Production process and use method of macrobrachium rosenbergii fry breeding seawater formula

Abstract

The invention discloses a macrobrachium rosenbergii breeding seawater formula production process and a use method, and belongs to the technical field of aquaculture, the macrobrachium rosenbergii breeding seawater formula comprises sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA, the mass ratio of which is sequentially reduced; the composition further comprises a beta-alanine methyl ester derivative and isomaltitol. The beta-alanine methyl ester derivative contains a guanidyl structure. By adopting the macrobrachium rosenbergii fry breeding seawater formula, the technological process for preparing artificial seawater can be simplified, quality control is facilitated, the growth performance of macrobrachium rosenbergii can be improved, and the macrobrachium rosenbergii fry breeding seawater formula is suitable for application and popularization.

Description

Technical Field

[0001] The invention belongs to the technical field of aquaculture, and in particular relates to a production process and a use method of a seawater formula for breeding Macrobrachium rosenbergii seedlings. Background Art

[0002] Artificial sea water, also known as artificial sea water, refers to a mixed solution of salts used to feed marine animals or preserve the organs and tissues of marine animals to keep them in a normal state. Because the osmotic pressure of seawater can vary from region to region, artificial seawater also uses different osmotic concentrations according to different regions and marine animals.

[0003] There are few prefabricated seawater products on the market. When breeding and raising Macrobrachium rosenbergii, fisheries mostly purchase ingredients separately according to existing formulas and then prepare artificial seawater by themselves. This leads to problems such as multiple ingredients, complex raw material varieties, and unguaranteed ingredient quality. Moreover, in the preparation process, most of them require repeated trials before the appropriate ingredients can be selected. The preparation process is complicated, error-prone, and lacks quality control, which is not suitable for efficient production requirements.

[0004] The invention application with publication number CN105210935A relates to a simulated seawater for breeding Macrobrachium rosenbergii and a preparation method thereof. The main components of the simulated seawater preparation include NaCl, MgSO 4 , CaCl 2 , KCl, NaBr, H 3 BO 3 、Na 2 SiO 3 、Al 2 Cl 6 The preparation method includes harmless treatment of fresh water before the simulated seawater preparation and harmless treatment after the simulated seawater preparation. The application enhances the physique of seedlings, removes parasites, has good water quality, and is easy to prepare. Some ingredients are higher than those contained in natural seawater, such as Ca 2+ The purpose is to provide a good material basis for the seedlings when peeling, while adding trace elements to enhance the activity and physical fitness of the seedlings.

[0005] However, similar to existing seawater formulas, the ingredients of the simulated seawater often need to be self-prepared. The artificial seawater obtained from different batches varies greatly, and the material easily absorbs moisture, which is not only not conducive to packaging, but also will clump after being stacked for a long time, thus affecting transportation and being unfavorable for use. Summary of the invention

[0006] The purpose of the present invention is to provide a production process and a use method of a seawater formula for breeding Macrobrachium rosenbergii seedlings, which can simplify the process flow of configuring artificial seawater, is conducive to quality control, and can improve the growth performance of Macrobrachium rosenbergii, and is suitable for popularization and application.

[0007] The technical solution adopted by the present invention to achieve the above-mentioned purpose is: A seawater formula for breeding Macrobrachium rosenbergii seedlings, comprising sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA; Among them, sodium chloride accounts for 65-75 parts, magnesium sulfate accounts for 20-28 parts, anhydrous calcium chloride accounts for 1-3 parts, potassium chloride accounts for 0.5-2 parts, potassium bromide accounts for 0.1-0.3 parts, boric acid accounts for 0.5-1.5 parts, and EDTA accounts for 0.01-0.05 parts.

[0008] The method for using the above-mentioned seawater formula for raising Macrobrachium rosenbergii seedlings is to add 13-16 kilograms of the seawater formula per cubic meter of target seawater.

[0009] Preferably, 14.5 kg of the seawater formula is added per cubic meter of target seawater.

[0010] Furthermore, when using the seawater formula, anhydrous calcium chloride may be additionally added, and the amount of the additional anhydrous calcium chloride added is 0.03-0.06 kg per cubic meter of water.

