A method for screening breeding materials of shrimp anti-vibrio traits

Abstract

The present application relates to the technical field of prawn breeding, and particularly relates to a screening method of prawn Vibrio resistance breeding material. The screening method of prawn Vibrio resistance breeding material comprises the following steps: prawn Vibrio resistance trait stress test, first screening, pathogenic bacteria removal, second screening, breeding and third screening. In the present application, the screened breeding material can be directly used as prawn breeding material, the traits of the breeding material can be accurately and truly fed back, the problems of inaccurate results and the breeding material being unable to be directly used as breeding material in the conventional empirical Vibrio resistance trait test can be avoided, and the precision of prawn breeding material selection and breeding is improved.

Description

Technical Field

[0001] This invention relates to the field of shrimp farming technology, and in particular to a method for screening breeding materials for shrimp with Vibrio resistance traits. Background Technology

[0002] Litopenaeus vannamei, also known as whiteleg shrimp, is a new type of shrimp. However, the existing breeding techniques for new shrimp varieties are generally traditional, and the brand competitiveness is not strong. There is an urgent need to cultivate new shrimp varieties that can meet market demands and have product competitiveness.

[0003] Over the past decade, due to changes in the aquaculture environment, shrimp farming diseases have become more frequent. Among them, vibriosis, represented by early mortality syndrome (EMS), has had a particularly serious impact, and shrimp varieties with strong resistance to vibriosis infection have become a huge demand in the aquaculture market.

[0004] In empirical tests of Vibrio resistance in shrimp, shrimp samples are typically drawn from a population or family for testing. The phenotypic performance of these samples is then used to represent the phenotypic performance of the source population or family. This approach has three significant drawbacks: First, because the tested individuals have been exposed to Vibrio pathogens, they cannot be directly used as breeding material, leading to the elimination of individuals that perform well in the phenotypic test. Second, the phenotypic test results of the samples are used to assess the overall phenotypic characteristics of the population or family, masking the differences between individuals within the population or family. This can result in the inclusion of breeding materials without Vibrio resistance in subsequent parent selection, potentially leading to the failure of the Vibrio resistance breeding program. Third, shrimp with excellent Vibrio resistance traits may be individuals with genetic variations or unique genotypes within the population or family, and their phenotypic characteristics may not be representative of the entire population or family. Therefore, the screening method of using individual samples to represent the Vibrio resistance traits of a family or population loses its accuracy.

[0005] Therefore, it is necessary to provide a new method for screening breeding materials with Vibrio resistance in shrimp to address the above-mentioned shortcomings. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention provides a method for screening breeding materials for Vibrio resistance in shrimp. The selected materials can be directly used as shrimp breeding materials, providing accurate and reliable feedback on their traits. This avoids the significant problems of inaccurate results and the inability to directly use test subjects as breeding materials in conventional empirical Vibrio resistance tests, thereby improving the accuracy of shrimp breeding material selection.

[0007] The purpose of this invention is to provide a method for screening breeding materials for shrimp with Vibrio resistance traits.

[0008] The method for screening breeding materials for Vibrio resistance in shrimp according to a specific embodiment of the present invention includes the following steps:

[0009] (1) Stress test of anti-Vibrio trait in shrimp

[0010] After temporarily raising the shrimp larvae in the test tank for one week, the seawater in the test tank was drained to 30-40%, and a bacterial solution containing pathogens was added to the test tank for 12-16 hours of immersion infection. Then, the remaining 50% of the seawater in the test tank was drained, and clean seawater was added to the water level of the temporary holding process, and the shrimp were continued to be raised.

[0011] (2) First screening

[0012] After the immersion infection, observe the activity and feeding of shrimp in the test pond 3 to 5 times a day, remove dead or dying shrimp, continue for 2 to 5 days, and retain the surviving shrimp after one screening for continued feeding.

[0013] (3) Elimination of pathogens

[0014] After one screening is completed, a pathogen removal process is performed, including:

[0015] The test pool is changed daily and disinfected every three days.

[0016] Medicated feed should be administered 0–7 days after the pathogenic bacteria have been eliminated.

[0017] 7–21 days after the pathogenic bacteria are eliminated, discontinue medicated feed and administer probiotic-containing feed.

[0018] (4) Secondary screening

[0019] In the process of eliminating pathogens in shrimp farming, the test ponds should be checked every morning, noon and evening. Dead or dying shrimp should be removed from the test ponds, and surviving shrimp should be kept.

