Association between childhood obesity and gut microbiota:16S rRNA gene sequencing-based cohort study

BACKGROUND This study aimed to identify characteristic gut genera in obese and normal-weight children(8-12 years old)using 16S rDNA sequencing.The research aimed to provide insights for mechanistic studies and prevention strategies for childhood obesity.Thirty normal-weight and thirty age-and sex-matched obese children were included.Questionnaires and body measurements were collected,and fecal samples underwent 16S rDNA sequencing.Significant differences in body mass index(BMI)and body-fat percentage were observed between the groups.Analysis of gut microbiota diversity revealed lowerα-diversity in obese children.Differences in gut microbiota composition were found between the two groups.Prevotella and Firmicutes were more abundant in the obese group,while Bacteroides and Sanguibacteroides were more prevalent in the control group.AIM To identify the characteristic gut genera in obese and normal-weight children(8-12-year-old)using 16S rDNA sequencing,and provide a basis for subsequent mechanistic studies and prevention strategies for childhood obesity.METHODS Thirty each normal-weight,1:1 matched for age and sex,and obese children,with an obese status from 2020 to 2022,were included in the control and obese groups,respectively.Basic information was collected through questionnaires and body measurements were obtained from both obese and normal-weight children.Fecal samples were collected from both groups and subjected to 16S rDNA sequencing using an Illumina MiSeq sequencing platform for gut microbiota diversity analysis.RESULTS Significant differences in BMI and body-fat percentage were observed between the two groups.The Ace and Chao1 indices were significantly lower in the obese group than those in the control group,whereas differences were not significant in the Shannon and Simpson indices.Kruskal-Wallis tests indicated significant differences in unweighted and weighted UniFrac distances between the gut microbiota of normal-weight and obese children(P<0.01),suggesting substantial disparities in both the species and quantity of gut microbiota between the two groups.Prevotella,Firmicutes,Bacteroides,and Sanguibacteroides were more abundant in the obese and control groups,respectively.Heatmap results demonstrated significant differences in the gut microbiota composition between obese and normal-weight children.CONCLUSION Obese children exhibited lowerα-diversity in their gut microbiota than did the normal-weight children.Significant differences were observed in the composition of gut microbiota between obese and normal-weight children.

基于16S rDNA测序分析中药复方石苦秦散对腹泻小鼠盲肠的影响

【目的】探明止泻中药复方石苦秦散对蓖麻油和番泻叶所致腹泻小鼠肠道微生物的影响,为研究石苦秦散止泻作用机制奠定基础。【方法】将70只小鼠分为正常对照组、阳性药物组、蓖麻油模型组、蓖麻油中药预防组、番泻叶模型组、番泻叶中药预防组和番泻叶中药治疗组,按试验设计进行灌胃处理后,观察各组小鼠腹泻情况,针对不同药物腹泻模型采集十二指肠、盲肠观察肠道病理变化,最后采集各组盲肠内容物进行16S rDNA测序。【结果】腹泻指数显示,灌胃1 h番泻叶中药预防组和中药治疗组均极显著(P<0.01)低于番泻叶模型组;蓖麻油模型灌胃2 h与正常对照组差异极显著。肠道病理变化主要为制造腹泻模型后黏膜下层有少量炎性细胞浸润,除正常对照组外其他组肌层和浆膜变薄。盲肠Alpha测序显示,蓖麻油造模后,中药预防组能显著提高盲肠菌群丰度和多样性;Beta多样性PCoA分析表明,蓖麻油模型组与其他组样本明显分开;物种分类显示,2种药物腹泻模型处理后中药预防组和治疗组均不同程度提高拟杆菌门(Bacteroidetes)、软壁菌门(Tenericutes)丰度,降低变形菌门(Proteobacteria)丰度;属水平在正常范围内,降低乳杆菌(Lactobacillus)和大肠杆菌(Escherichia)丰度,提高有益菌Mucispirillum和拟普雷沃菌属(Alloprevotella)丰度。【结论】蓖麻油和番泻叶小鼠腹泻模型使用中药复方石苦秦散预防和治疗后,可明显改善肠道微生物丰度,特别是炎症反应相关的微生物,说明石苦秦散可通过调整肠道炎症修复发挥止泻作用。

