With this study, we were able to describe the detection and characterization of the first three paramyxoviruses in insectivorous bats. The genetic distance between these three novel paramyxoviruses and the closest related member known is higher than that of members within other paramyxovirus genera, suggesting that all three viruses might be considered as unassigned paramyxoviruses. Thus the two viruses BatPV/Myo.mys/E20/09 and BatPV/ Figure 1 . Improved detection sensitivity after Taguchi optimization of the Paramyxovirinae subfamily-specific PCR [21] . Gel electrophoresis of amplification products of the second round seminested PCR using 10-fold serial dilutions (10 0 to 10 24 ) of a cDNAsample (kidney of sample E20/09). (A) PCR protocol adopted for twostep PCR as previously published [21] using the pan-PAR-F1/PAR-R primer pair (1st run) and the pan-PAR-F2/PAR-R primer pair (2nd run). (B) Optimized protocol using the pan-PAR-F1/PAR-R primer pair (1st run) and the pan-PAR-F2/PAR-R primer pair (2nd run). doi:10.1371/journal.pone.0038688.g001 Figure 2 . Phylogenetic analysis of the partial L-gene sequence obtained from PCR fragments after Pan-Paramyxovirinae-PCR with PAR primers (529 bp) [21] . The revealed gap-free alignment was used to generate a phylogenetic tree of the novel bat paramyxoviruses (red) concordant with representatives from all known genera of paramyxoviruses with MrBayes. Posterior probability rates are given next to the tree nodes.