Background: The perception and attitude of farmers directly relate to human-animal interactions and these attitudes and perceptions affect farmers’ behavior towards animals’ productivity and animal welfare. To our knowledge, there is no study has been conducted to assess perceptions, attitudes and behavior towards dairy cow production and welfare in Bangladesh. Methodology: A cross-sectional survey was conducted on 270 dairy farmers in Baghabarighat milk shed areas (BMSA) of Sirajganj and Pabna districts in Bangladesh. The study was conducted to assess the perception, attitude, and behavioral practices of animal welfare affecting milk production among animal owners through face-to-face interviews using two sets of questionnaires. The first part represented the socio-demographic characteristics of the participants, and the other parts included the perception, attitude, and behavior of the dairy farmers, which have an impact on milk production. Results: The majority of dairy farmers are engaged in dairy farming but have no formal education or training. Below one fifth knew about animal welfare, and 77.78% of farmers did not agree to provide readily accessible feed and water at all times. Almost 83.33% of dairy farmers thought that reducing unnecessary pain, distress, and mental suffering might decrease the production capability of the dairy cow. It was also believed by all the farmers that good attitudes and behaviors increase milk production. The majority of the farmers stated that negative handling (beat/slap/shouting) during milking decreases milk production, and only a few percentages said that negative handling has no effect on milk production. Almost all of the farmers stated that their economic pressure makes it impossible to maintain the desired level of animal welfare.