Does the upper respiratory tract microbiome change in dairy calves fed milk replacer containing probiotics?

C. Chitko-McKown, T. McDaneld, J. Kritchevsky, K. A. Bryan, S. Eicher
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Abstract

Our overall hypothesis is that feeding probiotics to dairy calves will alter overall immunity and may additionally change the microbiome in the respiratory tract as well as the gut. A group of 20 dairy calves were split into two treatment groups: Control (N=10, milk replacer), and Probiotic (N=10, milk replacer + 0.5 g/day Bovamine Dairy). On day 0 birth weight was obtained and the calves were provided colostrum as per the dairy SOP. On day 2, probiotics were added to the milk replacer of the treated group. Blood was collected along with nasal and tonsil swabs on days 0, 7, 14, 21, 28, 42, and 48. Calves were monitored daily for fecal, nasal, and ocular discharge scores. Our preliminary data showed no significant differences (P<0.5) between the Control and Probiotic groups in immune cell populations in peripheral blood. DNA was extracted from nasal and tonsil swabs to evaluate the bacterial populations in the upper respiratory microbiome. Hypervariable regions 1 through 3 along the 16S ribosomal RNA gene were amplified by PCR and sequenced by Illumina MiSeq to determine the bacterial taxa present. Evaluation of these samples revealed a greater percentage of operational taxonomy units (OTU) were classified at the genus level in the tonsil compared to the nasal samples. The microbiome of the tonsils was more stable over time points compared to the nasal samples. Evaluation of each timepoint for nasal and tonsil samples identified bacterial taxa that changed in relative abundance with the addition of the probiotic compared to the control diet. Addition of probiotic changed the relative abundance of OTU (P<0.01) in the nasal and tonsil sampling sites and at multiple timepoints. Analyses are ongoing. Project supported by fund from the USDA Agricultural Research Service.
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饲喂含有益生菌的代乳剂后,犊牛上呼吸道微生物群是否发生变化?
我们的总体假设是,给奶牛喂食益生菌会改变整体免疫力,并可能额外改变呼吸道和肠道中的微生物群。将20头犊牛分为对照组(N=10,使用代乳剂)和益生菌组(N=10,使用代乳剂+ 0.5 g/d的牛胺乳)。在第0天获得初生体重,并按照乳制品SOP提供初乳。第2天,处理组在代乳中添加益生菌。在第0、7、14、21、28、42和48天采集血液和鼻腔和扁桃体拭子。每天监测犊牛的粪便、鼻分泌物和眼分泌物评分。我们的初步数据显示,对照组和益生菌组之间外周血免疫细胞群无显著差异(P<0.5)。从鼻腔和扁桃体拭子中提取DNA以评估上呼吸道微生物群中的细菌种群。通过PCR扩增16S核糖体RNA基因的高变区1 ~ 3,并用Illumina MiSeq测序确定存在的细菌分类群。这些样本的评估显示,与鼻样本相比,扁桃体中更大比例的操作分类单位(OTU)被分类在属水平。与鼻腔样本相比,扁桃体的微生物组随着时间点的变化更加稳定。对鼻腔和扁桃体样本的每个时间点进行评估,发现与对照组相比,添加益生菌后细菌分类群的相对丰度发生了变化。添加益生菌改变了鼻、扁桃体取样部位和多个时间点OTU的相对丰度(P<0.01)。分析正在进行中。该项目由美国农业部农业研究局资助。
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