[0011] By adopting the above-mentioned seawater formula for breeding Macrobrachium rosenbergii, the seawater formula can be directly used after being diluted, which simplifies the process of configuring artificial seawater in the fishery, ensures the identity of the artificial seawater obtained by multiple configurations, and realizes standardized quality control; and reduces the use of redundant ingredients, thereby reducing the cost of raw materials; the processing technology of the seawater formula for breeding Macrobrachium rosenbergii can greatly reduce the probability of materials absorbing water and becoming damp, avoid material agglomeration, and facilitate packaging and transportation; in the preparation of the seawater formula for breeding Macrobrachium rosenbergii, mother salt is first configured, and then the remaining components are mixed with it, which can avoid the generation of a large amount of dust during the mixing process, and can also reduce the probability of materials adhering to the inner wall of the mixer, thereby saving raw material costs and ensuring the quality of the artificial seawater obtained by adopting the seawater formula.

[0012] Furthermore, the seawater formula also includes a β-alanine methyl ester derivative and isomalt; the β-alanine methyl ester derivative contains a guanidine structure.

[0013] Preferably, in the seawater formula, the mass ratio of the β-alanine methyl ester derivative to isomalt is 1:0.5-1.5, and the mass ratio of the β-alanine methyl ester derivative to sodium chloride in the seawater formula is 1:30-120.

[0014] In the seawater formula, in addition to sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA, a certain amount of β-alanine methyl ester derivatives and isomalt are added. The seawater formula is used to prepare artificial seawater for the cultivation of Macrobrachium rosenbergii, which can not only improve the antibacterial properties, but also improve the survival rate, weight gain rate and crude protein content of Macrobrachium rosenbergii.

[0015] This may be because of the presence of guanidine groups in β-alanine methyl ester derivatives and the polyhydroxy structure in isomalt. The combination of the two can react with various pathogens that may exist in aquaculture water, such as Vibrio, Aeromonas, and Flavobacterium, thereby avoiding the infection of pathogens to Macrobrachium rosenbergii, thereby reducing the incidence of septicemia, ulcers, tail rot, enteritis, and vibriosis in Macrobrachium rosenbergii, thereby improving the survival rate of Macrobrachium rosenbergii. On the other hand, β-alanine methyl ester derivatives also have amino and imino structures, rich in N elements, and they themselves contain β-alanine ester groups, which can reduce the difficulty of contact with pathogens, thereby improving antibacterial properties. The addition of β-alanine methyl ester derivatives and isomalt to aquaculture water provides a healthy and good growth environment for Macrobrachium rosenbergii, and also helps to increase the weight gain rate of Macrobrachium rosenbergii. On the other hand, β-alanine methyl ester derivatives also contain amino and imino structures, rich in N elements, and they themselves contain β-alanine groups, which themselves also contribute to the growth of Macrobrachium rosenbergii and provide a source of N for protein synthesis of Macrobrachium rosenbergii. In addition, the presence of amino groups in β-alanine methyl ester derivatives and polyhydroxy structures in isomalt may promote the expression of related protein genes in Macrobrachium rosenbergii cells, thereby promoting the growth of Macrobrachium rosenbergii seedlings and increasing protein content.

[0016] According to one aspect of the present invention, a method for preparing a β-alanine methyl ester derivative is provided: Benzoyl isothiocyanate reacts with β-alanine methyl ester hydrochloride, and the resulting product reacts with NaOH to obtain a product with CH 3 I reaction, the obtained product is reacted with n-propylamine and N,N-diisopropylethylamine to obtain a β-alanine methyl ester derivative. The obtained β-alanine methyl ester derivative contains a guanidine structure.

