阅读说明：本技术 一种阻止凡纳滨对虾肝肠孢子虫垂直传播的方法 (Method for preventing Vertically spreading of liver and intestinal sporozoon of Litopenaeus vannamei ) 是由 林海川 谢阳 陈金坤 于 2020-12-31 设计创作，主要内容包括：本发明公开了一种阻止凡纳滨对虾肝肠孢子虫垂直传播的方法包括以下步骤：亲虾的选择与暂养、亲虾的感染与强化、亲虾产卵、收集卵子、洗卵、洗幼体。本发明利用H试剂并结合采用特制的清洗系统装置,在对虾育苗初始阶段即对该胞子虫病害进行全面彻底的有效清除,阻断该胞子虫病害在对虾发育生长过程中的垂直传播,解决凡纳滨对虾增养殖中无法有效防控该胞子虫病害的难题。(The invention discloses a method for preventing Vertically propagating liver enterosporidium of Litopenaeus vannamei, which comprises the following steps: selecting and temporarily breeding parent shrimps, infecting and strengthening the parent shrimps, enabling the parent shrimps to lay eggs, collecting eggs, washing the eggs and washing larvae. The invention utilizes the H reagent and combines with a special cleaning system device, completely and effectively eliminates the cytozoosis disease in the initial stage of prawn breeding, blocks the vertical propagation of the cytozoosis disease in the prawn development and growth process, and solves the problem that the cytozoosis disease can not be effectively prevented and controlled in the litopenaeus vannamei breeding.)

1. A method for preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei is characterized by comprising the following steps:

step A: selecting and temporarily breeding parent shrimps, namely selecting healthy parent shrimps according to the ratio of male to female being 1: 2, wherein the temporarily breeding method of the parent shrimps comprises the following steps: putting the parent shrimps into a parent shrimp isolation pool, soaking the parent shrimps in NaOH solution with the pH value of more than or equal to 14 for 24 hours before the isolation pool is used, changing water for the whole isolation pool through a Dow water treater according to 40% every day, controlling the water temperature to be 28 ℃ and the salinity to be 28-34 per thousand during the whole temporary culture period, ensuring the dissolved oxygen to be more than 5mg/L and the pH value to be 8.4-8.7, and feeding the parent shrimps with the total ammonia nitrogen content of less than 0.6mg/L according to 2% of the weight of the parent shrimps every day for 4 times every day;

and B: infection and enhancement of parent shrimps: after the parent shrimps are temporarily cultured in the isolation pond for 10 days, feeding the parent shrimps mixed with the shrimp liver intestine cytozoon liquid, wherein the feeding amount is 2 percent of the weight, and after the parent shrimps are continuously fed for 15 days, the parent shrimps are proved to be positive by PCR detection; taking parent shrimps in a parent shrimp isolation pond, cutting off eye handles of the parent shrimps in the parent shrimp isolation pond every other day after detection, changing the water change amount by 60% every day, controlling the illumination intensity at 500x, adopting the optimized feed to calculate the feeding amount of the feed every day according to 2% of the weight of the parent shrimps, feeding the parent shrimps for 4 times every day, and after 20 days of cultivation, enabling the size of the parent shrimps to reach 47-60g, enabling the gonads of the parent shrimps to be full, and enabling male shrimps to be mature through fine clamping.

And C: parent shrimps lay eggs: after the sexual maturity of the parent shrimps is reached, lightly picking out the parent shrimps with mature gonads of the parent shrimps in the parent shrimp isolation pond at night, checking whether mating is completed or not, putting the mated parent shrimps, namely male shrimps with fine clamps and firmness at the genitals of the parent shrimps, into a spawning isolation pond, wherein the water in the spawning isolation pond is clean water treated by a Dow processor, the water temperature is controlled at 29 ℃, the dissolved oxygen is more than 5mg/L, and the total ammonia nitrogen in the water is 0; placing the unmatched female shrimps back to the net cage of the parent shrimp isolation pond, taking out the net cage after picking up all female shrimps with full gonads in all the ponds, returning the un-inseminated female shrimps to the parent shrimp isolation pond, and picking up the female shrimps which are inseminated to count the tails when the night; slightly picking out female shrimps and parent shrimps from the spawning isolation pool at night;

step D: collecting the ovum: collecting eggs in an egg-laying pond by using a 300-mesh net cage and a 300-mesh fishing net in the morning every other day, and putting the collected parent shrimp eggs into a 20L white barrel; the water in the white barrel is clean water treated by a Dow's water treatment device, the water temperature is 29 ℃, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0; placing micropore aeration in a white barrel, adjusting air flow to a micro-boiling state, taking 10ml by using a 10ml liquid transfer dropper, placing the 10ml liquid transfer dropper into a 100ml beaker, adding 90ml of clean ultrafiltration water into the beaker, taking 1ml by using a 1ml glass liquid transfer tube, and calculating the number of shrimp eggs in the liquid transfer tube, so that the total number of the shrimp eggs is 280-320 ten thousand;

step E: and (3) washing eggs: 10 cups of 500ml of water containing the shrimps are carefully taken from a white barrel by using a 500ml beaker, and egg collecting barrels in cleaning system devices with the numbers of 1-10 are randomly and respectively placed, so that the number of the shrimp eggs in each cleaning system device is 7-8 ten thousand; the water in the cleaning system device is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the dissolved oxygen is more than 5mg/L, the total ammonia nitrogen nitrite in the water is 0, 2ppm of H solution, namely 0.08ml of H reagent is added into the water, and the mixture is uniformly mixed; carefully fishing out macroscopic female shrimp feces and impurities by using a 60-mesh fishing net, adjusting the microporous oxygen increasing air stone in the egg collecting barrel to a slightly boiling state, and soaking shrimp eggs in the solution for 2 minutes; slowly adding prepared cleaning water into the egg collecting barrel after 2 minutes, wherein the cleaning water is clean water treated by a Dow water treater, the water temperature is controlled at 29 ℃, the pH value is 8.6, and the total ammonia nitrogen nitrite in the water is 0, so that the water in the cleaning system device is discharged while entering, the volume of the inflowing water is 20 times of that of the cleaning barrel, and the total amount of the inflowing cleaning water is 800L; after the eggs are washed, the egg collecting barrel is taken out gently, the eggs in the egg collecting barrel are transferred to an incubation barrel with the corresponding number of 1-10, water in the incubation barrel is clean water treated by a Dow water treater, the water level is 1m, the water temperature is 32 ℃, dissolved oxygen is more than 5mg/L, ammonia nitrogen and nitrite in the water are all 0, a micropore aerostone is placed in the incubation barrel, and the air flow is adjusted to be in a slightly boiling state;

