Methods for Synchronized Maturation of Sipunculus nudus and Their Applications

Abstract

This invention discloses a method for synchronizing the maturation of sperm and eggs of *Sipunculus nudus* and its application. This method ingeniously discloses a method for synchronizing the maturation of sperm and eggs of *Sipunculus nudus* by simulating the ebb and flow of tides and light in the daily breeding environment of *Sipunculus nudus*, resulting in better growth, survival rate, and egg production. Furthermore, this method provides physiological stimulation to mature *Sipunculus nudus* regarding sperm and egg release by repeatedly immersing them and gradually changing the salinity and temperature of seawater to promote egg and sperm production, while simultaneously ensuring a good hatching rate for the fertilized eggs. This method can be used in the artificial breeding of *Sipunculus nudus*, and it is not only reliable to implement but also has high reproductive efficiency and high survival rate.

Description

Technical Field

[0001] This invention relates to the field of aquaculture technology, and in particular to a method for synchronizing the maturation of sperm and eggs of Sipunculus nudus and its application. Background Technology

[0002] Delicious Sipunculid (a type of worm) Phascolosoma esculenta *Sipunculus nudus*, commonly known as "sea snail" or "mud snail," belongs to the phylum Sipunculidae, class Sipunculidae, and genus *Sipunculus*. Currently, *Sipunculus nudus* is widely distributed in provinces such as Zhejiang. Due to its high medicinal and edible value, a breeding industry for *Sipunculus nudus* has emerged in Fujian and Zhejiang, achieving good economic benefits. However, breeding *Sipunculus nudus* presents certain difficulties. Currently, most seedlings are obtained from natural resources, and artificially bred seedlings are rarely sold on the market. Collecting wild seedlings for breeding is not only costly but also carries the risk of decreased survival rates due to environmental changes when transferred to breeding areas. Furthermore, the number of eggs laid by wild *Sipunculus nudus* may fluctuate depending on their habitat, making it difficult to maintain optimal or high egg-laying efficiency. Therefore, developing a method for artificially breeding *Sipunculus nudus* is a highly significant and practically important research topic. Summary of the Invention

[0003] In view of this, the purpose of this invention is to propose a reliable, flexible, and efficient method for synchronizing the maturation of sperm and eggs of Sipunculus nudus and its application.

[0004] To achieve the above-mentioned technical objectives, the technical solution adopted by this invention is as follows:

[0005] A method for synchronizing the maturation of sperm and eggs of the palatable Sipunculus nudus, comprising:

[0006] S01. Select palatable Sipunculus worms that meet the preset requirements and classify them by sex.

[0007] S02. The Sipunculus nudus were divided into groups of 40, with a male-to-female ratio of 3:1, and then transferred into culture soil for breeding.

[0008] S03. Control the breeding environment of Sipunculus nudus according to the preset light duration and salinity to imitate the natural ecology. At the same time, the breeding environment of Sipunculus nudus is filled and drained twice a day to simulate the ebb and flow of tides.

[0009] S04. Stimulate the cultured Sipunculus nudus according to the preset method to induce them to release eggs or sperm, and then collect the sperm and eggs.

[0010] As one possible implementation, further, in this scheme S01, palatable Sipunculus nudus that meet the preset health requirements and weigh between 3 and 4 g are selected and classified according to sex.

[0011] As one possible implementation, further, in this scheme S02, the culture soil is prepared at a concentration of 15-65 g / m³. 2 Add fermented chicken manure as base fertilizer.

[0012] As one possible implementation method, further, in this scheme S02, the month in which the Sipunculus nudus is transferred into the culture soil for cultivation is October of each year;

[0013] In S04, when the cultured insects reach June of the following year, physiological stimulation is applied to the cultured Sipunculus nudus according to a preset method.

[0014] As a preferred implementation method, in S03 of this scheme, the water in the breeding environment of Sipunculus nudus is controlled at a salinity of 24±1 ppt and a pH of 6.5±0.2.

