Abstract

The use of early-stage zebrafish for biomedical research spans early organogenesis to free-swimming larva. A key benefit of this model organism is that repeated assessments spanning several days can be performed of individual larvae within a single experiment, often in conjunction with administered drugs. However, the initiation of feeding, typically at 5 days postfertilization (dpf), can make serial assessments challenging. Therefore, delayed feeding would increase the utility of the model. To ask whether feeding could be delayed without adversely affecting larval growth and development up to 39 dpf, we systematically raised zebrafish and introduced feeding at 5 dpf or delayed initial feeding up to 9 dpf. We assessed survival into the juvenile stage (39 dpf) and anterior-posterior length at this age as proxies for growth and development. Delaying feeding initiation up to 8 dpf did not decrease baseline survival of greater than 90%; survival decreased to 66% only when delayed to 9 dpf. Larval length was no different under any of these conditions. Our findings define 9 dpf as the critical age before which larval zebrafish must be fed when raising to 39 dpf. The option to delay feeding to 8 dpf will broaden experimental applications for the zebrafish larval model.

Highlights

  • Experimentation with zebrafish often requires manipulation and observation after larval hatching and through the first several days when larvae are expected to be free swimming and may be able to initiate feeding

  • All larvae in the group starting feeding at 10 dpf died within 2 days of initial feeding, without significant immediate mortality in the other groups

  • This work demonstrates that delaying the initiation of feeding to 8 dpf does not decrease larval survival and/or growth of zebrafish up to 39 dpf

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Summary

Introduction

Experimentation with zebrafish often requires manipulation and observation after larval hatching and through the first several days when larvae are expected to be free swimming and may be able to initiate feeding. Zebrafish larvae inflate their swim bladder and become free swimming between *96 and 144 h postfertilization (hpf).[1] The age of initial feeding when rearing zebrafish varies but usually coincides with this transition to free swimming, with international zebrafish resource facilities initiating feeding between 5 and 6 days postfertilization (dpf).[2,3] the age range at which zebrafish larvae must initiate feeding for normal survival and maturation into adulthood has not been systematically determined.

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