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CN-121986869-A - Feed lactobacillus additive and preparation method thereof

CN121986869ACN 121986869 ACN121986869 ACN 121986869ACN-121986869-A

Abstract

The invention relates to the technical field of microbial feed additives, and discloses a feed lactobacillus additive and a preparation method thereof, wherein the preparation method comprises the steps of taking lactobacillus plantarum and/or enterococcus faecalis as strain activation and propagation, and taking agricultural and sideline product waste as a substrate for liquid state deep fermentation to obtain mature fermentation liquor; mixing the mature fermentation broth with a composite protective agent containing double essential oils, chitosan, theabrownin and porous silica, granulating, pre-drying at low temperature to obtain semi-dried granule core, coating with kappa-carrageenan/konjak glucomannan/plant sterol ester composite gel, drying, and packaging to obtain the final product. The double-essential oil, chitosan and theabrownin are combined to form a full-spectrum active antibacterial system, the outer layer of the elastic composite gel realizes double protection of liquid water and water vapor, porous silicon dioxide actively maintains low water activity of particles to prevent lactic acid bacteria from being recovered in advance, and the product has high viable count, strong heat resistance and acid resistance and excellent storage stability.

Inventors

  • WANG PENG
  • HOU YONGWEI
  • LI FUHOU
  • TANG HONGYU
  • YUAN BAO

Assignees

  • 吉林大学

Dates

Publication Date
20260508
Application Date
20260410

Claims (10)

  1. 1. The preparation method of the feed lactobacillus additive is characterized by comprising the following steps of: inoculating lactobacillus into a liquid culture medium for activation culture to obtain first-stage seed liquid; Preparing a liquid fermentation medium by taking agricultural and sideline product waste as a carbon source and a nitrogen source, and sterilizing; Step three, inoculating the first-stage seed liquid into a liquid fermentation medium for closed fermentation until the viable count is not lower than CFU/mL to obtain mature fermentation liquor; step four, mixing porous starch, skimmed milk powder, trehalose, sodium glutamate, natural plant essential oil, food-grade chitosan, black tea theabrownin and food-grade porous silica to obtain a composite protective agent; mixing and granulating the mature fermentation liquid and the composite protective agent according to the mass ratio of 1:1.5-2.5 to obtain wet granules; step six, drying the wet particles at 30-40 ℃ until the moisture content is 8% -12%, so as to obtain a semi-dry particle core; And seventhly, preparing coating liquid from kappa-carrageenan, konjak glucomannan and plant sterol ester, spraying the coating liquid onto the surface of the semi-dried granule core, cooling, solidifying and drying until the moisture content is lower than 8%, thus obtaining the feed lactobacillus additive.
  2. 2. The preparation method of the lactobacillus plantarum/enterococcus faecalis, which is characterized in that the lactobacillus in the first step is at least one of lactobacillus plantarum and enterococcus faecalis, the liquid culture medium is MRS liquid culture medium, and the activating culture condition is 35-38 ℃ for static culture for 18-24 hours.
  3. 3. The method according to claim 1, wherein in the second step, the agricultural and sideline product waste is one or more of horseshoe starch processing wastewater, tapioca dreg water extract or bran water extract; Leaching the cassava residue water leaching solution for 1-2 hours at the temperature of 60-80 ℃ according to the solid-to-liquid ratio of 1:10-1:15, and filtering to obtain a supernatant; leaching the bran water extract for 1-2 hours at the temperature of 60-80 ℃ according to the solid-to-liquid ratio of 1:8-1:12, and filtering to obtain a supernatant; the fermentation medium was sterilized at 115 ℃ for 30 minutes.
  4. 4. The preparation method of the composite protective agent according to claim 1 is characterized in that the composite protective agent in the fourth step comprises, by weight, 25-40 parts of porous starch, 20-30 parts of skimmed milk powder, 10-20 parts of trehalose, 5-10 parts of sodium glutamate, 5-10 parts of natural plant essential oil, 5-10 parts of food-grade chitosan, 5-8 parts of black tea theabrownin and 3-8 parts of food-grade porous silica.
  5. 5. The preparation method of claim 4, wherein in the fourth step, the natural plant essential oil is a mixture of thymol/carvacrol essential oil and clove essential oil, the mass ratio of the thymol/carvacrol essential oil to the mixture is 1-2:1, the total content of thymol and carvacrol in the thymol/carvacrol essential oil is not less than 80%, the thymol content accounts for 50% -70% of the total content, and the eugenol content in the clove essential oil is not less than 75%.
  6. 6. The preparation method of claim 4, wherein in the step four, the deacetylation degree of the food-grade chitosan is not lower than 85%, the viscosity average molecular weight is 10-20 Da, the food-grade porous silica is amorphous silica, the specific surface area is not lower than 150m 2/g.
  7. 7. The preparation method of claim 4, wherein the natural plant essential oil in the fourth step is introduced by adopting a pre-adsorption mode, namely, after thymol/carvacrol essential oil is mixed with clove essential oil, the mixture is added into 40% -60% of the total amount of the porous starch used in the fourth step, and stirring is carried out for 15-30 minutes until the appearance of the mixture is free-flowing dry powder, so that porous starch containing essential oil is obtained, and the porous starch is uniformly mixed with the other components in the fourth step.
  8. 8. The preparation method of the coating liquid according to claim 1, wherein the coating liquid in the seventh step comprises 3-5 parts by weight of kappa-carrageenan, 1-2 parts by weight of konjak glucomannan, 1-2 parts by weight of plant sterol ester and 0.3-0.8 part by weight of feed-grade potassium chloride, and the total concentration of kappa-carrageenan and konjak glucomannan in the coating liquid is 3% -6%.
  9. 9. The preparation method of the coating liquid according to claim 8, wherein the preparation method of the coating liquid in the seventh step is characterized in that kappa-carrageenan, konjak glucomannan and potassium chloride are added into water and heated to 70-80 ℃ to be completely dissolved, a polymer solution is obtained, plant sterol ester is independently preheated to 70-80 ℃ to be completely melted and then added into the polymer solution, stirring is carried out for 10-15 minutes at 70-80 ℃ at 3000-5000 r/min, the obtained coating liquid is kept at 60-65 ℃ for standby, a fluidized bed coating machine is used for spraying, the coating amount is 5% -12% of the total mass of particles, and the composite gel is solidified after coating and cooling to below 35 ℃.
  10. 10. A feed lactic acid bacteria additive characterized by being produced by the production method according to any one of claims 1 to 9, the viable count of the feed lactic acid bacteria additive being not less than CFU/g, survival rate of viable bacteria not lower than 70% after 5min treatment at 85 ℃, survival rate of viable bacteria not lower than 80% after 2h treatment in artificial gastric juice at pH 2.5, and retention rate of viable bacteria not lower than 90% after sealed preservation at 4 ℃ for 12 months.

