A repair agent for reducing diarrhea in weaned piglets

Abstract

The present invention provides a repair agent for reducing diarrhea in weaned piglets, belonging to the technical field of animal medicine. The present invention uses the shrimp meat of Acanthomysis longirostris and Acanthomysis gracilipes as raw materials to prepare a composite polypeptide. Moreover, through experiments, it is found that the composite polypeptide prepared by the present invention can significantly promote the proliferation and migration of porcine intestinal epithelial cells, so that the composite polypeptide prepared by the present invention can be used to repair porcine intestinal damage, reduce diarrhea in piglets, and improve the survival rate of piglets.

Description

[0001] This case is a divisional application. The name of the invention of the original application is: A repair agent for pig intestinal repair. The application date of the original application is: 2020-10-15, and the application number of the original application is: CN202011100442.5. Technical Field

[0002] The invention belongs to the technical field of animal medicine, and in particular relates to a repair agent for reducing diarrhea in weaned piglets. Background Art

[0003] With the continuous development of our country's economic strength, people's living standards have been continuously improved, and the demand for pork has also been continuously increased. my country is the country with the largest pork production and consumption in the world. The healthy development of the pig industry plays an important role in the macro-control of the national economy. Among them, the survival rate of piglets is an important factor affecting the economic benefits of pig farms and pork prices. In my country's pork farming, diarrhea in weaned piglets is the main reason affecting the survival rate of piglets. The incidence of diarrhea in weaned piglets in my country is usually about 20%-30%, and the mortality rate of weaned piglets is 10-15%.

[0004] Diarrhea in piglets is mostly caused by damage to the intestinal mucosal epithelial tissue of piglets. There are two ways to repair the intestinal epithelium: one is that the cells adjacent to the damaged surface migrate to cover the damaged area; the second is to replace the damaged cells through cell proliferation. Therefore, finding substances that can promote the proliferation and migration of epithelial cells in the pig intestine can effectively repair intestinal epithelial damage, thereby effectively reducing diarrhea in piglets and improving the survival rate of piglets. Summary of the invention

[0005] The purpose of the present invention is to provide a repairing agent for repairing the intestinal tract of pigs.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] The invention provides use of a composite polypeptide in preparing a pig intestinal repair agent. The raw materials for preparing the composite polypeptide are long-fronted spiny mysid shrimp meat and small red mysid shrimp meat.

[0008] Preferably, the preparation method of the composite polypeptide is as follows:

[0009] (1) Crush and mix the long-fronted spiny shrimp meat and the small red mysid shrimp meat in a ratio of 1:1, add 15 times the amount of water to make a homogenate, and sterilize for later use;

[0010] (2) Add 3 wt% Bacillus subtilis, 35 °C, pH 6, 30 r / min, and ferment for 48 h;

[0011] (3) After fermentation, sterilize with high temperature and high pressure for 20 minutes;

[0012] (4) After cooling to room temperature, centrifuge the sterilized fermentation broth at 3500 rpm for 20 min;

[0013] (5) Take the supernatant and filter it using a 1 μm ultrafiltration membrane to obtain a polypeptide solution;

[0014] (6) Freeze-drying to obtain the composite peptides of long-fronted spiny shrimp and small red shrimp.

[0015] In addition, the present invention provides the use of the composite polypeptide in the preparation of a promoter for promoting the proliferation of porcine small intestinal epithelial cells, and the preparation method of the composite polypeptide is as follows:

[0016] (1) Crush and mix the long-fronted spiny shrimp meat and the small red mysid shrimp meat in a ratio of 1:1, add 15 times the amount of water to make a homogenate, and sterilize for later use;

[0017] (2) Add 3 wt% Bacillus subtilis, 35 °C, pH 6, 30 r / min, and ferment for 48 h;

[0018] (3) After fermentation, sterilize with high temperature and high pressure for 20 minutes;

[0019] (4) After cooling to room temperature, centrifuge the sterilized fermentation broth at 3500 rpm for 20 min;

[0020] (5) Take the supernatant and filter it using a 1 μm ultrafiltration membrane to obtain a polypeptide solution;

[0021] (6) Freeze-drying to obtain the composite peptides of long-fronted spiny shrimp and small red shrimp.