step F: and (3) larva washing: checking that the shrimp eggs in the hatching barrel with the number of 1-10 are normally hatched every noon, placing a cleaning system device with the number of 11-20 below the hatching barrel, wherein water in the cleaning system device is clean water treated by a Dow water treater, the water temperature is controlled at 32 ℃, the PH is 8.6, the total ammonia nitrogen and nitrite in the water are all 0, closing the micropore aeroliths in the hatching barrel, and standing for 10 minutes; opening micropores in the egg collecting barrel after 10 minutes to increase oxygen, adjusting the air flow to slightly boil, opening a pvc ball valve below the hatching barrel, and adjusting the air flow to enable the water flow with the larvae to slowly flow into the corresponding egg collecting barrel in the cleaning system device until no water flows out; after the larvae are collected, taking 500ml of water out of a cleaning barrel by using a 500ml beaker, putting 0.08ml of H reagent, uniformly mixing, pouring the mixture into an egg collecting barrel, slightly shaking the egg collecting barrel up and down to uniformly distribute the solution in the cleaning barrel and the egg collecting barrel, opening prepared cleaning water, slowly flowing the cleaning water into the egg collecting barrel, controlling the water temperature to be 32 ℃, uniformly distributing the solution in the cleaning barrel and the egg collecting barrel, and discharging the water in a cleaning system device while feeding the water, wherein the volume of the inflowing water is 40 times that of the cleaning barrel, and the total volume of the inflowing water is 1600L of cleaning water; after the cleaning, collecting the larvae in the cleaning system device with the number of 11-20, conveying the larvae to an isolated seedling raising pond, and entering a conventional standard seedling raising program.

2. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: healthy parent shrimps in the step A. The healthy parent shrimps areThe body length is 16-18cm, the weight is 32-40g, the body is large, the body color is normal, the paraplegia is complete, the gill part is clean, and no specific virus infection exists; the parent shrimps are temporarily cultured according to the ratio of 11-14 tails/m²The parent shrimps are put into a parent shrimp isolation pond with the density of 25m²The depth of the cement pond is 0.6 m.

3. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: taking feces from a culture pond with outbreak of the shrimp liver intestine cytozoon, wherein the symptoms of the shrimp liver intestine cytozoon are obvious and the shrimp liver intestine cytozoon is positive through PCR detection, putting 0.1g of the shrimp feces with outbreak of the shrimp liver intestine cytozoon into a homogenizer, adding 1mL of LPBS buffer solution, carrying out bath grinding uniformly, putting the shrimp liver intestine cytozoon liquid into a 1.5mL of EP tube, centrifuging for 5min at 3000r/min, taking 200uL of supernatant, adding the supernatant into 1800uL of PBS buffer solution, filtering by using a filter which is sterilized at high temperature and is provided with 0.22uL of meshes, taking 500uL of filtrate, adding 49.5mL of PBS buffer solution with pH7.4, mixing uniformly, totaling 50mL, and storing at 4 ℃ for later use.

4. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: the optimized feed in the step B is prepared from parent shrimp feed, sandworms, squids, beef livers and the like according to the proportion of 1: 3: 1.

5. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: and C, the length, the width and the height of the net cage in the step C are 1m x 1m, the periphery of the net cage is surrounded by 40 meshes, the upper surface and the lower surface of the net cage are emptied, the net cage is kept quiet when spawning is carried out, all lights are turned off, and the spawning rate is improved.

6. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: the cleaning system device in the step E and the step F comprises a cleaning barrel and an egg collecting barrel, wherein the cleaning barrel is a square plastic barrel with the length of 40cm, the width of 40cm and the height of 25 cm; the egg collecting barrel is a circular plastic barrel with the radius of 15cm and the height of 35cm, a rectangular ring is cut at a position 10cm away from the barrel bottom, the cut holes are 40cm long and 30cm wide, and a 250-mesh filter screen is fixed in each hole.

7. The method of claim 6, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: in the use process of the egg collecting barrel, the egg collecting barrel is placed in the cleaning barrel, water is added into the egg collecting barrel, the water can carry tiny excrement, impurities and shrimp liver enterozoon to flow out of the filter screen, a micropore oxygen increasing air stone is placed in the egg collecting barrel, and the air quantity is adjusted to slightly boil.

8. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: and the reagent H in the step E and the step F is prepared from anionic polyacrylamide, povidone iodine and purified water according to the ratio of 1: 5: 10.

9. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: in the step E and the step F, the hatching barrel is a plastic barrel with the bottom radius of 25cm and the height of 1.5m, a pvc pipe with the caliber of 32mm is arranged at the barrel bottom, a ball valve assembled at a water outlet is 15cm along with a plastic hose with the caliber of 32mm, and an elbow and the pvc pipe with the caliber of 32mm and the height of 20cm are assembled in the barrel.

10. The method of claim 1, wherein the method comprises the step of preventing the vertical transmission of the liver enterosporidium of the litopenaeus vannamei: and E, installing a 50w incandescent lamp at a position 40cm above the hatching barrel, keeping the incandescent lamp in a normally-on state during hatching, and placing the hatching barrel on a table top 35cm above the ground.

Technical Field

The invention relates to a method for preventing the vertical propagation of litopenaeus vannamei liver enterosporidium, belongs to the field of preventing propagation of aquaculture diseases, and particularly relates to a method for preventing and controlling the vertical propagation of the litopenaeus vannamei liver enterosporidium by adopting preparations such as anionic polyacrylamide, povidone iodine and the like.

Background

The litopenaeus vannamei is a common aquatic product prawn culture species in China. The shrimp liver intestine cytozoon (EPH) of the litopenaeus vannamei is a cytozoon disease which seriously harms the normal growth of the litopenaeus vannamei, has extremely strong reproductive capacity, and is difficult to achieve the aim of completely killing the litopenaeus vannamei in the breeding environment by a conventional mode (acid or alkaline solution).

Disclosure of Invention

The invention aims to solve the technical problem of providing a method for preventing the vertical propagation of the enterosporidium of the litopenaeus vannamei.e. the preparation agents such as anionic polyacrylamide, povidone iodine and the like are adopted to prevent and control the propagation of the enterosporidium of the litopenaeus vannamei.

