A plant-derived composite preparation for controlling sexual precocity of macrobrachium nipponense, a preparation method and a use method

Abstract

This invention belongs to the field of aquaculture technology, specifically relating to a plant-derived compound preparation for controlling precocious puberty in giant freshwater prawns, its preparation method, and its application method. The plant-derived compound preparation comprises the following raw materials in parts by weight: 1.5–3.0 parts gardenia extract, 0.5–1.2 parts quince extract, 0.5–1.0 parts mulberry leaf extract, 0.5–1.2 parts lily extract, 1.0–2.5 parts dried ginger extract, and 90–98 parts vitamin B complex powder. The plant-derived compound preparation is added to the feed of giant freshwater prawns at specific growth stages and simultaneously sprinkled into the aquaculture water. Through surface penetration and absorption, it regulates and improves hepatopancreatic function, enhances nutrient digestion and conversion, promotes growth, and reduces the probability of precocious puberty, thus replacing existing control measures. The technical solution of this invention has the characteristics of good and stable control effect, small dosage, convenient use, and high safety, ensuring the size and yield of marketable prawns, thereby achieving the goal of rationally controlling the density of giant freshwater prawns in ponds and improving aquaculture efficiency.

Description

Technical Field

[0001] This invention belongs to the field of aquaculture technology, specifically relating to a plant-derived compound preparation for controlling precocious sexual maturity in Japanese giant freshwater prawns, its preparation method, and its application method. Background Technology

[0002] Japanese prawn (Macrobrachium nipponense), also known as freshwater prawn or river prawn, is a famous and high-quality aquatic product in my country. Its delicate texture and delicious taste make it a favorite among people in the lower reaches of the Yangtze River. In 2022, my country's farmed Japanese prawn production reached 226,300 tons, with Jiangsu Province accounting for 104,000 tons, or 45.93% of the national total, ranking first in the country. Japanese prawn farming has become one of the leading industries in Jiangsu Province's freshwater fisheries. However, with the continuous expansion of Japanese prawn farming and the rapid development of the industry, many problems have emerged, such as genetic degradation, increased diseases, and a lack of diversified farming methods. In particular, early sexual maturity has led to low marketability, declining quality, and weak market competitiveness of Japanese prawns, seriously affecting the healthy development of Japanese prawn farming.

[0003] Precocious sexual maturity is one of the most prominent problems in Japanese giant freshwater prawn (MGR) farming. Prawn larvae bred that year, after 45-50 days of rearing, reach a body length of 3-4 cm and are ready to carry eggs and reproduce. Early sexual maturity in Japanese giant freshwater prawns leads to slow growth, with nutrients accumulating for gonadal development, resulting in generally smaller individuals, smaller marketable size, and a lower marketable rate (averaging only about 50%), directly impacting economic benefits. Precocious sexual maturity shortens the growth period of Japanese giant freshwater prawns, leading to excessive breeding of larvae in autumn and a rapid, uncontrollable increase in prawn density in the ponds. Competition between larvae and adults for food, oxygen, and space in the ponds results in uneven individual size, reduced marketability, and further impacts yield. Therefore, adopting appropriate measures to prevent premature sexual maturity in Japanese giant freshwater prawns is extremely important and urgent.

[0004] Currently, preventative measures in production mainly involve purchasing high-quality seedlings and stocking them in full at once, avoiding high-temperature periods for cultivation, planting aquatic plants in ponds to reduce light exposure and water temperature, and reducing the amount of high-protein feed. While these measures have some effect, the overall results are unsatisfactory. These measures primarily aim to influence the physiological functions of the Japanese giant prawn by altering its external living environment, but they can also lead to problems such as low pond water utilization, slower growth rates, longer production cycles, and reduced farming efficiency.

[0005] Therefore, under this premise, the control of precocious puberty through external penetration or internal administration has attracted attention, and it is on this basis that the technical solution of this invention is proposed. Summary of the Invention

[0006] To address the problems existing in the prior art, this invention provides a plant-derived compound preparation for controlling precocious puberty in giant freshwater prawns, its preparation method, and its application method. The plant-derived compound preparation is added to the feed of giant freshwater prawns at specific growth stages and simultaneously sprayed into the aquaculture water. Through surface penetration and absorption, it regulates and improves hepatopancreatic function, enhances nutrient digestion and conversion, promotes growth, and reduces the probability of precocious puberty, thus replacing existing control measures. The technical solution of this invention features good and stable control effects, small dosage, convenient use, and high safety. It can ensure the size and yield of marketable prawns, thereby achieving the goal of rationally controlling the density of giant freshwater prawns in ponds and improving aquaculture efficiency.