[0017] Specifically, the preparation method of β-alanine methyl ester derivatives comprises the following steps: S1. Add benzoyl isothiocyanate to a DCM solution containing β-alanine methyl ester hydrochloride, stir the reaction, and then filter. The resulting filter residue is washed and dried. S2. The solid obtained in S1 was reacted with a NaOH solution, and then filtered, and the resulting residue was washed and dried; S3. The solid obtained in S2 is then mixed with CH 3I reaction, then filtering, washing and drying the obtained filter residue; S4. The solid obtained in S3, n-propylamine and N,N-diisopropylethylamine were added to anhydrous DMF and stirred for reaction; then, a NaCl solution was added thereto and dialyzed with deionized water to obtain a β-alanine methyl ester derivative.

[0018] More specifically, the preparation method of β-alanine methyl ester derivative is as follows: S1. Add benzoyl isothiocyanate to a DCM solution containing β-alanine methyl ester hydrochloride under ice bath conditions, stir at 25°C, 150r / min for 24h, then filter, wash the residue with DCM, and dry in vacuo at 55°C for 6h; S2. The solid obtained in S1 was dissolved in ethanol, and NaOH solution was added dropwise thereto. The mixture was stirred at 25°C and 150 r / min for 24 h, and then filtered. The residue was washed with ethanol and dried under vacuum at 55°C for 6 h. S3. The solid obtained in S2 was dissolved in ethanol and CH 3 I, 40°C, 150r / min stirring reaction for 48h, then filtered, the obtained filter residue was washed with ethanol, and vacuum dried at 55°C for 6h; S4. The solid obtained in S3, n-propylamine and N,N-diisopropylethylamine were added to anhydrous DMF, and the mixture was stirred at 60°C and 150 r / min for 72 h. Then, a NaCl solution was added thereto, and the mixture was dialyzed with deionized water to obtain a β-alanine methyl ester derivative.

[0019] Preferably, in S1, the molar ratio of benzoyl isothiocyanate to β-alanine methyl ester hydrochloride is 1-1.5:1.

[0020] Preferably, in S1, the mass volume ratio of β-alanine methyl ester hydrochloride to DCM is 1 g:60-120 mL.

[0021] Preferably, in S2, the mass volume ratio of the solid obtained in S1 to ethanol is 1 g:15-40 mL; Preferably, in S2, the concentration of the NaOH solution is 3.0-6.0 mol / L; Preferably, in S2, the mass volume ratio of the solid obtained in S1 to the NaOH solution is 1 g:1-5 mL.

[0022] Preferably, in S3, the mass volume ratio of the solid obtained in S2 to ethanol is 1 g:30-80 mL; Preferably, in S3, the solid obtained in S2 is reacted with CH 3 The mass ratio of I is 1:1-3.

[0023] Preferably, in S4, the mass concentration of the NaCl solution is 15-40%; the mass volume ratio of the solid obtained in S3 to the NaCl solution is 1 mg: 3-8 mL; Preferably, in S4, the MWCO of the membrane used for dialysis is 1000Da.

[0024] According to one aspect of the present invention, a seawater formula for raising Macrobrachium rosenbergii seedlings is provided, comprising sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA in descending order by weight; and also comprising a β-alanine methyl ester derivative, isomalt and 3-mercapto-2-butanone.

[0025] Preferably, the mass ratio of the β-alanine methyl ester derivative to isomalt is 1:0.5-1.5, the mass ratio of the β-alanine methyl ester derivative to sodium chloride in the seawater formula is 1:30-120; the mass ratio of the β-alanine methyl ester derivative to 3-mercapto-2-butanone is 1:0.5-1.5.

[0026] In the seawater formula for Macrobrachium rosenbergii seedlings, in addition to sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA, a certain amount of β-alanine methyl ester derivatives and isomalt are added, and a certain amount of 3-mercapto-2-butanone is further added, which can further improve the antibacterial properties of the obtained artificial seawater. This may be because 3-mercapto-2-butanone contains a thiol structure, which can further enhance its effect on pathogens by combining with the guanidine group in the β-alanine methyl ester derivative and the hydroxyl group in isomalt. In addition, the use of 3-mercapto-2-butanone in combination with β-alanine methyl ester derivatives and isomalt can further improve the survival rate, weight gain rate and crude protein content of Macrobrachium rosenbergii. This may be because the presence of thiol, carbonyl, etc. in 3-mercapto-2-butanone, together with the β-alanine ester group and guanidine group in β-alanine methyl ester derivatives, and / or the polyhydroxy structure in isomalt, helps to increase the expression of related protein genes in Macrobrachium rosenbergii, thereby promoting the growth of Macrobrachium rosenbergii seedlings and helping to increase the protein content.

