A nutrient solution containing chlorella, its preparation method, application and usage method

Abstract

The present invention relates to the technical field of livestock feed, and specifically relates to a nutrient solution containing chlorella, its preparation method, application and usage method. The nutrient solution includes: a chlorella product, a nutritional additive, and a rumen fermentation regulator. The ratio of the chlorella product, the nutritional additive, and the rumen fermentation regulator is 100 mL: (5 - 10) g: (20 - 35) g; the chlorella product is prepared from a chlorella enzymatic hydrolysate and a Lactobacillus plantarum fermentation broth; the nutritional additive includes vitamins and trace elements; the rumen fermentation regulator includes sodium bicarbonate and chitosan. The present invention also provides the application of the nutrient solution in the breeding of housed beef cattle, as well as the preparation and usage methods of the nutrient solution. The nutrient solution containing chlorella prepared by the present invention can not only improve the immunity of housed beef cattle, but also reduce the fat deposition in beef cattle and improve the quality of beef.

Description

Technical Field

[0001] The present invention belongs to the technical field of livestock feed, and particularly relates to a nutrient solution containing chlorella, a preparation method thereof, an application and a usage method. Background Art

[0002] Although the indoor feeding of beef cattle has advantages such as increasing the growth rate, improving the diet return, and reducing the grassland carrying capacity, it also brings some problems that cannot be ignored. Among them, the more prominent problem is that under the condition of high-concentrate diet, the intramuscular fat content of beef cattle increases. On the one hand, it leads to potential health hazards of beef cattle (decreased immunity), such as the occurrence of nutritional metabolic diseases such as fatty liver. On the other hand, it reduces the edible value of beef (decreased meat quality). At the same time, indoor-fed beef cattle also generally have problems such as decreased feed intake and low production performance.

[0003] From the perspective of enhancing immunity, improving meat quality and increasing feed intake, the current nutritional measures taken in the production of indoor-fed beef cattle mainly include: adding enzyme preparations, α-linolenic acid, organic acids, Chinese herbal medicine additives, adding trace element and vitamin additives, plant extracts, chitosan, and probiotics and other technologies. Among the above technologies, enzyme preparations, organic acids, chitosan, and probiotics are easily affected by the rumen fermentation characteristics of beef cattle, and the effects are not very stable; the application effect of Chinese herbal medicine additives is easily affected by raw materials, compatibility, and processing technology, and the price is relatively expensive; trace element and vitamin additives only consider the trace element and vitamin nutrition of beef cattle. Under normal circumstances, these two nutrients are not easily lacking in the mixed diet of beef cattle; α-linolenic acid needs to solve the problem of bypassing the rumen, otherwise the effect is mediocre; plant extracts are also easily affected by their sources, extraction processes, dosages, etc. Therefore, the application of such technologies in the actual production of indoor-fed beef cattle is also very limited.

[0004] Therefore, it is urgent to improve the existing nutritional measures for indoor-fed beef cattle and provide a nutrient solution to solve the problems of decreased meat quality and reduced immunity in the production of indoor-fed beef cattle. Summary of the Invention

[0005] To solve the technical problems of decreased meat quality and reduced immunity of indoor-fed beef cattle in the prior art, the present invention provides a nutrient solution containing chlorella, a preparation method thereof, an application and a usage method.

[0006] To achieve the above object, the technical scheme adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a nutrient solution containing chlorella, which comprises a chlorella product, a nutritional additive, and a rumen fermentation regulator. The ratio of the chlorella product, the nutritional additive, and the rumen fermentation regulator is 100 mL:(5 - 10) g:(20 - 35) g. The chlorella product is prepared from a chlorella enzymatic hydrolysate and a Lactobacillus plantarum fermentation broth. The nutritional additive includes vitamins and trace elements. The rumen fermentation regulator includes sodium bicarbonate and chitosan.

