Auxiliary agent for treating or preventing fatty liver of carnivorous fish and preparation method thereof

Abstract

The present application relates to the field of fish feed, and more specifically, to an adjuvant for treating or preventing fatty liver in carnivorous fish and a preparation method thereof. An adjuvant for treating or preventing fatty liver in carnivorous fish comprises the following raw materials: 5000-7000 ug / kg vitamin B 12 , 500-750mg / kg vitamin C, 500-600mg / kg astaxanthin, 20000-30000mg / kg taurine, 150-200mg / kg chromium picolinate (calculated as chromium), 1500-3000KU / kg aflatoxin degrading enzyme, 1-10g / kg 2-deoxy-D-glucose; the carrier supplements the remainder. The application has the advantage of improving the formation of fatty liver in carnivorous fish.

Description

Technical Field

[0001] The present application relates to the field of fish feed, and more specifically, to an adjuvant for treating or preventing fatty liver of carnivorous fish and a preparation method thereof. Background Art

[0002] The liver is the largest metabolic organ in fish. Damage or disease to the liver often leads to metabolic disorders in fish, reduced disease resistance, and serious threats to intensive aquaculture. Nutritional fatty liver in fish caused by metabolic disorders has a high incidence in intensive aquaculture, especially in carnivorous fish.

[0003] Under natural conditions, carnivorous fish mainly eat meat, with protein, fat, etc. as their main energy sources, and their ability to utilize carbohydrates is weaker than that of herbivorous and omnivorous fish. In intensive farming, fish need to be fed pelleted feed, and 10-20% starch (sugar) is added to make the feed pelletized. Long-term intake of starch will cause sugar intolerance in carnivorous fish, causing a burden on the liver. Sugar cannot be metabolized and converted into fat, and accumulates in the liver, causing fatty liver. The problem of fatty liver in carnivorous fish is an important problem that restricts the development of the industry, and there is currently no good solution. Therefore, there is still room for improvement. Summary of the invention

[0004] In order to improve the formation of fatty liver in carnivorous fish, the present application provides an adjuvant for treating or preventing fatty liver in carnivorous fish and a preparation method thereof.

[0005] In a first aspect, the present application provides an adjuvant for treating or preventing fatty liver in carnivorous fish, using the following technical solution:

[0006] An auxiliary agent for treating or preventing fatty liver of carnivorous fish, comprising the following raw materials: 5000-7000ug / kg vitamin B 12 , 500-750mg / kg vitamin C, 500-600mg / kg astaxanthin, 20000-30000mg / kg taurine, 150-200mg / kg chromium picolinate (calculated as chromium), 1500-3000KU / kg aflatoxin degrading enzyme, 1-10g / kg 2-deoxy-D-glucose;

[0007] The carrier replenishes the surplus.

[0008] The auxiliary agent provided in the present application is mixed in the feed and fed together for fish consumption. Under normal conditions, the feed enters the body of the fish, and the starch is converted into sugar under the action of intestinal blood amylase. The auxiliary agent provided in the present application inhibits glycolysis by acting on hexokinase, the rate-limiting step of glycolysis, so that this part of the inhibited sugar cannot be converted into pyruvate, thereby achieving the generation of acetyl coenzyme A, and then cannot be converted into fat, reducing the formation of fatty liver, hypertrophy, whitening or other liver lesions in fish. And this part of the inhibited sugar can be used normally by the fish, such as for providing energy or converting into blood sugar, ensuring the normal growth of the fish and providing it with sufficient energy.

[0009] The combination of chromium picolinate and 2-deoxy-D-glucose can lower blood sugar, promote the entry of blood sugar into cells, reduce the burden on the liver to process glucose into fat, and further promote the effect of 2-deoxy-D-glucose.

[0010] In addition, the auxiliary agent of the present application also has the effects of protecting the liver, optimizing intestinal bacteria, promoting fat absorption, anti-inflammatory, strong anti-oxidation, and degrading aflatoxin, further ensuring liver function.

