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Chapter 4
Results
CHAPTER
4 RESULTS
Avian influenza (AI) is a serious disease of poultry, resulting in severe mortality in chickens and major disruption to production and trade (FAO, 2004). LPAI viruses cause respiratory and gastrointestinal infections without infecting the meat. By contrast, HPAI viruses produce infection of respiratory and gastrointestinal tracts and virus is present in the meat and internal contents of eggs during the acute stages of the infection (Swayne, 2004). Vaccines have been used as a tool in LPAI and HPAI eradication strategy. Avian influenza (AI) vaccines can prevent clinical signs and death in poultry, increase resistance to birds to infection, and decrease the amount of virus shed in the environment (Swayne, 2004) In the present study thirty commercial layers were divided into three groups, T1, T2 and T3 with 10 birds in each group. Group T1 served as control, Group T2 was immunized with conventional Avian influenza (AI) oil-based vaccines 0.5 ml/ bird and Group T3 was immunized with Avian influenza (AI) Liposomal vaccines 0.5 ml/ bird through sub/cut injection. Blood samples were taken and serum was separated at day 0, 7,14,21,28 and day 35. Each time at least 6 samples were taken for antibody titration through HI and AGPT. All the data for the experiment was analyzed by using the one way analysis of variance (ANOVA) (Table; 20 & 21). HI AND AGPT TITRES OF SERUM SAMPLES AT 0 DAY HI and AGPT titre were employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. Geometric mean titres (GMT) of birds in T1, T2 and T3 by HI and AGPT were 4+1.02 at day 0. The GMT of the entire groups were low and were considered as the background reading. No significant difference was observed in the titres at day 0 in all the groups (Table 2; Fig. 2). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
48
Chapter 4
Results
Table 2. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 0 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES 1:8 1:4 1:2
1:8 1:4 1:2
AGPT TITRES 1:8 1:4 1:2
1:8 1:4 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A
EM 2.00 + 0.89 GMT 4 +1.021 A
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) While values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
49
Chapter 4
Results
HI and AGPT titres at day 0 HI titres 4.5
4
4
AGPT titres
4
4
4
4
4 3.5 3 T i tr e s
2.5 2 1.5 1 0.5 0
control
oil based
liposomal
HI titres
4
4
4
AGPT titres
4
4
4
Groups
Figure 2. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on Day 0
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
50
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 7 DAY
Haemagglutination Inhibition (HI) titres were evaluated to compare immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Avian Influenza (AI) Liposomal vaccine (0.5 ml) subcutaneously. Looking in to the data the GMT titres were marginal in group T1 (7 + 0.86) whereas, it was higher in T2 (32 + 0.722).The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly higher in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was quite significant (64 + 0.722) as compared with T1 (7 + 0.86) and T2 (32 + 0.722).The data comparatively indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 3; Fig. 3). AGPT TITRES OF SERUM SAMPLES AT 7 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Avian Influenza (AI) Liposomal vaccine (0.5 ml) subcutaneously. The GMT titre at day 14 in T1, T2 and T3 were observed by AGPT. GMT titre were low in group T1 (4 + 1.021), whereas it was high in T2 (32 + 0.722). The effect of Avian Influenza (AI) Oil based vaccine in T2 was significantly higher in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (64 + 0.722) as compared with T1 (4 + 1.021) and T2 (32 + 0.722). The data comparatively indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 3; Fig. 3). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7 51
Chapter 4
Results
Table 3. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 7 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES 1:8 1:4 1:16
1:8 1:4 1:8
AGPT TITRES 1:4 1:8 1:2
1:2 1:4 1:8
EM 2.83 + 0752 GMT 7 + 0.859 A 1:16 1:32 1:32 1:32 1:32 1:64
EM 2.00 + 0.894 GMT 4 + 1.021 A 1:16 1:32 1:32 1:32 1:32 1:64
EM 5.00 + 0.632 GMT 32 + 0.722 B 1:128 1:64 1:32 1:64 1:64 1:64
EM 5.00 + 0.632 GMT 32 + 0.722 B 1:128 1:64 1:32 1:64 1:64 1:64
EM 6.00 + 0.632 GMT 64 + 0.722 C
EM 6.00 + 0.632 GMT 64 + 0.722 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
52
Chapter 4
Results
