The Complete and Updated "Rotifer Polyculture Method" for Rearing First Feeding Zebrafish

Abstract

The zebrafish (Danio rerio) is a model organism of increasing importance in many fields of science. One of the most demanding technical aspects of culture of this species in the laboratory is rearing first-feeding larvae to the juvenile stage with high rates of growth and survival. The central management challenge of this developmental period revolves around delivering highly nutritious feed items to the fish on a nearly continuous basis without compromising water quality. Because larval zebrafish are well-adapted to feed on small zooplankton in the water column, live prey items such as brachionid rotifers, Artemia, and Paramecium are widely recognized as the feeds of choice, at least until the fish reach the juvenile stage and are able to efficiently feed on processed diets. This protocol describes a method whereby newly hatched zebrafish larvae are cultured together with live saltwater rotifers (Brachionus plicatilis) in the same system. This polyculture approach provides fish with an "on-demand", nutrient-rich live food source without producing chemical waste at levels that would otherwise limit performance. Importantly, because the system harnesses both the natural high productivity of the rotifers and the behavioral preferences of the fish, the labor involved with maintenance is low. The following protocol details an updated, step-by-step procedure that incorporates rotifer production (scalable to any desired level) for use in a polyculture of zebrafish larvae and rotifers that promotes maximal performance during the first 5 days of exogenous feeding. Video LinkThe video component of this article can be found at

In vivo imaging of calcium dynamics in zebrafish hepatocytes

“…Subsequently, zebrafish larvae display a feeding to fasting transition and enter a starvation state. To test if calcium oscillations in the liver depend on nutrient availability, we fed larvae starting at 5 dpf, which is the standard feeding schedule for zebrafish rearing (Lawrence et al, 2016). At 7dpf, the livers of fed animals displayed synchronized calcium oscillations (Fig.…”

zPACT: Tissue Clearing and Immunohistochemistry on Juvenile Zebrafish Brain

“…Fish are reared for 6 weeks under controlled conditions (photoperiod = 14 h/10 h light/dark; temperature = 26.5 ± 1 °C; pH = 7.8 ± 0.1; conductivity = 240 ± 30 µS/cm, NH4 + = 0 mg/L, NO2 -= 0 mg/L, NO3 -< 50 mg/L), in groups of 50 individuals per 1.4 L tank. They are fed on rotifers (Brachionus plicatilis; > 1,000/fish/day; one dose at noon) for the first two weeks of feeding, and then with brine shrimps (Artemia nauplii; ~250/fish/day) and dry food (Skretting, Gemma Micro; to apparent satiation, two doses, morning, afternoon) (Lawrence and Mason, 2012;Lawrence et al, 2016). All procedures are performed in accordance with European Union Directive 2010/63/EU.…”

The Vital Relationship Between Nutrition and Health in Zebrafish

“…12 Live diets are clearly superior to processed feeds during early larval stages of development. [37][38][39][40] Furthermore, there has been a recent movement, especially in Europe, to provide live prey items throughout the entire life cycle as a form of environmental enrichment. 41,42 Therefore, if live feeds are retained in the diet, the recommendation would be to subject these items to routine screening to reduce the risk of contamination by pathogens.…”