Compound Chinese herbal medicine and aquatic animal prefabricated feed

Abstract

The invention provides a compound Chinese herbal medicine and aquatic animal prefabricated feed. The compound Chinese herbal medicine is prepared from the following components in parts by weight: 20-30 parts of sweet wormwood herb, 10-15 parts of radix astragali, 10-15 parts of radix isatidis, 5-10 parts of radix bupleuri, 5-10 parts of rhizoma polygonati, 5-10 parts of radix notoginseng, 5-10 parts of fructus forsythiae, 5-8 parts of sedum sarmentosum, 5-10 parts of radix salviae miltiorrhizae and 10-15 parts of liquorice root. The aquatic animal prefabricated feed contains composite Chinese herbal medicines; when the feed is prepared, the composite Chinese herbal medicines are directly added into feed raw materials to be processed and prepared into the feed. The metabolic level of livers (or hepatopancreas) of aquatic animals is regulated through the compound Chinese herbal medicines, the problem of various diseases caused by constitution decline due to low immunity caused by obesity of the aquatic animals due to excessive addition of energy substances in the artificial compound feed is solved, the immunity of the animals is improved, and the anti-stress and disease-resistant capabilities are enhanced.

Description

Technical Field

[0001] The invention relates to aquatic animal breeding, and in particular to a composite Chinese herbal medicine and application thereof, aquatic animal prefabricated feed and a preparation method thereof. Background Art

[0002] As global awareness of food safety and environmental protection continues to increase, the aquaculture industry faces new challenges and opportunities, and is also actively exploring a more sustainable and healthy development path. The high-density intensive breeding of aquatic products, the decline in resistance caused by the degeneration of germplasm, and the deterioration of water quality caused by the large-scale use of artificial compound feeds often lead to large-scale outbreaks of fishery diseases. The fishery is helpless in the face of frequent viral and bacterial diseases, causing huge losses. For example, the viral hemorrhagic disease of silver crucian carp has made crucian carp almost impossible to raise, and the iridovirus of sea bass has caused regional annihilation. The hepatopancreatic necrosis syndrome of Chinese mitten crab has caused a significant reduction in production, and the aquaculture industry has suffered an unprecedented heavy blow. The impact of diseases has caused breeders to abuse drugs, and the problem of excessive drug residues and drug resistance of antibiotics has become increasingly serious. This not only makes it more difficult to treat fishery diseases, but also increases the metabolic burden of aquatic animals, damages their health, and increases breeding costs and risks, posing a potential threat to product quality and food safety. Summary of the invention

[0003] Research has found that aquatic animals are mainly raised with artificial compound feeds. Aquatic animals have a higher demand for protein than livestock and poultry, and many species use 30-40% of the protein, which makes the feed cost high. In production, low-end, indigestible protein sources such as cottonseed meal and feather meal are often used as raw materials. In addition, in order to save protein, high-energy feeds such as fat are added to seek better growth. The oils used in feed production are often leftovers from other industries and have high oxidation levels. After ingestion, aquatic animals often become obese, have severe fatty liver, and even suffer from liver damage (such as hepatopancreatic necrosis syndrome in Chinese mitten crabs). After metabolic abnormalities, the physical condition of aquatic animals drops sharply, their immune resistance is low, and various diseases often break out. Many diseases of aquatic animals are conditional pathogens. When the immunity of animals decreases and the environment deteriorates, they appear one after another. It is almost impossible for drugs to eradicate the threat of pathogens, making disease control more difficult.

[0004] Based on this, the present invention proposes a composite Chinese herbal medicine and aquatic animal prefabricated feed, aiming to seek a solution from the theory of traditional Chinese medicine (starting from the concept of preventing diseases before they occur in traditional Chinese medicine).

[0005] The technical solution of the present invention: A composite Chinese herbal medicine is prepared from the following components in parts by weight: 20-30 parts of Artemisia annua, 10-15 parts of Astragalus, 10-15 parts of Isatis indigotica, 5-10 parts of Bupleurum, 5-10 parts of Polygonatum, 5-10 parts of Panax notoginseng, 5-10 parts of Forsythia, 5-8 parts of Creeping Herb, 5-10 parts of Salvia miltiorrhiza, and 10-15 parts of Licorice.

[0006] A prefabricated feed for aquatic animals, comprising the compound Chinese herbal medicine according to claim 1; when preparing the feed, the compound Chinese herbal medicine is directly added to the feed raw materials to be processed together to prepare the feed.

[0007] The above technical solution is further refined. Before making the feed, the compound Chinese herbal medicine is dried and ultra-finely ground into fine powder with a particle size of less than 10 μm, and then the fine powder is put into a blender and mixed for 30 minutes to be fully mixed.

[0008] A prefabricated feed for aquatic animals, wherein the raw material ratio of the prefabricated feed is: 5% Chinese medicine formula package, 11% fish meal, 12% cottonseed meal, 5% meat and bone meal, 14% soybean meal, 12% vegetable cake, 16% flour, 16.5% bran, 4% vegetable oil, 4% minerals, and 0.5% squid paste; the Chinese medicine formula package is made of the composite Chinese herbal medicine according to claim 1.

[0009] The above technical solution is further refined. The preparation of the Chinese medicine formula package is specifically as follows: the composite Chinese herbal medicine is dried and ultrafinely ground into fine powder with a particle size of less than 10 μm, and then the fine powder is put into a blender and mixed for 30 minutes to be fully mixed.

[0010] The above technical solution is further refined. The preparation of the prefabricated feed is specifically: the raw materials are mixed evenly, heated to above 85° C. with high-temperature steam for 30 minutes, granulated, cooled, and bagged.

[0011] The above-mentioned prefabricated feed for aquatic animals is easy to produce, saves the need for pre-feeding treatment of commercial feed, and is convenient to use; The above-mentioned prepared feed for aquatic animals can be used for a long time or intermittently during the period of high incidence of diseases. It can prevent the outbreak of fishery diseases in aquatic animals in spring and autumn, improve the anti-stress level, and significantly improve the mortality rate after the outbreak of diseases. The above-mentioned prepared feed for aquatic animals is pure Chinese medicinal plant feed, which is safe, has no toxic side effects, and does not produce drug resistance.

