A disinfectant for red claw crayfish with eggs, and its preparation method and application

Abstract

The invention discloses a disinfectant for red claw crayfish with eggs, and a preparation method and application thereof. The solvent of the invention is eugenol, and the solute includes acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E; the concentrations of the solutes are respectively: acacia essential oil 10-12%, cinnamaldehyde 18-25g / L, trehalose 0.12-0.15mol / L, vitamin C 7-10g / L, vitamin A 2.5-6.0g / L, and vitamin E 2.5-6.0g / L. The invention also provides a preparation method for the above disinfectant. In the invention, the solute and the solvent complement each other, cooperate with each other, promote each other, have good disinfection effect, avoid infection of shrimp eggs, are non-toxic and harmless, have a small amount of use, and also have an anesthetic function, reduce the loss of shrimp eggs caused by stress spring tail due to stimulation during the grasping process of the egg-bearing shrimp, protect the parent shrimp and embryos, are easy to use, and have a high emergence rate.

Description

Technical Field

[0001] The invention belongs to the technical field of red claw crayfish breeding, and particularly relates to a disinfectant for red claw crayfish carrying eggs, and a preparation method and application thereof. Background Art

[0002] Red claw crayfish (Cherax quadricarinatus) is an animal of the family Pseudocranidae and the genus Cherax. It has the advantages of fast growth, high meat content, beautiful appearance, and low incidence of diseases. It is also resistant to low oxygen, has tender meat, and tastes delicious, and is recognized by the market. The main breeding method of red claw crayfish is indoor cement pool breeding. The breeding process is roughly as follows: the parent shrimp enters the greenhouse in October every year, and is cultivated over the winter until the following year. After mating, the water in the cement pool is drained, and the shrimps with eggs are caught and put into the floating box, and cultivated until the shrimps are empty; the empty shrimps and female shrimps without eggs are re-mated with male shrimps in the new cement pool, and the shrimps with eggs are obtained again, and are caught and cultivated in the floating box; under normal circumstances, a female shrimp can mate and lay eggs more than three times during the nursery cycle.

[0003] In the breeding process of red claw crayfish, the parent shrimps need to be cultivated in cement ponds for a long time before the brooding shrimps can be obtained. During the cultivation of these brooding shrimps in the floating tanks, the eggs are prone to infection, mold or death during the hatching period. Dead eggs, moldy eggs and infected eggs may fall off from the mother, resulting in a low hatching rate of shrimp fry and insufficient fry output, which hinders the development of the industry.

[0004] In the prior art, the disinfectants commonly used in the breeding process of freshwater crayfish include formaldehyde, hydrogen peroxide, potassium permanganate, copper sulfate, povidone iodine or isopropyl alcohol, among which formaldehyde is the most effective disinfectant, and other chemical reagents such as hydrogen peroxide, potassium permanganate, copper sulfate, povidone iodine or isopropyl alcohol have poor disinfection effects; however, formaldehyde is harmful to the environment and human body, and the use of formaldehyde has been restricted in many places. Therefore, it is urgent to develop an effective disinfectant widely used in the breeding process of red claw crayfish. Summary of the invention

[0005] The purpose of the present invention is to provide a disinfectant for red claw crayfish and its preparation method and application, aiming to solve the problems in the prior art that formaldehyde disinfectant used in the breeding process of freshwater crayfish is harmful to the environment and human body, thereby limiting its application and other disinfectants have poor disinfection effects.

[0006] In order to solve the above technical problems, the present invention is mainly implemented through the following technical solutions:

[0007] In one aspect, the present invention discloses a disinfectant for red claw crayfish with eggs, wherein the solvent of the disinfectant is eugenol, and the solute of the disinfectant comprises acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E; the concentrations of the solutes in the disinfectant are: acacia essential oil 10-12%, cinnamaldehyde 18-25 g / L, trehalose 0.12-0.15 mol / L, vitamin C 7-10 g / L, vitamin A 2.5-6.0 g / L, and vitamin E 2.5-6.0 g / L.

