Blood Parasite Infection Data from Blue-winged Teal, Canada (Alberta, Saskatchewan) and USA (Texas, Louisiana), 2012-2013

Blue-winged teal were live-captured during March 2012–2013 in Texas and Louisiana and during August 2012–2013 in Alberta and Saskatchewan. Captures were conducted using rocket nets in Texas and Louisiana and standard swim-in bait traps in Alberta and Saskatchewan. Birds were aged and sexed via plumage. All birds sampled in March were considered adults as they had molted into nuptial plumage by the time of capture. A small blood sample (<1 % of body weight) was collected from each bird via jugular venipuncture and a subsample (<0.25 ml) stored in Longmire (Texas and Louisiana) or Queen’s (Alberta and Saskatchewan) buffer for use in this project. Blood samples were held at ambient temperature during sample collection periods and then frozen prior to analysis.

Blood parasites were detected using molecular methods. DNA was extracted from samples using the DNeasy Blood and Tissue Kit (Qiagen, Valencia, CA) and screened for hematozoa using nested PCR protocol. A minimum of one negative control was incorporated into PCR reactions for every 24 wells. Reactions were conducted in eight-well strip tubes with individual caps that remained closed, except while loading template and reagents, to prevent cross contamination. Amplicons were visualized on 0.8% agarose gels stained with Gel Red Nucleic Acid Gel Stain 10,000× in DMSO (Biotium, Hayward, CA). All samples were screened twice for hematozoa by nested PCR. A 479-base pair (bp) cytochrome b (cyt b) mitochondrial DNA (mtDNA) target fragment was sequenced for all samples that had amplicons present on agarose gels. PCR products were treated with ExoSap-IT (USB Inc., Cleveland, OH) per the manufacturer’s instructions and not otherwise purified prior to sequencing. Sequencing was performed with identical primers used for PCR and with BigDye Terminator version 3.1 mix (Applied Biosystems, Foster City, CA) on an Applied Biosystems 3730xl automated DNA sequencer (Applied Biosystems, Foster City, CA). Sequence data were cleaned and edited using Sequencher version 5.1 (Gene Codes Corp., Ann Arbor, MI). Infections were assigned to genera (Leucocytozoon, Haemoproteus, or Plasmodium) using the nucleotide BLAST (Basic Local Alignment Search Tool) function available through the National Center for Biotechnology Information (NCBI). Assignment was based on the top NCBI BLAST result with a minimum identity score of at least 90%. A sample was considered as positive for blood parasite infection in any given PCR run if it resulted in double-stranded, target mtDNA that was verified through genetic sequencing and successfully assigned to one of three parasite genera using BLAST. Genetic characterization of resultant sequences were previously summarized. Single-stranded sequences or products that could not be assigned via BLAST were considered as negative to reduce the possible occurrence of false positives.