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Prevalence of Foot and Mouth Disease Viruses in Bangladesh and Their Interaction in Bhk-21 Cell Line

M. Rahamat Ullah
Scientific Officer
Poultry Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka-1341, Bangladesh

M. M. Rahman
Bangladesh Agricultural University, Mymensingh

J. Alam
Scientific Officer
Animal Health Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka-1341, Bangladesh

Abstract/ Executive Summary:

A total of 200 Foot and Mouth Disease (FMD) suspected samples collected from different areas of Bangladesh were inoculated both in cell line and suckling mice. Out of the 200 field samples, 170 showed characteristic changes antigen detection in Baby Hamster kidney cell culture (BHK - 21) and 165 killed suckling mice within specified period. Most samples were tested by ELISA for serotyping. The present study demonstrated that three different types of FMD viruses, Type- O, 80%; Type- A, 15%; and Asia- 1, 5% were prevalent in Bangladesh during January 2003 to October 2004. To observe the cell- virus interactions, each isolated serotype was cultured in BHK- 21 cell line and observed for 24 hours. Data were collected at 16 and 24 hours post inoculation. About 80% cells were found round and with pyknotic nuclei incase of serotype- O at 16 hours post inoculation. Where as, serotype- A and Asia-1 produced similar type of lesions, 60% and 70%, respectively in cell line at same interval. After 24 hours post inoculation about 80% cells were washed-out incase of serotype- O followed by 50% and 60% washed-out incase of serotype- A and Asia-1, respectively. From the results of interaction between virus and cell line, it was clear that the predominant O- type FMD virus was able to cause more cellular effect than Asia- 1 and type- A.

Keywords: Prevalence, Foot and Mouth Disease, Interaction in BHK- 21 cell line

Location: Serajgonj, Pabna, Joypurhat, Lalmonirhat, Gaibandha, Kurigram, Dhaka, Comilla, Mymensingh, Khulna, Bagerhat, Barisal, Cox's- Bazar

Start Date: 01-01-2003

End Date: 31-08-2004

Program Area: Pest Management

Commodity: Virus

Objectives:

To determine the prevalence of different serotypes of FMD viruses in Bangladesh and their effect on baby hamster kidney (BHK- 21) cell line.

Methodology:

A total of 200 samples of vesicular fluid, tongue epithelium, gum tissue and tissues from interdigital spaces were collected aseptically in 50% buffered glycerine from the FMD suspected live cattle from different outbreak areas like Serajgonj, Pabna, Joypurhat, Lalmonirhat, Gaibandha, Kurigram, Dhaka, Comilla, Mymensingh, Khulna, Bagerhat, Barisal, Cox's- Bazar of Bangladesh. The tissue samples were then washed by chilled phosphate buffered saline (PBS, PH 7.4) and triturated with sterile pastel and mortar. A twenty percent suspension was prepared from each of the specimens in phosphate buffered saline (PBS) and then centrifuged maintaining 4°C at 5000 rpm for 30 minutes after adding equal volume of chloroform. The supernatant was collected and antibiotic (Gentamicin) @ 0.50 mg/ml was added to prevent bacterial contamination and used as an inoculum for virus isolation. The inocula were inoculated in BHK- 21 cell line and 2- 3 days old suckling mice for virus isolation. After inoculation to culture media of BHK- 21 cell line at a ratio of 1: 10, the cultures were incubated at 37°C for 72 hours. Then the growth of virus was observed by determining cytopathic effects in cell line. After incubation, fluid was collected and centrifuged at 2000 rpm for 30 minutes in refrigerated centrifuge machine to collect sediment of the cells. The supernatant was collected and stored at - 45°C in cryovials for further use. In case of mice, antibiotic treated inocula were inoculated @ 0.2 ml/ mouse through intraperitoneall route and observed for sickness or death for 72 hours. The dead mice were eviscerated and ground with pastle and mortar using sterile sand and a 20% suspension was prepared using PBS. The suspension was then centrifuged and the supernatant was collected for further use. FMD virus positive field samples were used in the identification of the serotypes of virus by Sandwich ELISA using the materials supplied with FMD ELISA kit (Institute for Animal Health, Pirbright). A panel of monoclonal antibodies against different serotypes (Type- O, A, C, Asia- 1, SAT- I, SAT- II, SAT- III) of FMD virus was used for the detection of serotypes of FMD viruses in the cattle population prevailing in Bangladesh. After serotyping, the isolated virus samples were inoculated to BHK- 21 cell line @ 1000 TCID₅₀ (Tissue culture infective dose 50) to determine the pathogenic effects of different FMD viruses. The culture bottles were then kept in an incubator at 37°C and observed at different time intervals (at 16 and 24 hours post inoculation) for cytopathic effects (CPE). The cell lines containing CPE were then fixed, stained and studied for investigation of interactions.

Source of Information: BANGLADESH JOURNAL OF LIVESTOCK RESEARCH, Vol. 12 (1 &2), (Jan - June and June - Dec) 2005, pp. 91-97, ISSN 1022 - 3851.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The study revealed that three different serotypes of FMD virus, type- O; type- A and Asia- 1 were prevalent in Bangladesh during January 2003 to October 2004. About 80% FMD outbreaks were found to be associated with serotype a virus. From the cell- virus interaction, this study also indicated that the O type FMD virus was much more able to cause cellular effects and was more pathogenic than type- A and Asia- 1.

Knowledge Management: Journal