Knowledge of host associations of blood-feeding vectors may afford insights into managing disease systems and protecting public health. However, the ability of methods to distinguish bloodmeal sources varies widely. We used two methodsSanger sequencing and amplicon deep sequencingto target a 228 bp region of the vertebrate CYTB gene and determine hosts fed upon by triatomines (n = 115) collected primarily in Texas, USA. Direct sanger sequencing of PCR amplicons was successful for 36 samples (31%). Sanger sequencing revealed 15 distinct host species, which included humans, domestic animals (
Canis lupus familiaris, Ovis aries, Gallus gallus, Bos taurus, Felis catus, and Capra hircus), wildlife ( Rattus rattus, Bufo nebulifer, Sciurus carolinensis, Sciurus niger, Odocoileus virginianus), and captive animals ( Panthera tigris, Colobusspp., Chelonoidis carbonarius). Samples sequenced by the Sanger method were also subjected to Illumina MiSeq amplicon deep sequencing. The amplicon deep sequencing results (average of 302,080 usable reads per sample) replicated the host community revealed using Sanger sequencing, and detected additional hosts in five triatomines (13.9%), including two additional blood sources ( Procyon lotor, Bassariscus astutus). Up to four bloodmeal sources were detected in a single triatomine ( Bufo nebulifer, Homo sapiens, Canis lupus familiaris, and Sciurus carolinensis). Enhanced understanding of vector-host-parasite networks may allow for integrated vector management programs focusing on highly-utilized and highly-infected host species.