Bivalent Coryza Vaccine KITASATO

Kume Katsumi The Kitasato Institute Tokyo, Japan

Bivalent Coryza Vaccine KITASATO In 1981, Kitasato Institute developed a bivalent coryza vaccine KITASATO (strains 221 and H-18). Since then, this vaccine has been available not only in Japan but also in over 30 foreign countries having coryza problems.

Causative Agent of Infectious Coryza

Only encapsulated organisms of H. paragallinarum having antigenicity and immunogenicity can cause infectious coryza. Iridescence of colony strictly reflects presence of capsule on the surface of organisms. Therefore, selection of encapsulated organisms from high iridescent type of colony is very important. Never use blood agar plate for isolation purpose.

Encapsulated organism of Haemophilus paragallinarum

Iridescent type of colony

Bivalent Coryza Vaccine KITASATO

Encapsulated organisms involved in the Bivalent Coryza Vaccine KITASATO

Quality of Infectious Coryza Vaccine (ICV) is easily distinguished. Organisms involved in ICV must be uniformly (short rods). If ICVs involves various shapes of organisms (long rods and filaments), such ICVs are not only ineffective but also harmful to chickens.

Duration of HI Antibodies After Injection of Coryza Vaccine

80 40 HI titer 20 10 5

*
5 6 8 10 12 15 Weeks of age 20 25

Immunized at 5 weeks of age with 0.5ml/chicken of vaccine

* Minimum protection level

Duration of HI Antibodies After Injection of Infectious Coryza Vaccine

HI antibodies arise 7 to 10 days after injection of ICV, reached peak value at 3 to 4 weeks after vaccination, and then decrease gradually. In case of one shot, immunity persist for about 5 months after vaccination.

Duration of HI Antibodies After Injection of Coryza Vaccine

2nd shot 2nd shot 2nd shot
1st shot Immunization program Group I (5w, 8w) Group II (5w, 15w) Group III (5w, 20w)

160 80 Hi titer 40 20 10 5 5

*
8 10 15 Weeks of age

20

25

30

* Minimum protection level

Duration of HI Antibodies After Injection of Coryza Vaccine

Interval between first and second shot is very important for duration of immunity. Interval must be kept between 9 to 11 weeks, when use bivalent coryza vaccine KITASATO.

Serotyping of H.paragallinarum by Serotype-Specific HA-L Hemagglutinin

Serotype Classified Reference Origin of by serotype-specific strain the strain hamagglutinin Factor of HA-L hemagglutinin Specific Common

HA-1 (A) HA-2 (A)* HA-3 (A)

221 Japan 2403 Germany E-3C Brazil

I1 I2 I3 II 1 II 2 II 3 III 1

I 4, I - 5 I 4, I - 5 I4 II 4, II - 5 II 4, II - 5 II 4

HA-4 (C) H-18 Japan HA-5 (C) Modesto U.S.A HA-6 (C) SA-3 South Africa HA-7 (B)
* : Page (B)

2671 Germany

At least 7 serotype strains are classified by serotype-specific HA-L hemagglutinin system.

Hemagglutination-Inhibition (HI) AntibodyProducing Antigenicity of H.paragallinarum

HI antibody titer against HA-L antigen Group Antiserum (hi-NT) Serotype 221 2403 E-3C H-18 Modesto SA-3 2671

221
I

1(A) (B) ND 2(C) ND (B)

2,560 2,560 320 <10 <10 <10 <10

2,560 2,560 320 <10 <10 <10 <10

320 640 1,280 <10 <10 <10 <10

<10 <10 <10 1,280 1,280 20 <10

<10 <10 <10 1,280 2,560 320 <10

<10 <10 <10 80 160 2,560 <10

<10 <10 <10 <10 <10 <10 2,560

2403 E-3C H-18

II

Modesto 2(C) SA-3

III

2671

Seven serotype strains are classified into 3 groups according to HI antibody-producing antigenicity.

Cross-Protection Studies Among the Seven Serotype Strains of H.paragallinarum

Cross-protection studies indicate the presence of 3 immunogroups among the 7 serotype strains. However, bivalent coryza vaccine KITASATO composed of serotype HA-1 (A) and HA-4 (C) strains can protect against any of the 7 serotype strains. Therefore, it is not necessary to use trivalent vaccines, if you can apply high quality bivalent vaccines, even in the presence of serotype B strains in the field.

Cross-Protection Studies Among the Seven Serotype Strains of H.paragallinarum

Immunogen Serotype 221 Challenge strains and protective activity *1 2403 E-3C H-18 Modesto SA-3 2671

221 2403 E-3C H-18 SA-3 2671 221 + H-18

HA-1 HA-2 HA-3 HA-4 HA-6 HA-7 HA-1 + HA-4

++ ++ ++ ++ ++

+ ++ + + + + ++

+ + ++ + ++

++ + + + ++

+ ++ + ++

+ + ++ +

+ + + + ++ +

Modesto HA-5

*1) ++ = Protection rate above 70% (injected once) + = Protection rate above 70% (injected twice)

- = No protection (infected twice)

Distribution of H.paragallinarum Serotype Strain in the World Serotype HA-1 and HA-4 strains are predominant type world-wide.

