Detection method of lactic acid bacteria flora changes in biological feed fermentation process

A technology for biological feed and fermentation process, applied in the field of molecular biology, can solve the problems of few reports of changes in bacterial flora, lack of effective methods for the identification of bacteria with similar growth and colonies, etc., and achieves short detection period, simple operation and repeatability. Good results

Pending Publication Date: 2019-08-02
浙江康星生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the research on fermented feed is mostly focused on the change of feed nutrient composition and application effect, but there are few reports on the changes of relevant flora in fermented feed, and there is no effective method for the identification of bacterial colonies and bacteria with similar growth potential.

Method used

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  • Detection method of lactic acid bacteria flora changes in biological feed fermentation process
  • Detection method of lactic acid bacteria flora changes in biological feed fermentation process
  • Detection method of lactic acid bacteria flora changes in biological feed fermentation process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1: Quantitative analysis of changes in the number of lactic acid bacteria flora during the step-by-step fermentation of biological feed

[0052] The raw materials are 80g of corn stalks and 20g of bran, a total of 100g, the water content is 40%, the added aerobic bacteria are Bacillus subtilis and Saccharomyces cerevisiae, and the added lactic acid bacteria are the same amount of Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium longum bacillus, Lactobacillus casei and Lactobacillus rhamnosus, the addition of aerobic bacteria and lactic acid bacteria are all 5% of the raw material. The fermentation method adopted is step-by-step fermentation, that is, aerobic bacteria are added to aerobic fermentation for 12 hours, and then lactic acid bacteria are added, and immediately sealed for anaerobic fermentation. A total of 6 samples were set up, and each sample was replicated 3 times. The DNA of the 0d, 1d, 2d, 3d, 5d, 7d, 10d, 15d, 20d, 25d and 30d s...

Embodiment 2

[0056] Example 2: Quantitative analysis of changes in the number of lactic acid bacteria flora during the simultaneous fermentation of biological feed

[0057] The same conditions as in Example 1 were adopted, but the step-by-step fermentation was changed to synchronous fermentation, that is, aerobic bacteria and anaerobic bacteria were added at the same time and then directly packaged for fermentation.

[0058] For specific results, see Image 6 , 7 , 8, 9 and 10. From the test results, it can be seen that the step-by-step fermentation of aerobic bacteria and anaerobic probiotics is compared with the synchronous fermentation after adding aerobic bacteria and anaerobic probiotics at the same time. From the sample F mixed with five kinds of bacteria, the measured value of simultaneous fermentation The bacterial counts of the five kinds of bacteria are all larger than those measured by the step-by-step fermentation. For example, the number of Lactobacillus plantarum in sample ...

Embodiment 3

[0059] Embodiment 3: molecular biology experiment

[0060] 3.1 Culture of bacteria and extraction of genomic DNA

[0061] Fully mix the lyophilized powder of the standard strain with an appropriate amount of sterile saline, inoculate it in PDA solid medium, and then place it in a constant temperature incubator at 37°C for 48 hours, then pick a single colony with a sterile inoculation loop and inoculate it into MRS In the culture medium, culture anaerobically at 37°C to activate the strain until the OD of the strain 600 The value is 0.8.

[0062] The cultured fresh bacteria of the standard strains were collected, and the bacterial DNA was extracted using the Bacterial Genome Extraction Kit (Shanghai Sangong). For the specific operation steps, see the kit instruction manual.

[0063] 3.2 Design and synthesis of primers

[0064] Primer5.0 primer design software was used to design specific primers for five lactic acid bacteria. The obtained primers are shown in Table 1. The obt...

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Abstract

The invention discloses a detection method of lactic acid bacteria flora changes in a biological feed fermentation process, which comprises the flora number changes of lactobacillus plantarum, lactobacillus acidophilus, bifidobacterium longum, lactobacillus casei and lactobacillus rhamnosus, The specific, sensitive and low-cost detection method is provided for determination of the amount of anaerobic probiotic bacteria in the biological feed fermentation process, which can simultaneously detect five types of anaerobic probiotic bacteria and understand a flora change rule of the five types of anaerobic probiotic bacteria in the fermentation process, and provides a technical solution for further optimizing the fermented feed.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for detecting the change of lactic acid bacteria flora during the fermentation process of biological feed. Background technique [0002] Microbial fermented feed refers to the addition of lactic acid bacteria, yeast, bacillus and other microorganisms to one or more feed materials under human controllable conditions for aerobic or anaerobic fermentation. During the fermentation process, microbial metabolism Function, eliminate the anti-nutritional factors in the feed raw materials, make full use of the nutrients in the raw materials, so as to obtain a new type, safe, nutritious and easy to digest and absorb biological feed. [0003] The solid feed fermented by lactic acid bacteria can continuously degrade some macromolecular nutrients (such as protein, polysaccharide, fat, etc.) And unknown growth factors, etc., so as to improve the nutritional value of feed....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851C12Q1/06
CPCC12Q1/6851C12Q2531/113C12Q2563/107
Inventor 陈华友李婷婷李继彬崔凤杰齐向辉
Owner 浙江康星生物科技有限公司
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