[0012] In the preparation process of the above-mentioned aquatic animals, the Chinese medicine is fully crushed by ultrafine grinding technology, and then the maturation process and the market are added in the feed production process to make the medicine evenly dispersed in the feed, and the processing process is completed.

[0013] Preferably, an ultrafine pulverizer is used to pulverize the dried composite Chinese herbal medicine into fine powder with a particle size of less than 10 μm; the ultrafine pulverizer is improved, the ultrafine pulverizer comprises a body, a feed hopper is provided on the top of the body, a crushing roller group is installed at the bottom of the feed hopper, a refining mechanism is provided at the bottom of the crushing roller group, and a containing mechanism is provided inside the body; the refining mechanism comprises a first guide plate, a second guide plate and a third guide plate, two refining knife rollers are provided between the first guide plate and the second guide plate and between the second guide plate and the third guide plate, the ends of the two refining knife rollers are fixedly connected to a support shaft, the two support shafts are fixedly connected to a transmission gear, one end of the support shaft is fixedly connected to a drive motor, a fixed seat is provided on one side of the two refining knife rollers, a rotating shaft is rotatably connected inside the fixed seat, the rotating shaft and an end of a support shaft are fixedly connected to a synchronous wheel, and the two A synchronous belt is sleeved between the synchronous wheels, and a plurality of cams are fixedly connected to the rotating shaft, and a vertical frame is provided on the top of the cam, and a push rod is slidably connected inside the vertical frame, and a limit block is fixedly connected to the push rod, and a return spring is provided between the limit block and the vertical frame; the first guide plate, the second guide plate and the third guide plate are all fixedly installed on the machine body, and the first guide plate, the second guide plate and the third guide plate are all inclinedly arranged on the inner wall of the machine body; the two thinning knife rollers are connected by transmission gears, and the thinning knife rollers are rotatably connected to the inner wall of the machine body through a support shaft; the fixed seat is fixedly connected to the inner wall of the machine body, and the two fixed seats are respectively arranged at the bottom of the first guide plate and the second guide plate; the vertical frame is fixedly installed on the top of the fixed seat, and the bottom end of the push rod contacts the cam; one end of the return spring is fixedly connected to the bottom surface of the vertical frame, and the other end of the return spring is fixedly connected to the top surface of the limit block.

[0014] When in use, the (dried) composite Chinese herbal medicine is first crushed by the crushing roller group, and then falls along the first guide plate, the second guide plate and the third guide plate in sequence. During the falling process, the driving motor simultaneously drives the refinement knife roller to rotate, refines and crushes the composite Chinese herbal medicine, and knocks the first guide plate and the second guide plate to produce vibration, so that the composite Chinese herbal medicine can fall fully to avoid residue, so that it can be crushed and refined multiple times, effectively improving the crushing effect.

[0015] The above technical solution is further refined, the containing mechanism includes a containing box, a containing cavity is provided inside the containing box, and the containing box is plugged into the machine body; a handle is fixedly connected to the front surface of the containing box, and an adsorption magnet is fixedly connected to the rear surface of the containing box. By inserting the containing box into the machine body, the fully crushed and refined composite Chinese herbal medicine is received and fixed by magnetic attraction with the adsorption magnet, thereby improving the stability of use.

[0016] The invention has the advantages of reasonable design and ingenious conception. It regulates the metabolic level of aquatic animal liver (or hepatopancreas) through compound Chinese herbal medicine, improves the problem of low immunity caused by excessive addition of energy substances in artificial compound feed, resulting in a decline in physical fitness and outbreak of various diseases, improves animal immunity, and enhances anti-stress and disease resistance. By adopting a special processing technology, Chinese medicine processing is realized in the process of feed processing to make functional feed that improves metabolism and improves animal immunity, helping fisheries to achieve healthy breeding without antibiotics; Artemisia annua, bupleurum, astragalus, Panax notoginseng, forsythia, scutellaria baicalensis, isatis indigotica, polygonatum, salvia miltiorrhiza and licorice are applied to the prevention and treatment of aquatic animal diseases, especially after adding Artemisia annua, the control effect on invisible parasites of aquatic animals is more significant, bupleurum, forsythia and licorice are more significant in the control of bacteria, isatis indigotica, astragalus and licorice are more significant in the control of viruses, and scutellaria baicalensis, Panax notoginseng, polygonatum, astragalus and licorice have significant effects on the metabolic regulation of liver (hepatopancreas) and spleen. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a comparative picture of the hepatopancreas histology of Chinese mitten crabs fed different diets.

[0018] Figure 2 Comparative pictures of midgut histology of shrimp fed different diets (low magnification 200× for intestinal histology and high magnification 400× for group intestinal histology).

[0019] Figure 3 This is a graph showing the expression levels of immune-related genes in the gill tissue of Penaeus vannamei fed different diets.

[0020] Figure 4 This is a comparison chart of the computational classification units of the intestinal microbial flora of Penaeus vannamei fed with different feeds.

[0021] Figure 5 This is a diagram of the bacterial abundance at the phylum level in the intestinal microorganisms of Penaeus vannamei under different feeding conditions.

[0022] Figure 6 This is a diagram of the bacterial abundance at the genus level in the intestinal microorganisms of Penaeus vannamei under different feeding conditions.

[0023] Figure 7 This is a comparison of the differences in the intestinal microbial flora at the phylum level of Penaeus vannamei fed with different feed conditions.

[0024] Figure 8 This is a comparison of the differences in the intestinal microbial flora at the phylum level of Penaeus vannamei fed with different feed conditions.

[0025] Fig. 9 It is a schematic diagram of the overall structure of the ultrafine pulverizer.

[0026] Fig.10 It is a schematic diagram of the refinement mechanism structure of the ultrafine pulverizer.

[0027] Fig.11 It is a schematic diagram of the fixed seat structure of the ultrafine pulverizer.

[0028] Fig.12 It is a schematic diagram of the structure of the loading mechanism of the ultrafine pulverizer.

