Viral genomic sequence isolation and identification.

Viral genomic sequences were identified as part of a previous study (52). Briefly, bats were collected with harp traps at an abandoned railroad tunnel near Little Orleans, MD, and fresh guano was collected. Samples were stored on ice until processing. The BtCalV/A10 capsid sequence was isolated from Perimyotis subflavus (tri-colored bat) using the sequence-independent single-primer amplification technique and 454 next-generation sequencing platform. De novo contigs were assembled using three programs: Codon Code Aligner, Geneious, and DNAStar. Viral amino acid sequences were created using Vector NTI. Assembled contigs were analyzed using the basic local alignment search tool (BLAST) from the National Center for Biotechnology Information (NCBI) (87). BLAST searches were conducted at the amino acid level using the protein-protein BLAST (blastp) function to query nonredundant protein sequences.