Functional feed for improving immunity of Macrobrachium rosenbergii and preparation method thereof

Abstract

The present invention discloses a functional feed for improving the immunity of Macrobrachium rosenbergii and a preparation method thereof. First, a feed additive is prepared, which is prepared by fermenting a composite microorganism using jujube kernel powder as a raw material; the composite microorganism is a mixture of plant lactobacillus and licheniformis in equal proportions. The preparation method comprises the following steps: adding 2.5 to 3 times the mass of water to the jujube kernel powder to prepare a suspension, adding the composite microorganism to the suspension at a ratio of 3 to 4‰, fermenting for 12 to 18 hours under a constant temperature of 30 to 37°C, adding tea water extract to mix, and grinding the solid powder obtained after drying to obtain the feed additive. Then, a functional feed for improving the immunity of Macrobrachium rosenbergii is prepared with a replacement rate of the feed additive in the basic feed of 5 to 10%, which can greatly improve the immunity and growth performance of Macrobrachium rosenbergii.

Description

Technical Field

[0001] The invention belongs to the technical field of aquatic animal breeding, and particularly relates to a functional feed for improving the immunity of Macrobrachium rosenbergii and a preparation method thereof. Background Art

[0002] Macrobrachium rosenbergii is large in size, has a wide range of diet, grows fast, and is rich in nutrition. The edible part contains 20.5% protein, which is equivalent to that of Chinese shrimp, and 1.97% fat, which is slightly higher than that of Chinese shrimp. Macrobrachium rosenbergii is delicious and is loved by consumers. It is farmed on a large scale in many areas. However, Macrobrachium rosenbergii is very prone to diseases during the breeding process, such as black spot disease and black gill disease, which seriously affect the quality and yield of Macrobrachium rosenbergii. At present, the common method for preventing and controlling black spot disease is to soak shrimp in a mixture of 0.9 mg / L malachite green and 2 mg / L formalin for 15 to 30 minutes for prevention when stocking; after the disease occurs, 1 mg / L furazolidone is sprayed throughout the pond for prevention and control. The method for preventing and controlling black gill disease is generally to soak in 0.2 mg / L malachite green for 30 minutes or in 0.4 mg / L copper sulfate solution for 6 hours, and each course of treatment lasts 3 to 5 days. These prevention and control methods are all chemical prevention and control, which cause great pollution to water bodies. Summary of the invention

[0003] In view of this, the present invention provides a functional feed for improving the immunity of Macrobrachium rosenbergii and a preparation method thereof. The additive is prepared by fermenting jujube pit powder with probiotics, and the additive is added to the basic feed of Macrobrachium rosenbergii to achieve the effect of improving the immunity of Macrobrachium rosenbergii, thereby effectively preventing and controlling diseases and increasing production.

[0004] In order to achieve the above object, the present invention adopts the following technical solutions:

[0005] The technical solution of the present invention is to provide a feed additive, which is prepared by composite microbial fermentation with jujube kernel powder as raw material; the composite microorganism is a mixture of Lactobacillus plantarum and Bacillus licheniformis in equal proportions.

[0006] As a further improvement of the present invention, the jujube core powder is a jujube core, and the particle size of the jujube core powder is less than 50 meshes. The specific preparation method is: remove the pulp of dry jujube, add water to steam and wash until the residual pulp on the core is completely removed, dry, crush, remove the jujube kernel, burn the jujube shell red, and grind it to a particle size of less than 50 meshes. The variety of jujube in the technical solution of the present invention is not limited.

[0007] Jujube kernel powder contains rich nutrients such as protein, fat, carbohydrates, vitamins and minerals. Among them, the protein content in jujube kernel is relatively high, which can provide amino acids. In addition, jujube kernels are also rich in minerals such as calcium, iron, and zinc. The present invention finds that jujube kernel powder has a good effect on improving the immune system of Macrobrachium rosenbergii. After fermenting jujube kernel powder with probiotics, part of the sugar can be removed, and at the same time, the probiotics metabolize to produce a large amount of beneficial enzymes and proteins, which have a good promoting effect on the digestion and metabolism of Macrobrachium rosenbergii, and further prevent the occurrence of diseases.

