The purpose of this study is to develop an instrument to assess teachers’ attitudes toward the use of animals. The survey is conducted to explore the relationship between their personal backgrounds and the differences in attitudes toward the use of animals. In addition, this survey aims to examine the correlation between three types of attitudes and three purposes of animal use defined by author. Three types of attitudes include careless use of animals, use of animals under the best animal welfare practice, and never to use animals. The three purposes of animal use involve educational use, scientific purpose, and use for food. The survey containing 36 items is administered to elementary school teachers from Kaohsiung county. 472 valid responses are collected and analyzed using principal component analysis and oblique rotation method subsequently, through which the formal version consisting nine dimensions with 33 items is established. Furthermore, the results indicate that most teachers recognize the use of animals under the best animal welfare practice. It is also found that factors influencing the teachers’ attitudes toward the use of animals include the gender, religious belief, eating habits, experiences of animal breeding, intentions to support social movement and intentions of animal breeding in teaching activities. This study shows positive correlation between the teachers’ attitudes toward the use of animals and their perceptions about the three different purposes of animal use.