Cross-sectional questionnaire survey was conducted in Arba Minch Zuria and Demba Gofa districts of Gamo and Gofa Zone of the Southern nation nationalities and people’s regional state with the objectives of assessing knowledge of hygienic practice of milk and cheese handlers in both study area. For this a total of 102 woman farmers who are involved in milking, collecting and retailing of milk were included in the study. Data obtained from questionnaire survey were analyzed by descriptive statistics and Chi –square test, using the Statistical package for social science (SPSS Version 19). The participants of this study were woman of different age group with 27(52.9%) and 32(64.7%) of participants in Arba Minch Zuria and Demba Gofa respectively, with mean higher than 36 years old in both study site. The majority of participants were educated up to grade 1-8. A low level of education had an impact on hygienic practice of milking and milk handling. The difference in hygienic handling, training obtained and cheese making practice among the study area were statistically significant (p<0.05). There was also a statistically significant difference in hand washing and utensil cleaning as well as manner of washing between the two study areas (p<0.01). Finally this study revealed that antibiotic usage and sick animal treating practice in was the same in study area (p>0.05) and significant difference in prognosis, Level of skin infection and Selling practice among study participants in study (p<0.05). Good manufacturing; handling practice as well as hygienic milking and cleaning practice should be implemented in both study area

Keywords: Antibiotic Usage; Food Hygiene; Knowledge; Milk and Cheese handling; Woman