Figure legends
Figure 1. Detection rates of OaPV single infections. Differences were significant after Cochran-Armitage test (p <0.05).
Figure 2. (A) Electrophoresis of PCR products for evaluating OaPV1 E5, OaPV2 L1, OaPV3 E7, and OaPV4 E6 DNA in bovine PBMC samples. MW: DNA molecular weight marker (100 bp). C: PCR negative control. (B), (C), (D), and (E):100% identity between the sequences of the amplicons OaPV1 Seq, OaPV2 Seq, OaPV3 Seq, and OaPV4 Seq, and the sequences reported in GenBank, respectively (accession number: U83594.1; U83595.1; NC_038516.1; and KX954121.1).
Figure 3. One Step ddPCR. The rain plots for the OaPV1 E5, OaPV2 L1, OaPV3 E6 and E7, and OaPV4 E6 are shown. Representative samples have positive droplets at the same amplitude as the positive control for each OaPVs.
Figure 4. (A) Electrophoresis of RT-PCR products to evaluate OaPV1 E5, OaPV2 L1, OaPV3 E6 and E7, and OaPV4 E6 mRNA expression in PBMC samples. MW: DNA molecular weight marker (100bp); C+: PCR positive control. PCR was performed on PBMC samples with (RT+) and without (RT-) the addition of reverse transcriptase to the reaction mixture, using the same amount of RNA. (B) The amplicon sequences showed 100% identity with the respective sequences reported in S1 Fig. Furthermore, 100% identity between the OaPV3 E6 cDNA amplicon and the OaPV3 E6 sequence reported in GenBank (accession number: NC_038516.1) is shown.
Figure 1 SuppInfo. Detection and quantification of OaPV DNA using ddPCR. The Bio Rad system quantified DNA copies per μL. Bov = bovine; N = negative
Figure 2 SuppInfo. Detection and quantification of mRNA using one-step reverse transcription (RT)-ddPCR. The Bio Rad system quantified mRNA copies per μL. Bov = bovine; N = negative
Figure 3 SuppInfo. (A) . Representative positive samples and a positive control for the OaPV1 E5 and OaPV2 L1 assessed by ddPCR in grass hay and maize silage samples. Positive samples have droplets at the same amplitude as the positive control. Amplitude of OaPV1 E5 ranged from 1.500 to 5.500; amplitude of OaPV2 L1 ranged from 1.500 to 11.500. (B). PCR products of the above samples. MW: DNA molecular weight marker (100bp). C+: PCR positive control; C-: PCR negative control. The amplicon sequences showed 100% identity with the respective sequences reported in GenBank (see Figure 2).
Table 1 shows the primers and probes used for the detection and quantification of OaPVs.
Table 2- Genotype coinfections evaluated by ddPCR with number and related percentages of their combination are shown.