[0008] The disinfectant for red claw crayfish with eggs of the present invention uses eugenol as solvent. Eugenol is a pure natural substance with low toxicity, strong solubility, disinfection effect, strong antibacterial effect on various fungi, bacteria and molds, and anesthetic effect. It has no residual period and is a legal aquatic anesthetic. Acacia essential oil is extracted from plants, is non-toxic and harmless, and is a green ecological disinfectant. Cinnamaldehyde is an aldehyde organic compound, a yellow viscous liquid, which exists in large quantities in plants such as cinnamon. It has bactericidal, disinfectant and antiseptic effects, especially for fungi. It has a certain hydrophilicity and is easily adsorbed on the hydrophilic group on the surface of fungi. It has a certain inhibitory effect on the activity of chitin synthase and glucan synthase on the cell membrane, thereby inhibiting the production of chitin and glucose in the cell wall, destroying the polysaccharide structure of the cell wall, and inhibiting the growth of bacteria. It can also dissolve between adjacent fatty acyl chains in the hydrophobic domain of the cell membrane, enhancing the permeability of the cell membrane, leading to ATP leakage and bacterial death. At the same time, it has an antioxidant effect and can alleviate the stress of shrimp and embryos during anesthesia. The invention compounds eugenol, acacia essential oil and cinnamaldehyde, wherein the phenolic hydroxyl group in the eugenol is combined with the aldehyde group in the cinnamaldehyde by a strong van der Waals force under the action of the carbon-carbon double bond, and the acacia essential oil has multiple functional groups such as a hydroxyl group and an aldehyde group, and the acacia essential oil is strongly combined with the eugenol and the cinnamaldehyde, and the three promote each other, synergistically enhance the effect, and have a good disinfection effect, thereby overcoming the disadvantages of high cost, fast volatilization, poor stability and slow effect of single eugenol as a disinfectant, avoiding the problem that the single acacia essential oil as a disinfectant is not suitable for actual production application due to too short disinfection time, and also overcoming the disadvantages of poor stability, easy volatility, easy oxidation and poor sterilization effect of single cinnamaldehyde as a disinfectant; and at the same time, eugenol and cinnamaldehyde also have an anesthetic effect, thereby reducing the problem that the red claw crayfish with eggs may lose shrimp eggs due to stress tail springing due to stimulation during the catching process. The invention also adds trehalose, vitamin C, vitamin A and vitamin E, improves the stability of eugenol, acacia essential oil and cinnamaldehyde, maintains the cell membrane stability and permeability of the embryo in the disinfectant, protects the embryo, prevents the embryo from being damaged, and further improves the germination rate.

[0009] As a preferred embodiment, the concentrations of the solutes in the disinfectant are: acacia essential oil 10-12%, cinnamaldehyde 20-25g / L, trehalose 0.12-0.15mol / L, vitamin C 7-9g / L, vitamin A 3.0-5.0g / L, vitamin E 3.0-5.0g / L. In the present invention, trehalose is a typical stress metabolite, which can form a unique protective film on the cell surface under harsh environmental conditions such as high temperature, high cold, high osmotic pressure and dehydration, effectively protecting the biological molecular structure from being destroyed, thereby maintaining the life process and biological characteristics of the living body; when the embryo is under the action of eugenol, acacia essential oil and cinnamaldehyde, the stability and permeability of the cell membrane are at risk of change, trehalose can effectively form a unique protective film on the cell membrane surface to prevent the embryo from being damaged. Vitamin C, vitamin A and vitamin E are all antioxidants that can reduce the stress of shrimp parents and shrimp eggs, neutralize harmful free radicals, and help improve the stability of eugenol, acacia essential oil and cinnamaldehyde.

