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Experimental Preparation of Vaccines for Immunization of Chickens Against Fowl Pox-i. Isolation, Characterization and Experimental Immunization.

M.M. AMIN
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh

A.B. SIDDIQUE
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh

M.HARUN-AL-RASHID
Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

Pigeon pox virus (PPV) were isolated from 40 out of 50 clinical specimens of pock lesion materials collected from individual pigeons naturally infected with pigeon pox virus. The isolates were obtained through growth of virus via chorio-allantoic memberane (CAM) of 12 days old embryonated chicken eggs. The field isolates of PPV were characterized by serum neutralization test (SNT) using homologous serum. The embryo infectivity dose fifty (EID₅₀) titres of selected field isolates were also determined by inoculation of embryonated chicken eggs via chorio-allantoic membrane (CAM). The end point of EID₅₀ were between 10^-6 and 10^-7 in equal numbers of 40 field isolates of PPV. Preliminary investigation of dose response in 3-5 weeks of age were conducted via two routes of inoculation. It was observed that a "take reaction" due to infection of the virus appeared within 7 days of post- vaccination period. Further, the size and number of nodules formed decreased as the dilution of viral suspension increased.

Keywords: Scabs, Pock lesions, EID50, Serum neutralization test

Location: Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Chicken

Objectives:

Preparation of vaccines with PPV control fowl pox in chicks.

Methodology:

Collection of specimens: Specimens such as scabs appearing on different parts of pigeon clinically infected with pox virus were collected aseptically from different localities of Dhaka, Mymensingh and Narayangonj districts. These were kept in ice-flasks or 50% buffered glycerine and carried to the laboratory. Isolation of field virus: For the isolation of virus each sample was macerated individually with sterile pestle and mortar and suspended in phosphate buffer saline (PBS). The suspension was then centrifuged to remove the coarse particle. Supernatant was separated, treated with antibiotics and then inoculated in 12 days-old embryonated chicken eggs via chorioallantioc membrane (CAM) route at the rate of 0.1 ml per embryos. Five embryos were inoculated for each specimen. The embryos were inoubated at 37°C and examined after 5-6 days of inoculation when CAM showing pock lesions due to growth of virus was considered positive. These were collected in sterile vials or test tubes. Samples which showed no pock lesions in CAM after 2-3 blind passages were considered negative. Storage of PPV isolates: The isolates grown on CAM were kept stored at - 20°C until used. Characterization of virus isolates: These were performed by serum neutralization test (SNT) using antiserum against PPV. Serial two fold dilution ranging from 1:2 to 1: 1 024 of the serum raised in chicken against PPV were made in PBS. It was then mixed with standard volume (0.5 ml) of isolate (0.1 ml virus preparation containing 100 EID titre). The virus serum mixtures was incubated for an hour at room temperature and then inoculated into 12 days-old chicken embryo through CAM route. The inoculated embryos along with uninoculated control of embryonated eggs were incubated for 5-6 days and then examined for the formation of pocks on CAM. Determination of dose response: For this 5 (Five) selected field isolates of PPV grown earlier on CAM and kept stored at -25°C were macerated and suspended in PBS individually. Supernatant collected after centrifugation were diluted upto 1 O^-5 . Each dilution of virus suspension was inoculated via subcutaneously or by feather follicle route at the rate of 0.1 ml per bird. Thus a total of 25 chicks of chickens aged 3-5 weeks were used for each experimental virus isolate. The birds were observed for 21 days during which the serum samples were obtained from individual bird at 14 and 21 days post-vaccination period.

Source of Information: BAU Res. Prog. 7: 401-404; 1993

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Investigation on the dose response indicated that a concentration of 50 EID₅₀ titres of virus exhibited "take reaction" within seven days of inoculation via either subcutaneous or feather follicle routes. During the course of this study it was also observed that the size and number of nodules formed due to "take reaction" decreased as the virus dilution increased.

Knowledge Management: Report/Proceedings