Tagging Very Small Fish: Two Effective and Low Impact Methods.

Identifying individuals over time and across contexts is essential in many scientific fields. There are a variety of well-established methods for uniquely marking individuals (e.g., visible implant elastomer, barcodes, paint). However, for some species, life history stages, and/or experiments, existing methods are not sufficient. Here, we describe procedures for how two tagging methods-a tattoo ink injection method and a fishing line piercing method - can be used with the youngest, smallest juveniles of the African cichlid fish, Astatotilapia burtoni, which are too small for the methods used with adults. With the tattoo method, we injected tattoo ink into the dorsal muscle. Different colors and injection locations can be used to distinguish among individuals over a period of weeks (up to 4 weeks, average 2.5-3 weeks under our conditions). Because fish this young and small are sensitive to handling and injection, we also include physiological data showing fish recover well from anesthetization and tagging. With the piercing method, very thin fishing line is threaded through the dorsal muscle and tied into a barbell or loop. Unique colors and patterns can be used to distinguish among individuals over a period of months. Because a physical tag might impede normal movement in a very small fish, we also include data from an open field exploration test showing similar behavior between tagged and control (non-tagged) juveniles. We expect these effective and inexpensive methods to be useful for a variety of small species and will facilitate early-life, developmental, and longitudinal research.