[0020] (5) Aquaculture

[0021] After pathogen removal and secondary screening, the surviving shrimp continue to be cultured for 70–90 days;

[0022] (6) Three-stage screening

[0023] Collect feces from farmed shrimp, test for pathogens in the feces, select shrimp with negative test results, and complete the screening of breeding materials for shrimp anti-Vibrio trait.

[0024] Prepare multiple buckets, distribute the cultured shrimp into the buckets, feed them, collect shrimp feces from each bucket, and combine the shrimp that test negative by PCR amplification to complete the screening of breeding materials for shrimp anti-Vibrio trait.

[0025] According to the specific embodiments of the present invention, in the method for screening breeding materials for shrimp with Vibrio resistance traits, in step (1) of the stress test for shrimp with Vibrio resistance traits, the density of shrimp raised in the test pond is 300-1500 shrimp / m³.3 The body length of the prawn is 2-3 cm.

[0026] According to the specific embodiments of the present invention, in the method for screening breeding materials for shrimp with anti-Vibrio trait, in step (1) the stress test of shrimp anti-Vibrio trait, the pathogenic bacteria is Vibrio parahaemolyticus or Vibrio harzianum; the OD of the pathogenic bacteria solution 600nm ≥2.0.

[0027] According to the specific embodiments of the present invention, in the method for screening breeding materials for Vibrio resistance in shrimp, in step (1) of the stress test for Vibrio resistance in shrimp, during the immersion infection process, the concentration of pathogenic bacteria in the test tank is ≥1×10⁻⁶. 5 cfu / mL.

[0028] According to the method for screening breeding materials for anti-Vibrio trait in shrimp provided by a specific embodiment of the present invention, in step (3) pathogen removal, the water exchange volume is as follows:

[0029] The water exchange rate is 20-30% from 0 to 7 days.

[0030] The water exchange rate is 10-20% every 7-21 days.

[0031] According to the specific embodiments of the present invention, in the method for screening breeding materials for shrimp with anti-Vibrio trait, in step (3) pathogen removal, the sterilization is carried out by disinfecting the water with chlorine dioxide.

[0032] According to the specific embodiments of the present invention, in the method for screening breeding materials for shrimp with anti-Vibrio trait, in step (3) pathogen removal, the medicated feed contains 1-5‰ florfenicol.

[0033] According to the specific embodiments of the present invention, the method for screening breeding materials for shrimp with anti-Vibrio trait is provided in step (3) pathogen removal, wherein the feed contains 0.5-1% Bacillus pumilus.

[0034] According to the specific embodiments of the present invention, in the method for screening breeding materials for anti-Vibrio trait in shrimp, the capacity of the water bucket in the three screening steps (6) is 20-30L.

[0035] According to the specific embodiments of the present invention, in the method for screening breeding materials for anti-Vibrio trait in shrimp, in step (6) three screenings, the number of shrimp in the water bucket is 10 per bucket.

[0036] Compared with the prior art, the beneficial effects of the present invention are:

[0037] 1. After screening, the material in this invention can be directly used as a breeding material for shrimp, which can accurately reflect the characteristics of the breeding material and avoid the prominent problems of inaccurate results and the inability of test subjects to be directly used as breeding materials in conventional empirical anti-Vibrio trait tests, thereby improving the accuracy of shrimp breeding material selection.

[0038] 2. The screening method of the present invention can avoid the prominent problems of inaccurate results and the inability of test subjects to be directly used as breeding materials in conventional empirical anti-Vibrio trait tests, thereby improving the accuracy of shrimp breeding material selection.

[0039] 3. This invention uses bacterial solutions containing pathogenic bacteria to screen shrimp, which can obtain breeding materials with higher breeding value. Since the enhanced resistance to Vibrio in shrimp is not an inherent characteristic of a family or population, through large-scale Vibrio infection testing and a high-ratio strict culling, shrimp individuals with significant resistance to Vibrio can be screened out. These individuals are a few individuals in the family or population with unique genotypes or acute variants. At the same time, during the screening process, a large number of shrimp infected with pathogenic bacteria are removed, especially shrimp with latent pathogenic bacteria infection, thus reducing the harm caused by pathogenic bacteria by not carrying pathogenic bacteria.