[0010] As a preferred embodiment, the concentrations of the solutes in the disinfectant are: 11% acacia essential oil, 21.5g / L cinnamaldehyde, 0.13mol / L trehalose, 8g / L vitamin C, 4.5g / L vitamin A, and 4.5g / L vitamin E. The disinfectant of the present invention is a composite disinfectant and anesthesia reagent composed of eugenol, acacia essential oil and cinnamaldehyde, and trehalose, vitamin C, vitamin A and vitamin E are added to form a disinfectant for red claw crayfish brooding shrimp, which is used in the breeding process of red claw crayfish, specifically, in the maternal hatching process of red claw crayfish, to reduce the stress of parent shrimp and embryos, protect embryos and increase the stability of the reagent, and the solvent and solute are scientifically compounded, mutually coordinated, and the use effect is good. In the disinfectant of the present invention, the optimal ratio of eugenol, acacia essential oil and cinnamaldehyde is preferably 40mL:5mL:1g, and the concentration of acacia essential oil is volume percentage.

[0011] In another aspect, a method for preparing a disinfectant for red claw crayfish comprising the following steps: 1) weighing eugenol, acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E in sequence; 2) adding acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, stirring at 30-35° C. for 5-10 minutes to obtain a disinfectant.

[0012] The preparation method of the disinfectant for red claw crayfish egg-carrying shrimps of the present invention is simple to operate, convenient to prepare, mild in conditions, has no special requirements on equipment, is easy to realize industrialization, is widely used in the process of hatching red claw crayfish mothers, and meets the needs of the large-scale breeding process of red claw crayfish.

[0013] As a preferred embodiment, in the step 2), the stirring speed is 30-60 r / min. Stirring promotes the dissolution of acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E in eugenol. Appropriate stirring accelerates the dissolution process, increases the dissolution rate and is easy to control.

[0014] As a preferred embodiment, in step 2), the stirring is clockwise stirring. The stirring is usually carried out in one direction, for example, clockwise stirring, so that the acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E are quickly dissolved in eugenol; such stirring in the same direction not only has a good stirring effect, but also does not cause material overflow, and is easy to operate.

[0015] As a preferred embodiment, in the step 2), the temperature is heated to 30-35° C. by water bath. The present invention controls the dissolution temperature of acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E in eugenol, increases the dissolution rate, and promotes the rapid interaction between eugenol, acacia essential oil and cinnamaldehyde, thereby improving the comprehensive performance of the disinfectant.

[0016] In another aspect, the present invention provides an application of a disinfectant for red claw crayfish brooding shrimps, wherein the disinfectant for red claw crayfish brooding shrimps is placed in a cement pool of red claw crayfish brooding shrimps before catching the red claw crayfish brooding shrimps. The disinfectant of the present invention is used before catching the red claw crayfish brooding shrimps, and the red claw crayfish brooding shrimps are soaked in water containing the disinfectant to be fully disinfected. The disinfectant is easy to use, easy to operate, and has a good use effect. Normally, when the disinfectant for red claw crayfish brooding shrimps of the present invention is used, the water temperature is 28-30°C, which is just the appropriate temperature for embryonic development.

[0017] As a preferred embodiment, the amount of the disinfectant for red claw crayfish in the cement pool of red claw crayfish is 0.5-1.0‰ of the water mass. The disinfectant of the present invention is directly put into the cement pool of red claw crayfish in the cement pool, and 0.5-1.0L of the disinfectant is added to every 1000L of water in the cement pool. The amount of the disinfectant is small and the disinfection effect is good.

[0018] As a preferred embodiment, the disinfection time of the disinfectant for red claw crayfish in the cement pool of red claw crayfish is 5-8 minutes. After the disinfectant of the present invention is evenly mixed into the water in the cement pool, after 5-8 minutes, the red claw crayfish in the cement pool can be fully disinfected and anesthetized, and at this time, the red claw crayfish can be easily caught.

