Artificial breeding method of scleractinian corals

Abstract

The application provides an artificial cultivation method of hard coral, and particularly relates to the technical field of coral cultivation. The artificial cultivation method comprises the following steps: constructing a water body environment with strong light and oligotrophic in a controllable temperature artificial cultivation water body; moving hard coral into the water body environment for ecological adaptation; after the hard coral is adapted, adding trace element additives in the water body environment, and establishing a periodic supplement mechanism for 6-9 days after the first addition; and adopting the Berlin system to realize continuous circulation treatment of the water body during cultivation. The method realizes organic integration of photochemical regulation, nutrition limitation strategy and an engineering circulation system, improves the growth condition of ornamental hard coral, improves the immune capacity of the coral, and effectively improves the survival rate and growth rate of the artificial cultivation coral. The technical advantage enables the method to be widely applied to the cultivation of ornamental hard coral in the aquarium market, and the method has wide market prospect and application potential.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of coral cultivation, in particular to an artificial cultivation method of hard coral. BACKGROUND

[0002] The coral reef ecosystem is known as the "tropical rainforest in the sea", and is one of the most complex and productive ecosystems on earth with the richest biodiversity, which has made a great contribution to the prosperity of marine fishery resources. In addition, corals are loved by humans for their colorful appearance and have a high ornamental value. However, with the intensification of the greenhouse effect and the influence of human activities such as wastewater discharge into the sea, corals naturally growing in the ocean are facing severe survival challenges. At the same time, with the rapid economic development and the continuous improvement of people's living standards, the demand for cultured ornamental corals is also growing. Therefore, improving the artificial cultivation technology of corals not only helps to promote the growth of corals, protect the coral reef ecosystem and maintain biodiversity, but also meets the market demand for ornamental corals, which is of great significance.

[0003] In the current aquarium market, ornamental corals often suffer a sharp decline in growth status after a series of processes such as sales, packaging and transportation. In addition, the cultivation environment provided by the existing artificial cultivation technology of corals is usually difficult to maintain for a long time in the most suitable conditions for the growth of corals, which has greatly restricted the growth of corals. In view of this, there is an urgent need for a cultivation technology and method that can effectively improve the growth status of artificially cultivated corals.

[0004] In view of the above, the present application is proposed. SUMMARY

[0005] The present application aims to provide an artificial cultivation method of hard coral, which aims to solve at least one of the above technical problems in the prior art.

[0006] In order to achieve the above-mentioned purpose of the present application, the following technical solutions are adopted:

[0007] The first aspect of the present application provides an artificial cultivation method of hard coral, comprising the following steps: constructing a water body environment with strong light and oligotrophic in a controllable temperature artificial cultivation water body; moving the hard coral into the water body environment for ecological adaptation; after the hard coral is adapted, adding a trace element additive in the water body environment, and establishing a periodic replenishment mechanism for 6-9 days after the first addition; and using the Berlin system to realize the continuous circulation treatment of the water body during the cultivation process.

[0008] Further, in the strong light and oligotrophic water environment, the wavelength of the light is in the blue-violet light section, the light intensity is 20000-24000k, the light period is 8-12 hours, and the remaining time is the dark period.

[0009] Preferably, in the water environment, the temperature is 24-25℃, the salinity is 25-29‰, and the pH is 8.1-8.3.

[0010] Preferably, in the water environment, the concentration of bicarbonate is 6.7-7.7, the calcium ion content is 410-440ppm, the magnesium ion content is 1380-1420ppm, the phosphate ion content is 0.01-0.04ppm, and the nitrate ion content is 1-2ppm.

[0011] Further, the trace element additive is composed of cationic trace element additives and anionic trace element additives.

[0012] The cationic trace element additive includes SrCl2, Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2, LiCl, and distilled water.

[0013] The anionic trace element additive includes NaBr, H3BO3, Na2MoO4, KI, KF, Na2CrO4, and distilled water.

[0014] Further, the cationic trace element additive includes SrCl2 70-150 parts, Fe-EDTA 5-15 parts, Mn-EDTA 1-2 parts, Zn-EDTA 0.5-1 part, Ni-EDTA 0.01-0.05 part, BaCl2 0.5-1 part, CuCl2 0.1-0.5 part, CoCl2 0.001-0.01 part, LiCl 1-1.5 parts, and distilled water 1000 parts, according to the weight fraction.

[0015] Further, the anionic trace element additive includes NaBr 100-150 parts, H3BO3 20-30 parts, Na2MoO4 0.4-1 part, KI 1-1.5 parts, KF 6-20 parts, Na2CrO4 0.001-0.005 parts, and distilled water 1000 parts, according to the weight fraction.

[0016] Further, the preparation method of the trace element additive includes the following steps:

[0017] A. Add SrCl2, Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2 and LiCl into distilled water and mix them evenly until a clear solution is obtained to obtain a cation trace element additive;

[0018] B. Add NaBr, H3BO3, Na2MoO4, KI, KF and Na2CrO4 into distilled water and mix them evenly until a clear solution is obtained to obtain an anion trace element additive.

