CN112168950A - Repairing agent for repairing pig intestinal tract - Google Patents
Repairing agent for repairing pig intestinal tract Download PDFInfo
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Abstract
The invention provides a repairing agent for repairing pig intestinal tracts, and belongs to the technical field of animal medicine. According to the invention, the compound polypeptide is prepared by using the prawns meat of the oplophorus macrolepis and the small red prawns as raw materials, and a CCK-8 experiment and a cell scratch experiment prove that the compound polypeptide prepared by the invention can obviously promote the proliferation and migration of epithelial cells of small intestines of pigs, so that the compound polypeptide prepared by the invention can be used for preparing a pig intestinal tract repairing agent to repair the pig intestinal tract injury, reduce the diarrhea of piglets and improve the survival rate of the piglets.
Description
Technical Field
The invention belongs to the technical field of animal medicine, and particularly relates to a repairing agent for repairing porcine intestinal tracts.
Background
Along with the continuous development of the economic strength of our country, the living standard of people is continuously improved, and the requirement of pork is also continuously increased. China is the country with the largest pork yield and consumption in the world, and the healthy development of the pig raising industry plays an important role in the macroscopic regulation and control of national economy. The survival rate of piglets is an important factor influencing the economic benefit of a pig farm and the price of pork. In pork cultivation in China, diarrhea of weaned piglets is the most main reason for influencing the survival rate of piglets. The diarrhea incidence of the weaned pigs in China is generally about 20% -30%, and the fatality rate of the weaned pigs is 10-15%.
The diarrhea of piglets is caused by the damage of intestinal mucosal epithelial tissues of the piglets. There are two ways of repair of intestinal epithelium: one way is that cells adjacent to the damaged surface migrate to cover the damaged area; the second way is to replace damaged cells by cell proliferation. Therefore, substances capable of storing the proliferation of the epithelial cells in the intestinal tract of the pig and the migration of the epithelial cells are searched, and the intestinal epithelial injury can be effectively repaired, so that the diarrhea of piglets is effectively reduced, and the survival rate of the piglets is improved.
Disclosure of Invention
The invention aims to provide a repairing agent for repairing pig intestinal tracts.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides application of composite polypeptide in preparation of a pig intestinal tract repairing agent, wherein the composite polypeptide is prepared from raw materials of prawns of longforehead opyron and prawns of small red prawns.
Preferably, the preparation method of the composite polypeptide is as follows:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
In addition, the invention provides an application of the compound polypeptide in preparing the promoter for promoting the proliferation of the porcine small intestinal epithelial cells, and the preparation method of the compound polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
In addition, the invention provides an application of the compound polypeptide in preparing the promoter for promoting the migration of the epithelial cells of the small intestine of the pig, and the preparation method of the compound polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
In addition, the invention provides a repairing agent for repairing pig intestinal tracts, wherein the repairing agent is a composite polypeptide, and the preparation raw materials of the composite polypeptide are prawns of longforehead acanthosis and prawns of small red prawns.
Preferably, the preparation method of the composite polypeptide is as follows:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
In addition, the invention provides an accelerant for promoting the proliferation of the porcine small intestinal epithelial cells, the accelerant is a composite polypeptide, and the preparation raw materials of the composite polypeptide are prawns of red-skin, and the accelerant is prepared from prawns of pra;
the preparation method of the composite polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
In addition, the invention provides an accelerant for promoting the migration of the epithelial cells of the small intestine of the pig, the accelerant is a composite polypeptide, and the preparation raw materials of the composite polypeptide are the shrimp meat of the prawns with the long forehead and the shrimp meat of the small red prawns;
the preparation method of the composite polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
The invention has the beneficial effects that:
the compound polypeptide for repairing the pig intestinal tract is prepared by using the long-nosed mysidacea and the small red mysidacea as raw materials, can effectively promote the proliferation of the epithelial cells of the pig small intestine and the migration of the epithelial cells of the pig small intestine, and thus can effectively repair the pig intestinal tract.
Drawings
FIG. 1 Effect of Complex Polypeptides on cell proliferation of porcine Small intestinal epithelial cell IPEC-J2
FIG. 2 Effect of the composite polypeptide on cell migration of porcine small intestine epithelial cells IPEC-J2.
Detailed Description
Example 1
(1) Pulverizing and mixing 50g of long-forehead oplophorus macrolepis shrimp meat and 50g of small-red oplophorus macrolepis shrimp meat, adding 1500ml of water to prepare homogenate, and sterilizing for later use;
(2) adding 60g of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
Example 2
Application of composite polypeptide of prawns with oplophorax elatus and small red prawns in promoting proliferation of epithelial cells in intestinal tracts of pigs.
(1) Will be 1 × 104The IPEC-J2 cells are inoculated in a 96-well plate, after overnight culture, compound protease is added for treatment (0 ug/ml, 10ug/ml, 25ug/ml, 50 ug/ml);
(2) OD measurements were made 48h after treatment using CCK-8 reagent and are shown in Table 1 and FIG. 1.
TABLE 1 CCK-8 OD measurements (statistical differences are shown in FIG. 1)
| 0μg/mL | 10μg/mL | 25μg/mL | 50μg/ |
|
| 24h | 0.323±0.021 | 0.346±0.023 | 0.444±0.013 | 0.448±0.019 |
| 48h | 0.485±0.022 | 0.528±0.021 | 0.725±0.028 | 0.792±0.019 |
From fig. 1 and table 1, it can be seen that the composite polypeptide prepared by the invention can obviously promote the proliferation of the porcine small intestine epithelial cell IPEC-J2, and the difference has statistical significance.
