Markus Hodal Drag 1,2 *, Christina Hvilsom 1, Louise Ladefoged Poulsen 2, Stamatios Alan Tahas 1, Carolyn Cray 3, Mads Frost Bertelsen 1 and Anders Miki Bojesen 2

1Copenhagen Zoo, 2000 Frederiksberg, Denmark; 2Department of Veterinary and Animal Sciences, University of Copenhagen, 1870 Frederiksberg C, Denmark; 3Division of Comparative Pathology, University of Miami Miller School of Medicine, Miami, FL 33136, USA.

Avian aspergillosis (AA) is an important and potentially fatal respiratory fungal infection affecting a broad range of wild and domestic birds, both in natural environments, in zoological gardens and nature reserves, and in agriculture. The suboptimal sensitivity and specificity of current AA diagnostics have stressed the need for new techniques that enable early subclinical detection. This project seeks to develop a novel diagnostic panel using a modern omics-based approach, with a focus on Oxford Nanopore sequencing, high resolution MS/MS, and ultra-sensitive antibody detection systems. Using samples from a newly established chicken model infected with spores from Aspergillus fumigatus, the panel will evaluate: i) changes in epigenetic signatures of circulating cell-free DNA (cfDNA) to reveal infection-mediated cell death, ii) the presence of microbial cfDNA and mycotoxins, and iii) changes in specific acute phase proteins and antigens. The combination of these new markers, which are available in simple routine blood draws, will allow unprecedented early detection of AA and enable efficient treatment and removal of source of exposure before infections turn severe.