[0011] Preferably, vitamin B 12 It is 5000-6000ug / kg, vitamin C is 500-600mg / kg, astaxanthin is 500-550mg / kg, taurine is 20000-25000mg / kg, chromium picolinate (calculated as chromium) is 150-180mg / kg, aflatoxin degrading enzyme is 1800-2300KU / kg, and 2-deoxy-D-glucose is 3-8g / kg.

[0012] In actual production, if the dosage of 2-deoxy-D-glucose is too small, it will not have a good blocking effect, and fatty liver, liver hypertrophy and other conditions of carnivorous fish will not be effectively alleviated.

[0013] By adopting the above technical solution, the amount of 2-deoxy-D-glucose in the additive is further limited, so that carnivorous fish can generate an appropriate amount of fat without forming fatty liver after eating feed containing the additive.

[0014] Preferably, the carrier is a mixture of one or more of rice bran, stone powder, attapulgite clay, soybean meal, and rice husk powder.

[0015] Preferably, the carrier is rice bran, stone powder and attapulgite clay, the stone powder is 1500-10000 mg / kg, the attapulgite clay is 10-50 g / kg, and the rice bran makes up the balance.

[0016] If the raw materials in the auxiliary agent cannot be fully mixed, it is easy for the fish to eat the auxiliary agent, but the content of effective ingredients in the auxiliary agent is low, thus failing to achieve a good effect in improving fatty liver.

[0017] By adopting the above technical scheme, the type and proportion of the carrier are further limited. Under this condition, the particle size of the carrier is closer to the other raw materials of the auxiliary agent, so that it has a more sufficient mixing and breaking effect. The auxiliary agent does not absorb moisture, does not clump, and is not easy to generate static electricity, thereby playing a good role in improving fish fatty liver.

[0018] Preferably, when the carrier comprises attapulgite clay, the attapulgite clay is modified by the following steps:

[0019] Mixing attapulgite clay with water to prepare an attapulgite clay suspension;

[0020] Dissolve 2-deoxy-D-glucose and chitosan in water, mix them evenly at 300-500W and 1200-1500r / min to obtain a solution for use;

[0021] The standby liquid and the attapulgite clay suspension are mixed until uniform, allowed to stand, dried, and ground to obtain modified attapulgite clay.

[0022] When feed and additives are mixed and granulated, a feed pellet machine is needed. During the operation of the feed pellet machine, static electricity will be generated, which will cause over-dry or over-fine active ingredients to be adsorbed onto the equipment, resulting in the loss of additives and inability to mix with the feed.

[0023] By adopting the above technical scheme, the attapulgite clay is modified by special means, and 2-deoxy-D-glucose is mixed with it in advance, so that the 2-deoxy-D-glucose is firmly adsorbed on the attapulgite clay, forming a whole with the attapulgite clay, the particles become larger, are not easily affected by static electricity, and can play a more full role in the fish body.

[0024] In addition, 2-deoxy-D-glucose and chitin are ultrasonically dispersed and mixed at a specific power and speed in advance, and the fibrous structure of chitin will network 2-deoxy-D-glucose, so that 2-deoxy-D-glucose is not easy to fall off. Then the chitin attached with 2-deoxy-D-glucose reacts with the attapulgite clay suspension. The surface of attapulgite clay has a negative charge, and chitin has a positive charge. After the two come into contact, they are quickly attracted and 2-deoxy-D-glucose is firmly fixed and adsorbed to form a special structure. The modified attapulgite clay is non-charged and not easily affected by static electricity; 2-deoxy-D-glucose is firmly adsorbed on the surface of attapulgite clay through multiple means, and is not easy to fall off, which is convenient for its subsequent efficacy in the fish body.

[0025] Preferably, the mass ratio of 2-deoxy-D-glucose to chitin is 1:(1-3).

[0026] By adopting the above technical solution, the mass ratio of 2-deoxy-D-glucose to chitin is further limited, thereby ensuring that 2-deoxy-D-glucose has sufficient attachment sites on chitin.