Example 3
(1) The IPEC-J2 cells are inoculated in a 6-well plate, when the cell density reaches 85%, a 200ul gun head is used for gently sliding over an adherent cell layer in the vertical direction of a culture hole, and PBS is used for removing detached cells;
(2) media containing 0. mu.g/mL and 50. mu.g/mL were added and photographed at 0h and 48h using an inverted microscope, respectively, and the results of the experiment are shown in FIG. 2.
As can be seen from FIG. 2, the composite polypeptide can significantly promote the migration ability of porcine small intestine epithelial cells IPEC-J2.
In conclusion, the composite polypeptide prepared by the invention can replace damaged cells by promoting the proliferation of the porcine small intestine epithelial cells on one hand, and cover the damaged area by promoting the migration of the porcine small intestine epithelial cells on the other hand, so that the composite polypeptide can be used for preparing a repairing agent for repairing the porcine intestinal tract injury to reduce the diarrhea of weaned piglets and improve the survival rate of the weaned piglets.
Claims (8)
1. The application of the composite polypeptide in preparing the pig intestinal tract repairing agent is characterized in that the composite polypeptide is prepared from the raw materials of long-forehead oplophorus macrobrachium shrimp meat and small-red oplophorus macrobrachium shrimp meat.
2. The use according to claim 1, wherein the complex polypeptide is prepared by the following method:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
3. The application of the composite polypeptide in preparing the promoter for promoting the proliferation of the epithelial cells of the small intestine of the pig is characterized in that the preparation method of the composite polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
4. The application of the composite polypeptide in preparing the promoter for promoting the migration of the epithelial cells of the small intestine of the pig is characterized in that the preparation method of the composite polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
5. The repairing agent for repairing the pig intestinal tract is characterized by being a composite polypeptide, wherein the composite polypeptide is prepared from prawns of red thorns.
6. The repair agent according to claim 5, wherein the complex polypeptide is prepared by the following method:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
7. An accelerant for promoting the proliferation of small intestinal epithelial cells of pigs is characterized in that the accelerant is a composite polypeptide, and the preparation raw materials of the composite polypeptide are prawns of red blood;
the preparation method of the composite polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
8. An accelerant for promoting migration of epithelial cells of small intestines of pigs is characterized in that the accelerant is a composite polypeptide, and the composite polypeptide is prepared from prawns of red thorns;
the preparation method of the composite polypeptide comprises the following steps:
(1) mixing the shrimp meat of the prawns with the shrimp meat of the small red prawns according to the weight ratio of 1: 1, grinding and mixing, adding 15 times of water to prepare homogenate, and sterilizing for later use;
(2) adding 3wt% of bacillus subtilis, fermenting at 35 ℃, pH6,30r/min and 48 h;
(3) sterilizing at high temperature and high pressure for 20min after fermentation;
(4) cooling to room temperature, and centrifuging the sterilized fermentation broth at 3500rpm for 20 min;
(5) taking the supernatant, and filtering with an ultrafiltration membrane of 1um to obtain polypeptide liquid;
(6) and (5) freeze-drying to obtain the composite polypeptide of the long-headed acanthomys prawns and the small red-headed acanthomys prawns.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011100442.5A CN112168950B (en) | 2020-10-15 | 2020-10-15 | Repairing agent for repairing pig intestinal tract |
| CN202210899469.8A CN115252756A (en) | 2020-10-15 | 2020-10-15 | Biological agent for reducing pig diarrhea |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011100442.5A CN112168950B (en) | 2020-10-15 | 2020-10-15 | Repairing agent for repairing pig intestinal tract |
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| CN202210899469.8A Division CN115252756A (en) | 2020-10-15 | 2020-10-15 | Biological agent for reducing pig diarrhea |
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| CN202011100442.5A Active CN112168950B (en) | 2020-10-15 | 2020-10-15 | Repairing agent for repairing pig intestinal tract |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117695370A (en) * | 2023-12-19 | 2024-03-15 | 青岛市畜牧工作站(青岛市畜牧兽医研究所) | Application of oligopeptide in preparation of porcine epidemic diarrhea virus inhibition drugs |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239960A (en) * | 2011-05-13 | 2011-11-16 | 湖北邦之德牧业科技有限公司 | Production method of biological polypeptide used for feed |
| CN107821821A (en) * | 2017-12-01 | 2018-03-23 | 无为县雨露生态农业有限公司 | Improve intestinal health and somatotrophic pig feed additive |
| CN110663803A (en) * | 2019-11-01 | 2020-01-10 | 付鹏磊 | Euphausia superba oil polypeptide biological product capable of benefiting spleen and invigorating stomach |
-
2020
- 2020-10-15 CN CN202210899469.8A patent/CN115252756A/en active Pending
- 2020-10-15 CN CN202011100442.5A patent/CN112168950B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102239960A (en) * | 2011-05-13 | 2011-11-16 | 湖北邦之德牧业科技有限公司 | Production method of biological polypeptide used for feed |
| CN107821821A (en) * | 2017-12-01 | 2018-03-23 | 无为县雨露生态农业有限公司 | Improve intestinal health and somatotrophic pig feed additive |
| CN110663803A (en) * | 2019-11-01 | 2020-01-10 | 付鹏磊 | Euphausia superba oil polypeptide biological product capable of benefiting spleen and invigorating stomach |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117695370A (en) * | 2023-12-19 | 2024-03-15 | 青岛市畜牧工作站(青岛市畜牧兽医研究所) | Application of oligopeptide in preparation of porcine epidemic diarrhea virus inhibition drugs |
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| CN112168950B (en) | 2022-08-02 |
| CN115252756A (en) | 2022-11-01 |
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