[0029] In the figure: 1. machine body; 2. refining mechanism; 201. first guide plate; 202. second guide plate; 203. third guide plate; 204. refining knife roller; 205. supporting shaft; 206. transmission gear; 207. fixing seat; 208. rotating shaft; 209. synchronous wheel; 210. synchronous belt; 211. cam; 212. stand; 213. push rod; 214. return spring; 215. limit block; 3. feed hopper; 4. filling mechanism; 401. filling box; 402. handle; 403. filling cavity; 404. adsorption magnet; 5. crushing roller group. DETAILED DESCRIPTION

[0030] Example 1: Chinese medicine pre-made feed for disease prevention and control of silver carp and bighead carp in spring green grass ponds Traditional Chinese medicine formula package: dry 30 parts of Artemisia annua, 10 parts of Astragalus, 10 parts of Radix Isatidis, 10 parts of Bupleurum, 10 parts of Forsythia, 5 parts of Polygonatum, 5 parts of Panax notoginseng, 5 parts of Herba Creepersii, 5 parts of Salvia miltiorrhizae, and 10 parts of Licorice, and then ultrafine grind them into fine powder with a particle size of less than 10 μm, and then put the fine powder into a blender and mix and stir for 30 minutes to mix thoroughly; Prefabricated feed: Evenly mix 5% of Chinese medicine formula package, 11% of fish meal, 12% of cottonseed meal, 5% of meat and bone meal, 14% of soybean meal, 12% of vegetable cake, 16% of flour, 16.5% of bran, 4% of vegetable oil, 4% of minerals and 0.5% of squid paste, heat to above 85℃ with high-temperature steam for 30 minutes, granulate, cool and bag.

[0031] Example 2: Pre-made feed with Chinese medicine for disease prevention and control in spring ponds with grass carp, silver carp and bighead carp as the main culture Traditional Chinese medicine formula package: dry 20 parts of Artemisia annua, 15 parts of Astragalus, 15 parts of Radix Isatidis, 10 parts of Bupleurum, 10 parts of Forsythia, 5 parts of Polygonatum, 5 parts of Panax notoginseng, 5 parts of Herba Creepersii, 5 parts of Salvia miltiorrhizae, and 10 parts of Licorice, and then ultrafine grind them into fine powder with a particle size of less than 10 μm, and then put the fine powder into a blender and mix and stir for 30 minutes to mix thoroughly; Prefabricated feed: Mix 5% of Chinese medicine formula package, 18% of fish meal, 10% of cottonseed meal, 6% of meat and bone meal, 20% of soybean meal, 16% of vegetable cake, 16% of flour, 4.5% of vegetable oil, 4% of minerals, and 0.5% of squid paste, heat to above 85℃ with high-temperature steam for 30 minutes, granulate, cool, and bag; Example 3: Pre-made feed with Chinese medicine for disease prevention and control in a mixed culture pond of grass carp, crucian carp, silver carp and bighead carp with bream as the main species

[0032] Traditional Chinese medicine formula package: dry 25 parts of Artemisia annua, 15 parts of Astragalus, 10 parts of Radix Isatidis, 10 parts of Radix Bupleuri, 5 parts of Fructus Forsythiae, 5 parts of Polygonatum, 5 parts of Notoginseng, 8 parts of Herba Creepersii, 5 parts of Salvia miltiorrhizae, and 12 parts of Radix Glycyrrhizae, and grind them into fine powder with a particle size of less than 10 μm, then put the fine powder into a blender and mix and stir for 30 minutes to mix thoroughly; Prefabricated feed: Mix 4% of Chinese medicine formula package, 10% of fish meal, 16% of cottonseed meal, 6% of meat and bone meal, 17% of corn meal, 21% of vegetable cake, 17% of flour, 4.5% of vegetable oil, 4% of minerals, and 0.5% of squid paste, heat to above 85℃ with high-temperature steam for 30 minutes, granulate, cool, and bag; Example 4: Pre-made feed with Chinese medicine for disease prevention and control in ponds with sea bass as the main species, crucian carp, silver carp and bighead carp

[0033] Chinese medicine formula package: 20 parts of Artemisia annua, 14 parts of Astragalus, 14 parts of Isatis, 9 parts of Bupleurum, 9 parts of Forsythia, 7 parts of Polygonatum, 6 parts of Panax notoginseng, 6 parts of Creeping Herb, 5 parts of Salvia miltiorrhiza, 10 parts of Licorice are dried and ultrafinely ground into fine powder with a particle size of less than 10 μm, and then the fine powder is put into a blender and mixed for 30 minutes to mix thoroughly.

[0034] Prefabricated feed: evenly mix 3% of Chinese medicine formula package, 32% of fish meal, 16% of meat and bone meal, 15% of flour, 19% of soybean meal, 5% of squid paste, 8% of fish oil and soybean oil, 1.5% of monocalcium phosphate, 0.1% of multivitamins and 0.4% of multiminerals, heat to above 85℃ with high-temperature steam for 30 minutes, granulate, cool and bag. Example 5: Chinese medicine pre-made feed for disease prevention and control in river crab ponds

[0035] Traditional Chinese medicine formula package: dry 20 parts of Artemisia annua, 15 parts of Astragalus, 10 parts of Forsythia, 10 parts of Polygonatum, 10 parts of Isatis, 10 parts of Bupleurum, 5 parts of Panax notoginseng, 5 parts of Herba Salviae Miltiorrhizae, 5 parts of Licorice into fine powder with a particle size of less than 10 μm, then put the fine powder into a blender and mix for 30 minutes to mix thoroughly; Prefabricated feed: 2% Chinese medicine formula, 12% fish meal, 20% snail meat, 15% flour, 20% soybean meal, 16% corn flour, 5% squid paste, 8% fish oil and soybean oil, 1.5% calcium dihydrogen phosphate, 0.1% multivitamin, and 0.4% multimineral. Mix evenly, heat to above 85℃ with high-temperature steam, mature for 30 minutes, granulate, cool, and bag. Implementation Case 6. Pre-made feed with traditional Chinese medicine for disease prevention and control in white shrimp ponds

[0036] Chinese medicine formula package: 20 parts of Artemisia annua, 13 parts of Astragalus, 10 parts of Forsythia, 10 parts of Polygonatum, 10 parts of Isatis, 10 parts of Bupleurum, 5 parts of Panax notoginseng, 7 parts of Creeping Herb, 5 parts of Salvia miltiorrhiza, and 10 parts of Licorice are dried and ultrafinely ground into fine powder with a particle size of less than 10 μm, and then the fine powder is put into a blender and mixed for 30 minutes to fully mix to make a Chinese medicine formula package.