[0019] Further, the ecological adaptation time is 2-4 weeks.

[0020] Further, the hard coral includes antler coral.

[0021] Preferably, the Berlin system includes live rock, protein separator, water flow system and light system.

[0022] Further, the addition amount of the trace element additive is 0.1-0.2 g / kg water.

[0023] Preferably, when periodic replenishment is performed, the addition amount of the trace element additive is 0.1-0.2 g / kg water.

[0024] Further, the time interval between the addition of the cation trace element additive and the anion trace element additive is greater than 1 hour.

[0025] Compared with the prior art, the present application has at least the following beneficial effects:

[0026] The artificial cultivation method provided by the present application effectively inhibits the competitive proliferation of large algae under the optimization of environmental parameters in a strong light and oligotrophic system, while maintaining the photosynthetic efficiency of zooxanthellae, thereby promoting the positive accumulation of energy of corals. The method combines the Berlin circulation system to realize efficient water circulation, and avoids the risk of ammonia nitrogen toxicity through precise regulation of water quality. By periodically replenishing trace elements, the calcification rate of corals is significantly improved, the density of zooxanthellae is stabilized, the symbiotic system is strengthened, and the stress resistance of corals is significantly enhanced. The method successfully breaks through the calcification bottleneck in artificial cultivation of hard corals through the organic integration of photochemical regulation, nutrient limitation strategy and engineered circulation system, providing a reliable technical path for coral conservation and marine ecological restoration. While improving the growth conditions of ornamental hard corals, the method significantly improves the immune capacity of corals, effectively improves the survival rate and growth rate of artificial cultivation corals. Its technical advantages enable it to be widely applied to the cultivation of ornamental hard corals in the aquarium market, and has broad market prospects and application potential. BRIEF DESCRIPTION OF DRAWINGS

[0027] In order to more clearly illustrate the technical solutions in the specific embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the specific embodiments or prior art description. Obviously, the drawings described below are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative effort on the basis of these drawings.

[0028] Figure 1 The figure for detecting the change of wet weight of corals is shown in the following table:

[0029] Figure 2 The figure for detecting the content of calcium and magnesium elements in corals is shown in the following table:

[0030] Figure 3 The figure for detecting the content of crude protein in corals is shown in the following table:

[0031] Figure 4 The figure for detecting the immune ability of corals is shown in the following table. DETAILED DESCRIPTION

[0032] In order to make the objects, technical solutions and advantages of the present application clearer, the following will combine the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments.

[0033] In the following, the terms "include", "have", and their synonymous words used in various embodiments of the present application are only intended to represent specific features, numbers, steps, operations, elements, components, or combinations of the foregoing, and should not be understood as first excluding the existence or possibility of adding one or more features, numbers, steps, operations, elements, components, or combinations of the foregoing.

[0034] The first aspect of the present application provides an artificial breeding method of hard coral, comprising the following steps: constructing a water body environment with strong light and oligotrophic in a controllable temperature artificial breeding water body; moving hard coral into the water body environment for ecological adaptation; after the hard coral is adapted, adding trace element additives in the water body environment, and establishing a periodic supplement mechanism for 6-9 days after the first addition; and adopting the Berlin system to realize the continuous circulation treatment of the water body during the breeding process.

[0035] The artificial breeding method provided by the present application effectively inhibits the competitive proliferation of large algae by optimizing environmental parameters in a strong light and oligotrophic system, while maintaining the photosynthetic efficiency of zooxanthellae, thereby promoting the positive energy accumulation of corals. The method combines the Berlin circulation system to achieve efficient water circulation, and avoids the risk of ammonia nitrogen toxicity by precisely regulating water quality. By periodically supplementing trace elements, the calcification rate of corals is significantly improved, the density of zooxanthellae is stabilized, the symbiotic system is strengthened, and the stress resistance of corals is significantly enhanced. The method successfully breaks through the calcification bottleneck in artificial breeding of scleractinian corals through the organic integration of photochemical regulation, nutrient limitation strategy and engineered circulation system, providing a reliable technical path for coral conservation and marine ecological restoration. While improving the growth conditions of ornamental scleractinian corals, the method significantly improves the immune capacity of corals, effectively improves the survival rate and growth rate of artificially bred corals. Its technical advantages enable it to be widely applied to the cultivation of ornamental scleractinian corals in the aquarium market, and has broad market prospects and application potential.

[0036] Further, in the strong light and oligotrophic water environment, the wavelength of light is in the blue-violet light segment, the light intensity is 20000-24000k, and the light cycle is 8-12 hours of light period and the remaining time is dark cycle.

[0037] Typically but not limitedly, the light intensity may be, for example, 20000k, 21000k, 22000k, 23000k or 24000k, or any value within the range of 20000k-24000k; the light cycle may be, for example, 8 hours, 9 hours, 10 hours, 11 hours or 12 hours, or any value within the range of 8-12 hours; and the dark cycle is the remaining time, i.e. 16 hours, 15 hours, 14 hours, 13 hours or 12 hours, or any value within the range of 12-16 hours.