[0015] As a preferred implementation method, in S03 of this scheme, the light intensity of the breeding environment of Sipunculus nudus is controlled at a light duration of 9 to 12 hours per day.

[0016] As a preferred implementation method, in S03 of this scheme, diatoms are added to the water in the breeding environment of Sipunculus nudus, once every 15 days, at a rate of 10-50 g / m³ each time. 2 .

[0017] As a preferred implementation method, S03 of this scheme further includes: randomly selecting a preset number of Sipunculus nudus from different groups each month to test the sperm or egg development status, and then adjusting the feeding of Sipunculus nudus from different groups according to the development test results so that the development speed of Sipunculus nudus from different groups meets the preset synchronization requirements.

[0018] As a preferred implementation method, solution S04 preferably includes:

[0019] After draining the water used to cultivate Sipunculus nudus, allow it to dry for 2-3 days. Then, submerge the cultivation area with seawater at a salinity of 15 ppt and a temperature of 30°C for 2-3 hours. Drain the water again, and then reduce the salinity by 2 ppt and increase the temperature by 1°C each time. Repeat this process 2-3 times to physiologically stimulate the cultivated Sipunculus nudus to release eggs or sperm. During each soaking, check the sperm and egg release of the Sipunculus nudus in the water. Once sperm or eggs of Sipunculus nudus are found in the water, collect them through a mesh screen of a predetermined density for artificial insemination.

[0020] Based on the above, the present invention also provides a method for breeding Sipunculus nudus, which includes the above-described method for synchronizing the maturation of Sipunculus nudus sperm and eggs.

[0021] By adopting the above technical solution, the present invention has the following beneficial effects compared with the prior art: The present solution ingeniously discloses a method for synchronizing the maturation of sperm and eggs of Sipunculus nudus. By simulating the ebb and flow of tides and light in the daily breeding environment of Sipunculus nudus, the Sipunculus nudus exhibits better growth and survival rates and egg production. In addition, the present solution provides physiological stimulation to Sipunculus nudus that have reached physiological maturity in terms of sperm and egg release. By repeatedly soaking and gradually changing the salinity and temperature of seawater, the present solution promotes egg and sperm production in Sipunculus nudus, while also ensuring a good hatching rate for fertilized eggs. The present solution method can be used in the artificial breeding of Sipunculus nudus, and it is not only reliable to implement but also has high reproductive efficiency and high survival rate. Detailed Implementation

[0022] The present invention will be further described in detail below with reference to the embodiments. It should be particularly noted that the following embodiments are for illustrative purposes only and do not limit the scope of the invention. Similarly, the following embodiments are only some, not all, embodiments of the present invention, and all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0023] Example 1

[0024] This embodiment provides a method for synchronizing the maturation of sperm and eggs of the sipuncula worm, which includes:

[0025] S01. In October, select palatable Sipunculus nudus that meet the preset health requirements (no surface blemishes or lesions) and weigh between 3 and 4 g, and classify them by sex.

[0026] S02. The *Sipunculus nudus* were divided into groups of 40, with a female-to-male ratio of 3:1, and then transferred to culture soil for rearing. The culture soil was prepared at a concentration of 35 g / m³. 2 Add fermented chicken manure as base fertilizer;

[0027] S03. The breeding environment of *Sipunculus nudus* was controlled according to a preset light duration of 10.5 hours per day, a salinity of 24±1 ppt, and a pH of 6.5±0.2 to mimic the natural ecology. Simultaneously, the water in the breeding environment was added and removed twice daily to simulate tidal changes. Furthermore, diatoms were added to the water in the breeding environment every 15 days at a dosage of 10 g / m³. 2 In addition, a predetermined number of Sipunculus nudus from different groups are randomly selected each month to test the sperm or egg development. Then, based on the development test results, the feeding of Sipunculus nudus from different groups is adjusted so that the development speed of Sipunculus nudus from different groups meets the predetermined synchronization requirements.

