Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.
Department of Veterinary Medicine, University of Milano, 26900, Lodi, Italy.
J Dairy Sci. 2020 Oct;103(10):9384-9406. doi: 10.3168/jds.2020-18619. Epub 2020 Aug 20.
In-depth analysis of colostrum components has identified hundreds of proteins, but data are sparse regarding their systemic uptake in the newborn calf. Moreover, heat treatment may influence these colostral components and their absorption. Our objectives were to describe the serum proteome of newborn calves before and after colostrum feeding and the possible effects of colostral heat treatment. Newborn Holstein heifer calves (n = 22) were randomized within pair and fed heat-treated (n = 11; 60°C, 60 min) or raw (n = 11) colostrum at 8.5% of birth body weight by esophageal feeder within 1 h of birth. After the single colostrum feeding, calves were not fed until after the 8-h time point, when milk was offered free-choice. Blood samples were taken immediately before feeding (0 h), as well as 4, 8, and 24 h after feeding. Whole blood packed cell volume (%), serum Brix percentage, and plasma glucose concentrations were determined for all time points. Plasma insulin and insulin-like growth factor-I concentrations were determined by radioimmunoassay for selected time points. Serum IgA and IgG were measured by radial immunodiffusion at 24 h. The serum proteome was analyzed using nano-scale reverse-phase chromatography and tandem mass spectrometry (nano LC-MS/MS) in 0- and 8-h samples. For proteomics analysis, ratios of results for 8-h to 0-h samples were analyzed with false discovery rate adjustment. For all other outcomes, repeated-measures ANOVA was performed with the fixed effects of group, time, and their interaction, and random effect of pair. Serum Brix percentage and glucose concentrations increased over time and were independent of colostrum treatment. Serum IgG and IgA concentrations at 24 h did not differ between groups. Nano LC-MS/MS identified a total of 663 unique proteins in serum, of which 261 increased in abundance, whereas 67 decreased in abundance after feeding in both groups. Among serum proteins that increased in abundance and that were previously identified in colostrum, many belonged to those involved in immune response, coagulation, the classical complement pathway, or the antimicrobial peptide class of cathelicidins. Serum proteins that decreased in abundance and that were identified in colostrum belonged to the alternative complement pathway and the membrane attack complex. Thirty-eight proteins differed in calves that were fed heat-treated colostrum compared with those fed raw colostrum. Decreased abundances in calves fed heat-treated colostrum included several enzymes involved in glycolysis or glycogenolysis, whereas the incretin gastric inhibitory polypeptide and serum insulin were increased in this group. Our findings point to important innate immune defense pathways associated with colostrum ingestion in newborn calves. Furthermore, calves fed heat-treated colostrum showed differences in serum proteins and enzymes associated with carbohydrate metabolism.
对初乳成分进行深入分析已经确定了数百种蛋白质,但有关这些蛋白质在新生牛犊体内系统性吸收的数据却很少。此外,热处理可能会影响这些初乳成分及其吸收。我们的目的是描述新生荷斯坦小母牛在初乳喂养前后的血清蛋白质组,并研究初乳热处理的可能影响。将 22 头新生荷斯坦小母牛按配对随机分组,出生后 1 小时内通过食管饲喂器以 8.5%的出生体重分别饲喂热处理(60°C,60 分钟)(n = 11)或未处理(n = 11)的初乳。在单次初乳喂养后,直到 8 小时时间点才再次给小牛喂奶,此时自由提供牛奶。在喂养前(0 小时)以及喂养后 4、8 和 24 小时采集血液样本。所有时间点均测定全血红细胞压积(%)、血清 Brix 百分比和血浆葡萄糖浓度。在选定的时间点通过放射免疫测定法测定血浆胰岛素和胰岛素样生长因子-I 浓度。在 24 小时时通过放射免疫扩散法测定血清 IgA 和 IgG。使用纳米级反相色谱和串联质谱(nano LC-MS/MS)在 0 小时和 8 小时样本中分析血清蛋白质组。对于蛋白质组学分析,使用错误发现率调整分析了 8 小时与 0 小时样本结果的比值。对于所有其他结果,使用组、时间及其相互作用的固定效应和对的随机效应进行重复测量方差分析。血清 Brix 百分比和葡萄糖浓度随时间增加,与初乳处理无关。24 小时时两组间血清 IgG 和 IgA 浓度无差异。nano LC-MS/MS 在血清中总共鉴定出 663 种独特的蛋白质,其中 261 种在两组中丰度增加,而 67 种在喂养后丰度降低。在丰度增加且在初乳中被鉴定出的血清蛋白质中,许多属于参与免疫反应、凝血、经典补体途径或抗菌肽类 cathelicidins 的蛋白质。在丰度降低且在初乳中被鉴定出的血清蛋白质属于替代补体途径和膜攻击复合物。与喂养未处理初乳的小牛相比,喂养热处理初乳的小牛中有 38 种蛋白质的丰度不同。在喂养热处理初乳的小牛中,有几种参与糖酵解或糖原分解的酶含量减少,而这组小牛中的肠降血糖素胃抑制肽和血清胰岛素含量增加。我们的研究结果表明,与新生小牛摄入初乳有关的重要先天免疫防御途径。此外,喂养热处理初乳的小牛在与碳水化合物代谢相关的血清蛋白质和酶方面表现出差异。