[0019] Compared with the prior art, the invention has the following beneficial effects: the eugenol, acacia essential oil and cinnamaldehyde in the disinfectant of the invention form a composite disinfectant, the three interact with each other, promote each other, synergize and enhance the effect, have good disinfection effect, avoid infection of shrimp eggs, are non-toxic and harmless, and have low cost; at the same time, the eugenol and cinnamaldehyde also have an anesthetic function, which gives the disinfectant an anesthetic effect, and reduces the problem of shrimp egg loss caused by tail stress due to stimulation of red claw crayfish carrying eggs during the catching process; trehalose, vitamin C, vitamin A and vitamin E are also added to the disinfectant of the invention, and the trehalose, vitamin C, vitamin A and vitamin E are organically compounded, which improves the stability of eugenol, acacia essential oil and cinnamaldehyde, maintains the cell membrane stability and permeability of embryos in the disinfectant, protects the embryos, prevents the embryos from being damaged, is easy to use, has a suitable disinfection time, improves the emergence rate, is widely used in the red claw crayfish mother hatching process, and meets the needs of the red claw crayfish large-scale breeding process. DETAILED DESCRIPTION

[0020] The technical solution of the present invention will be clearly and completely described below in conjunction with the specific embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] The invention discloses a disinfectant for red claw crayfish with eggs. The solvent of the disinfectant is eugenol, and the solute of the disinfectant comprises acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E. The concentrations of the solutes in the disinfectant are respectively: 10-12% of acacia essential oil, 18-25 g / L of cinnamaldehyde, 0.12-0.15 mol / L of trehalose, 7-10 g / L of vitamin C, 2.5-6.0 g / L of vitamin A and 2.5-6.0 g / L of vitamin E.

[0022] Preferably, the concentrations of the solutes in the disinfectant are: acacia essential oil 10-12%, cinnamaldehyde 20-25 g / L, trehalose 0.12-0.15 mol / L, vitamin C 7-9 g / L, vitamin A 3.0-5.0 g / L, and vitamin E 3.0-5.0 g / L.

[0023] Furthermore, the concentrations of solutes in the disinfectant are: acacia essential oil 11%, cinnamaldehyde 21.5 g / L, trehalose 0.13 mol / L, vitamin C 8 g / L, vitamin A 4.5 g / L, and vitamin E 4.5 g / L.

[0024] The method for preparing a disinfectant for red claw crayfish with eggs of the present invention comprises the following steps:

[0025] 1) Weigh eugenol, acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E in sequence;

[0026] 2) Add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, stir at 30-35° C. for 5-10 minutes to obtain a disinfectant.

[0027] Preferably, in step 2), the stirring speed is 30-60 r / min.

[0028] Preferably, in step 2), the stirring is performed in a clockwise direction.

[0029] Preferably, in step 2), the mixture is heated in a water bath to a temperature of 30-35°C.

[0030] The invention discloses an application of a disinfectant for red claw crayfish carrying eggs. The disinfectant for red claw crayfish carrying eggs is put into a cement pool of the red claw crayfish carrying eggs before the red claw crayfish carrying eggs are caught.

[0031] Preferably, the amount of the disinfectant for red claw crayfish in the cement pool containing eggs is 0.5-1.0‰ of the water mass.

[0032] Preferably, the disinfection time of the disinfectant for red claw crayfish in the cement pool of red claw crayfish is 5-8 minutes.

[0033] Embodiment 1

[0034] The method for preparing a disinfectant for red claw crayfish with eggs of the present invention comprises the following steps:

[0035] 1) Take 580 mL of eugenol, 81.6 mL of acacia essential oil, 17 g of cinnamaldehyde, 102 mmol of trehalose, 4.76 g of vitamin C, 4.08 g of vitamin A and 4.08 g of vitamin E in sequence;

[0036] 2) Heat eugenol to 30° C. in a water bath, add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, and stir for 10 minutes to obtain a disinfectant.

[0037] The concentrations of the solutes in the disinfectant reagent are: 12% acacia essential oil, 25 g / L cinnamaldehyde, 0.15 mol / L trehalose, 7 g / L vitamin C, 6.0 g / L vitamin A and 6.0 g / L vitamin E.