[0040] 4. In the screening method of this invention, the concentration of pathogenic Vibrio is diluted by frequent water changes, chlorine dioxide is used to kill pathogenic Vibrio residual in the water environment, chlorfenapyr, which has a broad-spectrum and highly effective effect against Vibrio and is permitted for use, is used to inhibit the growth of Vibrio in shrimp, and during the shrimp's skill recovery period, Bacillus subtilis, which has a broad-spectrum antagonistic effect against Vibrio, is used to inhibit Vibrio in shrimp, thus promptly removing infected shrimp and reducing the probability of pathogen transmission. In this way, the introduced pathogens are completely removed from shrimp and the aquaculture environment through multiple pathways. Furthermore, sensitive PCR detection is used to ensure that the screened shrimp breeding stock does not carry EMS pathogens, thus ensuring the quality of the breeding materials.

[0041] 5. The shrimp breeding materials screened using the method of this invention have strong survival adaptability and strong resistance to Vibrio EMS. Detailed Implementation

[0042] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0043] Example 1

[0044] This embodiment provides a method for screening breeding materials for Vibrio resistance in shrimp, including the following steps:

[0045] 1. Culture of pathogenic bacteria

[0046] (1) Vibrio parahaemolyticus SC1907 was used as the pathogenic strain causing EMS;

[0047] (2) Take Vibrio parahaemolyticus SC1907 and culture it overnight in LB medium. Culture conditions: 30℃, shaking speed 200 rpm, culture time 24 h, until OD. 600nm ≥3.0;

[0048] (3) Add LB medium for quantitative dilution to obtain bacterial solution containing pathogenic bacteria.

[0049] 2. Stress test of anti-Vibrio trait in shrimp

[0050] (1) Select a group or family of Litopenaeus vannamei as test shrimp, with 10,000 individuals and an average body length of 2.5 cm.

[0051] (2) In an area of ​​10m 2 10m of clean seawater was added to the test tank. 3 The test object is transferred to the test pool and temporarily held for 16 hours;

[0052] (3) Drain the water in the test tank to a depth of 0.35m, add bacterial solution to the test tank, and make the concentration of pathogenic bacteria in the test tank water 1×10⁻⁶. 5 cfu / mL, begin immersion in the immersion bath for infection;

[0053] (4) After the shrimp were immersed in the infection for 14 hours, 50% of the water in the test tank was drained, and clean seawater was added back to the test tank until the water depth was 1m. The shrimp were then fed according to the conventional feeding and management methods.

[0054] 3. First screening

[0055] Within 72 hours of immersion infection, the activity and feeding of shrimp in the test pond were observed four times a day. Dead and dying shrimp were removed in a timely manner, and the remaining shrimp were selected as breeding materials through sequential screening.

[0056] 4. Elimination of pathogenic Vibrio and secondary screening

[0057] (1) Mix florfenicol (purchased from Shandong Dexing Biotechnology Co., Ltd.) into the conventional shrimp feed at a mass percentage of 2‰, and add commonly used adhesives (purchased from Yuncheng Aowei Veterinary Drug Co., Ltd.). After drying, prepare the medicated feed and store it in a refrigerator at 4℃ for later use.

[0058] (2) Mix Bacillus pumilus (purchased from Guangzhou Nuojing Biotechnology Co., Ltd.) into conventional shrimp feed at a mass percentage of 0.75%, and add commonly used adhesives (purchased from Yuncheng Aowei Veterinary Drug Co., Ltd.). After drying, prepare the bacterial feed and store it in a refrigerator at 4°C for later use.

[0059] (3) Feed the medicated feed according to the normal feed feeding method. 1.5 hours after feeding, observe the activity of the shrimp in the test pond and remove the shrimp that are not moving normally, or have empty intestines or empty stomachs. Check the test pond three times a day, in the morning, noon and evening, and remove dead or dying shrimp in time. During the feeding period, change the water in the test pond every day, and the water change volume is 25%.

[0060] (4) After feeding the medicated feed for 7 days, stop feeding the medicated feed and start feeding the bacterial feed according to the normal feed feeding method. Feed the bacterial feed continuously for 14 days. Check the test pond three times a day, in the morning, noon and evening, and remove dead or dying shrimp in time. During the feeding of bacterial feed, change the water in the test pond every day, with a water change volume of 15%.

[0061] (5) During the feeding of medicated feed and bacterial feed, the water in the test pool is disinfected every 72 hours. Chlorine dioxide is used for disinfection. 12 hours after disinfection, 50% of the pool water in the test pool is drained and then clean seawater is added to the normal water level.