[0007] The present invention provides a plant-derived compound preparation for controlling precocious puberty in Japanese freshwater prawns, comprising the following raw materials in parts by weight: 1.5-3.0 parts of gardenia extract, 0.5-1.2 parts of malachite extract, 0.5-1.0 parts of mulberry leaf extract, 0.5-1.2 parts of lily extract, 1.0-2.5 parts of dried ginger extract, and 90-98 parts of vitamin B complex powder.

[0008] To facilitate understanding of this invention, the functions of the raw materials used in this invention are explained:

[0009] Gardenia extract: This is an extract from the fruit of *Gardenia jasminoides*, a plant in the Rubiaceae family. It is a reddish-brown powder containing chemical components such as geniposide, hydroxyisogeniposide, geniposide, neogeniposide, geniposide acid, geniposide flavonoids, saffron glycoside, saffron acid, and methyl paederin. Gardenia is an evergreen shrub, cultivated in most parts of China. Wild resources exist in southern regions, generally growing on hillsides and roadsides, mainly distributed in Jiangxi, Hunan, Zhejiang, Fujian, and Sichuan provinces. Gardenia is cold in nature and bitter in taste. It has the effects of clearing heat and detoxifying, reducing swelling and relieving pain, purging fire and relieving irritability, cooling blood and promoting diuresis and detoxification, protecting the liver and gallbladder, and calming and stabilizing blood pressure. No adverse reactions are observed after its use.

[0010] *Palatinate* extract: Extracted from the dried branches and leaves of *Palatinate*, a plant in the Simaroubaceae family, it is a brownish-yellow powder containing quassinolide A–N, azadirachtin C–G, quassinolide hemiacetal C–G, quassinolide ketone, methyl quassinolide ketone, I-hydroxymethyl-β-carboline, and other chemical components. *Palatinate* is a deciduous tree that grows in mixed forests at altitudes of 1400–2400 meters, mainly distributed in Liaoning, Hebei, Shanxi, Shaanxi, Gansu, Henan, Hubei, Hunan, and Jiangxi provinces. Its branches and leaves are generally harvested in summer and autumn and dried to produce medicinal materials. *Palatinate* is bitter and cold in nature, possessing properties of clearing heat and detoxifying, dispelling wind and dampness, anti-inflammatory and antibacterial, and insecticidal effects. However, it is slightly toxic, and dosage should be carefully monitored.

[0011] Mulberry leaf extract: A yellowish-brown powder obtained by processing mulberry leaf powder from the first to third new leaves on mulberry branches during the late spring silkworm rearing period or before the first frost, through enzyme-water extraction with heating and spray drying. It contains various physiologically active substances such as mulberry leaf flavonoids, mulberry leaf polyphenols, mulberry leaf polysaccharides, 1-deoxynojirimycin, and γ-aminobutyric acid. The mulberry tree is a deciduous tree or shrub belonging to the genus *Morus* of the family Moraceae. It is distributed in Guangxi, Yunnan, Hubei, and other regions, especially in the middle and lower reaches of the Yangtze River and the Sichuan Basin, where it grows in fields, along roadsides, or is planted in mountainous areas. Mulberry leaves are bitter and sweet in taste, and cold in nature. They have the effects of dispersing wind-heat, clearing the lungs and moistening dryness, calming the liver and improving eyesight, cooling the blood and stopping bleeding. They can also lower blood sugar, blood lipids, and blood pressure, and have antioxidant and antimicrobial properties.

[0012] Lily extract: Extracted from the underground fleshy bulb of the lily (Lilium brownii), a plant in the Liliaceae family, it is a brownish-yellow fine powder containing various active substances, such as phospholipids, phenolic glycerides, steroidal glycosides, steroidal alkaloids, proteins, and amino acids. Lily is a perennial herbaceous bulbous plant, mainly distributed in Hebei, Shanxi, and Henan provinces. It grows in grassy slopes, under sparse forests, beside mountain gullies, field edges, or near villages at altitudes of 300–920 meters, demonstrating strong adaptability. Lily has a sweet taste and slightly cold properties, possessing anti-inflammatory, antioxidant, antibacterial, hypoglycemic, hypolipidemic, sedative, yin-nourishing and lung-moistening, tonic, and anti-cancer effects.