Design of 16S rRNA gene primers for 454 pyrosequencing of the human foregut microbiome

AIM:To design and validate broad-range 16S rRNA primers for use in high throughput sequencing to classify bacteria isolated from the human foregut microbiome.METHODS:A foregut microbiome dataset was constructed using 16S rRNA gene sequences obtained from oral,esophageal,and gastric microbiomes produced by Sanger sequencing in previous studies represented by 219 bacterial species.Candidate primers evaluated were from the European rRNA database.To assess the effect of sequence length on accuracy of classification,16S rRNA genes of various lengths were created by trimming the full length sequences.Sequences spanning various hypervariable regions were selected to simulate the amplicons that would be obtained using possible primer pairs.The sequences were compared with full length 16S rRNA genes for accuracy in taxonomic classification using online software at the Ribosomal Database Project (RDP).The universality of the primer set was evaluated using the RDP 16S rRNA database which is comprised of 433 306 16S rRNA genes,represented by 36 phyla.RESULTS:Truncation to 100 nucleotides(nt)downstream from the position corresponding to base 28 in the Escherichia coli 16S rRNA gene caused misclassification of 87(39.7%)of the 219 sequences,compared with misclassification of only 29(13.2%)sequences with truncation to 350 nt.Among 350-nt sequence reads within various regions of the 16S rRNA gene,the reverse read of an amplicon generated using the 343F/798R primers had the least(8.2%)effect on classification.In comparison,truncation to 900 nt mimicking single pass Sanger reads misclassified 5.0%of the 219 sequences.The 343F/798R amplicon accurately assigned 91.8%of the 219 sequences at the species level.Weighted by abundance of the species in the esophageal dataset,the 343F/798R amplicon yielded similar classification accuracy without a significant loss in species coverage(92%).Modification of the 343F/798R primers to 347F/803R increased their universality among foregut species.Assuming that a typicalpolymerase chain reaction can tolerate 2 mismatches between a primer and a template,the modified 347F and 803R primers should be able to anneal 98%and 99.6%of all 16S rRNA genes in the RDP database.CONCLUSION:347F/803R is the most suitable pair of primers for classification of foregut 16S rRNA genes but also possess universality suitable for analyses of other complex microbiomes.

High-throughput sequencing of 16S r RNA amplicons characterizes gut microbiota shift of juvenile sea cucumber Apostichopus japonicus feeding with three antibiotics

Sea cucumber Apostichopus japonicus is an important marine economic species in Asian countries due to its profound nutritional and medicinal value. So far, with the rapid development of intensifi ed artifi cial aquaculture of sea cucumbers, the use of antibiotics is still an inexpensive and dispensable way to treat pathogenic infections, especially during the nursery phase. However, there is little information on the eff ects of antibiotics on the intestinal microbiota of sea cucumber. Therefore an Illumina based sequencing method was used to examine the intestinal bacterial composition of juvenile A . japonicas following diets with three typical antibiotics (tetracycline, erythromycin, and norfl oxacin) under 15, 30, and 45 d. The fi ndings reveal that diff erent antibiotics have distinct eff ects on the growth performance of juvenile sea cucumbers. However, the richness and diversity of microbiota were barely aff ected by antibiotics but the community composition alterations indicated that the three antibiotics exhibited their respective patterns of reshaping the intestinal bacteria of juvenile sea cucumbers. In common, the abundance of some sensitive genera with helpful functions, such as Thalassotalea , Shewanella , Sulfi tobacter , and Halomonas decreased signifi cantly with exposure to antibiotics and the abundance of multiple potential pathogenic- and suspected antibiotic-resistant microorganisms like Arcobacter , Leucothrix , and Clostridium_sensu_stricto_1 was found increased signifi cantly in the antibiotic groups. These results suggest that low doses of antibiotics could aff ect the composition of the intestinal microbiota of sea cucumbers and might increase the risk of infection of the hosts. This study could help us to explore how antibacterial compounds modify the gut microbiota of sea cucumbers and provide theoretical guidance in hatchery management by scientifi c antibiotic use in sea cucumber mariculture.

