CN1331534C - Aquaculture creature disease controlling compound vaccine and its preparation method - Google Patents

Aquaculture creature disease controlling compound vaccine and its preparation method Download PDF

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CN1331534C
CN1331534C CNB2005100958149A CN200510095814A CN1331534C CN 1331534 C CN1331534 C CN 1331534C CN B2005100958149 A CNB2005100958149 A CN B2005100958149A CN 200510095814 A CN200510095814 A CN 200510095814A CN 1331534 C CN1331534 C CN 1331534C
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vibrio
vaccine
aeromonas
plesiomonas
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CN1762387A (en
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刘为国
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Dalian University
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Dalian University
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Abstract

The present invention relates to composite bacterial vaccine for preventing aquiculture animal diseases, which is mainly composed of vibrio alginolyticus, vibrio parahaemolyticus, vibrio anguillarum, aeromonas hydrophila, temperate aeromonas, pseudomonas aeruginosa and Shigella plesiomonas. The contents of each strain in the composite bacterial vaccine comprise: 10 to 50% of vibrio alginolyticus, 10 to 50% of aeromonas hydrophila, and 0 to 50 of other strains. The total amount of bacteria is 1*10<7> to 1*10<10> number/ml. The composite bacterial vaccine has the manufacturing method that each single bacterium is respectively cultivated, inactivated, washed and diluted, and the composite bacterial vaccine is prepared proportionally. Compared with the prior art, the composite bacterial vaccine of the present invention can widely prevent common bacteriosis during applying in the process of aquiculture animals, and especially has satisfactory effect on preventing bacteriosis, such as shrimps, fishes, sea cucumbers, etc., of mariculture.

Description

The compound vaccine and the method for making thereof of control aquaculture creature disease
Technical field
The present invention relates to a kind of compound vaccine of preventing and treating the aquaculture disease.
Background technology
Bacterial disease is the common disease in the sea-farmings such as prawn, Fish, Stichopus japonicus, as: red appendages disease of prawn (erythromelia) cause of disease has: vibrio parahaemolytious, Vibrio anguillarum, vibrio alginolyticus: mashed oculopathy cause of disease: non-01 group cholera vibrio: the gill rot cause of disease has: vibrio, Rhodopseudomonas, Aeromonas; The brown spot cause of disease has: vibrio, pseudomonas, spirillum, Flavobacterium; The Aeromonas disease pathogen has: Aeromonas: the vibriosis cause of disease of prawn germling has: Vibrio anguillarum, vibrio parahaemolytious, vibrio alginolyticus; And these antibacterials also can cause the generation of culture diseases such as Fish, Stichopus japonicus, as: the Peptic Ulcers of marine fish, gill rot etc.; The skin of Stichopus japonicus rots limit, ulcer etc.These bacterial diseasees often cause the large tracts of land death in the sea-farming, culture to Aquatic product and bring great loss.In the raising fish and shrimp process, in a single day manifest symptom appears in the fish and shrimp disease, has been the course of disease later stage promptly, is difficult to treatment.Therefore mainly be putting prevention first for the fish and shrimp disease, and the disinfectant and the antibiotic that use at present exist drug residue, develop immunity to drugs, pollute a lot of drawbacks such as breeding environment.
Therefore, immunological method becomes control aquatic animal fulminant epidemic diseases optimum selection.