[0008] Preferably, the chlorella product is obtained by mixing the chlorella enzymatic hydrolysate and the Lactobacillus plantarum fermentation broth at a volume ratio of 5:(1 - 3), followed by low-pressure homogenization treatment for 10 - 15 min at 20 - 40 MPa.

[0009] Preferably, the preparation process of the chlorella enzymatic hydrolysate is as follows: Take 100 mL of chlorella culture solution, perform high-pressure homogenization treatment at 80 - 100 MPa for 10 min to obtain a chlorella cell homogenate. Add 10 - 20 g of wheat gluten, stir for 5 min to obtain a mixed solution. Add an alkaline protease accounting for 2 - 8% of the mass of the mixed solution, adjust the pH value to 8 - 10, and perform ultrasonic-assisted enzymatic hydrolysis in a water bath at 40 - 55 °C for 40 - 60 min to obtain an enzymatic hydrolysate.

[0010] Preferably, the Lactobacillus plantarum fermentation broth is prepared according to the following steps: Add the chlorella culture solution to the MRS liquid medium and perform high-temperature sterilization. After standing at room temperature, inoculate the Lactobacillus plantarum bacterial suspension and ferment at 37 °C for 8 - 12 h to obtain a fermentation broth. The volume ratio of the chlorella culture solution, the MRS liquid medium, and the Lactobacillus plantarum bacterial suspension is (0.1 - 0.3):1:0.08.

[0011] Preferably, the vitamins include nicotinic acid, biotin, and folic acid, and the trace elements include selenium, zinc, iron, copper, manganese, iodine, and cobalt.

[0012] Preferably, based on the total mass of the nutritional additive being 100%, the nutritional additive includes the following components in the following mass percentages: nicotinic acid 35 - 45%, biotin 1 - 3%, folic acid 5 - 9%, selenium 0.1 - 0.7%, zinc 11 - 14%, iron 22 - 32%, copper 1 - 3%, manganese 6 - 10%, iodine 0.1 - 0.5%, cobalt 0.1 - 0.5%.

[0013] Preferably, the mass ratio of sodium bicarbonate to chitosan in the rumen fermentation regulator is 100:(1 - 10).

[0014] In a second aspect, the present invention provides a preparation method of a nutrient solution containing chlorella, which comprises the following steps: Add the nutritional additive and the rumen fermentation regulator to the chlorella product in proportion and mix to obtain a nutrient solution containing chlorella.

[0015] Third aspect, the present invention provides an application of a nutrient solution containing chlorella, which is applied to the intensive feeding of beef cattle. Among them, each bull in the intensively fed beef cattle is supplied with 40 - 60 mL per day, and each cow in the intensively fed beef cattle is supplied with 70 - 100 mL per day.

[0016] Fourth aspect, the present invention provides a method for using a nutrient solution containing chlorella. The nutrient solution is used after being mixed with the feed of intensively fed beef cattle. The mixing ratio of the nutrient solution to the feed is 30 - 40 mL: 5 kg.

[0017] Integrating the above technical solutions, the present invention has the following beneficial effects:

[0018] (1) After the chlorella culture solution in the present invention is subjected to high-pressure homogenization cell disruption, rich nutritional components such as proteins, polysaccharides, and minerals inside the cells are released. In particular, the protein, which accounts for the highest mass percentage (about 60%) in chlorella, undergoes hydrolysis under the action of alkaline protease to generate peptide segments with different molecular weights. These peptide segments and other nutritional components are directly absorbed and utilized in the gastrointestinal tract. At the same time, the small peptides in these peptide segments can combine with Zn, Fe, Cu, etc. in the form of chelates and can promote the transport and absorption of these substances in cattle, so as to increase the content of trace elements in beef.

[0019] (2) The present invention simultaneously hydrolyzes chlorella protein and wheat protein in wheat gluten by alkaline protease and ultrasonic-assisted enzymatic hydrolysis method. The ultrasonic treatment makes the hydrolysis more sufficient. The chlorella protein peptides and wheat protein peptides in the hydrolysate jointly stimulate the secretion of growth hormone in beef cattle, thereby promoting the growth and development of beef cattle. It can not only increase the weight but also optimize the muscle fibers. During the growth process of beef cattle, an appropriate amount of plant peptides helps to form finer muscle fibers, making the beef taste better and having higher tenderness, thus improving the quality of beef.

