Micropterus salmoides composite Chinese herbal medicine additive and preparation and periodic application method thereof

Abstract

The invention discloses a micropterus salmoides composite Chinese herbal medicine additive and a preparation and periodic application method thereof. The feed is prepared from radix scutellariae, dandelion, sappan wood and Chinese pulsatilla root according to a specific proportion through water decoction and concentration processes, so that the capability of resisting aeromonas veronii of the micropterus salmoides can be remarkably improved, the oxidation resistance and the immune performance are enhanced, and the feed is suitable for large-scale popularization and application. And the damage of high-dose herbal medicine to intestinal tracts is effectively relieved through an application mode of feeding the micropterus salmoides once every three days and continuously feeding the micropterus salmoides with the Chinese herbal medicine preparation for 28 days. Experiments show that the growth performance of micropterus salmoides can be remarkably improved by adding 0.1% of the feed, meanwhile, the feeding mode has no damage to livers, and the survival rate of fish bodies reaches 92% or above after toxin attacking of aeromonas veronii. The method has the advantages of low cost, high safety, easiness in industrialization and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of aquaculture, and particularly relates to a California sea bass composite Chinese herbal medicine additive and a preparation and periodic application method thereof. Background Art

[0002] While the intensive, high-density aquaculture of California bass (Micropterus salmoides) has increased production, it has also significantly increased the risk of Aeromonas spp. infection. Aeromonas secretes virulence factors such as hemolysins and enterotoxins, causing enteritis, ulcers, and sepsis, resulting in a mortality rate as high as 30%-50%, severely hindering the sustainable development of the industry. Traditional control strategies relying on antibiotics (such as enrofloxacin and florfenicol) are effective in the short term, but their long-term overuse exacerbates the spread of drug-resistant strains, disrupts the aquatic microbiome, and increases drug residue levels in aquatic products, threatening food safety and the environment.

[0003] Due to their multi-component and multi-target properties, Chinese herbal medicines have become a research hotspot for alternatives to antibiotics. Their active ingredients (such as berberine, chlorogenic acid, and polysaccharides) can achieve broad-spectrum antibacterial activity by disrupting pathogen biofilms and inhibiting virulence gene expression. They also activate fish's nonspecific immunity (such as the complement system and lysozyme activity), regulate intestinal microbiota homeostasis, and promote nutrient absorption and growth performance. However, the application of existing Chinese herbal additives faces significant bottlenecks. High doses or long-term feeding can easily cause intestinal morphological damage (such as villous atrophy and goblet cell reduction), leading to growth inhibition and even metabolic disorders. Summary of the Invention

[0004] In response to the difficulties in the existing technology, the present invention provides a composite Chinese herbal additive for California bass, and a preparation and periodic application method thereof. The intermittent feeding application mode (feeding California bass feed supplemented with Chinese herbal preparations once every three days for 28 days) effectively alleviates the damage to the intestines caused by high doses of herbal medicine. The Chinese herbal preparation is made by using Scutellaria baicalensis, Taraxacum mongolicum, Sappan wood, and Pulsatilla chinensis in a specific proportion and undergoing water decoction and concentration processes. It significantly improves the California bass's ability to resist Aeromonas vermiformis and enhances its antioxidant and immune properties.

[0005] One of the technical solutions of the present invention is to provide a method for preparing a California sea bass composite Chinese herbal medicine additive, comprising the following steps: (1) Soak the Chinese herbal medicines in 10 times the weight of water for 3 hours, then decoct for 40 minutes and filter out the medicinal liquid; add 30 times the weight of water to the filtered medicinal residues, decoct for another 40 minutes and filter out the medicinal liquid; the Chinese herbal medicines are scutellaria baicalensis, dandelion, sappan wood and pulsatilla chinensis; (3) Combine the two decoctions; (4) Concentrating the above medicinal solution to a crude drug content of 1 g / mL; (5) 20 parts by weight of Scutellaria baicalensis concentrated solution, 27 parts by weight of Taraxacum mongolicum concentrated solution, 25 parts by weight of Sappan wood concentrated solution, and 15 parts by weight of Pulsatilla chinensis concentrated solution were mixed to obtain a California sea bass compound Chinese herbal medicine additive.

[0006] The second technical solution of the present invention is to provide a California sea bass compound Chinese herbal medicine additive prepared by the above preparation method.

[0007] The third technical solution of the present invention is to provide a periodic application method of the above-mentioned California bass compound Chinese herbal additive, which is characterized in that the California bass compound feed is fed to the California bass once every three days for 28 days; the California bass compound feed contains 0.1wt% of the compound Chinese herbal additive; the compound Chinese herbal additive contains 20 weight parts of Scutellaria baicalensis concentrate, 27 weight parts of Taraxacum concentrate, 25 weight parts of Sappan wood concentrate and 15 weight parts of Pulsatilla scabra concentrate; the crude drug content in the concentrate is 1g / mL.

