CN105746410A - Production method of cryptocaryon irritus-resistant epinephelus fuscoguttatus and epinephelus lanceolatus fry - Google Patents
Production method of cryptocaryon irritus-resistant epinephelus fuscoguttatus and epinephelus lanceolatus fry Download PDFInfo
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- 238000004519 manufacturing process Methods 0.000 title claims abstract description 15
- 241001217936 Epinephelus lanceolatus Species 0.000 title abstract description 7
- 241001149251 Epinephelus fuscoguttatus Species 0.000 title abstract description 6
- ORIGEOXWTMPZQD-DUFGSWQCSA-N Cryptocaryon Natural products O[C@H]1C=C[C@H]2OC(=O)C[C@H]2[C@@H]1C(=O)C=Cc3ccccc3 ORIGEOXWTMPZQD-DUFGSWQCSA-N 0.000 title abstract 6
- 241001663425 Cryptocaryon Species 0.000 title abstract 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 32
- 208000015181 infectious disease Diseases 0.000 claims abstract description 24
- 230000036039 immunity Effects 0.000 claims abstract description 21
- 241000238631 Hexapoda Species 0.000 claims description 55
- 241000251468 Actinopterygii Species 0.000 claims description 51
- 238000000034 method Methods 0.000 claims description 36
- 241001417495 Serranidae Species 0.000 claims description 32
- 239000013535 sea water Substances 0.000 claims description 23
- 231100000614 poison Toxicity 0.000 claims description 18
- 230000004936 stimulating effect Effects 0.000 claims description 18
- 239000003440 toxic substance Substances 0.000 claims description 18
- 230000000638 stimulation Effects 0.000 claims description 17
- 241000321428 Epinephelus guttatus Species 0.000 claims description 16
- 230000007794 irritation Effects 0.000 claims description 14
- 241001663423 Cryptocaryon irritans Species 0.000 claims description 11
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 11
- 238000005538 encapsulation Methods 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- 238000009395 breeding Methods 0.000 claims description 8
- 230000001488 breeding effect Effects 0.000 claims description 8
- 230000002265 prevention Effects 0.000 claims description 7
- 238000007789 sealing Methods 0.000 claims description 7
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims description 6
- 208000003322 Coinfection Diseases 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 230000000694 effects Effects 0.000 claims description 6
- 239000013505 freshwater Substances 0.000 claims description 6
- 230000001524 infective effect Effects 0.000 claims description 6
- 229910052740 iodine Inorganic materials 0.000 claims description 6
- 239000011630 iodine Substances 0.000 claims description 6
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 claims description 6
- 241000630524 Taractes rubescens Species 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 5
- 238000012360 testing method Methods 0.000 claims description 5
- 229930182555 Penicillin Natural products 0.000 claims description 4
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 4
- 229940049954 penicillin Drugs 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 230000012447 hatching Effects 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 230000001681 protective effect Effects 0.000 claims description 3
- 229960005322 streptomycin Drugs 0.000 claims description 3
- 238000005259 measurement Methods 0.000 claims 1
- 201000010099 disease Diseases 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 5
- 230000001900 immune effect Effects 0.000 abstract description 2
- 230000006378 damage Effects 0.000 description 5
- 230000008901 benefit Effects 0.000 description 4
- 208000035240 Disease Resistance Diseases 0.000 description 2
- 241000357444 Epinephelus coioides Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 208000030852 Parasitic disease Diseases 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- KKAJSJJFBSOMGS-UHFFFAOYSA-N 3,6-diamino-10-methylacridinium chloride Chemical compound [Cl-].C1=C(N)C=C2[N+](C)=C(C=C(N)C=C3)C3=CC2=C1 KKAJSJJFBSOMGS-UHFFFAOYSA-N 0.000 description 1
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 description 1
- 241000357439 Epinephelus Species 0.000 description 1
- 241000692870 Inachis io Species 0.000 description 1
- 229960000266 acriflavinium chloride Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- PLHJDBGFXBMTGZ-WEVVVXLNSA-N furazolidone Chemical compound O1C([N+](=O)[O-])=CC=C1\C=N\N1C(=O)OCC1 PLHJDBGFXBMTGZ-WEVVVXLNSA-N 0.000 description 1
- 229960001625 furazolidone Drugs 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 238000009364 mariculture Methods 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 1
- 229960000282 metronidazole Drugs 0.000 description 1
- 230000024241 parasitism Effects 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 229960001841 potassium permanganate Drugs 0.000 description 1
