Back ground: Open defecation is a serious sanitation issue globally most developing countries are battling with it. The situation is even worse in the sub-Saharan African countries including Ethiopia, despite various interventions to end open defecation practice. Even if latrine coverage increases with a significant change, Open defecation remains a public health challenge especially in low income countries like Ethiopia including the study area and the prevalence of open defecators and associated factors is not well documented in the study area. Objective: The main objective of the study is to assess the prevalence of open defecation practice and associated factors among households who have latrine in rural communities of Machakle District, Northwest Ethiopia. Methods: a community-based cross-sectional study was conducted from September to October 2019 in Machakle District, Northwest Ethiopia. A total 476 households includes in the study sample and the study population was selected using multistage sampling technique and data was collected using pre-tested questionnaires and observational checklist. Also a total of ten key informant interviews and three focus group discussions were carried out. Result: Open defecation practice among households with latrine 27.8%, 95% CI: (23.8, 32). Educational status of household head (unable to read and write) [(AOR= 5.5, 95% CI: (2.462, 12.36)], occupation of house hold head (being farmer [(AOR=3.25, 95% CI :( 1.7, 6.26)], presence of under five children years in the house [(AOR=3.94, 95% CI:( 2.33,6.67)], latrine cleanness status(being unclean) [(AOR=2.22, 95% CI:( 1.4,3.55) and physical status latrine (latrine need maintenance) [(AOR=2.6, 95% CI:(1.6,4.25)] were significantly associated with open defecation practice. Despite having a private latrine at home or access to a public latrine, people were compelled to practice open defecation due to constraints of habits and hygienic issues in general. Conclusion and recommendation: Latrine construction is not enough to substantially reduce open defecation; indeed, the data concluded that open defecation significantly practiced by households with a latrine. Therefor regular sanitation and hygiene education promotion should be done. Key words: open defecation, households with latrine, Machakle District