[0038] When writing the light cycle, "10L / 14D" means 10 hours of light period to activate the photosynthetic system of coral symbiotic algae (zooxanthellae); 14 hours of dark cycle to simulate natural day-night rhythm and promote metabolic balance of corals.

[0039] Preferably, in the water environment, the temperature is 24-25℃, the salinity is 25-29‰, and the pH is 8.1-8.3.

[0040] Typically but not limitedly, the temperature in the water environment may be, for example, 24℃, 24.5℃, 25℃, or any value within the range of 24-25℃; the salinity may be, for example, 25‰, 26‰, 27‰, 28‰, 29‰, or any value within the range of 25-29‰; and the pH may be, for example, 8.1, 8.2, 8.3, or any value within the range of 8.1-8.3.

[0041] Preferably, the concentration of bicarbonate in the water environment is 6.7-7.7, the calcium ion content is 410-440 ppm, the magnesium ion content is 1380-1420 ppm, the phosphate ion content is 0.01-0.04 ppm, and the nitrate ion content is 1-2 ppm.

[0042] Typically but not limitedly, the concentration of bicarbonate may be, for example, 6.7, 7.0, 7.3, 7.7, or any value within the range of 6.7-7.7; the calcium ion content may be, for example, 410 ppm, 420 ppm, 430 ppm, 440 ppm, or any value within the range of 410-440 ppm; the magnesium ion content may be, for example, 1380 ppm, 1400 ppm, 1420 ppm, or any value within the range of 1380-1420 ppm; the phosphate ion content may be, for example, 0.01 ppm, 0.02 ppm, 0.03 ppm, 0.04 ppm, or any value within the range of 0.01-0.04 ppm; and the nitrate ion content may be, for example, 1 ppm, 1.5 ppm, 2 ppm, or any value within the range of 1-2 ppm. The concentration of bicarbonate is expressed using the KH value.

[0043] The present application provides sufficient light energy for the photosynthesis of coral symbiotic algae by setting up strong light culture conditions, thereby providing energy and material basis for the growth, development and reproduction of corals. At the same time, strong light conditions can promote the synthesis of pigments in corals, meet the market demand of artificial breeding of ornamental corals, and help them better adapt to artificial breeding environment by adjusting their physiological rhythms. In addition, the present application simulates the natural growth state of corals in seawater by setting up oligotrophic culture conditions, maintains their original physiological characteristics and ecological functions. This condition can effectively prevent harmful algae from over-reproduction in eutrophic environment, reduce the risk of coral diseases and competitive pressure, while maintaining the balance between corals and symbiotic algae, improving the stability and efficiency of the symbiotic system. Through the synergistic effect of strong light and oligotrophic conditions, the present application provides a scientific basis for the healthy growth of corals and the optimization of symbiotic systems, and lays a foundation for the sustainable development of artificial breeding of corals.

[0044] Further, the trace element additive is composed of a cationic trace element additive and an anionic trace element additive.

[0045] The cationic trace element additive includes SrCl2, Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2, LiCl and distilled water.

[0046] The anion trace element additive comprises NaBr, H3BO3, Na2MoO4, KI, KF, Na2CrO4 and distilled water.

[0047] The trace element additive contains various trace elements, has stable product state and high bioavailability of elements, and can effectively promote the growth of ornamental scleractinian corals and improve the immune capacity of the corals.

[0048] Further, the cation trace element additive comprises, in parts by weight, 70-150 parts of SrCl2, 5-15 parts of Fe-EDTA, 1-2 parts of Mn-EDTA, 0.5-1 part of Zn-EDTA, 0.01-0.05 part of Ni-EDTA, 0.5-1 part of BaCl2, 0.1-0.5 part of CuCl2, 0.001-0.01 part of CoCl2, 1-1.5 parts of LiCl and 1000 parts of distilled water.

[0049] Typically but not exclusively, the parts of SrCl2 in the cation trace element additive may be, for example, 70 parts, 80 parts, 90 parts, 100 parts, 110 parts, 120 parts, 130 parts, 140 parts, 150 parts, or any value within the range of 70-150 parts; the parts of Fe-EDTA may be, for example, 5 parts, 8 parts, 10 parts, 12 parts, 15 parts, or any value within the range of 5-15 parts; the parts of Mn-EDTA may be, for example, 1 part, 1.5 parts, 2 parts, or any value within the range of 1-2 parts; the parts of Zn-EDTA may be, for example, 0.5 part, 0.7 part, 1 part, or any value within the range of 0.5-1 part; the parts of Ni-EDTA may be, for example, 0.01 part, 0.02 part, 0.03 part, 0.04 part, 0.05 part, or any value within the range of 0.01-0.05 part; the parts of BaCl2 may be, for example, 0.5 part, 0.7 part, 1 part, or any value within the range of 0.5-1 part; the parts of CuCl2 may be, for example, 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, or any value within the range of 0.1-0.5 part; the parts of CoCl2 may be, for example, 0.001 part, 0.005 part, 0.01 part, or any value within the range of 0.001-0.01 part; and the parts of LiCl may be, for example, 1 part, 1.2 part, 1.5 part, or any value within the range of 1-1.5 part.

