Comparison Of Four Anti-Avian IgY Secondary Antibodies Used In Western Blot And Dot-Blot ELISA To Detect Avian Bornavirus Antibodies In Four Different Bird Species

Escandon, Paulina; Heatley, J Jill; Berghman, Luc R; Tizard, Ian; Musser, Jeffrey MB

The experimental protocol was reviewed and approved by the Texas A&M University Bio-safety and Animal Use Committees (IACUC 2016-0274, 2014-0010, 2016-0054 and 2017-0217), which, complies with the guidelines included in the National Research Council of the National Academies' publication Guide for the Care and Use of Laboratory Animals, 8th edition. Blood samples equivalent to 1% body weight were collected by jugular venipuncture. Blood was collected into lithium heparin tubes and stored at 4°C prior to centrifugation (5000 x g, 10 mins). Plasma was collected and stored at −20°C prior to analysis. Two Blue and gold macaws, one Cockatiel, one Monk parakeet and two Mallards were sampled. Blue and gold macaw blood samples were collected from one bird naturally infected with ABV that routinely tested positive for ABV mRNA by RT-PCR in its urofeces and from one bird that was not known to be exposed to ABV and routinely tested negative by RT-PCR. Cockatiel and Monk parakeet blood samples were collected prior to and following experimental infection; the birds were tested routinely by RT-PCR. The Mallard samples were collected from an experimentally infected, RT-PCR positive Mallard and from a non-infected, RT-PCR negative Mallard.