One-sentence summary of the conclusion 33 We show that the ruminant-associated Influenza D virus has direct transmission capability to 34 humans. 35 36 Abstract 37 Influenza viruses are notorious pathogens that frequently cross the species barrier with often 38 severe consequences for both animal and human health. In 2011, a novel member of the 39 Orthomyxoviridae family, Influenza D virus (IDV), was identified in the respiratory tract of 40 diseased swine. Epidemiological surveys revealed that IDV is distributed worldwide among 41 livestock and that IDV-directed antibodies are detected in humans with occupational exposure 42 to livestock. To identify the transmission capability of IDV to humans, we determined the 43 viral replication kinetics and cell tropism using an in vitro respiratory epithelium model of 44 humans. The inoculation of IDV revealed efficient replication kinetics and apical progeny 45 virus release at different body temperatures. Intriguingly, the replication characteristics of 46 IDV revealed many similarities to the human-associated Influenza C virus, including the cell 47 tropism preference for ciliated cells. Collectively, these results might indicate why IDV-48 directed antibodies are detected among humans with occupational exposure to livestock. 49 50 51After the initial discovery of Influenza D virus (IDV) in 2011, among swine with Influenza-52 like symptoms, knowledge about this new genus in the family of Orthomyxoviridae is 53 increasing (1, 2). Epidemiological studies have shown that the virus has a worldwide 54 distribution, whereby at least two distinct genetic lineages are cocirculating and reassorting 55 (3-10). Because of the high seroprevalence, cattle is the proposed natural reservoir of IDV, in 56 which IDV causes mild respiratory disease symptoms (11). In addition to cattle, IDV-specific 57 antibodies have been detected in swine, feral swine, equine, ovine, caprine and camelid 58 species, suggesting a broad host tropism for IDV (3, 4, 9, 12, 13). However, the most striking 59 observation is the detection of IDV-directed antibodies among humans with occupational 60 exposure to livestock (14).
61There are several indicators that IDV has a zoonotic potential. For instance, the 62 utilization of the 9-O-acetyl-N-acetylneuraminic acid as a receptor determinant, that allows 63 the hemaglutinin esterase fusion (HEF) glycoprotein of IDV to bind the luminal surface of the 64 human respiratory epithelium (1). Interestingly, the utilization of this receptor is also 65 described for the closely related, human associated Influenza C virus (ICV) (15, 16). 66 Furthermore, the detection of IDV-directed antibodies among humans with occupational 67 exposure to livestock and the molecular detection of IDV in a nasopharyngeal wash of a field 68 worker with close contact to livestock indicates that cross species transmission occurs (14, 69 17). However, thus far, there is no indication of wide spread prevalence among the general 70 population although the virus has been detect...