灵敏度和86.6%的特异性。与2017年发布的数据相比,Bactfast®中的病原体鉴定更好,这通过与临床确认的正相关反映出来。尽管Fungifast®特异性很高,99.4%,490个样本中只有12个样本对真菌培养呈阳性。因此,结论是,需要对基于多种技术的真菌进行进一步研究,并需要更多的真实阳性样本来评估测试。
    Bactfast®可以在没有任何偏差的情况下鉴定样品中的病原体。将其作为危及生命的ICU败血症的诊断方式引入可以降低死亡率和发病率。虽然Fungifast®的初步结果令人鼓舞,但需要进一步的研究才能获得更多关于测试灵敏度的信息。
    UNASSIGNED: Sepsis claims 1 in 5 lives annually as per global statistics. Sepsis incidence in recent studies represents at least 35 % of all ICU admissions and has a high mortality rate, especially in the presence of co-existing morbidities. The challenge has been to accurately diagnose the causative organism, considering factors such as possible polymicrobial infections, commensals and environmental contaminants. Legacy techniques such as culture, automated culture systems or even newer species-specific PCR or film array these challenges difficult to overcome. The Bactfast® and Fungifast® assays along with the integrated workflow is based on next generation sequencing and have the ability to demarcate infecting pathogen from contamination and commensal. The unique ability to pinpoint the exact pathogen, considering the commensal and contamination in a variety of samples, with an extremely high sensitivity could lead it to be a tool of diagnostic choice for non-resolving ICU sepsis due to its comprehensive coverage and speed. The aim of this study was to evaluate the use of Bactfast® and Fungifast® as a last mile diagnostic tool in a ICU setting.
    UNASSIGNED: This study was carried out considering access to four intensive care units (ICU). Legacy testing, mostly done on culture, was conducted at the various integrated microbiology facilities of the hospitals where the ICUs were located, in Chennai, India. NABL accredited laboratory Micro Genomics (India) Pvt Ltd, was established as the central processing facility for next generation sequencing to run the Bactfast® and Fungifast® assay. Co-relation of results for 490 samples was done retrospectively by a multi-disciplinary team of consultants which comprised of microbiologists, and infectious disease physicians.
    UNASSIGNED: The diagnostic workflow established with the Bactfast® assay provided a sensitivity of 94.1 % and specificity of 86.6 %. Identification of pathogens in Bactfast® was better when compared to the data published in 2017, as reflected by positive co-relation with clinical confirmation. Although the Fungifast® specificity was high, at 99.4 %, only 12 samples were positive on fungal culture out of 490 samples. Therefore, it was concluded a further study for fungi based on multiple technologies with more true positive samples is required to evaluate the test.
    UNASSIGNED: Bactfast® can identify pathogens in a sample without any bias. Its introduction as diagnostic modality in life threatening ICU sepsis could reduce mortality and morbidity. Although the initial results of Fungifast® are encouraging a further research is required for more information on test sensitivity.
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