[0008] Furthermore, the preparation method of the California sea bass compound feed is: spraying the California sea bass compound Chinese herbal medicine additive at 0.1 wt % on the California sea bass basic feed.

[0009] The beneficial effects of the present invention are as follows: (1) by periodically intervening in the feeding of California sea bass, the use of a Chinese medicine compound preparation feed significantly reduced the superoxide dismutase (SOD), total antioxidant capacity (T-AOC) activity and malondialdehyde (MDA) concentration in the liver of California sea bass, while the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the serum of California sea bass showed a similar trend to the above indicators, thereby reducing the negative impact of 1.0 g / Kg HE in the feed on liver health.

[0010] (2) The feed containing the Chinese herbal compound preparation of the present invention has a simple preparation process and can be directly added to commercial feed without affecting the palatability of the feed. It is safe and non-toxic. The Chinese herbal raw materials used are low-priced, stable, and easy to industrialize.

[0011] (3) While reducing the workload of personnel, it can enhance the growth performance and disease resistance of fish. By feeding at intervals, it can reduce the damage to the intestine caused by high doses of herbal medicine, and has the dual effects of promoting growth and resisting disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 This is the feeding and experimental flow chart for the control and experimental groups; Figure 2 To study the effects of feeding compound Chinese herbal medicine (HE) at different intervals on the resistance of California seabass to Aeromonas vermiformis. Figure 3 The effects of regularly feeding compound Chinese herbal medicine every three days on the antioxidant and biochemical indices of California bass; Figure 4 Effects of herbal extracts on liver and intestinal morphology of California seabass. DETAILED DESCRIPTION

[0013] The following examples are used to further illustrate the present invention. Their purpose is to illustrate the present invention and should not be construed as limiting the scope of the present invention. Unless otherwise specified, all references are by weight and weight percentage.

[0014] Unless otherwise specified, the raw materials used in the present invention are conventional commercial products; the methods used in the present invention are conventional methods in the art unless otherwise specified.

[0015] The embodiments of the present invention are further described below with reference to a number of embodiments.

[0016] It should be clear that the embodiments described are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0017] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application. The singular forms "a", "an", "the" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms unless the context clearly indicates otherwise.

[0018] Example 1 (1) Soak scutellaria baicalensis, dandelion, sappan wood, and pulsatilla in water 10 times the weight of each drug for 3 hours, then decoct for 40 minutes and filter out the liquid; add 30 times the weight of water to the filtered residue, decoct for another 40 minutes, and filter out the liquid; (3) Combine the two decoctions; (4) Concentrating the above medicinal solution to a crude drug content of 1 g / mL; (5) 20 parts by weight of Scutellaria baicalensis concentrated solution, 27 parts by weight of Taraxacum mongolicum concentrated solution, 25 parts by weight of Sappan wood concentrated solution, and 15 parts by weight of Pulsatilla chinensis concentrated solution were mixed to obtain a California sea bass compound Chinese herbal medicine additive.

[0019] (6) Preparation of California bass feed: Take the California bass compound Chinese herbal medicine additive according to the dosage relationship and spray it on the California bass basic feed to prepare the California bass compound feed. The dosage of the Chinese herbal medicine compound preparation is 0.1% of the total weight of the California bass basic feed.

[0020] Example 2 The interval feeding experiment was conducted as follows: In the first stage, the juveniles were randomly divided into Con, HE101, HE102, HE103 and HE104 groups, with 25 fish in each tank and 3 replicates per group. Each fish tank contained 200 L of water, and the circulating water flow rate was 40 L·h -1 The water temperature was (24.0±2.0)°C, the pH was 7.5–8.0, and the dissolved oxygen was 5.0 mg·L -1 . The control group was fed a basal diet (without HE) daily, and all fish were fed at 8:30 am and 5:30 pm respectively until they were apparently full. Feeding was continued for 28 days. The HE101 group was initially fed a diet containing 1.0 g / Kg HE continuously for 7 days, followed by a basal diet for 21 days. The HE102 group was fed a diet containing 1.0 g / Kg HE alternately every other day for 14 consecutive days, followed by a basal diet for another 14 days. The HE103 group was intermittently fed a diet containing 1.0 g / Kg HE every 2 days for 21 consecutive days, followed by a basal diet for 7 days. Finally, the HE104 group was regularly fed a diet containing 1.0 g / Kg HE every 3 days for a total of 28 days (see Figure 1 Effect of Aeromonas vermiformis infection on the survival rate of California bass The infection results showed that the survival rate of California bass juveniles increased with the increase of the interval between feeding with HE. Among them, the HSR group fed with 1.0 g / kg HE feed every 3 days showed the highest survival rate (92.0%) (see Figure 2 ).