In existing technology, also has weak point for the control of fish and shrimp disease, as " vibrio vaccine is to the effect of Penaeus monodon and japonicus immunoprophylaxis " aquatic product journal .2000.024 (006) .-564-569.Attempt using immunologic method to prevent and treat vibriosis penaeus.The cause of disease vibrio parahaemolytious is made formalin-inactivated vaccine; under experiment condition respectively with soak, mode such as oral and injection is inoculated in Penaeus monodon and japonicus; found that in certain scope, the immune protective rate of prawn raises with the increase of vaccine concentration.Can only prevent and treat vibriosis penaeus but exist, the single shortcoming of disease preventing and treating kind.Calendar year 2001 JIUYUE to disclose application number on the 5th be 00114083.3 patent " mixed vaccine of a kind of anti-aquatic animal vibriosis and preparation method thereof ".This mixed vaccine is obtained for two kinds by Vibrio vulnificus, vibrio alginolyticus, vibrio parahaemolytious, vibrio tenuis, Vibrio flurialis, Kazakhstan vibrio and Vibrio anguillarum etc. and mixes more than two kinds, can prevent and treat the invasion and attack of multiple vibrio.But the defense reaction to other pathogenic infections is relatively poor, and bacterial disease relative complex in the sea-farming practice, as vibriosis, Aeromonas disease etc., so its practical application is restricted.Disclosed application number is that 02110261.9 " a kind of process for preparing bacterial vaccine of aquatic animal " is though related to multiple pathogenetic bacteria, but it is mainly used in mammals such as fish or dolphin, does not explain for the protection effect of zootic diseases such as prawn." the seawater fish pathogenic vibrio is to the pathogenicity of prawn and the immunoprophylaxis of vaccine thereof " tropical oceanography is reported .2001.020 (004) .-68-73.Fish morbid vibrio (vibrio alginolyticus, Vibrio vulnificus, the biological I type of Vibrio flurialis, Vibrio anguillarum and vibrio parahaemolytious) can make Penaeus monodon Penaeus monodon ill equally, only to the pathogenic of Penaeus monodon and immunoprotection masterpiece research.By the intramuscular injection infection experiment, find that 5 kinds of vibrios all have different pathogenic effects to Penaeus monodon.From 5 kinds of vibrios, filter out virulent strain, make formalin-inactivated vaccine with it.By injection, dipping bath and 3 kinds of Penaeus monodons that mode is inoculated such as throw something and feed, except that the DeGrain of throwing something and feeding, injection and dipping bath are inoculated all certain immune protective efficiency.In the injection inoculation group, along with the raising of vaccine concentration, its immunity has trend of rising.Explanation is in the vibriosis of fish and shrimp, and vaccine can play common protection effect, for the production of commercial fish and shrimp vaccine provides may.Though injection and dipping bath inoculation have certain immune protective efficiency, for aquaculture, ill fish and shrimp are caught out inject and dipping bath is inoculated, almost be impossible thing.
Summary of the invention
The object of the present invention is to provide the multiple aquatic animal of a kind of immunity, the compound vaccine that the defence various pathogens infects is to overcome the existing problem of present vaccine.
The present invention makes compound vaccine control aquaculture disease with the pathogenic bacterium in the common sea-farming, it is characterized in that mainly being mixed by one or more vibrios in the following antibacterial and one or more other antibacterials except that vibrio, described vibrio strain is selected from: vibrio alginolyticus (V.alginolyticus), vibrio parahaemolytious (V.parahaemolyticus), Vibrio anguillarum (V.anguillarum); Described other bacteria cultures except that vibrio are selected from: Aeromonas hydrophila (A.hydrophila), Aeromonas sobria (A.sobria), Pseudomonas aeruginosa (P.aeruginosa), class will Hayes class will He Plesiomonas (P.shigelloides).Each composition vaccine content is in the compound vaccine: vibrio alginolyticus, Aeromonas hydrophila are 10~50%; Vibrio parahaemolytious, Vibrio anguillarum, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas are 0~50%, and bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml.
According to above principle, can be respectively through the proportioning in the preferred compound vaccine:
1, vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas content are 10~20%.
2, vibrio alginolyticus and Aeromonas hydrophila respectively are 50%.
3, vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum and Aeromonas hydrophila are 10~50%.
4, vibrio alginolyticus and Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas account for 10~50% respectively.
Specifically, below the compound vaccine prevention effect of prescription is quite excellent:
1, the polyvalent vaccine of vibrio alginolyticus 10%, vibrio parahaemolytious 10%, Vibrio anguillarum 10%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 20%, class will He Plesiomonas 20% composition, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml.
2, the polyvalent vaccine of vibrio alginolyticus 15%, vibrio parahaemolytious 15%, Vibrio anguillarum 15%, Aeromonas hydrophila 10%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 20% composition, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml
3, the polyvalent vaccine of vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% composition, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml.
4, the polyvalent vaccine of vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 15%, Aeromonas sobria 15%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% composition, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml.
The manufacture method of compound vaccine
1, the single component vaccine for preparing vibrio and other antibacterials respectively:
(1) vibrio and other bacteria cultures are inoculated in culture medium, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, physics or chemosterilization.
