A COMBINATION OF VACCINES TO PROPHYLACTICALLY TREAT A PIG

Abstract

The invention pertains to a combination of a first vaccine comprising anon-replicating immunogen of porcine circovirus type 2 (PCV-2) and a non-replicating immunogen of Mycoplasma hyopneumoniae, and a second vaccine comprising a live attenuated porcine reproductive and respiratory syndrome (PRRS) virus, for use in prophylactically treating a pig against an infection with PCV-2, an infection with Mycoplasma hyopneumoniae and an infection with PRRS virus, by associated separate injection of the first vaccine and the second vaccine into a tissue of the pig at a first and a second injection site respectively, wherein the first and second injection sites are at most 5 cm apart from each other.

Claims

1. (canceled)

2. The method of prophylactically treating of claim 12, wherein the first and second injection sites are at most 4 cm apart from each other.

3. The method of prophylactically treating of claim 2, wherein the first and second injection sites are at most 3 cm apart from each other.

4. The method of prophylactically treating of claim 12, wherein the associated separate injection of the first vaccine and the second vaccine occur simultaneously.

5. The method of prophylactically treating of claim 12, wherein the first and second vaccine through said injection are deposited at least partly into muscular tissue of the pig.

6. The method of prophylactically treating of claim 5, wherein the first and second vaccine are injected either by a hypodermic syringe that extends into the muscular tissue or by a jet stream of the respective vaccines using a needle-less device, wherein the jet stream penetrates the skin of the pig.

7. The method of prophylactically treating of claim 12, wherein the first vaccine and the second vaccine are administered by a single dose.

8. The method of prophylactically treating of claim 12, wherein the non-replicating immunogen of PCV2 is recombinantly expressed protein encoded by the ORF2 gene of PCV2.

9. The method of prophylactically treating of claim 8, wherein the non-replicating immunogen of PCV2 is baculovirus expressed protein of PCV2.

10. The method of prophylactically treating of claim 12, wherein the non-replicating immunogen of Mycoplasma hyopneumoniae is a Mycoplasma hyopneumoniae bacterin.

11. A kit-of-parts for use in prophylactically treating a pig against an infection with porcine circo virus type 2, an infection with Mycoplasma hyopneumoniae and an infection with PRRS virus, comprising a first vaccine comprising a non-replicating immunogen of porcine circo virus type 2 (PCV2) and a non-replicating immunogen of Mycoplasma hyopneumoniae, and separately a second vaccine comprising a live attenuated PRRS virus; wherein an associated separate injection of the first vaccine and the second vaccine into a tissue of the pig at a first and a second injection site are at most 5 cm apart from each other.

12. A method of prophylactically treating a pig against an infection with porcine circovirus type 2 (PCV2), an infection with Mycoplasma hyopneumoniae and an infection with PRRS virus by injecting into a tissue of the pig at a first and a second injection site respectively, an associated separate injection of a first vaccine comprising a non-replicating immunogen of porcine circo virus type 2 (PCV2) and a non-replicating immunogen of Mycoplasma hyopneumoniae, and a second vaccine comprising a live attenuated porcine reproductive and respiratory syndrome (PRRS) virus; wherein the first and second injection sites are at most 5 cm apart from each other.

13. A vaccine comprising either: a non-replicating immunogen of porcine circo virus type 2 (PCV2) and a non-replicating immunogen of Mycoplasma hyopneumoniae, for use in prophylactically treating a pig against an infection with porcine circo virus type 2, an infection with Mycoplasma hyopneumoniae and an infection with PRRS virus, by associated separate injection of the vaccine with a second vaccine comprising a live attenuated porcine reproductive and respiratory syndrome (PRRS) virus into a tissue of the pig at a first and a second injection site respectively, or a live attenuated porcine reproductive and respiratory syndrome (PRRS) virus, for use in prophylactically treating an animal against an infection with porcine circo virus type 2, an infection with Mycoplasma hyopneumoniae and an infection with PRRS virus, by associated separate injection of the vaccine with a second vaccine comprising a non-replicating immunogen of porcine circo virus type 2 (PCV2) and a non-replicating immunogen of Mycoplasma hyopneumoniae into a tissue of the pig at a first and a second injection site respectively, wherein the first and second injection sites are at most 5 cm apart from each other.

14. (canceled)

Description

EXAMPLE

[0031] Objective

[0032] The objective of this experiment was to determine the efficacy of different mixed and non-mixed combinations of PCV-2, Mhyo and PRRS vaccines in piglets at three weeks of age, determined by PCV2 challenge two weeks post vaccination. PRRS and Mhyo efficacy were evaluated based on serology.

