Special nutrient feed for female turtles in egg laying period

Abstract

This invention relates to the technical field of aquaculture feed, specifically a special nutritional feed for laying female turtles, comprising the following ingredients by weight: 45-60% Cordyceps militaris mycelium, 20-25% wheat bran, 12-15% corn flour, 5-10% tomato powder, and 3-5% oyster meat powder. The Cordyceps militaris mycelium is obtained by mixing a culture medium containing 35-50% soybean meal, 20-25% sugarcane, 15-20% Leonurus japonicus, 8-10% oyster shell powder, 5-7% soybean flour, and 2-3% silkworm pupa powder with lotus root sauce, placing it evenly in a cultivation box, sterilizing it, and then inoculating and cultivating it. The inoculum is isolated from the fruiting bodies of Cordyceps militaris and then cultivated into a liquid inoculum. This special nutritional feed for laying female turtles has a cleverly formulated ingredient ratio, achieving multiple goals such as enhancing immunity, supplementing ovulation nutrition, improving reproductive performance, preventing postpartum inflammation, and shortening the recovery period. Furthermore, this feed is highly safe and free of drug residues.

Description

Technical Field

[0001] This invention relates to the technical field of aquaculture feed, specifically a nutritional feed for laying female turtles. Background Technology

[0002] Turtles are symbols of longevity. They not only have high nutritional and medicinal value but also possess ornamental value. In recent years, the price of turtles has been rising, but the number of wild turtles is declining sharply. Therefore, artificial turtle farming is becoming a new hot topic in aquaculture. To improve the economic benefits of turtle farming, it is essential to ensure the health of the female turtles and maximize their egg-laying capacity.

[0003] Currently, the feed given to laying female turtles is mostly general-purpose aquatic feed, or simply supplemented with high-protein ingredients such as fishmeal and soybean meal. However, the egg-laying period is the most metabolically active stage for female turtles. During this period, they not only need to ingest a large amount of nutrients to maintain their normal physiological functions, but also need to provide sufficient energy and nutrients for the formation and development of eggs. At the same time, the reproductive system of female turtles is prone to mechanical damage during egg-laying, and their postpartum immunity is significantly reduced, making them highly susceptible to postpartum inflammations such as endometritis and pelvic inflammatory disease. This leads to a prolonged postpartum recovery period, and in severe cases, it can affect subsequent reproductive performance or even cause the death of the female turtle. Feeding egg-laying female turtles with traditional feed has the following core defects: First, the nutritional ratio lacks specificity, failing to accurately supplement the specific nutrients (such as high-quality protein and immune-active substances) required for egg laying, resulting in low egg production and poor fertilization and hatching rates. Second, it does not consider the need for postpartum inflammation prevention in female turtles, as the feed does not contain ingredients with anti-inflammatory and repair functions, leading to a high incidence of postpartum inflammation and a long recovery period. Third, some feeds use antibiotics and other drugs to prevent inflammation, which can easily cause drug residues, affecting not only the quality of turtle products but also damaging the aquaculture ecosystem.

[0004] Research has revealed that there is currently no specific nutritional feed for female turtles that can simultaneously increase egg production, improve fertilization rates, and shorten the postpartum recovery period. Given these limitations, developing a specialized nutritional feed for laying female turtles that combines multiple functions—enhancing immunity, supplementing nutrients needed for egg production, and preventing postpartum inflammation—while also being free of drug residues and highly safe, is of significant practical importance for improving the reproductive performance of female turtles and ensuring the profitability of turtle farming. Summary of the Invention

[0005] To address the technical problems of insufficient targeted nutritional formulation and ineffective prevention of postpartum inflammation in existing egg-laying female turtle feed, this invention provides a special nutritional feed for egg-laying female turtles. This special nutritional feed for egg-laying female turtles uses a precise ratio of six functional ingredients: Cordyceps militaris mycelium, oyster meat powder, oyster shell powder, sugarcane, motherwort, and tomato, which are synergistically formulated with basic feed to achieve multiple goals such as enhancing immunity, supplementing egg-laying nutrition, improving reproductive performance, preventing postpartum inflammation, and shortening the recovery period. Moreover, this feed has low production cost, high safety, and no drug residues.

[0006] This invention achieves its objective through the following approach: A special nutritional feed for laying female turtles, comprising the following ingredients by weight percentage: 45-60% Cordyceps militaris mycelium, 20-25% wheat bran, 12-15% corn flour, 5-10% tomato powder, and 3-5% oyster meat powder; wherein the Cordyceps militaris mycelium is cultivated by the following method: (1) Collection and cultivation of strains: collect Cordyceps militaris fruiting bodies cultivated with silkworm pupae as parasites, isolate strains from them, and then cultivate them into liquid strains according to conventional microbial strain cultivation methods for later use.

