T errestrial hot springs offer unique habitats for various bacteriophages, revealing potential reservoirs and a library for antibiotic-resistant and biofilm-removing bacteriophages. This study aimed to identify biofilm-removing bacteriophages as a potential method to eradicate antibiotic-resistant bacteria. Samples were collected from the Mae Chan Fault geothermal sites in Chiang Rai province, north of Thailand. Our bacteriophages, HSPT1 and HSPT2, effectively infect Pseudomonas aeruginosa TISTR1287, as measured by their plaque sizes (3.23 ± 0.26 mm) and (1.14 ± 0.18 mm), respectively. Thermostability analysis reveals that the optimal temperature for bacteriophage HSPT1 is between 20 and 60 oC, while for bacteriophage HSPT2, it is between 20 and 40 oC. Biofilm biomass was significantly reduced by the bacteriophages HSPT1 and HSPT2 treatments (1.42 ± 0.81 and 2.51 ± 0.71, respectively). The study reveals that hot spring bacteriophages effectively remove bacterial biofilms, proving their potential as a new treatment option for antibiotic-resistant bacteria in clinical and tourism settings. These bacteriophages, derived from hot springs, have effectively reduced biofilm-forming microorganisms and multidrug-resistant pathogenic bacteria.