[0027] Compared with the prior art, the present invention has the following beneficial effects: 1. Provide a formula for seawater for breeding Macrobrachium rosenbergii, which can simplify the process of configuring artificial seawater in a fishery and help to achieve standardized quality control of aquaculture water; the processing technology of the formula for seawater for breeding Macrobrachium rosenbergii can greatly reduce the probability of materials absorbing water and becoming damp, avoid material agglomeration, and facilitate packaging and transportation; in the preparation of the formula for seawater for breeding Macrobrachium rosenbergii, mother salt is first configured and then the remaining components are mixed therewith, which can avoid the generation of a large amount of dust during the mixing process and can also reduce the probability of materials adhering to the inner wall of the mixer, thereby saving raw material costs and ensuring the quality of artificial seawater obtained by using the seawater formula.

[0028] 2. Add a certain amount of β-alanine methyl ester derivatives and isomalt to the seawater formula, and use the seawater formula to prepare artificial seawater for the cultivation of Macrobrachium rosenbergii, which can not only improve the antibacterial properties, but also improve the survival rate, weight gain rate and crude protein content of Macrobrachium rosenbergii. β-alanine methyl ester derivatives and isomalt can act on a variety of pathogens that may exist in the aquaculture water and reduce the incidence of diseases; β-alanine methyl ester derivatives contain amino and imino structures, rich in N elements, and also contain β-alanine groups. Combined with the polyhydroxy structure in isomalt, it may be able to promote the expression of related protein genes in Macrobrachium rosenbergii cells, thereby increasing the protein content of Macrobrachium rosenbergii.

[0029] 3. In the seawater formula for Macrobrachium rosenbergii seedlings, the combination of 3-mercapto-2-butanone, β-alanine methyl ester derivatives and isomalt can further improve the antibacterial properties of the obtained artificial seawater, and can further improve the survival rate, weight gain rate and crude protein content of Macrobrachium rosenbergii. This may be because the guanidine group in the β-alanine methyl ester derivative and the hydroxyl group in isomalt and the thiol group and carbonyl group in 3-mercapto-2-butanone can further enhance the effect on pathogens and improve the antibacterial properties. DETAILED DESCRIPTION

[0030] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0031] Example 1 Seawater formula for Macrobrachium rosenbergii seedling rearing In the seawater formula, the mass proportions of sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA decrease in order, among which sodium chloride accounts for 69.32 parts, magnesium sulfate accounts for 26.34 parts, anhydrous calcium chloride accounts for 2.08 parts, potassium chloride accounts for 1.25 parts, potassium bromide accounts for 0.14 parts, boric acid accounts for 0.83 parts, and EDTA accounts for 0.03 parts.

[0032] Preparation method of seawater formula for Macrobrachium rosenbergii seedling cultivation The group mixing method is adopted to mix the groups during the feeding process, including the following steps: 1) Mix sodium chloride, potassium bromide and EDTA to prepare mother salt.

[0033] 2) Mix magnesium sulfate, calcium chloride, potassium chloride, boric acid and the prepared mother salt.

[0034] 3) Step 2) Add sodium chloride again to the mixed salt obtained, mix thoroughly, and obtain a seawater formula.

[0035] In the obtained seawater formula, the mass proportions of sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA decrease in order, among which sodium chloride accounts for 69.32 parts, magnesium sulfate accounts for 26.34 parts, anhydrous calcium chloride accounts for 2.08 parts, potassium chloride accounts for 1.25 parts, potassium bromide accounts for 0.14 parts, boric acid accounts for 0.83 parts, and EDTA accounts for 0.03 parts.

[0036] The mass ratio of sodium chloride used in step 1) to that used in step 3) is 1:100.