[0050] Further, the anion trace element additive comprises NaBr 100-150 parts, H3BO3 20-30 parts, Na2MoO4 0.4-1 part, KI 1-1.5 parts, KF 6-20 parts, Na2CrO4 0.001-0.005 parts and distilled water 1000 parts by weight.

[0051] Typically but not exclusively, the parts of NaBr in the anion trace element additive may, for example, be 100 parts, 120 parts, 130 parts, 140 parts, 150 parts, or any value within the range of 100-150 parts; the parts of H3BO3 may, for example, be 20 parts, 22 parts, 25 parts, 28 parts, 30 parts, or any value within the range of 20-30 parts; the parts of Na2MoO4 may, for example, be 0.4 parts, 0.5 parts, 0.7 parts, 1 part, or any value within the range of 0.4-1 part; the parts of KI may, for example, be 1 part, 1.2 parts, 1.5 parts, or any value within the range of 1-1.5 parts; the parts of KF may, for example, be 6 parts, 8 parts, 10 parts, 12 parts, 15 parts, 18 parts, 20 parts, or any value within the range of 6-20 parts; the parts of Na2CrO4 may, for example, be 0.001 parts, 0.002 parts, 0.003 parts, 0.004 parts, 0.005 parts, or any value within the range of 0.001-0.005 parts; and the parts of distilled water is 1000 parts.

[0052] Further, the preparation method of the trace element additive comprises the following steps:

[0053] A. adding SrCl2, Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2 and LiCl into distilled water to mix uniformly to a clear solution to obtain a cation trace element additive;

[0054] B. adding NaBr, H3BO3, Na2MoO4, KI, KF and Na2CrO4 into distilled water to mix uniformly to a clear solution to obtain an anion trace element additive.

[0055] Further, the ecological adaptation time is 2-4 weeks.

[0056] Further, the hard coral comprises an acropora.

[0057] Preferably, the Berlin system includes live rocks, protein separators, water flow systems, and lighting systems. The core components of the Berlin system include a large protein separator, one or more filter tanks for cultivating live rocks and live sand, and a sedimentation tank for sedimentation and collection of residues. The protein separator utilizes the principle of air bubbles to separate organic waste and proteins in the water, thereby reducing pollutants and ammonia nitrogen content in the water, maintaining clean and transparent water quality. Live rocks and live sand are key components of the Berlin system, which provide habitat for a large number of microorganisms that convert ammonia nitrogen into nitrite and further into nitrate through nitrification, thereby achieving biological purification of water quality. In addition, live rocks and live sand can provide abundant nutrients and minerals for organisms in the aquarium, promoting the growth of corals and other marine organisms.

[0058] Further, the addition amount of the trace element additive is 0.1-0.2 g / kg water.

[0059] By controlling the addition amount of the trace element additive to be within the range of 0.1-0.2 g / kg water, the demand of corals for trace elements at different growth stages can be accurately met, avoiding growth restriction due to insufficient addition or element concentration fluctuation caused by excessive addition, thereby ensuring healthy growth of corals.

[0060] In specific use, the addition amount of the trace element additive in each kg of water may be, for example, 0.1 g, 0.12 g, 0.14 g, 0.16 g, 0.18 g, or 0.2 g, or any value within the range of 0.1-0.2 g.

[0061] Preferably, when periodic replenishment is performed, the addition amount of the trace element additive is 0.1-0.2 g / kg water.

[0062] Further, the time interval between the addition of the cationic trace element additive and the anionic trace element additive is greater than 1 hour.

[0063] The present application requires the strategy of adding cationic and anionic trace element additives with a time interval of more than 1 hour, which significantly improves the efficiency of coral cultivation through multi-dimensional synergistic effect. This operation first avoids the precipitation reaction caused by the direct contact of cations (such as Sr 2+ , Ba 2+ ) and anions (such as MoO4 2- , CrO4 2- ), and optimizes the bioavailability of elements through the stability of EDTA chelate; secondly, it matches the absorption timing of the coral calcification layer and the symbiotic algae, improves the ion transmembrane transport efficiency, and at the same time maintains water quality stability, reduces the pH fluctuation range, and avoids I - oxidation loss.

[0064] The present application will be further described by specific examples and comparative examples, but it should be understood that these examples are only for a more detailed description and should not be construed as limiting the present application in any form. The raw materials used in the examples and comparative examples of the present application were prepared under conventional conditions or the conditions recommended by the manufacturer, unless otherwise specified. The reagents or instruments used were conventional products available on the market, unless otherwise specified.

