Background: Hookworm species are soil-transmitted helminths that dwell in the small intestine. Albendazole is the first-line drug for hookworm species infection, and it has been given as a treatment for a decade. However, its efficacy has been declining with time. Re-infection has also occurred rapidly after treatment in endemic areas. Regular monitoring of albendazole efficacy and re-infection rates are big challenges in endemic areas. Objective: This study aimed to assess albendazole efficacy, re-infection rate, and factors associated with hookworm species re-infection among schoolchildren in endemic districts of Amhara Region, northwest Ethiopia. Method: A school-based open-level single-arm follow-up study was conducted among 403 schoolchildren from May to December 2022. The study participants were selected by a systematic random sampling technique. Stool samples were processed via Kato-Katz, McMaster, and spontaneous tube sedimentation techniques. All hookworm species -infected schoolchildren were treated with single-dose albendazole (400 mg). The re-infection rates of hookworm species were assessed in the 4th and 6th months of initiation. The data was entered into Epi-data and analyzed using SPSS version 25. The associations of independent variables with the re-infection rate of hookworm species were calculated by bivariate logistic regression. Moreover, all variables with a P-value < 0.20 in the bivariate analysis were run in a multivariate logistic analysis and at 95% CI P-value < 0.05 were considered to be statistically significant. Results: The baseline prevalence of hookworm species was 28.5% (115/403). Among the 115 hookworm species -infected participants, 101 schoolchildren participated in the drug efficacy analysis. The cure rate of albendazole against hookworm species was 85.1% (95% CI 77.3–91.1) by composite reference methods. The egg reduction rate was 93.4% by McMaster and 95.1% by Kato-Katz methods. The re-infection rates of hookworm species in the 4th and 6th months were 23.5% (19/81) and 33.3% (27/81), respectively. Living with > 5 family members (AOR = 4.682, 95% CI = 1.322-16.578, P =.017), poor utilization of latrine (AOR = 5.088, 95% CI = 1.517-17.065, P =.008), infrequent shoes wearing (AOR = 3.099, 95% CI = 1.058-9.083, P =.039), and participating in irrigation (AOR=4.142, 95% CI=1.247-13.753, P=.020) were significantly associated with hookworm species re-infection. Conclusions: Single-dose albendazole was found to be effective against hookworm species infection. The re-infection rates were significant during the 4th and 6th months after treatment. Participating in irrigation, infrequent shoe wear, poor latrine utilization, and livings with large family sizes were identified as risk factors for hookworm species re-infection. Therefore, albendazole should be administered in conjunction with an improved sanitation and hygiene program to control hookworm. Keywords: Hookworm, Albendazole, Re-infection Rates, Cure Rate, Egg Reduction Rate, Ethiopia