Preparation method and use method of traditional Chinese medicine feed additive for culturing carps in carbonate water area

Abstract

The invention discloses a preparation method and a use method of a traditional Chinese medicine feed additive for culturing carps in a carbonate water area, and relates to a preparation method and a use method of a traditional Chinese medicine feed additive for culturing carps. The invention aims to solve the technical problems of slow growth, immune stress and even death of cultured carps caused by carbonate in a saline-alkali water area at present. According to the invention, the hedyotis diffusa / sculellaria barbata drug pair is added into the feed, so that the damage of carbonate stress to the immune system of the carp can be relieved by adjusting the activity of tissue immune related enzymes and inducing cell autophagy, and the immunocompetence of the carp in a carbonate water body is improved; the oldenlandia diffusa / sculellaria barbata drug pair is beneficial to relieving oxidative stress of the carps in the carbonate water area and removing damaged organelles, and finally is beneficial to improving the survival rate of the carps in the carbonate water area and enhancing the adaptive capacity of the carps to the saline-alkali environment.

Description

Technical Field

[0001] The present invention relates to a preparation method and a usage method of a traditional Chinese medicine feed additive for culturing carp. Background Art

[0002] Saline-alkali waters refer to waters where the content of salts and alkaline substances in the water body is significantly higher than that of normal fresh water due to natural or human factors. Such waters are widely distributed around the world, especially more common in arid and semi-arid regions, coastal areas, and inland closed drainage areas. Research shows that when fish enter waters with high alkalinity, HCO 3- and CO3 2- ions can cause damage to the gills and skin surface of fish, disrupt the ion exchange of the body, affect the acid-base balance of fish, and thus affect the survival of fish.

[0003] Currently, the research on culturing carp in saline-alkali waters mainly focuses on the following aspects: breeding of salt-tolerant and alkali-tolerant varieties, water quality regulation, and nutritional regulation. However, the breeding cycle of salt-tolerant and alkali-tolerant varieties is long, the cost is high, and the price is expensive, making it difficult to apply in production. The water quality regulation technology is usually complex in operation and the price of the medicament is high, making it difficult to promote and use on a large scale. The stress resistance and immunity of fish can be enhanced by adjusting the feed formula, and the survival rate of fish can be improved. As a natural, green, and safe additive, traditional Chinese medicine has the effects of anti-stress and enhancing immunity. Some traditional Chinese medicines are low in price and easy to prepare, which well solves the problem of being difficult to apply in production. Summary of the Invention

[0004] The present invention aims to solve the technical problem that carbonates in current saline-alkali waters cause slow growth, immune stress, and even death of cultured carp, and provides a preparation method and a usage method of a traditional Chinese medicine feed additive for culturing carp in carbonate waters.

[0005] The preparation method of the traditional Chinese medicine feed additive for culturing carp in carbonate waters of the present invention is carried out according to the following steps:

[0006] First, dry whole herbs of Hedyotis diffusa and Scutellaria barbata are mixed together, distilled water is added, after soaking for 2 h to 2.5 h, it is placed on a heating plate, boiled at 400 °C to 410 °C, then the heating plate is cooled to 320 °C to 330 °C to keep slightly boiling for 45 min to 50 min, and then naturally cooled to room temperature. The liquid is filtered out and the medicinal liquid and medicinal residues are respectively retained;

[0007] The mass ratio of the dry whole herb of Scutellaria barbata to the dry whole herb of Hedyotis diffusa is 2:(1 - 1.2);

[0008] The mass ratio of the dry whole herb of Scutellaria barbata to the volume of distilled water is 1 g:(25 mL - 30 mL);

[0009] II. Add distilled water with the same volume as that in Step 1 to the medicinal residues obtained at the end of Step 1, boil it at 400°C to 410°C until boiling, then lower the heating plate temperature to 320°C to 330°C and keep it boiling gently for 45 min to 50 min, and then let it cool naturally to room temperature. Filter out the liquid and retain the medicinal liquid, and mix the two portions of medicinal liquid obtained in Step 1 and Step 2 together;

[0010] III. Place the mixed medicinal liquid in Step 2 in a rotary evaporator and evaporate it under rotation at 80°C to 85°C until the concentration of the crude drug amount in the medicinal liquid reaches 1 g / mL to 1.5 g / mL, obtaining a traditional Chinese medicine feed additive.