基于16S rRNA测序分析阻塞性睡眠呼吸暂停患者肠道靶标菌群的变化

目的分析阻塞性睡眠呼吸暂停(OSA)患者和健康人群肠道菌群的差异,探讨肠道菌群在OSA发病中的作用及意义。方法随机纳入2022年1月~12月就诊于本院诊断为OSA的患者39例作为OSA组,健康志愿者20例作为对照组。收集两组人群的粪便标本,通过16SrRNA高通量测序分析其微生物组成,分析两组人群肠道菌群之间的Alpha多样性、Beta多样性、物种差异与标志物种和差异生物功能代谢通路功能预测分析。结果Alpha多样性分析显示,OSA组的物种多样性指数Shannon和Simpson、物种丰度指数Observedspecies及菌群均匀度指数Pielou均低于对照组(P<0.05);Beta多样性分析显示,两组间群落结构存在差异(P<0.05)。OSA组肠道菌群群落结构上与对照组存在差异,潜在致病菌属如假单胞菌属、巨单胞菌属等菌群丰度增加(P<0.05)。LEfSe分析结果显示,与对照组相比,OSA组假单胞菌属、巨单胞菌属、梭杆菌属丰度升高(P<0.05)。关联网络分析结果显示,影响宿主稳态的为差异标志菌群;随机森林分析和ROC曲线结果显示,假单胞菌属是具有重要鉴别意义的生物标志菌属。差异代谢通路预测功能显示,维持肠道菌群稳态起主要作用功能的是生物合成功能,假单胞菌属参与了芳香族生物胺降解和酮葡萄糖酸代谢。结论OSA患者存在肠道菌群紊乱,肠菌中假单胞菌属可能通过参与物质代谢影响OSA发生发展,可望作为防治OSA的潜在肠菌靶标。

Fecal microbiota of three bactrian camels(Camelus ferus and Camelus bactrianus) in China by high throughput sequencing of the V3-V4 region of the 16S rRNA gene

This study aimed to reveal the microbial diversity in the fecal samples of bactrian camels using the 16 S r RNA sequencing analysis on the Illumina Mi Seq platform. Three fecal samples were collected from two geographical regions in China. Operational taxonomic unit（OTU） clustering was performed by identifying an OTU at 97% sequence identity. The alpha and beta diversities were applied to estimate the differences in microbial diversity among the three fecal samples. Totally, 4409, 3151 and 4075 OTUs in the fecal samples were identified in the Lop Nor wild camel（Camelus ferus）, the domestic camel（C. bactrianus） and Dunhuang wild camel（C. ferus）, respectively. The majority of bactreria were affiliated with phylum Firmicutes and Bacteroidetes in the three samples. The wild camels had higher gastrointestinal tract microbial diversity than the domestic one, while the microbial composition of the Lop Nor wild camel shared higher similarity with domestic camel at the genus and family levels than that of the Dunhuang wild camel did. Our results may provide a theoretical basis for assessing their health conditions and may thus be useful for protecting the critically endangered species of C. ferus.

基于16S rDNA测序分析不同剩余采食量皖南黄兔盲肠中微生物菌落多样性

[目的]本研究旨在探究皖南黄兔个体间剩余采食量水平差异与盲肠菌群多样性的关系。[方法]选取体重、年龄相近的110只皖南黄兔仔兔进行60 d的饲养试验,试验结束后测定个体剩余采食量(residual feed intake, RFI),根据RFI值选出5只高RFI和5只低RFI皖南黄兔母兔,分别为H组(高RFI值)和L组(低RFI值)。利用16S rDNA扩增子测序技术对H组和L组母兔盲肠内容物进行比较分析。[结果]不同剩余采食量水平的家兔盲肠菌群组成丰富度无显著差异(P>0.05),而菌落多样性存在一定差异。L组黄兔盲肠微生物中纺锤链杆属(Fusicatenibacter)、经黏液真杆菌属(Blautia)、毛螺菌属(Lachnospira)、草酸杆菌属(Oxalobacter)、肠杆菌属(Intestinibacter)的相对丰度显著高于H组(P<0.05);Paludicola、Mailhella、螺杆菌属(Helicobacter)、假单胞菌属(Pseudomonas)的相对丰度显著低于H组(P<0.05)。[结论]不同RFI水平的皖南黄兔群体盲肠菌群多样性和结构组成存在一定差异,包括毛螺菌属、纺锤链杆菌属、结肠杆菌等与血脂代谢、血糖调节过程相关的菌群在两组间的丰度存在显著差异。调节肠道菌群的结构及丰度可能是改善家兔饲料利用效率的有效途径。

