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Pig-source lactobacillus mucosae and application thereof

A technology of mucosal lactobacillus and porcine origin, applied in the field of microorganisms, can solve the problems of effect effects and other problems, and achieve the effect of good in vitro probiotic potential

Active Publication Date: 2021-10-19
GANSU AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, the effectiveness of toxoid vaccines to prevent infection with this pathogen is susceptible to its toxoid antigenic composition

Method used

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  • Pig-source lactobacillus mucosae and application thereof
  • Pig-source lactobacillus mucosae and application thereof
  • Pig-source lactobacillus mucosae and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1, Lactobacillus mucosa Lactobacillus mucosae Isolation and identification of

[0029] 1. Isolation and purification of strains

[0030] Take about 0.5g of the feces sample of healthy cooperative pigs and dilute it with 1×DPBS buffer solution to 10 -3 、10 -4 、10 -5 3 gradients. Take 70.00 µL of feces dilutions of different concentrations and smear them on CaCO2 containing vancomycin (sterilized by filtration, final concentration 2.00 mg / mL). 3 - MRS plates were cultured anaerobically at 37°C for 48 h. Pick a single colony that conforms to the morphological characteristics of typical Lactobacillus colonies with obvious calcium dissolution circles, streak culture on the MRS plate, and repeat the purification three times. Select 70 suspected strains that meet the morphological characteristics of Lactobacillus colonies such as milky white, round, smooth edges, and overall plumpness ( figure 1 ).

[0031] The composition of the CaCO3-MRS medium used in the p...

Embodiment 2

[0048] Example 2, Lactobacillus mucosa Lactobacillus mucosae Analysis of biological characteristics

[0049] 1. Growth curve and acid production curve

[0050] activated Lactobacillus mucosa Lactobacillus mucosae Inoculate 1.00% of the inoculum into MRS broth, culture in a shaker at 37°C for 24 h, take samples every 2 h, and measure the absorbance value (OD) of the medium. 600 ) and pH value, the growth curve and acid production curve were plotted with the culture time as the abscissa, and the absorbance value and pH as the ordinate, respectively. The results showed that the growth was relatively slow from 0 to 8 h, the logarithmic growth phase was from 8 to 20 h, and it entered the stable phase and decay phase after 20 h; the pH gradually decreased during the culture process, and the pH value of the MRS broth decreased to Below 4.50 and tends to be stable ( Image 6 ), indicating that it has good acid-producing properties.

[0051] 2. Determination of Acid Resistance

...

Embodiment 3

[0060] Example 3, Lactobacillus mucosa Lactobacillus mucosae Application in the prevention and treatment of pig IPEC-J2 cell injury caused by Clostridium perfringens type C

[0061] 1. Different concentrations of Lactobacillus mucosa Lactobacillus mucosae Effects on the viability of IPEC-J2 cells

[0062] After the IPEC-J2 monolayer cells were rinsed twice with sterile PBS, the following treatments were performed: (1) control group: culture IPEC-J2 cells in 2 mL DMEM / F12 medium; (2) treatment group: 10 7 、10 8 、10 9 CFU / mL Lactobacillus mucosa Lactobacillus mucosae Co-cultured with IPEC-J2 respectively. After 2 h, 4 h, and 6 h of treatment, the supernatant was drawn into a 10 mL sterilized centrifuge tube, and the cells were washed 3 to 5 times with sterile PBS, and then the cells were digested with 300 μL of 0.05% trypsin-EDTA. After terminating the digestion, centrifuge at 309×g for 5 min, discard the supernatant, and resuspend the cells with 500 μL DMEM / F12 basal c...

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Abstract

The invention discloses a pig-source lactobacillus mucosae and an application thereof. The strain is named as lactobacillus mucosae LM410, and the preservation number is CGMCC No.22828. The strain is separated from manure of healthy pigs, and has good acid production, acid resistance, cholate resistance and pathogenic bacterium antagonism. The lactobacillus mucosae LM410 provided by the invention can inhibit the growth of C-type clostridium perfringens, has an obvious protection effect on pig intestine epithelial cell injury caused by the C-type clostridium perfringens, and has a wide development prospect and application value in the aspect of preventing and treating bacterial infectious intestinal diseases of livestock and poultry.

Description

technical field [0001] The invention relates to a cooperative pig-derived mucosal lactobacillus and an application thereof, belonging to the technical field of microbes. Background technique [0002] Piglet diarrhea has always been one of the main diseases that plague pig production, seriously affecting the economic benefits of pig farming. According to reports, the mortality rate of piglets in my country is 15% to 20%, among which the number of piglet deaths caused by diarrhea accounts for about 40% of the total number of deaths, and the direct economic loss to the pig industry is tens of billions of dollars every year. Clostridium perfringens type C, also known as Clostridium welchii type C, is one of the main pathogens causing diarrhea in piglets; its infection mainly occurs in piglets less than 2 weeks old, and has the characteristics of acute onset and short course. The mortality rate of sick piglets reaches more than 70%, which seriously affects the healthy developmen...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P1/12A61P31/04C12R1/225
CPCA61K35/747A61P1/12A61P31/04Y02A40/70
Inventor 杨巧丽张生伟裴利君高小莉王鹏飞滚双宝马艳萍
Owner GANSU AGRI UNIV
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