[0027] In a second aspect, the present application provides a method for preparing an adjuvant for treating or preventing fatty liver of carnivorous fish, using the following technical solution:

[0028] A method for preparing an adjuvant for treating or preventing fatty liver of carnivorous fish comprises the following steps:

[0029] Mix all the ingredients together and stir until smooth to get the finished product.

[0030] By adopting the above technical solution, the finished product can be obtained after mixing various raw materials. The preparation steps are simple and convenient, which is more convenient for staff to implement and is suitable for promotion in this field.

[0031] Preferably, when mixing the raw materials, 50-70% of the carrier and vitamin B 12 , vitamin C, astaxanthin, taurine, chromium picolinate, aflatoxin degrading enzyme, and 2-deoxy-D-glucose are mixed until uniform, and then the remaining carrier is added and mixed again to obtain the finished product.

[0032] By adopting the above technical solution, the carrier is added in two batches, which is conducive to more complete mixing of the remaining raw materials and the carrier, thereby reducing the occurrence of agglomeration.

[0033] In summary, this application has the following beneficial effects:

[0034] 1. The auxiliary agent provided in the present application inhibits glycolysis by acting on hexokinase, the rate-limiting step of glycolysis, so that this part of the inhibited sugar cannot be converted into pyruvate, thereby achieving the generation of acetyl coenzyme A, and then cannot be converted into fat and cannot be oxidized, thereby reducing the formation of fatty liver in fish.

[0035] 2. The present application adopts special means to modify the attapulgite clay in the auxiliary agent, and mixes 2-deoxy-D-glucose therewith in advance, so that the 2-deoxy-D-glucose is firmly adsorbed on the attapulgite clay and forms a whole with the attapulgite clay. The particles become larger and are not easily affected by static electricity, so that they can play a more effective role in the fish body. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a picture of part of the fish liver in Experiment 2. The glass dish at the top is filled with the liver of the carnivorous fish that was fed with the adjuvant containing Example 1, and the glass dish at the bottom is filled with the liver of the carnivorous fish that was used as a blank control. DETAILED DESCRIPTION

[0037] The present application is further described in detail below with reference to the accompanying drawings and embodiments.

[0038] The raw materials used in the following examples and comparative examples are all commercially available products.

[0039] Example

[0040] Example 1

[0041] The total weight of the additives in the embodiment of the present application is 1 kg.

[0042] An adjuvant for treating or preventing fatty liver in carnivorous fish, comprising vitamin B 12 , vitamin C, astaxanthin, taurine, chromium picolinate (calculated as chromium), aflatoxin degrading enzyme, 2-deoxy-D-glucose, carrier.

[0043] The carriers are rice bran, stone powder and modified attapulgite clay.

[0044] The specific amounts of various raw materials are shown in Table 1.

[0045] Attapulgite clay is modified by the following steps:

[0046] Step a): mixing attapulgite clay and water in a ratio of 1:10 to prepare an attapulgite clay suspension.

[0047] Step b): dissolving 2-deoxy-D-glucose and chitosan in water, mixing and stirring for 5 minutes at 400W and 1300r / min to obtain a solution for use.

[0048] The mass ratio of 2-deoxy-D-glucose to chitin is 1:2, that is, the amount of chitin used is 13 g.

[0049] The amount of water used in step b) is 3 times the total mass of 2-deoxy-D-glucose and chitin.

[0050] Step c): placing the attapulgite clay suspension in a magnetic stirrer and stirring, then adding the standby liquid into the attapulgite clay suspension and mixing until uniform; and then switching to a constant temperature oscillator B for uniform oscillation.

[0051] The mixture is allowed to stand, dried and ground to obtain modified attapulgite clay.

[0052] The present application also discloses a method for preparing an adjuvant for treating or preventing fatty liver of carnivorous fish, comprising the following steps:

[0053] Step 1): Dry, crush and sieve various raw materials until the particle size reaches the required level.

[0054] Step 2): Evenly mix rice bran, stone powder and modified attapulgite clay to obtain a mixed carrier.