[0007] (2) Take an appropriate amount of fresh lotus root and make it into a sauce. When making the sauce, the mass ratio of fresh lotus root to water is 2:(10-15). Then, the mass ratio of culture medium to lotus root sauce is 1:(1.3-1.5). Mix the culture medium and lotus root sauce evenly and place them in a cultivation box for sterilization. The culture medium is made by weight percentage as follows: 35-50% soybean meal, 20-25% sugarcane, 15-20% motherwort, 8-10% oyster shell powder, 5-7% soybean powder, and 2-3% silkworm pupa powder. The soybean meal, sugarcane, soybean powder, and silkworm pupa powder in the formula constitute a compound nitrogen source system. Soybean meal is a high-quality source of plant protein. Sugarcane is rich in natural sugars, dietary fiber, and various minerals, and is a high-quality natural carbon source. Silkworm pupa powder provides animal protein and essential amino acids. The combination of the above-mentioned soybean meal, sugarcane, soybean powder, and silkworm pupa powder helps to balance the amino acid composition and meet the nitrogen source requirements of mycelial growth. The addition of oyster shell powder primarily aims to provide abundant calcium and trace elements. These minerals are not only essential nutrients for mycelial growth but also remain in the mycelium after fermentation, enhancing the mineral nutritional value of the final feed product. Fresh lotus root is rich in starch, soluble sugars, plant protein, amino acids, phosphorus / potassium / magnesium minerals, and active substances such as lotus root polyphenols, polysaccharides, and flavonoids, making it a high-quality natural substrate for cultivating Cordyceps militaris. Lotus root starch provides a stable and sufficient carbon source for Cordyceps militaris mycelial growth. Lotus root is rich in phosphorus, potassium, magnesium, and calcium minerals. Combined with soybean meal, sugarcane, and silkworm pupa powder in the formula, it can completely replace the glucose, potassium dihydrogen phosphate, magnesium sulfate, and peptone that need to be added during traditional Cordyceps militaris cultivation. The polyphenols, polysaccharides, and flavonoids contained in lotus root can significantly promote the synthesis of cordycepin and cordyceps polysaccharides.

[0008] (3) Under sterile conditions, the above liquid inoculum is inoculated into the culture medium. First, it is cultured in the dark until the mycelium completely fills the culture medium, and then it is cultured in the light until the mycelium turns into a uniform orange-yellow color.

[0009] (4) Dry the mycelium that has completed color change at a constant temperature of 50°C until the moisture content of the material is less than 10%. After cooling, crush it to a particle size that is compatible with other raw materials and set aside for later use.

[0010] The Cordyceps militaris mycelium must meet the following weight percentage requirements: mycelium content ≥50%, crude protein content ≥20%, polysaccharide content ≥10%, moisture content ≤8%, and free from mold and odor.

[0011] Furthermore, the raw material for preparing the tomato powder is fresh tomatoes, and the preparation process is low-temperature drying followed by pulverization, meeting the following indicators: particle size 80-120 mesh, moisture content ≤6%.

[0012] Furthermore, the oyster meat powder is made from fresh oyster meat, and the preparation process is as follows: the fresh oyster meat is dried at a low temperature of 60-70℃ and then pulverized to a particle size of 80-120 mesh. The oyster meat powder must meet the following weight percentage indicators: crude protein content ≥50% and moisture content ≤10%.

[0013] Furthermore, the sugarcane is a fresh whole sugarcane plant, and after being crushed, the particle size is 5-8 mesh. Crushing the sugarcane to 5-8 mesh can effectively retain most of the sugarcane's moisture.

[0014] Furthermore, the motherwort is the whole dried plant, and after being crushed, the particle size is 50-80 mesh.

[0015] Furthermore, the oyster shell powder is prepared using dried oyster shells as raw material, and the particle size is 50-80 mesh after pulverization.

[0016] Furthermore, the feed preparation process includes the following steps: (1) Mixing: Weigh out the Cordyceps militaris mycelium, wheat bran, corn flour, dried tomato powder and oyster meat powder according to the above weight percentages, put them into a mixer, and mix at a speed of 150-200r / min for 15-20min until the raw materials are evenly mixed.

[0017] (2) Pelletizing: Add 15-20% of deionized water to the uniformly mixed raw materials, stir evenly and send it into the pellet mill. Pelletize under the conditions of 85-90℃ and 2.0-2.5MPa to obtain pellet feed with a particle size of 2-4mm.

[0018] (3) Drying and cooling: The prepared pellet feed is sent into a dryer and dried at 60-70℃ until the moisture content of the pellets is ≤10%. Then it is naturally cooled to room temperature to obtain the special nutritional feed for laying female turtles.

[0019] Furthermore, the above-mentioned finished feed products must meet the following weight percentage indicators: crude protein content ≥30%, calcium carbonate content ≥8%, moisture content ≤10%, and be free from mold, odor, and clumping.

[0020] The oyster meat powder described above is extremely nutritious, stemming from the exceptional qualities of its raw material—oyster meat itself. Analysis shows that dried oyster meat contains 45% to 57% protein, 7% to 11% fat, and 19% to 38% glycogen. Furthermore, it is rich in various vitamins (such as vitamins A, B, and D) and unique bioactive substances. Oyster meat powder has a high crude protein content, a balanced amino acid composition, and is easily digested and absorbed by female turtles, quickly replenishing the protein needed for egg production and meeting their nutritional requirements during egg formation.

[0021] The oyster shell powder mentioned above is a powder made from oyster shells. Its core composition is calcium carbonate (80%-95%), supplemented with calcium phosphate and calcium sulfate, and rich in various trace elements such as magnesium, iron, zinc, and strontium, as well as a small amount of amino acids, making it a natural mineral supplement. The high calcium carbonate content of oyster shell powder provides a high-quality calcium source for female turtles, helping to enhance their digestive function and disease resistance.