[0008] The technical solution of the second aspect of the present invention is to provide a method for preparing a feed additive, comprising adding 2.5 to 3 times the mass of water to the jujube kernel powder to prepare a suspension, adding the composite microorganism to the suspension at a ratio of 3 to 4‰ of the inoculation amount, fermenting for 12 to 18 hours under a constant temperature condition of 30 to 37°C, adding tea water extract to mix, and grinding the solid powder obtained after drying to obtain the feed additive.

[0009] Tea water extract is weakly alkaline. The present invention has found that adding an appropriate amount of tea water extract can appropriately increase the alkalinity of feed additives. On the other hand, by adding tea water extract to Macrobrachium rosenbergii feed, the nutritional value of the feed and the health level of Macrobrachium rosenbergii can be improved. Tea polyphenols have antioxidant, anti-inflammatory and antibacterial effects, can enhance the shelf life of feed, and help improve the growth rate and immunity of Macrobrachium rosenbergii.

[0010] As a further improvement of the present invention, the drying method is freeze-drying.

[0011] As a further improvement of the present invention, the amount of the tea water extract added is 25% to 30% based on the volume of the suspension.

[0012] As a further improvement of the present invention, the tea is green tea. The present invention compares the experimental effects of black tea, white tea and green tea and finds that green tea is most beneficial to the growth of Macrobrachium rosenbergii, and feed with green tea water extract has the best palatability, which is conducive to Macrobrachium rosenbergii feeding.

[0013] Preparation method of tea water extract: add 10 times the mass of water to green tea leaves, boil and then naturally cool to room temperature to obtain the extract.

[0014] The technical solution of the third aspect of the present invention is to provide the use of the above feed additive in Macrobrachium rosenbergii feed.

[0015] The technical solution of the fourth aspect of the present invention is to provide a functional feed for improving the immunity of Macrobrachium rosenbergii, wherein the above-mentioned feed additive partially replaces the basic feed, and the replacement rate is 5-10%. Since the most important function of feed additives is to improve disease resistance, the balance and comprehensiveness of its nutritional components are not as good as the basic feed, so the replacement rate of the basic feed should not be too high. Specifically, different proportions of replacement are required according to the growth stage of Macrobrachium rosenbergii. The replacement rate in juvenile shrimp feed is 5-7%, the replacement rate in middle shrimp feed is 7-8.5%, and the replacement rate in adult shrimp feed is 8.5-10.

[0016] As a further improvement of the present invention, the replacement rate in juvenile shrimp feed is 7%, the replacement rate in middle shrimp feed is 8.5%, and the replacement rate in adult shrimp feed is 10%.

[0017] The technical solution of the fifth aspect of the present invention is to provide a method for preparing a functional feed for improving the immunity of Macrobrachium rosenbergii, which comprises adding the above feed additives to the basic feed in proportion, mixing, and drying.

[0018] The technical effects of the present invention are as follows:

[0019] The feed additive of the present invention uses jujube kernel powder as a raw material. The rich flavonoids contained in the jujube kernel powder have antioxidant activity and can improve the immunity of Macrobrachium rosenbergii. In addition, the jujube kernel powder also contains rich nutrients such as protein, polysaccharides and minerals, which can enrich the nutrition of Macrobrachium rosenbergii. The sugar content in the jujube kernel powder fermented with probiotics is reduced to avoid the excessive sugar content affecting the appetite of Macrobrachium rosenbergii. At the same time, the content of nutrients such as protein is increased. Under the synergistic effect of various nutrients, the immunity and disease resistance of Macrobrachium rosenbergii are greatly improved. Moreover, the feed additive of the present invention is low in price and simple in preparation method.

[0020] The feed additive is applied to the feed of Macrobrachium rosenbergii. On the one hand, the nutritional value of the basic feed is enriched through the appropriate addition amount, which greatly improves the growth performance of Macrobrachium rosenbergii. On the other hand, the special ingredients and nutritional ingredients in the additive further improve the immunity of Macrobrachium rosenbergii. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The drawings constituting a part of the present application are used to provide a further understanding of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application.

[0022] Figure 1 This is a comparison chart of the effects of Macrobrachium rosenbergii raised in Application Example 2 of the present invention and Macrobrachium rosenbergii raised in the blank control group. DETAILED DESCRIPTION

[0023] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0024] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0025] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0026] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0027] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0028] The "room temperature" mentioned in the present invention is 25±2°C unless otherwise specified.

[0029] The invention provides a feed additive, which is prepared by composite microbial fermentation with jujube pit powder as a raw material; the composite microorganism is a mixture of plant lactobacillus and bacillus licheniformis in equal proportions.