HI and AGPT titres at Day 7 70
64
HI titres
60
64
AGPT titres
50 T i tr e s
40 32
32
30 20 10 0
7
4
control
oil based
liposomal
HI titres
7
32
64
AGPT titres
4
32
64
Groups
Figure 3. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 7.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
53
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 14 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. In comparison the GMT titres were marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (128 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (256 + 0.722) as compared with T1 (7 + 0.859) and T2 (128 + 0.722). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 4; Fig. 4). AGPT TITRES OF SERUM SAMPLES AT 14 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. Immune response of different treatments through GMT titre at day 14 in T1, T2 and T3 were observed by AGPT. GMT titres were marginal in group T1 (6.79 + 1.021) whereas, it was quite higher in T2 (128 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (256 + 0.722) as compared with T1 (6.79 + 1.021) and T2 (128 + 0.722). The data fairly indicated that immune response in term of GMT was well established with AI Liposomal vaccine as compared to AI Oil based vaccine (Table 4; Fig. 4). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
54
Chapter 4
Results
Table 4. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 14 post treatment (n = 6)
TREATMENT
T1 (CONTROL) GMT + SDM
T2 (AI OIL BASED) GMT + SDM
T3 (AI LIPOSOMAL) GMT + SDM
HI TITRES 1:8 1:4 1:16
1:8 1:4 1:8
AGPT TITRES 1:4 1:8 1:16
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:128 1:128 1:64 1:128 1:256 1:128
EM 3.00 + 0.894 GMT 6.79 + 0.859 A 1:128 1:128 1:64 1:128 1:256 1:128
EM 7.00 + 0.632 GMT 128 + 0.722 B 1:512 1:256 1:128 1:256 1:256 1:256
EM 7.00 + 0.632 GMT 128 + 0.722 B 1:512 1:256 1:128 1:256 1:256 1:256
EM 8.00 + 0.632 GMT 256 + 0.722 C
EM 8.00 + 0.632 GMT 256 + 0.722 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
55
Chapter 4
Results
HI and AGPT titres at Day 14 300 HI titre (GMT) AGPT titre (GMT)
256
256
250
Titre
200
150 128
128
100
50
8
7 0
control
oil based
liposomal
HI titre (GMT)
7
128
256
AGPT titre (GMT)
8
128
256
Groups
Figure 4. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 14.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
56
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 21 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with AI Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titres by HI on day 21 were marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (128 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high as compared to control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant with GMT (512 + 0.722) as compared with T1 (7 + .86) and T2 (256 + 0.722). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table5; Fig. 5). AGPT TITRES OF SERUM SAMPLES AT 21 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. GMT titre in T1, T2 and T3 were observed at day 21 by AGPT. The GMT titres were marginal in group T1 (8 + 1.021) whereas, it was quite higher in T2 (256 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (512 + 0.722) as compared to T1 (7 + 0.859) and T2 (128 + 0.722). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Oil based vaccine (Table 5; Fig. 5). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
57
Chapter 4
Results
Table 5. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 21 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES 1:8 1:4 1:16
1:8 1:4 1:8
AGPT TITRES 1:4 1:8 1:16
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:128 1:128 1:64 1:128 1:256 1:128
EM 3.00 + 0.894 GMT 8.00 + 1.021 A 1:512 1:256 1:128 1:256 1:256 1:256
EM 7.00 + 0.632 GMT 128 + 0.722 B 1:512 1:512 1:256 1:512 1:1024 1:512
EM 8.00 + 0.632 GMT 256 + 0.722 B 1:512 1:512 1:256 1:512 1:1024 1:512
EM 9.00 + 0.632 GMT 512 + 0.722 C
EM 9.00 + 0.632 GMT 512 + 0.722 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
58
Chapter 4
Results
HI and AGPT titres at Day 21 600
HI titres
512
512
AGPT titres
500
T itr e s
400 300
256
200 128 100 7 0
8
control
oil based
liposomal
HI titres
7
128
512
AGPT titres
8
256
512
Groups
Figure 5. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 21.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
59
