Technical Note: Different analytical approaches in the quantification of microbial crude protein pool in abomasal digesta of nursling goat kids

IF 2.5 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Feed Science and Technology Pub Date : 2025-05-26 DOI:10.1016/j.anifeedsci.2025.116399

Jardeson de Souza Pinheiro , Paulo Sergio Dornelas Silva , Isadora Emily Ribeiro , Dhones Rodrigues Andrade , Natalia Veloso Trópia , Yame Fabres Robaina Sancler-Silva , Erollykens Ferreira Santos , Giarlã Cunha da Silva , Simone Eliza Facione Guimarães , Marcos Inacio Marcondes

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引用次数: 0

Abstract

We evaluated methodologies to quantify microbial crude protein (MCP) pool in abomasum samples of pre-weaned dairy goat kids fed with milk replacer (MR). Sixty newborn male goat kids of the Saanen and Swiss Alpine breeds at an average age of 2 ± 1 days, and an average body weight (BW) of 3.834 ± 0.612 kg were included in the study. The goat kids received different liquid diet levels categorized as low nutritional plan (LNP; providing 1 liter of MR/goat kid/day), and a high nutritional plan (HNP; at 2 liters of MR/goat kid/day). All goat kids were harvested with 45 days of life at six-time points after feeding. Abomasum contents were sampled to determine the microbial pool and to estimate the MCP. Four methods were adopted for MCP quantification: yeast RNA – method 1, Bradford Blue – method 2 (BBM), Flow cytometry – method 3 (FCM; method proposed in the current study), and Polymerase Chain Reaction -method 4 (PCRM; method proposed in the current study). All four methods were compared with each other in 2 × 2 pairs. The yeast RNA method had the greatest estimate (1.03 g of MCP) compared to the other methods. Comparative analyses of the different methods presented the lowest variability data for PCRM with SD = 0.07. In conclusion, the four evaluated methods provided different MCP estimates. However, for precise MCP determination, we recommend the PCRM method, with FCM as a reliable alternative. These methods showed lower SD due to better control and primer specificity.

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技术说明：不同的分析方法定量了哺乳山羊瘤胃食糜微生物粗蛋白质池

我们评估了用代乳剂（MR）喂养的断奶前奶山羊皱胃样品中微生物粗蛋白（MCP）池的量化方法。选取平均年龄为2岁 ± 1日龄、平均体重为3.834 ± 0.612 kg的萨宁和瑞士高山山羊公山羊60只。山羊孩子接受不同的流质饮食，分为低营养计划（LNP）；提供1升MR/山羊/天)和高营养计划(HNP；2升MR/山羊/天)。所有山羊幼崽在喂养后的6个时间点收获45天。对皱胃内容物进行取样，以确定微生物池并估计MCP。MCP的定量采用4种方法：酵母RNA法1、Bradford Blue法2 （BBM）、流式细胞术法3 (FCM)；聚合酶链反应-方法4 (PCRM；本研究提出的方法)。4种方法进行2对 × 的相互比较。与其他方法相比，酵母RNA法的MCP估计值最高（1.03 g）。不同方法的对比分析显示，PCRM的变异性最低，SD = 0.07。总之，四种评估方法提供了不同的MCP估计值。然而，为了精确测定MCP，我们推荐PCRM方法，FCM作为可靠的替代方法。由于对照和引物特异性较好，这些方法的SD值较低。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Animal Feed Science and Technology 农林科学-奶制品与动物科学

CiteScore

6.00

自引率

6.20%

发文量

266

审稿时长

3 months

期刊介绍： Animal Feed Science and Technology is a unique journal publishing scientific papers of international interest focusing on animal feeds and their feeding. Papers describing research on feed for ruminants and non-ruminants, including poultry, horses, companion animals and aquatic animals, are welcome. The journal covers the following areas: Nutritive value of feeds (e.g., assessment, improvement) Methods of conserving and processing feeds that affect their nutritional value Agronomic and climatic factors influencing the nutritive value of feeds Utilization of feeds and the improvement of such Metabolic, production, reproduction and health responses, as well as potential environmental impacts, of diet inputs and feed technologies (e.g., feeds, feed additives, feed components, mycotoxins) Mathematical models relating directly to animal-feed interactions Analytical and experimental methods for feed evaluation Environmental impacts of feed technologies in animal production.