In order to solve the technical problems, the technical scheme of the invention is as follows: the innovation point of the method for preventing the liver and intestinal sporozoon of the litopenaeus vannamei from vertically spreading is that the method comprises the following steps:

step A: selecting and temporarily breeding parent shrimps, namely selecting healthy parent shrimps according to the ratio of male to female being 1: 2, wherein the temporarily breeding method of the parent shrimps comprises the following steps: putting the parent shrimps into a parent shrimp isolation pool, soaking the parent shrimps in NaOH solution with the pH value of more than or equal to 14 for 24 hours before the isolation pool is used, changing water for the whole isolation pool through a Dow water treater according to 40% every day, controlling the water temperature to be 28 ℃ and the salinity to be 28-34 per thousand during the whole temporary culture period, ensuring the dissolved oxygen to be more than 5mg/L and the pH value to be 8.4-8.7, and feeding the parent shrimps with the total ammonia nitrogen content of less than 0.6mg/L according to 2% of the weight of the parent shrimps every day for 4 times every day;

and B: infection and enhancement of parent shrimps: after the parent shrimps are temporarily cultured in the isolation pond for 10 days, feeding the parent shrimps mixed with the shrimp liver intestine cytozoon liquid, wherein the mixed feed amount is 10ml/kg, the feeding amount is 2 percent of the body weight, and after the parent shrimps are continuously fed for 15 days, the parent shrimps are proved to be positive by PCR detection; taking parent shrimps in a parent shrimp isolation pond, cutting off eye handles of the parent shrimps in the parent shrimp isolation pond every other day after detection, changing the water change amount by 60% every day, controlling the illumination intensity at 500x, adopting the optimized feed to calculate the feeding amount of the feed every day according to 2% of the weight of the parent shrimps, feeding the parent shrimps for 4 times every day, and after 20 days of cultivation, enabling the size of the parent shrimps to reach 47-60g, enabling the gonads of the parent shrimps to be full, and enabling male shrimps to be mature through fine clamping.

And C: parent shrimps lay eggs: after the sexual maturity of the parent shrimps is reached, lightly picking out the parent shrimps with mature gonads of the parent shrimps in the parent shrimp isolation pond at night, checking whether mating is completed or not, putting the mated parent shrimps, namely male shrimps with fine clamps and firmness at the genitals of the parent shrimps, into a spawning isolation pond, wherein the water in the spawning isolation pond is clean water treated by a Dow processor, the water temperature is controlled at 29 ℃, the dissolved oxygen is more than 5mg/L, and the total ammonia nitrogen in the water is 0; placing the unmatched female shrimps back to the net cage of the parent shrimp isolation pond, taking out the net cage after picking up all female shrimps with full gonads in all the ponds, returning the un-inseminated female shrimps to the parent shrimp isolation pond, and picking up the female shrimps which are inseminated to count the tails when the night; slightly picking out female shrimps and parent shrimps from the spawning isolation pool at night;

step D: collecting the ovum: collecting eggs in an egg-laying pond by using a 300-mesh net cage and a 300-mesh fishing net in the morning every other day, and putting the collected parent shrimp eggs into a 20L white barrel; the water in the white barrel is clean water treated by a Dow's water treatment device, the water temperature is 29 ℃, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0; placing micropore aeration in a white barrel, adjusting air flow to a micro-boiling state, taking 10ml by using a 10ml liquid transfer dropper, placing the 10ml liquid transfer dropper into a 100ml beaker, adding 90ml of clean ultrafiltration water into the beaker, taking 1ml by using a 1ml glass liquid transfer tube, and calculating the number of shrimp eggs in the liquid transfer tube, so that the total number of the shrimp eggs is 280-320 ten thousand;

step E: and (3) washing eggs: 10 cups of 500ml of water containing the shrimps are carefully taken from a white barrel by using a 500ml beaker, and egg collecting barrels in cleaning system devices with the numbers of 1-10 are randomly and respectively placed, so that the number of the shrimp eggs in each cleaning system device is 7-8 ten thousand; the water in the cleaning system device is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the dissolved oxygen is more than 5mg/L, the total ammonia nitrogen nitrite in the water is 0, 2ppm of H solution, namely 0.08ml of H reagent is added into the water, and the mixture is uniformly mixed; carefully fishing out macroscopic female shrimp feces and impurities by using a 60-mesh fishing net, adjusting the microporous oxygen increasing air stone in the egg collecting barrel to a slightly boiling state, and soaking shrimp eggs in the solution for 2 minutes; slowly adding prepared cleaning water into the egg collecting barrel after 2 minutes, wherein the cleaning water is clean water treated by a Dow water treater, the water temperature is controlled at 29 ℃, the pH value is 8.6, and the total ammonia nitrogen nitrite in the water is 0, so that the water in the cleaning system device is discharged while entering, the volume of the inflowing water is 20 times of that of the cleaning barrel, and the total amount of the inflowing cleaning water is 800L; after the eggs are washed, the egg collecting barrel is taken out gently, the eggs in the egg collecting barrel are transferred to an incubation barrel with the corresponding number of 1-10, water in the incubation barrel is clean water treated by a Dow water treater, the water level is 1m, the water temperature is 32 ℃, dissolved oxygen is more than 5mg/L, ammonia nitrogen and nitrite in the water are all 0, a micropore aerostone is placed in the incubation barrel, and the air flow is adjusted to be in a slightly boiling state;

step F: and (3) larva washing: checking that the shrimp eggs in the hatching barrel with the number of 1-10 are normally hatched every noon, placing a cleaning system device with the number of 11-20 below the hatching barrel, wherein water in the cleaning system device is clean water treated by a Dow water treater, the water temperature is controlled at 32 ℃, the PH is 8.6, the total ammonia nitrogen and nitrite in the water are all 0, closing the micropore aeroliths in the hatching barrel, and standing for 10 minutes; opening micropores in the egg collecting barrel after 10 minutes to increase oxygen, adjusting the air flow to slightly boil, opening a pvc ball valve below the hatching barrel, and adjusting the air flow to enable the water flow with the larvae to slowly flow into the corresponding egg collecting barrel in the cleaning system device until no water flows out; after the larvae are collected, taking 500ml of water out of a cleaning barrel by using a 500ml beaker, putting 0.08ml of H reagent, uniformly mixing, pouring the mixture into an egg collecting barrel, slightly shaking the egg collecting barrel up and down to uniformly distribute the solution in the cleaning barrel and the egg collecting barrel, opening prepared cleaning water, slowly flowing the cleaning water into the egg collecting barrel, controlling the water temperature to be 32 ℃, uniformly distributing the solution in the cleaning barrel and the egg collecting barrel, and discharging the water in a cleaning system device while feeding the water, wherein the volume of the inflowing water is 40 times that of the cleaning barrel, and the total volume of the inflowing water is 1600L of cleaning water; after the cleaning, collecting the larvae in the cleaning system device with the number of 11-20, conveying the larvae to an isolated seedling raising pond, and entering a conventional standard seedling raising program.

Preferably, healthy parent shrimps are used in step A. The healthy parent shrimps are 16-18cm in body length, 32-40g in body weight, large in body size, normal in body color, complete in paraplegia, clean in gill part and free of specific virus infection; the parent shrimps are temporarily cultured according to the ratio of 11-14 tails/m²The parent shrimps are put into a parent shrimp isolation pond with the density of 25m²The depth of the cement pond is 0.6 m.

