阅读说明：本技术 一种仔猪肠道健康调控活性肽饲料添加剂及应用 (Piglet intestinal health regulation and control active peptide feed additive and application thereof ) 是由 徐求文 金妙仁 楼伟杰 于晓莹 洪奇华 于 2019-11-15 设计创作，主要内容包括：本发明公开了一种仔猪肠道健康调控活性肽饲料添加剂及应用,所述活性肽饲料添加剂按如下方法制备：将谷朊粉和蚕蛹粉混合并加水搅拌均匀,再加入木瓜蛋白酶和胰蛋白酶,调节pH至7.0-8.0,在50-55℃条件下酶解30-60min,获得酶解物；将枯草芽孢杆菌、酿酒酵母接种至由酶解物配制的固态发酵培养基中,33～37℃共混发酵培养30～45小时,发酵混合物干燥,获得所述的活性肽饲料添加剂；应用本发明的仔猪肠道健康调控活性肽,能防止仔猪断奶后1～7天肠黏膜绒毛过度萎缩和隐窝的加深,可显著提高断奶后8～14天肠黏膜绒毛高度。(The invention discloses a piglet intestinal health regulation active peptide feed additive and application thereof, wherein the active peptide feed additive is prepared by the following method: mixing wheat gluten and pupa Bombycis powder, adding water, stirring, adding papain and trypsin, adjusting pH to 7.0-8.0, and performing enzymolysis at 50-55 deg.C for 30-60min to obtain zymolyte; inoculating bacillus subtilis and saccharomyces cerevisiae into a solid fermentation culture medium prepared from zymolyte, performing blending fermentation culture at 33-37 ℃ for 30-45 hours, and drying a fermentation mixture to obtain the active peptide feed additive; by applying the piglet intestinal health regulation active peptide, excessive shrinkage of intestinal mucosa villi and deepening of crypt after piglets weaning for 1-7 days can be prevented, and intestinal mucosa villi height after weaning for 8-14 days can be remarkably improved.)

1. The active peptide feed additive for regulating and controlling the intestinal health of the piglets is characterized by being prepared by the following method: (1) mixing wheat gluten and pupa Bombycis powder, adding water, stirring, adding papain and trypsin, adjusting pH to 7.0-8.0, and performing enzymolysis at 50-55 deg.C for 30-60min to obtain zymolyte; (2) inoculating bacillus subtilis and saccharomyces cerevisiae into a solid fermentation culture medium prepared by the zymolyte in the step (1), performing blending fermentation culture at 33-37 ℃ for 30-45 hours, and drying a fermentation mixture to obtain the active peptide feed additive; the solid state fermentation culture medium comprises the following components in percentage by weight: 50-65% of an enzymatic hydrolysate KH₂PO₄0.15-0.25% of wheat bran 29.35-40.25%, 2-4.5% of soybean meal, 2-3% of molasses and 1.5-2% of urea.

2. The piglet intestinal health regulation active peptide feed additive as claimed in claim 1, wherein the mass ratio of the silkworm chrysalis meal to the wheat gluten in the step (1) is 1: 5-10; the adding amount of the water is 3-5 times of the total weight of the vital gluten and the silkworm chrysalis meal.

3. The active peptide feed additive for regulating intestinal health of piglets as claimed in claim 1, wherein in step (1), the amount of papain added is 1-1.5% of the total weight of the gluten powder and the silkworm chrysalis powder, and the amount of trypsin added is 0.5-1% of the total weight of the gluten powder and the silkworm chrysalis powder.

4. The piglet intestinal health control active peptide feed additive as claimed in claim 1, wherein the enzymolysis in step (1) is carried out under the assistance of 400W and 25kHz ultrasound.

5. The piglet intestine health supplement of claim 1The controlled-activity peptide feed additive is characterized in that in the step (2), the Bacillus subtilis is Bacillus subtilis ACCC01185, and the inoculation amount of the Bacillus subtilis is 2.5-50 multiplied by 10⁷cfu/g。

6. The piglet intestinal health control active peptide feed additive of claim 1, wherein the Saccharomyces cerevisiae in the step (2) is (Saccharomyces cerevisiae) ACCC20042, and the inoculation amount of the Saccharomyces cerevisiae is 1.5-3.6 x 10⁷cfu/g。

