Characterization of the preweaned Holstein calf fecal microbiota prior to, during, and following resolution of uncomplicated gastrointestinal disease

R. Claus-Walker, G. Slanzon, L. A. Elder, H. R. Hinnant, C. M. Mandella, L. Parrish, S. C. Trombetta, C. McConnel
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Abstract

Little is known about shifts in the fecal microbiome of dairy calves preceding and following the incidence of gastrointestinal disease. The objective of this cohort study was to describe the fecal microbiome of preweaned dairy calves before, during, and after gastrointestinal disease. A total of 111 Holstein dairy calves were enrolled on 2 dairies (D1 and D2) and followed until 5 weeks old. Health assessments were performed weekly and fecal samples were collected every other week. Of the 111 calves, 12 calves from D1 and 12 calves from D2 were retrospectively defined as healthy, and 7 calves from D1 and 11 calves from D2 were defined as diarrheic. Samples from these calves were sequenced targeting the 16S rRNA gene and compared based on health status within age groups and farms: healthy (0–1 week old) vs. pre-diarrheic (0–1 week old), healthy (2–3 weeks old) vs. diarrheic (2–3 weeks old), and healthy (4–5 weeks old) vs. post-diarrheic (4–5 weeks old) calves. Healthy and diarrheic samples clustered together based on age rather than health status on both farms. Based on linear discriminant analysis, a few species were identified to be differently enriched when comparing health status within age groups and farm. Among them, Bifidobacterium sp. was differently enriched in pre-diarrheic calves at D1 (0–1 week old) whereas healthy calves of the same age group and farm showed a higher abundance of Escherichia coli. Bifidobacterium sp. was identified as a biomarker of fecal samples from healthy calves (2–3 weeks old) on D1 when compared with diarrheic calves of the same age group and farm. Feces from diarrheic calves on D2 (2–3 weeks old) were characterized by taxa from Peptostreptococcus and Anaerovibrio genera whereas fecal samples of age-matched healthy calves were characterized by Collinsella aerofaciens and Bifidobacterium longum. After resolution of uncomplicated diarrhea (4–5 weeks old), Collinsella aerofaciens was more abundant in D2 calves whereas Bacteriodes uniformis was more abundant in D1 calves. Taken together, these findings suggest that the age of the preweaned calf is the major driver of changes to fecal microbiome composition and diversity even in the face of uncomplicated gastrointestinal disease.
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断奶前荷斯坦犊牛粪便微生物区系在无并发症胃肠道疾病发生前、发生期间和缓解后的特征描述
人们对乳牛肠胃疾病发生前后粪便微生物群的变化知之甚少。这项队列研究的目的是描述断奶前乳牛在胃肠道疾病之前、期间和之后的粪便微生物组。两家牧场(D1 和 D2)共登记了 111 头荷斯坦奶牛犊牛,并对其进行随访至 5 周大。每周进行一次健康评估,每隔一周收集一次粪便样本。在 111 头犊牛中,12 头来自 D1 的犊牛和 12 头来自 D2 的犊牛被回顾性地定义为健康犊牛,7 头来自 D1 的犊牛和 11 头来自 D2 的犊牛被定义为腹泻犊牛。对这些犊牛的样本进行 16S rRNA 基因测序,并根据各年龄组和养殖场的健康状况进行比较:健康犊牛(0-1 周龄)与腹泻前犊牛(0-1 周龄)、健康犊牛(2-3 周龄)与腹泻犊牛(2-3 周龄)、健康犊牛(4-5 周龄)与腹泻后犊牛(4-5 周龄)。健康犊牛和腹泻犊牛样本根据年龄而非健康状况聚集在一起。根据线性判别分析,在比较不同年龄组和养殖场的健康状况时,确定了几个富集程度不同的物种。其中,双歧杆菌在 D1(0-1 周龄)腹泻前犊牛中的富集程度不同,而在同一年龄组和养殖场的健康犊牛中,大肠埃希氏菌的富集程度较高。与同一年龄组和养殖场的腹泻犊牛相比,D1(2-3 周龄)健康犊牛粪便样本中的双歧杆菌被鉴定为生物标志物。D2(2-3 周龄)腹泻犊牛粪便的特征是粪链球菌属(Peptostreptococcus)和厌氧弧菌属(Anaerovibrio)的类群,而年龄匹配的健康犊牛粪便样本的特征是气雾科林斯菌(Collinsella aerofaciens)和长双歧杆菌(Bifidobacterium longum)。无并发症腹泻缓解后(4-5 周龄),D2 级犊牛的铜绿菌更多,而 D1 级犊牛的均匀杆菌更多。综上所述,这些研究结果表明,断奶前犊牛的年龄是粪便微生物组组成和多样性变化的主要驱动因素,即使在无并发胃肠道疾病的情况下也是如此。
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