RABBIT COCCIDIOSIS VACCINE AND APPLICATION THEREOF

Abstract

The present invention discloses a rabbit coccidiosis vaccine and application thereof, the vaccine comprises 100 to 800 Eimeria media/dose, 200 to 1600 Eimeria magna/dose, and 100 to 800 Eimeria intestinalis/dose. The composition of the vaccine possesses the characteristics as scientific reasonable, low cost, no drug residue or drug resistance or environmental pollution would occur, good immunogenicity and safe to use. After oral immunization in rabbits, it may effectively resist the infections of 110.sup.5 Eimeria media, 510.sup.4 Eimeria magna and 310.sup.3 Eimeria intestinalis. It also may be used to prepare a pharmaceutical preparation against rabbit coccidiosis.

Claims

1-6. (canceled)

7. A rabbit coccidiosis vaccine, wherein the rabbit coccidiosis vaccine comprises: 100 to 800 Eimeria media/dose, 200 to 1600 Eimeria magna/dose, and 100 to 800 Eimeria intestinalis/dose; wherein the Eimeria media is a precocious strain of Eimeria media, the Eimeria magna is a precocious strain of Eimeria magna, and the Eimeria intestinalis is a precocious strain of Eimeria intestinalis; the rabbit coccidiosis vaccine is suitable for rabbits aged from 25 to 90 days.

8. The rabbit coccidiosis vaccine according to claim 7, wherein the vaccine comprises 300 Eimeria media/dose, 600 Eimeria magna/dose, and 600 Eimeria intestinalis/dose.

9. The rabbit coccidiosis vaccine according to claim 7, wherein the rabbit coccidiosis vaccine is suitable for oral immunization in rabbits, the oral immunization comprises mixing feed immunization, drinking water immunization and intranasal immunization.

10. A method for immunizing Eimeria in rabbits, comprising administering the rabbit coccidiosis vaccine of claim 7 to rabbits in need thereof.

11. The method according to claim 10, wherein the Eimeria in rabbits comprises Eimeria media, Eimeria magna and Eimeria intestinalis.

12. The method according to claim 10, wherein the vaccine is suitable for oral immunization in rabbits aged from 25 to 90 days.

13. The method according to claim 12, wherein the oral immunization comprises mixing feed immunization, drinking water immunization and intranasal immunization.

Description

DETAILED DESCRIPTION

[0012] The present invention provides a rabbit coccidiosis vaccine and application thereof. In order to make the objects, technical solutions and effects of the invention more clear and explicit, the present invention will be further described in detail as follows by the illustration of embodiments. It should be understood that the specific embodiments described herein merely illustrate the present invention and are not intended to limit the present invention.

[0013] The present invention provides a rabbit coccidiosis vaccine, comprising: 100 to 800 Eimeria media/dose, 200 to 1600 Eimeria magna/dose, 100 to 800 Eimeria intestinalis/dose. Eimeria media, Eimeria magna and Eimeria intestinalis are widespread and moderately or strongly toxic, and are more harmful to rabbits. The vaccine is obtained by mixing precocious strains of the above three Eimeria species that are more harmful to rabbits at a specific ratio. The obtained vaccine has good immunogenicity and is safe to use. The above dose may guarantee the immune effect with low cost, and may control the vaccine response at a low level, thereby to reduce the influence on the production of rabbits. When the values of immune doses of Eimeria media, Eimeria magna, and Eimeria intestinalis are less than the above values, it is unable to provide effective immune protection to rabbits; when the values of immune doses of Eimeria media, Eimeria magna, and Eimeria intestinalis are greater than the above values, the immune response is too strong, which will produce great influence on the production of rabbits.

[0014] Preferably, the vaccine comprises 300 Eimeria media/dose, 600 Eimeria magna/dose, and 600 Eimeria intestinalis/dose; applying the above doses may lead to the best immune effect of vaccine, low cost, slight side reactions and low influence on the production of rabbits, thus it is the best ratio.

[0015] Preferably, the Eimeria media is a precocious strain of Eimeria media, the Eimeria magna is a precocious strain of Eimeria magna, and the Eimeria intestinalis is a precocious strain of Eimeria intestinalis. The precocious strains are less pathogenic than the original strains, but may retain immunogenicity, so the immune effect of vaccines is fully guaranteed; moreover, it is safer to use and avoiding potential poisoning threats and excessive damage to the intestinal tracts of rabbits during use that may cause the harms such as the reduction of the production performance and increase of the breeding costs, etc.

