Bacillus subtilis and application thereof to livestock breeding

A technology of Bacillus subtilis and livestock and poultry feed, which is applied in the fields of application, bacteria, and animal husbandry. It can solve the problems of large differences in probiotic performance and fewer reports on the overall performance of strains, so as to promote the development of immune organs and improve production performance. , the effect of promoting reproduction

Active Publication Date: 2014-02-05
山东蔚蓝生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the probiotic properties of different Bacillus strains vary widely
At present, most studies use a single performance as the screening index, such as simply examining the antibacterial activity or enzyme production ability, etc. There are few reports on the consideration of the overall performance of the strain

Method used

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  • Bacillus subtilis and application thereof to livestock breeding
  • Bacillus subtilis and application thereof to livestock breeding
  • Bacillus subtilis and application thereof to livestock breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1 Isolation and Identification of Bacillus subtilis WL-04

[0014] 1. Isolation and screening of Bacillus

[0015] Add the sludge collected from the Pingdu Broiler Farm in Qingdao City into sterile water at a ratio of 1:10, shake and mix to prepare a suspension, and treat it in a water bath at 80°C for 10 minutes to kill most of the non-heat-resistant Bacillus with good heat resistance was preliminarily obtained.

[0016] 2. Screening of Bacillus with ability to inhibit pathogenic bacteria and high enzyme production

[0017] 1) Preparation of bacillus bacteria liquid: inoculate the isolated bacillus into nutrient broth respectively, shake and cultivate for 24 hours at 37°C and 200rpm;

[0018] 2) In vitro antibacterial test - micro broth method: Centrifuge the bacillus liquid cultured for 24 hours, collect the supernatant, take a sterile 96-well culture plate, add the bacillus supernatant and pathogenic bacteria liquid (used in the experiment) at a ratio of 1:...

Embodiment 2

[0029] Embodiment 2: Bacillus subtilis WL-04 inhibits the analysis of zoonotic pathogens

[0030] 1. Cultivation of indicator pathogenic bacteria: Randomly select pathogenic bacteria isolated from the intestinal tract of sick chickens and pigs, including Escherichia coli, Salmonella, and Clostridium perfringens, a total of 5 strains (gifted by the Preventive Veterinary Laboratory of Qingdao Agricultural University), of which Escherichia coli, Salmonella was inoculated in nutrient broth, 37°C, 200rpm, cultured for 15h; Clostridium perfringens was inoculated in blister medium, 37°C, cultured for 15h;

[0031] 2. Preparation of Bacillus subtilis WL-04 bacterial liquid: pick a single colony, inoculate it in nutrient broth, culture at 37°C, 200rpm, for 24 hours;

[0032] 3. Antibacterial test in vitro - trace broth method: centrifuge the bacillus liquid cultured for 24 hours, collect the supernatant, take a sterile 96-well culture plate, add the bacillus supernatant and pathogenic ...

Embodiment 3

[0041] Embodiment 3: Bacillus subtilis WL-04 produces enzyme performance analysis

[0042] 1. Prepare Bacillus subtilis WL-04 bacterial liquid: pick a single colony, inoculate it in nutrient broth, culture at 37°C, 200rpm, for 24 hours;

[0043] 2. Obtain the fermentation supernatant: centrifuge the bacillus liquid cultured for 24 hours at 10,000 rpm for 10 minutes, and collect the supernatant;

[0044] 3. Enzyme production: α-amylase activity detection (GB / T24401-2009) and neutral protease activity detection (GB / T28715-2012), a total of 3 batches of determination were carried out, the results are shown in Table 3;

[0045] Table 3: Analysis of enzyme production by Bacillus subtilis WL-04

[0046]

[0047]

[0048] After three batches of repeated experiments and the analysis of enzyme activity, the enzyme production of Bacillus subtilis WL-04 was relatively stable, and the average activity of α-amylase was 22U / mL; the average activity of neutral protease was 170U / mL. M...

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Abstract

The invention aims to provide bacillus subtilis WL-04 and application thereof to livestock breeding. The preservation number of the bacillus subtilis WL-04 is CCTCC NO:M 2013343. The bacillus subtilis WL-04 can be used for remarkably inhibiting the growth of Escherichia coli, salmonella, clostridium pefringens and other conditional pathogenic bacteria of the intestinal canal of livestock, and particularly, the bacteriostasis rate of the bacillus subtilis WL-04 to Escherichia coli and clostridium perfingens is above 75%; the enzyme production is stable; the average activity of alpha-amylase is 22U/mL, and the average activity of neutral protease is 170U/mL; the reproduction of probiotics of intestinal canal can be promoted; high temperature for pelletizing can be endured; the influence of inverse environments with low-pH gastric fluid and high-concentration cholate is endured; the production performance of breeding animals can be effectively improved; immune organ development can be promoted. The bacillus subtilis WL-04 can be widely applied to livestock feeds as a feed additive.

Description

technical field [0001] The invention belongs to the technical field of microbial screening and application, and in particular relates to a bacillus subtilis WL-04 strain and its application as a feed additive. Background technique [0002] Nowadays, with the rapid development of large-scale and intensive breeding, there have been problems such as excessive stocking density and poor ventilation in the breeding house, and the quality of the breeding environment has declined. Various digestive tract diseases of livestock and poultry caused by conditional pathogenic bacteria have a great impact on the environment. Breeding problems are also becoming more and more prominent, such as colibacillosis, salmonellosis, and necrotic enteritis caused by Clostridium perfringens. In order to achieve the purpose of preventing diseases or promoting animal growth, a large number of antibiotics are used in multiple links of livestock and poultry production. The long-term use and abuse of anti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K1/16C12R1/125
Inventor 周英俊孔垂彬孟祥余
Owner 山东蔚蓝生物科技有限公司
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