Neurobehavioral Effects of Berberine Against Caffeine-Induced Anxiety and Cognitive Impairment in Adult Zebrafish ( Danio rerio ): An Experimental Model

Objectives This study aimed to investigate the effects of two different doses of berberine (BER; 50 and 100 mg L -1 ) on caffeine (CAF; 50 mg L -1 )-induced anxiety-like behaviors and cognitive dysfunction in adult zebrafish ( Danio rerio ). Material and Methods Adult male Danio rerio were divided into 5 groups: control (CTRL), CAF (CAF, 50 mg L -1 ), BER (BER,100 mg L -1 ), (CAF+BER 50), & (CAF+BER100). Fish were exposed to CAF, BER, or their combination for 14 days. Behavioral assays were evaluated as Novel Tank Diving (NTD), Open Field (OF), Light-Dark Preference (LDP), Boldness Assay (BA), Novel Object Recognition (NOR), & T-maze (TM). Results The co-treatment significantly reduced anxiety-like behaviors, boosted exploration as well as boldness, and enhanced cognitive function in a dose-dependent manner, with 100 mg L -1 resulting in higher recovery to CTRL group than BER50. Conclusion BER demonstrated anxiolytic, neuroprotective, and cognitive-enhancing effects, effectively mitigating CAF-induced neurobehavioral impairments. These results highlight its therapeutic potential and support the use of zebrafish as a model in neurobehavioral pharmacology.