阅读说明：本技术 一种复合工艺生产海藻水解产品及其在虾养殖中应用 (Seaweed hydrolysate produced by composite process and application thereof in shrimp culture ) 是由 周腾 于 2021-10-10 设计创作，主要内容包括：本发明涉及生物技术领域,具体涉及一种复合工艺生产海藻水解产品及其在虾养殖中应用；海藻水解产品由泡叶藻经切块、粉碎、水解以及过滤制备；采用本发明工艺生产出的海藻酸、甜菜碱含量分别为：≥45g/l、≥26g/l,本工艺未经强酸强碱高温处理、也未经高温浓缩处理,产品的活性物质能保持较高的活性,同时又达到较高的含量要求,从生产成本与生物活性都实现新的创新,海藻水解产品在对虾养殖过程中病害的预防和治疗中的应用,产品基本解决对虾养殖化学品滥用的问题,优于化学品防治的效果,特别是早期预防具有较好的效果。(The invention relates to the technical field of biology, in particular to a seaweed hydrolysate produced by a composite process and application thereof in shrimp culture; the algae hydrolysate is prepared from Ascophyllum nodosum by cutting, pulverizing, hydrolyzing and filtering; the alginic acid and the betaine produced by the process of the invention respectively have the following contents: the process is not less than 45g/l and not less than 26g/l, the process is not subjected to strong acid and strong alkali high-temperature treatment and high-temperature concentration treatment, the active substances of the product can keep higher activity, and simultaneously reach higher content requirements, new innovation is realized from production cost and biological activity, the seaweed hydrolysis product is applied to prevention and treatment of diseases in the prawn culture process, the product basically solves the problem of abuse of prawn culture chemicals, and the product is better than the effect of chemical prevention and treatment, and particularly has better effect on early prevention.)

1. A seaweed hydrolysate is characterized in that the seaweed hydrolysate is prepared by cutting, crushing, hydrolyzing and filtering leaf-soaked algae;

the preparation method of the seaweed hydrolysate comprises the following specific steps:

s1, cutting the dried Ascophyllum nodosum into blocks to obtain Ascophyllum nodosum blocks;

s2, putting the soaked algae blocks into a grinder to be ground, and sieving the ground soaked algae blocks through a 80-mesh sieve to obtain soaked algae powder;

s3, mixing the sieved paulownia powder with tartaric acid powder 10: 1, mixing with clean water at 40 ℃ of 1: 1(W/V), and soaking for 1 hour at the constant temperature of 40 ℃ to obtain a mixed solution;

s4, placing the mixed solution into a plurality of square grooves with the length of 20cm, placing the square grooves into an ice house, and rapidly cooling to-18 ℃;

s5, taking out the frozen mixed solution, and deicing at room temperature;

s6, after deicing, putting the ice in an ice house to cool to-25 ℃;

s7, taking out and quickly putting into a hydrolysis tank at 50 ℃, and simultaneously keeping the temperature of the hydrolysis tank unchanged;

s8, adjusting the pH value to 4.5, adding pectinase, and hydrolyzing for 12 hours;

s9, adjusting the pH value to 5.5, heating the tank to 55 ℃, adding the composite cellulase, and hydrolyzing for 8 hours;

s10, keeping the temperature of the tank constant, adjusting the pH value to 7.0, adding protease, and hydrolyzing for 28 hours;

and S11, mixing the hydrolyzed seaweed with phosphoric acid and citric acid, adjusting the pH value to 4.0, and filtering to remove impurities to obtain the hydrolyzed seaweed product.

2. The hydrolyzed seaweed product according to claim 1, wherein the size of the Ascophyllum nodosum is 1.5-2.5cm in step S1.

3. The hydrolyzed seaweed product according to claim 1, wherein in step S2, the sieved powder of Ascophyllum nodosum and tartaric acid are mixed in a ratio of 10: 1, mixing with clear water at 40 ℃, and soaking the materials for 1 hour at 40 ℃.

4. The hydrolyzed seaweed product according to claim 1, wherein the square-shaped tank is placed in an ice bank to be rapidly cooled to-18 ℃ in step S4.

5. The hydrolyzed seaweed product according to claim 1, wherein the amount of the raw materials fed into the hydrolysis tank in step S7 is 11% (dry powder basis).

6. The hydrolyzed seaweed product according to claim 1, wherein the amount of pectinase added in step S8 is 10000U/g pectinase 0.1% dry powder weight.