[0037] Prefabricated feed: Mix 2% of Chinese medicine formula package, 34% of Peruvian fish meal, 18% of white fish meal, 4% of yeast powder, 6% of soybean lecithin, 14% of soybean meal, 7.3% of krill meal, 2.5% of seaweed powder, 6.5% of second flour, 5.2% of wheat gluten powder and 0.5% of salt evenly, heat to above 85℃ with high-temperature steam for 30 minutes, granulate, cool and bag. Experiment 1: Study on the immunity test of white shrimp

[0038] The experiment was divided into three treatment groups, each with four parallels. The three groups of feed were fed to Penaeus vannamei for 8 weeks. Through mid-term and late sampling, various indicators were tested to determine the effects of Chinese herbal medicine added feed on the growth performance, nonspecific immunity and intestinal health of Penaeus vannamei. The three groups were the Commercial feed group (CF), the Chinese herbs added to the commercial feed group (ChCF) and the Interval feeding group (IF) with two feeds one week apart.

[0039] The commercial Chinese herbal medicine feed is prepared by adding 2% of a Chinese herbal formula package to ordinary commercial feed (20 parts of Artemisia annua, 13 parts of Astragalus, 10 parts of Forsythia, 10 parts of Polygonatum, 10 parts of Isatis, 10 parts of Bupleurum, 5 parts of Panax notoginseng, 7 parts of Creeping Herb, 5 parts of Salvia miltiorrhiza and 10 parts of Licorice are dried and ultrafinely ground into fine powder with a particle size of less than 10 μm, and then the fine powder is put into a blender and mixed for 30 minutes to mix thoroughly), 2% of the Chinese herbal formula package, 34% of Peruvian fish meal, 18% of white fish meal, 4% of yeast powder, 6% of soybean lecithin, 14% of soybean meal, 7.3% of krill meal, 2.5% of seaweed powder, 6.5% of second flour, 5.2% of wheat gluten powder and 0.5% of salt are mixed evenly, heated to above 85°C with high-temperature steam for 30 minutes, granulated, cooled and bagged.

[0040] The feed formula of the control group was: 2% second-rate flour, 34% Peruvian fish meal, 18% white fish meal, 4% yeast powder, 6% soybean lecithin, 14% soybean meal, 7.3% krill meal, 2.5% seaweed powder, 6.5% second-rate flour, 5.2% wheat gluten powder, and 0.5% salt. 1.1 Experimental shrimp and feeding management

[0041] The shrimp fry of Penaeus vannamei were temporarily raised in the earth pond and fed with commercial feed. Feeding was stopped 24 hours before the formal experiment, and 600 healthy shrimps of uniform size (initial weight was about 3.0g) were selected and randomly distributed into 12 circulating water culture barrels (the volume, water depth and diameter of the culture barrels were 450L, 1m and 0.8m respectively). The experiment set up 3 treatments in total, with 4 replicates in each treatment. During the culture period, the daily feeding amount was 3% to 10% of the shrimp body weight (high feeding in the early stage and low feeding in the later stage), and it was fed in 3 times (8:00, 16:00 and 22:00). The specific feeding amount was adjusted in time according to the feeding situation of the shrimp and the weather conditions, so that the feeding amount of each barrel was basically the same, and it was ensured that the feed was eaten within 2 hours after feeding; it was aerated day and night, and the feces at the bottom of the culture barrel was sucked away by siphoning. The sewage was sucked once a week in the early stage of culture, and 1 / 3 of the water was changed. The sewage was sucked and the water was changed twice a week in the later stage of culture. The water source used was the pond water after sedimentation. During the test, the water temperature was 28-33°C, the salinity was 0.5‰-1.0‰, and the dissolved oxygen was ≥5.6mg / l -1 , pH value 7.8~8.5, ammonia nitrogen ≤0.2mgl -1 , nitrite ≤ 0.05mgl -1 The feeding experiment lasted for 56 days. The breeding experiment was carried out at the breeding base. 1.2 Sample collection

[0042] Before the start of the breeding experiment, 30 shrimps were randomly selected and stored in a -20℃ refrigerator, of which 10 shrimps were combined into one sample for initial sample composition analysis. After the end of the breeding experiment, feeding was stopped for 24 hours, and the shrimps in each breeding bucket were weighed and the number of shrimps was recorded; 4 shrimps were randomly selected from each breeding bucket, the surface moisture of the shrimps was sucked dry, and stored in a -20℃ refrigerator for whole shrimp body composition analysis. 9 shrimps were randomly selected from each breeding bucket, and hemolymph was extracted from the pericardial cavity behind the cephalothorax. After mixing with anticoagulant in a ratio of 1:1, centrifuged at 10000rmin-1 for 20min, and the supernatant was stored at -80℃ for the detection of immune and antioxidant indicators (the anticoagulant formula was 0.14M sodium chloride, 0.1M glucose, 26mM citric acid, 30mM sodium citrate, and 10mM EDTA). After blood was collected from the shrimp, the hepatopancreas (3 shrimps were combined into one sample) was taken and frozen at -80℃ for the determination of relevant digestive enzyme activities. Three emptied-intestine shrimps were collected from each culture tank, and the midgut and complete hepatopancreas of the shrimp were preserved in Bouin's fixative. Six emptied-intestine shrimps were collected from each culture tank, and the body surface was wiped with 75% alcohol. The midguts were combined into one sample, placed in liquid nitrogen for 6 hours, and then stored at -80°C for intestinal microbial analysis. 1.3 Growth performance