Preferably, the preparation method of the shrimp liver intestine cytozoon liquid in the step B includes the steps of taking excrement from a culture pond with outbreak of shrimp liver intestine cytozoon, wherein the symptoms of the shrimp liver intestine cytozoon in the pond are obvious and the shrimp liver intestine cytozoon is positive through PCR detection, putting 0.1g of the excrement of the shrimp with outbreak of shrimp liver intestine cytozoon into a homogenizer, adding 1mL of LPBS buffer solution, carrying out bath grinding uniformly, putting the shrimp liver intestine cytozoon liquid into a 1.5mL EP tube, centrifuging for 5min at 3000r/min, taking 200uL of supernatant, adding the supernatant into 1800uL of PBS buffer solution, filtering the supernatant by a filter which is sterilized at high temperature and is provided with 0.22uL of meshes, taking 500uL of filtrate, adding the filtrate into 49.5mL of PBS buffer solution with pH7.4, mixing uniformly, totaling 50mL, and storing the filtrate at 4 ℃ for later use.

Preferably, the optimized feed in the step B is parent shrimp feed, sandworm, squid, beef liver and the like which are mixed according to the proportion of 1: 3: 1.

Preferably, in the step C, the length and width of the net cage are 1m x 1m, the periphery of the net cage is surrounded by 40 meshes, the upper surface and the lower surface of the net cage are emptied, the net cage is kept quiet during egg laying, all lights are turned off, and the egg laying rate is improved.

Preferably, the cleaning system device in the step E and the step F comprises a cleaning barrel and an egg collecting barrel, wherein the cleaning barrel is a square plastic barrel with a length of 40cm, a width of 40cm and a height of 25 cm; the egg collecting barrel is a circular plastic barrel with the radius of 15cm and the height of 35cm, a rectangular ring is cut at a position 10cm away from the barrel bottom, the cut holes are 40cm long and 30cm wide, and a 250-mesh filter screen is fixed in each hole.

Preferably, in the using process of the egg collecting barrel, the egg collecting barrel is placed in the cleaning barrel, water is added into the egg collecting barrel, the water can carry fine excrement, impurities and shrimp liver enterozoon to flow out of the filter screen, a micropore aeration stone is placed in the egg collecting barrel, and the air volume is adjusted to slightly boil.

Preferably, the reagent H in the step E and the step F is prepared from anionic polyacrylamide, povidone iodine and purified water according to the ratio of 1: 5: 10.

Preferably, the hatching barrel in the step E and the step F is a plastic barrel with the bottom radius of 25cm and the height of 1.5m, a pvc pipe with the caliber of 32mm is arranged at the bottom of the barrel, a ball valve assembled at a water outlet is 15cm along with a plastic hose with the caliber of 32mm, and an elbow and the pvc pipe with the caliber of 20cm and the height of 32mm are assembled in the barrel.

Preferably, a 50w incandescent lamp is arranged at a position 40cm above the hatching barrel in the step E, the incandescent lamp keeps a normally-on state during hatching, and the hatching barrel is placed on a table top 35cm above the ground.

The invention has the beneficial effects that: the invention utilizes the H reagent and combines with a special cleaning system device, completely and effectively eliminates the cytozoosis disease in the initial stage of prawn breeding, blocks the vertical propagation of the cytozoosis disease in the prawn development and growth process, and solves the problem that the cytozoosis disease can not be effectively prevented and controlled in the litopenaeus vannamei breeding.

Detailed Description

The present invention will be described in further detail with reference to examples.

Example 1

1) Selecting and temporarily culturing parent shrimps: selecting healthy parent shrimps according to the ratio of male to female of 1: 2, wherein the healthy parent shrimps have the body length of 17cm, the body weight of 36g, large body size, normal body color, complete paraplegia, clean gill parts and no specific virus infection. The method for temporarily breeding the parent shrimps comprises the following steps: according to 12 tails/m²The parent shrimps are put into a parent shrimp isolation pond with the size of 25m²The cement pond is 0.6m deep, the isolation pond is soaked in NaOH solution with pH being more than or equal to 14 for 24 hours before use, water in the whole isolation field passes through a Dow water treater, the filtration pore diameter is 0.1-0.2 um, water is changed according to 40% every day, the water temperature is controlled at 28 ℃ during the whole temporary culture period, the salinity is 28-34 per mill, dissolved oxygen is ensured to be more than 5mg/L, the pH is 8.6, the content of total ammonia nitrogen in water is less than 0.6mg/L, parent shrimp feed is fed according to 2% of the weight of the shrimps every day, and the parent shrimp feed is fed for 4 times every day.

2) Infection and enhancement of parent shrimps: after the parent shrimps are temporarily raised in the isolation field for 10 days, feeding the parent shrimps with the shrimp liver intestine cytozoon liquid, wherein the amount of the mixed feed is 10ml/kg, the feeding amount is 2 percent of the body weight, and after the parent shrimps are continuously fed for 15 days, taking fresh excrement produced by the parent shrimps in the isolation pool of the parent shrimps, and the excrement is proved to be positive by PCR detection. And (3) after the detection, removing the eye handle of the female shrimp in the parent shrimp isolation pond every other day, changing the water amount by 60% every day, and controlling the illumination intensity at 500 x. Feeding parent shrimp with feed, sandworm, Loligo chinensis Gray, and hepar bovis Seu Bubali at ratio of 1: 3: 1. The feeding amount is calculated according to 2 percent of the weight of the parent shrimps every day, and the feeding is carried out for 4 times every day. After 20 days of cultivation, the size of the parent shrimps reaches 50 g. The gonads of the female shrimps are full, and the male shrimps are mature.

The preparation method of the shrimp liver intestine cytozoon liquid comprises the following steps: taking excrement from the culture pond with the outbreak of the shrimp liver enterosporidium, the shrimp pond has obvious liver enterosporidium symptom, and PCR detection proves that the shrimp liver enterosporidium is positive. Putting 0.1g of prawn feces of outbreak of shrimp liver enterocytozoon into a homogenizer, adding 1mL of LPBS buffer solution, carrying out bath grinding uniformly, then putting the shrimp liver enterocytozoon liquid into a 1.5mL EP tube, centrifuging for 5min at 3000r/min, taking 200uL of supernatant, adding into 1800uL of PBS buffer solution, and filtering by using a filter which is sterilized at high temperature and is provided with 0.22uL of meshes. 500uL of the filtrate was added to 49.5mL of PBS buffer (pH 7.4), and the mixture was mixed, totaling 50mL, and stored at 4 ℃ until use.