[0020] (3) The present invention co-cultures Lactobacillus plantarum with chlorella culture solution and MRS liquid medium. The prepared Lactobacillus plantarum fermentation broth not only contains Lactobacillus plantarum and its metabolites, but also contains the nutritional components in chlorella. And after fermentation, the typical algae odor of chlorella is greatly reduced, improving the palatability of the nutrient solution. After supplying Lactobacillus plantarum and its metabolites (such as lactic acid, bacteriocin, hydrogen peroxide, amylase, protease, etc.) to intensively fed beef cattle, it can not only regulate the intestinal flora, promote digestion and absorption, but also enhance immunity and prevent diseases.

[0021] (4) The nutrient solution of the present invention also contains a rumen fermentation regulator. Among them, sodium bicarbonate can neutralize excessive acidic substances produced in the rumen, thereby maintaining the acid-base balance in the rumen and improving the digestive characteristics of the gastrointestinal tract of beef cattle. Chitosan activates the complement system, mediates a series of biological effects of the complement, and promotes the secretion of various immune factors by the bovine body, thereby enhancing the function of the body's immune system.

[0022] In summary, the nutrient solution containing Chlorella in the present invention contains various plant peptide components with different molecular weights, nutritional additives, and rumen fermentation regulators. They jointly provide the necessary nutritional components for the breeding of housed beef cattle, improving the immunity of housed beef cattle and the quality of beef. In addition, this technical solution is not only applicable to beef cattle, but can also be used to produce nutrient solutions for other ruminants, such as nutrient solutions for meat sheep or dairy cows, by changing the design. Description of the Drawings

[0023] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art.

[0024] Figure 1 It is a process flow diagram of the preparation method of a nutrient solution containing Chlorella in the present invention. Specific Embodiments

[0025] To further elaborate on the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following combines the attached Figure 1 and preferred embodiments to describe in detail the specific preparation method, components and their effects according to the present invention. It should be noted that the Chlorella in the Chlorella culture solution selected in this application is Chlorella vulgaris, and the Chlorella culture solution is prepared by a three-stage culture method. The specific preparation process is shown in the following specification section; the wheat gluten contains 70% wheat protein, and its manufacturer is Shandong Juxinlong Biotechnology Co., Ltd.; the enzyme activity of the alkaline protease is 100,000 U / g, and it is purchased from Weifang Ruichen Biotechnology Co., Ltd. The freeze-dried powder of Lactiplantibacillus plantarum is purchased from Xi'an Zebang Biotechnology Co., Ltd., and the product specification is 10 billion cfu / g.

[0026] The three-stage culture method for preparing the Chlorella culture solution specifically includes the following steps:

[0027] Step S1. Activate the Chlorella vulgaris and transfer it into a 1L BG-11 culture container with a volume of 2 / 3. There is an air vent at the bottom of the container. Pass gas into the culture container through the air vent. The flow rate of the gas is 2.5L / min. The cotton ball is disinfected before the gas enters the culture container. Cultivate for 8 days at a light intensity of 8000Lux and a culture temperature of 27°C to obtain the primary culture;

[0028] Step S2. Transfer the primary culture into a 50L BG-11 culture container with a volume of 2 / 3. There is an air vent at the bottom of the container. Pass gas into the culture container through the air vent. The flow rate of the gas is 3L / min. Cultivate for 10 days at a light intensity of 8000Lux and a culture temperature of 27°C to obtain the secondary culture;

[0029] Step S3. Transfer the secondary culture into a 185L BG-11 culture container with a volume of 2 / 3. There is an air vent at the bottom of the container. Pass gas into the culture container through the air vent. The flow rate of the gas is 3L / min. Cultivate for 22 days at a light intensity of 80000Lux and a culture temperature of 27°C to obtain the tertiary culture.

