Separation method of total microbes in porcine gastrointestinal contents and manure

Abstract

The invention discloses a gradient centrifugation separation method of total microbes in porcine gastrointestinal contents and manure. The gradient centrifugation separation method comprises the following steps: A, performing reagent preparation of a separation buffer solution; B, performing an experimental method as follows: (1) selecting the porcine gastrointestinal contents and manure and aseptically collecting contents and manure in stomach, jejunum, ileum, cecum and colon; (2) respectively weighing the weight of the contents and manure, placing into a sterile centrifugal tube, centrifuging and discarding supernatant; (3) adding the separation buffer solution into the centrifugal tube, shaking up on a vortex mixer and centrifuging; (4) taking out the centrifugal tube, pouring supernatant fluid into the centrifugal tube, centrifuging the centrifugal tube loaded with the supernatant fluid and discarding the supernatant; (5) weighing the separation buffer solution, adding into a centrifugal tube a, shaking up on the vortex mixer, centrifuging and discarding the supernatant; (6) respectively repeating the step 3, the step 4 and the step 5 twice; (7) pouring away the supernatant fluid, transferring thalli into a 2ml centrifugal tube and preserving at the temperature of -70 DEG C for determining the dry weight of the thalli and a microbial flora. The method is easy, simple and convenient to operate and the cost is low.

Description

technical field [0001] The invention relates to the technical field of microorganism separation, and more specifically relates to a gradient centrifugation method for pig gastrointestinal tract contents and total amount of feces microorganisms. Background technique [0002] The microbial flora in the pig gastrointestinal tract is complex and large in number. The metabolism degree and destination of nitrogen nutrients by microorganisms in the gastrointestinal tract directly affect the body's utilization of dietary nitrogen nutrients and nitrogen discharge. absorb. The study of microbial flora, amino acid composition of microbial cells excreted by endogenous amino acids, and microbial enzyme activities is very important for analyzing the metabolic mechanism of intestinal microorganisms and nitrogen nutrients. The isolation of the total microbial population in the pig gastrointestinal tract is a prerequisite for the study of microflora and the determination of microbial enzym...

AI Technical Summary

Problems solved by technology

Fecal microbial nitrogen and amino acids are important components of endogenous nitrogen and amino acids. The separation of total fecal microbial nitrogen and amino acids is very important for understanding the excretion of fecal microbial nitrogen and amino acids. However, there is no unified separation standard at home and abroad.

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