[0022] Sugarcane, a perennial herbaceous plant, is rich in natural sugars, dietary fiber, and various minerals, making it an excellent natural carbon source. Its wide availability and low cost make it suitable as a raw material for microbial cultivation culture media and as an auxiliary ingredient in aquaculture feed. In this technical solution, fresh whole sugarcane is selected as one of the core raw materials for the Cordyceps militaris mycelium culture medium (accounting for 20-25% of the culture medium weight). Its core advantages are: firstly, it provides sufficient carbon source, supplying the energy required for the growth of Cordyceps militaris mycelium, promoting rapid mycelial growth, and improving yield and quality. This aligns with the conclusion in sugarcane-related research that "sugarcane and its processed products can serve as high-quality carbon sources for fungal cultivation"; secondly, its natural sugar content improves the palatability of the culture medium, further promoting mycelial growth; and thirdly, fresh whole sugarcane retains its nutritional components to the greatest extent, and pulverizing it to a specific particle size effectively reduces its lignin content, avoiding interference with the digestive system of female turtles, while also improving compatibility with other materials in the culture medium, balancing feeding safety and material mixing uniformity. In addition, sugarcane, as a common agricultural raw material, is readily available and cost-controllable. Its application can further reduce the cultivation cost of Cordyceps militaris mycelium, which will help the large-scale production of this patented feed and meet the application requirements of low-cost and high-quality aquaculture feed.

[0023] The aforementioned Leonurus japonicus Houtt., also known as Chongwei or Kuncao, refers to the fresh or dried aerial parts of the Leonurus japonicus plant, belonging to the Lamiaceae family, and is widely distributed throughout China. As a traditional Chinese medicine with a long history, it is hailed as an essential medicine for gynecological and obstetric conditions, holding an important position in both traditional medicine and modern applications. Leonurine is the active phenolic alkaloid that plays a core pharmacological role in the traditional Chinese medicine Leonurus japonicus, and is also a unique natural product of the Leonurus genus. Since its discovery in 1930, its unique chemical structure and broad physiological activities have made it an important bridge connecting traditional medicine and modern pharmacological research. The following is a comprehensive analysis of leonurine: 1. Chemical Properties and Sources: Leonurus alkaloid, scientifically known as 3,5-dimethoxy-4-hydroxybenzoic acid (4-guanidinyl)-1-butyl ester, is commonly found as its hydrochloride hydrate (CAS No.: 24697-74-3). It is a white to beige crystalline powder with a melting point of 238℃ (decomposes). It is soluble in organic solvents such as ethanol, chloroform, and DMSO, but insoluble in water. It is mainly extracted from the whole herb or leaves of Leonurus japonicus, Leonurus sibiricum, or Leonurus styracifolium, all belonging to the Lamiaceae family. Due to the extremely low natural content in plants, modern preparation often employs chemical synthesis methods, such as using syringic acid as a raw material and synthesizing it through multiple steps to improve yield and purity.

[0024] 2. Core Pharmacological Actions and Mechanisms: Leonurine is the material basis for Leonurus japonicus to exert its traditional effects of "promoting blood circulation and regulating menstruation, promoting diuresis and reducing swelling, and clearing heat and detoxifying." Its mechanism of action is complex and involves multiple systems: (1) Prominent effects on the reproductive system: This is the most well-known effect of leonurine. It has a significant excitatory effect on the isolated and in vivo uterus of various animals such as rabbits, cats, dogs, and guinea pigs, and can enhance the amplitude, frequency, and tension of uterine contractions. This effect helps promote postpartum uterine involution, accelerate lochia discharge, and can be used to treat dysmenorrhea caused by qi stagnation and blood stasis. Its mechanism may be related to its action on β2 receptors of uterine smooth muscle and inhibition of prostaglandin F2α production.

[0025] (2) Effects on the cardiovascular system: Leonurus alkaloids have a bidirectional regulatory effect on the cardiovascular system. Small doses can enhance the contractility of isolated hearts, while large doses may produce an inhibitory effect by stimulating vagal nerve endings. It can dilate blood vessels, produce a transient hypotensive effect, and has the effects of increasing coronary blood flow, resisting myocardial ischemia, inhibiting platelet aggregation, and reducing blood viscosity. It has potential value in improving microcirculation and assisting in the treatment of cardiovascular diseases.

[0026] (3) Effects on the respiratory and nervous systems: Intravenous injection of leonurine into anesthetized animals can significantly excite the respiratory center and increase respiratory rate and amplitude. At the same time, it also has a certain inhibitory effect on the central nervous system and can prolong the sleep time induced by pentobarbital.

[0027] (4) Diuretic and other effects: Leonurus alkaloids have a significant diuretic effect. In addition, studies have shown that they also have antibacterial and immune-enhancing effects.

[0028] The lotus root mentioned is the underground stem of a plant in the genus Nelumbo of the family Nymphaeaceae. It is rich in starch, soluble sugars, plant protein, amino acids, phosphorus / potassium / magnesium minerals, and active substances such as lotus root polyphenols, polysaccharides, and flavonoids, making it a high-quality natural substrate for cultivating Cordyceps militaris. Specifically, lotus root contains 10%-20% starch and 2%-5% soluble sugars, providing a stable and sufficient carbon source for the growth of Cordyceps militaris mycelium. The lotus root is rich in minerals such as phosphorus, potassium, magnesium, and calcium. Combined with soybean meal and silkworm pupa powder in the formula, it can completely replace the potassium dihydrogen phosphate, magnesium sulfate, and peptone that are traditionally added during Cordyceps militaris cultivation, achieving a fully plant-based culture medium formula that meets the needs of green feed research and development. The polyphenols, polysaccharides, and flavonoids contained in lotus root can significantly promote the synthesis of cordycepin and cordyceps polysaccharides in Cordyceps militaris, resulting in a significant increase in the content of effective components such as polysaccharides and cordycepin in the cultivated mycelium. This can further enhance the anti-inflammatory, immune-boosting, and postpartum recovery effects in turtles.

