The study investigated hygiene issues associated with the lack of cleaning in male masturbators through in vitro microbial culture. The experiment used 24 masturbators simulating human use scenarios, with pig semen serving as a potential microbial culture medium. The results showed that in YPD media, the colony-forming units (CFU) of the no-clean group increased from 11,600 ± 1249 mL⁻¹ at 0 hours to 41,000 ± 8641 mL⁻¹ at 48 hours, representing a 3.53-fold increase. In PDA media, the fungal colony area of the water-cleaned group at 48 hours was 328.01 ± 144.00 mm², while the special cleaning fluid group exhibited only 14.12 ± 3.55 mm². The special cleaning fluid achieved a 97.8% inhibition effect on fungi (PDA media, 48 hours) and a 90.6% cleaning effect on bacteria (BPA media, 48 hours). These findings suggest that the use of GXP special cleaning fluid significantly reduces the risk of microbial growth, ensuring improved hygiene and safety.