3) Spawning parent shrimps and collecting ova: after the sexual maturity of the parent shrimps is reached, lightly picking out the mature parent shrimps of the gonads of the parent shrimps in the parent shrimp isolation pond at 7 o' clock evening, checking whether mating is completed or not, placing the mated parent shrimps, namely male shrimps, into the spawning isolation pond firmly by finely clamping the female shrimps at the genitals, wherein the spawning isolation pond is a cement pond with an area of 25 square meters and a water level of 0.6m, water is clean water treated by a Dow processor, the water temperature is controlled at 29 ℃, the pH is 8.6, dissolved oxygen is more than 5mg/L, and the total ammonia nitrogen in the water is 0. And (4) putting the unmatched female shrimps back to the net cage of the parent shrimp isolation pond, taking out the net cage after picking up all the female shrimps with full gonads in all the ponds, and returning the un-inseminated female shrimps to the parent shrimp isolation pond. And selecting 15 tails of the female shrimps which are qualified for insemination at night. The length, width and height of the net cage are 1m x 1m, the periphery of the net cage is surrounded by 40 meshes, and the upper surface and the lower surface of the net cage are emptied. When spawning, the device keeps quiet, all lights are turned off, and the spawning rate is improved. And lightly picking out the female shrimps and the parent shrimp isolation pond from the spawning isolation pond at 12 pm. Collecting the eggs in the spawning and leaving pond by using a 300-mesh net cage and a 300-mesh fishing net at 1 point every other morning. The collected parent shrimp eggs were placed in a 20L white bucket. The water in the white barrel is clean water treated by a Dow water treatment device, the water temperature is 29 ℃, the pH value is 8.6, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0. Placing micropore aeration in a white barrel, adjusting air flow to a micro-boiling state, taking 10ml by using a 10ml liquid transfer dropper, putting the 10ml liquid transfer dropper into a 100ml beaker, adding 90ml of clean ultrafiltration water into the beaker, taking 1ml by using a 1ml glass liquid transfer tube, and calculating the tail 16 shrimp egg numbers in the liquid transfer tube, thereby obtaining the total shrimp egg number of 320 ten thousand.

4) And (3) washing eggs: 10 shrimp-containing 500ml water were carefully taken out of the white barrel in a 500ml beaker, and egg collecting barrels of 1 to 10 cleaning system units were randomly placed, respectively, and it was found that the number of shrimp eggs in each cleaning system unit was 8 ten thousand. The water in the cleaning system device is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the PH is 8.6, the dissolved oxygen is more than 5mg/L, the total ammonia nitrogen nitrite in the water is 0, 2ppm of H solution, namely 0.08ml of H reagent is added into the water, and the mixture is uniformly mixed. The macroscopic mother shrimp feces and impurities are carefully fished out by a 60-mesh fishing net, and the microporous oxygen increasing air stone in the egg collecting barrel is adjusted to be slightly boiled, so that the shrimp eggs are soaked in the solution for 2 minutes. After 2 minutes, the prepared rinsing water was slowly added to the egg collecting bucket. The cleaning water is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the pH value is 8.6, and the total ammonia nitrogen nitrite in the water is 0. The water in the washing system apparatus was discharged while being introduced, and the total volume of the inflow water was 20 times the volume of the washing tub, and 800L of the inflow water was counted. After the eggs are washed, the egg collecting barrel is taken out slightly, and the eggs in the egg collecting barrel are transferred to the hatching barrel with the corresponding number of 1-10. The water in the hatching barrel is clean water treated by a Dow water treater, the water level is 1m, the water temperature is 32 ℃, the PH value is 8.6, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0. A micropore aeration stone is placed in the hatching barrel, and the air quantity is adjusted to be in a slight boiling state. A50 w incandescent lamp is arranged at a position 40cm above the hatching barrel, and the incandescent lamp is kept in a normally-on state during hatching. The hatching barrel is placed on a table top 35cm away from the ground.

The H reagent is prepared from anionic polyacrylamide, povidone iodine and purified water according to the proportion of 1: 5: 10. The cleaning system device comprises two parts, wherein the first part is a cleaning barrel, and the cleaning barrel is a square plastic barrel with the length of 40cm, the width of 40cm and the height of 25 cm. The second part is an egg collecting barrel, the radius of the egg collecting barrel is 15cm, the height of the egg collecting barrel is 35cm, a rectangular ring-shaped cutting part is arranged at a position 10cm away from the barrel bottom, the cut holes are 40cm in length and 30cm in width, and a 250-mesh filter screen is fixed in each hole. In the use process, the egg collecting barrel is placed in the cleaning barrel, water is added into the egg collecting barrel, and the water can carry tiny excrement, impurities and shrimp liver enterozoon to flow out of the filter screen. A microporous oxygen increasing air stone is placed in the egg collecting barrel, and the air quantity is adjusted to slightly boil. The hatching barrel is a plastic barrel with the bottom radius of 25cm and the height of 1.5m, a pvc pipe with the caliber of 32mm is arranged at the bottom of the barrel, a ball valve assembled at a water outlet is 15cm along with a plastic hose with the caliber of 32mm, and an elbow and the pvc pipe with the caliber of 32mm with the height of 20cm are assembled inside the barrel.

5) And (3) larva washing: every 12 noon, the hatching barrels with the numbers of 1-10 are checked and all hatched normally. The cleaning system device with the number of 11-20 is placed below the hatching barrel. The water in the cleaning system device is clean water treated by a Dow water treater, the water temperature is controlled at 32 ℃, the PH is 8.6, and the total ammonia nitrogen nitrite in the water is 0. The hatching barrel is arranged on a table top 35cm higher than the ground. And (5) turning off the microporous aerator stone in the incubation barrel, and standing for 10 minutes. After 10 minutes, the micropores in the egg collecting barrel are opened to increase oxygen, and the air quantity is adjusted to slightly boil. And opening a pvc ball valve below the hatching barrel, and adjusting the size to ensure that the water flow with the larvae slowly flows into the egg collecting barrel in the corresponding cleaning system device until no water flows out. After collecting the larvae, taking 500ml of water out of the cleaning barrel by using a 500ml beaker, putting 0.08ml of H reagent, uniformly mixing, pouring into the egg collecting barrel, slightly shaking the egg collecting barrel up and down to uniformly distribute the solution in the cleaning barrel and the egg collecting barrel, opening the prepared cleaning water, and slowly flowing the cleaning water into the egg collecting barrel. The cleaning water is clean water treated by a Dow water treatment device, the water temperature is controlled at 32 ℃, the pH value is 8.6, and the total ammonia nitrogen nitrite in the water is 0. The water in the washing system apparatus was discharged while being introduced, and the total amount of the inflow water was 40 times the volume of the washing tub, and 1600L of the washing water was introduced. After the cleaning, collecting the larvae in the cleaning system device with the number of 11-20, conveying the larvae to an isolated seedling raising pond, and entering a conventional standard seedling raising program.