(3) 3000~5000 rev/mins centrifugal, removes impurity.
(4) be mixed with desired concn with normal saline, promptly get the single component vaccine.
2, the single component vaccine that makes is mixed mutually by the concentration of formula proportion promptly get described compound vaccine.
3, above-mentioned compound vaccine being added suitable preservatives preserves standby.
Compared with prior art, compound vaccine of the present invention can be prevented and treated common bacterial disease more widely in being applied to the aquatic animal process, particularly to the bacterial disease control of the shrimp of sea-farming, fish, Stichopus japonicus etc., obtains satisfied effect.
Below in conjunction with embodiment the present invention is described.
The specific embodiment
Embodiment one:
Vibrio alginolyticus and Aeromonas hydrophila respectively account for producing of 50% polyvalent vaccine of forming.
1. the single component vaccine for preparing vibrio alginolyticus and Aeromonas hydrophila respectively:
(1) vibrio alginolyticus and Aeromonas hydrophila strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 1~5% formalin deactivation.
(3) physiological saline solution is used in 3000~5000 rev/mins of centrifugal backs of culture of deactivation, centrifugal again, repeated multiple times is to remove remaining formalin.
(4) with the precipitate of the last centrifugal gained of physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that makes is respectively accounted for 50% proportional concentration by prescription and mix mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
Embodiment two:
Vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum and Aeromonas hydrophila account for producing of 25% polyvalent vaccine of forming respectively.
1. the single component vaccine for preparing vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum and Aeromonas hydrophila respectively:
(1) vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum and Aeromonas hydrophila strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 1~5% formalin deactivation.
(3) use physiological saline solution behind culture 3000~5000 commentaries on classics part Zhongli's heart with deactivation, centrifugal again, repeated multiple times is to remove remaining formalin.
(4) with the precipitate of the last centrifugal gained of physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that makes is accounted for 25% proportional concentration respectively by prescription and mix mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
Embodiment three:
Vibrio alginolyticus and Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas account for 20% polyvalent vaccine of forming respectively.
1. the single component vaccine for preparing vibrio alginolyticus and Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas respectively:
(1) vibrio alginolyticus and Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 1~5% formalin deactivation.
(3) physiological saline solution is used in 3000~5000 rev/mins of centrifugal backs of culture of deactivation, centrifugal again, repeated multiple times is to remove remaining formalin.
(4) with the precipitate of the last centrifugal gained of physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that makes is accounted for 20% proportional concentration respectively by prescription and mix mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
Embodiment four:
The polyvalent vaccine that vibrio alginolyticus 10%, vibrio parahaemolytious 10%, Vibrio anguillarum 10%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 20%, class will He Plesiomonas 20% are formed.
1. the single component vaccine for preparing vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas respectively:
(1) vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 1~5% formalin deactivation.
(3) physiological saline solution is used in 3000~5000 rev/mins of centrifugal backs of culture of deactivation, centrifugal again, repeated multiple times is to remove remaining formalin.
(4) with the precipitate of the last centrifugal gained of physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that the makes proportional concentration by prescription vibrio alginolyticus 10%, vibrio parahaemolytious 10%, Vibrio anguillarum 10%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 20%, class will He Plesiomonas 20% is mixed mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
Embodiment five:
The polyvalent vaccine that vibrio alginolyticus 15%, vibrio parahaemolytious 15%, Vibrio anguillarum 15%, Aeromonas hydrophila 10%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 20% are formed.
1. the single component vaccine for preparing vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas respectively:
(1) vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 1~5% formalin deactivation.
(3) physiological saline solution is used in 3000~5000 rev/mins of centrifugal backs of culture of deactivation, centrifugal again, repeated multiple times is to remove remaining formalin.
(4) with the precipitate of the last centrifugal gained of physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that the makes proportional concentration by prescription vibrio alginolyticus 15%, vibrio parahaemolytious 15%, Vibrio anguillarum 15%, Aeromonas hydrophila 10%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 20% is mixed mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
Embodiment six:
The polyvalent vaccine that vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% are formed.
1. the single component vaccine for preparing vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas respectively:
(1) vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 1~5% formalin deactivation.
(3) physiological saline solution is used in 3000~5000 rev/mins of centrifugal backs of culture of deactivation, centrifugal again, repeated multiple times is to remove remaining formalin.
