Abstract: Objective To understand whether Marmota himalayana is a possible carrier of Enterococcus gallinarum, one of the emerging pathogens for hospital related infections. Methods Selective medium culture, biochemical reaction and 16S rRNA sequencing were conducted for the isolation and identification of isolated strains, K-B test was done to detect the drug susceptibility of isolated strains, and PCR amplification of virulence genes were performed. Methods Three strains of E. gallinarum were isolated from intestinal samples of 79 Marmota himalayana captured in Qinghai-Tibet Plateau in Qinghai province, which were identified by both biochemical tests and 16S rRNA gene sequence analysis. The drug susceptibility test using 15 common antibiotics indicated that 3 isolates were all sensitive to 13 antibiotics, but intermediate to quinupristin, and 1 isolate was intermediate to rifampin. And none of 5 recognized virulence genes in genus of Enterococcus was detected in the 3 isolates, i.e. asa1, esp, hyl, gelE and cylA. Conclusion In contrast to other studies, we isolated 3 antibiotics sensitive and virulence-gene-free E. gallinarum strains from Marmota himalayana for the first time.