Example 2

1) Selecting and temporarily culturing parent shrimps: selecting healthy parent shrimp according to the ratio of male to female of 1: 2The shrimp has a body length of 16cm, a weight of 32g, a large body size, a normal body color, intact paraplegia, clean gill and no specific virus infection. The temporary parent shrimp breeding method comprises the following steps: according to 14 tails/m²The parent shrimps are put into a parent shrimp isolation pond with the size of 25m²The cement pond is 0.6m deep, the isolation pond is soaked in NaOH solution with pH being more than or equal to 14 for 24 hours before use, water in the whole isolation field passes through a Dow water treater, the filter pore diameter is 0.1-0.2 um, water change is carried out according to 40% every day, the water temperature is controlled at 28 ℃ during the whole temporary culture period, the salinity is 28-34 per mill, the dissolved oxygen is ensured to be more than 5mg/L, the pH is 8.4, and the total ammonia nitrogen content in the water is lower than 0.6 mg/L. Feeding parent shrimp feed according to 2% of the weight of the shrimp every day, and feeding for 4 times every day.

2) Infection and enhancement of parent shrimps: after the parent shrimps are temporarily cultured in the isolation area for 10 days, the parent shrimps are fed with the parent shrimp feed mixed with the shrimp liver enterozoon liquid, the mixed feed amount is 10ml/kg, and the feeding amount is 2 percent of the body weight. After 15 days of continuous feeding, fresh excrement produced by parent shrimps in the parent shrimp isolation pool is taken and proved to be positive by PCR detection. And (3) after the detection, removing the eye handle of the female shrimp in the parent shrimp isolation pond every other day, changing the water amount by 60% every day, and controlling the illumination intensity at 500 x. Feeding parent shrimp with feed, sandworm, Loligo chinensis Gray, and hepar bovis Seu Bubali at ratio of 1: 3: 1. The feeding amount is calculated according to 2 percent of the weight of the parent shrimps every day, and the feeding is carried out for 4 times every day. After 20 days of cultivation, the size of the parent shrimps reaches 47 g. The gonads of the female shrimps are full, and the male shrimps are mature.

The preparation method of the shrimp liver intestine cytozoon liquid comprises the following steps: taking excrement from the culture pond with the outbreak of the shrimp liver enterosporidium, the shrimp pond has obvious liver enterosporidium symptom, and PCR detection proves that the shrimp liver enterosporidium is positive. Putting 0.1g of prawn feces of outbreak of shrimp liver enterocytozoon into a homogenizer, quickly adding 1mL of LPBS buffer solution, carrying out bath grinding uniformly, then putting the shrimp liver enterocytozoon liquid into a 1.5mL EP tube, centrifuging for 5min at 3000r/min, taking 200uL of supernatant, adding into 1800uL of PBS buffer solution, and filtering by using a filter which is sterilized at high temperature and is provided with 0.22uL of meshes. 500uL of the filtrate was added to 49.5mL of PBS buffer (pH 7.4), and the mixture was mixed, totaling 50mL, and stored at 4 ℃ until use.

3) Spawning parent shrimps and collecting ova: after the sexual maturity of the parent shrimps is reached, lightly picking out the mature parent shrimps of the gonads of the parent shrimps in the parent shrimp isolation pond at 7 o' clock evening, checking whether mating is completed or not, placing the mated parent shrimps, namely male shrimps, into the spawning isolation pond firmly by finely clamping the female shrimps at the genitals, wherein the spawning isolation pond is a cement pond with an area of 25 square meters and a water level of 0.6m, water is clean water treated by a Dow processor, the water temperature is controlled at 29 ℃, the pH is 8.4, the dissolved oxygen is more than 5mg/L, and the total ammonia nitrogen in the water is 0. And (4) putting the unmatched female shrimps back to the net cage of the parent shrimp isolation pond, taking out the net cage after picking up all the female shrimps with full gonads in all the ponds, and returning the un-inseminated female shrimps to the parent shrimp isolation pond. And selecting 17 tails of the female shrimps which are qualified for insemination at night. The length, width and height of the net cage are 1m x 1m, the periphery of the net cage is surrounded by 40 meshes, and the upper surface and the lower surface of the net cage are emptied. When spawning, the device keeps quiet, all lights are turned off, and the spawning rate is improved. And lightly picking out the female shrimps and the parent shrimp isolation pond from the spawning isolation pond at 12 pm. Collecting the eggs in the spawning and leaving pond by using a 300-mesh net cage and a 300-mesh fishing net at 1 point every other morning. The collected parent shrimp eggs were placed in a 20L white bucket. The water in the white barrel is clean water treated by a Dow water treatment device, the water temperature is 29 ℃, the pH value is 8.6, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0. Placing micropore aeration in a white barrel, adjusting air flow to a micro-boiling state, taking 10ml by using a 10ml liquid transfer dropper, putting the 10ml liquid transfer dropper into a 100ml beaker, adding 90ml of clean ultrafiltration water into the beaker, taking 1ml by using a 1ml glass liquid transfer tube, and calculating the tail 13 shrimp egg numbers in the liquid transfer tube, thereby obtaining the total number of the shrimp eggs of 280 ten thousand.

4) And (3) washing eggs: 10 shrimp-containing 500ml water were carefully taken out of the white barrel using a 500ml beaker, and egg collecting barrels of 1 to 10-numbered washing system apparatuses were randomly placed, respectively, and it was found that the number of shrimp eggs in each washing system apparatus was 7 ten thousand. The cleaning system device in the cleaning barrel comprises a buckle cleaning barrel and an ovum collecting barrel. The water in the cleaning system device is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the PH is 8.4, the dissolved oxygen is more than 5mg/L, the total ammonia nitrogen nitrite in the water is 0, 2ppm of H solution, namely 0.08ml of H reagent is added into the water, and the mixture is uniformly mixed. The macroscopic mother shrimp feces and impurities are carefully fished out by a 60-mesh fishing net, and the microporous oxygen increasing air stone in the egg collecting barrel is adjusted to be slightly boiled, so that the shrimp eggs are soaked in the solution for 2 minutes. After 2 minutes, the prepared rinsing water was slowly added to the egg collecting bucket. The cleaning water is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the pH value is 8.4, and the total ammonia nitrogen nitrite in the water is 0. The water in the washing system apparatus was discharged while being introduced, and the total volume of the inflow water was 20 times the volume of the washing tub, and 800L of the inflow water was counted. After the eggs are washed, the egg collecting barrel is taken out slightly, and the eggs in the egg collecting barrel are transferred to the hatching barrel with the corresponding number of 1-10. The water in the hatching barrel is clean water treated by a Dow water treater, the water level is 1m, the water temperature is 32 ℃, the PH value is 8.4, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0. A micropore aeration stone is placed in the hatching barrel, and the air quantity is adjusted to be in a slight boiling state. A50 w incandescent lamp is arranged at a position 40cm above the hatching barrel, and the incandescent lamp is kept in a normally-on state during hatching. The hatching barrel is placed on a table top 35cm away from the ground.