[0022] In addition, the present invention provides the use of the composite polypeptide in the preparation of an agent for promoting the migration of porcine small intestinal epithelial cells, and the preparation method of the composite polypeptide is as follows:

[0023] (1) Crush and mix the long-fronted spiny shrimp meat and the small red mysid shrimp meat in a ratio of 1:1, add 15 times the amount of water to make a homogenate, and sterilize for later use;

[0024] (2) Add 3 wt% Bacillus subtilis, 35 °C, pH 6, 30 r / min, and ferment for 48 h;

[0025] (3) After fermentation, sterilize with high temperature and high pressure for 20 minutes;

[0026] (4) After cooling to room temperature, centrifuge the sterilized fermentation broth at 3500 rpm for 20 min;

[0027] (5) Take the supernatant and filter it using a 1 μm ultrafiltration membrane to obtain a polypeptide solution;

[0028] (6) Freeze-drying to obtain the composite peptides of long-fronted spiny shrimp and small red shrimp.

[0029] In addition, the present invention provides a repairing agent for repairing the intestinal tract of pigs, wherein the repairing agent is a composite polypeptide, and the raw materials for preparing the composite polypeptide are the meat of long-fronted spiny shrimp and the meat of small red mysid shrimp.

[0030] Preferably, the preparation method of the composite polypeptide is as follows:

[0031] (1) Crush and mix the long-fronted spiny shrimp meat and the small red mysid shrimp meat in a ratio of 1:1, add 15 times the amount of water to make a homogenate, sterilize and set aside;

[0032] (2) Add 3 wt% Bacillus subtilis, 35 °C, pH 6, 30 r / min, and ferment for 48 h;

[0033] (3) After fermentation, sterilize with high temperature and high pressure for 20 minutes;

[0034] (4) After cooling to room temperature, centrifuge the sterilized fermentation broth at 3500 rpm for 20 min;

[0035] (5) Take the supernatant and filter it using a 1 μm ultrafiltration membrane to obtain a polypeptide solution;

[0036] (6) Freeze-drying to obtain the composite peptides of long-fronted spiny shrimp and small red shrimp.

[0037] In addition, the present invention provides a promoter for promoting the proliferation of pig small intestinal epithelial cells, wherein the promoter is a composite polypeptide, and the raw materials for preparing the composite polypeptide are long-fronted spiny mysid shrimp meat and small red mysid shrimp meat;

[0038] The preparation method of the composite polypeptide is as follows:

[0039] (1) Crush and mix the long-fronted spiny shrimp meat and the small red mysid shrimp meat in a ratio of 1:1, add 15 times the amount of water to make a homogenate, sterilize and set aside;

[0040] (2) Add 3 wt% Bacillus subtilis, 35 °C, pH 6, 30 r / min, and ferment for 48 h;

[0041] (3) After fermentation, sterilize with high temperature and high pressure for 20 minutes;

[0042] (4) After cooling to room temperature, centrifuge the sterilized fermentation broth at 3500 rpm for 20 min;

[0043] (5) Take the supernatant and filter it using a 1 μm ultrafiltration membrane to obtain a polypeptide solution;

[0044] (6) Freeze-drying to obtain the composite peptides of long-fronted spiny shrimp and small red shrimp.

[0045] In addition, the present invention provides an accelerator for promoting the migration of pig small intestinal epithelial cells, wherein the accelerator is a composite polypeptide, and the raw materials for preparing the composite polypeptide are long-fronted spiny mysid shrimp meat and small red mysid shrimp meat;

[0046] The preparation method of the composite polypeptide is as follows:

[0047] (1) Crush and mix the long-fronted spiny shrimp meat and the small red mysid shrimp meat in a ratio of 1:1, add 15 times the amount of water to make a homogenate, sterilize and set aside;

[0048] (2) Add 3 wt% Bacillus subtilis, 35 °C, pH 6, 30 r / min, and ferment for 48 h;

[0049] (3) After fermentation, sterilize with high temperature and high pressure for 20 minutes;

[0050] (4) After cooling to room temperature, centrifuge the sterilized fermentation broth at 3500 rpm for 20 min;

[0051] (5) Take the supernatant and filter it using a 1 μm ultrafiltration membrane to obtain a polypeptide solution;

[0052] (6) Freeze-drying to obtain the composite peptides of long-fronted spiny shrimp and small red shrimp.