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 28 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titres by HI at day 28 were marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (445.7 + 0.46). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (891.4 + 0.46) as compared with T1 (7 + 0.86) and T2 (445.7 + 0.46). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 6; Fig. 6). AGPT TITRES OF SERUM SAMPLES AT 28 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titres in T1, T2 and T3 at day 28 were observed by AGPT. GMT titres were marginal in group T1 (8 + 1.022), whereas it was quite higher in T2 (445.7 + 0.46). The effect of AI oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (776 + 0.056) as compared with T1 (8 + 1.022) and T2 (445.7 + 0.46). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Oil based vaccine (Table 6; Fig. 6). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
60
Chapter 4
Results
Table 6. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 28 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES
AGPT TITRES
1:8 1:16 1:4
1:4 1:8 1:16
1:8 1:8 1:4
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:512 1:512 1:256 1:512 1:512 1:512
EM 3.00 + 0.894 GMT 8 + 1.022 A 1:512 1:512 1:256 1:512 1:512 1:512
EM 8.83 + 0.408 GMT 445.7 + 0.466 B 1:1024 1:1024 1:1024 1:1024 1:1024 1:512
EM 8.83 + 0.408 GMT 445.7 + 0.466 B 1:1024 1:1024 1:512 1:1024 1:1024 1:512
EM 9.83 + 0.408 GMT 891.4 + 0.466 C
EM 9.60 + 0.516 GMT 776 + 0.589 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) While values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
61
Chapter 4
Results
HI and AGPT titres at Day 28 1000
HI titres
900
891.4
AGPT titres
800
776
700 T itr e s
600 445.7 445.7
500 400 300 200 100 0
7
8
control
oil based
liposomal
HI titres
7
445.7
891.4
AGPT titres
8
445.7
776
Groups
Figure 6. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 28.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
62
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 35 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. Temporal response relationship were noted, GMT of 7 + 0.86 (T1), 548.7 + 0.46 (T2) and 1097.5 + 0.46 (T3) by HI were recorded at day 35. GMT titre was marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (548.7 + 0.46). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (1097.5 + 0.46) as compared with T1 (7 + 0.86) and T2 (548.7 + 0.46). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 7; Fig. 7). AGPT TITRES OF SERUM SAMPLES AT 35 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titre in T1, T2 and T3 were observed by AGPT at day 35. GMT titres were marginal in group T1 (8 + 1.022), whereas it was quite higher in T2 (630.3 + 0.56). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (891.4 + 0.46) as compared with T1 (8 + 1.022) and T2 (630.3 + 0.56). The data fairly indicated that immune response in term of GMT was well established with AI Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 7; Fig. 7). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
63
Chapter 4
Results
Table 7. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 35 post treatment (n = 6)
TREATMENT
T1 (CONTROL) GMT + SDM
T2 (AI OIL BASED) GMT + SDM
T3 (AI LIPOSOMAL) GMT + SDM
HI TITRES 1:8 1:16 1:4
1:8 1:8 1:4
AGPT TITRES 1:4 1:8 1:16
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:512 1:512 1:1024 1:512 1:512 1:512
EM 3.00 + 0.894 GMT 8+ 1.021 A 1:512 1:512 1:1024 1:512 1:512 1:1024
EM 9.16 + 0.408 GMT 548.7 + 0.466 B 1:1024 1:1024 1:1024 1:1024 1:1024 1:2048
EM 9.33 + 0.516 GMT 630.3 + 0.589 B 1:1024 1:1024 1:1024 1:1024 1:1024 1:512
EM 10.16 + 0.408 GMT 1097.5 +0.466 C
EM 9.83 + 0.408 GMT 891.4 + 0.466 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
64
Chapter 4
Results
HI and AGPT titres at Day 35 1200
1097.5
HI titres AGPT titres
1000
891.4
T i tr e s
800 630.3 548.7
600 400 200 8
7 0
control
oil based
liposomal
HI titres
7
548.7
1097.5
AGPT titres
8
630.3
891.4
Groups
Figure 7. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 35.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
65
Results
CHAPTER
4 RESULTS
