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Fish-derived vibrio parahaemolyticus controlled-release oral vaccine and preparation and utilization method

A technology of Vibrio hemolyticus and oral vaccine, which is applied to the vaccine and application field of farmed fish, can solve the problems of complex microsphere vaccine process, difficult to popularize and apply on a large scale, low content of outer membrane protein, etc., and achieves high market application value, Overcome ineffective, easy-to-use effects

Pending Publication Date: 2019-03-12
天津市水产研究所
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] The deficiencies that above-mentioned two methods exist are: method (1) adopt medicinal starch as carrier, take acrylic resin as coating material to prepare Vibrio parahaemolyticus, Vibrio alginolyticus and Edwardsiella tarda triple oral vaccine, food-attracting effect relatively poor; method (2) needs to extract the outer membrane protein, and the outer membrane protein content is low, and it is difficult to extract, and the microsphere vaccine of the outer membrane protein prepared by the emulsification method is complicated, time-consuming and labor-intensive, and the production cost is high, so it is difficult to Scale promotion and application

Method used

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  • Fish-derived vibrio parahaemolyticus controlled-release oral vaccine and preparation and utilization method
  • Fish-derived vibrio parahaemolyticus controlled-release oral vaccine and preparation and utilization method
  • Fish-derived vibrio parahaemolyticus controlled-release oral vaccine and preparation and utilization method

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Experimental program
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Effect test

preparation example Construction

[0036] The preparation of fish source Vibrio parahaemolyticus slow-release oral vaccine, the method of this preparation process is as follows:

[0037] (1) Inactivation: Use 0.5% formaldehyde to inactivate the Vibrio parahaemolyticus liquid;

[0038] ⑵ Vaccine preparation:

[0039] According to commercial feed: the concentration is 6×10 10 The Vibrio parahaemolyticus ratio of cfu / mL is: 10:1, and the unit is: gram:mL. Put the commercialized feed into the coating machine, start the turntable, and the pellet machine adopts a rotating speed of 400 rpm to inactivate The Vibrio parahaemolyticus vaccine is slowly dripped into the center of the turntable, and the vaccine is sprayed onto the feed by the centrifugal force of the central turntable to prepare vaccine feed. The effective concentration of the vaccine is about 6×10 9 cfu / g feed, put it into an oven for drying at 37°C for later use.

[0040] ⑶ Sustained release layer coating preparation:

[0041] 1. Sustained-release age...

example 1

[0049] The vaccine feed prepared above is used to immunize turbot, and the specification of turbot is 102±11.2 grams. The amount of feeding was the same as that of normal bait. Normal bait was fed for 7 consecutive days, with an interval of 14 days to boost immunity for 7 days.

[0050] In the experiment of immunizing turbot in small water body of aquarium, two groups were designed, the control group and the slow-release oral immunization group, with 3 repetitions in each group. The slow-release oral immunization group was fed the prepared slow-release oral vaccine twice a day for 7 consecutive days, and then fed normal bait; the control group was fed normal granule bait. Blood was collected once every 7 days, and three fish were collected in each group, and the antibody level in the serum was measured by enzyme-linked immunosorbent assay (Elisa). After 28 days, test fish and control fish were subjected to Vibrio parahaemolyticus (concentration of bacteria liquid was 3×10 8 ...

example 2

[0055] Workshop breeding test:

[0056] Breeding mode: factory farming; farmed fish: flounder, size 89±15.2 grams; a total of 6 test ponds, including 3 test ponds and 3 control ponds. First immunization: feed twice a day for 7 consecutive days; booster immunization: 21 days after the first immunization, feed twice a day for 7 days. The control pond was fed with common bait, and the other breeding and management methods were the same. After 2 months, the serum of 5 fish was collected from each pond, and the antibody titer was determined by agglutination method, and 50 flounder were collected for Vibrio parahaemolyticus (the concentration of bacteria solution was 3×10 8 cfu / mL) in the artificial infection experiment, each fish was injected with 0.2 mL, the mortality within 14 days was counted, and the immune protection rate was calculated. The test results are shown in Table 3.

[0057] Table 3 Serum antibody titer and immune protection rate of flounder

[0058]

[0059] ...

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Abstract

The invention relates to a preparation and utilization method of a fish-derived vibrio parahaemolyticus controlled-release oral vaccine. The controlled-release oral vaccine is prepared from feed, inactivated vibrio parahaemolyticus liquid and a controlled release agent according to a ratio of 10:1:3. The preparation method includes steps: (1) vibrio parahaemolyticus culture and inactivation; (2) oral vaccine preparation; (3) controlled-release oral vaccine preparation. The utilization method includes steps of (1) primary immunization and (2) booster immunization. By the fish-derived vibrio parahaemolyticus controlled-release oral vaccine and an oral immunization inoculation method, great immunization effects, low workload and suitableness for large-scale inoculation are realized. In addition, by coating of the oral vaccine with the controlled release agent, vaccine release time can be effectively prolonged, resistance of antigens to damages and decomposition by digestive enzymes in thestomach and foregut is realized, and the antigens are enabled to smoothly reach the hindgut to have immunoreaction, so that great immunization effects are obtained.

Description

technical field [0001] The invention belongs to the technical field of vaccines for cultured fish and their use, and in particular relates to a slow-release oral vaccine of fish-derived Vibrio parahaemolyticus and its preparation and use methods. Background technique [0002] The diseases of marine cultured fish in my country are increasing year by year, especially vibriosis is the most common, causing huge economic losses. At present, the domestic treatment of fish bacterial diseases is mainly through the use of antibiotics, while the prevention of bacterial diseases mainly uses methods such as disinfectants, water quality improvers, and microecological preparations to improve the ecological environment of aquaculture water bodies. The application of vaccines is rare. The main reason is that there are very few aquatic vaccines that have been officially approved, and the vaccines that have been officially approved in China are all injection-type vaccines. The fish body is u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61K9/52A61K47/38A61K47/12A61P31/04
CPCA61K9/0053A61K9/5015A61K9/5047A61K39/107A61K2039/521A61K2039/542A61K2039/55A61P31/04
Inventor 张振国罗璋郝爽冯守明耿绪云史谢尧尤宏争
Owner 天津市水产研究所
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