## 3 Results  
### Nucleotide Identity Comparisons  
The two regions of the ASaV RNA1 genome, the ORF1 N-proximal region and ORF1 C-proximal region, are located at positions 840-1875 and 4361-5474 (numbers based on antigenome reference isolate acc. no. OU466880), respectively. The pairwise comparisons of 11 samples including E59464, E60765, E62017, E62033, E62787, E62907, E63064, E63065, E63069, E63071, and E63073 for ORF1 N-proximal and E60765, E62017, E62033, E62907, E63064, E63065, E63069, E63071, E63073, E62477, and E62941 for ORF1 C-proximal, along with two isolates from GenBank (accession numbers OU466880 and OU466875), showed nucleotide-level identities of 92.88-100% and 87.62-99.91%, respectively, in the sequenced isolates. In the ORF1 N-proximal region, the highest identity (100%) was recorded between E62907 (Melzower forest in eastern Germany) and isolates E63069 and E63073 (from Jever and Wangerland in the west of Germany). The lowest identity (92.88%) was observed between E62033, sampled from Melzower forest in northeast Germany, and E55270, sampled from Basel, Switzerland. In the ORF1 C-proximal region, the lowest identity (87.62%) was observed between isolate S10 (Hamburg, Germany) and E63064 (Wangerland, Germany). The highest identity (99.91%) was observed between two isolates from Melzower, forest E62477 and E62033. Interestingly, these isolates shared only 89% identity with another isolate from the same location, E62907, indicating genetic variability among the group of isolates collected from the same location (Melzower forest, Germany).  
RNA2 was amplified in 20 isolates, RNA3 in 41 isolates, RNA4 in 26 isolates, and RNA5 in 23 isolates. These sequences were used for pairwise alignment comparisons with available ASaV genome sequences in the GenBank database, including RNA1-RNA5. The GenBank isolates used for alignment included RNA2 (OU466881), RNA3 (MN399721, MN399722, OP501825, OU466877, OU466882), RNA4 (OP501826, OU466878, OU466883), and RNA5 (OU466879, OU466884). The nucleotide identity of 92.75-100%, 91.97-100%, and 90-100% was obtained in pairwise comparisons of almost full-length RNA2 to RNA5 of the sequenced isolated (Supplementary Table S2). RNA4 was the most conserved genomic fragment, while RNA5 and RNA3 were the most divergent genomic segments of ASaV in the studied isolates. The E62015 (Melzower forest, Germany) isolate was the most divergent isolate based on the RNA2. Based on the pairwise sequence identity of RNA3, the lowest identity was observed between E62779 (Kaisheim, Germany) and E62017 (Berlin, Germany). The amino acid's lowest identity (93.1%) was observed between E58051 (_F. ornus_, Hamburg, Germany) and a group of isolates including E62017 (Berlin, Germany), E62037, E62053, E62073, E62900 (Melzower forest, Germany), and PaEV(_Pisum sativum_, Meissen county, Germany, acc. no. MN399722). In the case of RNA4, the lowest sequence identity (95%) was observed between isolates E62900 (Melzower forest, Germany) and OP501826.1 (Castets-en-Dorthe, France). At the deduced amino acid (aa) level, the sequence identity exceeded 99.16%. The lowest observed identity based on the RNA5 was found between isolates E62001 (Hamburg, Germany) and E62231 (_F. ornus_, Berlin, Germany).  
The nucleotide comparisons between the isolates collected from the same location based on the sampling sites, i.e., Berlin, Melzower forest, Hamburg, Wangerland, Jever, Wittmund, Goting, and Kaisheim, showed that the isolates collected from Berlin and Melzower forest had the most divergent isolates.