Improvement in hand hygiene has been strongly associated with positive outcomes in maternal and child health. Although Bangladesh has a high level of awareness of key hygiene messages, the practice of handwashing, the most effective hygiene behavior, is low. Suchana, a multisectoral nutrition program, aims to increase knowledge and practices around various water, sanitation, and hygiene settings in Sylhet region, the northeast of rural Bangladesh. This study aimed to investigate the changes in indicators related to handwashing knowledge and practices among Suchana beneficiaries in final compared with baseline evaluations. Data were derived from the baseline and final cross-sectional evaluation survey. The following handwashing knowledge and practices were considered: handwashing before preparing food, before eating food and feeding children, before serving food, after defecation, and after touching animals. The descriptive findings indicate that Suchana intervention improved handwashing knowledge and practices in the intervention area compared with the control. The odds of having knowledge of hand washing before preparing food (adjusted odds ratio [aOR]: 1.60; 95% CI: 1.30-1.98), before eating food and feeding children (aOR: 1.68; 95% CI: 1.25-2.25), before serving food (aOR: 1.35; 95% CI: 1.04-1.76), after defecation (aOR: 1.74; 95% CI: 1.25-2.41), and after touching animals (aOR: 1.67; 95% CI: 1.29-2.16) were higher in intervention area than the control area. Similarly, the impact on maternal handwashing practices at final evaluation indicated successful effects of the intervention. These results suggest scaling-up of similar interventions for larger populations living in vulnerable areas of rural Bangladesh.