[0030] Example 1

[0031] This example provides a preparation method of a nutrient solution containing Chlorella vulgaris. Refer to the appendix Figure 1 , and the specific technical solution is as follows:

[0032] Step 1. Preparation of Chlorella vulgaris enzyme hydrolysate

[0033] Take 100mL of Chlorella vulgaris culture solution, perform high-pressure homogenization treatment at 90MPa for 12min to obtain Chlorella vulgaris cell homogenate. Add 15g of wheat gluten, stir for 5min to obtain a mixed solution. Add alkaline protease accounting for 5% (5.75g) of the mass of the mixed solution, adjust the pH value to 9, and perform ultrasonic-assisted enzymatic hydrolysis in a 48°C water bath for 50min to obtain the enzyme hydrolysate.

[0034] Step 2. Preparation of Lactobacillus plantarum fermentation broth

[0035] Prepare a Lactobacillus plantarum suspension: Open the ampoule bottle containing the freeze-dried powder of Lactobacillus plantarum in a super clean bench, mix the freeze-dried powder into a triangular flask containing 50mL of MRS liquid medium, and perform shake flask activation culture at 37°C for 24h to prepare a Lactobacillus plantarum suspension.

[0036] Into a 250 mL Erlenmeyer flask, add 100 mL of MRS liquid medium and 20 mL of Chlorella culture solution, stir for 5 min, seal the Erlenmeyer flask mouth, place it in an autoclave, sterilize at 0.1 MPa and 121 °C for 30 min, take it out from the autoclave, let it stand at room temperature and then inoculate 8 mL of Lactobacillus plantarum suspension, and ferment at 37 °C for 10 h to obtain a fermentation broth.

[0037] Step 3: Preparation of Chlorella products

[0038] Take 100 mL of Chlorella enzymolysis solution and 40 mL of Lactobacillus plantarum fermentation broth, perform low-pressure homogenization treatment at 30 MPa for 15 min to obtain Chlorella products.

[0039] Step 4: Preparation of nutrient solution

[0040] Take 100 mL of Chlorella products, add 8 g of nutritional additive and 28 g of rumen fermentation regulator, and mix evenly to obtain a nutrient solution containing Chlorella.

[0041] Among them, the nutritional additive includes the following components in mass percentage: nicotinic acid 40%, biotin 2%, folic acid 7%, selenium 0.4%, zinc 13%, iron 27%, copper 2%, manganese 8%, iodine 0.3%, cobalt 0.3%, and the trace elements are provided by sodium selenite, zinc sulfate, ferrous sulfate, copper sulfate, manganese sulfate, sodium iodate and cobalt sulfate; the rumen fermentation regulator contains 27 g of sodium bicarbonate and 1 g of chitosan.

[0042] Example 2

[0043] This example provides a preparation method of a nutrient solution containing Chlorella, and the specific technical solution is as follows:

[0044] Step 1: Preparation of Chlorella enzymolysis solution

[0045] Take 100 mL of Chlorella culture solution, perform high-pressure homogenization treatment at 80 MPa for 15 min to obtain a Chlorella cell homogenate, add 10 g of wheat gluten, stir for 5 min to obtain a mixed solution, add 2% (2.2 g) of alkaline protease based on the mass of the mixed solution, adjust the pH value to 10, and perform ultrasonic-assisted enzymolysis in a 40 °C water bath for 60 min to obtain an enzymolysis solution.

[0046] Step 2: Preparation of Lactobacillus plantarum fermentation broth

[0047] Prepare a Lactobacillus plantarum suspension: Open the ampoule bottle containing the freeze-dried powder of Lactobacillus plantarum in a superclean bench, mix the freeze-dried powder into a 50 mL Erlenmeyer flask containing MRS liquid medium, and perform shaking flask activation culture at 37 °C for 24 h to prepare a Lactobacillus plantarum suspension.