[0013] Dried ginger extract: Extracted from the dried rhizome of ginger (Zingiber officinale), a plant in the ginger family (Zingiberaceae), it is a pale yellow powder containing volatile oils and gingerol, such as gingerol, ginger glycol, gingerol, gingerone, and ginger dione, which are physiologically active substances. Ginger is a perennial herbaceous plant belonging to the genus Zingiber in the ginger family (Zingiberaceae), with yellowish-green flowers and a rhizome with a pungent aroma. Ginger is widely cultivated in central, southeastern, and southwestern China, and is also commonly cultivated in tropical Asia. Ginger combines nutrition and health benefits, and has been regarded by medical practitioners since ancient times as a health food with medicinal properties. Ginger has a pungent taste and slightly warm nature, and has multiple functions such as relieving exterior syndromes and dispelling cold, warming the middle jiao and stopping vomiting, warming the lungs and stopping cough, detoxifying and removing dampness, and accelerating blood circulation. It can also promote appetite, warm the middle jiao and relieve pain, and has antibacterial and detoxifying effects.

[0014] Jiawei powder: a complex made from various minerals, inorganic salts, modified starch, and other excipients through a special process. It is characterized by its non-hygroscopic nature, non-caking properties, good water solubility, excellent flowability, uniform color, and stable properties, and is commonly used as a carrier. For soluble drugs using it as a carrier, the solubility and dissolution rate of the main components increase, which is beneficial for the rapid absorption and distribution of the drug in the body, greatly improving the bioavailability and efficacy of the drug. Many manufacturers produce Jiawei powder (some also call it Jiayi powder). The Jiawei powder of this invention is selected from Xingtai Rongda Biotechnology Co., Ltd.

[0015] In summary, the plant-derived compound preparation provided by this invention uses gardenia extract, nanmu extract, mulberry leaf extract, lily extract, and ginger extract as its main components, supplemented with mineral carriers and vitamin powder in a scientifically and rationally proportioned weight ratio. Among them:

[0016] 1) Gardenia extract is rich in natural active substances such as geniposide, geniposide, geniposide acid, and genistein. It can enter organs such as the liver through skin penetration and food absorption, clearing internal heat and reducing liver fire, repairing easily damaged liver cells, removing various metabolic oxidation products and free radicals, balancing the body's endocrine system, regulating the function of the hypothalamus-pituitary-gonadal axis, inhibiting the secretion of hormones such as cortisol and estradiol, and controlling precocious puberty; at the same time, it promotes bile excretion, improves the absorption of various nutrients in feed, promotes shrimp growth, and improves the size of marketable shrimp.

[0017] 2) The extract of *Quercus mongolica* contains natural chemical components such as quassinolide, quassinolide hemiacetal, quassinolide ketone, and azadirachtin, which have the effects of clearing heat and detoxifying. Its effect is enhanced when combined with gardenia extract. At the same time, *Quercus mongolica* extract has anti-inflammatory, antibacterial, and insecticidal effects, which can reduce the use of some antibiotics and chemical drugs in the aquaculture process and reduce the intake of environmental estrogens by shrimp during the farming process.

[0018] 3) Mulberry leaf extract contains a variety of physiologically active substances such as mulberry leaf flavonoids, mulberry leaf polyphenols, mulberry leaf polysaccharides, 1-deoxynojirimycin, and γ-aminobutyric acid. The combined effect of these substances can inhibit the conversion of sugar in the body, reduce fat accumulation, and reduce the conversion of lipids into estrogen, which can further control the increase of sex hormones and control precocious puberty to a certain extent. It can also promote the secretion of growth hormone and promote the growth of shrimp.

[0019] 4) Lily extract contains phospholipids, phenolic acid glycerides, steroidal glycosides, steroidal alkaloids, proteins, amino acids and other active substances. It has anti-inflammatory and antibacterial effects, and has a calming and sedative effect. It can improve shrimp immunity and promote shrimp growth.

[0020] 5) Dried ginger extract contains volatile oils and gingerol, such as gingerol, gingerene, gingerol, and gingerone, which are physiologically active substances that can detoxify, remove dampness, accelerate blood circulation, and promote excretion. Dried ginger, when combined with the above-mentioned substances, has a synergistic effect. On the one hand, it can promote appetite, accelerate the absorption, utilization, and transformation of these substances, and accelerate the excretion of metabolic products; on the other hand, dried ginger's pungent and warm nature can counteract the excessive effects of the aforementioned cold substances, achieving the goal of controlling precocious puberty without affecting normal development.

[0021] 6) The added vitamin powder is an inorganic salt carrier with good dispersibility, which is conducive to the mixing of plant extracts and prevents clumping after mixing. At the same time, it supplements inorganic salts and minerals, increases the nutrition of shrimp, and facilitates the conversion and utilization of plant extracts.