[0030] In some preferred embodiments, the jujube pit powder is jujube pits, and the particle size of the jujube pit powder is less than 50 mesh.

[0031] The present invention also provides a method for preparing a feed additive, comprising adding 2.5 to 3 times the mass of water to the jujube kernel powder to prepare a suspension, adding the composite microorganism to the suspension at a ratio of 3 to 4‰, fermenting for 12 to 18 hours under a constant temperature condition of 30 to 37°C, adding tea water extract to mix, and grinding the solid powder obtained after drying to obtain the feed additive.

[0032] Tea water extract is weakly alkaline. The present invention has found that adding an appropriate amount of tea water extract can appropriately increase the alkalinity of feed additives. On the other hand, by adding tea water extract to Macrobrachium rosenbergii feed, the nutritional value of the feed and the health level of Macrobrachium rosenbergii can be improved. Tea polyphenols have antioxidant, anti-inflammatory and antibacterial effects, can enhance the shelf life of feed, and help improve the growth rate and immunity of Macrobrachium rosenbergii.

[0033] In some preferred embodiments of the present invention, the drying method is freeze-drying. The specific freeze-drying method can be conventional freeze-drying technology, which will not be described in detail here.

[0034] In some preferred embodiments of the present invention, the amount of the tea water extract added is 25% to 30% based on the volume of the suspension. The typical but non-limiting amount selected in the embodiments of the present invention is 30%.

[0035] In some preferred embodiments of the present invention, the tea is green tea. The present invention found through comparison of the experimental effects of black tea, white tea and green tea that green tea is most beneficial to the growth of Macrobrachium rosenbergii, and the feed added with green tea water extract has the best palatability, which is conducive to Macrobrachium rosenbergii feeding. Based on price considerations, the green tea selected in the embodiments of the present invention is low-grade Tieguanyin tea.

[0036] The invention also provides application of the feed additive in Macrobrachium rosenbergii feed.

[0037] The present invention also provides a functional feed for improving the immunity of Macrobrachium rosenbergii, wherein the above feed additive partially replaces the basic feed, and the replacement rate is 5-10%. Since the main function of feed additives is to improve disease resistance, the balance and comprehensiveness of its nutritional components are not as good as those of the basic feed, so the replacement rate of the basic feed should not be too high. Specifically, different proportions of replacement are required according to the growth stage of Macrobrachium rosenbergii. The replacement rate in juvenile shrimp feed is 5-7%, the replacement rate in middle shrimp feed is 7-8.5%, and the replacement rate in adult shrimp feed is 8.5-10.

[0038] The preparation method of jujube kernel powder in the following embodiment of the present invention is as follows: remove the pulp of dry jujube, add water to steam and wash until the residual pulp on the kernel is completely removed, dry, crush, remove the jujube kernel, burn the jujube shell red, and grind it to pass through a 50-mesh sieve. The variety of jujube in the technical scheme of the present invention is not limited. Based on price considerations, the jujube in the embodiment is selected from low-grade, cheap, less fleshy, large kernels and poor sweetness.

[0039] In the following embodiments of the present invention, the method for preparing the tea water extract is as follows: green tea leaves are added with 10 times the mass of water, boiled and then naturally cooled to room temperature to obtain the tea water extract.

[0040] In the embodiments of the present invention, Lactobacillus plantarum and Bacillus licheniformis were purchased from Xian Qianyecao Biotechnology Co., Ltd., with a content of ≥10 billion cfu / g (%); Bacillus licheniformis agent was purchased from Shanxi Xinyu Biotechnology Co., Ltd., with a content of ≥10 billion cfu / g (%).

[0041] The purchased bacterial agent is bacterial powder, which needs to be prepared into bacterial liquid before use. The method of preparing bacterial liquid is as follows:

[0042] (1) Sterilize the pipette tip and draw up 0.5 mL of ddHO. 2 O was added to the centrifuge tube containing Bacillus licheniformis, and gently shaken to form a bacterial suspension. 0.2 mL of the bacterial suspension was streaked and inoculated onto MRS solid medium, and activated and cultured at 35°C for 36 h.