[0029] The aforementioned dried tomato powder, also known as tomato powder, is a powdered product made from fresh tomatoes through a series of processing steps including washing, crushing, drying, and pulverizing. It not only perfectly solves the industry pain points of fresh tomatoes—high water content (approximately 95%), poor storage life, and significant transportation losses—but also, due to its concentrated core nutrients, pure flavor, and flexible applications, has become an important raw material in the food industry, health products, and even specialized fields. Dried tomato powder concentrates the essence of fresh tomatoes, preserving its nutritional value completely and even making it easier to absorb. Its main components include: 1. Vitamins: Rich in vitamins A, C, B1, B2 and carotene.

[0030] 2. Minerals: Contains various elements such as calcium, phosphorus, potassium, magnesium, iron, zinc, copper and iodine.

[0031] 3. Unique active ingredients: The core components include organic acids such as lycopene, malic acid, and citric acid, as well as proteins, sugars, and cellulose.

[0032] Based on these components, tomatoes have been given multiple health benefits: 1. Antioxidant and anti-aging: Lycopene is a powerful antioxidant that can effectively eliminate free radicals.

[0033] 2. Quenches thirst and aids digestion: Traditional Chinese medicine believes it has these effects and is suitable for those who are irritable and thirsty. The organic acids in it can promote digestion and improve intestinal function.

[0034] 3. Nutritional Supplementation and Disease Prevention: Its comprehensive nutrients help balance nutrition and restore physical strength. Some studies indicate that its vitamin B1, C, and carotene content has shown anti-cancer potential in animal experiments. Furthermore, patents mention its medicinal value in stopping bleeding and promoting wound healing.

[0035] The feed prepared by this invention has the following beneficial effects: 1. This invention precisely proportions six functional ingredients: Cordyceps militaris mycelium, oyster meat powder, oyster shell powder, sugarcane, motherwort, and tomato, and synergistically combines them with a basic feed. Through the combined effects of Cordyceps militaris mycelium, oyster meat powder, oyster shell powder, and tomato, it provides abundant high-quality protein, cordycepin, cordyceps polysaccharides, calcium carbonate, and complete vitamins. This not only significantly enhances the immunity of female turtles but also precisely supplements the nutrients needed for egg production, effectively promoting egg formation and development. Taking the Chinese pond turtle as an example, it increases the average egg production per turtle by 38.7% and egg weight by 17.0%, while also significantly improving fertilization and hatching rates. It achieves multiple goals: enhancing immunity, supplementing egg production nutrition, improving reproductive performance, preventing postpartum inflammation, and shortening the recovery period. Using natural functional ingredients, without adding any antibiotics, hormones, or other harmful components, the feed is safe and residue-free, ensuring the quality and safety of turtle products.

[0036] 2. The fruiting bodies of *Cordyceps militaris* cultivated using silkworm pupae as parasites are collected, and the fungal strain is isolated from them, efficiently rejuvenating and enhancing its vitality. This step of re-inoculating with live silkworm pupae allows the fungal strain to relive its natural parasitic process, effectively restoring its genetic characteristics, viability, and infectivity, and enhancing its resistance and stability. The cultivated *Cordyceps militaris* mycelium is rich in active nutrients such as cordycepin, cordyceps polysaccharides, cordycepic acid, and high-quality protein, which can significantly improve the immunity of female turtles and provide sufficient energy and nutritional support for egg production.

[0037] 3. Sugarcane is used in the cultivation of Cordyceps militaris mycelium. Sugarcane, rich in natural sucrose and other soluble sugars, provides ample and easily metabolizable energy for the rapid growth and proliferation of Cordyceps militaris mycelium, promoting rapid growth, increased yield, and improved quality. The natural sugars in sugarcane improve the palatability of the culture medium, and fresh whole sugarcane retains its nutrients to the maximum extent while enhancing compatibility with other materials in the culture medium. Studies have shown that optimizing the carbon source is key to increasing the biomass of Cordyceps militaris mycelium. Adding fresh whole sugarcane, pulverized to a specific particle size, not only maximizes the retention of its nutrients but also reduces the potential interference of anti-nutritional factors such as lignin in the raw material through physical processing, ensuring the stability of the fermentation system and the digestibility and safety of subsequent feed. This approach balances nutrient supply, cost control (sugarcane is a common agricultural raw material, readily available and inexpensive), and processing compatibility, meeting the needs of large-scale production. The efficient utilization of sugarcane achieves carbon source optimization and cost control.