[0033] Study Design

[0034] Thirty pigs were used for this study. Three groups of 10 animals were vaccinated at the age of three weeks (+/− three days). Group 1 received a single dose of 2 ml of the commercial vaccine Porcilis PCV M Hyo (comprising baculo expressed ORF2 protein of PCV2 and an Mhyo bacterin) applied intramuscularly with a hypodermic syringe in the right side of neck, and a single dose of 2 ml the commercial vaccine Porcilis PRRS (comprising live attenuated PRRS virus, type I) applied intramuscularly with a hypodermic syringe at the same side of the neck, 3 cm apart from the other site of administration. Group 2 received the same vaccinations, but the PCV MHyo vaccine was administered at the right side of the neck, whereas the PRRS vaccine was administered at the left side of the neck. Group 3 was vaccinated with the PRRS vaccine only, serving as the PCV-2 challenge control. At two weeks post vaccination (5 weeks of age) all animals were challenged using a dose of 5.8 log.sub.10 TCID.sub.50 of wild-type PCV-2b challenge virus, applied intranasally.

[0035] Three weeks post challenge, all animals were necropsied and inguinal lymph node, mesenteric lymph node, tonsil and lung were sampled for the detection of PCV-2 nucleic acid. After vaccination, all piglets were observed daily for clinical signs. Serum samples were collected on the day of vaccination and 2, 4 and 5 weeks later (at the time of necropsy). Samples were tested for presence of PCV-2 viral nucleic acid and for antibodies against PRRS and M hyo.

[0036] Results

[0037] The vaccines were safe to administer, no unacceptable side effects were seen.

[0038] The PCV-2 results showed a consistent indication of an improved protection against the PCV-2 challenge. Results are indicated in the tables 1-3 below. As can be seen, the viraemia (at SD28 and 35), but in particular the viral load in various primary tissues and the IHC score seemed to be overall improved for Group 1, the group who received the two vaccines in close proximity, i.e. within 5 cm apart, in particular around 3 cm apart.

TABLE-US-00001 TABLE 1 PCV2 viraemia (qPCR) Titre (log10) SD0 SD13 SD28 SD35 1. PCV M HYO + PRRS (same side) 0.00 0.24 2.10 1.15 2. PCV M HYO + PRRS (diff sides) 0.00 0.20 2.38 1.40 3. Control (PRRS) 0.00 0.65 4.64 3.28

TABLE-US-00002 TABLE 2 PCV2 viral load in tissues (log10 copies/ul) Tonsil Lung IngLN MesLN 1. PCV M HYO + PRRS (same side) 4.58 4.42 3.97 3.86 2. PCV M HYO + PRRS (diff sides) 5.28 4.77 4.83 4.06 3. Control (PRRS) 7.41 6.32 6.71 6.89

TABLE-US-00003 TABLE 3 Average IHC Scores Tonsil IngLN MesLN Total 1. PCV M HYO + PRRS (same side) 0.3 0.3 0.4 1.0 2. PCV M HYO + PRRS (diff sides) 0.7 0.4 0.4 1.5 3. Control (PRRS) 2.5 2.3 2.4 7.2

[0039] Also, the Mhyo results appeared to be improved for Group 1. The results are indicated here below in tables 4 and 5. Seroconversion was sooner and the obtained S/P ratio was also consistently higher.

TABLE-US-00004 TABLE 4 Percentage of seroconverted animals SD0 SD13 SD28 SD35 1. PCV M HYO + PRRS (same side) 0 20 90 90 2. PCV M HYO + PRRS (diff sides) 0 0 40 80 3. Control (PRRS) 0 0 0 0

TABLE-US-00005 TABLE 5 Immune response to M Hyo S/P SD0 SD13 SD28 SD35 1. PCV M HYO + PRRS (same side) 0.04 0.19 0.86 1.05 2. PCV M HYO + PRRS (diff sides) 0.07 0.11 0.57 0.71 3. Control (PRRS) 0.15 0.08 0.03 0.01

[0040] Also, the PRRS results appeared to be improved for Group 1. The results are indicated here below in table 6. The obtained S/P ration was consistently higher for Group 1.

TABLE-US-00006 TABLE 6 Immune response to PRRS (ELISA) S/P SD0 SD13 SD28 SD35 1. PCV M HYO + PRRS (same side) 0.29 0.41 1.28 1.10 2. PCV M HYO + PRRS (diff sides) 0.35 0.30 0.80 0.89 3. Control (PRRS) 0.38 0.34 0.63 0.52

[0041] In short, the associated separate injection of the PCV Mhyo vaccine and the PRRS vaccine in close proximity led to an overall better immune response when compared to administering both vaccines at different sides of the neck. Given the fact that the effect was present for all types of antigen, either a subunit antigen (ORF2 protein of PCV2), a bacterin (Mhyo) or a live attenuated virus (PRRS), the exact choice of antigen appears to be non-critical. It is thus believed that at least when the first vaccine comprises immunogen of the same basic type as the exemplified, i.e. non-replicating immunogen typically inducing a humoral immune response, and the second vaccine comprises live attenuated PRRS virus, the advantages of the invention can be obtained.