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Compound preparation containing clostridium butyricum and pediococcus acidilactici as well as preparation method and application of compound preparation

A technology of Pediococcus lactis and Clostridium butyricum, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria used in food preparation, etc., can solve the problems of high production costs of solid products, unstable storage of liquid products, and inability to Large-scale application and other issues to achieve the effect of solving cost and stability problems, reducing bait coefficient, increasing weight gain rate and survival rate

Active Publication Date: 2021-04-06
QINGDAO SHANGDE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main problem encountered by these two kinds of microbial agents involved in the preparation is that the production cost of solid products is still high, which limits the frequency of use; and liquid products cannot be used on a large scale due to unstable storage.

Method used

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  • Compound preparation containing clostridium butyricum and pediococcus acidilactici as well as preparation method and application of compound preparation
  • Compound preparation containing clostridium butyricum and pediococcus acidilactici as well as preparation method and application of compound preparation
  • Compound preparation containing clostridium butyricum and pediococcus acidilactici as well as preparation method and application of compound preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Screening, isolation and identification of Clostridium butyricum GBW-N1

[0034] 1. Isolation, screening and purification of Clostridium butyricum GBW-N1

[0035] The bacterial suspension cultured from the collected samples was serially diluted and spread on tryptone-sulfite-cycloserine (TSC) agar (TSC) medium. After multiple separation and purification, a single colony was obtained, which was named GBW-N1 and preserved.

[0036] The bacterium colony of described bacterial strain GBW-N1 is cultivated on TSC solid medium for 24h figure 1 As shown, its colony diagram is as shown in 1a, round, black, smooth surface, slightly raised in the middle, and irregular edges; the bacterial cell of the bacterial strain GBW-N1 cultured on TSC liquid medium for 48h figure 1 As shown in b, the bacteria are rod-shaped, 1.1-1.2 μm × 2.1-3.6 μm, single arrangement, Gram-positive, spores are fusiform, mesozoic, and the cyst is enlarged.

[0037] 2. Molecular identification of Clostrid...

Embodiment 2

[0048] Example 2: Exploration of co-culture shake flask conditions of Clostridium butyricum and Pediococcus lactis

[0049]Test 1: Configure 220mL MRS culture based on 250mL Shu cattle screw bottle, divide into 3 groups, co-cultivation group: inoculate 1% Clostridium butyricum seed solution and 1% Pediococcus lactis seed solution; control group 1 (CK1): only Inoculate 1% Pediococcus lactis seed liquid; Control group 2 (CK2): only inoculate 1% Clostridium butyricum seed liquid, all three groups are placed in a 37°C incubator for static culture for 20-28 hours. The results (Table 1) show that the co-cultivation of Pediococcus lactis and Clostridium butyricum with MRS general medium for lactic acid bacteria, the growth advantage of Pediococcus lactis is better than that of Clostridium butyricum, and the content of Clostridium butyricum in co-cultivation is less than that of bacteria in single culture. 0.3% of the content, this reason is that Pediococcus lactis grows faster and ca...

Embodiment 3

[0074] Embodiment 3: Clostridium butyricum and Pediococcus lactis co-culture 50L fermenter exploration

[0075] During the cultivation process in the screw bottle, the pH cannot be precisely controlled, and nitrogen pressure cannot be maintained, and a strict anaerobic environment cannot be formed. This has a certain impact on Clostridium butyricum, and the fermentation medium for co-cultivation can only be initially determined. In order to further optimize the process, the optimization test was continued in the 50L fermenter.

[0076] Experiment 1: Inoculate a single colony of Clostridium butyricum GBW-N1 in a Keer flask containing RCM medium, culture at 37°C for 3 days, scrape and wash the bacterial lawn with 0.8% saline as the seed liquid; pick lactic acid tablets A single colony of cocci was inoculated in MRS liquid medium, and expanded at 37°C for 16 hours as a seed solution.

[0077] The formula of the fermentation medium is tryptone 8g / L, beef extract 5g / L, yeast extra...

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Abstract

The invention discloses a compound preparation containing clostridium butyricum and pediococcus acidilactici as well as a preparation method and application of the compound preparation. The compound preparation comprises clostridium butyricum with the bacterium content of 5-6*10<8> CFU / mL and pediococcus acidilactici with the bacterium content of 5.0-6.0*10<9> CFU / mL, wherein the clostridium butyricum adopts clostridium butyricum GBW-N1 with the preservation number of CGMCC No. 14499. The compound preparation is prepared by inoculating two strains in a staggered manner and carrying out fermentation culture in a designed fermentation culture medium for 16-22 hours, and fermentation liquor can be used after being filled. The compound preparation can well improve the weight gain rate, the survival rate and the growth performance of aquatic animals, is beneficial to improvement of aquaculture income, is good in stability and convenient to store, and thus has wide application value.

Description

technical field [0001] The invention belongs to the field of aquaculture, and in particular relates to a compound preparation containing Clostridium butyricum and Pediococcus lactis and its preparation method and application. Background technique [0002] Clostridium butyricum and Pediococcus acidilactici are common Gram-positive bacteria in the environment and in the intestinal tract of animals. Clostridium butyricum can adhere and occupy space in the intestinal tract to inhibit pathogenic bacteria; Lactic acid tablets Probiotics such as pediocin lactate and L-lactic acid can be produced during the reproduction and metabolism of cocci, which play an important role in regulating the balance of intestinal flora. At present, the main problem encountered by these two kinds of microbial agents involved in the preparation is that the production cost of solid products is still high, which limits the frequency of use; and liquid products cannot be used on a large scale due to unsta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K50/80A23K10/18A01K61/59C12R1/145C12R1/01
CPCC12N1/20A23K50/80A23K10/18A01K61/00A23V2400/413Y02A40/818
Inventor 张婷婷王金龙梁晶晶李明霞闫舟梁奕张宗国
Owner QINGDAO SHANGDE BIOTECH
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