- 229960002668 sodium chloride Drugs 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- JNMRHUJNCSQMMB-UHFFFAOYSA-N sulfathiazole Chemical compound C1=CC(N)=CC=C1S(=O)(=O)NC1=NC=CS1 JNMRHUJNCSQMMB-UHFFFAOYSA-N 0.000 description 1
- 229960001544 sulfathiazole Drugs 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Fodder In General (AREA)
Abstract
The invention relates to a production method of cryptocaryon irritus-resistant epinephelus fuscoguttatus and epinephelus lanceolatus fry, belongs to the field of disease-resistant fry production and in particular belongs to the field of cryptocaryon irritus-resistant fry production. The production method firstly adopts a mode of temperature-varying infection and temperature-varying cultivation to produce disease-resistant epinephelus fuscoguttatus and epinephelus lanceolatus fry, and a certain temperature difference is kept between the water temperature during infection and the cultivation water temperature within 10 days after infection, so that the immune effect of living larvae of cryptocaryon irritus can be enhanced, rotational cultivation can be combined, the cryptocaryon irritus-resistant epinephelus fuscoguttatus and epinephelus lanceolatus fry with a longer effective immune protection period is obtained, and the cryptocaryon irritus-resistant epinephelus fuscoguttatus and epinephelus lanceolatus fry still has 58 percent of relative protection rate in the eighth month after immunity and 32 percent of relative protection rate in the ninth month.
Description
Technical field
This technology relates to the production method of a kind of irritation cryptonucleus insect Margarita Radix Gentianae grouper fry, belongs to disease resistance fry production field, is specially irritation cryptonucleus insect fry production field.
Background technology
Margarita Radix Gentianae cabrilla (Epinephelusfuscoguttatus×Epinepheluslanceolatus), for the novel cabrilla that section, the female brown some cabrilla of Epinephelus and the hybridization of male epinephelus lanceolatus fish are cultivated, it has male parent and maternal advantage concurrently, fast growth, strong stress resistance, and delicious meat, become the new lover of fishes of mariculture in China, have cultivating large area on the ground such as Guangdong and Hainan.
Stimulate cryptocaryoniosis to be commonly called as marine fish ichthyophthiriasis, be by a kind of seawater fish obligatory parasitism ciliate---the lethal parasitic disease (ColorniandBurgess, 1997) that-stimulation cryptonucleus insect (Cryptocaryonirritans) causes.In recent years, cryptocaryoniosis is stimulated to have become as one of the most serious parasitic disease of China's harm, huge harm (Luoetal. is caused to sea water fishery, 2008), it is listed in country's " one, two, three class animal epidemic disease kind registers " unique aquatic animal two class epidemic disease (Ministry of Agriculture, 2008).The host range of stimulation cryptonucleus insect is very extensive, and except stimulation cryptonucleus insect being had stronger resistance except cartilaginous fish, Marine teleost class generally can be stimulated cryptonucleus insect and infect, and wherein Margarita Radix Gentianae cabrilla is also one of object stimulating cryptonucleus insect infection.
Owing to stimulating the complexity of cryptonucleus insect life cycle so that the prevention and control of this disease are extremely difficult.Production at present upper main employing medical treatment such as formalin, copper sulfate, acriflavinium chloride, peacock green, methylene blue, quinoline woods group, metronidazole, potassium permanganate, sodium chloride, sulfathiazole, furazolidone and penicillin etc. (ColorniandBurgess, 1997；Cai Ze equality, 2001；Noritakaetal., 2001；Noritakaetal., 2003；Huang Wei etc., 2004), but these methods are owing to depositing many defects, such as effect instability, destroy breeding environment, harm food safety, enforcement inconvenience etc., are extremely difficult to desirable effect.
Colorni (1987) proposes to stimulate cryptocaryoniosis with different physical method preventing and treatings, and one of which method is that fish carries out rotation raising in 2 aquariums, and vacant aquarium is dried and cleaning processes.This method is very effective, but he does not provide concrete operational approach, in particular without the frequency providing rotation under different temperatures and the time the two critical data altogether needed.Zhang Tuanwei (2012) determines avette silvery pomfret and Epinephelus coioides infects the rotation frequency stimulating cryptonucleus insect and rotational cycle at different temperatures, but does not measure whether this method can bring out marine fish generation acquired immunity protection.
