CN110694058A - A kind of rabbit coccidiosis live vaccine and its preparation method and application - Google Patents

Abstract

The invention relates to the technical field of animal coccidiosis vaccines, in particular to a live rabbit coccidiosis vaccine and a preparation method and application thereof. The present invention provides a rabbit coccidiosis live vaccine composition, which comprises early-maturing attenuated strains of Eimeria maxima, Eimeria enterica and Eimeria medium, and precocious strains of Eimeria piercing. Weakened strains. The rabbit coccidiosis live vaccine provided by the invention has high immune protection efficacy and safety at the same time, can effectively prevent and treat rabbit coccidiosis, reduce or eliminate the use of anti-coccidiosis drugs, and at the same time significantly reduce the effect of vaccination on the weight gain of rabbits It has good economic benefits and broad application prospects. The invention also provides a method for breeding coccidian early-maturing attenuated strains, which can effectively shorten the breeding cycle of the early-maturing strains, improve the breeding efficiency of the coccidia early-maturing attenuated strains, and the obtained early-maturing strains have stable genetic characteristics, The pathogenicity is significantly reduced and the immunogenicity is good.

Description

A kind of rabbit coccidiosis live vaccine and its preparation method and application

technical field

The invention relates to the technical field of animal coccidiosis vaccines, in particular to a live rabbit coccidiosis vaccine and a preparation method and application thereof.

Background technique

Antibiotics and anticoccidial drugs have a long history as livestock and poultry feed additives, and they are the main means to ensure the health of livestock and poultry under the intensive breeding mode. With the frequent occurrence of drug residues and drug-resistant bacteria (insects) strains and threats to the environment and human health, the strategy of using drugs to prevent and control animal diseases has gradually exposed the drawbacks. Outbreaks of coccidiosis in rabbits can cause large-scale death of rabbits; chronic and subclinical coccidiosis infections lead to reduced animal performance and economic benefits. The continuous occurrence of coccidiosis is the main reason for the low economic efficiency or even loss of many rabbit farms. Therefore, under the predicament of the limited use of anticoccidial drugs and the frequent occurrence of coccidial drug resistance, which leads to poor drug control effects, it is urgent to develop an efficient and safe rabbit coccidiosis vaccine.

The pathogen of rabbit coccidiosis is 11 species of Eimeria, which are parasitic in different parts of the intestine or liver. The pathogenicity and prevalence of different species are different, and the harm to the rabbit industry is different. Among them, Eimeria large, Eimeria medium, Eimeria intestinalis and Eimeria perforating are the main insect species that are infected and prevalent in rabbit farms, and the first three species have strong pathogenic Disease, causing severe diarrhea, seriously affecting the production performance and economic benefits of the rabbit industry. Therefore, the development of rabbit coccidiosis vaccine must achieve two main goals: one is to have a good protective effect on pathogenic coccidiosis species; the other is to have high safety. Vaccines composed of live oocysts can effectively resist re-infection with the same species of coccidioides, showing great advantages in the prevention of coccidiosis in chickens and turkeys. Therefore, the development of live vaccines against popular coccidiosis species for the prevention of coccidiosis in rabbits can significantly reduce the use of anti-coccidial drugs, and ultimately eliminate the use of anti-coccidial drugs, thereby effectively improving food safety, environment and economy. Benefit, the application prospect is very broad.

SUMMARY OF THE INVENTION

In order to solve the problems existing in the prior art, the purpose of the present invention is to provide a live attenuated coccidiosis vaccine with excellent safety and immune protection efficacy and a method for efficient and rapid selection and breeding of precoccidia attenuated strains.

For achieving the above object, technical scheme of the present invention is as follows:

Although the live oocysts of rabbit coccidiosis as the immunogen of the vaccine are more conducive to improving the immune protection efficacy of the vaccine, the risk of adverse effects on animal growth and production is higher. Therefore, how to ensure the immunity of the rabbit coccidia live vaccine Protecting efficacy while improving its safety is a technical problem faced in the development of rabbit coccidiosis vaccines. In the research of rabbit coccidiosis vaccines, highly pathogenic species are often used for inoculation to obtain the immunity of the corresponding species, while low pathogenic or basically non-pathogenic rabbit coccidioids are often neglected. In the process of developing a rabbit coccidiosis vaccine with high immune efficacy and high safety at the same time, it was found that relatively low doses of Eimeria large, Eimeria intestinalis, Eimeria medium, Eimeria yellow, etc. The precocious attenuated strain of highly pathogenic rabbit coccidia is used in combination with relatively high doses of low pathogenic rabbit coccidia such as Eimeria perfringens, Eimeria micro, and Eimeria viridis. Significantly reduce the impact of the vaccine on the growth of rabbits, effectively improve the safety of the vaccine, and at the same time ensure the immune protection efficacy of the vaccine. Further, in order to improve the preparation efficiency of the vaccine, the present invention develops a rapid breeding method for the early-maturing attenuating strains on the basis of the traditional early-maturing pressure selection method, which effectively shortens the passage and screening period of the early-maturing attenuating strains, and obtains The early-maturing attenuated strains have stable genetic characteristics, high safety and good immunogenicity.

Specifically, the technical scheme of the present invention is as follows:

The present invention provides a rabbit coccidiosis live vaccine composition, which comprises at least three early-maturing attenuated strains of highly pathogenic Eimeria rabbits and low pathogenic Eimeria rabbits; Diseased rabbit coccidia are wild strains and/or early-maturing attenuated strains.

Preferably, in the rabbit coccidiosis live vaccine composition, the precocious attenuated strain of the highly pathogenic Eimeria rabbit and the number of oocysts of the low pathogenic Eimeria rabbit The ratio is 1:(2～20). Preferably it is 1:(5-20). The compounded use of the content ratio within the above range can better ensure the safety and immune protection efficacy of the vaccine at the same time.

Preferably, the low pathogenic Rabbit coccidia adopts an early-maturing attenuated strain.

Preferably, the low pathogenicity of Eimeria rabbits includes one or more of Eimeria parvum, Eimeria pierceri, and Eimeria wiertenii.

Among the numerous highly pathogenic rabbit coccidians and low pathogenic rabbit coccidiums, the present invention finds that Eimeria perforating is effective in improving the efficiency of Eimeria large, Eimeria intestinalis and Eimeria medium. The effect that constitutes the safety of the vaccine is optimal. Therefore, in the rabbit coccidiosis live vaccine composition, the precocious attenuated strains of the highly pathogenic Eimeria rabbits preferably include Eimeria large, Eimeria intestinalis and Eimeria medium An early-maturing attenuated strain of coccidia; the low pathogenicity of Eimeria rabbits preferably includes Eimeria piercing.

On the basis of satisfying the content ratio of the above components, in order to better ensure the safety and immune protection efficacy of the vaccine at the same time, preferably, the live rabbit coccidiosis vaccine composition includes the following components: a large Eimeria ball 100-3000 oocysts/head for the precocious attenuated strain of Eimeria enterica, 500-5000 oocysts/head for the early-maturing attenuated strain of Eimeria enterica, 500-6000 eggs for the medium-sized attenuated strain of Eimeria Sac/head, 1500-40,000 oocysts/head of perforated Eimeria wild strain or precocious attenuated strain.

More preferably, the rabbit coccidiosis live vaccine composition includes the following components: 500-3000 oocysts/head of the precocious attenuated strain of Eimeria maxima, and the precocious attenuated strain of Eimeria enterica 1,000-5,000 oocysts/head, 1,000-5,000 oocysts/head of Eimeria medium-sized precocious attenuated strains, 3,000-30,000 oocysts/perforated wild strains or precocious attenuated strains of Eimeria head.

In order to expand the immune protection scope of the vaccine, the rabbit coccidiosis live vaccine composition of the present invention can contain a variety of highly pathogenic coccidia and low pathogenic coccidia, thereby forming a polyvalent coccidiosis vaccine.

As a preferred solution of the present invention, the present invention provides the precocious attenuated strains of Eimeria maxima, Eimeria intestinalis and Eimeria medium and the precocious attenuated strains of Eimeria perforating Composition of quadrivalent live coccidia vaccine composition. The immunogen composition of the rabbit coccidiosis live vaccine composition is as follows: 1000-3000 oocysts/head of the precocious attenuated strain of Eimeria maxima, 1000-4000 eggs of the precocious attenuated strain of Eimeria enterica Cyst/head portion, 2,000-5,000 oocysts/head portion of the precocious attenuated strain of medium-sized Eimeria, 3,000-30,000 oocysts/head portion of the wild strain or precocious attenuated strain of Eimeria perforating. Wherein, the ratio of the number of oocysts of Eimeria perforating to Eimeria large, Eimeria intestinalis and Eimeria medium is preferably (5-20):1.