[0062] 5. Normal breeding

[0063] After pathogenic Vibrio removal and secondary screening are completed, the animals enter the normal indoor rearing process at a stocking density of 120 fish / m². 3 Indoor cultivation for 80 days.

[0064] 6. Three rounds of screening

[0065] (1) After 80 days of normal culture, feed the shrimp with conventional feed. One hour after feeding, test each shrimp individually and select individuals with intact body shape, normal size and color of hepatopancreas, clear gastrointestinal boundaries, and full gastrointestinal contents.

[0066] (2) Distribute the shrimp individuals into 25L water buckets at a rate of 10 shrimp per bucket, and continue to feed them according to the conventional farming method. After feeding for 3 hours, collect the shrimp feces in each bucket with a pipette, put them into 1.5mL centrifuge tubes, centrifuge for 2 minutes, and absorb excess water.

[0067] (3) DNA was extracted using a bacterial genome extraction kit (TaKaKa) and the EMS pathogens in the feces were detected by PCR for the first time (see reference 1 for the PCR detection method) to obtain negative or positive results.

[0068] (4) Continue feeding the shrimp in the bucket until the third day, collect shrimp feces from the bucket again, and obtain negative or positive test results according to the aforementioned test method;

[0069] (5) The shrimp that tested negative twice were merged into one pond, with a total of 210 shrimp retained, a retention rate of 2.1%. The 210 retained shrimp were used as the final breeding material and entered the next stage of broodstock breeding.

[0070] The breeding materials in this embodiment have the following characteristics:

[0071] Thirty breeding materials were randomly selected from the above and temporarily stored in a test tank containing 200L of clean seawater for 16 hours. A bacterial solution of Vibrio parahaemolyticus SC1907 was added to the test tank at a concentration of 1×10⁻⁶. 5 The cfu / mL concentration was used for infection testing. After 14 hours of infection testing, 80% of the seawater in the test tank was drained, and clean seawater was added back to bring the total seawater volume in the test tank to 200L. The shrimp were then fed according to conventional feeding and management methods. At the start of the infection test, the activity and feeding of the shrimp in the test tank were observed four times a day. Dead or dying shrimp were removed in a timely manner until the number of survivors was counted after 7 days.

[0072] Experimental results: 28 fish survived, with a survival rate of 93.3%.

[0073] Example 2

[0074] This embodiment provides a method for screening breeding materials for Vibrio resistance in shrimp, including the following steps:

[0075] 1. Culture of pathogenic bacteria

[0076] (1) Vibrio harveyi was used as the pathogenic Vibrio strain that causes EMS;

[0077] (2) Take Vibrio harveyi and culture it overnight in LB medium. Culture conditions: 30℃, shaking speed 200 rpm, culture time 24 h, until OD. 600nm ≥3.0;

[0078] (3) Add LB medium for quantitative dilution to obtain a bacterial suspension containing pathogenic bacteria.

[0079] 2. Stress test of anti-Vibrio trait in shrimp

[0080] (1) Select a group or family of Litopenaeus vannamei as test shrimp, with 10,000 individuals and an average body length of 2.5 cm.

[0081] (2) In an area of ​​10m 2 10m of clean seawater was added to the test tank. 3The test object is transferred to the test pool and temporarily held for 16 hours;

[0082] (3) Drain the water in the test tank to a depth of 0.35m, add bacterial solution to the test tank, and make the concentration of pathogenic bacteria in the test tank water 1×10⁻⁶. 5 cfu / mL, begin immersion in the immersion bath for infection;

[0083] (4) After the shrimp were immersed in the infection for 14 hours, 50% of the water in the test tank was drained, and clean seawater was added back to the test tank until the water depth was 1m. The shrimp were then fed according to the conventional feeding and management methods.

[0084] 3. First screening

[0085] Within 72 hours of immersion infection, the activity and feeding of shrimp in the test pond were observed four times a day. Dead and dying shrimp were removed in a timely manner, and the remaining shrimp were selected as breeding materials through sequential screening.

[0086] 4. Elimination of pathogenic Vibrio and secondary screening

[0087] (1) Mix florfenicol (Shandong Dexing Biotechnology Co., Ltd.) into the conventional shrimp feed at a mass percentage of 2‰, add commonly used adhesives, dry and then make medicated feed. Store in a refrigerator at 4℃ for later use.