Study confirmation at present, seawater fish to stimulate cryptonucleus insect infection can produce acquired immunity protection, vaccination be likely to become preventing and treating stimulate cryptocaryoniosis effective ways (Yoshinagaetal, 1997；Yambotetal, 2006；Luoetal, 2007；HatanakaandHirazawa, 2008).Recent study person has carried out some basic research stimulating cryptonucleus insect immune protection and applied researcies; discovery cryptocaryon irritans larva infects marine fish such as Epinephelus coioides, avette silvery pomfret etc. through body surface; can reach more than 80% immune protective rate (Luoetal, 2007；But learn bright etc., 2008；Yoshinagaetal, 1997), but effective immune period of these experiments is the longest be can only achieve 3 months, and the not research different temperatures impact on marine fish acquired immunity protection in detail.
In actual production process; in the urgent need to the Margarita Radix Gentianae grouper fry that a kind of effective immune period is longer; for meeting this needs, applicant, through test of many times and trial, has found out a kind of effective immune period production method longer, irritation cryptonucleus insect Margarita Radix Gentianae grouper fry.
Summary of the invention
It is contemplated that overcome, the Margarita Radix Gentianae grouper fry immune period of irritation cryptonucleus insect is too short, be not suitable with the shortcomings such as industrialized aquaculture production, it is provided that the production method of the irritation cryptonucleus insect Margarita Radix Gentianae grouper fry that a kind of practicality is stronger.
The present invention infects adopting unique alternating temperature and rotation cultural method; cultivate the Margarita Radix Gentianae grouper fry that irritation cryptonucleus insect infects; the immune protective efficiency stimulating cryptonucleus insect to be reached more than 8 months by this Margarita Radix Gentianae lithosporic fry; the relative protection ratio that after immunity, 8th month also has 58%, the relative protection ratio still having 32% on the 9th month.
The present invention is unique in that; the mode that alternating temperature infects and alternating temperature cultivates is adopted to produce the grouper fry of disease resistance first; water temperature during infection keeps a fixed difference difference with the temperature of cultivation infected in latter 10 days; thus strengthening the immune effect stimulating cryptonucleus insect larva alive; and then combine with rotation cultivation, it is thus achieved that the irritation cryptonucleus insect Margarita Radix Gentianae grouper fry that effectively immune period is longer.
Advantage of the invention is that; solve the technical problem that the seawater fish seed immune period always perplexing raiser is short; adopt Promethean infection breeding way; produce the irritation cryptonucleus insect Margarita Radix Gentianae grouper fry that effective immune period was up to 8 months; it is applicable to the Margarita Radix Gentianae cabrilla healthy aquaculture of cultivation factory and raiser; the massive losses that sea water ichthyophthiriasis brings can be reduced or remitted, be greatly improved the culture benefit of fisherman.
The present invention is achieved by the following technical solutions:
1. the collection and the concentration that stimulate cryptonucleus insect larva alive calculate
Utilize special stimulation cryptonucleus insect legacy system to stimulating cryptonucleus insect go down to posterity and collect, its host of going down to posterity is for avette silvery pomfret, gather well-developed stimulation cryptonucleus insect encapsulation, be placed in sterilized clean beaker and hatch, Spawning water be autoclaved, add the nature seawater of appropriate penicillin and streptomycin；Collecting and hatch 2 hours interior cryptocaryon irritans larva liquid, fully take 50 μ L worm liquid after mixing, add 10 μ L formaldehyde and killed by larva, count under the microscope, note calculates the worm concentration alive of larva liquid.