7. The hydrolyzed seaweed product according to claim 1, wherein the complex cellulase is added in an amount of 20000U/g cellulase 0.3% dry powder weight in step S9.

8. The hydrolyzed seaweed product according to claim 1, wherein the protease is added in an amount of 100000U/g protease 0.8% dry powder weight in step S10.

9. Use of a hydrolyzed seaweed product according to any one of claims l to 8 in shrimp farming.

Technical Field

The invention relates to the technical field of biology, in particular to a seaweed hydrolysate produced by a composite process and application thereof in shrimp culture.

Background

Alginic acid and betaine are main components in seaweed, are ideal bioactive substances in nature, and have good disease-resistant and bacteriostatic activities. Wherein, alginic acid has good blocking effect on replication process of baculovirus, polyhedrosis virus and other viruses, and betaine has good inhibition effect on bacterial diseases, particularly vibrio. The baculovirus and the vibrio are pathogenic microorganisms of common diseases in prawn culture at present, and the prevention and treatment effect on the main diseases in prawn culture can be better realized by inhibiting the propagation of the microorganisms. Alginic acid and betaine are mainly obtained by hydrolyzing seaweed, and the current seaweed hydrolysis process basically adopts physical, chemical, biological and other processes: 1. the physical method has low comprehensive utilization rate of the seaweed, high cost and difficult industrial production; 2. the chemical method has serious damage to active substances, and the obtained product has low activity and poor application effect; 3. the biological method overcomes the defects of the two methods, but the product concentration is low, the application is inconvenient, the cost is high, and if a concentration method is subsequently adopted, the active substances can be damaged at different degrees, and the cost is also increased. Therefore, the active concentration of the products developed by the seaweed is lower, the products are basically applied to the production of crops, the products are less applied to the breeding industry, and the hydrolysate of the seaweed is not directly applied to the disease control of prawns in literature report.

In summary, the research and development of a composite process for producing hydrolyzed seaweed and its application in shrimp culture still need to solve the key problems of high concentration and high activity in the field of biotechnology.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide a seaweed hydrolysate produced by a composite process and application thereof in shrimp culture, and the invention solves the problems by a specific process: 1. the problem of activity retention in the production process of alginic acid and betaine; 2. the concentration problem and the economic value problem of alginic acid and betaine; 3. the control problem of virus and vibrio diseases in the shrimp culture process is realized; 4. meanwhile, conditions are created for further application of the high-activity alginic acid product, the abuse problem of antibiotics and chemical drugs in prawn culture is solved, conditions are created for ecological culture, sustainable development and environmental ecological protection of prawn products, and meanwhile, a safer and more green food is provided for consumers to make effective guarantee.

In order to achieve the purpose, the invention provides the following technical scheme:

a hydrolyzed seaweed product is prepared from Ascophyllum nodosum by cutting, pulverizing, hydrolyzing and filtering;

the preparation method of the seaweed hydrolysate comprises the following specific steps:

s1, cutting the dried Ascophyllum nodosum into blocks to obtain Ascophyllum nodosum blocks;

s2, putting the soaked algae blocks into a grinder to be ground, and sieving the ground soaked algae blocks through a 80-mesh sieve to obtain soaked algae powder;

s3, mixing the sieved dunaliella powder and tartaric acid powder according to the weight ratio of 10: 1, mixing with clean water at 40 ℃ of 1: 1(W/V), and soaking for 1 hour at the constant temperature of 40 ℃ to obtain a mixed solution;

s4, placing the mixed solution into a plurality of square grooves with the length of 20cm, placing the square grooves into an ice house, and rapidly cooling to-18 ℃;

s5, taking out the frozen mixed solution, and deicing at room temperature;

s6, after deicing, putting the ice in an ice house to cool to-25 ℃;

s7, taking out and quickly putting into a hydrolysis tank at 50 ℃, and simultaneously keeping the temperature of the hydrolysis tank unchanged;

s8, adjusting the pH value to 4.5, adding pectinase, and hydrolyzing for 12 hours;

s9, adjusting the pH value to 5.5, heating the tank to 55 ℃, adding the composite cellulase, and hydrolyzing for 8 hours;

s10, keeping the temperature of the tank constant, adjusting the pH value to 7.0, adding protease, and hydrolyzing for 28 hours;

and S11, mixing the hydrolyzed seaweed with phosphoric acid and citric acid, adjusting the pH value to 4.0, and filtering to remove impurities to obtain the hydrolyzed seaweed product.