The immunization mode of the live rabbit coccidiosis vaccine of the present invention is as follows: 20-50-day-old rabbits are orally inoculated.

Further, the present invention provides a preparation method of the rabbit coccidiosis live vaccine composition, which comprises the breeding of early-maturing attenuating worm strains; The breeding method of the steps: Orally inoculate rabbits with the coccidia strain to be attenuated, and inoculate the coccidia strain to be attenuated one or more times 12 to 72 hours after the inoculation, and harvest the earliest oocysts. Carry out passage expansion.

Preferably, the time interval between re-inoculation after the first inoculation and each re-inoculation is 12-60 hours; the high dose is 5-10 times the immunization dose of the first inoculation.

Specifically, the re-inoculation with the high dose of the coccidia strain to be attenuated once or for multiple times is re-inoculated twice at 12-72 hours and 24-84 hours after the first inoculation, respectively.

Preferably, the method for selecting and breeding the early-maturing attenuated strains comprises: inoculating the rabbits orally with the coccidia strains to be attenuated at an inoculation dose of 1.0×10 ⁵ to 2.0×10 ⁵ sporulated oocysts/rabbit, and after the first inoculation 12-72 hours after the first inoculation, the coccidia strain to be attenuated was re-inoculated orally with the inoculation dose of 5.0×10 ⁵ to 1.0×10 ⁶ sporulated oocysts/rabbit; 24-84 hours after the first inoculation, the attenuated balls were Insect strains were orally inoculated once with an inoculation dose of 1.0×10 ⁶ to 2.0×10 ⁶ sporulated oocysts/rabbit, and the earliest excreted oocysts were harvested for passage expansion.

The present invention also provides the application of the live rabbit coccidiosis vaccine composition or the live rabbit coccidiosis vaccine composition obtained by the preparation method in preparing a preparation for preventing or treating rabbit coccidiosis.

The present invention provides a rabbit coccidiosis live vaccine, which comprises the rabbit coccidiosis live vaccine composition.

Preferably, the live rabbit coccidiosis vaccine further comprises adjuvants and adjuvants permitted in the field of animal vaccines.

In another aspect, the present invention provides a method for breeding coccidia early maturing attenuated strains. Coccidia strains were orally inoculated one or more times, and the earliest excreted oocysts were harvested for passage expansion.

Preferably, the time interval between re-inoculation after the first inoculation and each re-inoculation is 12-60 hours; the high dose is 5-10 times the immunization dose of the first inoculation. More preferably, the time interval for re-inoculation after the first inoculation is 12-48 hours, and the time interval between each re-inoculation is 12-24 hours.

The rapid weakening breeding method of the rabbit coccidia provided by the present invention is different from the classical precocious pressure breeding method that is most commonly used at present, by adding one or more high-dose inoculations, and then collecting the earliest excreted oocysts for passage The period of the early-maturing attenuated strains obtained by amplification is shortened by 2 to 5 generations compared with the classical early-maturing pressure breeding method, and the obtained early-maturing strains have stable genetic characteristics, significantly reduced pathogenicity and good immunogenicity.

Preferably, the re-inoculation with a high dose of the coccidia strain to be attenuated for one or more times is re-inoculation twice at 12-72 hours and 24-84 hours after the first inoculation, respectively. More preferably, the re-inoculation is performed twice at 12 to 48 hours and 24 to 72 hours after the first inoculation, respectively.

More preferably, the breeding method comprises: orally inoculating rabbits with the coccidia strain to be attenuated at an inoculation dose of 1.0×10 ⁵ to 2.0×10 ⁵ sporulated oocysts/rabbit, 12 to 72 hours after the first inoculation Orally inoculate the to-be-attenuated coccidial strain with an inoculation dose of 5.0×10 ⁵ to 1.0×10 ⁶ sporulated oocysts/rabbit; then 24-84 hours after the first inoculation The inoculation dose of 1.0×10 ⁶ to 2.0×10 ⁶ sporulated oocysts/rabbit was orally inoculated once, and the earliest excreted oocysts were harvested for passage expansion.

The present invention further provides the application of the method for breeding the coccidial early-maturing attenuating strains in the breeding of attenuating coccidial strains or the preparation of coccidiosis vaccines.