[0022] The raw materials described in this invention work synergistically to control precocious puberty in Japanese freshwater prawns, improve prawn size, and increase yield. This invention integrates multiple functions such as clearing heat and reducing lipids, protecting the liver and promoting digestion, inhibiting precocious puberty, improving prawn size, and increasing yield, resulting in significant and stable control effects.

[0023] Preferably, the plant-derived compound preparation for controlling precocious puberty in Japanese freshwater prawns comprises the following raw materials in parts by weight: 2.0 parts gardenia extract, 0.8 parts madder extract, 0.8 parts mulberry leaf extract, 0.7 parts lily extract, 1.5 parts dried ginger extract, and 94.2 parts vitamin B1 powder.

[0024] Preferably, the gardenia extract is a water-soluble powder with a ratio of 20:1 and a fineness of 80-100 mesh.

[0025] And / or, the extract of the bitter tree is a water-soluble powder with a specification of 10:1 and a fineness of 100-120 mesh;

[0026] And / or, the mulberry leaf extract is a water-soluble powder with a specification of 10:1 and a fineness of 80-100 mesh;

[0027] And / or, the lily extract is a water-soluble powder with a ratio of 10:1 and a fineness of 80-100 mesh;

[0028] And / or, the ginger extract is a water-soluble powder with a ratio of 20:1 and a fineness of 100-120 mesh.

[0029] Preferably, the preparation method of the gardenia extract, the nanmu extract, or the dried ginger extract includes the following steps:

[0030] (1) Mix dried gardenia, dried bitterwood or dried ginger with water, stir well and heat to the first temperature and keep warm and stand;

[0031] (2) Then, the pH value was adjusted under stirring conditions, and a compound enzyme was added for enzymatic hydrolysis;

[0032] (3) After the enzymatic hydrolysis is completed, heat the system to the second temperature, keep it warm, then stop heating and stirring. After it cools down naturally, filter it, discard the filter residue, and obtain the filtrate.

[0033] (4) The filtrate is spray-dried to obtain the gardenia extract, the bitterwood extract or the dried ginger extract.

[0034] Preferably, in step (1), the weight ratio of the dried gardenia, dried bitterwood, or dried ginger to water is 8-15:55-80; the first temperature is 40-55℃; and the heat preservation and standing time is 60-120 min.

[0035] And / or, in step (2), the stirring rate is 100-200 r / min; the pH value is 5.5-7.5; the complex enzyme is a neutral protease and a cellulase, and the amount of the complex enzyme added is 0.5-1.0% of the total mass; the enzymatic hydrolysis time is 40-90 min; wherein, the cellulase activity is 50,000-100,000 U / g, and the neutral protease activity is 100,000-200,000 U / g (the same below);

[0036] And / or, in step (3), the second temperature is 85-95°C; the heat preservation time is 60-90 min.

[0037] Preferably, the preparation method of the mulberry leaf extract or the lily extract includes the following steps:

[0038] (S1) Mix fresh mulberry leaves or fresh lilies with water, stir well, heat to the first temperature and keep warm and stand;

[0039] (S2) Then, adjust the pH value under stirring conditions and add mulberry leaf compound enzyme or lily compound enzyme for enzymatic hydrolysis.

[0040] (S3) After the enzymatic hydrolysis is complete, heat the system to the second temperature, keep it at the temperature, then stop heating and stirring. After it cools down naturally, filter it, discard the filter residue, and obtain the filtrate.

[0041] (S4) The filtrate is spray-dried to obtain the mulberry leaf extract or the lily extract.

[0042] Preferably, in step (S1), the weight ratio of the fresh mulberry leaves or fresh lilies to water is 8-15:55-80; the first temperature is 40-55℃; and the heat preservation and standing time is 60-120 minutes.

[0043] And / or, in step (S2), the stirring rate is 100–200 r / min; the pH value is 5.5–7.5; the mulberry leaf complex enzyme is amylase and cellulase, and the lily complex enzyme is amylase, pectinase, and cellulase; the amount of the mulberry leaf complex enzyme or the lily complex enzyme added is 0.5–1.0% of the total mass; the enzymatic hydrolysis time is 40–90 min; wherein, the amylase activity is 100,000–200,000 U / g, and the pectinase activity is 25,000–50,000 U / g;

[0044] And / or, in step (S3), the second temperature is 85-95°C; the heat preservation time is 60-90 min.

[0045] Based on the same technical concept, another aspect of the present invention is to provide a method for preparing a plant-derived compound preparation for controlling precocious puberty in Japanese freshwater prawns, wherein the preparation method is as follows: all raw materials are mixed evenly.