Avian influenza (AI) is a serious disease of poultry, resulting in severe mortality in chickens and major disruption to production and trade (FAO, 2004). LPAI viruses cause respiratory and gastrointestinal infections without infecting the meat. By contrast, HPAI viruses produce infection of respiratory and gastrointestinal tracts and virus is present in the meat and internal contents of eggs during the acute stages of the infection (Swayne, 2004). Vaccines have been used as a tool in LPAI and HPAI eradication strategy. Avian influenza (AI) vaccines can prevent clinical signs and death in poultry, increase resistance to birds to infection, and decrease the amount of virus shed in the environment (Swayne, 2004) In the present study thirty commercial layers were divided into three groups, T1, T2 and T3 with 10 birds in each group. Group T1 served as control, Group T2 was immunized with conventional Avian influenza (AI) oil-based vaccines 0.5 ml/ bird and Group T3 was immunized with Avian influenza (AI) Liposomal vaccines 0.5 ml/ bird through sub/cut injection. Blood samples were taken and serum was separated at day 0, 7,14,21,28 and day 35. Each time at least 6 samples were taken for antibody titration through HI and AGPT. All the data for the experiment was analyzed by using the one way analysis of variance (ANOVA) (Table; 20 & 21). HI AND AGPT TITRES OF SERUM SAMPLES AT 0 DAY HI and AGPT titre were employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. Geometric mean titres (GMT) of birds in T1, T2 and T3 by HI and AGPT were 4+1.02 at day 0. The GMT of the entire groups were low and were considered as the background reading. No significant difference was observed in the titres at day 0 in all the groups (Table 2; Fig. 2). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
48
Chapter 4
Results
Table 2. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 0 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES 1:8 1:4 1:2
1:8 1:4 1:2
AGPT TITRES 1:8 1:4 1:2
1:8 1:4 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A 1:8 1:8 1:4 1:4 1:2 1:2
EM 2.00 + 0.89 GMT 4 +1.021 A
EM 2.00 + 0.89 GMT 4 +1.021 A
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) While values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
49
Chapter 4
Results
HI and AGPT titres at day 0 HI titres 4.5
4
4
AGPT titres
4
4
4
4
4 3.5 3 T i tr e s
2.5 2 1.5 1 0.5 0
control
oil based
liposomal
HI titres
4
4
4
AGPT titres
4
4
4
Groups
Figure 2. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on Day 0
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
50
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 7 DAY
Haemagglutination Inhibition (HI) titres were evaluated to compare immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Avian Influenza (AI) Liposomal vaccine (0.5 ml) subcutaneously. Looking in to the data the GMT titres were marginal in group T1 (7 + 0.86) whereas, it was higher in T2 (32 + 0.722).The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly higher in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was quite significant (64 + 0.722) as compared with T1 (7 + 0.86) and T2 (32 + 0.722).The data comparatively indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 3; Fig. 3). AGPT TITRES OF SERUM SAMPLES AT 7 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Avian Influenza (AI) Liposomal vaccine (0.5 ml) subcutaneously. The GMT titre at day 14 in T1, T2 and T3 were observed by AGPT. GMT titre were low in group T1 (4 + 1.021), whereas it was high in T2 (32 + 0.722). The effect of Avian Influenza (AI) Oil based vaccine in T2 was significantly higher in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (64 + 0.722) as compared with T1 (4 + 1.021) and T2 (32 + 0.722). The data comparatively indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 3; Fig. 3). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7 51
Chapter 4
Results
Table 3. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 7 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES 1:8 1:4 1:16
1:8 1:4 1:8
AGPT TITRES 1:4 1:8 1:2
1:2 1:4 1:8
EM 2.83 + 0752 GMT 7 + 0.859 A 1:16 1:32 1:32 1:32 1:32 1:64
EM 2.00 + 0.894 GMT 4 + 1.021 A 1:16 1:32 1:32 1:32 1:32 1:64
EM 5.00 + 0.632 GMT 32 + 0.722 B 1:128 1:64 1:32 1:64 1:64 1:64
EM 5.00 + 0.632 GMT 32 + 0.722 B 1:128 1:64 1:32 1:64 1:64 1:64
EM 6.00 + 0.632 GMT 64 + 0.722 C
EM 6.00 + 0.632 GMT 64 + 0.722 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
52
Chapter 4
Results
HI and AGPT titres at Day 7 70
64
HI titres
60
64
AGPT titres
50 T i tr e s
40 32
32
30 20 10 0
7
4
control
oil based
liposomal
HI titres
7
32
64
AGPT titres
4
32
64
Groups
Figure 3. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 7.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
53