The invention is further configured to: in the step S1, the size of the Ascophyllum nodosum is 1.5-2.5 cm.

The invention is further configured to: in step S2, mixing the sieved paulownia powder and tartaric acid powder according to a ratio of 10: 1, mixing with clean water at 40 ℃ of 1: 1(W/V), and soaking at a constant temperature of 40 ℃ for 1 hour.

The invention is further configured to: in the step S4, the square groove is placed in an ice house to be rapidly cooled to-18 ℃, and taken out to be placed at room temperature for deicing.

The invention is further configured to: in the step S6, the ice is melted and then is put into an ice house to be cooled to-25 ℃;

the invention is further configured to: in step S7, the amount of feed in the hydrolysis tank is 11%.

The invention is further configured to: in the step S8, the addition amount of the pectinase is 10000U/g, and the weight of the pectinase is 0.1% of the dry powder.

The invention is further configured to: in the step S9, the addition amount of the complex cellulase is 20000U/g cellulase 0.3% dry powder weight.

The invention is further configured to: in step S10, the protease is added in an amount of 100000U/g protease 0.8% by weight of dry powder.

The invention provides an application of a seaweed hydrolysate in shrimp culture.

Advantageous effects

Compared with the known public technology, the technical scheme provided by the invention has the following beneficial effects:

(1) the alginic acid and the betaine produced by the process of the invention respectively have the following contents: the process has the advantages that the concentration is not less than 45g/l and not less than 26g/l, the process is not subjected to high-temperature treatment of strong acid and strong alkali and high-temperature concentration, the active substance of the product can keep higher activity, the higher content requirement is met, and new innovation is realized from the production cost and the biological activity.

(2) The application of the seaweed hydrolysate in the prevention and treatment of diseases in the process of prawn culture basically solves the problem of abuse of prawn culture chemicals, has better effect than the effect of chemical prevention and treatment, and particularly has better effect on early prevention.

Drawings

FIG. 1 is a flow chart of a composite process for producing hydrolyzed seaweed products.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The present invention will be further described with reference to the following examples.

Examples

Referring to fig. 1, a hydrolyzed seaweed product is prepared by cutting, crushing, hydrolyzing and filtering Ascophyllum nodosum;

the preparation method of the seaweed hydrolysate comprises the following specific steps:

step one, cutting the dried Ascophyllum nodosum into pieces, wherein the size of the Ascophyllum nodosum pieces is 1.5-2.5cm, and obtaining the Ascophyllum nodosum pieces.

And step two, putting the soaked leaf algae blocks into a grinder for grinding, and sieving by a 80-mesh sieve to obtain the soaked leaf algae powder.

Step three, mixing the sieved dunaliella powder and tartaric acid powder according to the ratio of 10: 1, mixing with clean water at 40 ℃ of 1: 1(W/V), and soaking for 1 hour at the constant temperature of 40 ℃ to obtain a mixed solution.

And step four, placing the mixed solution into a plurality of square grooves with the length of 20cm, placing the square grooves into an ice house, and rapidly cooling to-18 ℃.

And step five, taking out the frozen mixed solution, and deicing at room temperature.

And step six, after deicing, putting the ice into an ice house to cool to-25 ℃.

And seventhly, taking out and quickly putting into a hydrolysis tank at 50 ℃, and keeping the temperature of the hydrolysis tank unchanged, wherein the feeding amount (in terms of dry powder) in the hydrolysis tank is 11%.

And step eight, adjusting the pH value to 4.5, adding pectinase, and hydrolyzing for 12 hours, wherein the addition amount of the pectinase is 10000U/g of 0.1 percent of dry powder weight of the pectinase.

Ninth, adjusting the pH to 5.5, heating the tank to 55 ℃, adding the composite cellulase, and hydrolyzing for 8 hours, wherein the addition amount of the composite cellulase is 20000U/g cellulase 0.3% of the weight of dry powder;

step ten, keeping the tank temperature unchanged, adjusting the pH value to 7.0, adding protease, and hydrolyzing for 28 hours, wherein the addition amount of the protease is 100000U/g protease 0.8 percent of the weight of dry powder.

Step eleven, mixing phosphoric acid and citric acid after hydrolysis to adjust the pH value to 4.0, and filtering and removing impurities to obtain the seaweed hydrolysis product.