[0043] Growth performance indicators include survival rate (SR), weight gain rate (WG), feed conversion rate (FCR), protein deposition rate (PRE) and fat deposition rate (LRE): Survival rate, (%) = 100 × (number of shrimps harvested / number of shrimps released) Weight gain rate, (%) = 100 × (final average mass - initial average mass) / initial average mass Feed coefficient = feeding amount / (final average mass-initial average mass) Protein deposition rate, (%) = 100 × (final average mass × final whole shrimp crude protein content - initial average mass × initial whole shrimp crude protein content) / (feeding amount × feed crude protein content) Fat deposition rate, (%) = 100 × (final average mass × final whole shrimp crude fat content - initial average mass × initial whole shrimp crude fat content) / (feeding amount × feed crude fat content) 1.4 Serum biochemical indexes and hepatopancreatic digestive enzyme activities Acid phosphatase (ACP), alkaline phosphatase (AKP) and total antioxidant capacity (T-AOC) were determined by colorimetry. Lysozyme (LZM) was determined by turbidimetry. Catalase (CAT) was determined by complex method. Total superoxide dismutase (T-SOD) was determined by xanthine oxidase method. Malondialdehyde (MDA) was determined by thiobarbituric acid method. Glutathione peroxidase (GSH-PX) was determined by nitrobenzoic acid method. The above indicators were determined according to the instructions of the kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, Jiangsu). 1.5 Intestinal and hepatopancreatic tissue sections

[0044] When making slices, the midgut and hepatopancreas were dehydrated with ethanol, transparentized with xylene, embedded in paraffin, sliced ​​(thickness 5 μm) (LeiknRM2235 slicer, Germany), stained with HE, and sealed to make permanent sections. The morphological characteristics of intestinal folds were observed and photographed under a light microscope (NikonYS100 microphotography system), and the height, width and muscle thickness of intestinal epithelial folds were measured with a micrometer. 1.6 Immune gene expression in gill tissue

[0045] The specific primer pairs for the β-actin gene and other genes (Table 1) were based on the relevant cDNA sequences of Penaeus vannamei in the National Center for Biotechnology Information (NCBI) database. Total RNA was extracted using Trizol reagent, and the concentration and purity of total RNA were determined by UV spectrophotometer and agarose gel electrophoresis. The TaKaRa reverse transcription kit (PrimeScript TMTotal RNA was reverse transcribed into cDNA by RT. The fluorescence quantitative PCR reaction procedure was as follows: 95℃ pre-denaturation for 3min, 95℃ denaturation for 10s, 60℃ annealing for 10s, 72℃ extension for 30s, a total of 39 PCR cycles, and finally a melting curve test. The gene expression level of gill tissue of Penaeus vannamei in the NM group was used as a benchmark, and β-actin was used as an internal reference gene. The 2-ΔΔCt method was used to calculate the expression level of immune-related genes in gill tissues of each group. Table 1 Real-time quantitative PCR primers for immune-related genes and β-actin in Litopenaeus vannamei.

[0046] 1.7 Gut microbiota based on 16s rRNA sequencing

[0047] Four shrimps were taken from each parallel group, and the body surface was wiped with 75% alcohol for sterilization and disinfection. The intestines were taken under sterile conditions and placed in 1.0 ml centrifuge tubes for intestinal microbial composition analysis. The sequencing experiment was commissioned to Shanghai Meiji Biomedical Technology Co., Ltd. (China) for next-generation sequencing (NGS) library preparation and Illumina Miseq sequencing. Primers 338F (5'-ACTCCTACGGGAGGCAGCAG-3') and 806R14.47 (5'-GGACTACHVGGGTWTCTAAT-3') were used to amplify the V3-V4 hypervariable region of bacterial 16S rRNA (GeneAmp9700, ABI, USA), and the microbial community was identified by 16S rRNA sequence analysis. 1.8 Data Processing

[0048] Data were expressed as mean ± standard deviation (SD). SPSS 22.0 statistical software was used for statistical analysis. All data were analyzed using one-way analysis of variance (ANOVA). If there was a significant difference, the Duncan multivariate range test was used to determine the statistical significance between the groups. P When the value was <0.05, the mean values ​​were considered to be significantly different. 2. Results

[0049] 2.1 Growth performance P ≤0.05). There was no significant difference in survival rate, protein deposition rate, and fat deposition rate among the groups. At the end of the breeding period, the average weight, weight gain rate, and survival rate of the ChCF-L group were significantly higher than those of the CF-L group and the IF-L group, and the feed coefficient was significantly lower than that of the CF-L group and the IF-L group. The survival rate of the ChCF-L group and the IF-L group was significantly higher than that of the CF-L group ( P ≤0.05). There was no significant difference in the protein deposition rate and fat deposition rate of shrimps among the groups ( P ≥0.05). Table 2. Effects of different feeds on the growth performance of Penaeus vannamei

[0050] 2.2 Serum biochemical indexes

[0051] As shown in Table 3, in the middle of the culture period, the activities of serum ACP, ALP, T-AOC, GSH-Px and T-SOD in the ChCF-I group were significantly higher than those in the CF-I group, and the serum MDA content was significantly lower than that in the CF-I group ( P ≤0.05). The serum LZM activity of shrimp in ChCF-I and IF-I groups was significantly higher than that in CF-I ( P ≤0.05). At the end of the culture period, the serum ACP, ALP, LZM, and T-AOC activities of ChCF-L and IF-L groups were significantly higher than those of the CF-L group, and the serum MDA content was significantly lower than that of the CF-L group ( P ≤0.05). The serum GSH-Px and T-SOD activities of shrimps in the ChCF-L group were significantly higher than those in the CF-L group ( P ≤0.05). There was no significant difference in the serum CAT activity of shrimps among the groups ( P ≥0.05). Table 3. Effects of different feeds on serum biochemical parameters of Penaeus vannamei

[0052] 2.3 Digestive enzyme activity

[0053] In the middle of the culture period, the activities of hepatopancreatic protease and lipase in the ChCF-M group were significantly higher than those in the CF-M group ( P ≤0.05). There was no significant difference in the hepatopancreatic amylase activity of shrimps among the groups ( P ≥0.05). At the end of the culture period, there was no significant difference in the activities of hepatopancreatic protease, lipase and amylase in each group of shrimp ( P ≥0.05) (Table 4).

