Proceedings 7th Chula. Univ. Vet. Sci. Ann. Con., 1 May, 2008.

P13 Detection of Active Immunity Level Against a Fowl Cholera Vaccine in

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Khaki Campbell Ducks (Anas platyrhynchos) by Microplate Agglutination Test

S. Khanda1 P. Khanda1 J. Sasipreeyajan1 N. Chansiripornchai1
1

Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand

Keywords: Duck, microplate agglutination test, fowl cholera Introduction

Fowl cholera (FC) is a disease caused by Pasteurella multocida. FC is frequently found especially during the seasonal changes from rainy to winter. FC can be prevented and controlled by using many measures: antibiotics, vaccines, good management and biosecurity (1). An inactivated FC vaccine has been produced by DLD, Thailand originating from strain 8:A. The vaccine is applied by intramuscular route for 1 ml/dose. The objective was to study the protective levels of the antibody by microtiter agglutination (MA) test. incubated for 37oC for 1 hr and kept at 4oC. The results were read after 24 hr, at 50% agglutination

Results and Discussion

Group 1 revealed the highest MA titers at 2 weeks after vaccination. Two weeks after challenge at 8 weeks old, the titers increase a and the highest titer levels were found at 4 weeks after challenge. Eighty percent of ducks in group 1 were alive after challenge. Group 2 showed an MA titer similar to those of ducks in group 1. The MA titers were peak at 4 weeks after 2nd vaccination. Eighty seven point five percent of ducks in group 2 could be alive after challenge. In the ducks in group 3, a positive control group, the MA titer was equal to 31.04 at 8 weeks old. All the ducks died after challenge (Table 1). In conclusion, MA tests can be used as a screening test for antibody detection against FC. This method can be applied in microplates instead of test tubes. This technique can save both antigen and serum. Anyhow, it needs to be further developed.

Materials and Method

Forty six Khaki Campbell ducks were raised from 1 day old. Group 1: ducks were vaccinated once at 4 weeks old and challenged at 8 weeks old. Group 2: ducks were vaccinated twice at 4 and 8 weeks old and challenged at 12 weeks old. Group 3: a positive control group, ducks were challenged at 8 weeks old. Blood was collected every week after 3 weeks old. Then, all the ducks were euthanized. Sera were kept at -20oC until use. P. multocida was prepared and concentrated at 3-5x103 cfu/ml and injected into the right leg of the ducks at 1 ml. Antigen preparation was performed by culturing P. multocida overnight, 37oC and adjusting to 109 cfu/ml, centrifuged at 5,000 rpm, 30 min. and then washed. The suspension was autoclaved at 121oC, 15 mins. MA tests were performed according to a tube agglutination test (2) and modified (3) by using each antigen and serum at 0.1 ml. The antigen and serum were

References
1. 2. 3. Chansiripornchai, N. 2007. J. Thai Vet. Med. Assoc. 58(1): 12-20. Alexander, A.M. and Soltys, M.A. 1973. J. Comp. Path. 83: 191-198. Pholamphisathit, S. 1987. Thai. J. Vet. Med. 17(2): 201-206.

Table 1 Antibody titers against FC by Microplate agglutination (MA) test and protective level after challenge
Group 1 Ages (weeks) 3 4 5 6 7 8 9 10 11 12 13 14 MA titer (Mean SD) 0.74 0.87 2 18.55 11.4 7.6 5.41 22.35 43.5 46 alive/ total ducks 20/20 20/20 20/20 20/20 20/20 20/20 17/20 17/20 16/20 16/20 Group 2 MA titer (Mean SD) 0.74 1.27 0.87 1.29 18.2 15.8 9.2 3.79 25.5 27.1 29.6 18.3 32 17.5 22.93 18.85 20 11.2 alive/ total ducks 16/16 16/16 16/16 16/16 16/16 16/16 16/16 16/16 16/16 16/16 14/16 14/16 Group 3 MA titer (Mean SD) 0.74 0.87 0.6 4.67 4.6 3 alive/ total ducks 10/10 10/10 10/10 10/10 10/10 10/10 0/10

1.27 1.29 1.59 12.44 5.39 3.50 3.37 14.77 22.24 27.94

1.27 1.29 0.97 4.38 4.12 1.04