Enterococcus faecalisandEnterococcus faeciumhave become two of the most important agents of nosocomial diseases due to their constantly growing resistance. Enterococcal infections are associated with biofilms, which are intrinsically sensitive to antimicrobials. The main goal of this study was to compare and relate their capacity to form biofilm and their antimicrobial sensitivity, as well as their virulence factors and their implicated genes, of strains isolated from patients with urinary tract infection (UTI) in a rural hospital in Uganda and a secondary hospital in Spain. A prospective study was conducted with 104 strains ofE. faecalisandE. faeciumisolated from patients with suspected UTI and who presented leukocyturia at the Saint Joseph Kitgum hospital (Uganda) and at the Hospital Universitario Principe de Asturias (Spain). All microorganisms were identified in Spain by MALDI-TOF mass spectrometry. Antimicrobial susceptibility studies were carried out using the Vitek®2 system (Biomériux, France). The biofilm formation capacity was studied by photospectrometry. Phenotypic and genotypic virulence factors were studied in all cases by PCR or expression techniques. In Uganda, we found a higher incidence ofE. faecium(65.3%,n= 32), contrary to the situation found in Spain where most of the bacteria found belonged toE. faecalis(92.7%,n= 51). AllE. faecalisstrains were found to have very low levels of resistance to ampicillin, imipenem, and nitrofurantoin. However,E. faeciumexhibited more than 25% resistance to these antibiotics. Although theespgene has been shown in the results obtained to be an important initial agent in biofilm formation, we have also demonstrated in this study the intervention of other genes whenespis not present, such as theace1gene. No statistically significant relationships were found between the presence ofaggandgelEgenes and increased biofilm formation. The significant difference between the incidence ofE. faecalisandE. faeciumand biofilm formation, between samples from Spain and Uganda, shows us very different profiles between countries.