基于16S rRNA基因测序技术分析急性肝内胆汁淤积大鼠的肠道菌群

目的 探讨ANIT诱导的大鼠急性肝内胆汁淤积(acute intrahepatic cholestasis,AIC)与肠道菌群的关系。方法 以SPF级的SD大鼠为实验对象,随机分为正常对照组、模型组,每组24只,依据取材时间不同又将各组随机分为24 h、48 h、72 h 3个亚组,每组8只。模型组通过一次性予2%ANIT按100 mg·kg^(-1)·d^(-1)灌胃造模诱发AIC模型,正常对照组予等容积色拉油灌胃。通过高通量测序法对肠道粪便细菌16S rRNA的V_(3)~V_(4)区进行基因测序及生物信息学分析。结果 AIC造模后,随着时间增加,α多样性指数逐步升高,造模72 h组相比造模24 h组,Shannon指数有显著性差异;β多样性发现,利用基于加权Unifrac距离PcoA分析或NMDS分析均显示:正常对照组、造模24 h组和造模48 h组的肠道菌群显示比较明显的聚集效应;在门水平上,厚壁菌门(Firmicutes)丰度在造模48 h后显著下降,变形菌门(Proteobacteria)的丰度显著上升,拟杆菌门(Bacteroidetes)的丰度显著上升;LEfSe对组间差异显著的物种分析结果显示,LDA值大于4的微生物类群有35个。结论 模型组干预后大鼠肠道菌群结构和多样性均发生显著变化。

基于UPLC-Q-Orbitrap HRMS代谢组学和16S rRNA基因测序探讨骨疏丹补肾机制

目的 整合代谢组学和肠道微生物组学的研究策略探讨骨疏丹(Gushudan, GSD)预防氢化可的松诱导的肾阳虚证(kidney-yang deficiency syndrome, KYDS)大鼠的补肾作用机制。方法 分别采用超高效液相色谱-四级杆/静电场轨道阱高分辨质谱(UPLC-Q-Orbitrap HRMS)的非靶向代谢组学和16S rRNA基因测序分析的肠道微生物组学方法,分析正常对照组、肾阳虚证模型组、骨疏丹给药组和阳性对照组大鼠粪便代谢物谱与肠道菌群组成,采用Pearson相关分析探讨内源性差异代谢物与差异菌群之间的相关性。结果 基于UPLC-Q-Orbitrap HRMS的代谢组学方法在正负离子模式下共发现骨疏丹参与调控肾阳虚症的22种差异代谢物,如色氨酸、鹅去氧胆酸、肌酐和油酸酰胺等,主要涉及氨基酸代谢、胆汁酸代谢、能量代谢和脂质代谢。基于16S rRNA测序分析发现骨疏丹在属水平显著上调普雷沃氏菌(Prevotellaceae)的相对丰度(P<0.05),显著下调颤杆菌(Oscillibacter)的相对丰度(P<0.05)。相关性分析结果表明甘胆酸和鹅去氧胆酸与在属水平显著改变的普雷沃氏菌(Prevotellaceae)显著正相关(P<0.05),而与考拉杆菌(Phascolarctobacterium)显著负相关(P<0.05)。二十二碳六烯酸与毛螺菌(Lachnospiraceae)显著负相关(P<0.05)。结论 骨疏丹通过良性调节内源性代谢和肠道菌群结构发挥补肾作用,为中药通过肠-肾轴治疗疾病提供新的思路和方法。