[0046] Based on the same technical concept, another aspect of the present invention is to provide a method for using a plant-derived compound preparation for controlling precocious puberty in Japanese freshwater prawns, the method comprising the following steps:

[0047] (SS1) The plant-derived compound preparation is mixed with feed to obtain feed mix;

[0048] (SS2) When the Japanese giant prawn larvae are cultured in the pond for 20-26 days, the above-mentioned feed is added to the pond for the Japanese giant prawn larvae to take orally once a day for 5-8 days, and then stopped for 3-5 days.

[0049] (SS3) During the period of discontinuation, spray 100-200g of the plant-derived compound preparation per mu of water surface and per meter of water depth per day;

[0050] (SS4) One cycle is from the first day of adding the feed in step (SS2) to the last day of spraying the plant-derived compound preparation in step (SS3). Repeat 2 to 3 cycles.

[0051] Preferably, in step (SS1), the weight ratio of the plant-derived compound preparation to the feed is 0.3 to 0.6:100.

[0052] The beneficial effects of this invention are as follows:

[0053] 1. The plant-derived compound preparation of the present invention can effectively inhibit precocious puberty in Japanese giant prawns, promote growth, improve the size and uniformity of farmed prawns, and increase farming efficiency. Simultaneously, the plant-derived compound preparation can be directly added to feed or sprinkled into the aquaculture water, making it convenient to use, requiring only a small dosage, and exhibiting stable effects. It inhibits precocious puberty in Japanese giant prawns from both internal and external perspectives, while also promoting digestion, reducing disease incidence, and decreasing the use of antibiotics and other drugs, ultimately achieving the goal of increasing farming yield, quality, and efficiency. Furthermore, the plant-derived compound preparation is easily degradable, has a short residual period, high safety, and does not pollute the environment or food, controlling precocious puberty in Japanese giant prawns while having no toxic effects on aquatic animals.

[0054] 2. The preparation method of the plant-derived compound preparation of the present invention only requires mixing the raw materials evenly. The process is simple, easy to control, and conducive to large-scale production. Detailed Implementation

[0055] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0056] Example 1

[0057] This embodiment provides a method for preparing a plant-derived compound preparation for controlling precocious puberty in Japanese freshwater prawns, the preparation method comprising the following steps:

[0058] (I) Preparation of Gardenia Extract:

[0059] (1) Place 8g of dried gardenia into a stainless steel covered container with a stirrer, add 55g of purified water, stir well, heat to 40℃ and keep warm for 60min.

[0060] (2) Then, adjust the pH of the system to 6.5 at 100 r / min, and add 0.5% of the total mass of the system of the complex enzyme (including neutral protease and cellulase, with neutral protease activity of 100,000 U / g and cellulase activity of 50,000 U / g) for enzymatic hydrolysis for 40 min.

[0061] (3) After the enzymatic hydrolysis is completed, heat the system temperature to 85℃, keep it warm for 60 minutes, then stop heating and stirring. After it cools down naturally, filter it through a 50-mesh filter cloth, discard the filter residue, and obtain the filtrate.

[0062] (4) The gardenia extract is obtained by spray drying the filtrate.

[0063] (II) Preparation of *Paspalum notatum* extract:

[0064] Compared to "Preparation of Gardenia Extract", simply replace "dried gardenia" with "dried bitterwood", and keep all other operations the same.

[0065] (III) Preparation of dried ginger extract:

[0066] Compared to "Preparation of Gardenia Extract", simply replace "dried gardenia" with "dried ginger"; all other operations remain the same.

[0067] (IV) Preparation of mulberry leaf extract:

[0068] Compared with "Preparation of Gardenia Extract", "dried gardenia" is replaced with "fresh mulberry leaves"; "complex enzyme" is replaced with "mulberry leaf complex enzyme", wherein: the mulberry leaf complex enzyme includes amylase and cellulase, with amylase activity of 100,000 U / g and cellulase activity of 50,000 U / g.

[0069] (V) Preparation of lily extract:

[0070] Compared with "Preparation of Gardenia Extract", "dried gardenia" is replaced with "fresh lily"; "complex enzyme" is replaced with "lily complex enzyme", wherein: the lily complex enzyme includes amylase, pectinase and cellulase, with amylase activity of 100,000 U / g, pectinase activity of 25,000 U / g and cellulase activity of 50,000 U / g.

[0071] (VI) Preparation of plant-derived compound formulations:

[0072] Mix 2g of gardenia extract, 0.8g of bitter melon extract, 0.8g of mulberry leaf extract, 0.7g of lily extract, 1.5g of dried ginger extract, and 94.2g of vitamin B complex powder at room temperature until they are evenly mixed. After mixing, dispense, package, and store the mixture to obtain the plant-derived compound preparation.