The present invention also provides a live coccidiosis vaccine, which comprises a coccidia early-maturing attenuated strain obtained by using the method for selecting the coccidia precocious attenuated strain.

The beneficial effects of the present invention are:

The rabbit coccidiosis live vaccine provided by the invention has high immune protection efficacy and safety at the same time, can effectively prevent and treat rabbit coccidiosis, has a long immune duration, can effectively reduce or eliminate the use of anti-coccidiosis drugs, and reduce breeding costs At the same time, the adverse effects of vaccination on the growth and production performance of rabbits such as weight gain are significantly reduced, and the breeding efficiency is improved, and the vaccine is easy to produce, has no pollution to animal products and the environment, and has good economic benefits and broad application prospects.

The method for breeding coccidia early-maturing attenuated strains provided by the invention can effectively shorten the breeding cycle of the early-maturing strains, effectively improve the breeding efficiency of the coccidia early-maturing attenuated strains, and the obtained early-maturing strains have stable genetic characteristics and pathogenicity. The performance is significantly reduced while maintaining good immunogenicity, which provides an efficient method for the selection of coccidial early-maturing attenuated strains and the preparation of coccidial vaccines.

Detailed ways

The preferred embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that the following examples are given for illustrative purposes only, and are not intended to limit the scope of the present invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from the spirit and spirit of the present invention.

The experimental methods used in the following examples are conventional methods unless otherwise specified.

The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

Example 1 Breeding method of coccidia early-maturing attenuated strains (1)

The present embodiment provides a method for breeding early-maturing attenuated strains of coccidia, comprising the following steps:

(1) First inoculation: Orally inoculate clean-grade (no coccidial infection) New Zealand white rabbits with the coccidia strain to be weakened at an inoculation dose of 1.0×10 ⁵ sporulated oocysts/rabbit;

(2) Repeated inoculation: 12 hours after the first inoculation, the coccidia strain to be weakened was orally inoculated once again with the inoculation dose of 5.0×10 ⁵ sporulated oocysts/rabbit; 24 hours after the first inoculation, the to-be-weakened The coccidia strain of 1.0×10 ⁶ sporulated oocysts/rabbit was orally inoculated once;

(3) The earliest excreted oocysts were harvested, and the sporulation was completed under suitable conditions for passage expansion.

Example 2 Breeding method of coccidia precocious attenuated strains (2)

The present embodiment provides a method for breeding early-maturing attenuated strains of coccidia, comprising the following steps:

(1) First inoculation: Orally inoculate clean-grade (no coccidial infection) New Zealand white rabbits with the coccidia strain to be weakened at an inoculation dose of 1.0×10 ⁵ sporulated oocysts/rabbit;

(2) Repeated inoculation: 24 hours after the first inoculation, the coccidial strain to be weakened was re-inoculated orally with the inoculation dose of 5.0×10 ⁵ sporulated oocysts/rabbit; 36 hours after the first inoculation, the to-be-weakened The coccidia strain of 1.0×10 ⁶ sporulated oocysts/rabbit was orally inoculated once;

(3) The earliest excreted oocysts were harvested, and the sporulation was completed under suitable conditions for passage expansion.

Example 3 Breeding method of coccidia early-maturing attenuated strains (3)

The present embodiment provides a method for breeding early-maturing attenuated strains of coccidia, comprising the following steps:

(1) First inoculation: Orally inoculate clean-grade (no coccidial infection) New Zealand white rabbits with the coccidia strain to be weakened at an inoculation dose of 2.0×10 ⁵ sporulated oocysts/rabbit;

(2) Repeated inoculation: 12 hours after the first inoculation, the coccidial strain to be weakened was re-inoculated orally with an inoculation dose of 1.0×10 ⁶ sporulated oocysts/rabbit; 24 hours after the first inoculation, the to-be-weakened The coccidia strain of 2.0×10 ⁶ sporulated oocysts/rabbit was orally inoculated once;

(3) The earliest excreted oocysts were harvested, and the sporulation was completed under suitable conditions for passage expansion.