[0088] (2) Mix Bacillus pumilus (Guangzhou Nuojing Biotechnology Co., Ltd.) into the feed of urban shrimp at a mass percentage of 0.75%, add commonly used adhesives, dry and make bacterial feed, and store in a refrigerator at 4℃ for later use.

[0089] (3) Feed the medicated feed according to the normal feeding method. 1.5 hours after feeding, observe the condition of the shrimp in the test pond and remove the shrimp that are not moving normally, have empty intestines or empty stomachs. Check the test pond three times a day, morning, noon and evening, and remove dead or dying shrimp in time. During the feeding period, change the water in the test pond every day, and the water change volume is 25%.

[0090] (4) After feeding the medicated feed for 7 days, stop feeding the medicated feed and start feeding the bacterial feed according to the normal feed feeding method. Feed the bacterial feed continuously for 14 days. Check the test pond three times a day, morning, noon and evening, and remove dead or dying shrimp in time. During the feeding of bacterial feed, change the water in the test pond every day, with a water change volume of 20%.

[0091] (5) During the feeding of medicated feed and bacterial feed, the water in the test pool is disinfected every 72 hours. Chlorine dioxide is used for disinfection. 12 hours after disinfection, 50% of the pool water in the test pool is drained and then clean seawater is added to the normal water level.

[0092] 5. Normal breeding

[0093] After pathogenic Vibrio removal and secondary screening are completed, the animals enter the normal indoor rearing process at a stocking density of 120 fish / m². 3 Indoor cultivation for 80 days.

[0094] 6. Three rounds of screening

[0095] (1) After 80 days of normal culture, feed the shrimp with conventional feed. One hour after feeding, test each shrimp individually and select individuals with intact body shape, normal size and color of hepatopancreas, clear gastrointestinal boundaries, and full gastrointestinal contents.

[0096] (2) Distribute the shrimp individuals into 25L water buckets at a rate of 10 shrimp per bucket, and continue to feed them according to the conventional farming method. After feeding for 3 hours, collect the shrimp feces in each bucket with a pipette, put them into 1.5mL centrifuge tubes, centrifuge for 2 minutes, and absorb excess water.

[0097] (3) DNA was extracted using a bacterial genome extraction kit (TaKaKa) and the EMS pathogens in the feces were detected by PCR for the first time to obtain negative or positive results;

[0098] (4) Continue feeding the shrimp in the bucket until the third day, collect shrimp feces from the bucket again, and obtain negative or positive test results according to the aforementioned test method;

[0099] (5) The shrimp that tested negative twice were merged into one pond, with a total of 250 shrimp retained, a retention rate of 2.5%. The 250 retained shrimp were used as the final breeding material and entered the next stage of broodstock breeding.

[0100] The breeding materials in this embodiment have the following characteristics:

[0101] Thirty breeding materials were randomly selected from the above and temporarily stored in a test tank containing 200L of clean seawater for 16 hours. A bacterial solution of Vibrio parahaemolyticus SC1907 was added to the test tank at a concentration of 1×10⁻⁶. 5 The cfu / mL concentration was used for infection testing. After 14 hours of infection testing, 80% of the seawater in the test tank was drained, and clean seawater was added back to bring the total seawater volume in the test tank to 200L. The shrimp were then fed according to conventional feeding and management methods. At the start of the infection test, the activity and feeding of the shrimp in the test tank were observed four times a day. Dead or dying shrimp were removed in a timely manner until the number of survivors was counted after 7 days.

[0102] Experimental results: 26 fish survived, with a survival rate of 86.7%.

[0103] Comparative Example

[0104] This comparative example is based on empirical testing of shrimp anti-Vibrio trait. Thirty shrimp from the EMS-resistant family identified by conventional anti-Vibrio trait detection methods were randomly selected as control materials for the following comparative experiment:

[0105] The control material was temporarily stored in a test tank containing 200L of clean seawater and incubated for 16 hours; a bacterial suspension of Vibrio parahaemolyticus SC1907 was added to the test tank at a concentration of 1×10⁻⁶. 5 The cfu / mL concentration was used for infection testing. After 14 hours of infection testing, 80% of the seawater in the test tank was drained, and clean seawater was added back to bring the total seawater volume in the test tank to 200L. The shrimp were then fed according to conventional feeding and management methods. At the start of the infection test, the activity and feeding of the shrimp in the test tank were observed four times a day. Dead or dying shrimp were removed in a timely manner until the number of survivors was counted after 7 days.