[0038] Embodiment 2

[0039] The method for preparing a disinfectant for red claw crayfish with eggs of the present invention comprises the following steps:

[0040] 1) Take 620 mL of eugenol, 70 mL of acacia essential oil, 12.6 g of cinnamaldehyde, 84 mmol of trehalose, 7 g of vitamin C, 1.75 g of vitamin A and 1.75 g of vitamin E in sequence;

[0041] 2) Heat eugenol to 35° C. in a water bath, add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, stir clockwise at a speed of 30 r / min for 5 min, and obtain a disinfectant reagent.

[0042] The concentrations of the solutes in the disinfectant reagent are: 10% acacia essential oil, 18 g / L cinnamaldehyde, 0.12 mol / L trehalose, 10 g / L vitamin C, 2.5 g / L vitamin A and 2.5 g / L vitamin E.

[0043] Embodiment 3

[0044] The method for preparing a disinfectant for red claw crayfish with eggs of the present invention comprises the following steps:

[0045] 1) Take 600 mL of eugenol, 74.8 mL of acacia essential oil, 13.6 g of cinnamaldehyde, 88.4 mmol of trehalose, 6.12 g of vitamin C, 2.04 g of vitamin A and 3.40 g of vitamin E in sequence;

[0046] 2) Heat eugenol to 33° C. in a water bath, add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, stir clockwise at a speed of 60 r / min for 7 min, and obtain a disinfectant reagent.

[0047] The concentrations of the solutes in the disinfectant reagent are: 11% of acacia essential oil, 20 g / L of cinnamaldehyde, 0.13 mol / L of trehalose, 9 g / L of vitamin C, 3.0 g / L of vitamin A, and 5.0 g / L of vitamin E.

[0048] Embodiment 4

[0049] The method for preparing a disinfectant for red claw crayfish with eggs of the present invention comprises the following steps:

[0050] 1) Take 600 mL of eugenol, 84 mL of acacia essential oil, 16.8 g of cinnamaldehyde, 98 mmol of trehalose, 5.6 g of vitamin C, 3.5 g of vitamin A and 2.1 g of vitamin E in sequence;

[0051] 2) Heat eugenol to 34° C. in a water bath, add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, and stir for 9 minutes to obtain a disinfectant.

[0052] The concentrations of solutes in the disinfectant reagent are: 12% acacia essential oil, 24 g / L cinnamaldehyde, 0.14 mol / L trehalose, 8 g / L vitamin C, 5.0 g / L vitamin A, and 3.0 g / L vitamin E.

[0053] Embodiment 5

[0054] The method for preparing a disinfectant for red claw crayfish with eggs of the present invention comprises the following steps:

[0055] 1) Take 600 mL of eugenol, 74.8 mL of acacia essential oil, 14.62 g of cinnamaldehyde, 88.4 mmol of trehalose, 5.44 g of vitamin C, 3.06 g of vitamin A and 3.06 g of vitamin E in sequence;

[0056] 2) Heat eugenol to 34° C. in a water bath, add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, stir clockwise at a speed of 50 r / min for 9 minutes, and obtain a disinfectant reagent.

[0057] The concentrations of solutes in the disinfectant reagent are: 11% of acacia essential oil, 21.5 g / L of cinnamaldehyde, 0.13 mol / L of trehalose, 8 g / L of vitamin C, 4.5 g / L of vitamin A, and 4.5 g / L of vitamin E.

[0058] Comparative Example 1

[0059] Take 600 mL of eugenol as a disinfectant and record it as control sample 1.

[0060] Comparative Example 2

[0061] Take 600 mL of eugenol and 74 mL of acacia essential oil, heat the eugenol to 34° C. in a water bath, add the acacia essential oil to the eugenol, and stir clockwise at a speed of 50 r / min for 9 min as a disinfectant, wherein the volume concentration of the acacia essential oil is 11%, recorded as control sample 2.

[0062] Comparative Example 3

[0063] Take 600 mL of eugenol, 74.8 mL of acacia essential oil and 14.62 g of cinnamaldehyde, heat the eugenol to 34°C in a water bath, add acacia essential oil and cinnamaldehyde to the eugenol, and stir clockwise at a speed of 50 r / min for 9 minutes as a disinfectant, wherein the volume concentration of acacia essential oil is 11%, and the mass concentration of cinnamaldehyde is 21.5 g / L, which is recorded as control sample three.

