元基因组分析揭示了羊茅中毒情况下牛直肠微生物组和抗菌药耐药性组的显著变化

Yihang Zhou
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摘要

牧草中毒会导致放牧受内生菌感染的高羊茅的奶牛生长和繁殖能力下降。为了描述肠道微生物群及其对羊茅中毒的反应,我们收集了八头安格斯西门塔尔怀孕母牛和小母牛在补充30天有毒羊茅种子前后的粪便样本。我们采用全基因组射枪法对 16 个元基因组进行了测序,并生成了 157 Gbp 的元基因组序列。通过重新组合和注释,我们获得了 13.1 Gbp 的参考序列,并为牛直肠微生物群鉴定了 2200 万个微生物基因。我们发现,经过有毒种子处理后,微生物多样性明显减少(P<0.01),这表明微生物组出现了菌群失调。粪便微生物群中有 6 个细菌科和 31 个物种明显增加(P-adj<0.05),其中包括瘤胃中含量最高的核心类群成员。直肠微生物群中瘤胃微生物的这种全球性增加表明,羊草中毒可能会损害瘤胃微生物群。其中,占瘤胃微生物群约 2% 的重要物种反刍球菌 P7 受到的影响最大,从 0.17% 增加到 2.8% (P<0.01),增加了 16 倍。我们假设,瘤胃反刍球菌 P7 在毒羊茅胁迫下适应了大肠环境,导致丰度急剧增加。功能富集分析表明,代表性过高的通路从能量代谢转向抗微生物和 DNA 复制。总之,我们发现在羊茅中毒中微生物群的组成、丰度和功能能力发生了剧烈变化,我们的研究结果表明反刍球菌 P7 是羊茅中毒管理的潜在生物标记物。
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Metagenomic analysis revealed significant changes in cattle rectum microbiome and antimicrobial resistome under fescue toxicosis
Fescue toxicity causes reduced growth and reproductive issues in cattle grazing endophyte-infected tall fescue. To characterize the gut microbiota and its response to fescue toxicosis, we collected fecal samples before and after a 30-days toxic fescue seeds supplementation from eight Angus Simmental pregnant cows and heifers. We sequenced the 16 metagenomes using the whole-genome shotgun approach and generated 157 Gbp of metagenomic sequences. Through de novo assembly and annotation, we obtained a 13.1 Gbp reference contig assembly and identified 22 million microbial genes for cattle rectum microbiota. We discovered a significant reduction of microbial diversity after toxic seed treatment (P<0.01), suggesting dysbiosis of the microbiome. Six bacterial families and 31 species are significantly increased in the fecal microbiota (P-adj<0.05), including members of the top abundant rumen core taxa. This global elevation of rumen microbes in the rectum microbiota suggests a potential impairment of rumen microbiota under fescue toxicosis. Among these, Ruminococcaceae bacterium P7, an important species accounting for ~2% of rumen microbiota, was the most impacted with a 16-fold increase from 0.17% to 2.8% in feces (P<0.01). We hypothesized that rumen Ruminococcaceae bacterium P7 re-adapted to the large intestine environment under toxic fescue stress, causing this dramatic increase in abundance. Functional enrichment analysis revealed that the overrepresented pathways shifted from energy metabolism to antimicrobial resistance and DNA replication. In conclusion, we discovered dramatic microbiota alterations in composition, abundance, and functional capacities under fescue toxicosis, and our results suggest Ruminococcaceae bacterium P7 as a potential biomarker for fescue toxicosis management.
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