[0048] Into a 250 mL Erlenmeyer flask, add 100 mL of MRS liquid medium and 10 mL of Chlorella culture solution, stir for 5 min, seal the Erlenmeyer flask mouth, place it in an autoclave, sterilize at 0.1 MPa and 121 °C for 30 min, take it out from the autoclave, let it stand at room temperature and then inoculate 8 mL of Lactobacillus plantarum suspension, ferment at 37 °C for 8 h to obtain a fermentation broth.

[0049] Step 3: Preparation of Chlorella products

[0050] Take 100 mL of Chlorella enzymolysis solution and 20 mL of Lactobacillus plantarum fermentation broth, carry out low-pressure homogenization treatment at 20 MPa for 10 min to obtain Chlorella products.

[0051] Step 4: Preparation of nutrient solution

[0052] Take 100 mL of Chlorella products, add 5 g of nutrient additive and 35 g of rumen fermentation regulator, mix evenly to obtain a nutrient solution containing Chlorella.

[0053] Among them, the nutrient additive includes the following components in mass percentage: nicotinic acid 35%, biotin 3%, folic acid 5.7%, selenium 0.7%, zinc 14%, iron 32%, copper 3%, manganese 6%, iodine 0.1%, cobalt 0.5%, and the trace elements are provided by sodium selenite, zinc sulfate, ferrous sulfate, copper sulfate, manganese sulfate, sodium iodate and cobalt sulfate; the rumen fermentation regulator contains 31 g of sodium bicarbonate and 4 g of chitosan.

[0054] Example 3

[0055] This example provides a preparation method of a nutrient solution containing Chlorella, and the specific technical solution is as follows:

[0056] Step 1: Preparation of Chlorella enzymolysis solution

[0057] Take 100 mL of Chlorella culture solution, carry out high-pressure homogenization treatment at 100 MPa for 10 min to obtain a Chlorella cell homogenate, add 20 g of wheat gluten, stir for 5 min to obtain a mixed solution, add alkaline protease accounting for 8% (9.6 g) of the mass of the mixed solution, adjust the pH value to 8, and carry out ultrasonic-assisted enzymolysis in a 55 °C water bath for 40 min to obtain an enzymolysis solution.

[0058] Step 2: Preparation of Lactobacillus plantarum fermentation broth

[0059] Preparation of Lactobacillus plantarum suspension: Open the ampoule bottle containing Lactobacillus plantarum freeze-dried powder in a superclean bench, mix the freeze-dried powder into a 50 mL Erlenmeyer flask containing MRS liquid medium, and carry out shake flask activation culture at 37 °C for 24 h to prepare a Lactobacillus plantarum suspension.

[0060] Into an Erlenmeyer flask with a capacity of 250 mL, add 100 mL of MRS liquid medium and 30 mL of Chlorella culture solution, stir for 5 min, seal the mouth of the Erlenmeyer flask, place it in an autoclave, sterilize at 0.1 MPa and 121 °C for 30 min, take it out of the autoclave, let it stand at room temperature and then inoculate 8 mL of Lactobacillus plantarum suspension, and ferment at 37 °C for 12 h to obtain a fermentation broth.

[0061] Step 3: Preparation of Chlorella products

[0062] Take 100 mL of Chlorella enzymatic hydrolysate and 60 mL of Lactobacillus plantarum fermentation broth, perform low-pressure homogenization treatment at 40 MPa for 10 min to obtain Chlorella products.

[0063] Step 4: Preparation of nutrient solution

[0064] Take 100 mL of Chlorella products, add 10 g of nutrient additive and 20 g of rumen fermentation regulator, and mix evenly to obtain a nutrient solution containing Chlorella.

