Preparation method of culture solution for promoting gonad development of pinctada maxima
By using a formula of extracts from traditional Chinese herbs such as Isatis root in the culture ponds of giant pearl oysters, the problem of poor gonadal development in giant pearl oysters has been solved, achieving the promotion of gonadal development and the improvement of survival rate, reducing the risk of disease, and generating economic benefits.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-24
- Publication Date
- 2026-03-24
AI Technical Summary
Environmental pollution affects the gonadal development of pearl oysters, leading to physiological dysfunction, decreased reproductive capacity, and impact on pearl formation quality and aquaculture efficiency. It also poses risks of disease and a decline in genetic diversity.
An extract formula consisting of Isatis root, honeysuckle, gentian, wolfberry, dodder seed, alisma, and anemarrhena is prepared by boiling, cooling, filtering, and high-pressure sterilization to create a culture medium that promotes the gonadal development of pearl oysters. This medium is then applied by sprinkling it into the aquaculture pond.
It promotes the development of gonads in pearl oysters, increases survival rate, inhibits harmful bacteria, reduces disease risk, improves pearl quality and farming efficiency, and is easy to operate and inexpensive.
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Figure CN121714643A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of giant pearl oyster aquaculture technology, and more specifically, to a method for preparing a culture medium that promotes gonadal development in giant pearl oysters. Background Technology
[0002] The large pearl oyster (Pinctada maxima jameson), also known as the white-lipped pearl oyster, belongs to the phylum Mollusca, class Lamellibranchia, order Pterioida, family Pteriidate, and genus Pinctada in animal taxonomy. It is currently the largest marine pearl oyster. Large pearl oysters are mainly distributed in the South and West Pacific Oceans. In my country, they are mainly found in the waters off Hainan Island, the Leizhou Peninsula, and the Xisha Islands; internationally, they are distributed in the coastal waters of Southeast Asian countries. Since the 1960s, my country has successfully conducted scientific research on key technologies for artificial seedling cultivation and nucleus insertion pearl cultivation of large pearl oysters, which has promoted the rapid development of marine pearl farming in my country. By the 1980s, my country had become a major marine pearl farming country in the world, and pearls cultivated from large pearl oysters have become the main cultured pearl species in my country, as well as one of the pillar industries for the development of the marine economy in Hainan, Guangdong, and Guangxi.
[0003] In recent years, with the rapid economic development of coastal areas and the intensive utilization of marine resources in my country, the marine ecological environment has been significantly impacted, and water pollution has become increasingly prominent. As a result, the large pearl oyster, an important species in marine pearl farming, suffers severe disruption to its physiological functions, particularly gonadal development, thus affecting its growth, reproduction, and pearl formation quality. Environmental pollution has led to a continuous increase in the levels of heavy metals, organic pollutants, and microplastics in seawater. These harmful substances accumulate in the oysters' bodies, disrupting endocrine balance, inhibiting gonadal development, reducing the oyster's reproductive capacity, and consequently weakening pearl quality. Simultaneously, eutrophication in aquaculture areas causes abnormal algal growth and insufficient dissolved oxygen, further deteriorating the oysters' living environment and leading to metabolic disorders and decreased immunity. Long-term exposure to polluted environments commonly results in poor gonadal development, delayed maturity, and decreased reproductive rates in large pearl oysters. This not only affects their normal pearl cultivation process but also severely restricts the sustainable development of my country's marine pearl industry.
[0004] Under increasingly severe environmental stress, the survival rate of large pearl oysters has also declined significantly. Recent survey data shows that the mortality rate of oysters in several major producing areas has been increasing year by year. Besides environmental degradation, the reasons for this phenomenon include excessively high farming density, improper water quality management, and frequent disease outbreaks. Increased pathogens and parasites in polluted waters easily lead to diseases such as hepatopancreatic necrosis and mantle lesions in shellfish, resulting in a significant increase in mortality and nucleus expulsion rates during pearl cultivation. Simultaneously, due to long-term artificial breeding and intensive selection, the genetic diversity of large pearl oyster populations has decreased, and germplasm degradation is prominent, manifested as decreased individual physical condition, weakened resistance, and slow growth. The combined effect of poor gonadal development and increased mortality has resulted in a sharp reduction in the number of healthy oysters suitable for pearl cultivation, directly affecting pearl yield and quality, and gradually diminishing the competitiveness of Chinese seawater pearls in the international market.