(4) with the precipitate of the last centrifugal gained of physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that the makes proportional concentration by prescription vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% is mixed mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
Embodiment seven:
The polyvalent vaccine that vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 15%, Aeromonas sobria 15%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% are formed.
1. the single component vaccine for preparing vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas respectively:
(1) vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 1~5% formalin deactivation.
(3) physiological saline solution is used in 3000~5000 rev/mins of centrifugal backs of culture of deactivation, centrifugal again, repeated multiple times is to remove remaining formalin.
(4) with the precipitate of the last centrifugal gained of physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that the makes proportional concentration by prescription vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 15%, Aeromonas sobria 15%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% is mixed mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
Embodiment eight:
The polyvalent vaccine that vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 15%, Aeromonas sobria 15%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% are formed.
1. the single component vaccine for preparing vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas respectively:
(1) vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas strain are inoculated in culture medium respectively, cultivated 24~48 hours for 25~37 ℃.
(2) normal saline washing culture, 56 ℃~65 ℃ water-bath deactivations 60~120 minutes.
(3) physiological saline solution is used in 3000~5000 rev/mins of centrifugal backs of culture of deactivation, centrifugal again, repeated multiple times is to remove impurity.
(4) be mixed with desired concn with physiological saline solution, promptly get the single component vaccine.
2. the single component vaccine that the makes proportional concentration by prescription vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 15%, Aeromonas sobria 15%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15% is mixed mutually, bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml, promptly get described polyvalent vaccine.
3. above-mentioned compound vaccine being added 1% benzyl alcohol preserves standby as antiseptic.
This compound vaccine is mainly used in the common bacterial disease in control the aquaculture disease, particularly sea-farming such as prawn, Fish.These bacterial diseasees often cause the large tracts of land death in the sea-farming, use and have obtained satisfied effect behind this compound vaccine, are exemplified below:
1, in JIUYUE in 1997 27 days, cultures the sea area from the Pulandian and fetch 16 of the long prawns of average 5cm, feed in the laboratory glass jar.The prawn of fetching all suffers from brown spot in various degree, embodiment five vaccines 2~3 times of throwing something and feeding through every day, and all the other 14 through desquamation repeatedly in natural death in November then except that two, and survival is in August, 1998, about the long 16cm of shrimp body.
2, on July 5th, 1999, find 4 female prawn appendages color that reddens, move about slowly, be reluctant feed.Satisfy to adjust using two vaccine every days of embodiment 2 times is 3~4 times, and through treatment in 4 days, morbid state disappeared, and survival is to natural death in January, 2000, and body reaches 18cm.
3, in June, 1999, the field application test of this product is carried out in one 100 mu shrimp pond in shrimp field, Pulandian City.After finding that by the end of June the part shrimp is ill, bring into use embodiment seven vaccines by 2 medications weekly, use always to 9 the end of month prawn go out the pond, again the morbidity.The shrimp that closes on the contrast pond then falls ill constantly, and output is 110 jin/hundred mu.Serious because of shrimp disease in 1999, test tank output is 70~80 jin/mu then, and shrimp body stalwartness.No matter from the individual height of prawn, weight, still per mu yield, after using vaccine, curative effect all is better than matched group.
4, this test in May, 2003 to 2003 year JIUYUE carry out in coastal shrimp field, stone river town head shop, Pulandian City, Liaoning Province village, this place has 12 shrimp ponds, choose and be ready 4 shrimp ponds of Application Example eight compound vaccines as test group, all the other 8 shrimp ponds are matched group.Disinfect with tea seed cake in the shrimp pond before throwing Seedling.Throw in japonicus shrimp Seedling, going into pond shrimp Seedling specification is 0.9~1.0cm, and 50~100 mu of cultured areas are thrown in shrimp Seedling 40~800,000 tails.Test group prawn shell is clean and tidy, bright, on average survives 102 days, manys growth 40.75 days than matched group, and prawn is ripe more, and therefore, the average height comparison of test group prawn is according to group leader 4.0cm.
5, successively repeatedly used nearly 100 to view and admire in 2003~2004 years, do not change water in nearly 50 days, breeding environment is worsened with Carassius auratus researching fish anti-infection ability, reaching has Carassius auratus death, changes water then, uses embodiment six compound vaccines simultaneously, through rescuing, there are 86 Carassius auratuses to survive.
