Method of quickly producing antibodies against avian influenza and maintain antibody titer of duck

Abstract

The present invention provides a method for enabling ducklings to quickly produce anti-avian influenza antibodies and maintain antibody titer, including: step 1) the first immunization is performed on ducklings at the age of 5 to 15 days: each duckling is inoculated with an avian influenza inactivated antigen in an abdomen, and simultaneously intramuscularly or subcutaneously immunized with an avian influenza inactivated oil-emulsion vaccine. In the method, through the double effects of intraperitoneal inoculation with the inactivated antigen and intramuscular or subcutaneous injection with the inactivated oil-emulsion vaccine on the ducklings, the ducklings can quickly produce an immune response so as to quickly produce the anti-avian influenza antibodies; and the immune dead time of the ducklings immunized with the avian influenza inactivated vaccine can be effectively reduced by more than 7 days.

Claims

1. A method for enabling ducklings to quickly produce anti-avian influenza antibodies and maintain antibody titer, comprising: step 1) performing first immunization on ducklings at the age of 5 to 15 days: inoculating each duckling with an avian influenza inactivated antigen in an abdomen, and simultaneously intramuscularly or subcutaneously immunizing each duckling with an avian influenza inactivated oil-emulsion vaccine.

2. The method according to claim 1, wherein in step 1), the first immunization is performed on the ducklings at the age of 8 to 10 days.

3. The method according to claim 1, wherein, in step 1), the dosage of the inactivated antigens is 0.1 to 1 mL, and an HA valence is 7 to 12 Log 2.

4. The method according to claim 1, wherein, in step 1), the dosage of the inactivated antigens is 0.3 to 0.5 mL, and the HA valence is 9 to 10 Log 2.

5. The method according to claim 1, wherein, in step 1), the dosage of the inactivated oil-emulsion vaccines is 0.1 to 1 mL, and an HI valence is 7 to 12 Log 2.

6. The method according to claim 1, wherein, in step 1), the dosage of the inactivated oil-emulsion vaccines is 0.3 to 0.5 mL, and the HI valence is 9 to 10 Log 2.

7. The method according to claim 1, further comprising step 2) re-immunizing the ducklings after 10 days from the first immunization: inoculating each duckling with the avian influenza inactivated antigen in an abdomen.

8. The method according to claim 7, wherein in step 2), the dosage of the inactivated antigens is 0.1 to 1 mL, and the HA valence is 7 to 12 Log 2.

9. The method according to claim 7, wherein, in step 2), the dosage of the inactivated antigens is 0.3 to 0.5 mL, and the HA valence is 9 to 10 Log 2.

10. The method according to claim 1, wherein in the step 1), strains of the inactivated antigens comprise an H5 subtype avian influenza strain and an H9 subtype avian influenza strain; and a production strain of the inactivated oil-emulsion vaccine comprises an H5 subtype avian influenza strain and an H9 subtype avian influenza strain.

Description

DETAILED DESCRIPTION

[0022] The present invention provides a method for enabling ducklings to quickly produce anti-avian influenza antibodies and maintain antibody titer, including: step 1) immunizing ducklings at the age of 5 to 15 days: each duckling is inoculated with an avian influenza inactivated antigen in an abdomen, and simultaneously intramuscularly or subcutaneously immunizing the ducklings with avian influenza inactivated oil-emulsion vaccines.

[0023] In the following specific embodiments, unless otherwise specified, the employed reagents or test objects are commercially available.

[0024] In the following specific embodiments, the employed ducklings are commercially-available local ducklings in Guangdong Province, inactivated antigens and inactivated oil-emulsion vaccines are from Guangdong Wens Dahuanong Biotechnology Co. Ltd., and an inactivated antigen strain includes an H5 subtype avian influenza strain and an H9 subtype avian influenza strain, such as H5 Re-6 strain and H9 SS strain; and the inactivated oil-emulsion vaccine is H5+H9 subtype avian influenza inactivated vaccine, and the production strain thereof includes an H5 subtype avian influenza strain and H9 subtype avian influenza strain, such as H5 Re-6 strain and H9 SS strain.

Embodiment 1

[0025] 50 commercially-available newly hatched ducklings are obtained and divided into six experimental groups, wherein group 1 is a control group and is not immunized; group 2 receives first immunization at the age of one day; group 3 receives the first immunization at the age of 5 days; group 4 receives the first immunization at the age of 8 days; group 5 receives the first immunization at the age of 10 days; group 6 receives the first immunization at the age of 14 days; and groups 2 to 6 are re-immunized after 10 days from the first immunization.

[0026] For the ducklings in the experimental groups 1 to 6, blood is collected from veins under duck wings on 0, 5th, 7th, 14th, 21st, 28th, 35th and 42nd days after the first immunization, serum is isolated from the blood, and the valences (HI valences) of anti-avian influenza virus antibodies in the serum are uniformly determined Results are shown in Table 1.

[0027] In the present embodiment, the first immunization specifically includes the following operation steps: each duckling is injected with 0.5 mL of avian influenza inactivated antigen in the abdomen, and simultaneously is subcutaneously injected with 0.5 mL of avian influenza inactivated oil-emulsion vaccine in the neck.

