We report our recent experience with four cases of endophthalmitis (one male, three females) after 25-gauge pars plana vitrectomy (PPV). One was a case of persistent cystoid macular edema caused by branch retinal vein occlusion, whereas the remaining three were cases of epiretinal membrane. Preoperative antibiotics before the first PPV procedure were not administered in three of the four cases. Endophthalmitis occurred 2-4 days after the first procedure in all cases, for which ceftazidime 2.0 mg/0.1 mL and vancomycin 1.0 mg/0.1 mL were injected into the vitreous cavity. This was followed by emergent 20-gauge PPV and intraocular lens removal using an infusion fluid containing ceftazidime and vancomycin. After the second PPV procedure, progress was good in three cases while retinal detachment occurred in the remaining case one month after surgery; this case required a third PPV procedure. Final best-corrected visual acuity ranged from 20/100 to 20/25 for the four cases. Bacterial cultures were negative after the second PPV procedure in all cases. In conclusion, postoperative endophthalmitis occurred in four of 502 cases (0.80%) that underwent 25-gauge PPV at our hospital. It is important to minimize the incidence of endophthalmitis after 25-gauge PPV.