[0005] This invention can promote gonadal development, inhibit bacteria, scavenge free radicals, and maintain cell vitality. In addition, the abundant vitamins and minerals it contains can promote the growth and development of gonadal cells, and further promote the gonadal development of shellfish. Summary of the Invention
[0006] The present invention aims to solve the technical problems mentioned in the background art and provide a method for preparing a culture medium that promotes the gonadal development of pearl oysters.
[0007] To achieve the above objectives, the present invention provides the following technical solution: a method for preparing a culture medium to promote gonadal development in pearl oysters, comprising the following components: Isatis root extract, honeysuckle extract, gentian root extract, wolfberry extract, dodder seed extract, alisma extract, and anemarrhena extract; The amounts of each component added to the above culture medium are as follows: Isatis root extract 250-300 ml / L, honeysuckle extract 180-220 ml / L, gentian extract 140-200 ml / L, wolfberry extract 120-180 ml / L, dodder seed extract 100-150 ml / L, alisma extract 80-100 ml / L, anemarrhena extract 30-50 ml / L; After washing the above-mentioned raw materials for extraction, they were boiled in ultrapure water, cooled naturally after boiling, filtered through a 200-mesh sieve, and then sterilized under high temperature and high pressure to obtain the extracts. The above extract was sterilized under ultraviolet light and then added to the solution in a sterile operating room according to the reference dosage and mixed well to obtain a culture medium for promoting gonadal development of pearl oysters.
[0008] A further preferred method: After cleaning the Isatis root, rinse it 2-3 times with purified water, then put 500g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then sterilize the resulting solution at high temperature and high pressure to obtain the Isatis root extract.
[0009] A further optimized solution: After cleaning the honeysuckle, rinse it 2-3 times with purified water, then put 300g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then sterilize the resulting solution at high temperature and high pressure to obtain honeysuckle extract.
[0010] A further preferred method: After cleaning the gentian root, rinse it 2-3 times with purified water, then put 150g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then sterilize the resulting solution at high temperature and high pressure to obtain the gentian root extract.
[0011] A further preferred method: After cleaning the goji berries, rinse them 2-3 times with purified water. Then, put 100g into 1L of ultrapure water and boil for 30 minutes. After boiling, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then bring the volume of the resulting solution to 1L and sterilize it under high temperature and high pressure to obtain the goji berry extract.
[0012] A further optimized solution: After cleaning the dodder seeds, rinse them 2-3 times with purified water. Then, put 50g into 1L of ultrapure water and boil for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. After making up to 1L of the solution, sterilize it under high temperature and high pressure to obtain the dodder seed extract.
[0013] A further preferred method: After cleaning the Alisma plantago-aquatica, rinse it 2-3 times with purified water, then put 30g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then sterilize the resulting solution at high temperature and high pressure to obtain the Alisma plantago-aquatica extract.
[0014] A further preferred method: After cleaning Anemarrhena asphodeloides, rinse it 2-3 times with purified water, then put 50g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then sterilize the resulting solution at high temperature and high pressure to obtain Anemarrhena asphodeloides extract. Beneficial effects
[0015] 1. By adding traditional Chinese medicines such as Isatis indigotica and honeysuckle, along with other components, to the culture tank of giant pearl oysters, the optimal ratio of each component is used to promote the maturation of the gonads of giant pearl oysters. The added traditional Chinese medicine components not only stimulate the development of the gonads of giant pearl oysters, but also inhibit harmful bacteria in the external environment throughout the culture process, thereby improving their survival rate and achieving the dual benefits of promoting development and preventing diseases. 2. By selecting non-toxic and hormone-free Chinese herbal medicines, the use process is unlikely to cause environmental pollution or drug accumulation in the pearl oyster. Moreover, the operation process is simple and the raw material cost is low, making it highly valuable for practical promotion. 3. In summary, this method for preparing a culture medium that promotes gonadal development in giant pearl oysters further optimizes the formulation of the culture medium. It not only realizes the application of traditional Chinese medicine ingredients to promote gonadal development in giant pearl oysters, improving gonadal development efficiency and survival rate, and effectively avoiding the risk of disease outbreaks, but also provides solid theoretical support and practical reference for gonadal development promotion technology in large-scale giant pearl oyster farming. Attached Figure Description
[0016] Figure 1 This is a schematic diagram illustrating the effects of different culture media on the gonadal development and survival rate of pearl oysters. Detailed Implementation
[0017] The following will refer to the appendices in the embodiments of the present invention. Figure 1 The technical solutions in the embodiments of the present invention will be clearly and completely described.
