Swine-origin lactobacillus reuteri and prepared solid probiotics

A technology of porcine-derived Lactobacillus reuteri and probiotics, applied in bacteria, microorganisms, microorganisms and other directions, can solve the problems of high degree of protein denaturation, low cost of unfavorable culture medium, low digestion and utilization rate, etc. The effect of reducing the cost of green pig breeding and high viable count

Inactive Publication Date: 2017-08-15
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it still meets the basic requirements of unfavorable industrial production medium and low cost
[0007] Soybean meal is a by-product of oil extraction with organic solvents. Although the protein content of soybean meal is 44%-48%, in view of...

Method used

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  • Swine-origin lactobacillus reuteri and prepared solid probiotics
  • Swine-origin lactobacillus reuteri and prepared solid probiotics
  • Swine-origin lactobacillus reuteri and prepared solid probiotics

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Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0022] Specific embodiment one: the separation and screening of bacterial strain

[0023] Isolation and screening of acid producing strains:

[0024] Take an appropriate amount of fresh manure from healthy sows and add it to a sterile container, seal it immediately, and bring it back to the laboratory under ice-cold conditions. Under sterile conditions, take a little of the above-mentioned fresh feces and add them to a sterile water conical flask with glass beads, shake well, take 1 mL and add 9 mL of sterile water, mix well, and sequentially dilute to 10-6, and take each gradient sample solution Pour 1mL into a sterilized empty plate, then pour 15mL-20mL / plate of the separation medium that has been sterilized and cooled to about 45°C and melted, shake it horizontally, put it upside down in a constant temperature incubator after solidification, and keep it at 37°C for 2-3 days . Pick a single colony that turns red around it and puncture it into the sterilized semi-solid MRS ...

specific Embodiment approach 2

[0032] Specific implementation mode two: identification of strain morphology, physiology and biochemistry

[0033] Morphological, physiological and biochemical tests and API50CH tests of carbohydrate acid production were carried out on the isolated strains, and the results are shown in Table 1 and Table 2, respectively. Gram-positive strains, see appendix image 3 . According to "Bergey's Handbook of Systematic Bacteriology", "Handbook of Common Bacterial System Identification" and relevant research papers in International Journal of Systematic and Evolutionary Microbiology, it was initially identified as Lactobacillus reuteri.

[0034] Table 1 Physiological and biochemical test results of strain Lactobacillus reuteri YSJL-12

[0035]

[0036] Table 2 Results of carbohydrate fermentation acid production (API50CH) of strain Lactobacillus reuteri YSJL-12

[0037]

specific Embodiment approach 3

[0038] Specific embodiment three: 16SrRNA gene sequence analysis method to identify bacterial strains

[0039] Strain genome extraction: Extract the genome of the isolated strain according to the TIANamp Bacteria DNA Kit Bacterial Genome DNA Extraction Kit, and perform 1% agarose gel electrophoresis detection.

[0040] 16SrRNA gene PCR amplification: primer 1 799F 5'-AACAGGATTAGATACCCTG-3' and primer 2 1492R5'-GGTTACCTTGTTACGACTT-3'. Reaction system (25uL): 10×PCR buffer 2.5uL, dNTP (2.5mM) (TaKaRa) 2uL, 799F (10pmol / uL) (invitrogen) 0.1uL, 1492R (10pmol / uL) (invitrogen) 0.1uL, Taq polymerase (5U / uL) (TaKaRa) 0.125uL, Templet (extracted DNA) 0.5uL, ddH2O to 25uL. Reaction conditions: pre-denaturation at 94°C for 5min, denaturation at 94°C for 1min, refolding at 52°C for 1min for 30cycles, extension at 72°C for 1min, and final extension at 72°C for 10min. After the PCR reaction, use 1% agarose gel electrophoresis to detect the PCR product .

[0041] 16SrRNA gene amplified fr...

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Abstract

The invention relates to swine-origin lactobacillus reuteri and prepared solid probiotics. The bacterial strain is separated from fresh night soil of healthy live pig, and is indentified as Lactobacillus reuteri YSJL-12 through a morphological biochemistry and molecular biology method, and a preservation number is CGMCC No. 9062. The bacterial strain inhibits Listeria monocytogenes CICC21633 to positive. The prepared solid probiotics contain soybean meal and active Lactobacillus reuteri YSJL-12, can substitute aureomycin for preventing and treating diarrhoea while added in a live pig basic feed, and the feed-gain ratio of the breeding pork pig at age in days of 28-168 is 2.61. The bacterial strain has the characteristics of strong host specificity, large live bacteria quantity, low cost and high cost performance, added value of high temperature soybean meal is increased, and the green live pig culture cost is reduced.

Description

technical field [0001] The invention relates to an isolated strain of Lactobacillus reuteri from pigs and a feed probiotic agent prepared by using the strain which can replace aureomycin, and belongs to the field of feed additives. Background technique [0002] The addition of antibiotics to feed has become a common fact in the aquaculture industry in the world today. From the perspective of short-term effects, the feed with antibiotics can indeed inhibit the diarrhea of ​​fed animals and kill pathogenic bacteria in the intestinal tract, but at the same time destroy the normal intestinal bacteria of animals. group balance. From a long-term perspective, long-term use of antibiotics will cause the pathogenic bacteria in the intestinal tract to develop drug resistance, resulting in a gradual increase in the dose of antibiotics to kill the pathogenic bacteria, increasing the risk of antibiotic residues, and eventually leading to no medicine for human beings apply. Therefore, i...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/18A23K50/30C12R1/225
CPCC12N1/20C12N1/205C12R2001/225
Inventor 徐速江连洲孙慧许岩穆莹
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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