[0028] S04. When the culture reaches June of the following year, drain the water used to culture Sipunculus nudus and let it dry for 2 days. Then, submerge the culture area of ​​Sipunculus nudus with seawater with a salinity of 15 ppt and a temperature of 30 ℃ for 2.5 hours. After that, drain the water again. Then, each time, decrease the salinity of the seawater by 2 ppt and increase the temperature by 1 ℃, and repeat the soaking twice to physiologically stimulate the cultured Sipunculus nudus to release eggs or sperm. During the soaking period, check the sperm and egg release of Sipunculus nudus in the water each time. When the sperm or eggs of Sipunculus nudus are found in the water, collect the sperm and eggs through a mesh of preset density for artificial insemination.

[0029] Example 2

[0030] This embodiment provides a method for synchronizing the maturation of sperm and eggs of the sipuncula worm, which includes:

[0031] S01. In October, select palatable Sipunculus nudus that meet the preset health requirements (no surface blemishes or lesions) and weigh between 3 and 4 g, and classify them by sex.

[0032] S02. The *Sipunculus nudus* were divided into groups of 40, with a female-to-male ratio of 3:1, and then transferred to culture soil for rearing. The culture soil was prepared at a concentration of 35 g / m³. 2 Add fermented chicken manure as base fertilizer;

[0033] S03. The breeding environment of *Sipunculus nudus* was controlled according to a preset light duration of 10.5 hours per day, a salinity of 24±1 ppt, and a pH of 6.5±0.2 to mimic the natural ecology. Simultaneously, the breeding environment was subjected to two water inflows and outflows daily to simulate tidal changes. Diatoms were also added to the water daily at a rate of 30 g / m³. 2In addition, a predetermined number of Sipunculus nudus from different groups are randomly selected each month to test the sperm or egg development. Then, based on the development test results, the feeding of Sipunculus nudus from different groups is adjusted so that the development speed of Sipunculus nudus from different groups meets the predetermined synchronization requirements.

[0034] S04. When the culture reaches June of the following year, drain the water used to culture Sipunculus nudus and let it dry for 2 days. Then, submerge the culture area of ​​Sipunculus nudus with seawater with a salinity of 15 ppt and a temperature of 30 ℃ for 2.5 hours. After that, drain the water again. Then, each time, decrease the salinity of the seawater by 2 ppt and increase the temperature by 1 ℃, and repeat the soaking twice to physiologically stimulate the cultured Sipunculus nudus to release eggs or sperm. During the soaking period, check the sperm and egg release of Sipunculus nudus in the water each time. When the sperm or eggs of Sipunculus nudus are found in the water, collect the sperm and eggs through a mesh of preset density for artificial insemination.

[0035] Example 3

[0036] This embodiment provides a method for synchronizing the maturation of sperm and eggs of the sipuncula worm, which includes:

[0037] S01. In October, select palatable Sipunculus nudus that meet the preset health requirements (no surface blemishes or lesions) and weigh between 3 and 4 g, and classify them by sex.

[0038] S02. The *Sipunculus nudus* were divided into groups of 40, with a female-to-male ratio of 3:1, and then transferred to culture soil for rearing. The culture soil was prepared at a concentration of 35 g / m³. 2 Add fermented chicken manure as base fertilizer;

[0039] S03. The breeding environment of *Sipunculus nudus* was controlled according to a preset light duration of 10.5 hours per day, a salinity of 24±1 ppt, and a pH of 6.5±0.2 to mimic the natural ecology. Simultaneously, the water in the breeding environment was added and removed twice daily to simulate tidal changes. Furthermore, diatoms were added to the water in the breeding environment every 15 days at a dosage of 50 g / m³. 2 In addition, a predetermined number of Sipunculus nudus from different groups are randomly selected each month to test the sperm or egg development. Then, based on the development test results, the feeding of Sipunculus nudus from different groups is adjusted so that the development speed of Sipunculus nudus from different groups meets the predetermined synchronization requirements.