[0064] Comparative Example 4

[0065] Take 600 mL of eugenol, 64.6 mL of acacia essential oil, 20.4 g of cinnamaldehyde, 122 mmol of trehalose, 4 g of vitamin C, 4.8 g of vitamin A and 5.0 g of vitamin E, heat eugenol to 34 ° C in a water bath, add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, and stir clockwise at a speed of 50 r / min for 9 min as a disinfectant, wherein the volume concentration of acacia essential oil is 9.5%, the mass concentration of cinnamaldehyde is 30 g / L, trehalose is 0.18 mol / L, vitamin C is 5.9 g / L, vitamin A is 7.1 g / L, and vitamin E is 7.4 g / L, which is recorded as control sample four.

[0066] Comparative Example 5

[0067] Take 600 mL of eugenol, 88 mL of acacia essential oil, 11 g of cinnamaldehyde, 68 mmol of trehalose, 7.5 g of vitamin C, 1.4 g of vitamin A and 1.2 g of vitamin E, heat eugenol to 34 ° C in a water bath, add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, and stir clockwise at a speed of 50 r / min for 9 min as a disinfectant, wherein the volume concentration of acacia essential oil is 12.9%, the mass concentration of cinnamaldehyde is 16.2 g / L, trehalose is 0.10 mol / L, vitamin C is 11.0 g / L, vitamin A is 2.1 g / L, and vitamin E is 1.8 g / L, which is recorded as control sample five.

[0068] Experiment 1

[0069] The five disinfectants obtained from Examples 1 to 5 of the present invention and the five disinfectants obtained from Comparative Examples 1 to 5 were respectively subjected to disinfection application experiments. Among them, the red claw crayfish used in the experiment came from the Zhejiang Freshwater Fisheries Research Institute. On October 15, 2021, the healthy broodstock with complete limbs and basically the same specifications were introduced into the greenhouse and evenly dispersed into 10 cement pools, with 30 broodstock / m in each cement pool. 2, uniformly cultivate overwintering and mate; on March 20, 2022, when the embryos of the egg-carrying shrimps are cultivated to the stage of seven to eighteen pairs of appendages and are red or orange, part of the water in the cement pool is drained to make the remaining water volume in the cement pool be 1500L, and the water temperature is adjusted to 28°C, and five parts of disinfectants obtained in Examples 1 to 5 of the present invention and five parts of disinfectants obtained in Comparative Examples 1 to 5 are added to the above 10 cement pools, respectively, and the added amount is 0.5‰ of the water mass in the cement pool. The shrimps are evenly dispersed and disinfected for 5 minutes, and the water in the cement pool is drained again; in order to facilitate experimental statistics, the in vitro hatching method is adopted, the egg-carrying shrimps are grabbed into the floating box, resuscitated, the eggs are peeled, and the in vitro hatching is carried out; the hatching and cultivation conditions of each group are the same, and the experiment is carried out until the seedlings emerge.

[0070] The recovery time and recovery rate during the experiment were statistically analyzed. The anesthesia time was defined as the time required for the shrimp to lie quietly at the bottom of the bucket, show no response to mechanical stimulation, and not flick its tail when grasped. After the shrimp was anesthetized, it was immediately transferred to fresh water of the same volume as the anesthesia water for still water recovery and continuous aeration. The time required for the shrimp behavior to fully return to normal was defined as the recovery time. The recovery rate was defined as the ratio of the number of shrimp that could be recovered after anesthesia to the total number of experimental shrimp. The experimental results are listed in Table 1.