[0018] Please see Figure 1 In this embodiment of the invention, a method for preparing a culture medium to promote gonadal development in pearl oysters includes the following components: Isatis root extract, honeysuckle extract, gentian root extract, wolfberry extract, dodder seed extract, alisma extract, and anemarrhena extract. The amounts of each component added to the above culture medium are as follows: Isatis root extract 250-300 ml / L, honeysuckle extract 180-220 ml / L, gentian extract 140-200 ml / L, wolfberry extract 120-180 ml / L, dodder seed extract 100-150 ml / L, alisma extract 80-100 ml / L, anemarrhena extract 30-50 ml / L; After washing the above-mentioned raw materials for extraction, they were boiled in ultrapure water, cooled naturally after boiling, filtered through a 200-mesh sieve, and then sterilized under high temperature and high pressure to obtain the extracts. The above extract was sterilized under ultraviolet light and then added and mixed in a sterile operating room according to the reference dosage to obtain a culture medium that promotes the gonadal development of pearl oysters. After cleaning the Isatis root, rinse it 2-3 times with purified water. Then, boil 500g in 1L of ultrapure water for 30 minutes, allow it to cool naturally, filter it through a 200-mesh sieve to remove residue, and then sterilize the resulting solution at high temperature and high pressure to obtain the Isatis root extract. After cleaning the honeysuckle, rinse it 2-3 times with purified water. Then, boil 300g in 1L of ultrapure water for 30 minutes, allow it to cool naturally, and then filter it through a 200-mesh sieve to remove residue. The resulting solution was brought to a final volume of 1L and then autoclaved to obtain honeysuckle extract. Gentian root was washed thoroughly and rinsed 2-3 times with purified water. 150g of the gentian root was then boiled in 1L of ultrapure water for 30 minutes, allowed to cool naturally, and filtered through a 200-mesh sieve to remove residue. The resulting solution was brought to a final volume of 1L and then autoclaved to obtain gentian root extract. Goji berries were washed thoroughly and rinsed 2-3 times with purified water. 100g of the goji berries was then boiled in 1L of ultrapure water for 30 minutes. After natural cooling, the solution was filtered through a 200-mesh sieve to remove residue. The resulting solution was then brought to a final volume of 1L and autoclaved to obtain the wolfberry extract. The dodder seeds were cleaned and rinsed 2-3 times with purified water. 50g of the dodder seeds were then boiled in 1L of ultrapure water for 30 minutes. After natural cooling, the solution was filtered through a 200-mesh sieve to remove residue. The resulting solution was then brought to a final volume of 1L and autoclaved to obtain the dodder seed extract. The alisma rhizome was cleaned and rinsed 2-3 times with purified water. Then, boil 30g in 1L of ultrapure water for 30 minutes, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then sterilize the resulting solution at a high temperature and high pressure to obtain the Alisma plantago-aquatica extract; after cleaning Anemarrhena asphodeloides, rinse it 2-3 times with purified water, then boil 50g in 1L of ultrapure water for 30 minutes, let it cool naturally, filter it through a 200-mesh sieve to remove the residue, and then sterilize the resulting solution at a high temperature and high pressure to obtain the Anemarrhena asphodeloides extract; The culture solution prepared in this invention is sprayed twice a week in the culture pond of giant pearl oysters at a dosage of 200-300 mL / m3. Under the premise of maintaining the stability of the culture water, it can promote the gonadal development of giant pearl oysters. In some embodiments, the above method is used to extract 250 ml / L of Isatis indigotica extract, 200 ml / L of Lonicera japonica extract, 150 ml / L of Gentiana scabra extract, 120 ml / L of Lycium barbarum extract, 150 ml / L of Cuscuta chinensis extract, 100 ml / L of Alisma plantago-aquatica extract, and 30 ml / L of Anemarrhena asphodeloides extract. After the above extracts are thoroughly mixed, they can be used in the pearl oyster culture pond. This embodiment is Example 1. In some embodiments, the above method is used to extract 280 ml / L of Isatis indigotica extract, 180 ml / L of Lonicera japonica extract, 140 ml / L of Gentiana scabra extract, 120 ml / L of Lycium barbarum extract, 150 ml / L of Cuscuta chinensis extract, 100 ml / L of Alisma plantago-aquatica extract, and 30 ml / L of Anemarrhena asphodeloides extract. After