[0028] In the present embodiment the re-immunization treatment specifically includes the following operation step: each duckling is injected with 0.5 mL of avian influenza inactivated antigen in the abdomen.

TABLE-US-00001 TABLE 1 Impact of Composite Immunization of Avian Influenza Inactivated Antigens and Inactivated Vaccines on Avian Influenza Antibodies of Ducklings Monitoring Time and Experi- Antibody Titer (HI Valence) mental Vac- 0 5th 7th 14th 21st 28th 35th 42nd group cine day day day day day day day day Group 1 H5 6 5 4 2 0 0 0 0 H9 4 3 2 1 0 0 0 0 Group 2 H5 6 5 4 2 3 4 4 3 H9 4 3 4 3 4 4 4 3 Group 3 H5 2 5 5 6 6 7 5 4 H9 1 4 6 6 7 7 7 7 Group 4 H5 3 4 5 7 8 7 6 5 H9 2 6 7 8 9 9 8 8 Group 5 H5 2 5 6 7 8 8 8 7 H9 1 6 7 8 9 9 8 8 Group 6 H5 2 5 6 7 8 8 8 7 H9 1 6 8 8 9 9 9 8

[0029] The results show that after the ducklings are injected with the immune avian influenza inactivated antigens, in the abdomens and subcutaneously injected with the inactivated oil-emulsion vaccines, after 5 days, the H5 subtype avian influenza antibody titer can reach 4 to 5 Log 2 and the H9 subtype avian influenza antibody titer can reach 3 to 6 Log 2; for the ducklings receiving the first immunization at the age of more than 5 days, after 7 days, the H5 subtype avian influenza antibody titer can reach 5 to 7 Log, the H9 subtype avian influenza antibody titer can reach 6 to 8 Log 2, and the titer can be maintained for more than 35 days; and for the ducklings receiving the first immunization at the age of 8 to 10 days, the avian influenza H5 and H9 subtypes avian influenza antibody titers are greater than 4 Log 2 after 5 days; after 7 days, the H5 subtype avian influenza antibody titer can reach 5 to 7 Log 2, the H9 subtype avian influenza antibody titer can reach 7 to 9 Log 2, and the titer can be maintained for more than 42 days.

Embodiment 2

[0030] 50 commercially-available newly-hatched ducklings are obtained and divided into 5 experimental groups, wherein group 1 is a control group and is not immunized.

[0031] Groups 2 to 5 receive the first immunization at the age of 15 days, wherein each duckling in group 2 receives the first immunization by means of intramuscular injection with 0.5 mL of avian influenza inactivated antigen; each duckling in group 3 receives the first immunization by means of subcutaneous injection with 0.5 mL of avian influenza inactivated antigen in the neck; each duckling in group 4 receives the first immunization by means of intranasal immunization with 0.5 mL of avian influenza inactivated antigen; and each duckling in group 5 receives the first immunization by means of intraperitoneal injection with 0.5 mL of avian influenza inactivated antigen.

[0032] Groups 2 to 5 receive re-immunization after 10 days from the first immunization, and the re-immunization includes the following specific operation steps:

[0033] Blood is collected from veins under wings of the ducklings on 0, 5th, 7th, 14th, 21st, 28th, 35th and 42nd days after the first immunization, serum is isolated, the valences (HI valences) of anti-avian influenza virus antibodies in the serum are uniformly determined; and results are shown in Table 2.

TABLE-US-00002 TABLE 2 Effects of Different Avian Influenza Inactivated Antigen Immunization Ways on Ducklings Monitoring Time and Antibody Experi- Titer (HI Valence) mental Vac- 0 5th 7th 14th 21st 28th 35th 42nd group cine day day day day day day day day Group 1 H5 1 1 0 0 0 0 0 0 H9 2 1 0 0 0 0 0 0 Group 2 H5 1 2 3 1 0 0 0 0 H9 1.5 2 2 0 0 0 0 0 Group 3 H5 1 2 2 0 0 0 0 0 H9 2 3 3 2 1 0 0 0 Group 4 H5 1 3 2 1 0 0 0 0 H9 1 4 2 4 3 0 0 0 Group 5 H5 2 5 6.5 7 8 7 4 1 H9 1 6 7 8 9 8 5 2

[0034] It can be known from Table 2 that for the ducklings in group receiving the immunization with the intraperitoneal injection way, after 5 days from the first immunization, the H5 subtype avian influenza antibody titer can reach as high as 5 Log 2, and the H9 antibody titer can reach as high as 6 Log 2 and both the H5 subtype avian influenza antibody titer and the H9 subtype avian influenza antibody titer can be kept at more than 7 Log 2 after 28 days from the first immunization. For the ducklings receiving the immunization with the intramuscular injection way, the neck subcutaneous injection way or the intranasal immunization way, the H5 subtype avian influenza antibody titer and the H9 subtype avian influenza antibody titer are very low.