[0040] S04. When the culture reaches June of the following year, drain the water used to culture Sipunculus nudus and let it dry for 2 days. Then, submerge the culture area of ​​Sipunculus nudus with seawater with a salinity of 15 ppt and a temperature of 30 ℃ for 2.5 hours. After that, drain the water again. Then, each time, decrease the salinity of the seawater by 2 ppt and increase the temperature by 1 ℃, and repeat the soaking twice to physiologically stimulate the cultured Sipunculus nudus to release eggs or sperm. During the soaking period, check the sperm and egg release of Sipunculus nudus in the water each time. When the sperm or eggs of Sipunculus nudus are found in the water, collect the sperm and eggs through a mesh of preset density for artificial insemination.

[0041] Example 4

[0042] This embodiment provides a method for synchronizing the maturation of sperm and eggs of the sipuncula worm, which includes:

[0043] S01. In October and May, select palatable Sipunculus nudus that meet the preset health requirements (no surface blemishes or lesions) and weigh between 3 and 4 g, and classify them by sex.

[0044] S02. The *Sipunculus nudus* were divided into groups of 40, with a female-to-male ratio of 3:1, and then transferred to culture soil for rearing. The culture soil was prepared at a concentration of 35 g / m³. 2 Add fermented chicken manure as base fertilizer;

[0045] S03. The breeding environment of *Sipunculus nudus* was controlled according to a preset light duration of 10.5 hours per day, a salinity of 24±1 ppt, and a pH of 6.5±0.2 to mimic the natural ecology. Simultaneously, the water in the breeding environment was added and removed twice daily to simulate tidal changes. Furthermore, diatoms were added to the water in the breeding environment every 15 days at a dosage of 50 g / m³. 2 In addition, a predetermined number of Sipunculus nudus from different groups are randomly selected each month to test the sperm or egg development. Then, based on the development test results, the feeding of Sipunculus nudus from different groups is adjusted so that the development speed of Sipunculus nudus from different groups meets the predetermined synchronization requirements.

[0046] S04. When the culture reaches June of the following year, drain the water used to culture Sipunculus nudus and let it dry for 2 days. Then, submerge the culture area of ​​Sipunculus nudus with seawater with a salinity of 15 ppt and a temperature of 30 ℃ for 2.5 hours. After that, drain the water again. Then, each time, decrease the salinity of the seawater by 2 ppt and increase the temperature by 1 ℃, and repeat the soaking twice to physiologically stimulate the cultured Sipunculus nudus to release eggs or sperm. During the soaking period, check the sperm and egg release of Sipunculus nudus in the water each time. When the sperm or eggs of Sipunculus nudus are found in the water, collect the sperm and eggs through a mesh of preset density for artificial insemination.

[0047] Comparative Example 1

[0048] This embodiment provides a method for synchronizing the maturation of sperm and eggs of the sipuncula worm, which includes:

[0049] S01. In October, select palatable Sipunculus nudus that meet the preset health requirements (no surface blemishes or lesions) and weigh between 3 and 4 g, and classify them by sex.

[0050] S02. The *Sipunculus nudus* were divided into groups of 40, with a female-to-male ratio of 3:1, and then transferred to culture soil for rearing. The culture soil was prepared at a concentration of 35 g / m³. 2 Add fermented chicken manure as base fertilizer;

[0051] S03. The breeding environment of *Sipunculus nudus* was controlled according to a preset light duration of 9–12 hours per day, a salinity of 24 ± 1 ppt, and a pH of 6.5 ± 0.2. The water in the breeding environment was incubated and drained once every 3 days. Diatoms were also added to the water in the breeding environment every 15 days at a rate of 50 g / m³. 2 In addition, a predetermined number of Sipunculus nudus from different groups are randomly selected each month to test the sperm or egg development. Then, based on the development test results, the feeding of Sipunculus nudus from different groups is adjusted so that the development speed of Sipunculus nudus from different groups meets the predetermined synchronization requirements.

