Antibiotics are indispensable in treating infectious diseases, often administered orally. However, their use can disrupt the delicate balance of gut microbes, affecting the Gut-Brain axis, a bidirectional communication system between the gut microbiota and the nervous system. Such disruptions may lead to neuroinflammation and neurodegeneration. While studies have explored the effects of antibiotics on fear-related behaviors in rodents, the impact on avoidance behaviors, ultrasonic vocalizations (USV) and specific brain regions remains largely unexplored. Our research aims to fill this gap by investigating how oral antibiotic exposure affects avoidance behaviors, USV and brain activity in rodents. By standardizing the acquisition and analysis of USVs and evaluating the effects of antibiotics on avoidance behaviors, we hope to uncover insights into the role of antibiotics in communication and emotional processing. This understanding will help in the targeted interventions addressing anxiety disorders through modulation of the gut-brain axis.