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Zebrafish are a preferred vertebrate model for evaluating metabolism during development, and for toxicity studies. However, commercially available intermittent-flow respirometry systems (IFRS) do not provide a suitable zebrafish-scaled swimming tunnel with a low water volume and proper flow velocities. We developed a miniature IFRS (mIFRS) with a 3D-printed, palm-sized zebrafish treadmill for measuring the swimming ability and metabolic rate of a single one- or three-month-old zebrafish with and without toxicity treatment. The 3D-printed zebrafish treadmill consists of discrete components assembled together which enables the provision of a temporary closed circulating water flow. The results showed that three-month-old zebrafish of normal physiological status had higher energetic efficiency and could swim at a higher critical swimming speed (U crit ) of 16.79 cm/s with a lower cost of transport (COT opt ) of 0.11 μmol g -1 m -1 . However, for a single three-month-old zebrafish treated with an antibacterial agent, U crit decreased to 45% of normal zebrafish and the COT opt increased to 0.24 μmol g -1 m -1 , due to the impairment of mitochondria. Our mIFRS provides a low-cost, portable, and readily adaptable tool for studying the swimming performance and energetic metabolism of zebrafish.

A Miniature Intermittent-Flow Respirometry System with a 3D-Printed, Palm-Sized Zebrafish Treadmill for Measuring Rest and Activity Metabolic Rates