[0052] S04. When the culture reaches June of the following year, drain the water used to culture Sipunculus nudus and let it dry for 2 days. Then, submerge the culture area of ​​Sipunculus nudus with seawater with a salinity of 15 ppt and a temperature of 30 ℃ for 2.5 hours. After that, drain the water again. Then, each time, decrease the salinity of the seawater by 2 ppt and increase the temperature by 1 ℃, and repeat the soaking twice to physiologically stimulate the cultured Sipunculus nudus to release eggs or sperm. During the soaking period, check the sperm and egg release of Sipunculus nudus in the water each time. When the sperm or eggs of Sipunculus nudus are found in the water, collect the sperm and eggs through a mesh of preset density for artificial insemination.

[0053] Comparative Example 2

[0054] This embodiment provides a method for synchronizing the maturation of sperm and eggs of the sipuncula worm, which includes:

[0055] S01. In October, select palatable Sipunculus nudus that meet the preset health requirements (no surface blemishes or lesions) and weigh between 3 and 4 g, and classify them by sex.

[0056] S02. The *Sipunculus nudus* were divided into groups of 40, with a female-to-male ratio of 3:1, and then transferred to culture soil for rearing. The culture soil was prepared at a concentration of 35 g / m³. 2 Add fermented chicken manure as base fertilizer;

[0057] S03. The breeding environment of *Sipunculus nudus* was controlled according to a preset light duration of 10.5 hours per day, a salinity of 24±1 ppt, and a pH of 6.5±0.2 to mimic the natural ecology. Simultaneously, the water in the breeding environment was added and removed twice daily to simulate tidal changes. Furthermore, diatoms were added to the water in the breeding environment every 15 days at a dosage of 50 g / m³. 2 In addition, a predetermined number of Sipunculus nudus from different groups are randomly selected each month to test the sperm or egg development. Then, based on the development test results, the feeding of Sipunculus nudus from different groups is adjusted so that the development speed of Sipunculus nudus from different groups meets the predetermined synchronization requirements.

[0058] S04. When the culture reaches June of the following year, drain the water used to culture Sipunculus nudus and let it dry for 2 days. Then, submerge the culture area of ​​Sipunculus nudus with seawater with a salinity of 15 ppt and a temperature of 30 ℃ for 2.5 hours. Repeat the soaking process twice to physiologically stimulate the cultured Sipunculus nudus to release eggs or sperm. During the soaking period, check the sperm and egg release of Sipunculus nudus in the water. When sperm or eggs of Sipunculus nudus are found in the water, collect the sperm and eggs through a mesh of preset density for artificial insemination.

[0059] In Examples 1-4 and Comparative Example 2, during a single 12-hour simulated high and low tide, the water intake maintained immersion in the aquaculture environment for 5 hours, semi-immersion in the culture environment for 1 hour, and dehydration for 6 hours. That is, in one day, the aquaculture environment was immersed in water for 10 hours, semi-immersion for 2 hours, and dehydration for 12 hours.

[0060] In addition, in S03, the feeding of different groups of *Sipunculus nudus* was adjusted according to the developmental test results to ensure that the developmental rate of different groups of *Sipunculus nudus* met the preset synchronization requirements. The method was to adjust the feeding of different groups of *Sipunculus nudus* at ±5 g / m 2 Within a certain range, the amount of diatoms added to different groups of Sipunculus nudus in each cycle can be adjusted. That is, when the weight or development of individual groups is slower, the amount added can be slightly increased, and when they tend to be average, the amount added can be restored to the normal setting.

