Method for screening functional feed additives for piglets

A technology of feed additives and screening methods, which is applied in the direction of biochemical equipment and methods, analytical materials, and microbial determination/inspection, etc., can solve the problems of inaccurate evaluation of the effect of candidate nutrients, lack of screening methods, lack of real-time monitoring, etc., to achieve Improving the mitochondrial respiratory function of intestinal cells, with high accuracy and fast speed

Pending Publication Date: 2018-04-20
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is a lack of convenient screening methods for feed additives, and it often occurs that candidate nutrients are directly tested on animals without screening, often resulting in a great waste of manpower and material resources due to poor test results
Traditional screening methods include in vitro antibacterial test, in vitro cell growth test, etc., lack of real-time monitoring and analysis of cell-related functions, and cannot accurately evaluate the effect of candidate nutrients

Method used

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  • Method for screening functional feed additives for piglets
  • Method for screening functional feed additives for piglets
  • Method for screening functional feed additives for piglets

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Example 1 Real-time monitoring of the growth of porcine small intestinal cells IPEC-1

[0103] In this example, all liquids were sterilized through a 0.2 μM filter.

[0104] (1) The porcine intestinal cells IPEC-1 were cultured in vitro.

[0105] (2) Preliminary preparation: the 75cm 2 IPEC-1 cells with a growth abundance of 80% in the culture flask were washed twice with 10ml PBS (phosphate buffered solution), i.e., phosphate buffered saline, and then digested by adding 200μl Trypsin (Parenzyme) for 5 minutes. Observe the cell state, digest until the cells are dispersed, and then stop the digestion with 10ml DMEM / F12 medium containing 10% fetal bovine serum;

[0106] Transfer the DMEM / F12 medium loaded with porcine small intestinal cells IPEC-1 to a 15ml centrifuge tube, and centrifuge at room temperature at 1000 rpm for 5 minutes.

[0107] The supernatant in the DMEM / F12 medium loaded with porcine small intestinal cells IPEC-1 after centrifugation was sucked off, a...

Embodiment 2

[0117] Example 2 Determination of the mitochondrial respiratory function of pig small intestinal cells IPEC-1

[0118] (1) The porcine intestinal cells IPEC-1 were cultured in vitro.

[0119] (2) Preliminary preparation: the 75cm 2 IPEC-1 cells with a growth abundance of 80%-90% in the culture flask were washed twice with 10ml PBS (phosphate buffered solution), i.e. phosphate buffered saline, and then digested by adding 200 μl of trypsin (Parenzyme) for 5 minutes. Use a microscope to observe the cell state, digest until the cells are dispersed, and then stop the digestion with 10ml DMEM / F12 medium containing 10% fetal bovine serum;

[0120] Transfer the DMEM / F12 medium loaded with porcine small intestinal cells IPEC-1 to a 15ml centrifuge tube, and centrifuge at room temperature at 1000 rpm for 5 minutes.

[0121] The supernatant in the DMEM / F12 medium loaded with porcine small intestinal cells IPEC-1 after centrifugation was sucked off, and the DMEM / F12 medium that added 1 ...

example 1

[0136] Examples 1 and 2 take glutamine (Gln) as an example as a candidate nutrient for functional feed additives for specific analysis.

[0137] Such as figure 1 Shown, be divided into three groups 0mM Gln group (control group), 0.5mM Gln group (treatment group), 2mM Gln group (treatment group), by figure 1 It can be seen that after adding different concentrations of Gln, the vitality and quantity of pig small intestinal cells IPEC-1 were significantly improved, especially the effect of promoting cell growth and enhancing cell viability in the 2mM Gln group was better than that in the 0.5mM Gln group;

[0138] Such as figure 2 As shown, divided into 2 groups of control group (0mM Gln group) and treatment group (2mM Gln group), from figure 2 It can be seen from the figure that after adding 2mM Gln to the test solution, the respiratory oxygen consumption rate OCR of the pig small intestine cell line IPEC-1 is rapidly increased (<8min). The results of the calculation of the m...

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Abstract

The invention discloses a method for screening functional feed additives for piglets. The method for screening the functional feed additives for the piglets comprises the following steps: culturing porcine intestinal cells IPEC-1 in vitro; adding candidate nutrients from a culture medium of the porcine intestinal cells IPEC-1, and carrying out real-time observation on cell growth state and cell mitochondrial respiratory function indexes; screening the functional feed additives for the piglets from the candidate nutrients according to the cell growth state and the cell mitochondrial respiratoryfunction indexes. The invention aims at providing the method for screening the functional feed additives for the piglets by carrying out cell monitoring in real time and combining a cellular mitochondrial respiratory metabolism function as the assessment index.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for screening functional feed additives for piglets. Background technique [0002] For a long time, the addition of antibiotic growth-promoting agents in feed has provided humans with a large number of economical livestock and poultry products, but it has also produced drug resistance and residue problems, seriously affecting the ecological environment and food safety. An important problem in the current development of the feed industry and the breeding industry is how to reduce the use of antibiotics in feed without affecting the application effect of feed products and the production efficiency of pig and poultry farming. Finding safe and efficient feed additives to replace feed antibiotics is a major scientific and technological problem that my country's feed industry and aquaculture industry urgently need to solve. [0003] At present, there is a lack of convenient scree...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04
CPCG01N33/5044
Inventor 易丹侯永清丁斌鹰吴涛
Owner WUHAN POLYTECHNIC UNIVERSITY
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