[0055] Add 60% of the mixed carrier into the waist drum mixer, and then continue to add vitamin B 12 , vitamin C, astaxanthin, taurine, chromium picolinate (calculated as chromium), aflatoxin degrading enzyme, and 2-deoxy-D-glucose are mixed therewith.

[0056] Step 3): Continue to add the remaining mixed carrier into the waist-drum mixer, continue to stir and mix, and obtain the auxiliary agent after being evenly mixed.

[0057] Example 2

[0058] An adjuvant for treating or preventing fatty liver of carnivorous fish, which is different from Example 1 in that:

[0059] The dosage of each raw material is different, please see Table 1 for details.

[0060] A method for preparing an adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that, in step b), mixing and stirring are performed under the conditions of 300W and 1500r / min; the mass ratio of 2-deoxy-D-glucose to chitin is 1:3, that is, the amount of chitin used is 3g.

[0061] In step 2), 50% of the mixed carrier is first mixed with the raw material.

[0062] Example 3

[0063] An adjuvant for treating or preventing fatty liver of carnivorous fish, which is different from Example 1 in that:

[0064] The dosage of each raw material is different, please see Table 1 for details.

[0065] A method for preparing an adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that, in step b), mixing and stirring are performed under the conditions of 500W and 1200r / min; the mass ratio of 2-deoxy-D-glucose to chitin is 1:1, that is, the amount of chitin used is 10g.

[0066] In step 2), 70% of the mixed carrier is first mixed with the raw material.

[0067] Example 4

[0068] An adjuvant for treating or preventing fatty liver of carnivorous fish, which is different from Example 1 in that:

[0069] The dosage of each raw material is different, please see Table 1 for details.

[0070] A method for preparing an auxiliary agent for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that the amount of chitosan used is 6 g.

[0071] Example 5

[0072] An adjuvant for treating or preventing fatty liver of carnivorous fish, which is different from Example 1 in that:

[0073] The dosage of each raw material is different, please see Table 1 for details.

[0074] A method for preparing an auxiliary agent for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that the amount of chitosan used is 16 g.

[0075]

[0076]

[0077] Example 6

[0078] An adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that the carrier is rice bran, stone powder, and modified attapulgite clay, the attapulgite clay is 3000 mg, the rice bran is 30 g, and the stone powder is supplemented to a balance to a total weight of 1 kg for the adjuvant.

[0079] Example 7

[0080] An adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that the carrier is unmodified attapulgite clay (commercially available), the stone powder is 0 mg, the modified attapulgite clay is 0 g, the rice bran is 0 g, and the commercially available attapulgite clay is used to supplement the total mass of the adjuvant to 1 kg.

[0081] Example 8

[0082] An adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that, in step b), mixing is performed under the conditions of 50W and 100r / min.

[0083] Example 9

[0084] An adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that, in step b), the mass ratio of 2-deoxy-D-glucose to chitin is 1:0.5, that is, the amount of chitin used is 3.25 g.

[0085] Example 10

[0086] An adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that, in step b), the mass ratio of 2-deoxy-D-glucose to chitin is 1:5, that is, the amount of chitin used is 32.5 g.

[0087] Embodiment 11

[0088] An adjuvant for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that in step b), chitosan is replaced by carboxymethyl cellulose, that is, the amount of chitosan is 0 g and the amount of carboxymethyl cellulose is 13 g.

[0089] Comparative Example

[0090] Comparative Example 1

[0091] An auxiliary agent for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that 2-deoxy-D-glucose is replaced by rice bran of equal mass, the amount of 2-deoxy-D-glucose used is 0 g, and the rice bran is adjusted to add to a total weight of 1 kg of the auxiliary agent.

[0092] Comparative Example 2

[0093] An auxiliary agent for treating or preventing fatty liver in carnivorous fish, which is different from Example 1 in that the amount of 2-deoxy-D-glucose used is 0.5 g, and rice bran is adjusted to supplement the total weight of the auxiliary agent to 1 kg.

[0094] Performance test test sample: the adjuvant and the feed are mixed in a ratio of 1:1000 to obtain a compound feed. The adjuvant is from Example 1-11 and Comparative Example 1-2.

