Enterococcus faecalis is known for its ability to form strong biofilms and its role as an opportunistic pathogen. In this study, we screened and characterized a multidrug-resistant (MDR) and strong biofilm-forming E. faecalis isolate obtained from a shrimp sample to determine its genetic diversity, molecular epidemiology, and underlying factors associated with antimicrobial resistance genes (ARGs) and virulence factor genes (VFGs). The E. faecalis MTR_EFS01 strain was isolated using culturing, staining, and biochemical tests and MALDI-TOF methods. The MDR profile of the strain was determined through the disk diffusion test. The complete genomic sequence of E. faecalis MTR_EFS01 was obtained using the Illumina NextSeq2000 platform. The de novo assembly of the E. faecalis MTR_EFS01 genome revealed a total length of 2,862,301 bp with 80.0 × coverage. This genome comprised 38 contigs, a G + C content of 37.4%, and identified two CRISPR arrays, seven prophages, and 55 RNA genes. The E. faecalis MTR_EFS01 strain was classified as ST862 with a high pathogenicity index of 0.896. The strain harbored eight ARGs conferring resistance to tetracycline, erythromycin, trimethoprim, and MDR efflux pumps. Furthermore, 27 VFGs were identified in this strain, linked to antiphagocytosis, adherence, biofilm formation, enzymes, and immune invasion. Metabolic functional analysis revealed that our strain had 243 subsystems, with the most abundant genes associated with carbohydrate metabolism, amino acids and derivatives, and protein metabolism. The findings in this study underscore the importance of continuous monitoring, research, and collaborative efforts to address the growing threat of MDR and biofilm-forming pathogens in diverse settings.
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