Recent strains of influenza D virus create a new genetic cluster for European strains.

Bovine respiratory diseases (BRD) are one of the significant health problems for cattle breeding industry. Influenza D virus (IDV) alone or in combination with other respiratory pathogens plays a role in BRD. According to the IDV-HEF gene region, phylogenetic analyzes revealed five lineages: D/OK, D/660, D/Yama2016, D/Yama2019, and D/CA2019, so far. In this study, despite no success in virus isolation, the presence of IDV was investigated by RT-PCR (partial HEF gene region) in 219 nasal swab samples collected from cattle with BRD between 2012 and 2021. The presence of IDV was demonstrated in two samples, and genome characterization data of the IDV sequences both in the partial and complete HEF gene regions showed that one of the obtained sequences (D/bovine/Turkey-Bursa/ET-138/2021) was in the lineage D/Yama2019 while the other (D/bovine/Turkey-Bursa/ET-130/2013) created a new lineage tentatively called D/Bursa2013 as including few partial IDV sequences reported in Europe. Two nucleotide substitutions (nt252A→G, nt299T→C) were typically characterized for the tentative lineage D/Bursa2013, one of which also leads to a unique amino acid change at position aa100 (V→A). When the amino acid differences between the lineages were evaluated, amino acid substitution changes were detected in four regions [aa12 (Alanine→Aspartic acid), aa19 (Glycine→Arginine), aa22 (Proline→Serine), and aa110 (Aspargine→Arginine)] of the D/Yama2019 lineage, unlike the other lineages. Considering the most common D/OK lineage in Europe, many nucleotide substitutions were shown between D/OK and D/Bursa2013. Accordingly, aminoacid substitutions were observed in aa27 (Threonine→Asparagine) and aa100 (Valine→Alanine) in the D/bovine/Turkey-Bursa/ET-138/2021 sequence. Study results describe the circulation of D/Yama2019 and D/Bursa2013 (new lineage) in Turkey. Expansion of new strains seems possible due to the high mutation rate of influenza viruses. It is important to understand the development of IDV with comprehensive characterization studies.