[0054] Table 4. Effects of different feeds on hepatopancreatic digestive enzymes in Litopenaeus vannamei

[0055] 2.4 Hepatopancreas and Intestinal Tissue Morphology

[0056] Figure 1It can be seen that in the mid-term and late sampling, the epithelial cells between the hepatopancreatic tubules in the Chinese herbal feed supplement group and the alternating feeding group were closely connected, the hepatopancreatic tubules showed a star-shaped polygonal structure, clear morphology, and moderate lumen size and epithelial thickness. In the mid-term and late sampling of the common commercial feed group, the star-shaped polygonal structure of some hepatopancreatic tubules disappeared, the lumen became larger, the epithelium became thinner, and the B cells and r cells were compressed into an ellipse. The hepatopancreas of normal vannamei shrimp is composed of hepatic tubules, and the center of the hepatic tubule is the lumen. Each hepatic tubule is composed of a single layer of epithelial cells located on the basement membrane. The epithelial cells are divided into embryonic cells (E cells), which are undifferentiated cells distributed at the blind end (top) of the hepatic tubules, secretory cells (B cells), fiber cells (F cells) and storage cells (R cells). R (restzellen) cells usually contain lipid vacuoles of varying sizes (R cells show typical characteristics of absorptive epithelium. These R cells are slender and seem to be "squeezed" between other types of cells, but extend to the entire epithelial cell layer). F (fibrous) cells contain numerous ribosomes and a well-developed endoplasmic reticulum; B (blasenzellen) cells typically contain a single large secretory vesicle.

[0057] Figure 2 The midgut morphology of shrimp fed different feeds at different stages is shown. In the midgut of shrimps in the Chinese herbal medicine feed group and the alternating tail casting group, villi formed by a single layer of columnar cells were clearly visible, and the villi were arranged in an orderly and compact manner. In the mid-stage of the shrimps in the ordinary commercial feed group, the height of the villi and the thickness of the intestinal wall decreased significantly, and in the late stage, there was even only a single layer of columnar cells on the inner wall of some midguts, and no obvious midgut folds were observed.

[0058] As shown in Table 5, at the mid-term sampling, the number of intestinal villi in the ChCF-I and IF-I groups was significantly greater than that in the CF-I group ( P ≤0.05), there were no significant differences in villus height, villus width and base thickness among the groups ( P ≥0.05). At the end of the sampling period, the number and height of intestinal villi in the ChCF-I and IF-I groups were significantly greater than those in the CF-I group ( P ≤0.05), there was no significant difference in villus width and base thickness among the groups ( P ≥0.05). Table 5. Effects of different feeds on the intestinal tissue morphology of Penaeus vannamei

[0059] 2.5 Immune gene expression in gill tissue

[0060] Depend on Figure 3It can be seen that in the middle of the culture period, the expression levels of Lysozyme and Toll-like receptor genes in the gill tissues of the ChCF-I and IF-I groups were significantly higher than those in the CF-I group, and the expression levels of IMD and SOD genes in the gill tissues of the ChCF-I group were significantly higher than those in the CF-I group ( P ≤0.05). At the end of the culture period, the expression levels of Lysozyme and IMD genes in the gill tissues of the ChCF-L and IF-L groups were significantly higher than those in the CF-L group, and the expression levels of Toll-like receptor and SOD genes in the gill tissues of the ChCF-L group were significantly higher than those in the CF-L group ( P ≤0.05). 2.6 Gut Microbes

[0061] Through 16SrRNA sequencing, a total of 1933757 valid sequences were obtained, with bases of 813334348bp and an average sequence length of 420bp, of which 85.63% of the sequences were between 421 and 440bp. After cluster analysis of the obtained sequences, 520, 1170, 1161, 656, 869 and 1120 representative bacterial OTUs were obtained in the six groups, respectively. There were 184 common OTUs in the six treatment groups, and 96, 618, 291, 100, 128 and 171 unique OTUs, respectively. Figure 4 shown.

[0062] The dominant flora in the intestinal flora of Penaeus vannamei in each treatment group were Proteobacteria, Firmicutes, Actinobacteriota, Bacteroidota, Patescibacteria, and Cyanobacteria, accounting for 96.59%, 84.50%, 95.95%, 96.73%, 94.46%, and 96.87% of the intestinal flora of Penaeus vannamei, respectively. The rest were mainly unclassified_k__norank_d__Bacteria, Chloroflexi, Acidobacteriota, Verrucomicrobiota, Planctomycetota, etc. Figure 5 shown.

[0063] At the genus level, the dominant bacterial flora in the intestinal tract of Penaeus vannamei in each treatment group were Candidatus_Bacilloplasma, Chitinibacter, Shewanella, Sphingomonas, Vibrio, Aeromonas, and Pseudomonas. Figure 6 shown.

[0064] There were significant differences in the composition and abundance of intestinal microbial phyla among the treatment groups. There were significant differences in the bacterial phyla ( P <0.05), such as Figure 7 shown.

[0065] There were significant differences in the composition and abundance of intestinal microorganisms at the genus level among the treatment groups. There were significant differences in the genera such as Shewanella, Chloroflexi, Pseudorhodobacter, Flavobacterium, Timonella, and Gemmobacter ( P <0.05), such as Figure 8 shown.

[0066] Experiment 2: Effect of Chinese medicine pre-prepared feed on the growth and immune-related enzyme activities of pale white hepatopancreas crab seedlings

[0067] 1 Materials and Methods 1.1 Selection of experimental crab seedlings Catch crab fry in the pond under the cover, use bent tweezers to slightly separate the crab navel to avoid damaging the crab, observe the color of the hepatopancreas at the transparent junction of the crab navel and the rear end of the crab body, and select crab fry with pale pink and white hepatopancreas as experimental crabs. The pale pink and white hepatopancreas at the transparent junction of the crab navel and the rear end of the crab body is consistent with the pale pink and white hepatopancreas in the crab body cavity ( Figure 1 ). A total of 200 crab fry were selected, with an average weight of 7.06 grams.