[0065] Example 1

[0066] This example provides a method for artificial cultivation of hard coral, and the specific process is as follows:

[0067] 1. Cultivation environment: The coral was cultivated in a culture system composed of an aquarium with a size of 120 cm x 60 cm x 40 cm and a Berlin system. The water quality conditions were set as follows: temperature 24.5℃, salinity 27‰, pH 8.2, KH 7.2, Ca 2+ content 425 ppm, Mg 2+ content 1400 ppm, PO4 3- content 0.025 ppm, NO3 - content 1.5 ppm. The light conditions were set as follows: light cycle 10L / 14D, 24000k intensity of light provided by 80W LED blue-violet light. The water body was kept circulating and the water quality was monitored regularly, and the coral was placed in the culture system to carry out the experiment after the water quality reached the conditions suitable for coral growth.

[0068] 2. Coral selection: Three strains of healthy and healthy Acropora (Acropora, Acroporidae, Scleractinia, Hexacorallia, Cnidaria) with similar growth were selected and placed in the culture system described above. After the water quality conditions were stable and the coral adapted to the culture system for 14 days, the grouping experiment was carried out according to the experimental requirements.

[0069] 3. Addition of trace elements: In the aquarium, 0.2 g of trace element additive was added per kg of water, of which 0.1 g of cationic trace element additive and 0.1 g of anionic trace element additive.

[0070] In the cationic trace element additive, SrCl2·6H2O 120 parts, Fe-EDTA (chemical formula C 10 H 12 FeN2NaO8) 10 parts, Mn-EDTA (chemical formula C 10 H 12 MnN2NaO8) 1.5 parts, Zn-EDTA (chemical formula C 10 H 12ZnN2NaO8) 0.7 parts, BaCl2·2H2O 1 part, CuCl2·2H2O 0.1 parts, Ni-EDTA (chemical formula C 10 H 12 The solution is obtained by adding 0.03 parts of NiN2NaO8·xH2O, 0.007 parts of CoCl2·6H2O, and 1.2 parts of LiCl to 1L of distilled water and mixing until the solution is clear.

[0071] The anionic trace element additive is prepared by adding 130 parts of NaBr, 29 parts of H3BO3, 0.5 parts of Na2MoO4·2H2O, 1.2 parts of KI, 10 parts of KF·2H2O, and 0.004 parts of Na2CrO4·4H2O to 1L of distilled water and mixing until the solution is clear.

[0072] 4. Every 7 days, add 0.2g of trace element additive per kilogram of water to the aquarium, including 0.1g each of cationic and anionic trace element additives. The time interval between adding cationic and anionic trace element additives should be greater than 1 hour. Furthermore, replace 20% of the water in the aquarium every 7 days, and continue this process for at least 49 days.

[0073] Example 2

[0074] This embodiment provides a method for the artificial cultivation of bony corals, the specific process of which is as follows:

[0075] 1. Cultivation Environment: Corals were cultivated using a 120cm×60cm×40cm aquarium and a Berlin system. Water quality conditions were set as follows: temperature 24℃, salinity 25‰, pH 8.1, KH 6.7, Ca... 2+ Content 410ppm, Mg 2+ Content 1380ppm, PO4 3- Content 0.01ppm, NO3 - The concentration was 1 ppm. Lighting conditions were set as follows: 10L / 14D illumination cycle, with 20000k intensity light provided by an 80W LED blue-violet lamp. Water circulation was maintained, and water quality was monitored regularly. Corals were introduced for the experiment only after the water quality reached suitable conditions for coral growth.

[0076] 2. Coral selection: Select three healthy staghorn corals with similar growth (Acroporidae, Scleractinia, Hexacorallia, Cnidaria) and place them into the culture system described above. After the water quality stabilizes and the corals have adapted to the culture system for 14 days, conduct group experiments according to the experimental requirements.

[0077] 3. The addition of trace elements: In one of the aquariums, 0.2 g of trace element additives per kg of water was added, including 0.1 g of cationic trace element additives and 0.1 g of anionic trace element additives.

[0078] In the cationic trace element additives, 110 parts of SrCl2·6H2O, 5 parts of Fe-EDTA (chemical formula C 10 H 12 FeN2NaO8), 1 part of Mn-EDTA (chemical formula C 10 H 12 MnN2NaO8), 0.5 parts of Zn-EDTA (chemical formula C 10 H 12 ZnN2NaO8), 0.5 parts of BaCl2·2H2O, 0.1 parts of CuCl2·2H2O, 0.01 parts of Ni-EDTA (chemical formula C 10 H 12 NiN2NaO8·xH2O), 0.001 parts of CoCl2·6H2O, and 1 part of LiCl were added to 1 L of distilled water and mixed until the solution was clear.

[0079] In the anionic trace element additives, 100 parts of NaBr, 20 parts of H3BO3, 0.5 parts of Na2MoO4·2H2O, 1 part of KI, 10 parts of KF·2H2O, and 0.001 parts of Na2CrO4·4H2O were added to 1 L of distilled water and mixed until the solution was clear.