基于16S rRNA测序的1型和2型糖尿病大鼠肠道菌群差异

通过16S rRNA测序分析T1DM与T2DM大鼠肠道菌群的差异。Alpha多样性分析显示,T1DM组和T2DM组肠道菌群多样性和均匀度均高于对照组,Beta多样性分析显示不同组样本间存在差异且有很好的聚类。在门水平上,T1DM和T2DM大鼠肠道菌群以Firmicutes和Bacteroidetes为主,Firmicutes/Bacteroidetes(F/B)比例相比于对照组增加,且T2DM组高于T1DM组。与对照组相比,T1DM组Actinobacteria丰度显著增加,Anaerofustis丰度显著降低。在T2DM大鼠中,Proteobacteria、Oscillospira丰度显著增加。Oscillospira丰度在T2DM组增加,在T1DM组无明显差异。Rhodococcus为T1DM组的特有属,Coprobacillus和Roseburia为T2DM组的特有属。研究结果表明大鼠肠道菌群失调与T1DM和T2DM密切相关,可为T1DM和T2DM患者的个性化治疗提供科学依据。

16S-rRNA测序技术分析早产儿肠道细菌基因组指导新生儿坏死性小肠结肠炎手术时机选择的研究

目的探讨16S-rRNA测序技术分析早产儿肠道细菌基因组指导新生儿坏死性小肠结肠炎(NEC)手术时机选择。方法前瞻性选择2021年1月至2022年6月百色市人民医院需要手术治疗的NEC患儿30例为观察组,选择同期内科保守治疗的Ⅰ期15例和Ⅱa期15例患者为对照组。采用HiSeq测序平台,借助双端测序模式进行高通量二代测序,比较两组多样性指数、优势均属丰度及不同优势菌比值等;绘制受试者操作特征(ROC)曲线,分析16S-rRNA测序技术的指导价值。结果60例患者60份样本中共获得细菌84个,且两组样品均为副杆状菌属最高,其次为Ruminococcus、Blautia、Aeromonas和Fusobacterium;两组肠道菌群上述菌门丰度存在差异(P<0.05);从粪便标本中共获得有效序列7347481条,人均130857条,测序平均覆盖度为(92.15±5.61)%;观察组手术治疗的NEC患儿中香农-维纳(Shannon)及辛普森多样性(Simpson)指数低于对照组内科保守治疗患儿,差异有统计学意义(P<0.05);ROC曲线结果表明,16S-rRNA测序技术在NEC患儿手术时机选择中的指导AUC为0.846,指导灵敏度为87.51%,特异度为83.16%。结论NEC患儿常伴有肠道细菌基因组改变,且菌群结构的变化与患儿病情严重程度有关,通过16S-rRNA测序技术能指导NEC患儿手术治疗时机。

Evaluation of Real-Time 16S rDNA PCR and Pyrosequencing for Routine Identification of Bacteria in Joint Fluid and Tissue Specimens

16S rDNA PCR and sequencing are powerful tools for bacterial detection and identification, although their routine use is not currently widespread in the field of clinical microbiology. The availability of pyrosequencing now makes 16S rDNA assays more accessible to routine diagnostic laboratories, but this approach has had limited evaluation in general diagnostic practice. In this study we evaluated a real-time 16S rDNA PCR and pyrosequencing assay for use in a routine microbiology laboratory, by retrospectively testing joint fluid and joint tissue specimens received for conventional culture. We found that use of the real-time 16S rDNA assay was clinically valuable in this specimen type because it enabled us to identify a small number of culture-negative infections. Although faster and less labour-intensive, we found that the utility of pyrosequencing for pathogen identification is still hampered by shorter read lengths compared to conventional (Sanger) sequencing. Combining results from both molecular and conventional culture methods, bacteria were only detected in 11.8% specimens in this study. However, the detection rate was increased to 18.6% if specimens were only included from patients with a documented clinical suspicion of infection. In conclusion, while pyrosequencing had significant advantages in speed and ease-of-use over conventional sequencing, multiple reactions will be required to deliver comparable species-level identification, thus negating many of the benefits of using the technique. We found that 16S rDNA PCR and sequencing should be rationally targeted on the basis of good clinical information in the routine diagnostic setting, and not used as a general screening test for the exclusion of bacterial infection in joint specimens.

