Molecular identification and hydrolase activity profiling of bacteria isolated from septic surface environments using 16S rRNA sequencing.

backgroundHospital septic surfaces represent critical reservoirs for persistent microbial contamination and may contribute to the spread of opportunistic pathogens in healthcare environments. Enzyme-producing bacteria colonizing such surfaces can enhance environmental persistence and resistance to routine sanitation procedures. This study aimed to investigate the diversity, enzymatic potential, and phylogenetic relationships of bacteria isolated from hospital septic surfaces using morphological, biochemical, and molecular approaches.

methodsEnvironmental samples were collected from selected hospital septic surfaces, and bacterial isolates were obtained using standard microbiological techniques. Morphological and biochemical characterization was performed using conventional assays, including catalase, protease, lipase, and amylase activity tests. Genomic DNA was extracted from purified isolates, and the 16 S rRNA gene region was amplified and sequenced. Sequence similarity analysis was performed using BLAST, and phylogenetic relationships were constructed using MEGA11 software based on the Maximum Likelihood method with bootstrap analysis. A total of 28 bacterial isolates were initially obtained, and genomic DNA was extracted from all isolates; however, reliable sequencing data suitable for molecular identification were successfully obtained for 26 isolates.

resultsA total of 26 bacterial isolates were successfully identified from hospital septic environments. Catalase and protease activities were widely observed among isolates, while lipase and amylase activities varied between species. Molecular identification based on 16 S rRNA gene sequencing revealed the predominance of bacterial species belonging to the genera Paenibacillus, Bacillus, and Staphylococcus. Several isolates, including Paenibacillus glucanolyticus and Staphylococcus haemolyticus, showed high sequence similarity (99-100%) with reference strains and formed well-supported phylogenetic clusters. Molecular identification and phylogenetic analysis were performed using 26 successfully sequenced isolates.

conclusionThe presence of diverse enzyme-producing bacterial populations on hospital septic surfaces highlights their potential role in environmental persistence and contamination. These findings emphasize the importance of continuous environmental surveillance and improved sanitation strategies to reduce the risk of microbial dissemination in healthcare settings.