[0065] Among them, the nutrient additive includes the following components by mass percentage: nicotinic acid 45%, biotin 1%, folic acid 9%, selenium 0.1%, zinc 11%, iron 22%, copper 1%, manganese 10%, iodine 0.5%, cobalt 0.4%, and the trace elements are provided by sodium selenite, zinc sulfate, ferrous sulfate, copper sulfate, manganese sulfate, sodium iodate and cobalt sulfate; the rumen fermentation regulator contains 18 g of sodium bicarbonate and 2 g of chitosan.

[0066] Example 4

[0067] This example provides a preparation method of a nutrient solution containing Chlorella, and the specific technical solution is as follows:

[0068] Step 1: Preparation of Chlorella enzymatic hydrolysate

[0069] Take 100 mL of Chlorella culture solution, perform high-pressure homogenization treatment at 95 MPa for 10 min to obtain a Chlorella cell homogenate, add 20 g of wheat gluten, stir for 5 min to obtain a mixed solution, add alkaline protease accounting for 8% (9.6 g) of the mass of the mixed solution, adjust the pH value to 8, and perform ultrasonic-assisted enzymatic hydrolysis in a 50 °C water bath for 40 min to obtain an enzymatic hydrolysate.

[0070] Step 2: Preparation of Lactobacillus plantarum fermentation broth

[0071] Prepare a Lactobacillus plantarum suspension: Open the ampoule bottle containing the freeze-dried powder of Lactobacillus plantarum in a laminar flow hood, mix the freeze-dried powder into a triangular flask containing 50 mL of MRS liquid medium, and perform shaking flask activation culture at 37 °C for 24 h to prepare a Lactobacillus plantarum suspension.

[0072] Into a 250 mL Erlenmeyer flask, add 100 mL of MRS liquid medium and 30 mL of Chlorella vulgaris culture solution, stir for 5 min, seal the Erlenmeyer flask mouth, place it in an autoclave, sterilize at 0.1 MPa and 121 °C for 30 min, take it out from the autoclave, let it stand at room temperature and then inoculate 8 mL of Lactobacillus plantarum suspension, and ferment at 37 °C for 12 h to obtain a fermentation broth.

[0073] Step 3: Preparation of Chlorella vulgaris product

[0074] Take 100 mL of Chlorella vulgaris enzymatic hydrolysate and 60 mL of Lactobacillus plantarum fermentation broth, perform low-pressure homogenization treatment at 35 MPa for 10 min to obtain a Chlorella vulgaris product.

[0075] Step 4: Preparation of nutrient solution

[0076] Take 100 mL of Chlorella vulgaris product, add 10 g of nutrient additive and 20 g of rumen fermentation regulator, and mix evenly to obtain a nutrient solution containing Chlorella vulgaris.

[0077] Among them, the nutrient additive includes the following components in mass percentages: nicotinic acid 40%, biotin 2%, folic acid 5%, selenium 0.4%, zinc 13%, iron 27%, copper 2%, manganese 10%, iodine 0.5%, cobalt 0.1%, and the trace elements are provided by sodium selenite, zinc sulfate, ferrous sulfate, copper sulfate, manganese sulfate, sodium iodate and cobalt sulfate; the rumen fermentation regulator contains 18 g of sodium bicarbonate and 2 g of chitosan.

[0078] It should be noted that in Examples 1-4, the order of Step 1 and Step 2 can be interchanged, which does not affect the preparation and use effect of the nutrient solution.

[0079] Comparative Example 1

[0080] The difference between this comparative example and Example 1 is that when preparing the Chlorella vulgaris enzymatic hydrolysate in Step 1, wheat gluten is not added.

[0081] Comparative Example 2

[0082] The difference between this comparative example and Example 1 is that when preparing the Lactobacillus plantarum fermentation broth in Step 2, Chlorella vulgaris culture solution is not added to the MRS medium.

[0083] Comparative Example 3

[0084] The difference between this comparative example and Example 1 is that when preparing the nutrient solution in Step 4, the rumen fermentation regulator does not contain sodium bicarbonate.

[0085] Comparative Example 4

[0086] The difference between this comparative example and Example 1 is that when preparing the nutrient solution in Step 4, the rumen fermentation regulator does not contain chitosan.