[0080] 4. Every 7 days, 0.2 g of trace element additives per kg of water was added to the aquarium, including 0.1 g of cationic trace element additives and 0.1 g of anionic trace element additives. The time interval between the cationic trace element additives and the anionic trace element additives was greater than 1 h. Additionally, 20% of the water in the aquarium was replaced every 7 days, and the cultivation was continued for at least 49 days.

[0081] Example 3

[0082] This example provides a method for artificial cultivation of hard coral, and the specific process is as follows:

[0083] 1. Cultivation environment: The cultivation system was composed of an aquarium with a size of 120 cm x 60 cm x 40 cm and a Berlin system for cultivating coral. The water quality conditions were set as follows: temperature 25℃, salinity 29‰, pH 8.3, KH 7.7, Ca 2+ content 440 ppm, Mg 2+ content 1420 ppm, PO4 3- content 0.04 ppm, NO3 -The content is 2 ppm. The light condition is set as: light cycle 10L / 14D, provided by 80W LED blue-violet light with 24000k intensity. Keep the water circulating and monitor the water quality regularly, and put the coral into the experiment after the water quality reaches the suitable condition for coral growth.

[0084] 2. Coral selection: Select 3 strains of similar and healthy Acropora (Acroporidae, Scleractinia, Hexacorallia, Cnidaria) and put them into the culture system described above. After the water quality is stable and the coral has adapted to the culture system for 14 days, the grouping experiment is carried out according to the experimental requirements.

[0085] 3. The addition condition of trace elements: In one of the aquariums, add 0.2g of trace element additives per kg of water, including 0.1g of cation trace element additives and 0.1g of anion trace element additives.

[0086] In the cation trace element additive, SrCl2·6H2O 150 parts, Fe-EDTA (chemical formula C 10 H 12 FeN2NaO8) 15 parts, Mn-EDTA (chemical formula C 10 H 12 MnN2NaO8) 2 parts, Zn-EDTA (chemical formula C 10 H 12 ZnN2NaO8) 1 part, BaCl2·2H2O 1 part, CuCl2·2H2O 0.5 part, Ni-EDTA (chemical formula C 10 H 12 NiN2NaO8·xH2O) 0.05 parts, CoCl2·6H2O 0.01 parts, LiCl 1.5 parts are added to 1L distilled water to mix until the solution is clear.

[0087] In the anion trace element additive, NaBr 150 parts, H3BO3 30 parts, Na2MoO4·2H2O 1 part, KI 1.5 parts, KF·2H2O 20 parts, Na2CrO4·4H2O 0.005 parts are added to 1L distilled water to mix until the solution is clear.

[0088] 4. Every 7 days, add 0.2g of trace element additives per kg of water in the aquarium, including 0.1g of cation trace element additives and 0.1g of anion trace element additives. The time interval between cation trace element additives and anion trace element additives is more than 1h. And change 20% of the water in the aquarium every 7 days, and continue to culture for at least 49 days.

[0089] Comparative Example 1

[0090] This comparative example provides a method for artificial cultivation of hard coral, the specific process is as follows:

[0091] 1. The cultivation environment is the same as that in Example 1.

[0092] 2. The coral selection is the same as that in Example 1.

[0093] 3. The addition condition of trace elements: only 0.1 g of cation trace element additive is added per kilogram of water in the aquarium.

[0094] The formula of the cation trace element additive is the same as that in Example 1.

[0095] 4. This step is the same as in Example 1.

[0096] Comparative Example 2

[0097] This comparative example provides a method for artificial cultivation of hard coral, the specific process is as follows:

[0098] 1. The cultivation environment is the same as that in Example 1.

[0099] 2. The coral selection is the same as that in Example 1.

[0100] 3. The addition condition of trace elements: only 0.1 g of anion trace element additive is added per kilogram of water in the aquarium.

[0101] The formula of the anion trace element additive is the same as that in Example 1.

[0102] 4. This step is the same as in Example 1.

[0103] Comparative Example 3

[0104] This comparative example provides a method for artificial cultivation of hard coral, the specific process is as follows:

[0105] 1. The cultivation environment is the same as that in Example 1.

[0106] 2. The coral selection is the same as that in Example 1.

[0107] 3. The addition condition of trace elements: no trace elements are added in the aquarium, and the natural growth state of the coral is maintained.

[0108] 4. This step is the same as in Example 1.

[0109] Comparative Example 4

[0110] This comparative example provides a method for artificial cultivation of hard coral, the specific process is as follows:

[0111] 1. Breeding environment: The coral was cultured in a culture system composed of an aquarium with a size of 120 cm x 60 cm x 40 cm and a Berlin system. The water quality conditions were set as follows: temperature 24.5°C, salinity 27‰, pH 8.2, KH 7.2, Ca 2+ content 425 ppm, Mg 2+ content 1400 ppm, PO4 3- content 0.025 ppm, NO3 - content 1.5 ppm. The water body was kept circulating and the water quality was monitored regularly. After the water quality reached the conditions suitable for coral growth, the coral was put into the culture system to carry out the experiment.