Embodiment 3

[0035] 50 commercially-available newly hatched ducklings are obtained and divided into 5 experimental groups, wherein group 1 is served as a control group and is not immunized with the avian influenza inactivated vaccine.

[0036] Groups 2 to 5 receive the first immunization at the age of 15 days, wherein each duckling in group 2 receives the intramuscular immunization with 0.1 mL of avian influenza inactivated antigen; each duckling in group 3 receives the intramuscular immunization with 0.3 mL of avian influenza inactivated antigen; each duckling in group 4 receives the intramuscular immunization with 0.5 mL of avian influenza inactivated antigen; and each duckling in group 5 receives the intramuscular immunization with of avian influenza inactivated antigen. The re-immunization is performed after 10 days from the first immunization, and the immunization dosage and the immunization way of the re-immunization are the same as those of the first immunization.

[0037] Blood is collected from veins under wings of the ducklings on 0, 5th, 7th, 14th, 21st, 28th, 35th and 42nd days after the first immunization, serum is isolated, the valences (HI valences) of anti-avian influenza virus antibodies in the serum are uniformly determined; and results are shown in Table 3.

TABLE-US-00003 TABLE 3 Effects of Intramuscular Immunization with Avian Influenza Inactivated Antigen on Ducklings Monitoring Time and Antibody Experi- Titer (HI Valence) mental Anti- 0 5th 7th 14th 21st 28th 35th 42nd group gen day day day day day day day day Group 1 H5 2 1 0.5 0 0 0 0 0 H9 3 2 1 0 0 0 0 0 Group 2 H5 1 5 6 6 5 3 2 0 H9 1.5 6 7 7 5.5 3 2 0 Group 3 H5 1 6 7 7 5.5 4 2 1 H9 2 7 8 9 6 4 3 1 Group 4 H5 2 6 8 8 7 5 3 1 H9 1 7 8 9 7 6 4 2 Group 5 H5 2 6 8 9 7 6 3 1 H9 1 7 8 9 8 7 4 2

[0038] It can be known from Table 3 that when the ducklings receive the intraperitoneal immunization with 0.1 to 1 mL of avian influenza inactivated antigen, the avian influenza H5 subtype and H9 subtype avian influenza antibody titers in a duckling body can reach more than 5 Log 2 after 5 days from the immunization; and when the ducklings receive the intraperitoneal immunization with 0.3 to 0.5 mL of avian influenza inactivated antigen, the avian influenza H5 subtype and H9 subtype avian influenza antibody titers in the duckling body, can be kept at 5 Log 2 for more than 21 days.

Embodiment 4

[0039] 50 commercially-available newly hatched ducklings are obtained and divided into 5 experimental groups, wherein group 1 is served as a control group and is not immunized with the avian influenza inactivated vaccine.

[0040] Ducklings in groups 2 to 5 receive the first immunization by means of subcutaneous injection with the avian influenza inactivated vaccine at the age of 15 days; each duckling in group 2 is immunized with 0.1 mL of avian influenza inactivated vaccine; each duckling in group 3 is immunized with 0.3 mL of avian influenza inactivated vaccine; each duckling in group 4 is immunized with 0.5 mL of avian influenza inactivated vaccine; and each duckling in group 5 is immunized with 1 mL of avian influenza inactivated vaccine.

[0041] Blood is collected from veins under wings of the ducklings on 0, 5th, 7th, 14th, 21st, 28th, 35th and 42nd days after the first immunization, serum is isolated, the valences (HI valences) of anti-avian influenza virus antibodies in the serum are uniformly determined; and results are shown in Table 4.

TABLE-US-00004 TABLE 4 Effects for Immunizing Ducklings with Avian Influenza Inactivated Vaccine Monitoring Time and Antibody Experi- Titer (HI Valence) mental Vac- 0 5th 7th 14th 21st 28th 35th 42nd group cine day day day day day day day day Group 1 H5 2 2 1 1 0 0 0 0 H9 1 1 1 0 0 0 0 0 Group 2 H5 1 1 2 2 3 4 5 5 H9 1.5 2 3 4 5 6 6 6 Group 3 H5 1 2 3 4 5 6 6 6 H9 1.5 2 4 7 8.5 8 8.5 8 Group 4 H5 2 1 2 3 5 6 7 6 H9 1 1 3 6 8 8.5 9 9 Group 5 H5 0.5 1 2 3 5 6 6 6 H9 0.5 2 5 6 8 9 9 8

[0042] It can be known from Table 4 that after 21 days from that the ducklings are immunized with the avian influenza inactivated vaccine, the H5 subtype avian influenza specific antibody-valence (HI valence) is 3 to 5 Log 2 and the H9 subtype avian influenza specific antibody valence (HI valence) is 5 to 8 Log 2; and the avian influenza specific antibody valences are relatively low;after 0 to 14 days from the immunization and insufficient to play a role in protection.

[0043] The above embodiments are only preferred embodiments of the present invention and may not be used to limit the protection scope of the present invention. Any insubstantial variations and replacements made by those skilled in the art based on the present invention shall fall within the protection scope claimed by the present invention.