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Strain, bacterium agent and application

A technology of bacterial strains and inoculants, applied in the field of microorganisms, can solve the problems of low number of viable bacteria in products, difficult to achieve probiotic effects, short shelf life, etc., to achieve excellent probiotic effects, protect the immune system of animals and the effect of gastrointestinal health

Active Publication Date: 2019-04-26
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, probiotics such as Lactobacillus and Bifidobacterium have poor tolerance to adverse environments, resulting in a small number of viable bacteria and a short shelf life, making it difficult to achieve a probiotic effect in the host.

Method used

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  • Strain, bacterium agent and application
  • Strain, bacterium agent and application
  • Strain, bacterium agent and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 (isolation and screening bacterial strain)

[0023] A free-range healthy black-bone chicken was purchased from a farmer in Baitan Village, Xigobi Town, Shawan County, Tacheng Prefecture, Xinjiang. It was fasted for 12 hours (drinking water freely during the period) and then sacrificed by venous bloodletting. Evenly, weigh 10.0g and dissolve in 90mL sterile saline, shake evenly, and bathe in 85°C water bath for 30min; adopt gradient dilution coating method on LB solid medium, and incubate at 37°C for 24h. Preliminary screening was carried out based on phenotypic differences such as colony color, size, and shape. After continuous transfer and streak culture for 3 times, transfer to an inclined test tube and refrigerate at 4°C; pick a single colony and inoculate it into a liquid medium at 37°C and 150r / min Shake enrichment for 18h, centrifuge at 5600r / min for 10min, discard the supernatant, add fresh medium to resuspend, supplement 25% sterilized glycerol and ...

Embodiment 2

[0024] Embodiment 2 (identify bacteriostasis)

[0025] Detect the bacteriostasis of the isolated strains: adjust the concentration of the enriched bacterial solution to (2.0-2.5)×10 8 CFU / mL, pipette 150 μL on LB solid medium and evenly spread it, and incubate it upside down at 37°C for 24 hours; use a puncher to punch out a 6mm-diameter bacterial cake for use; spread the indicator bacteria on the corresponding medium at the same concentration, Place 3 copies of the bacteria cakes on the surface of the indicator bacteria equidistantly, and use LB agar block as a blank control, incubate at a suitable temperature for 18 hours, and measure the diameter of the inhibition zone; the inhibition diameter of strain CCTCC LTNo.1 is shown in figure 2 ; indicator bacteria Escherichia coli EPEC O127:K63CICC 10411 (enteropathogenic Escherichia coli), Escherichiacoli ETEC O78:K80CICC 10421 (enterotoxigenic Escherichia coli), Escherichia coli EHEC O157:H7 CICC21530 (enteropathogenic Escheric...

Embodiment 3

[0026] Embodiment 3 (identification bacterial strain)

[0027] 1. Colony and cell morphology: Streak the strain LTNo.1 on LB agar medium, culture at 37°C for 24 hours, the colony shape is irregular, the edge is off-white and smooth, the middle part is white and raised, and the diameter of the colony is 3-5mm; Gram staining was positive, the cells were short rod-shaped, and the colonies and cell shapes were seen in image 3 .

[0028] 2. Physiological and biochemical identification: Physiological and biochemical identification of strain LTNo.1 was carried out referring to the method in the "Common Bacterial System Identification Manual", and the results are shown in Table 1.

[0029] Table 1. Physiological and biochemical characteristics of strain LTNo.1

[0030]

[0031] Note: "+": Positive, "-": Negative.

[0032] 3. Molecular identification: the molecular biology method of 16S rRNA gene sequencing was used to identify the above-mentioned bacterial strain; the 16S rRNA ...

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Abstract

The invention discloses a strain, a bacterium agent and application and relates to the technical field of microorganisms. The strain is bacillus subtilis; the strain is preserved in China Center for Type Culture Collection (CCTCC), the preservation number is CCTCC NO: M 2018398 and the preservation date is June 25, 2018. The strain provided by the invention can be used for inhibiting 5 types of pathogenic microorganisms and is sensitive to 15 types of antibiotics; the strain has a non-specific adhesion potential, has tolerance to acid and bile salt, has the property of producing pectinase andcellulase and also has a haemolysis-free property; the strain provided by the invention has an excellent probiotic effect, can be used for controlling and regulating the balance of intestinal flora ofanimals and is used for preparing an animal feed additive to protect the immune system and gastrointestinal health of the animals.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bacterial strain, bacterial agent and application. Background technique [0002] The conflict between the world's growing population and dwindling available land resources has become increasingly acute. In order to meet people's demand for animal protein, intensive animal farming methods have continued to develop over the past few decades. During this period, antibiotics made a huge contribution to intensive farming due to their use in the treatment of infectious diseases in animals. At the same time, antibiotics have been widely used as animal growth promoters since their discovery, which has caused the accumulation and spread of multi-drug resistance genes in pathogenic microorganisms, thus seriously threatening the health of humans and animals. Therefore, the European Union banned all antibiotics as growth promoters in feed in 2006. With the prohibition of antibioti...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/18C12R1/125
CPCA23K10/18C12N1/20C12N1/205C12R2001/125
Inventor 倪永清罗宝龙张艳田会丽剡文莉张亚川
Owner SHIHEZI UNIVERSITY
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