The H reagent is: the anionic polyacrylamide, the povidone iodine and the purified water are prepared according to the proportion of 1: 5: 10. The cleaning system device comprises two parts, wherein the first part is a cleaning barrel, and the cleaning barrel is a square plastic barrel with the length of 40cm, the width of 40cm and the height of 25 cm. The second part is an egg collecting barrel, the radius of the egg collecting barrel is 15cm, the height of the egg collecting barrel is 35cm, a rectangular ring-shaped cutting part is arranged at a position 10cm away from the barrel bottom, the cut holes are 40cm in length and 30cm in width, and a 250-mesh filter screen is fixed in each hole. In the use process, the egg collecting barrel is placed in the cleaning barrel, water is added into the egg collecting barrel, and the water can carry tiny excrement, impurities and shrimp liver enterozoon to flow out of the filter screen. A microporous oxygen increasing air stone is placed in the egg collecting barrel, and the air quantity is adjusted to slightly boil. The hatching barrel is a plastic barrel with the bottom radius of 25cm and the height of 1.5m, a pvc pipe with the caliber of 32mm is arranged at the bottom of the barrel, a ball valve assembled at a water outlet is 15cm along with a plastic hose with the caliber of 32mm, and an elbow and the pvc pipe with the caliber of 32mm with the height of 20cm are assembled inside the barrel.

5) And (3) larva washing: every 12 noon, the hatching barrels with the numbers of 1-10 are checked and all hatched normally. The cleaning system device with the number of 11-20 is placed below the hatching barrel. The water in the cleaning system device is clean water treated by a Dow water treater, the water temperature is controlled at 32 ℃, the PH is 8.6, and the total ammonia nitrogen nitrite in the water is 0. The hatching barrel is arranged on a table top 35cm higher than the ground. And (5) turning off the microporous aerator stone in the incubation barrel, and standing for 10 minutes. After 10 minutes, the micropores in the egg collecting barrel are opened to increase oxygen, and the air quantity is adjusted to slightly boil. And opening a pvc ball valve below the hatching barrel, and adjusting the size to ensure that the water flow with the larvae slowly flows into the egg collecting barrel in the corresponding cleaning system device until no water flows out. After collecting the larvae, taking 500ml of water out of the cleaning barrel by using a 500ml beaker, putting 0.08ml of H reagent, uniformly mixing, pouring into the egg collecting barrel, slightly shaking the egg collecting barrel up and down to uniformly distribute the solution in the cleaning barrel and the egg collecting barrel, opening the prepared cleaning water, and slowly flowing the cleaning water into the egg collecting barrel. The cleaning water is clean water treated by a Dow water treater, the water temperature is controlled at 32 ℃, the pH value is 8.4, and the total ammonia nitrogen nitrite in the water is 0. The water in the washing system apparatus was discharged while being introduced, and the total amount of the inflow water was 40 times the volume of the washing tub, and 1600L of the washing water was introduced. After the cleaning, collecting the larvae in the cleaning system device with the number of 11-20, conveying the larvae to an isolated seedling raising pond, and entering a conventional standard seedling raising program.

Example 3

1) Selecting and temporarily culturing parent shrimps: selecting healthy parent shrimps according to the ratio of male to female of 1: 2, wherein the healthy parent shrimps have the body length of 18cm, the body weight of 40g, the body size of the healthy parent shrimps is large, the body color of the healthy parent shrimps is normal, the paraplegia of the healthy parent shrimps is complete, the gill part of the healthy parent shrimps is clean, and no specific virus infection exists. The method for temporarily breeding the parent shrimps comprises the following steps: according to 11 tails/m²The parent shrimps are put into a parent shrimp isolation pond with the size of 25m²The cement pond is 0.6m deep, the isolation pond is soaked in NaOH solution with pH being more than or equal to 14 for 24 hours before use, water in the whole isolation field passes through a Dow water treater, the filter pore diameter is 0.1-0.2 um, water change is carried out according to 40% every day, the water temperature is controlled at 28 ℃ during the whole temporary culture period, the salinity is 28-34 per mill, the dissolved oxygen is ensured to be more than 5mg/L, the pH is 8.7, and the total ammonia nitrogen content in the water is lower than 0.6 mg/L. Feeding parent shrimp feed according to 2% of the weight of the shrimp every day, and feeding for 4 times every day.

2) Infection and enhancement of parent shrimps: after the parent shrimps are temporarily cultured in the isolation area for 10 days, the parent shrimps are fed with the parent shrimp feed mixed with the shrimp liver enterozoon liquid, the mixed feed amount is 10ml/kg, and the feeding amount is 2 percent of the body weight. After 15 days of continuous feeding, fresh excrement produced by parent shrimps in the parent shrimp isolation pool is taken and proved to be positive by PCR detection. And (3) after the detection, removing the eye handle of the female shrimp in the parent shrimp isolation pond every other day, changing the water amount by 60% every day, and controlling the illumination intensity at 500 x. Feeding parent shrimp with feed, sandworm, Loligo chinensis Gray, and hepar bovis Seu Bubali at ratio of 1: 3: 1. The feeding amount is calculated according to 2 percent of the weight of the parent shrimps every day, and the feeding is carried out for 4 times every day. After 20 days of cultivation, the size of the parent shrimps reaches 60 g. The gonads of the female shrimps are full, and the male shrimps are mature.

The preparation method of the shrimp liver intestine cytozoon liquid comprises the following steps: taking excrement from the culture pond with the outbreak of the shrimp liver enterosporidium, the shrimp pond has obvious liver enterosporidium symptom, and PCR detection proves that the shrimp liver enterosporidium is positive. Putting 0.1g of prawn feces of outbreak of shrimp liver enterocytozoon into a homogenizer, quickly adding 1mL of LPBS buffer solution, carrying out bath grinding uniformly, then putting the shrimp liver enterocytozoon liquid into a 1.5mL EP tube, centrifuging for 5min at 3000r/min, taking 200uL of supernatant, adding into 1800uL of PBS buffer solution, and filtering by using a filter which is sterilized at high temperature and is provided with 0.22uL of meshes. 500uL of the filtrate was added to 49.5mL of PBS buffer (pH 7.4), and the mixture was mixed, totaling 50mL, and stored at 4 ℃ until use.