[0037] Method for using the seawater formula for Macrobrachium rosenbergii seedling rearing When using the seawater formula for Macrobrachium rosenbergii seedling rearing, add 14.5 kg of the seawater formula for every cubic meter of target seawater.

[0038] As needed, when using the seawater formula, additional anhydrous calcium chloride can be added. The amount of additional anhydrous calcium chloride added is 0.03 kg per cubic meter of water.

[0039] Example 2 The difference between this embodiment and embodiment 1 is that: when preparing the seawater formula, after step 3), β-alanine methyl ester derivative and isomalt are further added, the mass ratio of β-alanine methyl ester derivative and isomalt is 1:1, and the mass ratio of β-alanine methyl ester derivative to sodium chloride in the seawater formula is 1:100. Other conditions and steps are the same.

[0040] The β-alanine methyl ester derivative contains a β-alanine ester group and a guanidine structure. Specifically, the preparation method of the β-alanine methyl ester derivative is as follows: S1. Under ice bath conditions, add benzoyl isothiocyanate to a DCM solution containing β-alanine methyl ester hydrochloride, stir at 25°C, 150 r / min for 24 h, then filter, wash the filter residue with DCM, and dry it in vacuo at 55°C for 6 h.

[0041] The molar ratio of benzoyl isothiocyanate to β-alanine methyl ester hydrochloride is 1.2: 1. The mass volume ratio of β-alanine methyl ester hydrochloride to DCM is 1 g: 80 mL. When the filter residue obtained by filtration is washed with DCM, the mass volume ratio of the filter residue to DCM is 1 g: 50 mL, and the mixture is washed 3 times.

[0042] S2. The solid obtained in S1 was dissolved in ethanol, and NaOH solution was added dropwise thereto. The mixture was stirred at 25°C and 150 r / min for 24 h, and then filtered. The obtained filter residue was washed with ethanol and dried in vacuum at 55°C for 6 h.

[0043] Among them, the mass volume ratio of the solid obtained from S1 to ethanol is 1g:20mL; the concentration of the NaOH solution is 5.0mol / L, and the mass volume ratio of the solid obtained from S1 to the NaOH solution is 1g:2mL.

[0044] S3. The solid obtained in S2 was dissolved in ethanol and CH 3 I, 40°C, 150r / min stirring reaction for 48h, then filtered, the obtained filter residue was washed with ethanol and vacuum dried at 55°C for 6h.

[0045] The mass volume ratio of the solid obtained from S2 to ethanol is 1g:50mL; the mass volume ratio of the solid obtained from S2 to CH 3 The mass ratio of I is 1:2.

[0046] S4. The solid obtained in S3, n-propylamine and N,N-diisopropylethylamine were added to anhydrous DMF, and the mixture was stirred at 60°C and 150 r / min for 72 h. Then, a NaCl solution was added thereto, and the mixture was dialyzed with deionized water to obtain a β-alanine methyl ester derivative.

[0047] Among them, the mass volume ratio of the solid obtained from S3 and n-propylamine is 1 mg:1 mL; the volume ratio of n-propylamine to N,N-diisopropylethylamine is 1:0.2; the mass volume ratio of the solid obtained from S3 to DMF is 15 mg:1 mL.

[0048] The mass concentration of the NaCl solution was 20%, and the mass volume ratio of the solid obtained from S3 to the NaCl solution was 1 mg:5 mL; the MWCO of the membrane used for dialysis was 1000 Da.

[0049] Example 3 The difference between this embodiment and embodiment 1 is that: when preparing the seawater formula, after step 3), β-alanine methyl ester derivative and isomalt are further added, the mass ratio of β-alanine methyl ester derivative and isomalt is 1:1, and the mass ratio of β-alanine methyl ester derivative to sodium chloride in the seawater formula is 1:60. Other conditions and steps are the same.

[0050] Wherein, the preparation method of β-alanine methyl ester derivative is the same as that in Example 2.

[0051] Example 4 The difference between this embodiment and embodiment 1 is that: when preparing the seawater formula, after step 3), β-alanine methyl ester derivative and isomalt are further added, the mass ratio of β-alanine methyl ester derivative and isomalt is 1:1, and the mass ratio of β-alanine methyl ester derivative to sodium chloride in the seawater formula is 1:40. Other conditions and steps are the same.