[0011] The concentration of the crude drug amount in Step III is the sum of the masses of the dried whole herb of Hedyotis diffusa and the dried whole herb of Scutellaria barbata in Step I divided by the volume of the medicinal liquid after rotary evaporation in Step III.

[0012] The usage method of the traditional Chinese medicine feed additive of the present invention applied to breeding carp in carbonate water areas is carried out according to the following steps:

[0013] The first case: Add the above-prepared traditional Chinese medicine feed additive during the preparation of the basic feed: Add the above-prepared traditional Chinese medicine feed additive to the basic feed before granulating the basic feed. Each 1 kg of the basic feed contains 20 g to 22 g of the crude drug amount, and then proceed with normal feed granulation;

[0014] The second case: Add the above-prepared traditional Chinese medicine feed additive to the finished basic feed: Add distilled water to the above-prepared traditional Chinese medicine feed additive for dilution. The volume ratio of the above-prepared traditional Chinese medicine feed additive to the added distilled water is 2:(3 - 3.5); then add sodium alginate for mixing, and the mass of sodium alginate is 1% of the mass of the diluted traditional Chinese medicine feed additive; then mix it with the finished basic feed until there is no adhesion between the particles of the basic feed, spread it out flat, dry it in a cool and dry condition, and store it in a 4°C refrigerator for later use; each 1 kg of the finished basic feed contains 20 g to 22 g of the crude drug amount.

[0015] The object of the present invention is to alleviate the problem that carbonates in saline-alkali waters cause slow growth, immune stress and even death of cultured carp. In the present invention, the herb pair of *Hedyotis diffusa* / *Scutellaria barbata* is added to the basic feed. As a traditional Chinese medicine formula, it has extremely high medicinal value. Both are rich in chemical substances such as active polysaccharides, alkaloids, and polyphenols, and have pharmacological effects such as clearing heat and detoxifying, reducing swelling and dissipating stasis, and eliminating various inflammations. Components such as polysaccharides contained in *Hedyotis diffusa* can stimulate the activities of immune cells, such as macrophages and T lymphocytes, enhance the phagocytic function of macrophages, and can also regulate the secretion of cytokines, thereby enhancing the body's immune defense. Substances such as flavonoids in *Scutellaria barbata* can improve the functional state of immune organs, regulate the signal pathways of immune cells, and regulate immune responses. The ability to scavenge OH and protect DNA is mainly contributed by *Scutellaria barbata*, while the chelating ability is mainly related to the proportion of *Hedyotis diffusa*. The two work together to regulate the body's immune ability in many aspects. To sum up, the herb pair of *Hedyotis diffusa* / *Scutellaria barbata* can relieve oxidative stress and decreased immunity induced by carbonate stress by regulating the activities of tissue immunity-related enzymes, scavenging free radicals, antioxidation and inducing autophagy, which is beneficial to improving the survival rate of carp in carbonate waters, enhancing the immune ability of carp in carbonate water bodies, and strengthening the adaptability of carp to saline-alkali environments. Description of the Drawings

[0016] Figure 1 It is a data graph of the effect of the lotus-snake medicine pair on the content of catalase in the liver tissue of carp in Experiment 3;

[0017] Figure 2 It is a data graph of the effect of the lotus-snake medicine pair on the content of malondialdehyde in the liver tissue of carp in Experiment 3;

[0018] Figure 3 It is a data graph of the effect of the lotus-snake medicine pair on the content of superoxide dismutase in the liver tissue of carp in Experiment 3;

[0019] Figure 4 It is a data graph of the effect of ACP in Experiment 3;

[0020] Figure 5 It is a data graph of the effect of AKP in Experiment 3;

[0021] Figure 6 It is a data graph of the effect on the expression level of the immune gene TNF-α of carp in Experiment 4;

[0022] Figure 7 It is a data graph of the effect on the expression level of the immune gene IL-1β of carp in Experiment 4;

[0023] Figure 8 It is a data graph of the effect on the expression level of the immune gene IL-8 of carp in Experiment 4;

[0024] Figure 9Data graph showing the effect on the expression level of carp SOD antioxidant gene in Experiment 4;

[0025] Figure 10 Data graph showing the effect on the expression level of carp CAT antioxidant gene in Experiment 4. Specific implementation manners

[0026] Specific implementation manner 1: This implementation manner is a preparation method of a traditional Chinese medicine feed additive for culturing carp in carbonate water area, and specifically is carried out according to the following steps:

[0027] 1. Mix the dry whole herb of Hedyotis diffusa and the dry whole herb of Scutellaria barbata, add distilled water, soak for 2 h - 2.5 h, then place on a heating plate, boil at 400 °C - 410 °C, then reduce the temperature of the heating plate to 320 °C - 330 °C and keep slightly boiling for 45 min - 50 min, and then naturally cool to room temperature. Filter out the liquid and retain the medicinal liquid and the medicinal residues respectively;

[0028] The mass ratio of the dry whole herb of Scutellaria barbata to the dry whole herb of Hedyotis diffusa is 2:(1 - 1.2);

[0029] The mass ratio of the dry whole herb of Scutellaria barbata to the volume of distilled water added is 1 g:(25 mL - 30 mL);

[0030] 2. Add the same volume of distilled water as in step 1 to the medicinal residues obtained at the end of step 1, boil at 400 °C - 410 °C, then reduce the temperature of the heating plate to 320 °C - 330 °C and keep slightly boiling for 45 min - 50 min, and then naturally cool to room temperature. Filter out the liquid and retain the medicinal liquid, and mix the two portions of medicinal liquid obtained in step 1 and step 2 together;

[0031] 3. Place the mixed medicinal liquid in step 2 in a rotary evaporator, rotate and evaporate at 80 °C - 85 °C until the concentration of the medicinal liquid is concentrated to the concentration of crude drug amount of 1 g / mL - 1.5 g / mL, and obtain the traditional Chinese medicine feed additive.

[0032] Specific implementation manner 2: The difference between this implementation manner and specific implementation manner 1 is that: the mass ratio of the dry whole herb of Hedyotis diffusa to the dry whole herb of Scutellaria barbata in step 1 is 1:2. Others are the same as specific implementation manner 1.

[0033] Specific implementation manner 3: The difference between this implementation manner and specific implementation manner 1 or 2 is that: in step 1, boil at 400 °C, then reduce the temperature of the heating plate to 320 °C and keep slightly boiling for 45 min. Others are the same as specific implementation manner 1 or 2.

[0034] Embodiment 4: The difference between this embodiment and any one of Embodiments 1 to 3 is that: in Step 3, rotary evaporation is carried out in a water bath at a temperature of 80 °C. Others are the same as any one of Embodiments 1 to 3.

[0035] Embodiment 5: The difference between this embodiment and Embodiment 4 is that: the rotation speed during rotary evaporation in Step 3 is 80 r / min. Others are the same as Embodiment 4.

[0036] Embodiment 6: The difference between this embodiment and Embodiment 5 is that: in Step 3, the medicinal liquid is concentrated until the concentration of the crude drug amount is 1 g / mL to obtain the traditional Chinese medicine feed additive. Others are the same as Embodiment 5.

[0037] Embodiment 7: This embodiment is the usage method of the traditional Chinese medicine feed additive applied to cultivating carp in carbonate water area in Embodiment 1, specifically:

[0038] The first case, adding the prepared traditional Chinese medicine feed additive during the preparation process of the basic feed: adding the prepared traditional Chinese medicine feed additive into the basic feed before granulation of the basic feed, with 20 g to 22 g of crude drug amount contained in every 1 kg of the basic feed, and then normal feed granulation can be carried out;

[0039] The second case, adding the prepared traditional Chinese medicine feed additive to the finished basic feed: adding distilled water to the prepared traditional Chinese medicine feed additive for dilution, and the volume ratio of the prepared traditional Chinese medicine feed additive to the added distilled water is 2:(3 - 3.5); then adding sodium alginate for mixing, and the mass of sodium alginate is 1% of the mass of the diluted traditional Chinese medicine feed additive; then mixing with the finished basic feed until there is no adhesion between the particles of the basic feed, spreading out, drying in a cool and dry condition, and storing in a bag in a 4 °C refrigerator for standby; 20 g to 22 g of crude drug amount is contained in every 1 kg of the finished basic feed.

[0040] Embodiment 8: The difference between this embodiment and Embodiment 7 is that: in the second case, the mixing method with the finished basic feed is manual grasping and mixing. Others are the same as Embodiment 7.