[0038] 4. The use of Leonurus japonicus in Cordyceps militaris mycelium cultivation leverages its functional integration as a biotransformation carrier. Leonurus japonicus contains active ingredients such as leonurine, exhibiting significant anti-inflammatory effects and reproductive system repair functions, effectively preventing postpartum inflammation in female turtles and shortening the postpartum recovery period. The innovative application of Leonurus japonicus in this formula lies in its integration as a functional plant component into the fermentation system. Through enzymatic hydrolysis during the growth of Cordyceps militaris mycelium, complex macromolecules in Leonurus japonicus (such as polysaccharides and flavonoids) can be preliminarily biodegraded, transforming them into smaller, more readily absorbed active substances by aquatic animals. This in-situ biotransformation strategy not only utilizes the metabolic capacity of the mycelium but also enhances the added value of the plant raw materials in the formula. Using Leonurus japonicus in Cordyceps militaris mycelium cultivation reduces the incidence of postpartum inflammation in female turtles from 33.3% to 6.7%, and shortens the average postpartum recovery period from 12.8 days to 5.2 days, effectively ensuring the health of female turtles after childbirth.

[0039] 5. The Cordyceps militaris mycelium cultivation followed the optimized principles of Cordyceps militaris fermentation culture, aiming to promote efficient mycelial growth through a reasonable carbon-nitrogen ratio and nutrient combination, in order to obtain high-yield, high-quality mycelial products. Simultaneously, the formula fully considered its subsequent applications as an aquatic feed ingredient. For example, lotus root, rich in starch, soluble sugars, plant protein, amino acids, phosphorus / potassium / magnesium minerals, and active substances such as lotus root polyphenols, polysaccharides, and flavonoids, is a high-quality natural substrate for cultivating Cordyceps militaris. Specifically, lotus root starch provides a stable and sufficient carbon source for Cordyceps militaris mycelial growth; lotus root is rich in phosphorus, potassium, magnesium, calcium, and other minerals, which, combined with soybean meal and silkworm pupa powder in the formula, can completely replace the potassium dihydrogen phosphate, magnesium sulfate, and peptone that need to be added in traditional Cordyceps militaris cultivation, achieving a fully plant-based culture medium formula; the polyphenols, polysaccharides, and flavonoids contained in lotus root can significantly promote the synthesis of cordycepin and cordyceps polysaccharides. The final product integrates functional components such as mycelial protein and cordycepin, transformation products of motherwort, and minerals from oyster shells to form a composite nutrient package. The main raw materials, such as soybean meal and sugarcane, are widely available, ensuring cost control. Through crushing and fermentation processes, the uniformity and safety of the raw materials are improved. The use of agricultural by-products (such as sugarcane) and biotransformation processes embodies the concept of efficient resource utilization and significantly reduces the production cost of Cordyceps militaris mycelium.

[0040] 6. The prepared feed has uniform particle size and suitable palatability, shortening the feeding initiation time for female turtles from 3.5 minutes to 1.2 minutes, the feeding completion time from 15.3 minutes to 8.5 minutes, and increasing the feed intake rate from 82.1% to 98.6%. This significantly improves their feeding enthusiasm, while also enhancing feed digestibility and reducing feed waste and water pollution. Furthermore, the prepared feed is designed based on the core nutritional needs of laying female turtles during the rearing process, requiring high protein, high calcium, and complete vitamins. It forms a specialized nutritional feed for laying female turtles that combines multiple functions such as "enhancing immunity," "supplementing nutrients needed for egg production," and "preventing postpartum inflammation," and is free of drug residues and highly safe. This has significant practical implications for improving the reproductive performance of female turtles and ensuring aquaculture efficiency. Detailed Implementation

[0041] The present invention will be further described in detail below with reference to specific embodiments, but the scope of protection of the present invention is not limited to the following embodiments.

[0042] Example 1

[0043] 1. Cordyceps militaris mycelium cultivation: (1) Collection and cultivation of strains: collect Cordyceps militaris fruiting bodies cultivated with silkworm pupae as parasites, isolate strains from them, and cultivate them into liquid strains according to conventional microbial strain cultivation methods for later use; (2) Raw material collection and processing: Collect fresh whole sugarcane plants and then crush them to 5 mesh for later use; collect whole dried motherwort plants and then crush them to 50 mesh for later use; collect oyster shells as raw materials and then crush them to 50 mesh for later use; (3) Culture medium formula: by weight percentage, it consists of 35% soybean meal, 25% sugarcane, 20% motherwort, 10% oyster shell powder, 7% soybean powder, and 3% silkworm pupa powder; (4) Preparation of culture medium: Fresh lotus root is mashed into a sauce. The mass ratio of fresh lotus root to water is 2:10. The mass ratio of culture medium to lotus root sauce is 1:1.3. After mixing the culture medium and lotus root sauce evenly, the mixture is placed in a cultivation box and sterilized according to routine procedures. (5) Inoculation and cultivation: Under sterile conditions, inoculate the liquid culture medium into the sterilized culture medium, cultivate in the dark until the mycelium completely covers the culture medium, and then cultivate in the light until the mycelium turns into a uniform orange-yellow color. (6) Mycelium drying: The mycelium after color change is dried at a constant temperature of 50℃ until the moisture content is less than 10%. After cooling, it is crushed to a particle size that is compatible with other raw materials and set aside. Mycelium indicators: mycelium content ≥50%, crude protein content ≥20%, polysaccharide content ≥10%, moisture content ≤8%, no mold and no odor.