[0052] Wherein, the preparation method of β-alanine methyl ester derivative is the same as that in Example 2.

[0053] Example 5 The difference between this embodiment and embodiment 1 is that: when preparing the seawater formula, after step 3), β-alanine methyl ester derivatives and isomalt are also added, the mass ratio of β-alanine methyl ester derivatives and isomalt is 1:1, and the mass ratio of β-alanine methyl ester derivatives to sodium chloride in the seawater formula is 1:100. In addition, 3-mercapto-2-butanone is additionally added, and the mass ratio of β-alanine methyl ester derivatives to 3-mercapto-2-butanone is 1:0.5. Other conditions and steps are the same.

[0054] Wherein, the preparation method of β-alanine methyl ester derivative is the same as that in Example 2.

[0055] Example 6 The difference between this embodiment and embodiment 1 is that: when preparing the seawater formula, after step 3), β-alanine methyl ester derivatives and isomalt are also added, the mass ratio of β-alanine methyl ester derivatives and isomalt is 1:1, and the mass ratio of β-alanine methyl ester derivatives to sodium chloride in the seawater formula is 1:100. In addition, 3-mercapto-2-butanone is additionally added, and the mass ratio of β-alanine methyl ester derivatives to 3-mercapto-2-butanone is 1:1. Other conditions and steps are the same.

[0056] Wherein, the preparation method of β-alanine methyl ester derivative is the same as that in Example 2.

[0057] Comparative Example 1 The difference between this comparative example and Example 1 is that: when preparing the seawater formula, after step 3), a β-alanine methyl ester derivative is further added, and the mass ratio of the β-alanine methyl ester derivative to the sodium chloride in the seawater formula is 1:50. Other conditions and steps are the same.

[0058] Wherein, the preparation method of β-alanine methyl ester derivative is the same as that in Example 2.

[0059] Comparative Example 2 The difference between this comparative example and Example 1 is that when preparing the seawater formula, isomalt is further added after step 3), and the mass ratio of isomalt to sodium chloride in the seawater formula is 1:50. Other conditions and steps are the same.

[0060] Test example The seawater formulas obtained in Examples 1-6 and Comparative Examples 1-2 were respectively used to prepare aquaculture water for aquaculture of Macrobrachium rosenbergii, and 14.5 kg of the seawater formula was added per cubic meter of target seawater. The Macrobrachium rosenbergii used had a body weight of 1.2±0.5 g, the water temperature during aquaculture was 25±2.5°C, oxygenation was maintained during aquaculture, 2 / 3 of the water was changed daily, and a compound feed without sulfonamide drugs was fed during aquaculture, 3 times a day, at 8:00, 14:00 and 20:00, respectively, and the daily feeding amount was 5%-20% of the mass of Macrobrachium rosenbergii, and the specific feeding amount needed to be adjusted according to the feeding situation of Macrobrachium rosenbergii.

[0061] The survival rates of Macrobrachium rosenbergii after 8 weeks of culture using the seawater formula obtained in each embodiment and comparative example are shown in Table 1.

[0062] Table 1 Survival rate of Macrobrachium rosenbergii in various embodiments and comparative examples

[0063] Referring to the data in Table 1, the survival rates of Macrobrachium rosenbergii in Examples 2-6 and Comparative Examples 1-2 were all improved relative to Example 1. Specifically, relative to Example 1, the survival rates of Macrobrachium rosenbergii in Examples 2-6 and Comparative Examples 1-2 were increased by 5.2%, 7.3%, 9.3%, 10.7%, 11.5%, 3.8%, and 2.8%, respectively. It can be seen that the addition of β-alanine methyl ester derivatives and isomalt to the seawater formula used for breeding Macrobrachium rosenbergii can greatly improve the survival rate of Macrobrachium rosenbergii.

[0064] Combined with the data of Examples 2-4, it can be seen that within a certain range, the content of β-alanine methyl ester derivatives and isomalt in the seawater formula is positively correlated with the survival rate of Macrobrachium rosenbergii.