[0106] Comparative experiment results: 20 fish survived, with a survival rate of 66.7%.

[0107] Reference file 1: Han JE, Tang KF, Tran LH, Lightner DV. Photorhabdus insect-related(Pir)toxin-like genes in a plasmid of Vibrio parahaemolyticus, thecausative agent of acute hepatopancreatic necrosis disease(AHPND)ofshrimp.Diseases of Aquatic Organisms.2015,113(1):33-40.

[0108] 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 principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for screening breeding materials for vibrio resistance in shrimp, characterized in that, Includes the following steps: (1) Stress test of anti-Vibrio trait in shrimp After temporarily raising the shrimp larvae in the test for one week, the seawater in the test tank was drained to 30-40%, and a bacterial solution containing pathogens was added to the test tank for 12-16 hours of immersion infection. Then, the remaining 50% of the seawater in the test tank was drained, and clean seawater was added to the water level of the temporary raising process, and the shrimp were continued to be raised. (2) First screening After the immersion infection, observe the activity and feeding of shrimp in the test pond 3 to 5 times a day, remove dead or dying shrimp, continue for 2 to 5 days, and retain the surviving shrimp after one screening for continued feeding. (3) Elimination of pathogens After one screening is completed, a pathogen removal process is performed, including: The test pool is changed daily and disinfected every three days. Medicated feed should be administered 0–7 days after the pathogenic bacteria have been eliminated. 7–21 days after the pathogenic bacteria are eliminated, discontinue medicated feed and administer probiotic-containing feed. (4) Secondary screening During the process of eliminating pathogens, the test ponds are checked every morning, noon and evening. Dead or dying shrimp are removed from the test ponds, while surviving shrimp are kept. (5) Aquaculture After pathogen removal and secondary screening, the surviving shrimp continue to be cultured for 70–90 days; (6) Three screenings Collect feces from farmed shrimp, test for pathogens in the feces, select shrimp with negative test results, and complete the screening of breeding materials for shrimp anti-Vibrio trait. Prepare multiple buckets, distribute the cultured shrimp into the buckets, feed them, collect shrimp feces from each bucket, and combine the shrimp that test negative by PCR amplification to complete the screening of breeding materials for shrimp anti-Vibrio trait.

2. The method for screening breeding materials for Vibrio resistance in shrimp according to claim 1, characterized in that, In step (1), the shrimp density in the test pond for Vibrio resistance was 300–1500 shrimp / m³. 3 The body length of the prawn is 2-3 cm.

3. The method for screening breeding materials for Vibrio resistance in shrimp according to claim 1, characterized in that, In step (1), during the stress test of shrimp's anti-Vibrio trait, the pathogenic bacteria are Vibrio parahaemolyticus or Vibrio harzianum; the OD of the pathogenic bacteria solution... 600nm ≥2.

0.

4. The method for screening breeding materials for shrimp with Vibrio resistance according to claim 1, characterized in that, In step (1), during the stress test of shrimp's anti-Vibrio trait, the concentration of pathogenic bacteria in the test tank during the immersion infection process was ≥1×10⁻⁶. 5 cfu / mL.

5. The method for screening breeding materials for Vibrio resistance in shrimp according to claim 1, characterized in that, In step (3) of pathogen removal, the amount of water replaced is: The water exchange rate for 0–7 days is 20–30%. The water exchange rate is 10-20% every 7-21 days.

6. The method for screening breeding materials for vibrio resistance in shrimp according to claim 1, characterized in that, In step (3) pathogen removal, chlorine dioxide is used to disinfect the water.

7. The method for screening breeding materials for vibrio resistance in shrimp according to claim 1, characterized in that, In step (3) pathogen elimination, the medicated feed contains 1-5‰ florfenicol.

8. The method for screening breeding materials for vibrio resistance in shrimp according to claim 1, characterized in that, In step (3) pathogen removal, the feed contains 0.5-1% of Bacillus simulans.

9. The method for screening breeding materials for vibrio resistance in shrimp according to claim 1, characterized in that, In step (6), during the three screenings, the capacity of the water bucket is 20-30L.

10. The method for screening breeding materials for vibrio resistance in shrimp according to claim 1, characterized in that, In step (6), during the three screenings, the number of shrimp in the bucket is 10 per bucket.