摘要
目的揭示轻度无症状钩虫感染对中老年人群肠道菌群与代谢的影响,为蠕虫-肠道菌群相互作用的研究提供参考。方法以2022年10月—11月安徽省黄山市祁门县、黄山区乡村土源性线虫病监测点居民检出钩虫感染者(改良加藤厚涂片法钩虫虫卵阳性)为感染组,以钩虫虫卵阴性者为对照组。采集受检者新鲜粪样,提取肠道菌群DNA,扩增16S rRNA基因并测序。计算基于丰度的覆盖估计值(ACE)、PD-wholetree、Chao指数和Shannon指数等多样性指数,采用主坐标分析(PCoA)和不加权算术平均对方法(UPGMA)层次聚类分析的β多样性分析,比较感染组和对照组肠道菌群分布的差异,通过指示值分析筛选两组的指示性菌群。用甲醇-水萃取粪样中的代谢物,进行气相色谱-质谱(GC-MS)和液相色谱-质谱(LC-MS)双平台非靶向测序的代谢组学分析,用正交偏最小二乘方判别分析(OPLS-DA)建立筛选感染组与对照组差异代谢物的模型,制作火山图。根据P值、变量权重值和差异倍数筛选差异代谢产物。通过京都基因与基因组百科全书(KEGG)数据库检索差异代谢物的相关代谢通路,对差异代谢物进行富集分析。两个代谢物之间的线性相关程度采用Pearson相关分析。结果研究对象共22人,其中感染组11人,对照组11人。感染组平均年龄72.2岁,均为轻度无症状带虫者;对照组平均年龄53.1岁。测序共获得1755649条16S rRNA序列,经序列质控和拼接后聚类为1245条扩增序列变体。共鉴定出15个门22个纲58个目94个科191个属389个菌种。多样性指数分析结果显示,感染组肠道菌群多样性的ACE、PD-wholetree、Chao和Shannon指数分别为188.768、16.533、192.667和5.195,对照组分别为167.829、15.294、157.371和4.898,两组间差异均无统计学意义(t=1.266、0.952、1.266、0.962,均P>0.05)。PcoA分析结果显示,感染组与对照组的肠道菌群有不同程度的重叠,但仍可分为两个不同
Objective To reveal the effect of mild asymptomatic hookworm infection on intestinal microflora and metabolome in middleaged and the elderly people,to provide reference information for study on interaction of helminthintestinal microflora.Methods In October to November 2022,from the parasite infection survey in residents at the surveillance site for soiltransmitted nematodes infection in Qimen County and Huangshan District of Huangshan City,Anhui Province,the hookworm egg positives detected by modified KatoKatz thick smear method were assigned in infection group,and the hookworm egg negatives in control group.Fresh fecal samples from the examinees were collected for extraction of intestinal microflora DNA,of which 16S rRNA gene sequence was amplified by PCR and sequenced.The abundancebased coverage estimator(ACE),PDwholetree,Chao index and Shannon index were calculated.The principal coordinate analysis(PCoA)andβdiversity analysis of the unweighted pair group method with arithmetic mean(UPGMA)hierarchical clustering were used to compare the difference in the distribution of intestinal microflora between the infection group and the control group,to screen the indicator flora by indicator value analysis.The metabolites in the fecal samples were extracted with methanolwater extraction method for metabolome analysis by gas chromatographymass spectrometry(GCMS)and liquid chromatographymass spectrometry(LCMS)for nontargeted sequencing.Orthogonal partial least squares discriminant analysis(OPLSDA)was used to establish a model for screening differential metabolites between the infection group and the control group,and generate the volcano plot.The differential metabolites were screened based on Pvalues,variable weight value,and difference multiple.The Kyoto Encyclopedia of Genes and Genomes(KEGG)database was used to search the metabolic pathways of related differential metabolites.Enrichment analysis of differential metabolites was performed,and the degree of linear correlation between two metabolites was analyzed by Pea
作者
汪业彬
沈续航
邓国强
胡赛敏
汪飞
张玲玲
沈继龙
WANG Yebin;SHEN Xuhang;DENG Guoqiang;HU Saimin;WANG Fei;ZHANG Lingling;SHEN Jilong(Huangshan District Center for Disease Control and Prevention,Huangshan District 245700,Anhui,China;Department of Gastroentorology,the First Affiliated Hospital of Anhui Medical University,Hefei 230033,China;Qimen Center for Disease Control and Prevention,Qimen 245600,Anhui,China;Huangshan District Center for Disease Control and Prevention,Huangshan 245700,Anhui,China;The Key Laboratory of Zoonoses Anhui,Department of Parasitology and Microbiology,Anhui Medical University,Hefei 230032,China)
出处
《中国寄生虫学与寄生虫病杂志》
CSCD
北大核心
2024年第1期17-26,共10页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家自然科学基金(82072304)。