[0061] Comparison data

[0062] To facilitate comparison of the above treatment methods and determine the optimal breeding strategy, this method extracts 10 kg of Sipunculus nudus parent organisms (several groups of Sipunculus nudus parent organisms totaling 10 kg) from Examples 1-4 and Examples 1-2 respectively. The survival rate of these parent organisms is counted, the number of eggs obtained is counted, and the hatching rate of fertilized eggs after artificial insemination is statistically analyzed. The results are shown in the table below:

[0063] Table 1 Comparison Data

[0064]

[0065] The results above show that each embodiment in this scheme has good technical effects. Among them, embodiment 3 is the best overall. In addition, in embodiments 1 and 2, because the amount of diatoms fed to Sipunculus nudus is relatively small, the number of eggs laid by Sipunculus nudus may decrease in the later stages of development due to the small amount of feed added. In addition, the amount of feed added regularly also affects the survival rate of Sipunculus nudus. In comparison with Comparative Example 1, it can be seen that Comparative Example 1 does not simulate the effect of tides on the growth of Sipunculus nudus. Survival and egg production are significantly affected, suggesting that simulating the wild environment and improving food supply are beneficial for the artificial breeding of Sipunculus nudus. Furthermore, Comparative Example 2 shows that multiple immersions and gradient changes in seawater salinity and temperature, as described in this scheme, promote egg production in Sipunculus nudus. In summary, this method is applicable to the artificial breeding of Sipunculus nudus, demonstrating reliable implementation, high reproductive efficiency, and high survival rate.

[0066] The above description is only a part of the embodiments of the present invention and does not limit the scope of protection of the present invention. Any equivalent device or equivalent process transformation made based on the content of the present invention specification, or direct or indirect application in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A method for the synchronized maturation of sperm and eggs of the palatable Sipunculus nudus, characterized in that, It includes: S01. Select healthy, palatable Sipunculus worms weighing 3-4 g and classify them by sex. S02. Every October, 40 Sipunculus nudus are grouped together, with a male-to-female ratio of 3:1, and then transferred to culture soil for breeding. S03. Light intensity was controlled in the breeding environment of *Sipunculus nudus* at a duration of 9–12 hours per day. Water quality was controlled at a salinity of 24 ± 1 ppt and a pH of 6.5 ± 0.

2. Diatomaceous earth was added to the water every 15 days at a rate of 10–50 g / m³. 2 To mimic the natural ecosystem, the breeding environment of Sipunculus nudus is irrigated and drained twice a day to simulate the ebb and flow of tides. S04. When the culture reaches June of the following year, drain the water used to culture Sipunculus nudus and let it dry for 2-3 days. Then, submerge the culture area of ​​Sipunculus nudus with seawater with a salinity of 15 ppt and a temperature of 30 ℃ for 2-3 hours. After soaking, drain the water again. Then, each time, decrease the salinity of the seawater by 2 ppt and increase the temperature by 1 ℃, and repeat the soaking 2-3 times to achieve physiological stimulation of the cultured Sipunculus nudus to induce egg or sperm release. During the soaking period, check the sperm and egg release of Sipunculus nudus in the water each time. When the sperm or eggs of Sipunculus nudus are found in the water, collect the sperm and eggs through a mesh for artificial insemination. S03 further includes: randomly sampling different groups of *Sipunculus nudus* each month, testing the sperm or egg development of the *Sipunculus nudus*, and then adjusting the feeding of different groups of *Sipunculus nudus* based on the development test results to ensure that the development speed of different groups of *Sipunculus nudus* meets the synchronization requirements, including: within ±5 g / m 2 Within the range, adjust the amount of diatoms added to different groups of Sipunculus nudus for each cycle. When the development of individual groups is lagging, increase the amount added. When they tend to be average, restore the normal amount added.

2. The method for synchronizing the maturation of sperm and eggs of Sipunculus nudus as described in claim 1, characterized in that, In S02, the culture soil contains 15–65 g / m³ 2 Add fermented chicken manure as base fertilizer.

3. The application of the method for synchronizing the maturation of sperm and eggs of Sipunculus nudus as described in claim 1 or 2 in the cultivation of Sipunculus nudus.

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