A Novel Method for Rearing Zebrafish by Using Freshwater Rotifers (Brachionus calyciflorus)

Abstract

The zebrafish (Danio rerio) has become a powerful model organism for studying developmental processes and genetic diseases. However, there remain several problems in previous rearing methods. In this study, we demonstrate a novel method for rearing zebrafish larvae by using a new first food, freshwater rotifers (Brachionus calyciflorus). Feeding experiments indicated that freshwater rotifers are suitable as the first food for newly hatched larval fish. In addition, we revisited and improved a feeding schedule from 5 to 40 days postfertilization. Our feeding method using freshwater rotifers accelerated larval growth. At 49 dpf, one pair out of 10 pairs successfully produced six fertilized eggs. At 56, 63, and 71 dpf, 6 out of the 10 pairs constantly produced normal embryos. Our method will improve the husbandry of the zebrafish.

Contested Paradigm in Raising Zebrafish (Danio rerio)

“…Mean total body length (mm) of zebrafish from present work (Artemia fed) compared to mean body weight of zebrafish achieved in literature through conventional rearing conditions. [4][5][6][7] Continuous light throughout rearing For the first time, we combined constant light with live food availability that secured 24-h feeding/growth of fish for the duration of the first zebrafish generation (43-46 dpf). This is the shortest time from fertilization to the viable egg production reported for zebrafish.…”

“…Current zebrafish rearing procedures are almost uniformly based on diurnal light/dark cycle (14:10 h), provision of live feed at modest density, and high visibility environment. [4][5][6][7][8][9] Consequently, several authors have suggested standardized conditions for larval zebrafish rearing; however, there remains a discrepancy in fish growth and a great variation in fish size at age despite this standardization. For instance, Von Hertel et al 10 reported that the coefficient of variation in zebrafish weight after 4 and 12 weeks of growth was 55% and 60% of the mean, respectively.…”

“…Shortening the generation time (from egg to egg) of zebrafish considerably, compared to earlier rearing results of 90 dpf 45 to 60 dpf, 46 would greatly benefit the research community. By using freshwater rotifers as the initial food, Aoyama et al 5 were able to obtain the first embryos after 46 dpf and consistent reproduction at 50 dpf. Therefore, improving growth at early life stages, when daily growth rate is *15.8%/day at size 5-50 mg and later, at 50-150 mg, when growth rate decreases to 4.7%/day, 8 should significantly shorten generation time.…”

Working with zebrafish at postembryonic stages

“…Typically, rotifers are marine Brachionus plicatilis (Reed Mariculture) that have been acclimated to lower salinities, other varieties of marine and freshwater rotifers can be used as well (Aoyama et al, 2015; Best, Adatto, Cockington, James, & Lawrence, 2010; Lawrence, James, & Mobley, 2015; Lawrence, Sanders, & Henry, 2012). Many rotifer-culturing systems are available, ranging from simple aerated buckets to sophisticated recirculating systems appropriate for a large, high-throughput fish facility.…”

Diving into the world of alcohol teratogenesis: a review of zebrafish models of fetal alcohol spectrum disorder

“…George Streisinger was drawn to the zebrafish due to several characteristics (Streisinger et al 1981). First, zebrafish have a short generation time, with recent advances in rearing resulting in times of approximately 60 days (Aoyama et al 2015;Mahabir et al 2013). Second, zebrafish are highly fecund in the laboratory, with a single female being able to lay hundreds of eggs once per week.…”