[0041] The present invention is verified by the following tests:

[0042] Test 1: This test is a preparation method of a traditional Chinese medicine feed additive applied to cultivating carp in carbonate water area, and is specifically carried out according to the following steps:

[0043] 1. Mix 10 g of dried whole herb of *Hedyotis diffusa* and 20 g of dried whole herb of *Scutellaria barbata* together, add 500 mL of distilled water, soak for 2 h, then place on a heating plate, boil at 400 °C, then lower the temperature of the heating plate to 320 °C and keep it boiling gently for 45 min, and then let it cool naturally to room temperature. Filter out the liquid and retain the medicinal liquid and the residue separately;

[0044] 2. Add another 500 mL of distilled water to the residue obtained in the last step of step 1, boil at 400 °C, then lower the temperature of the heating plate to 320 °C and keep it boiling gently for 45 min, and then let it cool naturally to room temperature. Filter out the liquid and retain the medicinal liquid. Mix the two portions of medicinal liquid obtained in step 1 and step 2 together;

[0045] 3. Place the mixed medicinal liquid obtained in step 2 in a rotary evaporator, evaporate it by rotation in a water bath at 80 °C with a rotation speed of 80 r / min, and concentrate the medicinal liquid to a concentration of 1 g / mL of the crude drug amount to obtain a traditional Chinese medicine feed additive, denoted as the *Scutellaria barbata-Hedyotis diffusa* pair additive;

[0046] The concentration of the crude drug amount in step 3 is the sum of the masses of the dried whole herb of *Hedyotis diffusa* and the dried whole herb of *Scutellaria barbata* in step 1 (30 g) divided by the volume of the medicinal liquid after rotary evaporation in step 3 (30 mL);

[0047] 4. Add the above-prepared traditional Chinese medicine feed additive to the finished basic feed: Dilute the above-prepared traditional Chinese medicine feed additive with distilled water. The volume ratio of the above-prepared traditional Chinese medicine feed additive to the added distilled water is 2:3; then add sodium alginate and mix. The mass of sodium alginate is 1% of the mass of the diluted traditional Chinese medicine feed additive; then manually mix it with the finished basic feed by stirring until there is no adhesion between the particles of the basic feed, spread it out flat, dry it in a cool and dry place, and store it in a 4 °C refrigerator for later use to obtain the traditional Chinese medicine feed; Each 1 kg of the finished basic feed contains 20 g of the crude drug amount; The finished basic feed is purchased from the fish-use extruded compound feed - Fish Seed Health No. 6 product of Huai'an Tongwei Feed Co., Ltd. and is in granular form.

[0048] Experiment 2: Effect of the *Scutellaria barbata-Hedyotis diffusa* pair additive on the immune ability of carp:

[0049] Two weeks of acclimation was carried out before the stress test. During the acclimation period, the fish were fed three times a day (at 9:00, 13:00, and 16:00). The water quality was the aquaculture water from the Aquaculture Laboratory of Jilin Agricultural University, with a water temperature of 20.3 ± 0.5 °C, a pH of 7.37 ± 0.2, and a dissolved oxygen of ≥ 7 mg / L. After the acclimation, 180 juvenile carps with healthy bodies and an initial mass of 3.4 ± 0.08 g were selected and randomly divided into three groups, including a control group (CK), a NaHCO3 stress group (CA), and a lotus snake medicine pair stress group (BSA). Each group had three buckets, with 20 fish in each bucket, and each group had three replicates. The fish were reared for 8 weeks.

[0050] The water quality of the control group (CK) was the same as that during the above acclimation period;

[0051] In the NaHCO3 stress group (CA), NaHCO3 was added to the water quality of the control group to a concentration of 50 mmol / L and a pH of 8.9. The control group (CK) and the NaHCO3 stress group (CA) were fed the same variety of basal feed, the fish-expanding compound feed - Fish Seed Health 6 product purchased from Huai'an Tongwei Feed Co., Ltd., which was granular;

[0052] The water quality of the lotus snake medicine pair stress group (BSA) was the same as that of the NaHCO3 stress group (CA), and the Chinese medicine feed prepared in Step 4 of Experiment 1 was fed.

[0053] For the above feeding, the feed was fed at 1% of the body weight of the experimental fish each time, and 80% of the water was changed every two days.

[0054] After 8 weeks of rearing, feeding was stopped for 24 h, and 10 carps were randomly taken from each bucket for weighing, which was used for the calculation of the growth performance of each group. The specific calculation formula for the growth performance is shown in Table 1 below.

[0055] Table 1

[0056] Index Formula Weight gain rate (%) <![CDATA[(W t / W0)×100%]]> Specific growth rate (% / d) <![CDATA[(lnW t -lnW0) / t×100%]]>

[0057] Among them, W0 is the mass of the fish at the beginning of the experiment, in g; W t is the mass of the fish at the end of the experiment, in g; t is the experimental time, in d.