[0021] In the second phase, 25 juveniles of the same size were housed in the control and HSR groups under the same conditions as in the first phase. All fish were fasted for 24 hours before sampling. Fish were anesthetized with 20 mg / L of diazepam (MS-222). While anesthetized, all fish were collected and weighed. Nine fish were randomly selected from each group, and blood samples were drawn from the caudal vein. Liver tissue was collected for analysis of biochemical parameters and antioxidant capacity. Furthermore, three fish were randomly selected from each replicate group for dissection, and intestinal and liver tissues were histologically examined to assess structure and possible pathological changes. At the beginning and end of the feeding trial, body weight and length of California bass were measured, and weight gain, specific growth rate, and feed conversion ratio were calculated. The results are shown in Table 1. Feeding the compound Chinese herbal medicine significantly affected weight gain, specific growth rate, conditioning factor, liver-to-body ratio, and feed conversion ratio of California bass.

[0022] Table 1 Effects of interval feeding on the growth performance of California bass After the second stage of the feeding experiment, the WGR, SGR, and HSI of the HSR group were significantly higher than those of the control group (p<0.05). Meanwhile, there was no statistically significant difference in FCR and CF between the two groups.

[0023] Notes: Data are presented as mean ± standard error of the mean (SEM); n = 3. Values with different superscripts within the same row are significantly different (p < 0.05). IBW, initial body weight; FBW, final body weight; WGR, body weight gain rate; SGR, specific growth rate; HSI, liver-to-body ratio; FCR, feed conversion ratio; CF, fraction of fatness. Con group, fed a basal diet; HSR group, fed a diet containing 1.0 g / kg HE every 3 days for 28 days.

[0024] Effects of regular feeding of compound Chinese herbal medicine every three days on the antioxidant capacity of California bass. In the liver, the SOD, T-AOC activities and MDA concentrations of the HSR group were significantly decreased compared with the control group (p < 0.05) (see Figure 3 A). At the same time, the change trends of serum AST and ALT activities were similar to those of the above indicators, indicating that appropriate feeding intervals can alleviate the negative impact of 1.0 g·Kg-1 HE in the feed on liver health (see Figure 3 B).

[0025] Liver and intestinal morphology In the control group and HSR group, hepatocytes were neatly arranged and the hepatic cords were evenly arranged. At the same time, a certain degree of vacuolation and nuclear displacement were observed in both groups. Morphological analysis showed that the intestinal mucosal folds, muscular layer and lamina propria were intact in both groups. However, an increase in villus height was detected in the HSR group (p < 0.05) ( Figure 4 , Table 2).

[0026] Table 2. Effects of herbal extracts on intestinal tissue parameters of largemouth bass Note: Data are presented as mean ± standard error of the mean (SEM; n = 3). Different superscript values in the same row are significantly different (p < 0.05). Con group, fed with basal diet; HSR group, fed with 1.0 g·kg-1 herbal combination. The fish with the highest survival rate after challenge with Aeromonas vernix in phase I In summary, the HSR group (fed 1.0 g / Kg HE every 3 days) had the highest survival rate and significantly improved the specific growth rate. At the same time, the results showed that this feeding method did not damage the liver. Studies have shown that this feeding method can slow down the damage caused by high-dose HE to California bass.

[0027] The above embodiments describe in detail the structure, features and effects of the present invention. The above are only preferred embodiments of the present invention. Any changes made in accordance with the concept of the present invention, or modifications to equivalent embodiments with equivalent changes, which do not exceed the scope covered by the specification, should be within the scope of protection of the present invention.

Claims

1. A method for preparing a composite Chinese herbal medicine additive for California sea bass, characterized in that: The following steps are involved: (1) Soak the Chinese herbal medicines in 10 times the weight of water for 3 hours, then decoct for 40 minutes and filter out the medicinal liquid; add 30 times the weight of water to the filtered medicinal residues, decoct for another 40 minutes and filter out the medicinal liquid; the Chinese herbal medicines are scutellaria baicalensis, dandelion, sappan wood and pulsatilla chinensis; (3) Combine the two decoctions; (4) Concentrating the above medicinal solution to a crude drug content of 1 g / mL; (5) 20 parts by weight of Scutellaria baicalensis concentrated solution, 27 parts by weight of Taraxacum mongolicum concentrated solution, 25 parts by weight of Sappan wood concentrated solution, and 15 parts by weight of Pulsatilla chinensis concentrated solution were mixed to obtain a California sea bass compound Chinese herbal medicine additive.

2. A compound Chinese herbal medicine additive for California sea bass prepared by the preparation method according to claim 1.

3. A method for periodically applying the compound Chinese herbal medicine additive for California sea bass according to claim 2, characterized in that: The California bass was fed a compound feed every three days for 28 days; the compound feed contained 0.1 wt% of a compound Chinese herbal medicine additive; the compound Chinese herbal medicine additive contained 20 parts by weight of Scutellaria baicalensis concentrate, 27 parts by weight of Taraxacum mongolicum concentrate, 25 parts by weight of Sappanwood concentrate and 15 parts by weight of Pulsatilla scabra concentrate; the crude drug content in the concentrate was 1 g / mL.

4. The method according to claim 3, characterized in that The preparation method of the California bass compound feed is as follows: spraying the California bass compound Chinese herbal medicine additive at a rate of 0.1 wt % onto the California bass basic feed.

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