[0068] Note: The color of the hepatopancreas at the transparent part where the navel on the back of the crab connects to the rear end of the crab body is consistent with the color of the hepatopancreas in the body cavity 1.2 Feed Prefabricated feed with Chinese medicine: evenly mix 2% of Chinese medicine formula package (20 parts of Artemisia annua, 15 parts of Astragalus, 10 parts of Forsythia, 10 parts of Polygonatum, 10 parts of Isatis, 10 parts of Bupleurum, 5 parts of Panax notoginseng, 5 parts of Creeping Herb, 5 parts of Salvia miltiorrhiza, 10 parts of Licorice after drying to become fine powder with a particle size of less than 10μm, then put the fine powder into a blender and mix and stir for 30 minutes to make it thoroughly mixed), 12% of fish meal, 20% of snail meat, 15% of flour, 20% of soybean meal, 16% of corn flour, 5% of squid paste, 8% of fish oil and soybean oil, 1.5% of monocalcium phosphate, 0.1% of multivitamins and 0.4% of multiminerals, heat to above 85℃ with high-temperature steam for 30 minutes, granulate, cool and bag for later use.

[0069] The feed formula of the control group was as follows: 2% second-grade flour, 12% fish meal, 20% snail meat, 15% flour, 20% soybean meal, 16% corn meal, 5% squid paste, 8% fish oil and soybean oil, 1.5% monocalcium phosphate, 0.1% multivitamins, and 0.4% multiminerals.

[0070] 1.3 Experimental Grouping 200 crabs were randomly divided into 4 groups, 50 crabs in each group, and kept in mesh cages with a diameter × height = 50 cm × 45 cm ( Figure 2 ), 5 in each cage, 10 cages in each group. The first group was the control group, fed with commercial feed; the second group was fed with fermented feed; the third group was the Chinese herbal medicine pre-made feed group, fed with Chinese herbal medicine functional feed throughout the whole process; the fourth group was fed with interval feeding, fed with feed on the first day, and fed with Chinese herbal medicine functional feed on the 2nd-3rd day, and the cycle was repeated.

[0071] The breeding cages were placed in the adult crab pond of the breeding base, with a water depth of 50-60 cm. During the experiment, an appropriate amount of Elodea was placed in the net cages and replaced regularly. Feed was given once every evening, and the amount of feed given to each group was about 3% according to the feed coefficient. The uneaten feed was removed according to the remaining feed observed the next day. The experiment lasted from April 18 to May 14. The survival of river crabs was checked every day, the number of dead crabs was recorded, and the weight of dead crabs was weighed with an analytical balance. At the end of the experiment, the number of remaining crabs was counted and weighed. The survival rate = the number remaining after the end of the experiment.

[0072] The dried composite Chinese herbal medicine is crushed into fine powder with a particle size of less than 10 μm by using an ultra-fine pulverizer; like Figure 9-12As shown, the ultrafine pulverizer includes a body 1, a feed hopper 3 is provided on the top of the body 1, a crushing roller group 5 is installed at the bottom of the feed hopper 3, a refining mechanism 2 is provided at the bottom of the crushing roller group 5, and a containing mechanism 4 is provided inside the body 1, the crushing roller group 5 can be used to preliminarily crush the composite Chinese herbal medicine, and then the refining mechanism 2 can be used to crush and refine the crushed composite Chinese herbal medicine for multiple times, thereby effectively improving the crushing effect;The refinement mechanism 2 includes a first guide plate 201, a second guide plate 202 and a third guide plate 203. The surfaces of the first guide plate 201, the second guide plate 202 and the third guide plate 203 are all provided with a ceramic coating, so that the surfaces of the first guide plate 201, the second guide plate 202 and the third guide plate 203 are smoother, and feed residue is prevented, so that the feed can fall fully. The first guide plate 201, the second guide plate 202 and the third guide plate 203 are all fixedly installed on the machine body 1, and the first guide plate 201, the second guide plate 202 and the third guide plate 203 are all tiltedly arranged on the inner wall of the machine body 1, so that the first guide plate 201, the second guide plate 202 and the third guide plate 203 can be used to 2 and the third guide plate 203 play a guiding role for the feed, and two thinning knife rollers 204 are arranged between the first guide plate 201 and the second guide plate 202 and between the second guide plate 202 and the third guide plate 203. The ends of the two thinning knife rollers 204 are fixedly connected with support shafts 205, and the two support shafts 205 are fixedly connected with transmission gears 206. The two thinning knife rollers 204 are connected by transmission gears 206. The thinning knife rollers 204 are rotatably connected to the inner wall of the body 1 through the support shaft 205, so that the two thinning knife rollers 204 can rotate in opposite directions. A driving motor is fixedly connected to the end of one support shaft 205, and can be driven by the driving motor. The machine drives two thinning rollers 204 to rotate at the same time, so that the feed can be finely crushed. A fixed seat 207 is provided on one side of the two thinning rollers 204, and the fixed seat 207 is fixedly connected to the inner wall of the machine body 1. The two fixed seats 207 are respectively arranged at the bottom of the first guide plate 201 and the second guide plate 202, so that the two fixed seats 207 can vibrate the first guide plate 201 and the second guide plate 202, so that the feed on them can fall fully. The fixed seat 207 is internally rotatably connected with a rotating shaft 208, and the rotating shaft 208 and the end of a support shaft 205 are fixedly connected with a synchronous wheel 209, and a synchronous belt 210 is sleeved between the two synchronous wheels 209. A plurality of cams 211 are fixedly connected to the shaft 208, a stand 212 is provided on the top of the cam 211, a push rod 213 is slidably connected inside the stand 212, the stand 212 is fixedly mounted on the top of the fixed seat 207, the bottom end of the push rod 213 is in contact with the cam 211, so that the cam 211 can drive the push rod 213 to move, a limit block 215 is fixedly connected to the push rod 213, a return spring 214 is provided between the limit block 215 and the stand 212, one end of the return spring 214 is fixedly connected to the bottom surface of the stand 212, and the other end of the return spring 214 is fixedly connected to the top surface of the limit block 215, so that the return spring 214 can reset the push rod 213;The containing mechanism 4 comprises a containing box 401, a containing cavity 403 is provided inside the containing box 401, the containing box 401 is plugged with the machine body 1, a handle 402 is fixedly connected to the front surface of the containing box 401, and an adsorption magnet 404 is fixedly connected to the rear surface of the containing box 401. The containing box 401 can be inserted into the machine body 1 to receive the fully crushed and refined composite Chinese herbal medicine, and the adsorption magnet 404 is used to fix the compound Chinese herbal medicine by magnetic attraction, thereby improving the stability of use. ;