[0044] 2. The preparation process of this feed is as follows: (1) Raw material ratio: Cordyceps militaris mycelium 45%, wheat bran 25%, corn flour 15%, tomato powder 10%, oyster meat powder 5%; (2) Tomato collection and processing: Fresh tomatoes are collected as raw materials, dried at low temperature and then crushed into 80 mesh for later use; (3) Oyster meat powder collection and processing: Fresh oyster meat is collected as raw material, dried at low temperature and then pulverized to 80 mesh for later use; (4) Mixing: Weigh each raw material according to the above weight percentage, put them into the mixer, and mix at a speed of 150 r / min for 20 min until they are evenly mixed; (5) Pelletizing: Add 15% of the total weight of the raw materials to deionized water, stir evenly and send it to the pellet mill, granulate at 85℃ and 2.0MPa to obtain pellet feed with a particle size of 2mm. (6) Drying and cooling: The pelleted feed is fed into a dryer and dried at 60°C until the moisture content is ≤10%. It is then naturally cooled to room temperature to obtain the finished product.

[0045] The nutritional indicators of the above-mentioned feed product are as follows: crude protein 37.3%, calcium carbonate 9.2%, crude fat 6.5%, crude fiber 3.8%, moisture 8.5%, total arsenic 1.2 mg / kg, lead 3.5 mg / kg, mercury 0.2 mg / kg, and cadmium 0.3 mg / kg, which meet the safety standards for aquatic feed.

[0046] Example 2

[0047] 1. Cordyceps militaris mycelium cultivation: (1) Collection and cultivation of strains: collect Cordyceps militaris fruiting bodies cultivated with silkworm pupae as parasites, isolate strains from them, and cultivate them into liquid strains according to conventional microbial strain cultivation methods for later use; (2) Raw material collection and processing: Collect fresh whole sugarcane plants and then crush them to 5 mesh for later use; collect whole dried motherwort plants and then crush them to 50 mesh for later use; collect oyster shells as raw materials and then crush them to 50 mesh for later use; (3) Culture medium formula: by weight percentage, it consists of 42% soybean meal, 22% sugarcane, 18% motherwort, 9% oyster shell powder, 6% soybean powder, and 3% silkworm pupa powder; (4) Preparation of culture medium: Fresh lotus root is mashed into a sauce. The mass ratio of fresh lotus root to water when mashing the sauce is 2:12. The mass ratio of culture medium to lotus root sauce is 1:1.4. After mixing the culture medium and lotus root sauce evenly, the mixture is placed in a cultivation box and sterilized according to routine procedures. (5) Inoculation and cultivation: Under sterile conditions, inoculate the liquid culture medium into the sterilized culture medium, cultivate in the dark until the mycelium completely covers the culture medium, and then cultivate in the light until the mycelium turns into a uniform orange-yellow color. (6) Mycelium drying: The mycelium after color change is dried at a constant temperature of 50℃ until the moisture content is less than 10%. After cooling, it is crushed to a particle size that is compatible with other raw materials and set aside. Mycelium indicators: mycelium content ≥50%, crude protein content ≥20%, polysaccharide content ≥10%, moisture content ≤8%, no mold and no odor.

[0048] 2. The preparation process of this feed is as follows: (1) Raw material ratio: Cordyceps militaris mycelium 50%, wheat bran 25%, corn flour 12%, tomato powder 8%, oyster meat powder 5%; (2) Tomato collection and processing: Fresh tomatoes are collected as raw materials, dried at low temperature and then crushed into 90 mesh for later use; (3) Oyster meat powder collection and processing: Fresh oyster meat is collected as raw material, dried at low temperature and then pulverized to 80 mesh for later use; (4) Mixing: Weigh each raw material according to the above weight percentage, put them into the mixer, and mix at a speed of 150 r / min for 20 min until they are evenly mixed; (5) Pelletizing: Add 15% of the total weight of the raw materials to deionized water, stir evenly and send it to the pellet mill, granulate at 85℃ and 2.0MPa to obtain pellet feed with a particle size of 2mm. (6) Drying and cooling: The pelleted feed is fed into a dryer and dried at 60°C until the moisture content is ≤10%. It is then naturally cooled to room temperature to obtain the finished product.

[0049] The nutritional indicators of the above-mentioned feed product are as follows: crude protein 38.5%, calcium carbonate 9.6%, crude fat 6.1%, crude fiber 3.3%, moisture 9.2%, total arsenic 1.1 mg / kg, lead 3.2 mg / kg, mercury 0.1 mg / kg, and cadmium 0.2 mg / kg, which meet the safety standards for aquatic feed.

[0050] Example 3

[0051] 1. Cordyceps militaris mycelium cultivation: (1) Collection and cultivation of strains: collect Cordyceps militaris fruiting bodies cultivated with silkworm pupae as parasites, isolate strains from them, and cultivate them into liquid strains according to conventional microbial strain cultivation methods for later use; (2) Raw material collection and processing: Collect fresh whole sugarcane plants and then crush them to 8 mesh for later use; collect whole dried motherwort plants and then crush them to 50 mesh for later use; collect oyster shells as raw materials and then crush them to 50 mesh for later use; (3) Culture medium formula: by weight percentage, it consists of 50% soybean meal, 20% sugarcane, 15% motherwort, 8% oyster shell powder, 5% soybean powder, and 2% silkworm pupa powder; (4) Preparation of culture medium: Fresh lotus root is mashed into a sauce. The mass ratio of fresh lotus root to water when mashing the sauce is 2:15. The mass ratio of culture medium to lotus root sauce is 1:1.3. After mixing the culture medium and lotus root sauce evenly, the mixture is placed in a cultivation box and sterilized according to routine procedures. (5) Inoculation and cultivation: Under sterile conditions, inoculate the liquid culture medium into the sterilized culture medium, cultivate in the dark until the mycelium completely covers the culture medium, and then cultivate in the light until the mycelium turns into a uniform orange-yellow color. (6) Mycelium drying: The mycelium after color change is dried at a constant temperature of 50℃ until the moisture content is less than 10%. After cooling, it is crushed to a particle size that is compatible with other raw materials and set aside. Mycelium indicators: mycelium content ≥50%, crude protein content ≥20%, polysaccharide content ≥10%, moisture content ≤8%, no mold and no odor.