Example 4 Breeding method of early maturing and attenuated strains of coccidia (4)

The present embodiment provides a method for breeding early-maturing attenuated strains of coccidia, comprising the following steps:

(1) First inoculation: Orally inoculate clean-grade (no coccidial infection) New Zealand white rabbits with the coccidia strain to be weakened at an inoculation dose of 2.0×10 ⁵ sporulated oocysts/rabbit;

(2) Repeated inoculation: 36 hours after the first inoculation, the coccidial strain to be weakened was re-inoculated orally with an inoculation dose of 1.0×10 ⁶ sporulated oocysts/rabbit; 48 hours after the first inoculation, the to-be-weakened The coccidia strain of 2.0×10 ⁶ sporulated oocysts/rabbit was orally inoculated once;

(3) The earliest excreted oocysts were harvested, and the sporulation was completed under suitable conditions for passage expansion.

Example 5 Breeding method of coccidia early-maturing attenuated strains (5)

The present embodiment provides a method for breeding early-maturing attenuated strains of coccidia, comprising the following steps:

(1) First inoculation: Orally inoculate clean-grade (no coccidial infection) New Zealand white rabbits with the coccidia strain to be weakened at an inoculation dose of 2.0×10 ⁵ sporulated oocysts/rabbit;

(2) Repeated inoculation: 48 hours after the first inoculation, the to-be-weakened coccidial strain was re-inoculated orally with an inoculation dose of 1.0×10 ⁶ sporulated oocysts/rabbit; 72 hours after the first inoculation, the to-be-weakened The coccidia strain of 2.0×10 ⁶ sporulated oocysts/rabbit was orally inoculated once;

(3) The earliest excreted oocysts were harvested, and the sporulation was completed under suitable conditions for passage expansion.

Experimental Example 6 Breeding of early-maturing attenuated strains

Utilize the breeding method of the coccidia early-maturing attenuated strains of Examples 1 to 5 to select and breed Eimeria large, Eimeria medium, and Eimeria intestinalis with high prevalence and strong pathogenicity. Coccidia and early-maturing attenuated strains of the less pathogenic species Eimeria perfringens. The selection and breeding method of coccidian early-maturing attenuated strains provided in Examples 1-5 and the classic early-maturity selection and breeding method reported in the literature (the classic early-maturity selection and breeding method is specifically: oral administration of a high dose of the coccidia strains to be attenuated). Inoculate and harvest the earliest excreted oocysts. For specific methods and experimental results, please refer to the literature, Jeffers T K, Attenuation ofeimeria tenella through selection for precociousness. J Parasitol, 1975, 61(6): 1083-1090) The number of breeding passages and latent The results are compared with each other, and the results are shown in Table 1. The results show that the breeding methods for coccidia early-maturing attenuated strains provided in Examples 1 to 5 require significantly fewer breeding generations, and the early-maturing strains with weakened virulence can be obtained more quickly, and the obtained strains are free of The pressure was continuously propagated for 10 generations, and the shortened latent period was still maintained (Table 1), indicating that the genetic characteristics of the attenuated insect strain obtained by the method of the present example were stable.

Table 1 Comparison of the efficiency of obtaining early-maturing attenuated strains of Coccidia by different breeding methods

Example 7 Development of a trivalent live rabbit coccidiosis vaccine by utilizing the early-maturing attenuated strains of rabbit coccidiosis

In this example, the early-maturing attenuated strains of Eimeria large, Eimeria medium and Eimeria enterica obtained by the breeding method of Example 1 are used for the preparation of live coccidiosis vaccines in rabbits. Preparation and development of a live trivalent vaccine against coccidiosis in rabbits.

First, the optimal immunization dose of each precocious attenuated strain was determined based on the effective resistance to the infection of 5.0×10 ⁴ homosporulated oocysts of the wild virus strain (Table 2). According to the experimental results shown in Table 2, the optimal immunization doses of the precocious attenuated strains are: Eimeria large 1500/rabbit, Eimeria medium 3000/rabbit and Eimeria intestinalis 2000/ rabbit.

Table 2 Screening of optimal immune doses of Eimeria large, Eimeria medium and Eimeria enterica precocious attenuated strains

Further based on the above-determined optimal immune dose of a single worm strain, carry out multi-party proportioning to determine the appropriate immunization dose of each worm strain in the trivalent vaccine, namely large Eimeria 1500/rabbit, medium Eimeria 3000 /rabbit and Eimeria enterica 2000/rabbit, according to the determined immunization dose, the trivalent rabbit coccidia live vaccine was prepared.