[0073] During operation, the dried composite Chinese herbal medicine is firstly crushed by the crushing roller group 5, and then falls along the first guide plate 201, the second guide plate 202 and the third guide plate 203 in sequence. During the falling process, the driving motor drives the refining roller 204 to rotate to refine and crush the feed. At the same time, the rotation of the refining roller 204 drives the rotating shaft 208 to rotate through the synchronous wheel 209 and the synchronous belt 210, and the rotation of the rotating shaft 208 drives the cam 211 to rotate. When the cam 211 rotates to lift the push rod 213 to rise, the return spring 214 is compressed. When the cam 211 and the push rod 213 are gradually separated, the return spring 214 recovers its deformation and pushes the push rod 213 to fall, and the continuous cycle drives the push rod 213 to move up and down, knocking the first guide plate 201 and the second guide plate 202 to produce vibration, so that the composite Chinese herbal medicine can fall fully to avoid residue, so that multiple crushing and refining can be carried out, effectively improving the crushing effect.

[0074] The above are only preferred specific implementation modes of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical solutions and inventive concepts of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A composite Chinese herbal medicine, characterized in that: The composite Chinese herbal medicine is prepared from the following components in parts by weight: 20-30 parts of Artemisia annua, 10-15 parts of Astragalus, 10-15 parts of Isatis indigotica, 5-10 parts of Bupleurum, 5-10 parts of Polygonatum, 5-10 parts of Panax notoginseng, 5-10 parts of Forsythia, 5-8 parts of Creeping Herb, 5-10 parts of Salvia miltiorrhiza, and 10-15 parts of Licorice.

2. A prefabricated feed for aquatic animals, characterized in that: The aquatic animal prefabricated feed contains the compound Chinese herbal medicine according to claim 1; when preparing the feed, the compound Chinese herbal medicine is directly added to the feed raw materials and processed together to prepare the feed.

3. The prefabricated feed for aquatic animals according to claim 2, characterized in that: Before making the feed, the compound Chinese herbal medicine is dried and then ultra-finely ground into fine powder with a particle size of less than 10 μm, and then the fine powder is put into a blender and mixed for 30 minutes to be fully mixed.

4. A prefabricated feed for aquatic animals, characterized in that: The raw material ratio of the prefabricated feed is: 5% Chinese medicine formula package, 11% fish meal, 12% cottonseed meal, 5% meat and bone meal, 14% soybean meal, 12% vegetable cake, 16% flour, 16.5% bran, 4% vegetable oil, 4% minerals, and 0.5% squid paste; The Chinese medicine formula package is made of the composite Chinese herbal medicine according to claim 1.

5. The aquatic animal prefabricated feed according to claim 3, characterized in that: The preparation of the Chinese medicine formula package is as follows: the compound Chinese herbal medicine is dried and then ultra-finely ground into fine powder with a particle size of less than 10 μm, and then the fine powder is put into a blender and mixed for 30 minutes to be fully mixed.

6. The prepared feed for aquatic animals according to claim 3, characterized in that: The preparation of the prefabricated feed is as follows: the raw materials are mixed evenly, heated to above 85° C. with high-temperature steam for 30 minutes, pelletized, cooled, and bagged.

7. A prefabricated feed for aquatic animals according to claim 2 or 5, characterized in that: The dried composite Chinese herbal medicine is crushed into fine powder with a particle size of less than 10 μm by using an ultra-fine pulverizer; The ultrafine pulverizer comprises a body, a feed hopper is provided on the top of the body, a crushing roller group is installed at the bottom of the feed hopper, a refining mechanism is provided at the bottom of the crushing roller group, and a containing mechanism is provided inside the body; the refining mechanism comprises a first guide plate, a second guide plate and a third guide plate, two refining knife rollers are provided between the first guide plate and the second guide plate and between the second guide plate and the third guide plate, the ends of the two refining knife rollers are fixedly connected to a support shaft, the two support shafts are fixedly connected to a transmission gear, one end of the support shaft is fixedly connected to a driving motor, a fixed seat is provided on one side of the two refining knife rollers, a rotating shaft is rotatably connected inside the fixed seat, the rotating shaft and an end of the support shaft are fixedly connected to a synchronous wheel, a synchronous belt is sleeved between the two synchronous wheels, and a plurality of A cam, a vertical frame is provided on the top of the cam, a push rod is slidably connected inside the vertical frame, a limit block is fixedly connected to the push rod, and a return spring is provided between the limit block and the vertical frame; the first guide plate, the second guide plate and the third guide plate are all fixedly mounted on the machine body, and the first guide plate, the second guide plate and the third guide plate are all inclinedly arranged on the inner wall of the machine body; the two refinement knife rollers are connected by transmission gears, and the refinement knife rollers are rotatably connected to the inner wall of the machine body through a support shaft; the fixed seat is fixedly connected to the inner wall of the machine body, and the two fixed seats are respectively arranged at the bottom of the first guide plate and the second guide plate; the vertical frame is fixedly mounted on the top of the fixed seat, and the bottom end of the push rod is in contact with the cam; one end of the return spring is fixedly connected to the bottom surface of the vertical frame, and the other end of the return spring is fixedly connected to the top surface of the limit block.

8. The prefabricated feed for aquatic animals according to claim 7, characterized in that: The containing mechanism comprises a containing box, a containing cavity is provided inside the containing box, and the containing box is plugged into the machine body; a handle is fixedly connected to the front surface of the containing box, and an adsorption magnet is fixedly connected to the rear surface of the containing box.

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