[0073] This embodiment also provides a method for using a plant-derived compound preparation for controlling precocious puberty in Japanese freshwater prawns, the method comprising the following steps:

[0074] (S1) Mix the plant-derived compound preparation with the feed (add 0.4 kg of plant-derived compound preparation per 100 kg of feed) to obtain the mixed feed;

[0075] (S2) When the Japanese giant prawn larvae are cultured in the pond for 22-24 days, the above-mentioned feed is added to the pond for the Japanese giant prawn larvae to take orally once a day for 5 consecutive days, and then stopped for 3 days.

[0076] (S3) During the period of discontinuation, spray 150g of the plant-derived compound preparation per mu of water surface and per meter of water depth per day;

[0077] (S4) One cycle is defined as the first day of adding the feed in step (S2) to the last day of spraying the plant-derived compound preparation in step (S3). Repeat this cycle 3 times.

[0078] Example 2

[0079] The difference between Example 2 and Example 1 lies only in step (VI); all other operations are the same. Step (VI) of this example is as follows:

[0080] Mix 1.5g gardenia extract, 1.2g bitter melon extract, 1.0g mulberry leaf extract, 1.2g lily extract, 2.0g dried ginger extract, and 93.1g vitamin powder at room temperature until homogeneous. After mixing, dispense, package, and store to obtain the plant-derived compound preparation.

[0081] Example 3

[0082] The difference between Example 3 and Example 1 lies only in step (VI); all other operations are the same. Step (VI) of this example is as follows:

[0083] Mix 3.0g gardenia extract, 0.5g bitter melon extract, 0.5g mulberry leaf extract, 0.5g lily extract, 2.5g dried ginger extract, and 93.0g vitamin powder at room temperature until homogeneous. After mixing, dispense, package, and store to obtain the plant-derived compound preparation.

[0084] Example 4

[0085] The difference between Example 4 and Example 1 lies only in step (VI); all other operations are the same. Step (VI) of this example is as follows:

[0086] Mix 1.5g gardenia extract, 0.5g bitter melon extract, 0.5g mulberry leaf extract, 0.5g lily extract, 1.0g dried ginger extract, and 96.0g vitamin powder at room temperature until homogeneous. After mixing, dispense, package, and store to obtain the plant-derived compound preparation.

[0087] Example 5

[0088] The difference between Example 5 and Example 1 lies only in step (VI); all other operations are the same. Step (VI) of this example is as follows:

[0089] 3.0g of gardenia extract, 1.2g of bitter melon extract, 1.0g of mulberry leaf extract, 1.2g of lily extract, 2.5g of dried ginger extract, and 91.1g of vitamin B complex powder were stirred at room temperature until they were evenly mixed. After mixing, the mixture was dispensed, packaged, and stored to obtain the plant-derived compound preparation.

[0090] Comparative Example 1

[0091] The difference from Example 1 is that the raw material composition of this comparative example does not contain plant-derived extracts, but only 100g of vitamin powder.

[0092] Comparative Example 2

[0093] The only difference from Example 1 is the absence of dried ginger extract (but the content of added vitamin B1 is more adaptable, the same below). The preparation method of this comparative plant-derived compound preparation is as follows:

[0094] Mix 2.0g of gardenia extract, 0.8g of bitter melon extract, 0.8g of mulberry leaf extract, 0.7g of lily extract, and 95.7g of vitamin B complex powder at room temperature until they are evenly mixed. After mixing, dispense, package, and store the mixture to obtain the plant-derived compound preparation.

[0095] Comparative Example 3

[0096] The only difference from Example 1 is the absence of gardenia extract and nanmu extract (but the content of vitamin C powder is increased). The preparation method of this comparative plant-derived compound preparation is as follows:

[0097] Mix 0.8g of mulberry leaf extract, 0.7g of lily extract, 1.5g of dried ginger extract, and 97g of vitamin B complex powder at room temperature until they are evenly mixed. After mixing, dispense, package, and store the mixture to obtain the plant-derived compound preparation.

[0098] Comparative Example 4

[0099] The difference from Example 1 is that the plant-derived compound preparation was used 8 to 10 days later in Comparative Example 4 than in Example 1 after the shrimp larvae were introduced into the pond.

[0100] Comparative Example 5

[0101] The difference from Example 1 lies in the method of use. Example 1 uses a combination of oral administration and water spraying, while Comparative Example 5 uses only oral administration.

[0102] Verification Example

[0103] This invention is used for experimental research on the effect of controlling precocious puberty in Japanese freshwater prawns.