[0112] 2. Coral selection: Three strains of healthy and healthy Acropora (Acropora, Acroporidae, Scleractinia, Hexacorallia, Cnidaria) with similar growth were selected and put into the culture system described above. The culture system was not turned on. After the water quality conditions were stable and the coral was adapted to the culture system for 14 days, the grouping experiment was carried out according to the experimental requirements.

[0113] 3. The same as step 1 of Example 1.

[0114] 4. The same as step 1 of Example 1.

[0115] Test Example

[0116] 1. The coral obtained in the example (EG) and the comparative example (CG) was detected, which specifically included:

[0117] Wet weight change detection: The wet weight of the coral was weighed by an analytical balance on the 0th day and the 49th day of the experiment, and the increase and rate of the wet weight of the coral were calculated, and the results are shown in Table 1 and Figure 1 .

[0118] Element content detection: The calcium and magnesium element contents in the coral of the example and the comparative example were determined by ICP-OES / MS method. Appropriate amount of coral tissue sample was taken on a clean bench, and nitric acid was used for sample digestion. ICP-OES method was determined by inductively coupled plasma emission spectrometer, and the characteristic spectral line wavelength of the element was qualitative, and the spectral line signal intensity of the measured element was proportional to the element concentration for quantitative analysis. ICP-MS was determined by inductively coupled plasma mass spectrometer, and the element specific mass number (mass-to-charge ratio, m / z) was qualitative, and the intensity ratio of the mass spectrum signal of the measured element to the internal standard element was proportional to the concentration of the measured element for quantitative analysis. The detection results are shown in Table 2 and Figure 2 .

[0119] Crude protein content detection: Kjeldahl method was used to detect the crude protein content in the corals of the examples and the comparative examples. Appropriate amount of coral tissue sample was taken on a clean bench and moved into a digestion tube. Kjeldahl catalyst and concentrated sulfuric acid were added, and the sample was digested on a 400℃ digestion furnace for 1h. After cooling, the sample was taken out and diluted to volume, and then filtered or left to clarify for nitrogen determination. After preheating the nitrogen determination instrument, the sample was taken and injected into the Kjeldahl distillation tube, and the parameters were set to determine the sample. The results are shown in Table 3 and Figure 3

[0120] Immune ability detection: The activities of catalase (CAT), total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in the corals of the examples and the comparative examples were determined by using a kit. Appropriate amount of coral tissue sample was taken on a clean bench, and excess ammonium molybdate was added to stop the reaction of hydrogen peroxide decomposition by catalase. The remaining hydrogen peroxide reacted with ammonium molybdate to form a light yellow complex. The change in absorbance of the light yellow complex was determined by a microplate reader at 405nm to obtain the CAT detection result. Appropriate amount of coral tissue sample was taken on a clean bench, and an extraction solution was added to reduce Fe 3+ -TPTZ to generate blue Fe 2+ -TPTZ. The change in absorbance was determined by a microplate reader at 593nm to obtain the T-AOC detection result. Appropriate amount of coral tissue sample was taken on a clean bench, and an extraction solution was added to allow superoxide dismutase to scavenge superoxide anion and inhibit the reaction process of superoxide anion reducing nitrogen blue tetrazolium to generate blue methylene. The absorbance of the reaction solution was determined by a microplate reader at 560nm to obtain the SOD content. The detection results are shown in Table 4 and Figure 4

[0121] 2. Detection result analysis

[0122] 2.1. Analysis of wet weight change detection data

[0123] Table 1 Detection results of coral wet weight change

[0124]

[0125] As shown in Table 1 and Figure 1 After 49 days of culture according to the method of the present application, the average wet weight of the coral samples in the experimental examples was significantly increased, which was significantly higher than that of the comparative examples. Among them, the effect of experimental example 1 was the best, and the average wet weight of the coral increased by 1.34g, which increased by 103.08% within 49 days. In summary, the method of adding trace elements to culture ornamental scleractinian corals according to the present application can significantly promote the increase of the wet weight of the scleractinian corals.

[0126] 2.2. Analysis of element content detection data

[0127] ​​Table 2. Results of calcium and magnesium content detection in coral.

[0128]

[0129] As shown in Table 2 and Figure 2 As shown, after 49 days of cultivation according to the method described in this invention, the average contents of calcium and magnesium in the coral samples of the experimental examples were significantly higher than those of the comparative examples (p≤0.05). Specifically, the average contents of calcium and magnesium in the corals of Example 1 were the highest, at 27.096 g / kg and 0.46 g / kg, respectively; while the average contents of calcium and magnesium in the corals of Comparative Example 4 were the lowest, at 21.085 g / kg and 0.377 g / kg, respectively. Therefore, cultivating corals according to the method described in this invention can significantly enhance the absorption and utilization of calcium and magnesium in the water by corals, promoting the growth of bony corals.