[0043] (2) Use an inoculation loop to take 5 loops of activated Bacillus licheniformis and inoculate them into a 500 mL Erlenmeyer flask containing 150 mL of MRS liquid culture medium. Incubate the culture at 35°C on a rotary shaker (150 rpm) for 36 h. Then, expand the culture step by step according to the production volume to produce a production culture solution (the number of viable bacteria in the culture solution is controlled at 10 8 order of magnitude).

[0044] (3) The plant lactobacillus slant strain was streaked into MRS solid medium and cultured at 35°C for 36 h until a single colony grew. Five loops were taken with an inoculation loop and inoculated into a 500 mL Erlenmeyer flask containing 150 mL MRS liquid medium. The culture was expanded at 35°C for 36 h, and then the culture was expanded step by step according to the production volume to prepare a production culture solution (the number of viable bacteria in the culture solution was controlled at 10 8 order of magnitude).

[0045] The basal feed for Macrobrachium rosenbergii used in the embodiments of the present invention is prepared in strict accordance with the industry standard SC / T 1066-2003 compound feed. The main nutritional indicators are shown in Table 1.

[0046] Table 1 Main nutritional components of Macrobrachium rosenbergii feed (%)

[0047]

[0048] Example 1

[0049] A feed additive is prepared by composite microbial fermentation using jujube kernel powder as a raw material; the composite microorganism is fermented by mixing plant lactobacillus liquid and licheniformis liquid in equal proportions. The inoculation amount during the fermentation process is 3‰ (ml / kg), and the fermentation conditions are: 35°C, constant temperature sealed fermentation for 18 hours. After the fermentation is completed, tea water extract is added in an amount of 25%. After the mixture is evenly stirred, it is freeze-dried to prepare a feed additive.

[0050] Example 2

[0051] A feed additive is prepared by composite microbial fermentation using jujube kernel powder as a raw material; the composite microorganism is fermented by mixing plant lactobacillus liquid and licheniformis liquid in equal proportions. The inoculation amount during the fermentation process is 3.5‰ (ml / kg), and the fermentation conditions are: 35°C, constant temperature sealed fermentation for 15 hours. After the fermentation is completed, tea water extract is added in an amount of 25%. After the mixture is evenly stirred, it is freeze-dried to prepare a feed additive.

[0052] Example 3

[0053] A feed additive is prepared by composite microbial fermentation using jujube kernel powder as a raw material; the composite microorganism is fermented by mixing plant lactobacillus liquid and licheniformis liquid in equal proportions. The inoculation amount during the fermentation process is 4‰ (ml / kg), and the fermentation conditions are: 35°C, constant temperature sealed fermentation for 12 hours. After the fermentation is completed, tea water extract is added in an amount of 25%. After the mixture is evenly stirred, it is freeze-dried to prepare a feed additive.

[0054] Examples 1 to 3 investigated feed additives prepared with different inoculation amounts. The total flavonoid content of the feed additives prepared by the preparation methods of Examples 1 to 3 was detected by spectrophotometry, and it was found that the total flavonoid content of Example 2 was the highest. Therefore, the subsequent experiments were conducted based on Example 2.

[0055] Example 4

[0056] The only difference from Example 2 is that the amount of tea water extract added is 28%.

[0057] Example 5

[0058] The only difference from Example 2 is that the amount of tea water extract added is 30%.

[0059] Comparative Example 1

[0060] The only difference from Example 2 is that the amount of tea water extract added is 35%.

[0061] Application Examples

[0062] The immunity test of the present invention takes disease resistance and serum immunity-related enzyme activities as test objects.

[0063] The experiment was carried out at a certain artificial breeding base for Macrobrachium rosenbergii.

[0064] 400 healthy juvenile Macrobrachium rosenbergii were selected, and the size was required to be uniform. They were divided into 4 groups, each with 200 shrimps. The feed additives prepared in Examples 2, 4, 5 and Comparative Example 1 partially replaced the basic feed of Macrobrachium rosenbergii to prepare a compound feed. The specific replacement rate in the juvenile shrimp feed was 5%, the replacement rate in the middle shrimp feed was 7%, and the replacement rate in the adult shrimp feed was 9%. The feeding method and management method can be conventional management methods. After 90 days, they were raised to adult shrimps, and 10 shrimps were randomly selected from each group to test the activity of serum immune-related enzymes. The average results are shown in Table 2. 30 shrimps were randomly selected from each group and artificially infected with pathogenic Aeromonas hydrophila. The statistical mortality rate is shown in Table 3.