the above extracts are thoroughly mixed, they can be used in the pearl oyster culture pond. This embodiment is Example 2. In some embodiments, the above method is used to extract 260 ml / L of Isatis indigotica extract, 180 ml / L of Lonicera japonica extract, 140 ml / L of Gentiana scabra extract, 130 ml / L of Lycium barbarum extract, 150 ml / L of Cuscuta chinensis extract, 100 ml / L of Alisma plantago-aquatica extract, and 40 ml / L of Anemarrhena asphodeloides extract. After the above extracts are thoroughly mixed, they can be used in the pearl oyster culture pond. This embodiment is Example 3. In some embodiments, the above method is used to extract 280 ml / L of Isatis indigotica extract, 200 ml / L of Lonicera japonica extract, 140 ml / L of Gentiana scabra extract, 120 ml / L of Lycium barbarum extract, 150 ml / L of Cuscuta chinensis extract, 80 ml / L of Alisma plantago-aquatica extract, and 30 ml / L of Anemarrhena asphodeloides extract. After the above extracts are thoroughly mixed, they can be used in the pearl oyster culture pond. This embodiment is Example 4. Then, healthy and vigorous pearl oysters were selected and temporarily reared for 20 days. 1500 pearl oysters with an average weight of 146.3g were randomly divided into 5 groups, with 3 replicates per group and 100 oysters per replicate. Each group of pearl oysters was independently cultured in fiberglass tanks (cylindrical, 2m³ / tank), using a unified water source (water quality indicators: temperature 28-32℃, salinity 27‰-34‰, pH 7.5-8.5, dissolved oxygen ≥7.0 mg / L, ammonia nitrogen ≤0.2 mg / L, nitrite ≤0.02 mg / L). The culture solution prepared in the above example was then poured into the 5 pearl oyster rearing ponds. The culture medium application details for each group are as follows: Group A, serving as the control group, was treated with distilled water twice a week at a dose of 200 ml / L in the pearl oyster culture pond. Group B: The culture medium of Example 1 was sprayed twice a week into the pearl oyster culture pond at a dose of 200 ml / L. Group C: The culture medium of Example 2 was sprayed twice a week into the pearl oyster culture pond at a dose of 200 ml / L. Group D: The culture medium of Example 3 was sprayed twice a week into the pearl oyster culture pond at a dose of 200 ml / L. Group E: The culture medium of Example 4 was sprayed twice a week in the pearl oyster culture pond at a dose of 300 ml / L; The culture period was 15 weeks. During the culture, the gonadal development status and mortality rate of the pearl oysters were recorded. Experimental data are expressed as mean ± standard deviation. One-way ANOVA was performed on each indicator using SPSS 18.0 software, and multiple comparisons were also conducted. The experiment ended after 15 weeks of culture under different culture solutions. The gonadal development and survival rate results for each group are as follows: Figure 1 As shown, through Figure 1 It can be seen that the survival rate of experimental groups B to E, treated with the culture solution of this invention, all reached over 93%, significantly higher than that of the control group A, and the difference was statistically significant (P < 0.05). Among the experimental groups B to E, group E had the highest survival rate at 96.67%, followed by groups B, C, and D, but there was no significant difference in survival rate among the groups B to E. The number of individuals with mature gonads in experimental groups B to E, treated with the culture solution of this invention, all reached over 67, significantly higher than that of the control group A, and the difference was statistically significant (P < 0.01). Among the experimental groups B to E, group E had the highest number of individuals with mature gonads at 71, followed by groups B, C, and D, but there was no significant difference in survival rate among the groups B to E. Therefore, the culture solution designed in this invention can significantly improve the gonadal development of pearl oysters and increase their survival rate. Furthermore, the experiment further reveals that the rational combination of different components is beneficial for further improving the gonadal growth and development effect. In summary, by spraying non-toxic and side-effect-free Isatis indigotica extract, honeysuckle extract, and other formula components into the pearl oyster culture pond, the gonadal development of the pearl oyster can be further improved, and its survival rate can be increased. In addition, the operation of this invention is simple, reduces production costs, and has great economic potential.