[0095] Blank control sample: feed without additives.

[0096] Test 1-Physical and Chemical Testing

[0097] Select a number of healthy California bass juveniles with no body damage, place them in an aquarium for 5 days to stabilize them, and stop feeding them for 24 hours. Then select juveniles with an initial body weight of 9.7±0.2g, divide them into 15 groups, and feed them with the corresponding test samples for 30 days. Each group has 3 replicates, and each replicate has 30 fish.

[0098] Each group of experimental fish was fed twice a day (8 am and 16 pm), and the amount of feeding was enough to make the fish full. The water was changed once a day, 50% each time. Oxygen was provided 24 hours a day, and salinity and water temperature were monitored regularly.

[0099] After feeding on the last afternoon of the experiment, the fish were counted, weighed, and sampled the next morning. Twelve fish were randomly selected from each replicate to measure the corresponding indicators.

[0100] Indicator calculation formula:

[0101] Obesity (%) = 100*fish weight / fish length 3

[0102] Specific growth rate (%) = 100*(ln final weight - ln initial weight) / number of days of experiment

[0103] Liver to body ratio (%) = 100*liver mass / body weight

[0104] Viscera to body ratio (%) = 100*viscera mass / body weight

[0105] The results are mean ± standard error.

[0106] The results of Experiment 1 are detailed in Table 2.

[0107] Table 2

[0108]

[0109]

[0110] According to the comparison of the test data of Examples 1-3 with Comparative Examples 1-2 and the blank control in Table 2, it can be seen that the California bass that eat the feed containing the additives of Examples 1-3 have a higher specific growth rate, better obesity, lower liver-to-body ratio, and lower organ-to-body ratio than the California bass that eat Comparative Examples 1-2 and the blank control, indicating that the additives provided by the examples of the present application are used in feed, which can effectively improve the effect of starch on the liver of carnivorous fish, play a role in preventing and treating fatty liver, and the fish raised are also healthier. In addition, the final weight of Examples 1-3 is heavier than that of Comparative Examples 1-2 and the blank control, indicating that the additives provided by the present application help to improve the growth performance of carnivorous fish and provide sufficient energy.

[0111] According to the comparison of the test data of Example 1 with Examples 4 and 5 in Table 2, it can be seen that further limiting the amount of 2-deoxy-D-glucose in the adjuvant helps to improve the effect of preventing and treating fatty liver. For Example 5, the amount of 2-deoxy-D-glucose used is increased, but the relevant data is not as good as that of Example 4. The inventor speculates that it may be because although the amount of 2-deoxy-D-glucose is increased, the amount of attapulgite clay modified together is not increased, and the adsorption effect of part of 2-deoxy-D-glucose may be affected, thereby affecting the overall data. But overall, Examples 4 and 5 still maintain a good level of prevention and treatment.

[0112] According to the comparison of the test data of Example 1 with Examples 6 and 7 in Table 2, it can be seen that Example 1 is better than Examples 6 and 7 in promoting the growth of carnivorous fish and preventing fatty liver in carnivorous fish. This shows that the further selection of carrier types and the limitation of the dosage have a positive promoting effect on the effective substances such as 2-deoxy-D-glucose.

[0113] According to the comparison of the test data of Example 1 and Example 8 in Table 2, when modifying 2-deoxy-D-glucose and attapulgite clay, the mixing conditions of chitin and 2-deoxy-D-glucose cannot be changed at will. Only under the specific conditions specified in this application can 2-deoxy-D-glucose be more firmly adsorbed on attapulgite clay to the greatest extent.

[0114] According to the comparison of the test data of Example 1 and Example 11 in Table 2, chitosan can cooperate with 2-deoxy-D-glucose, and rashly replacing it with other substances will destroy the special cooperation effect. Combined with the comparison of the test data of Examples 9 and 10, it can be seen that too much or too little chitosan has a greater effect on the cooperation with 2-deoxy-D-glucose, so not only does chitosan need to cooperate with 2-deoxy-D-glucose, but the ratio of the two needs to be further limited.