2. Margarita Radix Gentianae grouper fry is raised and train, is infected
Purchase the Margarita Radix Gentianae grouper fry of about 13g, infect first 2 weeks and fry is carried out domesticating of examination, until its feed, activity are normally, in whole breeding process, raising the use water strainer filtering of 600 orders, its salinity is about 30 ‰, pH is about 8, and dissolved oxygen amount is at more than 5.6mg/L；Margarita Radix Gentianae grouper fry is divided into matched group and experimental group (1-9 group) totally ten groups, experimental group is often organized fry and is infected with hatching 2 hours interior cryptocaryon irritans larvas, during infection water temperature 1-3 group be 24 DEG C, 4-6 group be 27 DEG C, 7-9 group be 30 DEG C, infecting water consumption is 1L sea water/tail fish, and infective dose is 8000 larvas/tail fish；Respectively fry being put in different alternate culture ponds after infecting 6 hours, wherein the 1st, 4,7 groups of water temperatures control at 24 DEG C, and the 2nd, 5,8 groups of water temperatures control at 27 DEG C, and the 3rd, 6,9 groups of water temperatures control at 30 DEG C；Matched group does not carry out stimulating cryptonucleus insect to infect, and temperature of cultivation is 27 DEG C.Alternate culture pond length 40 meters, wide 1.5 meters, high 1 meter, centre, pond seals dividing plate and is divided into two by culturing pool, ensureing both sides water not intercommunication, metainfective fry is totally placed in the side in pond, adds betagen solution (available iodine content is 10%) the prevention secondary infection of final concentration of 2ppm.Concrete packet situation is shown in following table:
| Group number | Infect water temperature (DEG C) | Temperature of cultivation (DEG C) |
| Matched group | It is uninfected by | 27 |
| 1 | 24 | 24 |
| 2 | 24 | 27 2 --> |
| 3 | 24 | 30 |
| 4 | 27 | 24 |
| 5 | 27 | 27 |
| 6 | 27 | 30 |
| 7 | 30 | 24 |
| 8 | 30 | 27 |
| 9 | 30 | 30 |
3, the rotation cultivation of Margarita Radix Gentianae grouper fry after infecting
In order to ensure the larva subinfection again stimulating cryptonucleus insect encapsulation to hatch that Margarita Radix Gentianae grouper fry is not come off, within every 2 days, being replaced by seed in clean pond and cultivate, matched group fingerling changes pond by same method.Method particularly includes: the sealing dividing plate in the middle of alternate culture pond is extracted, adopt food calling and drive method whole fries are focused on the opposite side in pond, plug sealing dividing plate, add betagen solution (available iodine content is 10%) the prevention secondary infection of final concentration of 2ppm.Emptying was breeded fish the sea water of side originally, adds the deep fresh water of 30cm, soaks 8 hours, can kill all stimulation cryptonucleus insect encapsulations adhered at the bottom of pond, scrub rear emptying fresh water, fill clean sea water and treat to use next time.Rotation cultivates 5 times as stated above, and the stimulation cryptonucleus insect on fish body all comes off, the temperature control method that whole rotation breeding process all keeps with the above.Hereafter the fry of immunity is all put in clean culturing pool and cultivate, it is not necessary to temperature control, keep natural temperature.
4, the mensuration of the mensuration of fatal dose and effective immune period
The mensuration of Margarita Radix Gentianae cabrilla fatal dose carries out at counteracting toxic substances the last fortnight.Take 50 tail fishes at random from blank group, be divided into 10 groups, often organize 5 tails, separately feed in Plastic Drum.In larva de-bag peak period, collect and 2 hours Nei Tuobao of counting enliven larva, infect 10 groups of fishes with the dose gradient set respectively, infecting water consumption is 1L sea water/tail fish.Each death toll organizing fish in adding up one week, the minimum infective dose that 5 tail Margarita Radix Gentianae cabrillas can be made all dead is exactly the stimulation cryptonucleus insect fatal dose of this fish.
Counteracting toxic substances: every 30 days from matched group and often group immunity cross fry take 50 tail fishes, with the cryptocaryon irritans larva counteracting toxic substances of fatal dose.Together with during counteracting toxic substances, blank matched group is raised together with experimental group, sea water consumption is still every tail fish 1L, carries out infecting counteracting toxic substances with the larva of fatal dose simultaneously, adds up each experimental group real in 7 days
Test the death condition of fish, 9 months interior relative protection ratios of Computation immunity, after stimulating cryptonucleus insect counteracting toxic substances, test the relative protection ratio (RPS) of fish, RPS=(matched group mortality rate-immune group mortality rate)/matched group mortality rate × 100%.
Found that: compared with the matched group not having immunity to cross; the Margarita Radix Gentianae grouper fry mortality rate that immunity is crossed significantly reduces; wherein the 8th group (30 DEG C of infection; 27 DEG C of cultivation) effectively immune period is the longest; the relative protection ratio that after immunity, 8th month also has 58%, the relative protection ratio still having 32% on the 9th month.Therefore, 30 DEG C of infection, the method for 27 DEG C of cultivation can allow Margarita Radix Gentianae cabrilla obtain the longest effective immune period, it is possible to takes the method to produce the Margarita Radix Gentianae grouper fry of irritation cryptonucleus insect.
