EIA is one of the acceptable ways for achieving sustainable development goals and as programming tools, it can define potential environmental impact that appears due to executing development projects, and select logical alternatives for solving them. The objective of this study is environmental impact assessment of Ghameshloo highway on Ghameshloo wildlife refugee condition that is located in Isfehan province. At firs by librarian researches we achieved real recognition of environment of area and the identification of all activities in construction and exploitation stage was implemented by literature reviews of project. Finally ICOLD matrix and checklist method was selected for EIA. The result of research shows that by paying attention to crossing this highway from Ghameshloo wildlife refugee and destroying the environment, the implementing of this plan is rejected. The total Values of plan are +182 and -682. According to being more negative impact (353) in biological environment, this environment incurs a loss more negative impacts from implementing this plan, Social environment with 95 values receives less negative impact