[0016] In the applications of the rabbit coccidiosis vaccine, the vaccine is used for the immunization of rabbit Eimeria.

[0017] Specifically, the rabbit Eimeria comprises Eimeria media, Eimeria magna, and Eimeria intestinalis; the vaccine may effectively resist the infection of 110.sup.5 Eimeria media, 510.sup.4 Eimeria magna and 310.sup.3 Eimeria intestinalis.

[0018] Specifically, the vaccine is applied to rabbits of various strains or for different purposes, including New Zealand rabbits, Hyla rabbits, REX rabbits, California rabbits, meat rabbits, long-haired rabbits, pet rabbits, and experimental rabbits, etc.

[0019] Specifically, the vaccine is suitable for oral immunization in rabbits aged from 25 to 90 days; the vaccine is orally immunized, including mixing feed immunized, drinking water immunized or intranasal immunized, etc.

Example 1 Formulation of Rabbit Coccidiosis Vaccine

[0020] The sporulated oocysts of Eimeria media, Eimeria magna, and Eimeria intestinalis were counted by a blood counting chamber, respectively, and the concentrations were obtained. The required volume of coccidial oocysts of each component was calculated according to the ratios shown in Table 1 and placed in a clean container successively, then preservation solution was added and stirred to obtain the oocyst solution of the mixed strains.

TABLE-US-00001 TABLE 1 Ratio of components of rabbit coccidiosis vaccine. Eimeria media Eimeria magna Eimeria intestinalis (pcs/dose) (pcs/dose) (pcs/dose) Ratio 1 100 200 800 Ratio 2 800 200 100 Ratio 3 300 600 600 Ratio 4 500 500 500 Ratio 5 100 1600 100 Ratio 6 400 1200 400

Example 2

[0021] 50 normal-class rabbits aged from 25-90 days without coccidiosis tested by fecal examination were divided into 10 groups, 5 in each group. The rabbits in each group were evenly distributed according to the age in days. Among them, groups 1, 2 and 3 were immunized and challenged groups of ratio 1, and groups 4, 5 and 6 were immunized and challenged groups of ratio 2, and groups 7, 8 and 9 were non-immunized and challenged groups, and group 10 was non-immunized and non-challenged group. Rabbits in the 10 groups were separately fed under the same conditions, and the rabbits were weighed and challenged on the 14th day by orally immunized with the corresponding vaccines. Rabbits in the groups 1, 4 and 7 were orally challenged by 110.sup.5 Eimeria media sporulated oocysts/rabbit; rabbits in the groups 2, 5 and 8 were orally challenged by 510.sup.4 Eimeria magna sporulated oocysts/rabbit; and rabbits in the groups 3, 6 and 9 were orally challenged by 310.sup.3 Eimeria intestinalis sporulated oocysts/rabbit, and then observed for 12 days. The feces on the 5th and 6th days after challenge on the groups 1, 4 and 7 were collected and the number of oocysts was calculated, the feces on the 8th and 9th days after challenge on the groups 2, 5 and 8 were collected and the number of oocysts was calculated, and the feces on the 11th and 12th days after challenge on the groups 3, 6 and 9 were collected and the number of oocysts was calculated. On the 12th day after challenge, the body weights of rabbits in each group were weighed and the relative weight gain rate was calculated. The average relative weight gain rates and the relative decrease rates of OPG (oocysts per gramme) among different groups were compared.

[0022] The groups of ratio 3 and ratio 4, ratio 5 and ratio 6 as the combined immunized group were tested in two separate experiments according to the same test method described above.