[0065] Compared with Example 2, the survival rates of Macrobrachium rosenbergii in Examples 5-6 were increased by 5.2% and 6.0%, respectively. It can be seen that the additional addition of 3-mercapto-2-butanone to the seawater formula can further improve the survival rate of Macrobrachium rosenbergii.

[0066] Compared with Example 2, the survival rates of Macrobrachium rosenbergii in Comparative Examples 1-2 were reduced by 1.3% and 2.2%, respectively. It can be seen that in the seawater formula, in addition to the basic inorganic salts, the combined addition of β-alanine methyl ester derivatives and isomalt has a better effect on the survival rate of Macrobrachium rosenbergii than the addition of β-alanine methyl ester derivatives alone and isomalt alone.

[0067] After culturing Macrobrachium rosenbergii for 8 weeks using the seawater formula obtained in each embodiment and comparative example, Macrobrachium rosenbergii was fasted for 24 hours and the final body weight was measured. The weight gain rate is shown in Table 2.

[0068] Table 2 Weight gain rate of Macrobrachium rosenbergii in each embodiment and comparative example

[0069] Referring to the data in Table 2, the weight gain rates of Macrobrachium rosenbergii in Examples 2-6 and Comparative Examples 1-2 were all improved relative to Example 1. Specifically, relative to Example 1, the weight gain rates of Macrobrachium rosenbergii in Examples 2-6 and Comparative Examples 1-2 were increased by 8.0%, 9.4%, 10.8%, 12.0%, 12.9%, 4.5%, and 3.8%, respectively. It can be seen that adding β-alanine methyl ester derivatives and isomalt to the seawater formula used for breeding Macrobrachium rosenbergii can greatly improve the weight gain rate of Macrobrachium rosenbergii.

[0070] Combined with the data of Examples 2-4, it can be seen that within a certain range, the content of β-alanine methyl ester derivatives and isomalt in the seawater formula is positively correlated with the weight gain rate of Macrobrachium rosenbergii.

[0071] Compared with Example 2, the weight gain rates of Macrobrachium rosenbergii in Examples 5-6 were increased by 3.7% and 4.6%, respectively. It can be seen that the additional addition of 3-mercapto-2-butanone to the seawater formula can further increase the weight gain rate of Macrobrachium rosenbergii.

[0072] Compared with Example 2, the weight gain rates of Macrobrachium rosenbergii in Comparative Examples 1-2 were reduced by 3.5% and 3.8%, respectively. It can be seen that in the seawater formula, in addition to the basic inorganic salts, the combined addition of β-alanine methyl ester derivatives and isomalt has a better effect on the weight gain rate of Macrobrachium rosenbergii than the addition of β-alanine methyl ester derivatives alone and isomalt alone.

[0073] After culturing Macrobrachium rosenbergii for 8 weeks using the seawater formula obtained in each embodiment and comparative example, the crude protein content of Macrobrachium rosenbergii was measured by Kjeldahl nitrogen determination method, see Table 3.

[0074] Table 3 Crude protein content of Macrobrachium rosenbergii in each embodiment and comparative example

[0075] Referring to the data in Table 3, the crude protein content of Macrobrachium rosenbergii in Examples 2-6 and Comparative Examples 1-2 is improved relative to that in Example 1. Specifically, relative to Example 1, the crude protein content of Macrobrachium rosenbergii in Examples 2-6 and Comparative Examples 1-2 is increased by 6.2%, 8.7%, 11.4%, 12.6%, 13.9%, 4.1%, and 3.0%, respectively. It can be seen that the addition of β-alanine methyl ester derivatives and isomalt to the seawater formula used for breeding Macrobrachium rosenbergii can greatly increase the crude protein content of Macrobrachium rosenbergii.

[0076] Combined with the data of Examples 2-4, it can be seen that within a certain range, the contents of β-alanine methyl ester derivatives and isomalt in the seawater formula are positively correlated with the crude protein content of Macrobrachium rosenbergii.

