ABSTRACT
Objective: The research aimed to isolate, adapt to cell culture, and characterize the lumpy skin disease virus (LSDV) from clinically infected cattle in Bangladesh. Materials and Methods: From September 2019 to June 2020, 37 skin nodules and skin swabs were aseptically collected from afflicted cattle in the outbreak regions of Jhenaidah and Kishoreganj in Bangladesh. The LSDV was isolated from embryonated specific pathogen-free (SPF) chicken eggs along the chorioallantoic membrane (CAM) route and the Vero cell line after several blind passages. The viral attachment protein was targeted for molecular detection using polymerase chain reactions (PCR). For phylogenetic analysis, PCR-positive products were partially sequenced. Results: The virus was evident in the cell line, showed cytopathic effects after the 13 blind passage, and on the CAM of SPF chicken eggs, exhibited thickening of the CAM with pock-like lesions. A total of 12 samples (32.43%) tested positive for LSDV by PCR. Phylogenetic analysis of the present isolates (accession numbers MN792649 and MN792650) revealed 100% similarity with strains from India (MN295064), Kenya (AF325528, MN072619, KX683219), Greece (KY829023), Serbia (KY702007), and Kazakhstan (MN642592); moreover, 99.43% to 100% similarity to the sheep pox virus. Conclusion: Partially sequenced LSDV was developed as a vaccine seed and was first isolated in Bangladesh and characterized at the molecular level.
ABSTRACT
OBJECTIVES: Migratory birds play a major role in the transmission of pathogens globally, but still their role in the transmission of fungi in Bangladesh is not known. The present study was carried out for the isolation and molecular detection of fungi including Aspergillus from migratory birds traveling to Bangladesh. MATERIALS AND METHODS: A total of 50 fecal samples were collected from BaojaniBaor, Magura, and areas close to Jahangirnagar University, Savar. The isolation of fungus was based on culture on Potato Dextrose Agar (PDA), followed by staining, morphology, and molecular detection using polymerase chain reaction (PCR). RESULTS: Among 50 samples, 40 showed positive for fungal growth on PDA, of which 30 yield only yeast-like colonies, five only molds, and five yielded both yeast and molds. The isolated molds produced various pigmented colonies, namely, black, whitish, grayish, olive green, and yellow. Among 10 molds, six were confirmed as fungi by PCR using genus-specific primers such as ITS1 and ITS4. Later, of these six fungi, five were confirmed as Aspergillus by PCR with primers such as ASAP1 and ASAP2 specific for Aspergillus genus. Therefore, the overall occurrence of Aspergillus was 10% (5/50). PCR specific for Aspergillus fumigatus and Aspergillus niger failed to produce specific PCR amplicon, suggesting that the isolated Aspergillus belongs to other groups. CONCLUSION: This is the first report describing the isolation and molecular detection of Aspergillus from fecal samples of migratory birds in Bangladesh. The present findings confirm that migratory birds are potential source for Aspergillus and other fungus in Bangladesh.
