Table 4_Multi-omics insights into the response of the gut microbiota and metabolites to albendazole deworming in captive Rhinopithecus brelichi.docx

BackgroundParasite infection and deworming treatment affect the host gut microbiota. Exploring the response mechanism of the gut microbiota in Rhinopithecus brelichi (R. brelichi) to albendazole deworming treatment is of great value for protecting this critically endangered species. Methods and resultsThis study used metataxonomics and metabolomics to explore the responses of the gut microbiota and metabolites of R. brelichi to albendazole deworming treatment. The results showed that deworming significantly reduced the eggs per gram of feces (EPG). The 16S rRNA gene sequencing results showed that the richness and diversity of the gut microbiota in R. brelichi after deworming were significantly increased. Meanwhile, deworming treatment also changed the composition of the gut microbiota. At the genus level, the Christensenellaceae R7 group, UCG 002, UCG 005, uncultured rumen bacterium, and Rikenellaceae RC9 gut group were significantly enriched in the pre-deworming samples. Unclassified Muribaculaceae, Prevotella 9, and Bacteroides were significantly enriched in the post-deworming samples. Metabolomics analysis revealed that the relative abundance of 382 out of 1,865 metabolites showed significant differences between the pre- and post-deworming samples. Among them, 103 metabolites were annotated based on the HMDB and mainly classified into Prenol lipids, Carboxylic acids and derivatives, and Organooxygen compounds, etc. The KEGG enrichment analysis result indicated that these metabolites were mainly involved in energy, amino acid, lipid, and purine metabolism. Correlation analysis showed that Bacteroides and unclassified Muribaculaceae, whose relative abundances were upregulated after deworming treatment, were positively correlated with Kaempferol, 5,7-Dihydroxy-3-methoxy-4′-prenyloxyflavone, Purpurin, and Rhein, which have anti-parasitic activities. The Christensenellaceae R7 group, with a downregulated relative abundance after deworming treatment, was not only negatively correlated with the above four metabolites, but also positively correlated with Retinyl beta-glucuronide, which is a storage form of vitamin A, and positively correlated with CDP-Choline, which increases the host’s susceptibility to Entamoeba histolytica and Plasmodium falciparum. ConclusionThis study emphasizes that deworming treatment has an impact on the gut microbiota and metabolic functions of R. brelichi. By exploiting the correlations between differential microbiota and metabolites, potential probiotics or prebiotics can be explored, thereby enhancing the efficiency of deworming and reducing its side effects.研究背景：寄生虫感染与驱虫治疗可影响宿主肠道菌群。探究黔金丝猴（Rhinopithecus brelichi，简称R. brelichi）肠道菌群对阿苯达唑驱虫治疗的响应机制，对保护这一极度濒危物种具有重要研究价值。 研究方法与结果：本研究采用宏分类组学（metataxonomics）与代谢组学技术，剖析黔金丝猴肠道菌群及代谢物对阿苯达唑驱虫治疗的响应情况。研究结果显示，驱虫治疗可显著降低每克粪便虫卵数（eggs per gram of feces, EPG）。16S rRNA基因测序结果表明，驱虫后黔金丝猴肠道菌群的丰富度与多样性均显著提升，同时肠道菌群组成亦发生显著改变。在属水平上，Christensenellaceae R7 group、UCG 002、UCG 005、未培养瘤胃细菌（uncultured rumen bacterium）以及Rikenellaceae RC9肠道菌群组在驱虫前样本中显著富集；未分类Muribaculaceae（unclassified Muribaculaceae）、Prevotella 9与拟杆菌属（Bacteroides）则在驱虫后样本中显著富集。代谢组学分析显示，1865种代谢物中有382种在驱虫前后样本中的相对丰度存在显著差异。其中103种代谢物可通过人类代谢组数据库（Human Metabolome Database, HMDB）注释，主要归类为类异戊二烯脂质（Prenol lipids）、羧酸及其衍生物、有机氧化合物（Organooxygen compounds）等类别。京都基因与基因组百科全书（KEGG）富集分析结果表明，上述差异代谢物主要参与能量代谢、氨基酸代谢、脂质代谢与嘌呤代谢通路。相关性分析显示，驱虫后相对丰度上调的拟杆菌属与未分类Muribaculaceae，与具有抗寄生虫活性的山奈酚（Kaempferol）、5,7-二羟基-3-甲氧基-4'-异戊烯氧基黄酮（5,7-Dihydroxy-3-methoxy-4′-prenyloxyflavone）、紫铆因（Purpurin）以及大黄酸（Rhein）呈显著正相关。而驱虫后相对丰度下调的Christensenellaceae R7 group，不仅与上述四种代谢物呈显著负相关，还与维生素A的储存形式视黄基β-葡萄糖醛酸苷（Retinyl beta-glucuronide）呈正相关，同时也与可增加宿主对溶组织内阿米巴与恶性疟原虫易感性的CDP-胆碱（CDP-Choline）呈正相关。 研究结论：本研究证实，驱虫治疗可对黔金丝猴的肠道菌群与代谢功能产生显著影响。通过挖掘差异菌群与差异代谢物之间的潜在关联，可筛选得到潜在的益生菌或益生元，进而提升驱虫治疗效率并降低其副作用。