Md. Mostakin Ahamed
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Muhammad Tofazzal Hossain
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Marzia Rahman
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
K. H. M. Nazmul Hussain Nazir
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Mohammad Ferdousur Rahman Khan
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Md. Shafiullah Parvej
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Wahedul Karim Ansari
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Meher Negar Noor-A-Alahi Chiste
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Khaled Bin Amin
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Md. Liakot Hossen
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Sultan Ahmed
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
M. Bahanur Rahman
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.
Molecular characterization, Duck Plague virus, Isolation
Three districts of Bangladesh namely Netrokona, Rajshahi and Mymensingh.
Pest Management
Study area and period: The study was conducted with the samples collected from three districts of Bangladesh namely Netrokona, Rajshahi and Mymensingh during the period of July 2013 through September 2014. Out of these three districts, Netrokona and Mymensingh districts have vast areas of low lands and water bodies, where duck population is significantly high.Samples and methods of sampling: A total of 94 samples comprising of 77 cloacal swabs from suspected ducks, and 17 visceral organs from dead ducks were collected from commercial farms (n=6) and households (n=13) located in the three districts mentioned above, as per the procedure described by Hanaa et al. (2013). The samples were collected aseptically and were placed separately in sterile falcon tubes with proper labeling. The field samples were either processed immediately or stored at -20°C until used. The sample collection from the suspected live birds or experimentation with laboratory animals were performed as per the ethical guidelines set by the Laboratory Animal Unit of the Department of Microbiology and Hygiene, BAU. The samples were collected from the commercial farms and households after taking necessary permission from the owners.
Sterility test and propagation of virus: Antibiotic treated inocula were tested for sterility in fresh blood agar media at 37°C for 24 h . Sterile inocula were then injected through CAM route in 11-13 days old embryonated duck eggs (EDE). After 6-8 days of post-infection (PI), all live EDEs were chilled overnight and allantoic fluid (AF) and CAM were collected. the EDE died earlier was also chilled, and in similar way, the AF and CAMs were collected.
Preparation of hyperimmune sera against DPV: Attenuated Duck Plague Vaccine (FnF®) was inoculated in rabbit with increasing doses for 7 consecutive days (0.1-1 mL, intra-peritoneally). After 14 days of last injection, blood was collected aseptically, and serum was separated, and heat inactivated at 56°C for 30 min. Finally, the serum was stored at -20°C for agar gel immunodiffusion test (AGIT) and passive hemagglutination test (PHA) test, as described by Morrissy et al. (2008).
Detection of duck plague virus by PHA test: The suspension prepared from the CAMs of positive samples were used as the source of virus, which was used for PHA test. All the positive CAM suspensions were found to hemagglutinate the tanned sheep RBC (sRBC) indicating the samples as positive for DPV.
Pathogenicity tests of DPV: Pathogenicity test was done according to the method described by Hanaa et al. (2013). At first, the dELD50 was determined and then ducklings were inoculated with 0.5 mL (105.6 dELD50/mL), and adult ducks were inoculated with 1 mL (105.6 dELD50/mL) of the infectious AF. Pathogenicity test in adult ducks: A small group of adult ducks were intramuscularly inoculated with 1 mL (105.6 dELD50/mL) of DPV, and were observed for 12 days. Few adult ducks were kept separately as control.
Sequencing and phylogenetic analysis: PCR product of partial DNA polymerase gene was sequenced from International Center for Diarrheal Disease Research’ Bangladesh (ICDDR’B). The partially amplified DNA polymerase gene product (446-bp) was sequenced.
Journal of Advanced Veterinary and Animal Research, Vol-2, Issu-3, 2015
Journal