Both nucleotide and amino acid sequences of F protein were individually retrieved from the NCBI GenBank in fasta format, matching those present in viral zone UniProtKB/Swiss-Prot entries using accession numbers. These sequences were edited to equal length and aligned using ClustalW provided in Bioedit software version 7.0.9 (38). Nucleotide similarity calculated using MegAlign Pro (DNASTAR program for life sciences, version 9, Inc., Madison, WI, USA). The waterfowl partial F sequences were aligned separately and converted into protein sequences using Emboss software (EMBL-EBI). Phylogenetic analysis was performed by Bayesian methods in MEGA version X program (39) and or Geneious Prime 2020 with the NJ Kimura 2-parameter method and 1,000 bootstrap replicates. The phylogenetic analysis was done on nucleotides based on the pilot tree proposed by Dimitrov et al. (12) to maintain the tree topology and to ascertain the genotypes of the waterfowl isolates. The same nucleotide sequences were later translated into amino acid sequences in MEGA X program v10.1.8 to compare our waterfowl, poultry isolates, and vaccine (LaSota: AY845400 and I2: AY935499) strains at amino acid levels.

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