[0115] Test 2 - Visual observation

[0116] Ten fish were randomly selected from each group of fish that had completed the test in Experiment 1, and the fish were dissected to observe the liver condition, score them according to the standard, and record the average score.

[0117] Standard: The score is from 0 to 5. The larger the score, the more serious the lesion. For example, 0 means no hypertrophy or other lesions, and 5 means severe hypertrophy or other lesions.

[0118] The results of Experiment 2 are detailed in Table 3.

[0119] Table 3

[0120] Category Fraction Example 1 0 Example 2 1.0 Example 3 0.9 Example 4 0.6 Example 5 0.5 Example 6 0.7 Example 7 1.2 Example 8 0.9 Example 9 0.4 Example 10 0.6 Embodiment 11 1.3 Comparative Example 1 3.0 Comparative Example 2 2.5 Blank control 3.3

[0121] Reference Figure 1 The glass dish at the top contained the liver of the carnivorous fish that had been fed with the additive in Example 1. The liver was ruddy in color without any whitish color or marbling, and the liver was of normal size without any obesity. The glass dish at the bottom contained the liver of the carnivorous fish that was used as the blank control. Most of them were whitish in color and had enlarged and swollen livers.

[0122] According to the test data of Examples 1-11 in Table 3, after carnivorous fish eat feed containing an additive prepared using a specific amount of 2-deoxy-D-glucose, they basically do not develop severe liver hypertrophy and other pathological changes, which is roughly consistent with the results of Experiment 1.

[0123] According to the comparison of the test data of Example 1 with Comparative Examples 1-2 and the blank control in Table 3, it can be seen that the carnivorous fish of Comparative Examples 1, 2 and the blank control have more serious pathological changes such as fish liver hypertrophy, and the effect is far inferior to that of Example 1.

[0124] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. An adjuvant for treating or preventing fatty liver of carnivorous fish, characterized in that: Including the following ingredients: 5000-7000 ug / kg Vitamin B 12 , 500-750mg / kg vitamin C, 500-600mg / kg astaxanthin, 20000-30000mg / kg taurine, 150-200mg / kg chromium picolinate (calculated as chromium), 1500-3000KU / kg aflatoxin degrading enzyme, 1-10g / kg 2-deoxy-D-glucose; The carrier replenishes the surplus.

2. The adjuvant for treating or preventing fatty liver of carnivorous fish according to claim 1, characterized in that: Vitamin B 12 It is 5000-6000 ug / kg, vitamin C is 500-600mg / kg, astaxanthin is 500-550mg / kg, taurine is 20000-25000mg / kg, chromium picolinate (calculated as chromium) is 150-180mg / kg, aflatoxin degrading enzyme is 1800-2300KU / kg, and 2-deoxy-D-glucose is 3-8g / kg.

3. The adjuvant for treating or preventing fatty liver of carnivorous fish according to claim 1, characterized in that: The carrier is a mixture of one or more of rice bran, stone powder, attapulgite clay, soybean meal and rice husk powder.

4. The adjuvant for treating or preventing fatty liver of carnivorous fish according to claim 3, characterized in that: The carriers are rice bran, stone powder and attapulgite clay, wherein the stone powder is 1500-10000 mg / kg, the attapulgite clay is 10-50 g / kg, and the rice bran is used to supplement the balance.

5. A method for preparing an adjuvant for treating or preventing fatty liver of carnivorous fish according to any one of claims 1 to 4, characterized in that: The following steps are involved: Mix all the ingredients together and stir until smooth to get the finished product.

6. The method for preparing the auxiliary agent for treating or preventing fatty liver of carnivorous fish according to claim 5, characterized in that: When mixing the raw materials, first take 50-70% of the carrier and vitamin B 12 , vitamin C, astaxanthin, taurine, chromium picolinate, aflatoxin degrading enzyme, and 2-deoxy-D-glucose are mixed until uniform, and then the remaining carrier is added and mixed again to obtain the finished product.

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