[0104] 1. Research locations: Experimental ponds A1-A6 for Japanese giant freshwater prawn farming in Lishui District, Nanjing, Jiangsu Province, and experimental ponds B1-B6 for Japanese giant freshwater prawn farming in Wujiang District, Suzhou, Jiangsu Province. The source of the prawn larvae, the stocking time and density, the feed used, and the management measures were consistent in each region.

[0105] 2. Method of use: The plant-derived compound preparations of Example 1 and Comparative Examples 1-3 of this invention shall be administered orally to shrimp fry 22-24 days after they are introduced into the pond (late May). Add 0.4 kg per 100 kg of feed for 5 consecutive days, then stop for 3 days. During the period of stopping the oral administration, sprinkle 150 g per mu of water surface per meter of water depth per day. This is one cycle, and repeat for 3 consecutive cycles.

[0106] Comparative Example 4 samples were tested 30–32 days after the shrimp larvae were introduced into the pond for culture, using the same method as in Example 1.

[0107] Comparative Example 5 samples were given the shrimp larvae orally 22-24 days after they were introduced into the pond. 0.4 kg of the larvae was added to every 100 kg of feed and administered for 5 consecutive days, followed by a 3-day break. This constituted one cycle, which was repeated for 3 consecutive cycles.

[0108] 3. Indicator Detection Methods: 60 days after the shrimp larvae were introduced into the pond, 30 shrimp were randomly selected from each experimental group. The shrimp body length (from the front of the shrimp's eye to the end of the tail with muscle) was measured using calipers, and the presence of eggs (whether there were roe on the shrimp's abdomen) was observed. Then, hemolymph was extracted to determine cortisol content, and hepatopancreas was collected to determine estradiol content. The hemolymph extraction method was as follows: the shrimp bodies were dried with absorbent paper, surface-sterilized with 75% alcohol, and then extracted from the pericardial cavity using a sterile syringe (1 mL, fitted with a No. 5 needle). Approximately 0.1 mL was extracted from each shrimp. The hemolymph from all shrimp was mixed, incubated at 4℃ for 12 hours, and then centrifuged at 4℃ and 3500 rpm for 10 minutes to prepare the supernatant for analysis. After the liver and pancreas were removed and combined, they were rinsed in physiological saline at 4℃, blotted dry with filter paper, and 2g was placed in a 50mL beaker. 12mL of homogenizing medium (containing 0.01mol / L sucrose, 0.01mol / L Tris-HCl, 0.0001mol / L Na2EDTA, 0.14mol / L NaCl, pH 7.4) at 4℃ was added to the beaker. The mixture was then poured into a homogenizer, and the beaker was rinsed with 6mL of the same homogenizing medium, which was then added to the homogenizer. The homogenizer was thoroughly homogenized in an ice-water bath to prepare a 10% (w / w) hepatopancreatic tissue homogenate. Cortisol and estradiol levels were detected using ELISA kits provided by Shanghai Enzyme-Link Biotechnology Co., Ltd. and Nanjing Jiancheng Biotechnology Co., Ltd., respectively, according to the kit instructions. During the 100-day rearing period, 50 shrimp were randomly selected from each group, weighed, and their body length (from the front of the shrimp's eyes to the end of the shrimp's tail with muscle) was measured using calipers to calculate the proportion of small-sized shrimp of about 3cm.

[0109] 4. Effects of Use: In ponds where the product of this invention was not used or was used improperly, Japanese freshwater prawns exhibited high levels of cortisol in their hemolymph, high levels of estradiol in their hepatopancreas, and precocious sexual maturity, resulting in inconsistent body size and smaller weight. After the scientific and reasonable use of the product of this invention, the levels of cortisol in the hemolymph and estradiol in the hepatopancreas of Japanese freshwater prawns significantly decreased, precocious sexual maturity was significantly reduced, body size became more uniform, and weight increased. Specific effects are shown in Table 1.

[0110] Table 1

[0111]

[0112]

[0113] Note: 1) Experimental method*: Start administering the feed orally 22, 24, 30 or 32 days after the shrimp larvae are introduced into the pond. Add 0.4 kg per 100 kg of feed for 5 consecutive days, then stop for 3 days. During the period of stopping the oral administration, sprinkle 150 g per mu of water surface per meter of water depth per day. This is one cycle, and repeat for 3 consecutive cycles.

[0114] 2) Experimental method**: 22 or 24 days after the shrimp larvae were introduced into the pond, the 0.4 kg of the 0.4 kg ...