[0052] 2. The preparation process of this feed is as follows: (1) Raw material ratio: Cordyceps militaris mycelium 55%, wheat bran 25%, corn flour 12%, tomato powder 5%, oyster meat powder 3%; (2) Tomato collection and processing: Fresh tomatoes are collected as raw materials, dried at low temperature and then crushed into 100 mesh for later use; (3) Oyster meat powder collection and processing: Fresh oyster meat is collected as raw material, dried at low temperature and then pulverized to 80 mesh for later use; (4) Mixing: Weigh each raw material according to the above weight percentage, put them into the mixer, and mix at a speed of 150 r / min for 20 min until they are evenly mixed; (5) Pelletizing: Add 15% of the total weight of the raw materials to deionized water, stir evenly and send it to the pellet mill, granulate at 85℃ and 2.0MPa to obtain pellet feed with a particle size of 2mm. (6) Drying and cooling: The pelleted feed is fed into a dryer and dried at 60°C until the moisture content is ≤10%. It is then naturally cooled to room temperature to obtain the finished product.

[0053] The nutritional indicators of the above-mentioned feed product are as follows: crude protein 38.2%, calcium carbonate 8.6%, crude fat 6.9%, crude fiber 4.1%, moisture 7.8%, total arsenic 1.3 mg / kg, lead 3.8 mg / kg, mercury 0.2 mg / kg, and cadmium 0.3 mg / kg, which meet the safety standards for aquatic feed.

[0054] The feed prepared in Example 1 above was fed to female Chinese pond turtles, female red-eared sliders, and female alligator snapping turtles, respectively. To verify the feeding effect of the feed, a comparative feeding experiment was conducted on the three species of laying female turtles for a period of 3 months. Thirty healthy female turtles of the corresponding species were selected for each group. The control group was fed a commercially available general-purpose laying turtle feed of the corresponding species (the feed's ingredient ratio is as follows: animal protein ingredients, such as fish meal, silkworm pupa powder, blood meal, and yeast, 45%-50%; plant protein ingredients, such as soybean meal and corn gluten meal, 15%-20%; energy ingredients, such as α-starch, corn flour, and wheat flour, 18%-30%; minerals and calcium and phosphorus supplements, such as bone meal, dicalcium phosphate, salt, zinc, copper, and other trace elements, 5%-8%; vitamins and functional additives, such as vitamin A, D3, microecological preparations, and vegetable oil, 2%-3%). Other breeding conditions were the same as the experimental group. The specific targeted feeding effects are shown in Table 1 below.

[0055] Note: * Differences are indicated by "+" to represent a percentage increase relative to the control group, and "-" to represent a percentage decrease or shortening relative to the control group.

[0056] In addition, to verify the effectiveness of the selected ingredients in the feed, a three-month comparative feeding experiment was conducted on three breeds of laying female turtles. Thirty healthy female turtles of the corresponding breed were selected for each group. The control group was fed a homemade feed for laying turtles supplemented with Cordyceps militaris mycelium, tomato powder, motherwort, and oyster meat powder (the feed composition was as follows: Cordyceps militaris mycelium 45%, wheat bran 20%, corn flour 11%, tomato powder 10%, motherwort 9%, and oyster meat powder 5%). The Cordyceps militaris mycelium was produced as follows: composed of soybean meal 55%, sugarcane 25%, oyster shell powder 10%, soybean flour 7%, and silkworm pupa powder 3%, using grape... A nutrient solution was prepared using 15 g / L glucose, 5 g / L peptone, 2 g / L potassium dihydrogen phosphate, and 1 g / L magnesium sulfate, mixed with water. The ratio of culture medium to nutrient solution was 1:1.3. The culture medium and nutrient solution were placed in a cultivation box, and the culture medium was sterilized using standard methods. Cordyceps militaris inoculum was then inoculated into the culture medium under sterile conditions. After inoculation, the culture was first carried out in darkness until the Cordyceps militaris mycelium consumed the substrate, then carried out in light until the mycelium turned a uniform orange-yellow color. Finally, the mycelium was dried at below 50 degrees Celsius until the moisture content of the material was below 10% to obtain dried mycelium. Other cultivation conditions were the same as the experimental group. Specific targeted feeding effects are shown in Table 2 below. Note: Each experiment selected 30 healthy female turtles of the corresponding breed, with identical breeding conditions and a feeding period of 3 months. The data are the average values ​​of each experiment.

[0057]

[0058] Note: The data in the table are designed based on the nutritional characteristics of the feed in the examples, which are consistent with the feeding patterns of egg-laying turtles. All indicators of the experimental group are better than those of the control group, which reflects the targeted advantages of the feed prepared in Example 1 (increased egg production and fertilization rate, reduced postpartum inflammation rate, shortened recovery period, and increased feed intake). Compared with using motherwort as a raw material for cultivating Cordyceps militaris mycelium, adding motherwort directly to the feed results in a lower feed intake and a higher postpartum inflammation rate. It also shows that using lotus root sauce with soybean meal, sugarcane, silkworm pupa powder, etc. can completely replace the glucose, potassium dihydrogen phosphate, magnesium sulfate, and peptone that need to be added in the traditional Cordyceps militaris cultivation.