Advantages of the present invention and effect: 1, operation is simple for infection and rotation cultivation, and do not damage fish body, without fishing for fry during infection, also have only to during rotation cultivation extract the sealing dividing plate in the middle of pond, adopt food calling and drive method and just whole fries can be focused on the opposite side in pond, the method to fish body without any damage, and simple.2. adopt unique alternating temperature to infect and rotation cultural method can allow Margarita Radix Gentianae cabrilla obtain the longest effective immune period; adopt 30 DEG C of infection; the methods of 27 DEG C of cultivation can allow the Margarita Radix Gentianae cabrilla relative protection ratio that 8th month also has 58% after immunity, the relative protection ratio still having 32% on the 9th month.The method of effective immune period any report than before that the method produces will be grown.
Detailed description of the invention
1. the collection and the concentration that stimulate cryptonucleus insect larva alive calculate
Utilize special stimulation cryptonucleus insect legacy system to stimulating cryptonucleus insect go down to posterity and collect, its host of going down to posterity is for avette silvery pomfret, gather well-developed stimulation cryptonucleus insect encapsulation, be placed in sterilized clean beaker and hatch, Spawning water be autoclaved, add the nature seawater of appropriate penicillin and streptomycin；Collecting and hatch 2 hours interior cryptocaryon irritans larva liquid, fully take 50 μ L worm liquid after mixing, add 10 μ L formaldehyde and killed by larva, count under the microscope, note calculates the worm concentration alive of larva liquid.
2. Margarita Radix Gentianae grouper fry is raised and train, is infected
Purchase the Margarita Radix Gentianae grouper fry of about 13g, infect first 2 weeks and fry is carried out domesticating of examination, until its feed, activity are normally, in whole breeding process, raising the use water strainer filtering of 600 orders, its salinity is about 30 ‰, pH is about 8, and dissolved oxygen amount is at more than 5.6mg/L；Margarita Radix Gentianae grouper fry is divided into matched group and experimental group (1-9 group) totally ten groups, experimental group is often organized fry and is infected with hatching 2 hours interior cryptocaryon irritans larvas, during infection water temperature 1-3 group be 24 DEG C, 4-6 group be 27 DEG C, 7-9 group be 30 DEG C, infecting water consumption is 1L sea water/tail fish, and infective dose is 8000 larvas/tail fish；Respectively fry being put in different alternate culture ponds after infecting 6 hours, wherein the 1st, 4,7 groups of water temperatures control at 24 DEG C, and the 2nd, 5,8 groups of water temperatures control at 27 DEG C, and the 3rd, 6,9 groups of water temperatures control at 30 DEG C；Matched group does not carry out stimulating cryptonucleus insect to infect, and temperature of cultivation is 27 DEG C.Alternate culture pond length 40 meters, wide 1.5 meters, high 1 meter, centre, pond seals dividing plate and is divided into two by culturing pool, ensureing both sides water not intercommunication, metainfective fry is totally placed in the side in pond, adds betagen solution (available iodine content is 10%) the prevention secondary infection of final concentration of 2ppm.Concrete packet situation is shown in following table:
| Group number | Infect water temperature (DEG C) | Temperature of cultivation (DEG C) |
| Matched group | It is uninfected by | 27 |
| 1 | 24 | 24 |
| 2 | 24 | 27 |
| 3 | 24 | 30 |
| 4 | 27 | 24 |
| 5 | 27 | 27 |
| 6 | 27 | 30 |
| 7 | 30 | 24 |
| 8 | 30 | 27 |
| 9 | 30 | 30 |
3, the rotation cultivation of Margarita Radix Gentianae grouper fry after infecting
In order to ensure the larva subinfection again stimulating cryptonucleus insect encapsulation to hatch that Margarita Radix Gentianae grouper fry is not come off, within every 2 days, being replaced by seed in clean pond and cultivate, matched group fingerling changes pond by same method.Method particularly includes: the sealing dividing plate in the middle of alternate culture pond is extracted, adopt food calling and drive method whole fries are focused on the opposite side in pond, plug sealing dividing plate, add betagen solution (available iodine content is 10%) the prevention secondary infection of final concentration of 2ppm.Emptying was breeded fish the sea water of side originally, adds the deep fresh water of 30cm, soaks 8 hours, can kill all stimulation cryptonucleus insect encapsulations adhered at the bottom of pond, scrub rear emptying fresh water, fill clean sea water and treat to use next time.Rotation cultivates 5 times as stated above, and the stimulation cryptonucleus insect on fish body all comes off, the temperature control method that whole rotation breeding process all keeps with the above.Hereafter the fry of immunity is all put in clean culturing pool and cultivate, it is not necessary to temperature control, keep natural temperature.