[0058] The data of the growth performance of juvenile carps in Experiment 2 are shown in Table 2 below.

[0059] Table 2

[0060]

[0061] The above results show that: saline-alkali stress significantly reduced the survival rate of carps, while the Chinese medicine feed of the lotus snake medicine pair could significantly improve the growth performance of carps and the weight gain rate of carps under saline-alkali stress.

[0062] Experiment 3: After the feeding experiment in Experiment 2 ended, the fish were starved for 24 hours. Then, 15 fish were randomly selected from each group, anesthetized, and their liver tissues were collected on an ice plate, quickly frozen in liquid nitrogen, and stored at -80 °C for the determination of immune enzyme and antioxidant enzyme activities.

[0063] The antioxidant capacity was determined using a kit to measure antioxidant indices in the liver of carp, including superoxide dismutase (SOD, WST-1 method, U / mg), malondialdehyde (MDA, TBA method, nmol / g), and catalase (CAT, ammonium molybdate method, U / mg).

[0064] Figure 1 It is a data graph showing the effect of lotus snake medicine pair on the content of catalase in the liver tissue of carp in Experiment 3. Figure 2 It is a data graph showing the effect of lotus snake medicine pair on the content of malondialdehyde in the liver tissue of carp in Experiment 3. Figure 3 It is a data graph showing the effect of lotus snake medicine pair on the content of superoxide dismutase in the liver tissue of carp in Experiment 3.

[0065] Figures 1 - 3 The results show that during the stress period, compared with the CK group, the activities of SOD and CAT in the liver of carp in the CA group decreased significantly (P < 0.05), and the activity of MDA increased significantly (P < 0.05), causing oxidative stress in carp. Compared with the CA group, the activities of SOD and CAT in the liver of carp in the BSA group increased significantly (P < 0.05), and the activity of MDA decreased significantly (P < 0.05), enhancing the antioxidant capacity of carp.

[0066] Note: Excessive accumulation of MDA will lead to oxidative stress.

[0067] The immune indices in the liver of carp were measured: ACP, acid phosphatase (ELISA method, U / g), and AKP, alkaline phosphatase (microplate method, U / g).

[0068] Figure 4 It is a data graph showing the effect of ACP in Experiment 3. Figure 5 It is a data graph showing the effect of AKP in Experiment 3. The results show that during the stress period, compared with the CK group, the activities of ACP and AKP in the liver of carp in the CA group decreased significantly (P < 0.05), causing immune stress in carp. Compared with the CA group, the activities of ACP and AKP in the liver of carp in the BSA group increased significantly (P < 0.05), enhancing the immune function of carp.

[0069] Experiment 4: This experiment was to study the effect of lotus snake medicine pair on the expression levels of antioxidant and immune genes in carp, as follows:

[0070] This experiment adopted a single-factor factorial experimental design, which was divided into 3 groups: the control group (CK), the stress group (CA), and the BSA group. The specific breeding process was the same as that in Experiment 2. After the feeding experiment ended, the fish were starved for 24 h. Five fish were randomly selected from each group, anesthetized, and liver tissues were collected on an ice plate, quickly frozen in liquid nitrogen, and stored at -80 °C for the determination of the expression levels of immune and antioxidant genes. Two antioxidant-related genes were selected: SOD (gene sequence number: XM_026251256.1) and CAT (gene sequence number: XM_026203265.1). Three immune genes were selected: IL-1β (gene sequence number: AJ249137), IL-8 (gene sequence number: NC_039268.1), and TNF-α (gene sequence number: NC_039261.1). The genes were derived from the NCBI and Ensembl databases, and primers were designed using Primer3.

[0071] The data on the effects of the expression levels of immune-related genes in carp are as Figures 6 - 8 shown. During the stress period, compared with the CK group, the expression levels of TNF-α, IL-1β, and IL-8 genes in the livers of carp in the CA group increased significantly (P < 0.05). The results showed that NaHCO3 stress led to an increase in the expression levels of pro-inflammatory factors in the carp body and induced an inflammatory response in the carp body. Compared with the CA group, the expression levels of TNF-α, IL-1β, and IL-8 genes in the livers of carp in the BSA group decreased significantly (P < 0.05), effectively improving the inflammatory response in carp caused by NaHCO3 stress.