TABLE-US-00002 TABLE 2 The weight gains in various groups at ratio 1 and ratio 2 Ratio of Average Average Average strains weight rate of Average Average rate of (media: Number Average Average gain relative weight weight relative magna: of weight before weight after weight after gain after weight intest- Challenged rabbits immunization/ before immunization gain challenge/ challenge gain Mortality/ Group inalis) type (n) g challenge/g a/g a/% g b/g b/% % Ratio 1: 100: media 5 1062.6 1441.3 378.7 97.2 1792.7 351.4 72 0 immunized and 200: magna 5 981.3 1341.6 360.3 92.5 1676.4 334.8 68.6 0 challenged group 800 intestinalis 5 917.5 1240.5 323 82.9 1595.3 354.8 72.7 0 Ratio 2: 800: media 5 1035.3 1359.4 324.1 83.2 1765 405.6 83.1 0 immunized and 200: magna 5 819.3 1182 362.7 93.1 1523.2 341.2 69.9 0 challenged group 100 intestinalis 5 918 1289.3 371.3 95.3 1584.1 294.8 60.4 0 Non-immunized media 5 933.9 1324.4 390.5 100.9 1565.5 241.1 49.4 0 and challenged magna 5 789.7 1178.8 389.1 99.9 1384.3 205.5 42.1 0 group intestinalis 5 821.2 1208.3 387.1 99.4 1397.2 188.9 38.7 0 Non-immunized 5 977.7 1367.3 389.6 1855.4 488.1 0 and non-challenged group

[0023] Note: In table 2, media represented Eimeria media; magna represented Eimeria magna; intestinalis represented Eimeria intestinalis; a indicated the data of weight gain on the 14th day after immunization; b indicated the data of weight gain on the 12th day after challenge.

TABLE-US-00003 TABLE 3 The weight gains in various groups at ratio 3 and ratio 4 Average Average Ratio of Average Average Average rate of Average Average rate of strains Number weight weight weight relative weight weight relative (media: Type of before before gain after weight after gain after weight magna: of rabbits immunization/ challenge/ immunization gain challenge/ challenge gain Mortality/ Group intestinalis) challenge (n) g g a/g a/% g b/g b/% % Ratio 3: 300: media 5 928.5 1317.6 389.1 93.4 1642.2 324.6 77.1 0 immunized 600: magna 5 877.0 1263.2 386.2 92.7 1549.9 286.7 68.1 0 and challenged 600 intestinalis 5 861.9 1250.6 388.7 93.3 1491 240.4 57.1 0 group Ratio 4: 500: media 5 941.2 1314.1 372.9 89.5 1641.2 327.1 77.7 0 immunized 500: magna 5 887.1 1265.4 378.3 90.8 1555.9 290.5 69.0 0 and challenged 500 intestinalis 5 963.1 1338.5 375.4 90.1 1559.9 221.4 52.6 0 group Non-immunized media 5 884.3 1295.9 411.6 98.8 1497.1 201.2 47.8 0 and challenged magna 5 969.3 1396.7 427.4 102.6 1568 171.3 40.7 0 group intestinalis 5 757.2 1171.3 414.1 99.4 1303.9 132.6 31.5 0 Non-immunized 5 906.3 1322.9 416.6 1743.9 421.0 0 and non-challenged group

[0024] Note: In table 3, media represented Eimeria media; magna represented Eimeria magna; intestinalis represented Eimeria intestinalis; a indicated the data of weight gain on the 14th day after immunization; b indicated the data of weight gain on the 12th day after challenge.

TABLE-US-00004 TABLE 4 The weight gains in various groups at ratio 5 and ratio 6 Ratio of Average Average Average Average Average strains weight Average weight rate of Average weight rate of (media: Number before weight gain after relative weight gain relative magna: of immun- before immun- weight after after weight intestinal Type of rabbits ization/ challenge/ ization gain challenge/ challenge gain Group is) challenge (n) g g a/g a/% g b/g b/% Mortality/% Ratio 5: 100: media 5 937.6 1332.6 395 92.8 1659.5 326.9 72.3 0 immunized 1600: magna 5 813.2 1155.8 342.6 80.5 1499 343.2 78 0 and 100 intestinalis 5 1002.2 1401.8 399.6 93.9 1609.5 207.7 47.2 0 challenged group Ratio 6: 400: media 5 741.2 1097 355.8 83.6 1441.1 344.1 78.2 0 immunized 1200: magna 5 959.1 1304.7 345.6 81.2 1631.6 326.9 74.3 0 and 400 intestinalis 5 1014.9 1375.8 360.9 84.8 1626.2 250.4 56.9 0 challenged group Non- media 5 984.5 1416.5 432 101.5 1629.5 213 48.4 0 immunized magna 5 1004.4 1425.3 420.9 98.9 1606.1 180.8 41.1 0 and intestinalis 5 918.3 1360.5 442.2 103.9 1500 139.5 31.7 0 challenged Non- 5 837.9 1263.5 425.6 1703.5 440 0 immunized and non- challenged group

[0025] Note: In table 4, media represented Eimeria media; magna represented Eimeria magna; intestinalis represented Eimeria intestinalis; a indicated the data of weight gain on the 14th day after immunization; b indicated the data of weight gain on the 12th day after challenge.