The prepared trivalent rabbit coccidia live vaccine was analyzed for safety and immune protection efficacy, and the specific method was as follows: after carrying out the proportioning vaccine according to the optimal immune dose determined above, orally inoculate 35-day-old New Zealand white rabbits (10 per group). only), set up immunization group, repeated immunization group, high-dose immunization group, non-immunized control group and non-immunized challenge group. The repeated immunization group was immunized again at the same dose 14 days after the optimal immunization dose; the high-dose immunization group was immunized at 10 times the recommended dose. After immunization, all rabbits were raised normally, and sufficient feed and drinking water were guaranteed. Safety evaluation: The rabbits were weighed on the 14th day after immunization, and the changes of body weight gain of the rabbits before and after immunization in each group were calculated. Evaluation of immunization efficacy: 14 days after immunization, high-dose mixed wild virus strains (each containing 1.0×10 ⁵ sporulated oocysts) were challenged, and the oocyst discharge and body weight gain after challenge were counted.

Table 3 Safety and immune protection efficacy of trivalent rabbit coccidia vaccine

The results are shown in Table 3. The results show that after the three kinds of rabbit coccidium precoccidia attenuated strains were immunized according to the optimal ratio, they had good immune protection against the infection of large doses of wild toxin strains. Immunization has a certain adverse effect on the weight gain of rabbits.

The development of embodiment 8 tetravalent rabbit coccidia live vaccine

This example further introduces the utilization example of high dose (3000 oocysts/head portion, 10,000 oocysts/head portion or 30,000 oocysts/head portion) on the basis of the trivalent coccidial live vaccine of Example 2 The early-maturing attenuated strain of Eimeria perfringens obtained by the breeding method of 1 was used to prepare a live quadrivalent rabbit coccidiosis vaccine.

The immune protection efficacy and safety of the quadrivalent rabbit coccidia live vaccine obtained above were evaluated, and the effects of the introduction of Eimeria perforating on Eimeria large, Eimeria medium and Eimeria intestinalis were analyzed. The impact on the immune efficacy and safety of worms, the specific methods are as follows:

After formulating the vaccine according to the optimal immunization dose, in the vaccine components, according to different high doses of Eimeria pieris (3000 oocysts/head, 10,000 oocysts/head or 30,000 oocysts/head) 35-day-old New Zealand white rabbits (10 per group) were orally inoculated, and the groups were grouped as shown in Table 4. After immunization, all rabbits were raised normally, and sufficient feed and drinking water were guaranteed. Safety evaluation: The rabbits were weighed on the 14th day after immunization, and the changes of body weight gain of the rabbits before and after immunization in each group were calculated. Evaluation of immunization efficacy: 14 days after immunization, high-dose mixed wild virus strains (each containing 1.0×10 ⁵ sporulated oocysts) were challenged, and the oocyst discharge and body weight gain after challenge were counted.

The results, shown in Table 4, show that the addition of high doses of Eimeria piercing is effective against a trivalent coccidial vaccine consisting of Eimeria large, Eimeria medium and Eimeria enterica. There was no significant effect on immunoprotective efficacy, but the adverse effect on body weight gain in rabbits was significantly reduced, as demonstrated by Eimeria large, Eimeria medium, Eimeria enterica, and Eimeria perfringens The live quadrivalent rabbit coccidia vaccine has both high immune protection efficacy and high safety.

Table 4 Immune efficacy and safety analysis of tetravalent rabbit coccidia live vaccine

Although the present invention has been described in detail above with general description and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention fall within the scope of the claimed protection of the present invention.

Claims (10)

1. A rabbit coccidiosis live vaccine composition, which comprises at least 3 precocious attenuated strains of highly pathogenic rabbit eimeria and low pathogenic rabbit eimeria; the low-pathogenicity rabbit Eimeria tenella is a wild worm strain and/or a precocious attenuated worm strain.

2. The live rabbit coccidiosis vaccine composition of claim 1, wherein the oocyst numbers of the highly pathogenic rabbit eimeria praecox strain and the less pathogenic rabbit eimeria coccidiosis in the live rabbit coccidiosis vaccine composition are in a ratio of 1: (2-20).

3. The rabbit coccidiosis live vaccine composition of claim 1 or 2, wherein the low pathogenic rabbit eimeria comprises one or more of eimeria minutiae, eimeria perforata, eimeria vimentis;

preferably, the precocious attenuated strains of highly pathogenic rabbit eimeria comprise precocious attenuated strains of eimeria macrogola, eimeria intestinalis and eimeria intermedia; the low pathogenic E.rabbit comprises perforated E..