[0019] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention. In addition, it should be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately. Furthermore, various different embodiments of the present invention can also be arbitrarily combined, as long as they do not violate the spirit of the present invention, they should also be regarded as the content disclosed by the present invention.
Claims
1. A method for preparing a culture medium to promote gonadal development in pearl oysters, characterized in that: It includes the following ingredients: Isatis root extract, honeysuckle extract, gentian root extract, wolfberry extract, dodder seed extract, alisma extract, and anemarrhena extract; The amounts of each component added to the above culture medium are as follows: Isatis root extract 250-300 ml / L, honeysuckle extract 180-220 ml / L, gentian extract 140-200 ml / L, wolfberry extract 120-180 ml / L, dodder seed extract 100-150 ml / L, alisma extract 80-100 ml / L, anemarrhena extract 30-50 ml / L; After washing the above-mentioned raw materials for extraction, they were boiled in ultrapure water, cooled naturally after boiling, filtered through a 200-mesh sieve, and then sterilized under high temperature and high pressure to obtain the extracts. The above extract was sterilized under ultraviolet light and then added to the solution in a sterile operating room according to the reference dosage and mixed well to obtain a culture medium for promoting gonadal development of pearl oysters.
2. The method for preparing a culture medium for promoting gonadal development in pearl oysters according to claim 1, characterized in that: After cleaning the Isatis root, rinse it 2-3 times with purified water. Then, put 500g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. Make up the volume of the obtained solution to 1L and then sterilize it under high temperature and high pressure to obtain the Isatis root extract.
3. The method for preparing a culture medium for promoting gonadal development in pearl oysters according to claim 1, characterized in that: After cleaning the honeysuckle, rinse it 2-3 times with purified water. Then, put 300g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. Make up the volume of the resulting solution to 1L and then sterilize it under high temperature and high pressure to obtain the honeysuckle extract.
4. The method for preparing a culture medium for promoting gonadal development in pearl oysters according to claim 1, characterized in that: After cleaning the gentian root, rinse it 2-3 times with purified water. Then, put 150g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. Make up the volume of the resulting solution to 1L and then sterilize it under high temperature and high pressure to obtain the gentian root extract.
5. The method for preparing a culture medium for promoting gonadal development in pearl oysters according to claim 1, characterized in that: After cleaning the goji berries, rinse them 2-3 times with purified water. Then, put 100g into 1L of ultrapure water and boil for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. Make up the volume of the resulting solution to 1L and then sterilize it under high temperature and high pressure to obtain the goji berry extract.
6. The method for preparing a culture medium for promoting gonadal development in pearl oysters according to claim 1, characterized in that: After cleaning the dodder seeds, rinse them 2-3 times with purified water. Then, put 50g into 1L of ultrapure water and boil for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. Make up the volume of the obtained solution to 1L and then sterilize it under high temperature and high pressure to obtain the dodder seed extract.
7. The method for preparing a culture medium for promoting gonadal development in pearl oysters according to claim 1, characterized in that: After cleaning the Alisma plantago-aquatica, rinse it 2-3 times with purified water. Then, put 30g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. Make up the volume of the obtained solution to 1L and then sterilize it under high temperature and high pressure to obtain the Alisma plantago-aquatica extract.
8. The method for preparing a culture medium for promoting gonadal development in pearl oysters according to claim 1, characterized in that: After cleaning Anemarrhena asphodeloides, rinse it 2-3 times with purified water. Then, put 50g into 1L of ultrapure water and boil it for 30 minutes. After boiling, let it cool naturally and filter it through a 200-mesh sieve to remove the residue. Make up the volume of the resulting solution to 1L and then sterilize it under high temperature and high pressure to obtain the Anemarrhena asphodeloides extract.