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 14 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. In comparison the GMT titres were marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (128 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (256 + 0.722) as compared with T1 (7 + 0.859) and T2 (128 + 0.722). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 4; Fig. 4). AGPT TITRES OF SERUM SAMPLES AT 14 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. Immune response of different treatments through GMT titre at day 14 in T1, T2 and T3 were observed by AGPT. GMT titres were marginal in group T1 (6.79 + 1.021) whereas, it was quite higher in T2 (128 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (256 + 0.722) as compared with T1 (6.79 + 1.021) and T2 (128 + 0.722). The data fairly indicated that immune response in term of GMT was well established with AI Liposomal vaccine as compared to AI Oil based vaccine (Table 4; Fig. 4). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
54
Chapter 4
Results
Table 4. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 14 post treatment (n = 6)
TREATMENT
T1 (CONTROL) GMT + SDM
T2 (AI OIL BASED) GMT + SDM
T3 (AI LIPOSOMAL) GMT + SDM
HI TITRES 1:8 1:4 1:16
1:8 1:4 1:8
AGPT TITRES 1:4 1:8 1:16
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:128 1:128 1:64 1:128 1:256 1:128
EM 3.00 + 0.894 GMT 6.79 + 0.859 A 1:128 1:128 1:64 1:128 1:256 1:128
EM 7.00 + 0.632 GMT 128 + 0.722 B 1:512 1:256 1:128 1:256 1:256 1:256
EM 7.00 + 0.632 GMT 128 + 0.722 B 1:512 1:256 1:128 1:256 1:256 1:256
EM 8.00 + 0.632 GMT 256 + 0.722 C
EM 8.00 + 0.632 GMT 256 + 0.722 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
55
Chapter 4
Results
HI and AGPT titres at Day 14 300 HI titre (GMT) AGPT titre (GMT)
256
256
250
Titre
200
150 128
128
100
50
8
7 0
control
oil based
liposomal
HI titre (GMT)
7
128
256
AGPT titre (GMT)
8
128
256
Groups
Figure 4. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 14.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
56
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 21 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with AI Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titres by HI on day 21 were marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (128 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high as compared to control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant with GMT (512 + 0.722) as compared with T1 (7 + .86) and T2 (256 + 0.722). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table5; Fig. 5). AGPT TITRES OF SERUM SAMPLES AT 21 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. GMT titre in T1, T2 and T3 were observed at day 21 by AGPT. The GMT titres were marginal in group T1 (8 + 1.021) whereas, it was quite higher in T2 (256 + 0.722). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (512 + 0.722) as compared to T1 (7 + 0.859) and T2 (128 + 0.722). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Oil based vaccine (Table 5; Fig. 5). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
57
Chapter 4
Results
Table 5. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 21 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES 1:8 1:4 1:16
1:8 1:4 1:8
AGPT TITRES 1:4 1:8 1:16
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:128 1:128 1:64 1:128 1:256 1:128
EM 3.00 + 0.894 GMT 8.00 + 1.021 A 1:512 1:256 1:128 1:256 1:256 1:256
EM 7.00 + 0.632 GMT 128 + 0.722 B 1:512 1:512 1:256 1:512 1:1024 1:512
EM 8.00 + 0.632 GMT 256 + 0.722 B 1:512 1:512 1:256 1:512 1:1024 1:512
EM 9.00 + 0.632 GMT 512 + 0.722 C
EM 9.00 + 0.632 GMT 512 + 0.722 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
58
Chapter 4
Results
HI and AGPT titres at Day 21 600
HI titres
512
512
AGPT titres
500
T itr e s
400 300
256
200 128 100 7 0
8
control
oil based
liposomal
HI titres
7
128
512
AGPT titres
8
256
512
Groups
Figure 5. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 21.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
59
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 28 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titres by HI at day 28 were marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (445.7 + 0.46). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (891.4 + 0.46) as compared with T1 (7 + 0.86) and T2 (445.7 + 0.46). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 6; Fig. 6). AGPT TITRES OF SERUM SAMPLES AT 28 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titres in T1, T2 and T3 at day 28 were observed by AGPT. GMT titres were marginal in group T1 (8 + 1.022), whereas it was quite higher in T2 (445.7 + 0.46). The effect of AI oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (776 + 0.056) as compared with T1 (8 + 1.022) and T2 (445.7 + 0.46). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Oil based vaccine (Table 6; Fig. 6). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