[0087] Performance Test

[0088] To explore the effects of the nutrient solution containing Chlorella prepared by the present invention on the immune function of housed beef cattle and the quality of their meat and fat, 45 housed Horqin bulls with a body weight of 380 to 400 kg were selected as the animal model. They were randomly divided into 9 groups of 5 cows each, namely a control group and 8 experimental groups.

[0089] The blank control group was fed a basal diet. The 8 experimental groups were respectively added with 50 mL / head·day of the nutrient solution (the nutrient solutions in the 8 experimental groups were the nutrient solutions prepared by the methods in Examples 1-4 and Comparative Examples 1-4) to the basal diet at a ratio of 7 mL:1 kg. A feeding comparison test was carried out for 70 days, among which the 1st to 10th days were the pre-test period, and the 11th to 70th days were the test period. During the pre-test period, numbering, deworming, and stomach-strengthening were carried out. The deworming drug was ivermectin, and after deworming, rhubarb and sodium bicarbonate powder were used for stomach-strengthening. At the beginning (the 11th day) and the end (the 70th day) of the test period, jugular vein blood samples were taken from each group of test cattle and serum was separated for detecting the contents of immune indexes lysozyme (LZM), nitric oxide (NO), acid phosphatase (ACP), and immunoglobulin G (IgG). The detection results of the immune indexes in the serum of each group of cattle (the average value of the detection of 5 cows in each group was used as the detection result) are shown in Table 1.

[0090] Table 1 Detection of Immune Indexes in Bovine Serum

[0091]

[0092]

[0093] According to the detection results of the immune indexes in bovine serum in Table 1, compared with Comparative Examples 1-4 and the blank control example, the contents of immune indexes lysozyme, nitric oxide, acid phosphatase, and immunoglobulin G in the serum of cattle in the experimental groups using the nutrient solutions prepared in Examples 1-4 were greatly improved, indicating that the nutrient solution provided in the examples of the present invention can significantly improve the immune performance of housed beef cattle. At the same time, the immune indexes in the serum of cattle in the blank control group fed only the basal diet were the lowest, indicating that only relying on the basal diet cannot meet the immune requirements of housed beef cattle and the immune function of the cattle body is low.

[0094] After the experiment, one cow was randomly slaughtered from each group, and 30 g of the longissimus dorsi muscle samples were taken for the detection and analysis of fat content. The detection process of fat is as follows: The 30 g sample was minced, and the minced sample was placed in a 250 mL conical flask. 100 mL of 2 mol / L hydrochloric acid solution was added, and it was heated to boiling to free the fat in the meat. The filter residue containing fat was obtained by filtration. The filter residue and the filter paper were placed in an oven to dry to remove moisture. The dried filter paper and filter residue were put into a Soxhlet extractor, and 100 mL of petroleum ether was added as the extractant. The fat in the filter residue was extracted by the Soxhlet extraction method. After the extraction was completed, the extractant was evaporated to obtain pure fat, and the weight was taken to obtain the fat content in the longissimus dorsi muscle sample. The detection results of the fat content in the longissimus dorsi muscle of cattle are shown in Table 2.

[0095] Table 2 Detection results of the fat content in the longissimus dorsi muscle of cattle

[0096]

[0097]

[0098] As can be seen from Table 2, the fat content in the longissimus dorsi muscle of cattle in Examples 1 - 4 was significantly lower than that in Comparative Examples 1 - 4 and the blank control, indicating that the nutrient solution provided by the present invention can reduce the fat deposition in stall-fed beef cattle and improve the beef quality, thereby avoiding diseases such as fatty liver caused by excessive fat deposition.

[0099] In addition to using stall-fed Horqin bulls as animal models for performance testing as above, the inventors also conducted performance testing with stall-fed Horqin cows as animal models and found that the nutrient solution prepared by the present invention is also applicable to cows and can also improve the immune performance and beef quality of cows.