TABLE-US-00005 TABLE 5 Counting results of oocysts in various groups at ratio 1 and ratio 2 Ratio of strains OPG (media: value/ OPG Oocyst Relative magna: Type of piece/g decrease production/ten decrease rate of Group intestinalis) challenge feces rate/% thousand oocysts/% Ratio 1: 100: media 506000 80.9 1762.1 79.5 immunized and 200: magna 526500 54.5 2468.9 53.0 challenged 800 intestinalis 3934500 64.2 4485.1 62.2 group Ratio 2: 800: media 190750 92.8 862.7 90.0 immunized 200: magna 539150 53.4 2453.1 53.3 challenged 100 intestinalis 5550000 49.5 6134.4 48.3 group Non-immunized media 2649000 8595.5 and challenged magna 1157000 5252.9 group intestinalis 10990000 11865.3

[0026] Note: In table 5, media represented Eimeria media; magna represented Eimeria magna; intestinalis represented Eimeria intestinalis.

TABLE-US-00006 TABLE 6 Counting results of oocysts in various groups at ratio 3 and ratio 4 Ratio of strains OPG Relative (media: value/ OPG Oocyst decrease magna: Type of piece/g decrease production/ten rate of Group intestinalis) challenge feces rate/% thousand oocysts/% Ratio 3: 300: media 277550 89.8 929.9 88.5 immunized and 600: magna 483200 57.2 2632.8 52.4 challenged 600 intestinalis 5557000 56.0 4134.6 55.6 group Ratio 4: 500: media 272100 90.0 789.6 90.2 immunized 500: magna 479800 57.5 2371.5 57.1 challenged 500 intestinalis 5822500 53.9 4339.4 53.4 group Non-immunized media 2721000 8057.4 and challenged magna 1129000 5527.9 group intestinalis 12630000 9312.1

[0027] Note: In table 6, media represented Eimeria media; magna represented Eimeria magna; intestinalis represented Eimeria intestinalis.

TABLE-US-00007 TABLE 7 Counting results of oocysts in various groups at ratio 5 and ratio 6 Ratio of strains OPG Relative (media: value/ OPG Oocyst decrease magna: Type of piece/g decrease production/ten rate of Group intestinalis is) challenge feces rate/% thousand oocysts/% Ratio 5: 100: media 500300 81.8 1559.2 80.5 immunized and 1600: magna 359000 63.2 1970.7 62.4 challenged 100 intestinalis 4503100 51.0 5697.5 50.6 group Ratio 6: 400: media 244650 91.1 1839.1 77.0 immunized 1200: magna 385300 60.5 2101.8 59.9 challenged 400 intestinalis 4080300 55.6 4867.1 57.8 group Non-immunized media 2749000 7996.3 and challenged magna 975500 5241.3 group intestinalis 9190000 11533.4

[0028] Note: In table 7, media represented Eimeria media; magna represented Eimeria magna; intestinalis represented Eimeria intestinalis.

[0029] The test results were shown in Tables 2 to 7. Before the challenge, the average rates of relative weight gain of various groups immunized with the vaccines at ratios 1 to 6 were 90.7%, 90.5%, 93.1%, 90.1%, 89.1%, 83.2. %, respectively, and no death occurred in rabbits of each immunized group. After the challenge, the rates of relative weight gains in various groups challenged with Eimeria media, Eimeria magna and Eimeria intestinalis at the ratio 1 were 76.0%, 68.6%, and 72.7%, respectively; likewise, the rates of relative weight gains in various vaccine immunized groups at the ratio 2 were 83.1%, 69.9%, and 60.4%, respectively; the rates of relative weight gains in various vaccine immunized groups at the ratio 3 were 77.1%, 68.1%, and 57.1%, respectively; the rates of relative weight gains in various vaccine immunized groups at the ratio 4 were 77.7%, 69.0%, and 52.6%, respectively; the rates of relative weight gains in various vaccine immunized groups at the ratio 5 were 72.3%, 78.0%, and 47.2%, respectively; the rates of relative weight gains in various vaccine immunized groups at the ratio 6 were 78.2%, 74.3%, and 56.9%, respectively.

