Background: Globally, male involvement has been identified as a priority target area to be strengthened in the prevention of mother to child transmission (PMTCT) of HIV. However, there are limited studies on husband involvement in the prevention of mother to child transmission of HIV in Ethiopia. Therefore, this study aimed to assess male involvement in the prevention of mother to child transmission of HIV and associated factors among males whose wives gave birth in the last six months before the survey in Enebsiesarmider district, Northwest Ethiopia. Methods: A Community-based cross-sectional study was employed to assess male involvement in the prevention of mother to child transmission of human immunodeficiency virus and associated factors in Enebsiesarmider District, Northwest Ethiopia. The study was conducted from February 10-30, 2018. A total of 525 participants were involved in the study. A stratified cluster sampling method was used to recruit study participants. Data were collected using a structured interviewer-administered questionnaire. Data were entered using the epi Data software and exported to SPPS for analysis. Descriptive statistics including mean, a proportion were used to describe study variables. Multivariable logistic regression was employed to describe variables with the outcome variable.Result: Overall male involvement in PMTCT was found to be 26.1% [95%CI, 22.1-29.5]. Respondents who have attended secondary education and above were more likely get involved in PMTCT than who have no formal education [AOR 2.45, 95%CI,, Respondents who have good knowledge on PMTCT [AOR 2.57, 95%CI, 1.58-4.18], good knowledge on ANC [AOR 2.10, 95%CI, 1.28-3.44], low cultural barriers [AOR 2.20, 95%CI, 1.34-3.63] low health system barriers [AOR 2.40, 95%CI, were variables that significantly increase male involvement in PMTCT practices.(Continued on next page)