4, the mensuration of the mensuration of fatal dose and effective immune period
The mensuration of Margarita Radix Gentianae cabrilla fatal dose carries out at counteracting toxic substances the last fortnight.Take 50 tail fishes at random from blank group, be divided into 10 groups, often organize 5 tails, separately feed in Plastic Drum.In larva de-bag peak period, collect and 2 hours Nei Tuobao of counting enliven larva, infect 10 groups of fishes with the dose gradient set respectively, infecting water consumption is 1L sea water/tail fish.Each death toll organizing fish in adding up one week, the minimum infective dose that 5 tail Margarita Radix Gentianae cabrillas can be made all dead is exactly the stimulation cryptonucleus insect fatal dose of this fish.
Counteracting toxic substances: every 30 days from matched group and often group immunity cross fry take 50 tail fishes, with the cryptocaryon irritans larva counteracting toxic substances of fatal dose.Together with during counteracting toxic substances, blank matched group is raised together with experimental group; sea water consumption is still every tail fish 1L; carry out infecting counteracting toxic substances with the larva of fatal dose simultaneously; add up each experimental group and in 7 days, test the death condition of fish; 9 months interior relative protection ratios of Computation immunity; the relative protection ratio (RPS) of fish, RPS=(matched group mortality rate-immune group mortality rate)/matched group mortality rate × 100% is tested after stimulating cryptonucleus insect counteracting toxic substances.
Found that: compared with the matched group not having immunity to cross; the Margarita Radix Gentianae grouper fry mortality rate that immunity is crossed significantly reduces; wherein the 8th group (30 DEG C of infection; 27 DEG C of cultivation) effectively immune period is the longest; the relative protection ratio that after immunity, 8th month also has 58%, the relative protection ratio still having 32% on the 9th month.Therefore, 30 DEG C of infection, the method for 27 DEG C of cultivation can allow Margarita Radix Gentianae cabrilla obtain the longest effective immune period, it is possible to takes the method to produce the Margarita Radix Gentianae grouper fry of irritation cryptonucleus insect.
Claims (1)
1. a production method for irritation cryptonucleus insect Margarita Radix Gentianae grouper fry, including following four step:
(1) collection and the concentration that stimulate cryptonucleus insect larva alive calculate
Utilize special stimulation cryptonucleus insect legacy system to stimulating cryptonucleus insect go down to posterity and collect, its host of going down to posterity is for avette silvery pomfret, gather well-developed stimulation cryptonucleus insect encapsulation, be placed in sterilized clean beaker and hatch, Spawning water be autoclaved, add the nature seawater of appropriate penicillin and streptomycin；Collecting and hatch 2 hours interior cryptocaryon irritans larva liquid, fully take 50 μ L worm liquid after mixing, add 10 μ L formaldehyde and killed by larva, count under the microscope, note calculates the worm concentration alive of larva liquid.
(2) Margarita Radix Gentianae grouper fry is raised and train, is infected
Purchase the Margarita Radix Gentianae grouper fry of about 13g, infect first 2 weeks and fry is carried out domesticating of examination, until its feed, activity are normally, in whole breeding process, raising the use water strainer filtering of 600 orders, its salinity is about 30 ‰, pH is about 8, and dissolved oxygen amount is at more than 5.6mg/L；Margarita Radix Gentianae grouper fry is divided into matched group and experimental group (1-9 group) totally ten groups, experimental group is often organized fry and is infected with hatching 2 hours interior cryptocaryon irritans larvas, during infection water temperature 1-3 group be 24 DEG C, 4-6 group be 27 DEG C, 7-9 group be 30 DEG C, infecting water consumption is 1L sea water/tail fish, and infective dose is 8000 larvas/tail fish；Respectively fry being put in different alternate culture ponds after infecting 6 hours, wherein the 1st, 4,7 groups of water temperatures control at 24 DEG C, and the 2nd, 5,8 groups of water temperatures control at 27 DEG C, and the 3rd, 6,9 groups of water temperatures control at 30 DEG C；Matched group does not carry out stimulating cryptonucleus insect to infect, and temperature of cultivation is 27 DEG C.Alternate culture pond length 40 meters, wide 1.5 meters, high 1 meter, centre, pond seals dividing plate and is divided into two by culturing pool, ensureing both sides water not intercommunication, metainfective fry is totally placed in the side in pond, adds betagen solution (available iodine content is 10%) the prevention secondary infection of final concentration of 2ppm.