Distribution of H.paragallinarum Serotype Strain in the World (Tested by the Kitasato Institute) Serotype defined by hemagglutination (Serotype defined by agglutination) HA-1 ( A ) HA-4 ( C ) HA-1 ( A ) HA-4 ( C ) HA-1 ( A ) HA-3 ( A ) HA-4 ( C ) HA-7 ( B ) HA-1 ( A ) HA-4 ( C ) HA-1 ( A ) HA-4 ( C ) HA-1 ( A ) HA-4 ( C ) HA-1 ( A ) HA-4 ( C ) HA-1 ( A ) HA-4 ( C ) HA-7 (B) HA-1 ( A ) HA-4 ( C ) HA-6 ( C ) HA-1 ( A ) HA-4 ( C ) HA-7 ( B ) HA-1 ( A ) HA-4 ( C ) HA-7 ( B )

Country Japan Taiwan Brazil Philippines Malaysia Indonesia Venezuela Thailand Mexico South Africa Argentina Chaina

The Correlation Between the Protective Capability and HI Antibody Titers of the Coryza Vaccine
HI titer At challenge No.of chickens Protection rate (%) No.of chickens Protection rate (%) H-18 (Serotype HA-4) vaccine 221 (Serotype HA-1) vaccine

Immunized

<1:5 1:5 1:10 1:20 1:40 1:80 1:160 > 1:320 = <1:5

17 19 31 27 24 9 12 11 20

47 94.7 96.8 100 100 100 100 100 0

28 22 29 29 22 32 34 35 15

50 91 100 100 100 100 100 100 0

Control

The Correlation Between the Protective Capability and HI Antibody Titers of the Coryza Vaccine Chickens having HI antibodies above 5 are protected against challenge exposure. Minimum protection level of ICV is 1:5. Efficacy of ICV is easily measured by the HI test.

Recovery of Challenge Organisms From the Nasal Cavities of Chickens Injected with Coryza Vaccine KITASATO

Chickens lacking HI antibodies (less than 1:5) at challenge suffer typical clinical signs of coryza, and a high number of organisms are recovered from these diseased chickens. Chickens having HI antibodies above 5 at challenge never show clinical signs during the observation period, and challenged organisms are easily cleared from these chickens without (above 1:20) or after slight (1:5 and 1:10) proliferation of challenge organisms. Presence of HI antibodies and the level of HI titer in immunized chickens are important factors in IC prevention.

10

Recovery of Challenge Organisms From the Nasal Cavities of Chickens Injected with Coryza Vaccine KITASATO 6
HI titers at 1 day before challenge

105 104

Viable cells b) /sample

103

<5 a) * <5 40 80 160 * 5 10 20 *

* *

102 101 >320 * 1 3 6 12 Hours Post-challenge 24 48

a) Non injected control chickens. b) Data are shown as an average of the organism numbers recovered from the chickens positive for the isolation.

Duration of Immunity in Chickens Injected with Coryza Vaccine KITASATO

Age injected (Dosage) 5 weeks (0.5ml) 5 weeks (1.0ml) 5 weeks (0.5ml) + 15 weeks (0.5ml) 5 weeks (0.5ml) + 15 weeks (1.0ml) Protective rate (%) after final injection 1 3 5 7 10 months 90 80 60 50 40 90 90 70 50 40 90 90 90 80 70

90

90

90

80

80

Coryza vaccine KITASATO can be applied 0.5ml (first) and 0.5ml (second) doses. However, we recommend a total of 1.5ml for breeding and highly contaminated farms.

Duration of HI Antibodies After Vaccination with Bivalent Coryza Vaccine KITASATO

80 First 40 HI antibody titers (GMT) 2nd 0.5ml+0.5ml 0.5ml+1.0ml

20

10 5

15

20 Weeks of age

35

50

Duration of Immunity After Injection with Bivalent Coryza Vaccine KITASATO Immunity evaluated by challenge test persists about 5 months (one shot) and nearly one year (two shots). Duration of HI antibodies in immunized chickens strongly supports our data.

Quality of Infectious Coryza Vaccines (ICVs)

High quality ICVs must produce HI antibodies against any of the serotype strains involved in Infectious Coryza vaccine. Both HI positive rate and geometric mean (GM) of HI titers in immunized group are very important for evaluation of ICVs quality.

Protection Rate (HI Antibody-Positive Rate) of Chickens 3 (Alumi gel) or 4 (oil) Weeks After Vaccination with Various Kinds of IC Vaccines
Serotype A

100 80 60 40

100 100 90

100

Serotype B Serotype C

60 50 30 30 20 0
A C 85.7 113.1 KITASATO A B C 3.5 <2.5 4.4 A A C 13.5 4.4 B A 3.5 C C 4.4 A B C 52.8 5.0 3.8 D

40 30

20 0
Serotype GMT of HI Maker

Production of HI antibodies against type A and type C after injection of Coryza Vaccine is very important. For long duration of immunity, HI antibodies must arise to a certain high level within 3 to 4 weeks after the first injection of Coryza Vaccine. To analyze this point, GMT of HI antibody is very important.

HI antibody-positive rate is also very important for selection of real bivalent or trivalent type of ICV. Kitasato Coryza Vaccine is only the real meaning of bivalent ICV among the test samples. Real bivalent ICV is effective against any of the known serotype of H.paragallinarum. So, it is not necessary to use trivalent ICVs.

Difference in Protective Potency of Coryza Vaccine After Different Injection Sites in Chickens

Site Leg muscle Breast muscle Neck subcutaneous

No. of chickens Protection rate (%) 40 40 40 87.5 52.5 32.5

Bivalent coryza vaccine KITASATO must be injected into leg muscle to get good immunity

Application Method of Bivalent Coryza Vaccine KITASATO

Recommended Vaccination Program (2 Patterns)

0.5ml 0.5ml 0.5~1ml 0.5~1ml

5~6 weeks of age

15~16 weeks of age

A 10-week-interval !! Inject exactly into leg muscle ! At 1st : The immune system is immature before 5 weeks of age. At 2nd : Vaccination should be done before starting egg production.