3) Spawning parent shrimps and collecting ova: after the sexual maturity of the parent shrimps is reached, lightly picking out the mature parent shrimps of the gonads of the parent shrimps in the parent shrimp isolation pond at 7 o' clock, checking whether mating is completed, the mated parent shrimps, namely the genitals of the parent shrimps, are firmly clamped by male shrimps and are placed in the spawning isolation pond, the spawning isolation pond is a cement pond with an area of 25 square meters and a water level of 0.6m, the water is clean water treated by a Dow processor, the water temperature is controlled at 29 ℃, the pH is 8.7, the dissolved oxygen is more than 5mg/L, and the total ammonia nitrogen in the water is 0. And (4) putting the unmatched female shrimps back to the net cage of the parent shrimp isolation pond, taking out the net cage after picking up all the female shrimps with full gonads in all the ponds, and returning the un-inseminated female shrimps to the parent shrimp isolation pond. And selecting 11 tails of the female shrimps which are qualified for insemination at night. The length, width and height of the net cage are 1m x 1m, the periphery of the net cage is surrounded by 40 meshes, and the upper surface and the lower surface of the net cage are emptied. When spawning, the device keeps quiet, all lights are turned off, and the spawning rate is improved. And lightly picking out the female shrimps and the parent shrimp isolation pond from the spawning isolation pond at 12 pm. Collecting the eggs in the spawning and leaving pond by using a 300-mesh net cage and a 300-mesh fishing net at 1 point every other morning. The collected parent shrimp eggs were placed in a 20L white bucket. The water in the white barrel is clean water treated by a Dow water treatment device, the water temperature is 29 ℃, the pH value is 8.6, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0. Placing micropore aeration in a white barrel, adjusting air flow to a micro-boiling state, taking 10ml by using a 10ml liquid transfer dropper, putting the 10ml liquid transfer dropper into a 100ml beaker, adding 90ml of clean ultrafiltration water into the beaker, taking 1ml by using a 1ml glass liquid transfer tube, and calculating the tail 13 shrimp egg numbers in the liquid transfer tube, thereby obtaining the total number of the shrimp eggs of 300 ten thousand.

4) And (3) washing eggs: 10 shrimp-containing 500ml water were carefully taken out of the white barrel using a 500ml beaker, and egg collecting barrels of 1 to 10-numbered washing system units were randomly placed, respectively, whereby the number of shrimp eggs in each washing system unit was 75000. The cleaning system device in the cleaning barrel comprises a buckle cleaning barrel and an ovum collecting barrel. The water in the cleaning system device is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the PH is 8.7, the dissolved oxygen is more than 5mg/L, the total ammonia nitrogen nitrite in the water is 0, 2ppm of H solution, namely 0.08ml of H reagent is added into the water, and the mixture is uniformly mixed. The macroscopic mother shrimp feces and impurities are carefully fished out by a 60-mesh fishing net, and the microporous oxygen increasing air stone in the egg collecting barrel is adjusted to be slightly boiled, so that the shrimp eggs are soaked in the solution for 2 minutes. After 2 minutes, the prepared rinsing water was slowly added to the egg collecting bucket. The cleaning water is clean water treated by a Dow water treatment device, the water temperature is controlled at 29 ℃, the pH value is 8.7, and the total ammonia nitrogen nitrite in the water is 0. The water in the washing system apparatus was discharged while being introduced, and the total volume of the inflow water was 20 times the volume of the washing tub, and 800L of the inflow water was counted. After the eggs are washed, the egg collecting barrel is taken out slightly, and the eggs in the egg collecting barrel are transferred to the hatching barrel with the corresponding number of 1-10. The water in the hatching barrel is clean water treated by a Dow water treater, the water level is 1m, the water temperature is 32 ℃, the PH value is 8.4, the dissolved oxygen is more than 5mg/L, and the ammonia nitrogen nitrite in the water is 0. A micropore aeration stone is placed in the hatching barrel, and the air quantity is adjusted to be in a slight boiling state. A50 w incandescent lamp is arranged at a position 40cm above the hatching barrel, and the incandescent lamp is kept in a normally-on state during hatching. The hatching barrel is placed on a table top 35cm away from the ground.

The H reagent is: the anionic polyacrylamide, the povidone iodine and the purified water are prepared according to the proportion of 1: 5: 10. The cleaning system device comprises two parts, wherein the first part is a cleaning barrel, and the cleaning barrel is a square plastic barrel with the length of 40cm, the width of 40cm and the height of 25 cm. The second part is an egg collecting barrel, the radius of the egg collecting barrel is 15cm, the height of the egg collecting barrel is 35cm, a rectangular ring-shaped cutting part is arranged at a position 10cm away from the barrel bottom, the cut holes are 40cm in length and 30cm in width, and a 250-mesh filter screen is fixed in each hole. In the use process, the egg collecting barrel is placed in the cleaning barrel, water is added into the egg collecting barrel, and the water can carry tiny excrement, impurities and shrimp liver enterozoon to flow out of the filter screen. A microporous oxygen increasing air stone is placed in the egg collecting barrel, and the air quantity is adjusted to slightly boil. The hatching barrel is a plastic barrel with the bottom radius of 25cm and the height of 1.5m, a pvc pipe with the caliber of 32mm is arranged at the bottom of the barrel, a ball valve assembled at a water outlet is 15cm along with a plastic hose with the caliber of 32mm, and an elbow and the pvc pipe with the caliber of 32mm with the height of 20cm are assembled inside the barrel.

5) And (3) larva washing: every 12 noon, the hatching barrels with the numbers of 1-10 are checked and all hatched normally. The cleaning system device with the number of 11-20 is placed below the hatching barrel. The water in the cleaning system device is clean water treated by a Dow water treater, the water temperature is controlled at 32 ℃, the PH is 8.7, and the total ammonia nitrogen nitrite in the water is 0. The hatching barrel is arranged on a table top 35cm higher than the ground. And (5) turning off the microporous aerator stone in the incubation barrel, and standing for 10 minutes. After 10 minutes, the micropores in the egg collecting barrel are opened to increase oxygen, and the air quantity is adjusted to slightly boil. And opening a pvc ball valve below the hatching barrel, and adjusting the size to ensure that the water flow with the larvae slowly flows into the egg collecting barrel in the corresponding cleaning system device until no water flows out. After collecting the larvae, taking 500ml of water out of the cleaning barrel by using a 500ml beaker, putting 0.08ml of H reagent, uniformly mixing, pouring into the egg collecting barrel, slightly shaking the egg collecting barrel up and down to uniformly distribute the solution in the cleaning barrel and the egg collecting barrel, opening the prepared cleaning water, and slowly flowing the cleaning water into the egg collecting barrel. The cleaning water is clean water treated by a Dow water treatment device, the water temperature is controlled at 32 ℃, the pH value is 8.7, and the total ammonia nitrogen nitrite in the water is 0. The water in the washing system apparatus was discharged while being introduced, and the total amount of the inflow water was 40 times the volume of the washing tub, and 1600L of the washing water was introduced. After the cleaning, collecting the larvae in the cleaning system device with the number of 11-20, conveying the larvae to an isolated seedling raising pond, and entering a conventional standard seedling raising program.

The above description is only for the specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art can substitute or change the technical solution and the concept of the present invention in the disclosure of the present invention, and fall into the protection scope of the present invention.