[0030] After challenge, the relative OPG decrease rates and the relative decrease rates of oocysts in various groups challenged with Eimeria media, Eimeria magna and Eimeria intestinalis at the ratio 1 were 80.9%, 54.5%, 64.2% and 79.5%, 53.0%, 62.2%, respectively; those in various groups at the ratio 2 were 92.8%, 53.4%, 49.5% and 90.0%, 53.3%, 48.3%, respectively; those in various groups at the ratio 3 were 89.8%, 57.2%, 56.0% and 88.5%, 52.4%, 55.6%, respectively; those in various groups at the ratio 4 were 90.0%, 57.5%, 53.9% and 90.2%, 57.1%, 53.4%, respectively; those in various groups at the ratio 5 were 81.8%, 63.2%, 51.0%, and 80.5%, 62.4%, and 50.6%, respectively; those in various groups at the ratio 6 were 91.1%, 60.5%, 55.6%, and 77.0%, 59.9%, and 57.8%, respectively.

[0031] Apparently, the rate of relative weight gains in each group immunized with the vaccines at ratios 1 to 6 was significantly higher than that in the corresponding non-immunized and challenged group, specifically, the rate of relative weight gains at the ratio 1 was higher than that in the corresponding non-immunized and challenged group by 22.6% to 34.0%; the rate of relative weight gains at the ratio 2 was higher than that in the corresponding non-immunized and challenged group by 21.7% to 33.7%; the rate of relative weight gains at the ratio 3 was higher than that in the corresponding non-immunized and challenged group by 25.6% to 29.3%; the rate of relative weight gains at the ratio 4 was higher than that in the corresponding non-immunized and challenged group by 21.1% to 29.9%; the rate of relative weight gains at the ratio 5 was higher than that in the corresponding non-immunized and challenged group by 15.5% to 36.9%; the rate of relative weight gains at the ratio 6 was higher than that in the corresponding non-immunized and challenged group by 25.2% to 36.9%. Moreover, the relative OPG decrease rates and the relative decrease rates of oocysts in various immunized and challenged groups after challenged with Eimeria media, Eimeria magna and Eimeria intestinalis were higher than 77.0% 52.4% and 48.3% respectively, indicating that various groups at the ratios 1 to 6 achieved a good immune effect and had a low effect on the weight gain of rabbits, and it was effective to resist the challenges of Eimeria media, Eimeria magna and Eimeria intestinalis at high doses.

[0032] Further, before challenge, the rate of relative weight gains in the group immunized with the vaccines at ratio 3 was significantly superior to that in the groups immunized with vaccines at other five ratios; after challenge, the rates of relative weight gains and the relative OPG decrease rates in the groups immunized with the vaccines at ratios 3, 4 and 6 were superior to those in the groups immunized with the vaccines at ratios 1, 2 and 5. Thus, by combining the influence of vaccines on the weight gains of rabbits during immunization with the relative OPG decrease rates and the relative decrease rates of oocysts after challenge, the applicant concluded that the optimal ratio of the vaccine was the ratio 3, that is, when each dose of vaccine contained 300 sporulated oocysts of Eimeria media precocious strain, 600 sporulated oocysts of Eimeria magna precocious strain and 600 sporulated oocysts of Eimeria intestinalis precocious strain, it had the slightest effect on the rabbit's body during the immunization and provided relatively strong protection effect of immunization after challenge.

[0033] The test results of rabbits in other ages in days also showed that vaccines with ratios 1 to 6 were safe to use, and the ratio 3 was the optimal ratio.

[0034] It should be understood that those skilled in the art may make equivalent substitutions or changes to the technical solutions and concepts according to the present invention, and all such changes or substitutions shall fall into the scope of protection of the appended claims.