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Induction of Subacute Ruminal Acidosis Affects the Ruminal Microbiome and Epithelium

机译:亚急性瘤胃酸中毒的诱导影响瘤胃微生物组和上皮细胞。

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Subacute ruminal acidosis (SARA) negatively impacts the dairy industry by decreasing dry matter intake, milk production, profitability, and increasing culling rate and death loss. Six ruminally cannulated, lactating Holstein cows were used in a replicated incomplete Latin square design to determine the effects of SARA induction on the ruminal microbiome and epithelium. Experimental periods were 10 days with days 1–3 for ad libitum intake of control diet, followed by 50% feed restriction on day 4, and ad libitum access on day 5 to the basal diet or the basal diet with an additional 10% of a 50:50 wheat/barley pellet. Based on subsequent ruminal pH, cows were grouped (SARA grouping; SG) as Non-SARA or SARA based on time <5.6 pH (0 and 3.4 h, respectively). Ruminal samples were collected on days 1 and 6 of each period prior to feeding and separated into liquid and solid fractions. Microbial DNA was extracted for bacterial analysis using 16S rRNA gene paired-end sequencing on the MiSeq Illumina platform and quantitative PCR (qPCR). Ruminal epithelium biopsies were taken on days 1 and 6 before feeding. Quantitative RT-PCR was used to determine gene expression in rumen epithelium. Bray–Curtis similarity indicated samples within the liquid fraction separated by day and coincided with an increased relative abundance of genera Prevotella , Ruminococcus , Streptococcus , and Lactobacillus on day 6 ( P < 0.06). Although Firmicutes was the predominant phyla in the solid fraction, a SG × day interaction ( P < 0.01) indicated a decrease on day 6 for SARA cows. In contrast, phylum Bacteroidetes increased on day 6 ( P < 0.01) for SARA cows driven by greater genera Prevotella and YRC22 ( P < 0.01). Streptococcus bovis and Succinivibrio dextrinosolvens populations tended to increase on day 6 but were not affected by SG. In ruminal epithelium, CLDN1 and CLDN4 expression increased on day 6 ( P < 0.03) 24 h after SARA induction and a tendency for a SG × day interaction ( P < 0.10) was observed for CLDN4 . Overall, results indicate more rapid adaptation to an induced bout of SARA in the solid fraction ruminal microbiome compared with ruminal epithelium.
机译:亚急性瘤胃酸中毒(SARA)通过减少干物质摄入量,牛奶产量,获利能力以及增加淘汰率和死亡人数,对乳制品业产生负面影响。在重复的不完全拉丁方形设计中使用了六只瘤胃插管的泌乳荷斯坦奶牛,以确定SARA诱导对瘤胃微生物组和上皮细胞的影响。实验期为10天,第1天至第3天为对照饮食的随意摄入,然后在第4天限制进食50%,在第5天自由摄取基础饮食或基础饮食,另外还有10%的基础饮食50:50小麦/大麦颗粒。根据随后的瘤胃pH值,根据时间<5.6 pH(分别为0和3.4 h),将母牛分组(SARA分组; SG)为Non-SARA或SARA。在进食前的每个时期的第1天和第6天收集瘤胃样品,并将其分离为液体和固体部分。使用MiSeq Illumina平台上的16S rRNA基因配对末端测序和定量PCR(qPCR)提取微生物DNA进行细菌分析。进食前第1天和第6天进行瘤胃上皮活检。定量RT-PCR用于确定瘤胃上皮中的基因表达。 Bray-Curtis的相似性表明,液体部分中的样品按天分开,并且在第6天与Prevotella,Ruminococcus,Streptococcus和Lactobacillus属的相对丰度增加吻合(P <0.06)。尽管Firmicutes是固态部分的主要门,但SG×天交互作用(P <0.01)表明SARA奶牛的第6天减少。相比之下,在较大的普雷沃特菌属和YRC22的驱动下,SARA奶牛的拟杆菌门在第6天增加(P <0.01)。牛链球菌和琥珀酸弧菌的数量在第6天趋于增加,但不受SG的影响。在瘤胃上皮中,SARA诱导后第24天,CLDN1和CLDN4表达增加(P <0.03),并且CLDN4出现SG×天相互作用的趋势(P <0.10)。总体而言,结果表明,与瘤胃上皮相比,固相瘤胃微生物组中的SARA诱导回旋更快。

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