Description

Feed lactobacillus additive and preparation method thereof Technical Field The invention relates to the technical field of microbial feed additives, in particular to a feed lactobacillus additive and a preparation method thereof. Background With the development of antibiotic bans for global feed, microbial probiotic additives for feed have become an important antibiotic alternative for animal husbandry. Lactic acid bacteria are widely adopted by virtue of the effects of regulating intestinal microecology, inhibiting pathogenic bacteria and improving body immunity, and are one of the most potential microbial feed additives at present. The lactobacillus additive is generally prepared into solid granular products by microencapsulation or carrier adsorption after high-concentration viable bacteria are obtained by liquid fermentation, so as to protect the viable bacteria from survival during storage and feeding. The commercial products at present usually take inert physical protective components such as skimmed milk powder, starch and the like as protective agents, water-soluble materials as outer layers for coating, and special commercial culture mediums such as MRS and the like for liquid fermentation production. However, the existing feed lactobacillus additive still has obvious defects in practical production and application, the existing protective agent formula does not contain active antibacterial components, the product is easy to be polluted by exogenous mixed bacteria in the storage period, the viable count is continuously reduced, the existing water-soluble outer layer is rapidly dissolved and shed in feeding environments such as farm trough ponding, high humidity climate, alternate dry and wet and the like or microcracks are generated due to repeated expansion and shrinkage, so that the lactobacillus is greatly attenuated before being ingested by animals, and in addition, the traditional liquid fermentation depends on special commercial culture media such as MRS and the like, the production cost is high, and the large-scale industrial production is not facilitated. Disclosure of Invention The invention provides a feed lactobacillus additive and a preparation method thereof, which solve the technical problems that the existing feed lactobacillus additive in the related art lacks an active antibacterial function, a protective layer fails in a wet farm environment and the production cost is high. The invention provides a preparation method of a feed lactobacillus additive, which comprises the following steps: inoculating lactobacillus into a liquid culture medium for activation culture to obtain first-stage seed liquid; Preparing a liquid fermentation medium by taking agricultural and sideline product waste as a carbon source and a nitrogen source, and sterilizing; Step three, inoculating the first-stage seed liquid into a liquid fermentation medium for closed fermentation until the viable count is not lower than CFU/mL to obtain mature fermentation liquor; step four, mixing porous starch, skimmed milk powder, trehalose, sodium glutamate, natural plant essential oil, food-grade chitosan, black tea theabrownin and food-grade porous silica to obtain a composite protective agent; Mixing and granulating the mature fermentation liquid and a composite protective agent according to the mass ratio of 1 (1.5-2.5) to obtain wet granules; step six, drying the wet particles at 30-40 ℃ until the moisture content is 8% -12%, so as to obtain a semi-dry particle core; And seventhly, preparing coating liquid from kappa-carrageenan, konjak glucomannan and plant sterol ester, spraying the coating liquid onto the surface of the semi-dried granule core, cooling, solidifying and drying until the moisture content is lower than 8%, thus obtaining the feed lactobacillus additive. Preferably, the lactobacillus in the first step is at least one of lactobacillus plantarum (Lactobacillus plantarum) and enterococcus faecalis (Enterococcus faecalis), the liquid culture medium is MRS liquid culture medium, and the activating culture condition is that standing culture is carried out for 18-24 hours at 35-38 ℃. The agricultural and sideline product waste in the second step is one or more of water chestnut starch processing wastewater, cassava dreg water extract or bran water extract, wherein the cassava dreg water extract is leached for 1-2 hours at a solid-to-liquid ratio of 1:10-1:15 (w/v), supernatant is filtered, the bran water extract is leached for 1-2 hours at a solid-to-liquid ratio of 1:8-1:12 (w/v), supernatant is filtered, and the fermentation medium is sterilized for 30 minutes at a temperature of 115 ℃. Preferably, the composite protective agent in the fourth step comprises, by weight, 25-40 parts of porous starch (oil absorption not lower than 80%, w/w), 20-30 parts of skim milk powder, 10-20 parts of trehalose, 5-10 parts of sodium glutamate, 5-10 parts of natural plant essential oil, 5-10 parts of food-grade chitosan, 5-