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Study on the Efficacy of Thermostable Ppr Vaccine, Its Residual Pathogenecity and Molecular Characterization.

M. Giasuddin
Animal Health Research division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

M. M. Rahman
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202.

R. parvin
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202.

A. R. Bhuiyan
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202.

M. J. F. A Taimur
Animal Health Research division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

E. H. Chowdhury
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202.

M. H. Al-Faruque
Animal Health Research division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

M. R. Islam
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202.

Abstract/ Executive Summary:

To determine the efficacy of thermostable PPR vaccine produced by BLRI was preserved at gradient temperature 25?, 30?, 35?, and 40?, for the time of periods of 7 days and 14 days. A total of 90 goats of both sexes, ages between 6 to 12 months, were selected for this experiment. Blood samples were collected from all the groups at pre-vaccination of all vaccinated groups were found positive. However, the percentage of positive slightly decreased in all groups after 180 day post vaccination. No positive sample was found in case of control group. On cell culture, the cytopathic effect (CPE) was visible from 48 hours of post infection by the vaccine virus preserved for 7 days on the shelf, CPE in Vero cells showed initial cell rounding followed by aggregation and syncytial development.

Keywords: Efficacy, Thermostable, PPR vaccine, Residual pathogenecity, Molecular characterization.

Location: Animal Health Research division, Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

Start Date: 01-07- 2009

End Date: 30-06-2010

Program Area: Animal Health and Management

Commodity: Goat

Objectives:

1. To study the shelf life and the efficacy of thermostable PPR vaccine in different temperatures. 2. Molecular characterization of thermostable PPR vaccine virus.

Methodology:

1. Production of thermostable PPR vaccine: A thermostable PPR vaccine has already been developed that could withstand the normal the normal environmental temperature up to 14 days without maintaining any cool chain and it was found most effective to control PPR in goat after both intensive and extensive trials. A special type of programmable Freeze Dryer has been purchased recently by BLRI for production and further research works on the thermostable PPR vaccine. This year (2007-08) thermostable PPR vaccine will be produced and its efficacy and potency trials will be carried out in different field levels. For this, goats in different districts will be vaccinated with the thermostable PPR vaccine that will be transported in the field without maintaining any type of cool-chain. 12 upazilla of 12 districts of six divisions (2 districts from each six divisions) will be included under the study. 2. Testing of efficacy at different temperatures and shelf life: Since environmental temperature in Bangladesh fluctuates with seasons, the vaccines will be kept in different temperatures (20C - 40C) in humid conditions for several days and will be placed in the shelf. At least 50 goats vaccinated for each group. At prior vaccination, 15- and six months days post vaccination, representative serum samples will be collected and analyzed by c-ELISA. Antibody responses in goats from different places will be determined as PI values (PI values> 50 = positive0 and will be compared between different groups and places. 3. Molecular characterization of PPR virus Extraction of Nucleic Acid: The Nucleic Acid from vero cells infected by using RT-PCR method. Reverse Transcription-Polymerase Chain Reaction (RT-PCR): Fragments for all six structural genes of the PPR virus genome will be considered. Specific gene sequence of the vaccine virus will be amplified by RT-PCR using specific primers or the primers will be designed based on the published sequence of the PPR virus genome. The suitable RT-PCR will be used for the amplification.Restriction Fragment Length Polymorphism (Optional): The genome fragment from different vaccine and field isolated amplified by PCR will be treated with different restriction endonuclease enzymes. Vaccine and field virus isolates of the PPR virus will be compared based on the restriction site differences in the genome fragments. Cloning and Sequencing: The genome fragments amplified by PCR will be cloned in TOPO TA-Cloning plasmid vector (PCR2.1, Invitrogen). The recombinant plasmid will be transformed into component E.coli cells (Invitrogen). The insert within the recombinant plasmid will be identified by PCR screening and restriction enzyme analysis. Then the insert within the recombinant plasmid will be sequenced using an automated sequencer. The sequenced data will be analyzed by software supplied with the sequencer or by other software. The nucleotide sequence of the genome fragment from different field and vaccine strains will be compared between themselves, and also with the sequences of other isolates in the GeneBank. A phylogenetic analysis will be carried out to know the evolutionary origin of the Bangladeshi vaccine and field isolates.

Source of Information: BLRI. Proceedings of the Annual Research Review Workshop 2009-2010 Bangladesh Livestock Research Institute, Savar, Dhaka 1341.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Based on these results, it may be concluded that this present vaccine provides only partial protection for a short time. There may be a defect in either seed virus or any other production procedures. It may also be concluded that the thermostable PPR vaccine dept at different environmental temperature (upto 40°C) as long as 14 days able to produce same level of antibody which remain upto 21 days of post vaccination but in comparison with the longevity of protection vaccine preserved for 7 days in better. For molecular characterization, RT-PCT has been standardized for all the six genes (N, P, M, F, H and L gene) of PPR virus.

Knowledge Management: Report/Proceedings