[0077] Compared with Example 2, the crude protein content of Macrobrachium rosenbergii in Examples 5-6 was increased by 6.1% and 7.3%, respectively. It can be seen that the additional addition of 3-mercapto-2-butanone to the seawater formula can further increase the crude protein content of Macrobrachium rosenbergii.

[0078] Compared with Example 2, the crude protein content of Macrobrachium rosenbergii in Comparative Examples 1-2 was reduced by 1.9% and 2.9%, respectively. It can be seen that in the seawater formula, in addition to the basic inorganic salts, the combined addition of β-alanine methyl ester derivatives and isomalt has a better effect on the crude protein content of Macrobrachium rosenbergii than the addition of β-alanine methyl ester derivatives alone and isomalt alone.

[0079] The conventional operations in the operating steps of the present invention are well known to those skilled in the art and will not be described in detail here.

[0080] The above-described embodiments provide a detailed description of the technical solution of the present invention. It should be understood that the above-described embodiments are only specific embodiments of the present invention and are not intended to limit the present invention. Any person skilled in the art who is familiar with the present technical field can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims

1. A seawater formula for breeding Macrobrachium rosenbergii seedlings, comprising sodium chloride, magnesium sulfate, anhydrous calcium chloride, potassium chloride, potassium bromide, boric acid and EDTA; and also comprising a β-alanine methyl ester derivative and isomalt; the β-alanine methyl ester derivative contains a guanidine structure.

2. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 1, characterized in that: Sodium chloride accounts for 65-75 parts, magnesium sulfate accounts for 20-28 parts, anhydrous calcium chloride accounts for 1-3 parts, potassium chloride accounts for 0.5-2 parts, potassium bromide accounts for 0.1-0.3 parts, boric acid accounts for 0.5-1.5 parts, and EDTA accounts for 0.01-0.05 parts.

3. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 1, characterized in that: The mass ratio of the beta-alanine methyl ester derivative to isomalt is 1:0.5-1.5, and the mass ratio of the beta-alanine methyl ester derivative to the sodium chloride in the seawater formula is 1:30-120.

4. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 1, characterized in that: When used, 13-16 kg of the seawater formula is added per cubic meter of target seawater.

5. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 1, characterized in that: The preparation method of the β-alanine methyl ester derivative comprises the following steps: S1. Add benzoyl isothiocyanate to a DCM solution containing β-alanine methyl ester hydrochloride, stir the reaction, and then filter. The resulting filter residue is washed and dried. S2. The solid obtained in S1 was reacted with a NaOH solution, and then filtered, and the resulting residue was washed and dried; S3. The solid obtained from S2 was reacted with CH3I and then filtered, and the resulting residue was washed and dried; S4. The solid obtained in S3, n-propylamine and N,N-diisopropylethylamine were added to anhydrous DMF and stirred for reaction; then, a NaCl solution was added thereto and dialyzed with deionized water to obtain a β-alanine methyl ester derivative.

6. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 5, characterized in that: In S1, the molar ratio of benzoyl isothiocyanate to β-alanine methyl ester hydrochloride is 1-1.5:1; In S1, the mass volume ratio of β-alanine methyl ester hydrochloride to DCM is 1 g:60-120 mL.

7. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 5, characterized in that: In S2, the solid obtained in S1 is dissolved in ethanol and then reacted with NaOH solution, and the mass volume ratio of NaOH to ethanol is 1g:15-40mL; In S2, the concentration of the NaOH solution is 3.0-6.0 mol / L; the mass volume ratio of the solid obtained in S1 to the NaOH solution is 1 g:1-5 mL.

8. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 5, characterized in that: In S3, the solid obtained from S2 is dissolved in ethanol and then reacted with CH3I; the mass volume ratio of the solid obtained from S2 to ethanol is 1g:30-80mL; In S3, the mass ratio of the solid obtained in S2 to CH3I is 1:1-3.

9. The seawater formula for breeding Macrobrachium rosenbergii seedlings according to claim 5, characterized in that: In S4, the mass concentration of the NaCl solution is 15-40%; the mass volume ratio of the solid obtained in S3 to the NaCl solution is 1 mg:3-8 mL.

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