[0053] The beneficial effects of the present invention are:

[0054] The present invention uses long-fronted spiny shrimp and small red shrimp as raw materials to prepare a composite polypeptide for pig intestinal repair. The composite polypeptide can effectively promote the proliferation of pig small intestinal epithelial cells and promote the migration of pig small intestinal epithelial cells, thereby effectively repairing the pig intestine. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 Effects of compound peptides on the proliferation of porcine intestinal epithelial cells IPEC-J2

[0056] Figure 2 Effects of compound peptides on migration of porcine intestinal epithelial cells IPEC-J2 DETAILED DESCRIPTION

[0057] Example 1

[0058] (1) Crush and mix 50 g of long-fronted spiny shrimp meat and 50 g of small red shrimp meat, add 1500 ml of water to make a homogenate, and sterilize for later use;

[0059] (2) Add 60 g of Bacillus subtilis, 35°C, pH 6, 30 r / min, and ferment for 48 hours;

[0060] (3) After fermentation, sterilize at high temperature and high pressure for 20 min;

[0061] (4) After cooling to room temperature, centrifuge the sterilized fermentation broth at 3500 rpm for 20 min;

[0062] (5) Take the supernatant and filter it using a 1-μm ultrafiltration membrane to obtain a polypeptide solution;

[0063] (6) Freeze-dry to obtain the composite polypeptide of *Acanthomysis longirostris* and *Mysis graciloides*.

[0064] Example 2

[0065] Application of the composite polypeptide of *Acanthomysis longirostris* and *Mysis graciloides* in promoting the proliferation of porcine intestinal epithelial cells.

[0066] (1) Inoculate 1×10 4 IPEC-J2 cells into a 96-well plate. After overnight culture, add a complex protease for treatment (0 μg / ml, 10 μg / ml, 25 μg / ml, 50 μg / ml);

[0067] (2) After treatment for 48 h, use a CCK-8 reagent to detect the OD value. The results are shown in Table 1 and Figure 1 as follows.

[0068] Table 1 OD values detected by CCK-8 (statistical differences are as Figure 1 shown)

[0069]

[0070] It can be seen from Figure 1 and Table 1 that the composite polypeptide prepared by the present invention can significantly promote the proliferation of porcine small intestinal epithelial cells IPEC-J2, and the difference is statistically significant.

[0071] Example 3

[0072] (1) Inoculate IPEC-J2 cells into a 6-well plate. When the cell density reaches 85%, gently slide a 200-μl pipette tip across the adherent cell layer in the vertical direction of the culture well, and use PBS to remove the detached cells;

[0073] (2) Add media containing 0 μg / mL and 50 μg / mL, and take pictures using an inverted microscope at 0 h and 48 h respectively. The experimental results are as Figure 2 shown.

[0074] It can be seen from Figure 2 that the composite polypeptide can significantly promote the migration ability of porcine small intestinal epithelial cells IPEC-J2.

[0075] In summary, on the one hand, the composite polypeptide prepared by the present invention can replace damaged cells by promoting the proliferation of porcine small intestinal epithelial cells, and on the other hand, it can cover the damaged area by promoting the migration of porcine small intestinal epithelial cells. Therefore, it can be used to prepare a repair agent for repairing porcine intestinal damage to reduce diarrhea in weaned piglets and improve the survival rate of weaned piglets.

Claims

1. Use of a composite polypeptide in the preparation of a repair agent for reducing diarrhea in weaned piglets, characterized in that, The preparation method of the composite polypeptide is as follows: (1) Crush and mix the long-headed spiny shrimp meat and the small red mysid shrimp meat in a ratio of 1:1, add 15 times the amount of water to make a homogenate, and sterilize for later use; (2) adding 3 wt % Bacillus subtilis, 35 ° C, pH 6, 30 r / min, and fermenting for 48 h; (3) After fermentation, sterilize under high temperature and high pressure for 20 min; (4) After cooling to room temperature, the sterilized fermentation broth was centrifuged at 3500 rpm for 20 min; (5) taking the supernatant and filtering it using a 1 μm ultrafiltration membrane to obtain a polypeptide solution; (6) freeze-drying to obtain the composite polypeptide of the long-fronted spiny shrimp and the small red shrimp.

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