[0072] The data on the effects of the expression levels of antioxidant-related genes in carp are as Figures 9 - 10 shown. During the stress period, compared with the CK group, the expression levels of SOD and CAT genes in the livers of carp in the CA group decreased significantly (P < 0.05), causing oxidative damage to the body. Compared with the CA group, the expression levels of SOD and CAT genes in the livers of carp in the BSA group increased significantly (P < 0.05). The results showed that adding BSA to the basal diet effectively improved the oxidative stress response caused by NaHCO3 stress.

Claims

1. A preparation method of a traditional Chinese medicine feed additive for culturing carp in carbonate water areas, characterized in that The preparation method of the traditional Chinese medicine feed additive applied to culturing carp in carbonate water area is carried out according to the following steps: First, mix the dry whole herb of Oldenlandia diffusa and the dry whole herb of Scutellaria barbata, add distilled water, soak for 2h - 2.5h, then place on a heating plate, boil at 400℃ - 410℃, then lower the temperature of the heating plate to 320℃ - 330℃ and keep gently boiling for 45min - 50min, and then naturally cool to room temperature. Filter out the liquid and retain the medicinal liquid and the medicinal residues respectively; The mass ratio of the dry whole herb of Scutellaria barbata to the dry whole herb of Oldenlandia diffusa is 2:(1 - 1.2); The mass ratio of the dry whole herb of Scutellaria barbata to the volume of distilled water added is 1g:(25mL - 30mL); Second, add the same volume of distilled water as in step one to the medicinal residues obtained in the last step of step one, boil at 400℃ - 410℃, then lower the temperature of the heating plate to 320℃ - 330℃ and keep gently boiling for 45min - 50min, and then naturally cool to room temperature. Filter out the liquid and retain the medicinal liquid, and mix the two portions of medicinal liquid obtained in step one and step two together; Third, place the mixed medicinal liquid in step two in a rotary evaporator, rotate and evaporate at 80℃ - 85℃ until the concentration of the crude drug amount in the medicinal liquid is 1g / mL - 1.5g / mL, and obtain the traditional Chinese medicine feed additive.

2. The preparation method of a traditional Chinese medicine feed additive for culturing carp in carbonate water area according to claim 1, characterized in that The mass ratio of the dry whole herb of Oldenlandia diffusa to the dry whole herb of Scutellaria barbata described in step one is 1:

2.

3. The preparation method of a traditional Chinese medicine feed additive for culturing carp in carbonate water area according to claim 1, characterized in that In step one, boil at 400℃, then lower the temperature of the heating plate to 320℃ and keep gently boiling for 45min.

4. The preparation method of a traditional Chinese medicine feed additive for culturing carp in carbonate water areas according to claim 1, characterized in that In step three, rotate and evaporate in a water bath at 80℃.

5. The preparation method of a traditional Chinese medicine feed additive for culturing carp in carbonate water area according to claim 1, characterized in that The rotation speed during rotation evaporation in step three is 80r / min.

6. The preparation method of a traditional Chinese medicine feed additive for culturing carp in carbonate water area according to claim 1, characterized in that In step three, concentrate the medicinal liquid until the concentration of the crude drug amount is 1g / mL, and obtain the traditional Chinese medicine feed additive.

7. The usage method of a traditional Chinese medicine feed additive for culturing carp in carbonate water areas as described in claim 1, characterized in that The using method of the traditional Chinese medicine feed additive applied to culturing carp in carbonate water area is as follows: The first case: Add the above-prepared traditional Chinese medicine feed additive during the preparation of the basic feed: Add the above-prepared traditional Chinese medicine feed additive to the basic feed before granulating the basic feed. Each 1kg of the basic feed contains 20g - 22g of the crude drug amount, and then proceed with normal feed granulation; The second case: Add the above-prepared traditional Chinese medicine feed additive to the finished basic feed: Add distilled water to the above-prepared traditional Chinese medicine feed additive for dilution. The volume ratio of the above-prepared traditional Chinese medicine feed additive to the added distilled water is 2:(3 - 3.5); then add sodium alginate for mixing. The mass of sodium alginate is 1% of the mass of the diluted traditional Chinese medicine feed additive; then mix with the finished basic feed until there is no adhesion between the particles of the basic feed, lay it flat, dry it in a cool and dry condition, and store it in a 4℃ refrigerator for later use; each 1kg of the finished basic feed contains 20g - 22g of the crude drug amount.

8. The method for using a traditional Chinese medicine feed additive for culturing carp in carbonate water area according to claim 7, characterized in that In the second case, the mixing method with the finished basic feed is manual grasping and mixing.

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