[0100] In addition, based on the nutrient solution containing Chlorella provided by the present invention, nutrient solutions for other ruminants can be further prepared after formula optimization.

[0101] Based on the above technical solutions, a nutrient solution containing Chlorella provided by the present invention contains various plant peptide components with different molecular weights, nutritional additives, and rumen fermentation regulators. They jointly provide the required nutritional components for the breeding of stall-fed beef cattle, reduce fat deposition, and improve the immunity and beef quality of stall-fed beef cattle.

[0102] The above are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Although the present invention has been disclosed above with the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some changes or modifications to equivalent embodiments of equivalent changes by using the technical content disclosed above within the scope of the technical solution of the present invention. However, as long as it does not depart from the content of the technical solution of the present invention, any brief modifications, equivalent changes and modifications made to the above embodiments according to the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A nutrient solution containing chlorella, characterized in that, The nutrient solution includes a chlorella product, a nutritional additive, and a rumen fermentation regulator. The ratio of the chlorella product, the nutritional additive, and the rumen fermentation regulator is 100 mL : (5 - 10) g : (20 - 35) g. The chlorella product is prepared from a chlorella enzymatic hydrolysate and a Lactobacillus plantarum fermentation broth. The nutritional additive includes vitamins and trace elements. The rumen fermentation regulator includes sodium bicarbonate and chitosan. The preparation process of the chlorella enzymatic hydrolysate is as follows: Take 100 mL of chlorella culture solution, perform high-pressure homogenization treatment at 80 - 100 MPa for 10 min to obtain a chlorella cell homogenate. Add 10 - 20 g of wheat gluten, stir for 5 min to obtain a mixture. Add an alkaline protease accounting for 2 - 8% of the mass of the mixture, adjust the pH value to 8 - 10, and perform ultrasonic-assisted enzymatic hydrolysis in a water bath at 40 - 55 °C for 40 - 60 min to obtain an enzymatic hydrolysate. The Lactobacillus plantarum fermentation broth is prepared according to the following steps: Add the chlorella culture solution to the MRS liquid medium and perform high-temperature sterilization. After standing at room temperature, inoculate with a Lactobacillus plantarum bacterial suspension and ferment at 37 °C for 8 - 12 h to obtain a fermentation broth. The volume ratio of the chlorella culture solution, the MRS liquid medium, and the Lactobacillus plantarum bacterial suspension is (0.1 - 0.3) : 1 : 0.

08.

2. The nutrient solution containing chlorella according to claim 1, characterized in that, The chlorella product is obtained by mixing the chlorella enzymatic hydrolysate and the Lactobacillus plantarum fermentation broth according to a volume ratio of 5 : (1 - 3) and performing low-pressure homogenization treatment for 10 - 15 min. The low-pressure homogenization treatment is carried out at 20 - 40 MPa.

3. The nutrient solution containing chlorella according to claim 1, characterized in that, The vitamins include nicotinic acid, biotin, and folic acid. The trace elements include selenium, zinc, iron, copper, manganese, iodine, and cobalt.

4. A nutrient solution containing chlorella according to claim 3, characterized in that, Based on the total mass of the nutritional additive being 100%, the nutritional additive includes the following components in mass percentages: nicotinic acid 35 - 45%, biotin 1 - 3%, folic acid 5 - 9%, selenium 0.1 - 0.7%, zinc 11 - 14%, iron 22 - 32%, copper 1 - 3%, manganese 6 - 10%, iodine 0.1 - 0.5%, cobalt 0.1 - 0.5%.

5. A nutrient solution containing chlorella according to claim 1, characterized in that, The mass ratio of sodium bicarbonate and chitosan in the rumen fermentation regulator is 100 : (1 - 10).

6. A method for preparing a nutrient solution containing chlorella according to any one of claims 1-5, characterized in that, It includes the following steps: Add the nutritional additive and the rumen fermentation regulator to the chlorella product in proportion and mix to obtain a nutrient solution containing chlorella.

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