Application of 16S rDNA Sequencing Technology in the Analysis of Pathogenic Bacteria in Sputum of Severe Pneumonia

The diagnosis of pathogenic bacteria in severe pneumonia is difficult and the prognosis is poor. Its outcome is closely related to bacterial pathogenicity and the timeliness and pertinence of antibiotic treatment. Therefore, early diagnosis is of great significance to the prognosis of patients. Sputum examination and culture is the gold standard for the diagnosis of pathogens of severe pneumonia. However, due to the long time of bacterial culture, the early use of antibiotics, the change of bacteria species, mixed infection and other problems, the results of bacterial culture in sputum are often false negative. With the continuous application of new molecular biology techniques in clinical detection, the classification of bacteria and microorganisms has deepened from the identification of phenotypic characteristics to the classification of gene characteristics. Sequencing analysis with 16S rDNA sequencing technology has the characteristics of high sequencing flux, large amount of data obtained, short cycle, and can more comprehensively reflect the species composition of microbial community, real species distribution and abundance information. In this paper, 16S rDNA sequencing technology was used to analyze the bacterial population composition in the sputum of severe pneumonia, and to explore a new method of etiological diagnosis.

Application of 16s rDNA Sequencing in the Analysis of Pathogenic Bacteria in Sputum of Severe Bacterial Pneumonia

Objective: 120 patients with severe pneumonia who were kept in the comprehensive ICU of our hospital in 2018 were selected, and 16s rDNA sequencing was performed to analyze the composition of pathogenic bacteria in the sputum of severe pneumonia. Methods: The sputum samples of patients with severe bacterial pneumonia were collected, and the diversity of pathogens in the samples was analyzed by polymerase chain reaction (PCR) amplification and high-throughput sequencing (16s rDNA PCR-DGGE). Results: Sequence showed that sputum samples contained a relatively large number of species, and there were many species that were not detected by sequencing. The dominant bacteria were Streptococcus, Sphingomonas, Corynebacterium, Denatobacteria, Aquobacteria, Acinetobacteria, Prevotella, Klebsiella, Pseudomonas, etc. Conclusion: Bacteria caused by sputum of severe bacterial pneumonia are complex and diverse, which provides new methods and ideas for individualized treatment of patients with severe pneumonia.

基于16S rRNA基因测序分析烟叶发酵过程中表面微生物的多样性

雪茄烟叶经过发酵处理才能制成风味独特的雪茄产品,研究烟叶发酵过程中微生物的多样性对提高国产雪茄烟叶品质有重要意义。作者采集了来自云南省8个地区初步晾晒的雪茄烟叶和7种来自多米尼加共和国各地的优质雪茄烟叶,基于16S rRNA基因测序技术鉴定各个样品表面微生物种类,通过OTU聚类分析、Alpha多样性分析、物种组成分析等方法进行微生物多样性分析,获得晾晒烟叶表面的优势菌。结果显示,云南省雪茄烟叶的优势菌属为葡萄球菌属,其他常见菌属有假单胞菌属、无色杆菌属、棒状杆菌属、泛菌属等;多米尼加共和国雪茄样品中微生物物种分布较中国雪茄样品更均匀,微生物的多样性整体高于中国雪茄样品,其优势菌属多为葡萄球菌属、棒状杆菌属、四联球菌属;云南省临沧市雪茄烟叶优势菌属与多米尼加雪茄相似,而且它的物种多样性和均匀度高于中国其他地区,具有较大的发展潜力。

