Background and Objectives: The emergence of Enterococcus species as a causative agent of Health care Associated Infections, poses therapeutic challenge to clinicians. Enterococci are well equipped with intrinsic resistance to multiple antibiotics. Acquired resistance to commonly used antibiotics such as Penicillin, Aminoglycoside and Vancomycin have made the situation even worse. Detection of High Level Aminoglycoside Resistance (HLAR) in Enterococcus species can predict the loss of synergy between cell wall active antimicrobial agents and Aminoglycosides. Hence, the present study was undertaken to detect the incidence of High level Aminoglycoside Resistant (HLAR) Enterococcus species isolated in a rural hospital and to study their antibiotic susceptibility profile. . Method: HLAR in Enterococcus species was detected by disk diffusion test using High level Streptomycin (HLS - 300 __ampersandsignmu;g) disk and High level Gentamicin (HLG - 120 __ampersandsignmu;g) disk and Agar dilution method as per CLSI guidelines. High Level Gentamicin Resistance (HLGR) was also confirmed by HiMedia Ezy MIC Strip. Antibiotic sensitivity test was done by Kirby Bauer Disc diffusion method as per CLSI guidelines. Result: Out of total 190 Enterococcus species isolated from different clinical samples, High Level Aminoglycoside Resistance was detected in 115 (60.5%) Enterococcus strains. Amongst 115 HLAR Enterococcus strains, 59 (51.4%) were E. faecalis and 56 (48.6%) were E. faecium. Conclusion: We hereby conclude that Enterococcus strains, isolated from different clinical specimens must be screened routinely for HLAR (High Level Aminoglycoside Resistance) by all Clinical Microbiology Laboratories to improve the therapeutic outcome.