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Bacillus pumilus 315 with probiotic effect and its application

A technology of Bacillus pumilus and bacteria agent, applied in the field of microbiology, can solve the problems of less research and application, and achieve the effects of enhancing immune function, reducing feed-to-meat ratio, and good probiotic properties

Active Publication Date: 2017-09-29
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research and application of Bacillus pumilus as a probiotic preparation or feed additive is less, especially in the aspects of replacing antibiotics in feeding and reducing environmental pollution.

Method used

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  • Bacillus pumilus 315 with probiotic effect and its application
  • Bacillus pumilus 315 with probiotic effect and its application
  • Bacillus pumilus 315 with probiotic effect and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 The isolation and identification of 315 (CGMCC NO.8878) of Bacillus pumilus

[0032] The soil near the pig farm was collected and diluted in saline at a ratio of 1:10. Take 100 μL of the diluted solution and spread it on the nutrient broth solid medium, and incubate at 37°C for 24h. Stir the single colony that forms wrinkles on the surface of the colony and separate it by streaking on the solid medium of the nutrient broth. Repeat the process of streaking and separation for 3 rounds to purify the strain. 34 strains were isolated by this method, named 301-334 respectively.

[0033] Bacillus pumilus with antibacterial activity was screened by agar diffusion Oxford cup method. After adding agar to the broth medium, dissolve it evenly, sterilize under high pressure, cool to about 50°C, and add 20 μL of freshly cultured indicator bacteria (10 8 CFU / mL), mix well and pour into a Petri dish to cool. Use sterilized tweezers to gently put the sterile Oxford cup int...

Embodiment 2

[0040] Example 2 Determination of acid and bile salt resistance of Bacillus pumilus 315

[0041] To determine the growth state of Bacillus pumilus 315 in low pH, adjust the nutrient broth medium with 1mol / L hydrochloric acid to have different pH gradients, which are 2.0, 3.0, 4.0, 5.0, 6.0 and 6.4, respectively. Bacillus pumilus 315 culture solution was inoculated in nutrient broth medium with different pH values ​​at 1% (v / v), cultured at 37°C for 24 hours, and the OD was measured 600nm value. Test results such as figure 1 As shown, the survival rate of Bacillus pumilus 315 in the medium with pH = 4.0 is 68%, indicating that Bacillus pumilus 315 can maintain a certain number of viable bacteria in a low-acid environment.

[0042] Pig bile salt was added to the nutrient broth medium to make the concentrations respectively 0% (control), 0.15%, 0.3% and 0.6%. Bacillus pumilus 315 was inoculated in the above-mentioned nutrient broth medium with different concentrations, culture...

Embodiment 3

[0046] Example 3 Determination of tolerance of Bacillus pumilus 315 against artificial gastrointestinal fluid

[0047] Preparation of artificial gastric juice: take 16.4mL of 9.5%-10.5% hydrochloric acid, dilute with water, and make the pH value reach 3.0. Add 1.0g of pepsin to every 100mL of the solution, mix well, and filter with a 0.2μm sterile filter.

[0048] Preparation of artificial intestinal juice: Weigh 6.8g of potassium dihydrogen phosphate, add 500mL of water to dissolve, adjust the pH value to 6.8 with 0.4% sodium hydroxide solution, add water to dilute to 1000mL, add 1.0g of trypsin to every 100mL of liquid, mix well, Filter through a 0.2 μm sterile filter.

[0049] The Bacillus pumilus 315 culture solution was inoculated in the artificial gastric juice and artificial intestinal juice respectively, cultured on a shaker at 37°C and 140r / min, and the plate counts of viable bacteria were carried out at 0, 30 and 180 minutes respectively. The results are shown in T...

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Abstract

The invention discloses a Bacillus pumilus 315 with a preservation number of CGMCC NO.8878. Bacillus pumilus 315 has the characteristics of improving feed utilization, promoting digestion and absorption of nutrients in feed; enhancing animal immune function, increasing daily weight gain, reducing feed-to-meat ratio; no pollution, no residue, and biological environmental protection. The bacillus pumilus of the invention can be used as a novel probiotic additive and widely used in feed.

Description

technical field [0001] The invention belongs to the field of microbiology, and in particular relates to a bacillus pumilus 315 with probiotic effect and its application. Background technique [0002] With the rapid development of animal husbandry in my country, the contradictions between quantity and quality, production and environment, and benefit and resources are becoming more and more prominent. In particular, the pollution caused by livestock and poultry excrement to the environment is becoming more and more serious. A large number of harmful substances discharge make the BOD (biochemical oxygen demand) and COD (chemical oxygen demand) values ​​​​in the pollutants of livestock and poultry farms rise sharply, and it has also become some Potential triggers for disease development and spread. [0003] On the other hand, food safety issues are attracting more and more attention. Among the many causes of food safety problems, one of the important reasons is the use of antib...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/742A61P31/04A23K10/18C12R1/07
CPCA61K35/742C12N1/20C12N1/205C12R2001/07Y02A50/30
Inventor 刁其玉张乃锋祁敏丽
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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