基于16S rDNA测序分析云南高原世居汉族高血压患者肠道菌群的差异

目的:采用16S rDNA测序技术分析云南高原世居汉族新诊断高血压患者与健康人群肠道菌群的差异。方法:基于中国多民族队列(CMEC)的基线调查数据,将云南丽江永胜县汉族新诊断高血压患者分为未用药组(H组)23例,高血压药物治疗组(HM组)13例,以健康人群26例作为对照(C组)。收集3组血液、粪便样本,用于血液生化指标检测及16S r DNA测序技术分析。结果:α多样性结果显示,与C组相比,H组肠道菌群物种丰富度下降(P=0.037);β多样性结果显示,3组比较差异具有统计学意义(R^(2)=0.047,P=0.043),其中HM组与C组差异最为显著(R^(2)=0.0552,P=0.005)。在属水平上,与C组比较,H组双歧杆菌、柯林斯杆菌属、苏黎世杆菌属等7个菌属丰度显著降低,而肠球菌属丰度显著增高(均P<0.05)。H组与C组Spearman相关性分析结果显示,双歧杆菌属和柯林斯杆菌属与收缩压(SBP)和舒张压(DBP)呈负相关关系(P<0.05)。受试者工作特征(ROC)曲线分析显示,双歧杆菌和柯林斯杆菌作为高血压疾病的微生物标志物的曲线下面积(AUC)为0.78。结论:云南高原世居汉族人群高血压疾病的发生、发展与肠道菌群紊乱相关,表现为有益菌减少,有害菌增加;双歧杆菌和柯林斯杆菌或可作为高原地区人群高血压的微生物标志物,补充有益菌和调整饮食结构可预防和改善高血压。

基于16S rDNA测序分析柔嫩艾美耳球虫感染对鸡盲肠菌群的影响

为探究柔嫩艾美耳球虫感染对鸡盲肠菌群的影响,采集对照组和球虫感染组雏鸡盲肠内容物,提取DNA,16S rDNA测序分析鸡盲肠菌群丰度、多样性的变化,采用粪菌移植验证正常鸡肠道菌群对柔嫩艾美耳球虫感染的保护作用。16S rDNA测序结果显示,柔嫩艾美耳球虫感染后盲肠内菌群丰度、多样性显著降低。在门水平上,感染前,两组雏鸡盲肠的优势菌门均为厚壁菌、变形菌、放线菌;感染后厚壁菌门丰度下降,变形菌门、放线菌门的丰度上升,差异显著(P<0.05),感染组出现了拟杆菌门。在属水平上,与对照组相比,感染组鸡盲肠微生物菌群中乳杆菌属、埃希菌-志贺菌属、拟杆菌属、罗姆布茨菌属、棒状杆菌属、肠球菌属、变形杆菌属丰度增加但差异不显著(P>0.05),毛螺菌科未定属、瘤胃球菌属、梭菌UCG-014、瘤胃球菌科未定属丰度显著下降(P<0.05),GCA-900066575丰度降低但差异不显著(P>0.05)。粪菌移植结果显示,与生理盐水移植组相比,盲肠内容物移植组和粪便菌移植组能显著减轻柔嫩艾美耳球虫感染后鸡增重降低的影响,减轻盲肠损伤,卵囊产量显著下降(P<0.05)。盲肠内容物移植组和粪便菌移植组的抗球虫指数分别为173.12和164.38,可以抵抗柔嫩艾美耳球虫的感染。以上结果表明,柔嫩艾美耳球虫感染破坏了原始肠道菌群,鸡盲肠的组成和结构被改变,促进了以埃希菌-志贺菌为代表的致病菌快速生长和定殖,而正常鸡肠道内存在益生菌,可保护机体对抗柔嫩艾美耳球虫感染。

Correlation analysis of breast fibroadenoma and the intestinal flora based on 16S rRNA sequencing

Objective To analyze the characteristics of the intestinal microflora in patients with breast fibroadenoma using 16S ribosomal RNA(rRNA)high-throughput sequencing.Methods Fecal samples from 20 patients with breast fibroadenoma and 36 healthy subjects were randomly collected and analyzed using high-throughput sequencing technology for 16S rRNA V4 region sequencing,and the alpha diversity(Chao index,Shannon index)was calculated using Mothur(v.1.39.5)software.Beta diversity was analyzed using QIIME(v1.80).SPSS software(version 23.0)and the t-test of two independent samples were used to analyze differences in the abundance of bacteria between the two groups.Results Compared with that in the healthy control group,theαdiversity of the intestinal microflora in breast fibroadenoma patients increased,but the difference was not statistically significant(P>0.05).At the phylum level,significant differences were observed between the two groups.The abundance of Firmicutes was higher in the breast fibroadenoma group(P<0.05),whereas the abundance of Synergistetes was higher in the healthy control group(P<0.005).A total of five bacterial genera showed significant differences between the two groups:the breast fibroadenoma group showed higher levels of Bautia(P<0.005),Coprococcus(P<0.005),Roseburia(P<0.05),and Ruminococcus(P<0.005),whereas Sutterella was more abundant in the healthy control group than in the breast fibroadenoma group(P<0.05).Conclusion The diversity and abundance of the intestinal flora in patients with breast fibroadenoma are significantly different from those in healthy subjects,suggesting that an imbalance in the intestinal flora is correlated with the occurrence of breast fibroadenoma.