The invention provides an application of a seaweed hydrolysate in shrimp culture.

The application of the product in prawn culture comprises the following steps:

a. the use method of the preventive measures comprises the following steps: 1 liter of seaweed hydrolysis product is used once in a shrimp pond with the depth of 1.5 meters per mu, namely 1000ml of seaweed hydrolysis product is used in about 1000 tons of water, the alginic acid product is put in the shrimp fry in one season generally, the alginic acid product is applied together when the shrimp fry are put in the first time, and the alginic acid product is put in the shrimp fry when the shrimp fry are cultivated for one month in the second time;

b. the method of use in the early stages of the onset of the condition: if the prawn pond is in a swimming state and the food intake is reduced in the culture process, the prawn pond is indicated to be possibly damaged, and the prawn is fished through a net to observe whether the prawn is damaged or not, 2000 ml of seaweed hydrolysate can be applied to the prawn pond with the depth of 1.5 m per mu at the moment, 1000ml of seaweed hydrolysate can be used after one week, so that the disease can be basically prevented and treated, and 1000ml of seaweed hydrolysate can be used once again in the third week if the prawn pond is seriously damaged in the initial stage.

The application of the seaweed hydrolysate in the prevention and treatment of diseases in the process of prawn culture basically solves the problem of abuse of prawn culture chemicals, has better effect than the effect of chemical prevention and treatment, and particularly has better effect on early prevention.

Effect test

The detection method comprises the following steps: alginic acid adopts N/T3174-2017 standard m-hydroxybiphenyl method, and betaine adopts liquid chromatography.

According to the morbidity of the shrimp ponds in the past years, a plurality of shrimp ponds which have suffered from virus diseases and vibrio diseases in the past years are selected for carrying out comparison tests, the control tests are divided into four groups, namely a group A, a group B, a group C and a group D, each group is repeated by 3 ponds, and each pond is about 300 square meters.

Group A: is a traditional conventional drug control measure.

0.2 mg/L sodium dichloro isocyanurate is sprinkled in the whole pool, and 30-50 kg zeolite powder is put in each mu.

Group B: is a preventive measure for the seaweed hydrolysate.

When young shrimps are put in, 1000ml of seaweed hydrolysate is used in each acre of shrimp pond (1.5 m deep), and 1000ml of seaweed hydrolysate is used after one month.

Group C: is a routine medication measure for the initial onset of disease.

When the prawn has pond swimming phenomenon and the food intake is reduced, which may be the initial stage of disease, 0.3% streptomycin or florfenicol is added into the feed for alternate use, and the medicine is taken once in five days.

Group D: the seaweed hydrolysate is used for the early onset of disease.

2000 ml of seaweed hydrolysate is used in each acre of shrimp pond, and 1000ml of seaweed hydrolysate is used after one week.

The shrimp larvae in each shrimp pond are fed at a density of 0.8 ten thousand tails, the feed feeding and the shrimp pond management are kept unchanged from the prior art, and the test results are shown in table 1.

TABLE 1 test results

Note: the yield of each shrimp pond (unit: jin).

Through analyzing the relevant data in the table, the B group plan increases 15 jin of production compared with the A group, and increases 4% of production, which indicates that the B group achieves the expected prevention effect and realizes the production increase compared with the traditional prevention; compared with the group C, the group D increases the yield by 32.4 jin and 9.5%, has more obvious yield increasing effect, shows that the seaweed hydrolysate has certain treatment effect on the diseases of the prawns, also obtains more obvious yield increasing effect, and simultaneously shows that the chemical drug has better treatment effect on the diseases of the prawns, but has certain bad effect on the later-stage production of the prawns, and the yield is obviously inferior to the biological prevention and treatment scheme. Moreover, long-term application of antibiotics and other chemical drugs can also generate the drug resistance phenomenon of pathogenic bacteria and the generation of super pathogenic bacteria, and have greater negative effects on ecological breeding. The seaweed hydrolysis product has the advantages of no three wastes in the production process, mild process, capability of further fermenting the generated seaweed residues as a byproduct to produce an organic fertilizer product, environmental friendliness and low consumption, and accordance with the national green development requirement. The invention realizes the disease control of prawns by utilizing alginic acid and betaine in seaweed, is an innovation for ecological breeding of prawns, and provides a beneficial reference for other aquaculture. Therefore, the seaweed hydrolysate produced by the composite process has wider market prospect in shrimp culture and is more suitable for popularization.

The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.