60
Chapter 4
Results
Table 6. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 28 post treatment (n = 6)
TREATMENT
T1 (CONTROL)
GMT + SDM
T2 (AI OIL BASED)
GMT + SDM
T3 (AI LIPOSOMAL)
GMT + SDM
HI TITRES
AGPT TITRES
1:8 1:16 1:4
1:4 1:8 1:16
1:8 1:8 1:4
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:512 1:512 1:256 1:512 1:512 1:512
EM 3.00 + 0.894 GMT 8 + 1.022 A 1:512 1:512 1:256 1:512 1:512 1:512
EM 8.83 + 0.408 GMT 445.7 + 0.466 B 1:1024 1:1024 1:1024 1:1024 1:1024 1:512
EM 8.83 + 0.408 GMT 445.7 + 0.466 B 1:1024 1:1024 1:512 1:1024 1:1024 1:512
EM 9.83 + 0.408 GMT 891.4 + 0.466 C
EM 9.60 + 0.516 GMT 776 + 0.589 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) While values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
61
Chapter 4
Results
HI and AGPT titres at Day 28 1000
HI titres
900
891.4
AGPT titres
800
776
700 T itr e s
600 445.7 445.7
500 400 300 200 100 0
7
8
control
oil based
liposomal
HI titres
7
445.7
891.4
AGPT titres
8
445.7
776
Groups
Figure 6. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 28.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
62
Chapter 4
Results
HI TITRES OF SERUM SAMPLES AT 35 DAY
Haemagglutination Inhibition (HI) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. Temporal response relationship were noted, GMT of 7 + 0.86 (T1), 548.7 + 0.46 (T2) and 1097.5 + 0.46 (T3) by HI were recorded at day 35. GMT titre was marginal in group T1 (7 + 0.86) whereas, it was quite higher in T2 (548.7 + 0.46). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (1097.5 + 0.46) as compared with T1 (7 + 0.86) and T2 (548.7 + 0.46). The data fairly indicated that immune response in term of GMT was well established with Avian Influenza (AI) Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 7; Fig. 7). AGPT TITRES OF SERUM SAMPLES AT 35 DAY
Agar gel precipitation test (AGPT) titre was employed to evaluate the comparative immunological response of commercial Oil based and Liposomal vaccines of Avian Influenza (AI) in commercial Layers. In treatment group T1, birds were served as control group; T2, birds were vaccinated with Avian Influenza (AI) Oil based vaccine (0.5 ml) subcutaneously and T3, birds were vaccinated with Liposomal vaccine (0.5 ml) subcutaneously. The GMT titre in T1, T2 and T3 were observed by AGPT at day 35. GMT titres were marginal in group T1 (8 + 1.022), whereas it was quite higher in T2 (630.3 + 0.56). The effect of Avian Influenza (AI) oil based vaccine in T2 was significantly high in comparison with control group (T1). However Avian Influenza (AI) Liposomal vaccinated group (T3) was highly significant (891.4 + 0.46) as compared with T1 (8 + 1.022) and T2 (630.3 + 0.56). The data fairly indicated that immune response in term of GMT was well established with AI Liposomal vaccine as compared to Avian Influenza (AI) Oil based vaccine (Table 7; Fig. 7). Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
63
Chapter 4
Results
Table 7. HI and AGPT titres of Avian Influenza antibodies in birds, with different regimens of immunization at day 35 post treatment (n = 6)
TREATMENT
T1 (CONTROL) GMT + SDM
T2 (AI OIL BASED) GMT + SDM
T3 (AI LIPOSOMAL) GMT + SDM
HI TITRES 1:8 1:16 1:4
1:8 1:8 1:4
AGPT TITRES 1:4 1:8 1:16
1:16 1:4 1:8
EM 2.83 + 0.752 GMT 7 + 0.859 A 1:512 1:512 1:1024 1:512 1:512 1:512
EM 3.00 + 0.894 GMT 8+ 1.021 A 1:512 1:512 1:1024 1:512 1:512 1:1024
EM 9.16 + 0.408 GMT 548.7 + 0.466 B 1:1024 1:1024 1:1024 1:1024 1:1024 1:2048
EM 9.33 + 0.516 GMT 630.3 + 0.589 B 1:1024 1:1024 1:1024 1:1024 1:1024 1:512
EM 10.16 + 0.408 GMT 1097.5 +0.466 C
EM 9.83 + 0.408 GMT 891.4 + 0.466 C
T1: Birds served as control T2: Birds vaccinated with Avian Influenza Oil Based Vaccine 0.5 ml subcutaneously T3: Birds vaccinated with Avian Influenza Liposomal Vaccine 0.5 ml subcutaneously A-C: Values with similar superscript do not differ statistically (P<0.05) while values with different superscript differ significantly
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
64
Chapter 4
Results
HI and AGPT titres at Day 35 1200
1097.5
HI titres AGPT titres
1000
891.4
T i tr e s
800 630.3 548.7
600 400 200 8
7 0
control
oil based
liposomal
HI titres
7
548.7
1097.5
AGPT titres
8
630.3
891.4
Groups
Figure 7. Graph showing the HI and AGPT titre of control, Oil based and Liposomal vaccinated Groups on day 35.
Comparative Immunological Studies on Commercial Oil Based and Liposomal Vaccines of Avian Influenza H7
65
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