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Evaluation of Alum Precipitated Formalin Killed Fowl Cholera Vaccines with Their Immunologic Responses in Ducks

M.A. Islam
Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

M.A. Samad
Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

M.B. Rahman
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

The alum precipitated formalin killed fowl cholera vaccines (FCV) are locally produced from the Livestock Research Institute (LRI), Mohakhali, Dhaka and Bangladesh Agricultural University, Mymensingh which are being used to control fowl cholera in chickens and ducks in Bangladesh. Efficacy of these two vaccines has been evaluated mostly in chickens but reports on ducks are very limited. Four weeks old 50 Jinding breed of ducks were used to evaluate the efficacy and immune responses of fowl cholera vaccines during the period from October 2002 to March 2003. These 50 ducks were divided into three groups (A = 20, B = 20 & C = 10 ducks) and each duck of group A was inoculated with FCV (LRI) @ 0.5 ml subcutaneously at the age of 8 weeks, and each duck of group B received FCV (BAU) @ 1.0 ml intramuscularly at the age of 12 weeks as manufacturer instruction, whereas ducks of group C served as unvaccinated control. Each duck of group A and B was also injected booster dose after two weeks of primary vaccination with their respective FCV. Each duck of all the three groups (A, B and C) was challenged after two weeks of post-booster vaccination with 1.0 ml inoculum containing 5.4×10⁶CFU of virulent Pasteurella multocida intramuscularly. The results of challenged experiment revealed that one (5.0%) duck of group A, two (10.0%) ducks of group B died within 2 to 3 days of post-challenged, whereas 10 (100%) unvaccinated control ducks of group C died within 1 to 3 days of post-challenged. Therefore, the FCV® (LRI) conferred protection to 95% and FCV (BAU) conferred protection to 90% of vaccinated birds against challenged infection after two weeks of booster vaccination. The mean values of Total leukocytic count (TLC), Total serum protein (TSP) and Passive haemagglutination assay (PHA) antibody titre of ducks in both the groups A and B were found significantly (p < 0.01) increased at two weeks of post-primary and two weeks of post-booster vaccination, and also two weeks of post-challenged infection in comparison to the respective prevaccination values. These results indicate that the FCV of LRI induced comparatively higher TLC, TSP and PHA antibody titre than FCV of BAU in ducks. These results showed that the locally prepared fowl cholera vaccines induced sufficient cellular and humoral immune responses which resulted satisfactory level of protection against duck cholera and therefore both the locally prepared FCV could be recommended to control duck cholera under filed conditions in Bangladesh.

Keywords: Fowl cholera, Immunization, Duck, Immune response, Protection

Location: Livestock Research Institute (LRI), Mohakhali, Dhaka and Bangladesh Agricultural University, Mymensingh

Start Date: 00-10-2002

End Date: 00-03-2003

Program Area: Animal Health and Management

Commodity: Duck

Objectives:

To determine the immune response with comparative efficacy of locally prepared two fowl cholera vaccines in ducks.

Methodology:

Four-week-old 50 Jinding breed of healthy ducks of either sex with no previous history either vaccination or of fowl cholera infection were purchased from the Government Duck Farm, Kishoregonj. These ducks were divided into three main groups (A, B and C) of which groups A and B consisting of 20 ducks in each, whereas group C consisting of 10 ducks. These three groups of ducks were maintained separately with intensive care and adequate commercial feed (Quality Feeds Ltd., Dhaka) and water supply throughout the experiment. In addition to general feed and water supply, vitamin-mineral premix (Megavit®, Novartis, Bangladesh Ltd., Dhaka) was also supplied in the drinking water daily. Alum precipitated formalin killed fowl cholera vaccine prepared by the Poultry Biologic Unit (BAU), Mymensingh was purchased directly from the manufacturing unit, and fowl cholera vaccine manufactured by the (LRI) Mohakhali was obtained from the district veterinary hospital, Mymensingh were used for this study. Two doses of fowl cholera vaccines were administered at interval of 14 days as per manufacturer immunization of ducks. Each duck of group A (n = 20) was primarily vaccinated at the age of 8 weeks with fowl cholera vaccine prepared by LRI @ 0.5 ml SC followed by booster vaccination with same vaccine at the age of 10 weeks of age. Primary vaccination with fowl cholera vaccine prepared by BAU was made in each of the duck of group B (n =20) at the age of 12 weeks @ 1.0 ml intramuscularly with same vaccine followed by booster vaccination with at the age of 14 weeks. Ducks of group C (n=10) served as unvaccinated and challenged control. The locally isolated virulent Pasteurella multocida serotype I (Strain X-73) maintained in the laboratory was used as challenge organisms. Each of the bird of all the three experimental groups (A, B and C) was challenged intramuscularly with 1.0 ml inoculum contained 5.4 x 10⁶ CFU. All the challenged birds were observed observed daily for two weeks to detect the signs and symptoms, morbidity and mortality by the challenged inoculum. Blood was collected from the wing vein of each of the experimentally vaccinated duck of groups A and B in two separate test tubes. One contained double oxalate as anticoagulant which is used for total leukocytic count and other one without adding any anticoagulant which is used for separation of serum to determine the antibody titre and total serum protein. Blood samples were collected from the ducks at pre-vaccination, two weeks of post-primary vaccination, two weeks of post-booster vaccination and two weeks of post-challenge. Sera were separated from the blood collected without adding any anticoagulant and stored at -20^oC until tested. Oxalated veinous blood taken up to 0.5 mark of a RBC diluting pipette and then drawn the diluting fluid up to the 101 mark, thus made a 1:200 dilution and mixed well with 8 knot method. First one or two drops discarded and then pour of one drop on counting chamber with cover slip and counted the leukocytes on the 9 large squares having a total area of 9 sq.mm and counted the average leukocyte, multiplied by 2000 to gave the TLC in 1 sq. mm of blood. The passive haemagglutination assay (PHA) first was used to determine the antibody titre of ducks immunized with fowl cholera vaccines according to the method described by Tripathy et al. (1970) with slight modification. Tripathy et al. (1970) described the PHA in fowl pox virus with PBS pH 6.4, 1:25000 tannic acid solution, Na₂HPO₄•12H₂O 0.15 M, KH₂PO₂•2H₂O 0.15 M and sensitized RBC 0.5% whereas pH 7.2, 1:20000, 0.2M, 0.2M and 0.7% were used in this study respectively. The ducks which were died at post-challenged were necropsied as soon as they were found dead. In addition three ducks in each the three challenged groups (A, B, C) were randomly selected and sacrificed at 14 days of post challenged. Swabs taken from liver and heart were streaked onto blood agar plates, which were incubated at 37⁰C for 24 hours for the growth of organisms. The positive cases showing growth of P.multocida were confirmed by the standard bacteriological procedure.

Source of Information: International Journal of Poultry Science 3 (2): 140-143, 2004

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Achievement/Findings:

These results showed that the locally prepared fowl cholera vaccines induced sufficient cellular and humoral immune responses which resulted satisfactory level of protection against duck cholera. Therefore both the locally prepared FCV could be recommended to control duck cholera under filed conditions in Bangladesh.

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