LAUSR

Significance Emerging respiratory pathogens pose significant public health threats as a result of their potential for rapid global spread via multiple non-mutually exclusive modes of transmission. The relative significance of contact, droplet, and airborne transmission for many respiratory pathogens remains a knowledge gap, and better understanding is essential for developing evidence-based measures for effective infection control. Here, we describe and evaluate a transmission chamber that separates virus-laden particles in air by size to study airborne particles that mediate influenza transmission in ferrets. Our results provide direct experimental evidence of influenza transmission via droplets and fine droplet nuclei, albeit at different efficiency. This transmission device can also be applied to elucidate the mode of transmission of other respiratory pathogens. Epidemics and pandemics of influenza are characterized by rapid global spread mediated by non-mutually exclusive transmission modes. The relative significance between contact, droplet, and airborne transmission is yet to be defined, a knowledge gap for implementing evidence-based infection control measures. We devised a transmission chamber that separates virus-laden particles by size and determined the particle sizes mediating transmission of influenza among ferrets through the air. Ferret-to-ferret transmission was mediated by airborne particles larger than 1.5 µm, consistent with the quantity and size of virus-laden particles released by the donors. Onward transmission by donors was most efficient before fever onset and may continue for 5 days after inoculation. Multiple virus gene segments enhanced the transmissibility of a swine influenza virus among ferrets by increasing the release of virus-laden particles into the air. We provide direct experimental evidence of influenza transmission via droplets and fine droplet nuclei, albeit at different efficiency.