4. The rabbit coccidiosis live vaccine composition according to any one of claims 1 to 3, which comprises the following components: 100-3000 oocysts per head part of precocious attenuated strain of large Eimeria, 500-5000 oocysts per head part of precocious attenuated strain of intestinal Eimeria, 500-6000 oocysts per head part of precocious attenuated strain of medium Eimeria, and 1500-40000 oocysts per head part of wild Eimeria strain or precocious attenuated strain of perforated Eimeria.

5. The rabbit coccidiosis live vaccine composition according to any one of claims 1 to 4, which comprises the following components: 500-3000 oocysts per head part of precocious attenuated strain of large Eimeria, 1000-5000 oocysts per head part of precocious attenuated strain of intestinal Eimeria, 1000-5000 oocysts per head part of precocious attenuated strain of medium Eimeria, and 3000-30000 oocysts per head part of wild Eimeria strain or precocious attenuated strain of perforated Eimeria.

6. The method for preparing the rabbit coccidiosis live vaccine composition of any one of claims 1 to 5, which comprises the breeding of a prematurity attenuated strain; the breeding of the early-maturing attenuated insect plants adopts a classical early-maturing pressure breeding method or a breeding method comprising the following steps: orally inoculating the rabbit to be attenuated coccidia strain, and orally inoculating the rabbit with the high-dose coccidia strain to be attenuated once or more for 12-72 hours after the first inoculation, and harvesting oocysts discharged earliest for passage amplification;

preferably, the first inoculation is followed by the second inoculation, and the time interval between the second inoculations is 12-60 hours; the high dose is 5-10 times of the immunization dose of the first inoculation.

7. The preparation method of claim 6, wherein the re-oral inoculation once or more times with the high-dose coccidia strain to be attenuated is re-inoculation twice within 12-72 hours and 24-84 hours after the first inoculation respectively;

preferably, the breeding method of the early-maturing attenuated insect strain comprises the following steps: the coccidia to be attenuated is added at 1.0X 10⁵～2.0×10⁵Inoculating dose of sporulated oocysts/rabbits to the rabbits orally, and inoculating the to-be-attenuated coccidian strains at 5.0 × 10 hours after the first inoculation⁵～1.0×10⁶The sporulated oocysts/rabbits were inoculated once more orally; then the first inoculation is carried out 24-84 hours later, the coccidia strain to be attenuated is treated at the ratio of 1.0 multiplied by 10⁶～2.0×10⁶The sporulated oocysts/rabbit were inoculated once orally at the inoculation dose and the earliest shed oocysts were harvested for passage expansion.

8. Use of the rabbit coccidiosis live vaccine composition of any one of claims 1-5 or the rabbit coccidiosis live vaccine composition prepared by the preparation method of claim 6 or 7 in preparation of a preparation for preventing or treating rabbit coccidiosis.

9. A breeding method of a premature and weak coccidia strain is characterized by comprising the following steps: orally inoculating the rabbit to be attenuated coccidia strain, and orally inoculating the rabbit with the high-dose coccidia strain to be attenuated once or more for 12-72 hours after the first inoculation, and harvesting oocysts discharged earliest for passage amplification;

preferably, the first inoculation is followed by the second inoculation, and the time interval between the second inoculations is 12-60 hours; the high dose is 5-10 times of the immunization dose of the first inoculation.

10. The breeding method of the coccidia precocious attenuated insect strain as claimed in claim 9, wherein the re-oral inoculation with high dose of the to-be attenuated coccidia strain is one or more times of re-inoculation for 12-72 hours and 24-84 hours after the first inoculation;

preferably, the breeding method comprises: the coccidia to be attenuated is added at 1.0X 10⁵～2.0×10⁵The sporulated oocyst/rabbit inoculation dose orally inoculates rabbits 12-7 after the first inoculationThe coccidia to be attenuated strain is treated at 5.0X 10 in 2 hours⁵～1.0×10⁶The sporulated oocysts/rabbits were inoculated once more orally; then the first inoculation is carried out 24-84 hours later, the coccidia strain to be attenuated is treated at the ratio of 1.0 multiplied by 10⁶～2.0×10⁶The sporulated oocysts/rabbit were inoculated once orally at the inoculation dose and the earliest shed oocysts were harvested for passage expansion.