Simple SummaryAcute gastroenteritis (AGE) is a very common illness and a significant cause of morbidity and mortality in cats less than one year of age. Along with known viruses historically recognized as primary agents of AGE, in the last years deep sequencing technologies have identified a variety of novel viruses. In this study, we explored the feline enteric virome from cats with or without diarrhoea, unveiling a wide diversity of viruses.AbstractViruses are a major cause of acute gastroenteritis (AGE) in cats, chiefly in younger animals. Enteric specimens collected from 29 cats with acute enteritis and 33 non-diarrhoeic cats were screened in PCRs and reverse transcription (RT) PCR for a large panel of enteric viruses, including also orphan viruses of recent identification. At least one viral species, including feline panleukopenia virus (FPV), feline enteric coronavirus (FCoV), feline chaphamaparvovirus, calicivirus (vesivirus and novovirus), feline kobuvirus, feline sakobuvirus A and Lyon IARC polyomaviruses, was detected in 66.1% of the samples.. Co-infections were mainly accounted for by FPV and FCoV and were detected in 24.2% of the samples. The virome composition was further assessed in eight diarrhoeic samples, through the construction of sequencing libraries using a sequence-independent single-primer amplification (SISPA) protocol. The libraries were sequenced on Oxford Nanopore Technologies sequencing platform. A total of 41 contigs (>100 nt) were detected from seven viral families infecting mammals, includedParvoviridae,Caliciviridae,Picornaviridae,Polyomaviridae,Anelloviridae,PapillomaviridaeandParamyxoviridae, revealing a broad variety in the composition of the feline enteric virome.