Background and Objectives: Due to the increasing development of the poultry industry, having enough information about how to centrally regulate appetite in birds is important. Investigating the mechanisms involved in the central control of appetite in birds is done by ICV injection of biological compounds. In ICV injection in birds, it has been mentioned that only one experiment (one injection) should be done on each bird. This study examines this subject. Materials & Methods: The present study was performed on 24 male Ross 308 broiler chickens. Stereotaxic surgery was performed on all of the animals. After 5 days of recovery, birds every day for a week received ICV injection, and their behavioral changes were examined. Results: No behavioral changes were observed in the first and second days of the experiment. From the third day onwards, the birds show a degree of anxiety, and in the following days these symptoms were more severe and eventually led to the animal death. Repetitive ICV injections may cause inflammation of the brain tissue, and resulting in anxiety symptoms in the animal. Do next ICV injection by intensifying these symptoms, can cause the death of the animal. Conclusion: Repeated ICV injections can cause brain inflammation and anxiety in the animal. Therefore, on every chick just has one experiment (injection) be done.

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