Bovine papillomaviruses (BPV) are members of the family Papillomaviridae. Papillomavirusesare usually species-specific and epitheliotropic. Delta-BPVs are an exception to this rule in that theycan also infect fibroblasts and non-bovid ungulates. Cell cultures are essential for performing invitro studies, analysis of virus biology, vaccine production, tissue engineering and toxicity testing.In this context, cell line contamination constitutes a significant problem. In this study, variouscell lines (n=27) were assessed for potential BPV contamination. To this aim, DNA was extractedfrom cell cultures and then screened for the presence of papillomavirus L1 capsid gene DNAusing a consensus polymerase chain reaction (PCR) system. Immunofluorescence (IF) stainingwas used for viral protein detection. Intriguingly, one cell line derived from equine dermis testedpositive by PCR and subsequent IF staining for L1. Amplicon sequencing followed by computedL1 DNA sequence alignment led to the identification of a new putative BPV type, revealing thehighest identities with Delta-BPV types 1 (76%) and 2 (73%). To the authors’ knowledge, thisis the first report on the presence of a putative BPV as a viral contaminant in cell cultures thatpossibly represents an unknown Delta-BPV.