Method for evaluating activity of bio-feed

A technology of activity and feed, applied in the field of activity evaluation of biological feed, can solve the problems of evaluating active feed and lack of active feed evaluation system, and achieve the effects of improving growth performance, preventing sow constipation, and treating piglet diarrhea.

Inactive Publication Date: 2017-04-26
深圳市前海金卓生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] 2. Lack of active feed evaluation system
[0011] Up to now, there is no method to evaluate the activity of active feed by using metagenomic technology to detect positive o...

Method used

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  • Method for evaluating activity of bio-feed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Method Example 1 Establishment of a mouse model transplanted with pig-derived flora

[0063] Sterile C57BL / 6J mice (provided by the Experimental Animal Center of the Third Military Medical University) were kept in a sterile isolator in the Teaching and Research Office of Experimental Zoology, Basic Department, Third Military Medical University, and the breeding environment: 20-24°C, humidity 40%-70% , strict 12h light 12h dark. The feed was from the co60 irradiated rat growth and breeding feed sold by Beijing Keao Xieli Feed Co., Ltd. The litter was sterilized by irradiation with Co-60γ50kGy rays. Rat cages, drinking water and drinking bottles were all sterilized by high temperature and high pressure (121°C, 60min ). Thirty 4-week-old sterile C57BL / 6J mice were fed with 0.1 mL of pig feces suspension (from healthy young pigs, without gastrointestinal diseases, normal diet and living habits, and not fed 2 months before the start of the experiment) The first fresh feces...

Embodiment 2

[0064] Method Example 2 Ex vivo chemostat model operation

[0065] Inject 500ml of sterilized intestinal flora culture medium into the fermenter of the isolated chemostat model (assemble the model according to the instructions of the application (patent) No. 200710028346.2), seal it with wax, and make the whole system in a sterile, airtight environment . Turn on the peristaltic pump connected to the fresh medium for 35ml.h -1 Flow rate operation, conveying fresh medium to continuously replenish nutrients, and waste liquid is removed at the same flow rate to ensure a constant culture medium. Accurately weigh pig feces samples (collected from healthy young pigs, without gastrointestinal diseases, normal diet and living habits, and pigs that have not been fed any antibiotics and probiotics 2 months before the start of the experiment, the first time in the morning fresh feces) 5g, diluted 5 times with sterilized 0.85% normal saline, fully homogenized and filtered, got 3 equal sa...

Embodiment 316

[0066] Method Example 3 16S rDNA sequencing

[0067] Extract the total DNA of the sample, using 16S rDNA gene V 3 -V 4 The variable region is used as a target for amplification, universal primer sequence: 341F-806R Forward primer (5'-3'): ACTCCTACGGGRSGCAGCAG (341 F) (SEQ ID NO: 1), reverse primer (5'-3') : GGACTACVVGGGTATCTAATC (806 R) (SEQ ID NO: 2) (referring to Sun, W.et al.Mechanismand Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure.Sci.Rep.6,30237; doi:10.1038 / srep30237(2016).), the amplified PCR product was purified and then sequenced on the machine. The entire sequencing process was completed by Shanghai Ruiyi Biotechnology Co., Ltd., including DNA extraction and primer design, etc., using Illumina HiSeq PE250 for on-machine sequencing. By performing OUT (operational taxonomic unit) cluster analysis on the sequencing results, the species composition of the community, the evolutionary relationship between...

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Abstract

The invention relates to a method for evaluating activity of a bio-feed. Concretely speaking, the invention relates to a method for evaluating activity of the bio-feed by taking intestinal flora as a target object, an aseptic animal is used for detecting the positive or negative fluctuation of the animal intestinal flora for through 16S rDNA sequencing, and the activity of the bio-feed is evaluated. The method can be used for evaluating activity of various bi-feeds, the effect of a detection substance by the intestinal flora can be detected through in-vivo and in-vitro experiment, and the superiority-inferiority of the sample can be reflected through flora fluctuation.

Description

technical field [0001] The invention relates to a method for evaluating the activity of biological feed, in particular to a method for evaluating the activity of biological feed with intestinal flora as a target. Background technique [0002] 1. The situation of banning antibiotics in feed is becoming increasingly severe [0003] The abuse of antibiotics destroys the intestinal flora, seriously endangering the health of animals and humans, and antibiotic residues also endanger food safety. From chemical farming to biological farming will be the development trend, and the theory of active feed is in line with the connotation of biological farming. In the context of increasing attention to food safety and calls for banning antibiotics, the current chemical farming model is becoming more and more unsustainable. The biological breeding model with natural active ingredients such as probiotics and plant extracts as the main feed additive raw materials will be a development trend...

Claims

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Application Information

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Owner 深圳市前海金卓生物技术有限公司
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