[0130] 2.3 Analysis of Crude Protein Content Detection Data

[0131] Table 3. Results of crude protein content detection in corals

[0132] Crude protein (CP) average content (%) Example 1 1.93 Example 2 1.89 Example 3 1.88 Comparative Example 1 1.84 Comparative Example 2 1.83 Comparative Example 3 1.8 Comparative Example 4 1.69

[0133] As shown in Table 3 and Figure 3 As shown, after 49 days of cultivation according to the method described in this invention, the average crude protein content in the coral samples of the experimental examples was significantly higher than that of the comparative examples (p≤0.05). Among them, the average crude protein content in the corals of Example 1 was the highest, at 1.93%; while the average crude protein content in the corals of Comparative Example 4 was the lowest, at 1.69%. Therefore, the method of adding trace elements to cultivate ornamental bony corals described in this invention can improve the synthesis of proteins in corals and promote the growth of bony corals.

[0134] 2.4 Analysis of Immunity Detection Data

[0135] Table 4 Results of Coral Immunity Test

[0136]

[0137] As shown in Table 4 and Figure 4 As shown, after culturing for 49 days according to the method described in this invention, the average values ​​of CAT, T-AOC, and SOD in the coral samples of the experimental examples were significantly higher than the corresponding indicators in the comparative examples (p≤0.05). Among them, Example 1 showed the best results, with an average CAT activity of 49.392 U / g and an average T-AOC of 0.697 μmol Fe. 2+ / g, and the SOD activity is 127.665 U / g. In conclusion, the method for culturing the ornamental scleractinian coral by adding trace elements can effectively improve the immune ability of the coral, and further improve the survival rate of the coral.

[0138] Finally, it should be noted that the above-described embodiments are merely specific implementations of the present application, which are used to illustrate the technical solutions of the present application, but not to limit the same, and the protection scope of the present application is not limited thereto. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that any person skilled in the art can still modify or easily think of changes to the technical solutions recorded in the foregoing embodiments, or make equivalent replacements to some of the technical features, without departing from the technical scope disclosed by the present application. Such modifications, changes or replacements do not make the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application, and should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A method for artificial culturing of hard corals, characterized in that, Includes the following steps: A high-light, low-nutrient aquatic environment was created in a temperature-controlled artificial culture water body; and bony corals were transferred into the aquatic environment to undergo ecological adaptation. After the bony corals have adapted, trace element additives are added to the aquatic environment. After the first addition, a periodic replenishment mechanism of 6 to 9 days is established. During the cultivation process, the Berlin system is used to achieve continuous water circulation. In the aforementioned aquatic environment with strong light and low nutrients, the light wavelength is in the blue-violet range, the light intensity is 20,000~24,000k, the light cycle is 8~12 hours of light period, and the remaining time is a dark period; The aquatic environment has a temperature of 24-25℃, a salinity of 25-29‰, and a pH of 8.1-8.

3. In the aforementioned aquatic environment, the concentration of bicarbonate is 6.7~7.7, the calcium ion content is 410~440ppm, the magnesium ion content is 1380~1420ppm, the phosphate ion content is 0.01~0.04ppm, and the nitrate ion content is 1~2ppm. The trace element additive is composed of cationic trace element additives and anionic trace element additives; According to the weight percentages, the cationic trace element additive comprises 70-150 parts SrCl2, 5-15 parts Fe-EDTA, 1-2 parts Mn-EDTA, 0.5-1 parts Zn-EDTA, 0.01-0.05 parts Ni-EDTA, 0.5-1 parts BaCl2, 0.1-0.5 parts CuCl2, 0.001-0.01 parts CoCl2, 1-1.5 parts LiCl, and 1000 parts distilled water; According to the weight parts, the anionic trace element additive includes 100-150 parts of NaBr, 20-30 parts of H3BO3, 0.4-1 parts of Na2MoO4, 1-1.5 parts of KI, 6-20 parts of KF, 0.001-0.005 parts of Na2CrO4 and 1000 parts of distilled water; The initial addition amount of the trace element additive is 0.1~0.2 g / kg water; When periodic supplementation is performed, the amount of the trace element additive added is 0.1~0.2g / kg water; The time interval between adding cationic and anionic trace element additives is greater than 1 hour.

2. The artificial breeding method according to claim 1, characterized by, The preparation method of the trace element additive includes the following steps: A. Add SrCl2, Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2 and LiCl to distilled water and mix until a clear solution is obtained to obtain a cationic trace element additive; B. Add NaBr, H3BO3, Na2MoO4, KI, KF and Na2CrO4 to distilled water and mix thoroughly until a clear solution is obtained to obtain anionic trace element additives.

3. The artificial breeding method according to claim 1, characterized by, The ecological adaptation period is 2 to 4 weeks.

4. The artificial breeding method according to any one of claims 1 to 3, characterized in that, The bony corals include staghorn corals.

5. The artificial breeding method according to any one of claims 1 to 3, characterized in that, The Berlin system includes live rock, a protein separator, a water flow system, and a lighting system.

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