7. The piglet intestinal health control bioactive peptide feed additive as claimed in claim 1, wherein the bacillus subtilis is inoculated in step (2) in the form of seed solution, and the seed solution is prepared by the following method: inoculating bacillus subtilis to a slant culture medium, and culturing at 37 ℃ for 24h to obtain slant strains; inoculating a circulant lawn from the inclined plane to a 300ml triangular flask seed filled with 50ml of seed culture medium, placing the triangular flask seed in a 35-37 ℃ temperature-controlled shaking table for culturing at the rotating speed of 220r/min for 24-26h to obtain the bacillus subtilis seed liquid; the slant culture medium comprises: 10g/L of peptone, 3g/L, NaC 15 g/15 g/L of beef extract, 20g/L of agar, distilled water as solvent and 7.0 of pH value; the seed culture medium comprises the following components: 2g/L of glucose, 15 g/L of NaC 15 g/L, 5g/L of yeast extract, 0g/L of peptone L, distilled water as a solvent and pH 7.0.

8. The piglet intestinal health control active peptide feed additive as claimed in claim 1, wherein the saccharomyces cerevisiae in step (2) is added in the form of yeast seed solution, and the seed solution is prepared by the following method: inoculating Saccharomyces cerevisiae into a triangular flask filled with 100ml maltose liquid, and culturing at 30-32 deg.C for 12 hr; transferring to a triangular flask filled with 600ml maltose liquid, and culturing at 30-32 deg.C for 12 h; then transferring the yeast into a Kaschin jar filled with 12kg of sugar water, culturing for 12h at 30-32 ℃, finally transferring the yeast into a yeast jar filled with a culture medium, and culturing for 18-20h at 30-32 ℃ to obtain yeast seed liquid, wherein the culture medium comprises 300kg of clear water, 30kg of flour and 10kg of bran in each jar; the sugar degrees of the maltose liquid and the sugar liquid are both 7-8 degrees Be.

9. The use of the piglet intestinal health control bioactive peptide feed additive of claim 1 in preparing piglet feed.

10. The use according to claim 9, characterized in that the feed is a piglet gut health control feed.

(I) technical field

The invention relates to a piglet intestinal health regulation active peptide feed additive and a preparation method and application thereof.

(II) background of the invention

The early weaning can improve the reproductive capacity of the sow, increase the annual number born of the sow, reduce the disease transmission from the mother to the piglet, improve the production performance and carcass quality of the growing pig and improve the utilization rate of equipment such as a delivery room and the like. However, an important problem brought by early weaning is how to improve the growth performance of weaned piglets, the properties of daily ration of the weaned piglets are changed greatly, the digestive tract of the piglets is not developed completely, the digestive enzyme activity is insufficient, the immune function is not developed to be mature, and the weaning is not only related to the physiological change, the psychological, environmental and nutritional conditions, but also changed, so that the piglet early weaning syndrome is often expressed by so-called piglet early weaning syndrome of poor appetite, digestive dysfunction, diarrhea, growth retardation, low feed utilization rate and the like.

A great deal of research shows that Glutamine (GLN) is a fuel of intestinal cells, is a precursor of purine and pyrimidine nucleotides necessary for the differentiation of synthetic cells, is an important energy substrate of mesenteric lymph and epithelial lymphocytes, is an essential component for maintaining lymph tissues related to the intestinal tract, is an essential substance for synthesizing immunoglobulin secreted by the intestinal tract, and plays an important role in maintaining the normal function of the intestinal tract and improving the immunity of the intestinal tract (James, 1998). Furthermore, GLN is the most abundant amino acid in pig milk, and the decrease in GLN supply after weaning is the most fundamental cause of early weaning syndrome in piglets (Alverdy, 1990). The exogenous small molecule active peptide can reduce the absorption competition of free amino acid, promote the secretion of hormone, promote the absorption and utilization of mineral elements, improve the immunity of organisms and improve the production performance. Therefore, the feed additive rich in GLN and active small peptide has a remarkable effect of relieving early weaning syndrome of piglets.

Disclosure of the invention

The invention aims to provide a piglet intestinal health regulation active peptide feed additive and a preparation method and application thereof.

The technical scheme adopted by the invention is as follows:

the invention provides a piglet intestinal health regulation active peptide feed additive, which is prepared by the following steps: (1) mixing wheat gluten and pupa Bombycis powder, adding water, stirring, adding papain and trypsin, adjusting pH to 7.0-8.0, and performing enzymolysis at 50-55 deg.C for 30-60min to obtain zymolyte; (2) inoculating Bacillus subtilis and Saccharomyces cerevisiae into a solid fermentation culture medium prepared by the zymolyte in the step (1), performing blending fermentation culture at 33-37 ℃ for 30-45 hours, and drying a fermentation mixture to obtain the active peptide feed additive; the solid state fermentation culture medium comprises the following components in percentage by weight: 50-65% of an enzymatic hydrolysate KH₂PO₄0.15-0.25% of wheat bran 29.35-40.25%, 2-4.5% of soybean meal, 2-3% of molasses and 1.5-2% of urea.