[0065] Table 2

[0066]

[0067] Table 3

[0068] Number of deaths (last) mortality rate(%) Example 2 1 3.33 Example 4 0 0 Example 5 2 6.67 Comparative Example 1 5 16.67

[0069] By comparing the data in Table 2 and Table 3, it can be seen that the feed additive in Example 4 has the best effect on improving the immunity of Macrobrachium rosenbergii. Therefore, the additive in Example 4 was selected for subsequent experiments.

[0070] Application Example 2

[0071] The feed additive prepared in Example 4 partially replaced the basic feed of Macrobrachium rosenbergii to prepare a compound feed. Specifically, the replacement rate in the juvenile shrimp feed was 6%, the replacement rate in the middle shrimp feed was 8.5%, and the replacement rate in the adult shrimp feed was 9.5%. The same method as in Application Example 1 was used for feeding and management, and the shrimps were raised to adult after 90 days.

[0072] Application Example 3

[0073] The feed additive prepared in Example 4 partially replaced the basic feed of Macrobrachium rosenbergii to prepare a compound feed. Specifically, the replacement rate in the juvenile shrimp feed was 7%, the replacement rate in the middle shrimp feed was 8.5%, and the replacement rate in the adult shrimp feed was 10%. The same method as in Application Example 1 was used for feeding and management, and the shrimps were raised to adult after 90 days.

[0074] Application Example 4

[0075] The feed additive prepared in Example 4 partially replaced the basic feed of Macrobrachium rosenbergii to prepare a compound feed. Specifically, the replacement rate in the juvenile shrimp feed was 8.5%, the replacement rate in the middle shrimp feed was 10%, and the replacement rate in the adult shrimp feed was 15%. The same method as in Application Example 1 was used for feeding and management, and the shrimps were raised to adult after 90 days.

[0076] 100 shrimps were randomly selected from each group, weighed and measured for body length. The average values ​​of total weight and body length are shown in Table 4.

[0077] Table 4

[0078] Total weight (g) Average body length (cm) Application Example 1 4431 11.7 Application Example 2 4683 11.9 Application Example 3 4211 11.4 Application Example 4 3823 10.9 Blank control 2122 9.1

[0079] Note: The replacement rate of additives in the basic feed in the blank control group is 0.

[0080] It can be seen from the data in Table 4 that although the feed additive of the present invention can improve immunity and growth performance to a certain extent, the more the amount of addition is, the better it is, as can be seen from the data comparison between Application Example 4 and the blank control. Among them, Application Example 2 has the best effect, that is, the most preferred replacement rate of the feed additive of the present invention in the feeding process is specifically: the replacement rate in juvenile shrimp feed is 6%, the replacement rate in middle shrimp feed is 8.5%, and the replacement rate in adult shrimp feed is 9.5%. Figure 1 Comparative photos of the largest Macrobrachium rosenbergii selected from Application Example 2 (left) and the blank control group (right).

[0081] The above are only preferred specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A feed additive, characterized in that: The method is prepared by using jujube pit powder as raw material and fermenting with composite microorganisms; the composite microorganisms are composed of a mixture of Lactobacillus plantarum and Bacillus licheniformis in equal proportions; The jujube kernel powder is jujube kernels, and the particle size of the jujube kernel powder is less than 50 meshes; Add 2.5 to 3 times the mass of water to the jujube kernel powder to prepare a suspension, add the composite microorganism to the suspension at an inoculum amount of 3 to 4‰ in ml / kg, ferment for 12 to 18 hours at a constant temperature of 30 to 37°C, add tea water extract to mix, dry and obtain a solid powder, and grind to obtain the feed additive; The amount of the tea water extract added is 25% to 30% based on the volume of the suspension; The preparation method of the tea water extract is as follows: adding 10 times the mass of water to green tea leaves, boiling the leaves, and then naturally cooling the leaves to room temperature to obtain the tea water extract.

2. The feed additive according to claim 1, characterized in that The drying method is freeze-drying.

3. A functional feed for improving the immunity of Macrobrachium rosenbergii, characterized in that: The basic feed is partially replaced by the feed additive according to any one of claims 1 to 2, with a replacement rate of 5 to 10%.

4. A method for preparing a functional feed for improving the immunity of Macrobrachium rosenbergii, comprising adding the feed additive according to any one of claims 1 to 2 to a basic feed in proportion, mixing, and drying.

Images