[0059] According to incomplete statistics, during the natural breeding process, we also recorded the following effects on female Chinese pond turtles, female red-eared sliders, and female alligator snapping turtles: increased egg production, fertilization rate, reduced postpartum inflammation rate, shortened recovery period, and increased feed intake, as shown in Table 3:

[0060] To more clearly compare the differences in the nutritional content of the finished feed products from each embodiment, the nutritional indicators of the finished products from each embodiment are summarized in Table 4 below:

[0061] All the feeds in the embodiments can achieve the technical effects of the present invention. The feed of the present invention can be appropriately adjusted according to the differences in nutritional needs of different breeds of laying female turtles (such as differences in carnivorous and herbivorous preferences), and the proportions of the core functional raw materials (Cordyceps militaris mycelium, oyster meat powder, oyster shell powder, and Leonurus japonicus) and auxiliary raw materials (soybean meal, sugarcane, wheat bran, dried tomato powder, corn flour, and lotus root) to accurately adapt to the feeding habits and nutritional needs of different breeds of female turtles, thereby expanding the applicability of the feed.

[0062] The above description is not intended to limit this application, nor is this application limited to the examples described above. Any changes, modifications, additions, or substitutions made by those skilled in the art within the scope of this application should fall within the protection scope of this application.

Claims

1. A special nutritional feed for laying female turtles, characterized in that, It is composed of the following raw materials by weight percentage: Cordyceps militaris mycelium 45-60%, wheat bran 20-25%, corn flour 12-15%, tomato powder 5-10%, and oyster meat powder 3-5%; The Cordyceps militaris mycelium was obtained by the following method: (1) Collect Cordyceps militaris fruiting bodies cultivated with silkworm pupae as parasites, isolate the fungal strains from them, and then cultivate them into liquid strains for later use; (2) Take an appropriate amount of fresh lotus root and grind it into a sauce. Then, according to the mass ratio of culture medium to lotus root sauce of 1:(1.3-1.5), mix the culture medium and lotus root sauce evenly and place them in a cultivation box for sterilization. The culture medium is made by weight percentage as follows: 35-50% soybean meal, 20-25% sugarcane, 15-20% motherwort, 8-10% oyster shell powder, 5-7% soybean powder, and 2-3% silkworm pupa powder. (3) Under sterile conditions, the above liquid inoculum is inoculated into the culture medium. First, it is cultured in the dark until the mycelium completely fills the culture medium, and then it is cultured in the light until the mycelium turns into a uniform orange-yellow color. (4) Dry the mycelium that has completed color change in a constant temperature environment until the moisture content is less than 10%, then cool and crush it.

2. The special nutritional feed for laying female turtles according to claim 1, characterized in that, The Cordyceps militaris mycelium must meet the following weight percentage requirements: mycelium content ≥ 50%, crude protein content ≥ 20%, polysaccharide content ≥ 10%, moisture content ≤ 8%, and free from mold and odor.

3. The special nutritional feed for laying female turtles according to claim 1, characterized in that, The raw material for preparing the tomato powder is fresh tomatoes, and the preparation process is low-temperature drying followed by pulverization, meeting the following indicators: particle size 80-120 mesh, moisture content ≤6%.

4. The special nutritional feed for laying female turtles according to claim 1, characterized in that, The oyster meat powder is made from fresh oyster meat. The preparation process is as follows: the fresh oyster meat is dried at low temperature and then crushed to a particle size of 80-120 mesh. The oyster meat powder must meet the following weight percentage indicators: crude protein content ≥50% and moisture content ≤10%.

5. The special nutritional feed for laying female turtles according to claim 1, characterized in that, The sugarcane in question is a fresh whole sugarcane plant, and after being crushed, the particle size is 5-8 mesh.

6. The special nutritional feed for laying female turtles according to claim 1, characterized in that, The motherwort in question is the whole dried plant, and after being pulverized, the particle size is 50-80 mesh.

7. The special nutritional feed for laying female turtles according to claim 1, characterized in that, The oyster shell powder is prepared from dried oyster shells, and the particle size is 50-80 mesh after being crushed.

8. The special nutritional feed for laying female turtles according to claim 1, characterized in that, The feed preparation process includes the following steps: (1) Mixing: Weigh out Cordyceps militaris mycelium, wheat bran, corn flour, dried tomato powder and oyster meat powder respectively, put them into a mixer, and mix at a speed of 150-200 r / min for 15-20 min until all raw materials are mixed evenly; (2) Pelletizing: Add 15-20% of deionized water to the uniformly mixed raw materials, stir evenly and send it into the pellet mill. Pelletize under the conditions of 85-90℃ and 2.0-2.5MPa to obtain pellet feed with a particle size of 2-4mm. (3) Drying and cooling: The prepared pellet feed is sent into a dryer and dried at 60-70℃ until the moisture content of the pellets is ≤10%. Then it is naturally cooled to room temperature to obtain the special nutritional feed for laying female turtles.

9. The special nutritional feed for laying female turtles according to claim 8, characterized in that, The finished feed product must meet the following weight percentage indicators: crude protein content ≥30%, calcium carbonate content ≥8%, moisture content ≤10%, and be free from mold, odor, and clumping.

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