基于全长16S rRNA测序的电磁辐射环境人群口腔菌群特征研究

目的:描述和比较工作环境中存在电磁辐射人群与普通工作环境人群口腔菌群分布特征及差异,为电磁辐射损伤的早期防护提供潜在筛查方法及依据。方法:2023年10~12月期间随机抽取某两家单位工作人员共130人为研究对象,依据其工作环境分为电磁辐射工作环境组(JMS组,n=65)与普通工作环境组(BJ组,n=65),收集两组口腔唾液标本并提取DNA,通过高通量基因测序平台对其16S rRNA进行测序,研究并比较两组口腔菌群构成特征。结果:β多样性分析结果表明JMS组与BJ组口腔菌群结构存在统计学差异(P<0.01)。口腔菌群构成中,在门水平上,两组拟杆菌门、梭杆菌门的相对丰度存在统计学差异(P<0.05);在纲水平上,两组芽孢杆菌纲、梭杆菌纲、梭菌纲的相对丰度均有极显著差异(P<0.001),而放线菌纲、红蝽菌纲、蒂西耶氏菌纲和拟杆菌纲的相对丰度有显著差异(P<0.05)。LEfSe差异分析结果表明,JMS组具有独特的菌群结构,有11种菌显著富集,分别为梭菌纲、微黄奈瑟氏球菌A、管道杆菌属、诺曼管道杆菌、细小口腔杆菌、梭杆菌门、梭杆菌纲、梭杆菌目、不解糖口腔杆菌、龈沟产线菌、梭杆菌科。此外,菌群功能预测结果显示,JMS组口腔菌群在免疫系统途径、抗肿瘤药耐药性途径、能量代谢途径上的富集程度高于BJ组;BJ组在碳水化合物代谢途径、膜传输途径的富集程度高于JMS组。结论:普通工作环境和电磁辐射工作环境下人群具有不同的口腔微生物特征与菌群代谢特征,且表现在多个分类水平上,提示电磁辐射工作环境可能对人口腔菌群分布特征产生影响,未来仍需进一步证实和探索。

基于16S rDNA测序技术分析实验用猫的肠道菌群特征

目的通过16S rDNA高通量测序研究不同品种和不同性别实验用猫肠道菌群的变化特征。方法选取实验用猫21只,根据实验用猫的品种和性别进行分组,按品种分组(普通组=12只、品种组=9只),按性别分组(雄性组=9只、雌性组=12只)。采取猫的粪便样本,提取DNA,PCR扩增,利用通过16S rDNA高通量测序进行菌群结构及多样性的差异分析。结果实验用猫门水平的优势菌为厚壁菌门、拟杆菌门、放线菌门(相对丰度之和大于96%)。属水平相对丰度水平较高的菌群为链球菌属、肠球菌属、欧鲁森氏菌属、普雷沃氏菌属、双歧杆菌属和柯林斯菌属。性别可对实验用猫肠道菌群的构成产生影响,且雄性猫和雌性猫之间存在丰度有统计学差异的菌种。相较于雌性猫,普雷沃氏菌属、柔嫩梭菌属和土孢杆菌属在雄性猫的肠道中的相对丰度增加(t检验,P<0.05)。结论利用16S rDNA高通量测序分析了实验用猫的肠道菌群的组成、丰度等基本特点,并阐述了性别及品种对实验用猫肠道菌群物种组成的影响。