[0071] The in vitro hatching technology is an existing mature technology. It usually uses circulating water and mechanically pulled embryo separation devices. The embryos are gently peeled off from the attached abdominal foot bristles by combs. The embryos peeled off from each egg-carrying shrimp are placed in an incubation box, and the incubation box (the upper width is 2.5 cm) is placed in the incubation device for incubation. No disinfection is performed after incubation, and the incubation water temperature is maintained at 28°C. After incubation, the number of 1st and 3rd stage juvenile shrimps is counted (the 1st stage juvenile shrimps refer to the juvenile shrimps that have just broken the membrane and hatched. After molting, they become the 2nd stage juvenile shrimps. After molting again, they become the 3rd stage juvenile shrimps. The 3rd stage juvenile shrimps can swim freely, marking the end of in vitro hatching), and the hatching rate (hatching rate % = the number of 1st stage juvenile shrimps / the number of embryos × 100%) and the emergence rate (emergence rate % = the number of 3rd stage juvenile shrimps / the number of embryos × 100%) are measured. During the incubation period, dead eggs and dead shrimps are regularly removed. All data are expressed as mean ± standard deviation, and statistical analysis was performed using Statistica software, and SPSS 17.0 computer program was used to perform variance analysis (ANOVA) on the results, and the data were subjected to arc sine transformation. For all statistical tests, P < 0.05 was considered significant, and the experimental results are also listed in Table 1.

[0072] It can be seen from Table 1 that the disinfection reagent for red claw crayfish with eggs obtained by the present invention has a certain anesthetic effect on red claw crayfish with eggs, and the anesthetized red claw crayfish with eggs can be completely revived, and the recovery rate is 100%; the recovery time of the red claw crayfish with eggs disinfected by the disinfection reagent for red claw crayfish with eggs of the present invention is between 135-172s, the recovery time is short, the recovery is fast, and the recovery time is sufficient to complete the capture of the red claw crayfish with eggs, thereby reducing the problem of shrimp egg loss caused by the stress tail flicking of the red claw crayfish with eggs during the capture process due to stimulation.

[0073] It can also be seen from Table 1 that the hatching rate of the shrimp eggs of the red claw crayfish brooding shrimp disinfected with the disinfectant for red claw crayfish obtained by the present invention is 87-95%, which is higher than the hatching rate of the shrimp eggs of the red claw crayfish brooding shrimp disinfected with the control sample 1, the control sample 2, the control sample 3, the control sample 4 and the control sample 5; and the shrimp egg germination rate of the red claw crayfish brooding shrimp disinfected with the disinfectant for red claw crayfish obtained by the present invention is 58.06-64.11%, which is also higher than the shrimp egg germination rate of the red claw crayfish brooding shrimp disinfected with the control sample 1, the control sample 2, the control sample 3, the control sample 4 and the control sample 5.

[0074] Table 1 Effects of not using disinfectants on the recovery, hatching rate and emergence rate of broodstock

[0075] Disinfectants Recovery rate (%) Recovery time(s) Hatching rate (%) Seedling rate (%) Embodiment 1 100 149-172 <![CDATA[89.07±3.23 d ]]> <![CDATA[58.06±2.20 c ]]> Embodiment 2 100 145-169 <![CDATA[89.24±1.76 d ]]> <![CDATA[59.35±2.31 c ]]> Embodiment 3 100 147-171 <![CDATA[88.99±2.14 d ]]> <![CDATA[59.59±2.86 c ]]> Embodiment 4 100 141-163 <![CDATA[91.82±4.86 d ]]> <![CDATA[60.71±3.88 c ]]> Embodiment 5 100 135-156 <![CDATA[94.65±1.45 d ]]> <![CDATA[64.11±2.31 c ]]> Control Sample 1 100 156-193 <![CDATA[65.74±8.66 a ]]> <![CDATA[43.44±1.95 a ]]> Control Sample 2 100 152-188 <![CDATA[73.56±3.78 b ]]> <![CDATA[49.02±2.58 b ]]> Control sample 3 100 154-186 <![CDATA[79.14±2.41 bc ]]> <![CDATA[50.62±2.35 b ]]> Control Sample 4 100 151-179 <![CDATA[81.94±1.91 c ]]> <![CDATA[51.73±3.79 b ]]> Control Sample 5 100 156-178 <![CDATA[81.85±2.87 c ]]> <![CDATA[51.70±2.22 b ]]>

[0076] Note: The same letters in the same column indicate no significant difference (P>0.05), and different letters in the same column indicate significant difference (P<0.05).