[0115] Table 1 (Continued from Table 1)

[0116]

[0117]

[0118] Table 1 (Continued from Table 2)

[0119]

[0120] As can be seen from Table 1:

[0121] Using a sample containing only the carrier (Comparative Example 1), Japanese giant freshwater prawns exhibited higher cortisol levels in their hemolymph and estradiol levels in their hepatopancreas at 60 days of culture. These prawns were smaller in size and showed signs of precocious puberty, with a higher proportion of berried prawns. At 100 days, the prawns were of inconsistent size and had lower yields. After using a compound preparation containing plant-derived components, the precocious puberty of Japanese giant freshwater prawns in each experimental pond was improved to varying degrees under different application methods. Example 1 showed the best control and improvement effect, indicating that its formulation and application method were the most reasonable, and its effects on controlling precocious puberty and promoting growth were significant.

[0122] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A plant-derived composite preparation for controlling sexual precocity of Macrobrachium nipponense, characterized in that, The application relates to a preparation method of a Chinese medicine composition, and belongs to the technical field of Chinese medicine. The preparation method of the gardenia extract, the picrasma quassioides extract or the dried ginger extract comprises the following steps: (1) mixing dried gardenia, dried picrasma quassioides or dried dried ginger with water, stirring uniformly, heating to a first temperature and standing after heat preservation; (2) then adjusting the pH value under stirring and adding a compound enzyme for enzyme hydrolysis; (3) after the enzyme hydrolysis is completed, the system temperature is heated to a second temperature, heat preservation is carried out, then heating and stirring are stopped, natural cooling is carried out, filtration is carried out, the filter residue is discarded, and a filtrate is obtained; (4) the filtrate is spray dried to obtain the gardenia extract, the picrasma quassioides extract or the dried ginger extract; wherein: In step (1), the weight ratio of the dried gardenia, the dried picrasma quassioides or the dried dried ginger to water is 8-15:55-80; the first temperature is 40-55 DEG C; and the standing time after heat preservation is 60-120 min; And / or, in step (2), the stirring rate is 100-200 r / min; the pH value is 5.5-7.5; the compound enzyme is neutral protease and cellulase; the addition amount of the compound enzyme is 0.5-1.0% of the total mass; and the enzyme hydrolysis time is 40-90 min; And / or, in step (3), the second temperature is 85-95 DEG C; and the heat preservation time is 60-90 min; The preparation method of the mulberry leaf extract or the lily extract comprises the following steps: (S1) mixing fresh mulberry leaves or fresh lily with water, stirring uniformly, heating to a first temperature and standing after heat preservation; (S2) then adjusting the pH value under stirring and adding a mulberry compound enzyme or a lily compound enzyme for enzyme hydrolysis; (S3) after the enzyme hydrolysis is completed, the system temperature is heated to a second temperature, heat preservation is carried out, then heating and stirring are stopped, natural cooling is carried out, filtration is carried out, the filter residue is discarded, and a filtrate is obtained; (S4) the filtrate is spray dried to obtain the mulberry leaf extract or the lily extract; wherein: In step (S1), the weight ratio of the fresh mulberry leaves or the fresh lily to water is 8-15:55-80; the first temperature is 40-55 DEG C; and the standing time after heat preservation is 60-120 min; And / or, in step (S2), the stirring rate is 100-200 r / min; the pH value is 5.5-7.5; the mulberry compound enzyme is amylase and cellulase, and the lily compound enzyme is amylase, pectinase and cellulase; the addition amount of the mulberry compound enzyme or the lily compound enzyme is 0.5-1.0% of the total mass; and the enzyme hydrolysis time is 40-90 min; And / or, in step (S3), the second temperature is 85-95 DEG C; and the heat preservation time is 60-90 min.

2. The plant-derived compound preparation for controlling the sexual precocity of Macrobrachium nipponense according to claim 1, characterized in that, The gardenia extract, the quassia extract, the mulberry leaf extract, the lily extract and the dried ginger extract are in the form of water-soluble powder.

3. The plant-derived composite preparation for controlling the sexual precocity of Macrobrachium nipponense according to claim 1 or 2, characterized in that, The gardenia extract is in the form of water-soluble powder with a specification of 20:1 and a fineness of 80-100 mesh. The quassia extract is in the form of water-soluble powder with a specification of 10:1 and a fineness of 100-120 mesh. The mulberry leaf extract is in the form of water-soluble powder with a specification of 10:1 and a fineness of 80-100 mesh. The lily extract is in the form of water-soluble powder with a specification of 10:1 and a fineness of 80-100 mesh. The dried ginger extract is in the form of water-soluble powder with a specification of 20:1 and a fineness of 100-120 mesh.

4. The method for preparing the plant-derived complex preparation for controlling the sexual precocity of Macrobrachium nipponense according to any one of claims 1 to 3, characterized in that, The preparation method is to mix all the raw materials uniformly.

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