6, fish culture experiment: buy and suffer from 15 of Peptic Ulcers Carassius auratuss,, 12 Peptic Ulcers recoveries from illness are arranged through using embodiment three compound vaccines.
7, the laboratory of Stichopus japonicus is cultured: each 100 young ginseng of experimental group and matched group, and use embodiment two vaccines to culture through 1 year, experimental group survives 76, and matched group only survives 36.Ulcer to Stichopus japonicus has also demonstrated certain prevention effect.

Claims (10)

1, a kind of compound vaccine of preventing and treating aquaculture creature disease, it is characterized in that mainly being mixed by one or more vibrios and one or more other antibacterials except that vibrio, described vibrio strain is selected from: vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum; Described other bacteria cultures except that vibrio are selected from: Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas; Each composition vaccine content is in the compound vaccine: vibrio alginolyticus, Aeromonas hydrophila are 10~50%; Vibrio parahaemolytious, Vibrio anguillarum, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas are 0~50%, and bacteria total amount is 1 * 10 7~1 * 10 10Individual/ml.
2,, it is characterized in that the content of vibrio alginolyticus in the described compound vaccine, vibrio parahaemolytious, Vibrio anguillarum, Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas is 10~20% according to the compound vaccine of the described control aquaculture creature disease of claim 1.
3,, it is characterized in that vibrio alginolyticus and Aeromonas hydrophila respectively account for 50% in the described compound vaccine according to the compound vaccine of the described control aquaculture creature disease of claim 1.
4,, it is characterized in that vibrio alginolyticus, vibrio parahaemolytious, Vibrio anguillarum and Aeromonas hydrophila account for 10~50% respectively in the described compound vaccine according to the compound vaccine of the described control aquaculture creature disease of claim 1.
5,, it is characterized in that vibrio alginolyticus and Aeromonas hydrophila, Aeromonas sobria, Pseudomonas aeruginosa, class will He Plesiomonas account for 10~50% respectively in the described compound vaccine according to the compound vaccine of the described control aquaculture creature disease of claim 1.
6, according to the compound vaccine of the described control aquaculture creature disease of claim 2, it is characterized in that each vaccine content is respectively in the described compound vaccine: vibrio alginolyticus 10%, vibrio parahaemolytious 10%, Vibrio anguillarum 10%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 20%, class will He Plesiomonas 20%.
7, according to the compound vaccine of the described control aquaculture creature disease of claim 2, it is characterized in that each vaccine content is respectively in the described compound vaccine: vibrio alginolyticus 15%, vibrio parahaemolytious 15%, Vibrio anguillarum 15%, Aeromonas hydrophila 10%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 20%.
8, according to the compound vaccine of the described control aquaculture creature disease of claim 2, it is characterized in that each vaccine content is respectively in the described compound vaccine: vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 20%, Aeromonas sobria 10%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15%.
9, according to the compound vaccine of the described control aquaculture creature disease of claim 2, it is characterized in that each vaccine content is respectively in the described compound vaccine: vibrio alginolyticus 15%, vibrio parahaemolytious 10%, Vibrio anguillarum 15%, Aeromonas hydrophila 15%, Aeromonas sobria 15%, Pseudomonas aeruginosa 15%, class will He Plesiomonas 15%.
10, a kind of method for making of preventing and treating the compound vaccine of aquaculture creature disease is characterized in that used strain and proportioning are:
Vibrio:
Vibrio alginolyticus 10~50%
Vibrio parahaemolytious 0~50%
Vibrio anguillarum 0~50%
Other antibacterials except that vibrio
Aeromonas hydrophila 10~50%
Aeromonas sobria 0~50%
Pseudomonas aeruginosa 0~50%
Class will He Plesiomonas 0~50%
Processing step is: (1) each strain carries out single culture through inoculation, cultivation in 25~37 ℃, 24~48 hours, normal saline washing, deactivation, centrifugal, the formulated single vaccine of normal saline; (2) by formula proportion each single vaccine is mixed as required and carry out the composite compound vaccine that forms; (3) adding antiseptic preserves.
CNB2005100958149A 2005-08-15 2005-08-15 Aquaculture creature disease controlling compound vaccine and its preparation method Expired - Fee Related CN1331534C (en)

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