[0077] Therefore, compared with the prior art, the invention has the following beneficial effects: the eugenol, acacia essential oil and cinnamaldehyde in the disinfectant of the invention form a composite disinfectant, the three interact with each other, promote each other, synergize, have good disinfection effect, avoid infection of shrimp eggs, are non-toxic and harmless, and have low cost; at the same time, eugenol and cinnamaldehyde also have an anesthetic function, which gives the disinfectant an anesthetic effect, and reduces the problem of shrimp egg loss caused by tail stress caused by stimulation of red claw crayfish carrying eggs during the catching process; trehalose, vitamin C, vitamin A and vitamin E are also added to the disinfectant of the invention, and the trehalose, vitamin C, vitamin A and vitamin E are organically compounded, which improves the stability of eugenol, acacia essential oil and cinnamaldehyde, maintains the cell membrane stability and permeability of the embryo in the disinfectant, protects the embryo, prevents the embryo from being damaged, is easy to use, has a suitable disinfection time, improves the emergence rate, is widely used in the hatching process of red claw crayfish mothers, and meets the needs of the large-scale breeding process of red claw crayfish.

[0078] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A disinfectant for red claw crayfish, characterized in that: The solvent of the disinfectant is eugenol, and the solute of the disinfectant is composed of acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E; The concentrations of the solutes in the disinfectant reagent are respectively: 10-12% of acacia essential oil, 18-25 g / L of cinnamaldehyde, 0.12-0.15 mol / L of trehalose, 7-10 g / L of vitamin C, 2.5-6.0 g / L of vitamin A, and 2.5-6.0 g / L of vitamin E.

2. The disinfectant for red claw crayfish according to claim 1, characterized in that: The concentrations of the solutes in the disinfectant reagent are respectively: 10-12% of acacia essential oil, 20-25 g / L of cinnamaldehyde, 0.12-0.15 mol / L of trehalose, 7-9 g / L of vitamin C, 3.0-5.0 g / L of vitamin A, and 3.0-5.0 g / L of vitamin E.

3. The disinfectant for red claw crayfish according to claim 2, characterized in that: The concentrations of the solutes in the disinfectant reagent are: acacia essential oil 11%, cinnamaldehyde 21.5 g / L, trehalose 0.13 mol / L, vitamin C 8 g / L, vitamin A 4.5 g / L, and vitamin E 4.5 g / L.

4. A method for preparing a disinfectant for red claw crayfish according to any one of claims 1 to 3, characterized in that: The following steps are involved: 1) Weigh eugenol, acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E in sequence; 2) Add acacia essential oil, cinnamaldehyde, trehalose, vitamin C, vitamin A and vitamin E to eugenol in sequence, stir at 30-35° C. for 5-10 minutes to obtain a disinfectant.

5. The method for preparing a disinfectant for red claw crayfish according to claim 4, characterized in that: In the step 2), the stirring speed is 30-60 r / min.

6. The method for preparing the disinfectant for red claw crayfish according to claim 5, characterized in that: In the step 2), the stirring is performed in a clockwise direction.

7. The method for preparing a disinfectant for red claw crayfish according to claim 4, characterized in that: In the step 2), the mixture is heated in a water bath to a temperature of 30-35°C.

8. Use of the disinfectant for red claw crayfish with eggs according to any one of claims 1 to 4 in the preparation of a disinfectant.

9. Use of the disinfectant for red claw crayfish incubated with eggs according to claim 8 in the preparation of a disinfectant, characterized in that: The disinfectant for red claw crayfish with eggs is put into the cement pool of red claw crayfish with eggs before catching the red claw crayfish with eggs. The amount of the disinfectant for red claw crayfish with eggs added to the cement pool of red claw crayfish with eggs is 0.5-1.0‰ of the water mass.