(3) the rotation cultivation of Margarita Radix Gentianae grouper fry after infecting
In order to ensure the larva subinfection again stimulating cryptonucleus insect encapsulation to hatch that Margarita Radix Gentianae grouper fry is not come off, within every 2 days, being replaced by seed in clean pond and cultivate, matched group fingerling changes pond by same method；Method particularly includes: the sealing dividing plate in the middle of alternate culture pond is extracted, adopt food calling and drive method whole fries are focused on the opposite side in pond, plug sealing dividing plate, add betagen solution (available iodine content is 10%) the prevention secondary infection of final concentration of 2ppm；Emptying was breeded fish the sea water of side originally, adds the deep fresh water of 30cm, soaks 8 hours, can kill all stimulation cryptonucleus insect encapsulations adhered at the bottom of pond, scrub rear emptying fresh water, fill clean sea water and treat to use next time；Rotation cultivates 5 times as stated above, and the stimulation cryptonucleus insect on fish body all comes off, the temperature control method that whole rotation breeding process all keeps with the above；Hereafter the fry of immunity is all put in clean culturing pool and cultivate, it is not necessary to temperature control, keep natural temperature.
(4) the measuring and the mensuration of effective immune period of fatal dose
The mensuration of Margarita Radix Gentianae cabrilla fatal dose carries out at counteracting toxic substances the last fortnight, takes 50 tail fishes at random from blank group, is divided into 10 groups, often organizes 5 tails, separately feeds in Plastic Drum.In larva de-bag peak period, collect and 2 hours Nei Tuobao of counting enliven larva, infect 10 groups of fishes with the dose gradient set respectively, infecting water consumption is 1L sea water/tail fish.Each death toll organizing fish in adding up one week, the minimum infective dose that 5 tail Margarita Radix Gentianae cabrillas can be made all dead is exactly the stimulation cryptonucleus insect fatal dose of this fish；
Counteracting toxic substances: every 30 days from matched group and often group immunity cross fry take 50 tail fishes, with the cryptocaryon irritans larva counteracting toxic substances of fatal dose.Together with during counteracting toxic substances, blank matched group is raised together with experimental group; sea water consumption is still every tail fish 1L; carry out infecting counteracting toxic substances with the larva of fatal dose simultaneously; add up each experimental group and in 7 days, test the death condition of fish; 9 months interior relative protection ratios of Computation immunity; the relative protection ratio (RPS) of fish, RPS=(matched group mortality rate-immune group mortality rate)/matched group mortality rate × 100% is tested after stimulating cryptonucleus insect counteracting toxic substances；
Finally, the measurement result according to effective immune period, select to allow Margarita Radix Gentianae cabrilla obtain the alternating temperature infection of the longest effective immune period, cultural method, produce the Margarita Radix Gentianae grouper fry of irritation cryptonucleus insect；Adopt the method can produce the Margarita Radix Gentianae grouper fry of the immune protective efficiency irritation cryptonucleus insect up to more than 8 months.
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| CN201610236360.0A CN105746410A (en) | 2016-04-08 | 2016-04-08 | Production method of cryptocaryon irritus-resistant epinephelus fuscoguttatus and epinephelus lanceolatus fry |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112889717A (en) * | 2021-02-24 | 2021-06-04 | 中山大学 | Method for biologically preventing and controlling cryptocaryon irritans infection by using tilapia |
| CN115474564A (en) * | 2022-09-07 | 2022-12-16 | 华南农业大学 | Method for breeding epinephelus coioides |
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2016
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112889717A (en) * | 2021-02-24 | 2021-06-04 | 中山大学 | Method for biologically preventing and controlling cryptocaryon irritans infection by using tilapia |
| CN112889717B (en) * | 2021-02-24 | 2022-03-15 | 中山大学 | Method for biologically preventing and controlling cryptocaryon irritans infection by using tilapia |
| CN115474564A (en) * | 2022-09-07 | 2022-12-16 | 华南农业大学 | Method for breeding epinephelus coioides |
| CN115474564B (en) * | 2022-09-07 | 2023-10-20 | 华南农业大学 | Breeding method of Epinephelus coioides |
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Application publication date: 20160713 |