In the step (1), the mass ratio of the silkworm chrysalis meal to the wheat gluten is 1: 5-10; the adding amount of the water is 3-5 times of the total weight of the vital gluten and the silkworm chrysalis meal; the adding amount of the papain is 1-1.5% of the total weight of the vital gluten and the silkworm chrysalis meal, and the adding amount of the trypsin is 0.5-1% of the total weight of the vital gluten and the silkworm chrysalis meal. The enzyme activity of the papain is 8-10 ten thousand U/g, and the enzyme activity of the trypsin is 3000-4000U/g.

And (2) carrying out enzymolysis under the assistance of 400W and 25kHz ultrasonic waves.

In the step (2), the Bacillus subtilis is Bacillus subtilis ACCC01185 and is purchased from China agricultural microbial strain preservation management center; the amount of inoculation may be unlimited, and may be selected from conventional amounts. Preferably, the inoculation amount of the bacillus subtilis is 2.5-50 multiplied by 10 based on the dry weight of the solid fermentation medium⁷cfu/g。

The Saccharomyces cerevisiae in the step (2) is (Saccharomyces cerevisiae) ACCC20042, which is purchased from China agricultural microbial strain preservation management center, and the inoculation amount can be unlimited, and can be selected generally. Preferably, the inoculation amount of the saccharomyces cerevisiae is 1.5-3.6 multiplied by 10 based on the dry weight of the solid fermentation medium⁷cfu/g。

Inoculating the bacillus subtilis in the step (2) in a seed liquid form, wherein the seed liquid is prepared by the following method: inoculating bacillus subtilis to a slant culture medium, and culturing at 37 ℃ for 24h to obtain slant strains; inoculating a circulant lawn from the inclined plane to a 300ml triangular flask seed filled with 50ml of seed culture medium, placing the triangular flask seed in a 35-37 ℃ temperature-controlled shaking table for culturing at the rotating speed of 220r/min for 24-26h to obtain the bacillus subtilis seed liquid; the slant culture medium comprises: 10g/L of peptone, 3g/L, NaC 15 g/15 g/L of beef extract, 20g/L of agar, distilled water as solvent and 7.0 of pH value; the seed culture medium comprises the following components: 2g/L of glucose, 15 g/L of NaC 15 g/L, 5g/L of yeast extract, 0g/L of peptone L, distilled water as a solvent and pH7.0.

Adding the saccharomyces cerevisiae in the step (2) in a form of yeast seed liquid, wherein the seed liquid is prepared by the following method: the method comprises the steps of performing heat preservation culture at 30-32 ℃ for 12h in a saccharomyces cerevisiae protobacteria test tube-small triangular flask seeds (maltose liquid 100ml, the sugar degree of 7-8 degrees Be), transferring to a large triangular flask (maltose liquid 600ml, the sugar degree of 7-8 degrees Be), performing heat preservation culture at 30-32 ℃ for 12h, transferring to a Ka tank (sugar liquid 12kg, the sugar degree of 7-8 degrees Be amplification culture) for 12h at 30-32 ℃, performing expansion culture on the Ka tank yeast seeds in a yeast tank (culture medium is 300kg of clear water, 30kg of wheat flour and 10kg of bran in each tank), and performing culture at 30-32 ℃ for 18-20h to obtain yeast seed liquid.

And (2) drying until the water content mass concentration is not more than 10%.

The invention also provides an application of the piglet intestinal health regulation active peptide feed additive in piglet feed, and the feed is feed for regulating and controlling piglet intestinal health.

Compared with the prior art, the invention has the following beneficial effects:

starting from the characteristics of digestive physiological development of piglets, the invention applies nutrition and immunity regulation and control means to carry out enzymolysis and fermentation on the combination of the vital gluten and the silkworm chrysalis powder, and successfully develops the piglet intestinal health regulation and control active peptide feed additive. By applying the piglet intestinal health regulation active peptide, excessive shrinkage of intestinal mucosa villi and deepening of crypt after piglets weaning for 1-7 days can be prevented, and intestinal mucosa villi height after weaning for 8-14 days can be remarkably improved. The piglet intestinal health regulation active peptide can obviously improve the average daily gain of weaned piglets and reduce the piglet weaning diarrhea rate, and has obvious economic and social benefits and wide market prospect.

(IV) detailed description of the preferred embodiments

The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto: