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The study was approved by the Institutional Animal Ethics Committee (IAEC) of Periyar University (Approval No. PU-IAEC\/2023\/Zeb.F\/01, dated 30\/08\/2023). Humane endpoints were established prior to experimentation, and fish were monitored daily for signs of stress, abnormal behaviour, or mortality. Animals were maintained under optimal water quality, temperature, and photoperiod conditions to minimize external stressors. No procedures causing unnecessary pain or discomfort were conducted, and the number of animals used was kept to the minimum necessary to achieve statistical validity. 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