A Systematic Analysis of Metal and Metalloid Concentrations in Eight Zebrafish Recirculating Water Systems.

Metals and metalloids are integral to biological processes and play key roles in physiology and metabolism. Nonetheless, overexposure to some metals or lack of others can lead to serious health consequences. In this study, eight zebrafish facilities collaborated to generate a multielement analysis of their centralized recirculating water systems. We report a first set of average concentrations for 46 elements detected in zebrafish facilities. Our results help to establish an initial baseline for trouble-shooting purposes, and in general for safe ranges of metal concentrations in recirculating water systems, supporting reproducible scientific research outcomes with zebrafish.

H9c2(2-1)-based sulforhodamine B assay as a possible alternative in vitro platform to investigate effluent and metals toxicity on fish.

To overcome restrictions on the use of fish in toxicity testing, the present study proposes to compare the 50% growth inhibition potential (EC50) of four types of effluents on the rat cardiomyoblast H9c2(2-1) cell line by using the sulforhodamine B (SRB) cell mass colorimetric assay, with the corresponding fish lethal test results. Our objective was to evaluate if H9c2(2-1) cells shows comparable sensitivities, in both relative and absolute terms, to those provided by fish. In parallel, this study also compared the results of the chemical characterization with the legislation in force for environmental protection against effluent release into the receiving environment. Moreover, we tested the H9c2(2-1)-based SRB assays with the metals of concern found in the effluent samples. Both fish and cell assays showed the same toxicity rank for effluents: Metal > Oil > Municipal > Paper, and it should be stressed that the complementarity of using chemical and biological data represents a step forward to guarantee both environmental and human safety, since the chemical characterization showed a different toxicity rank: Metal > Municipal > Oil > Paper. Regarding metal elements, the short-term fish results showed a toxicity rank non-comparable with the rank obtained for cells. Nevertheless, the gathered results reveal the potentiality of the in vitro H9c2(2-1) platform as an alternative for fish lethal testing to assess, in absolute terms, the toxicity of effluents, particularly municipal effluents, and metals.

The Impact of Pseudoloma neurophilia Infection on Body Condition of Zebrafish.

The zebrafish is a widely used animal model in biomedical research. Despite this, pathogens continue to be common in laboratory zebrafish. It is important to determine and describe the pathophysiology of cryptic infections on zebrafish to elucidate the impacts on experimental endpoints. Body condition is a basic measurement used experimentally and in health monitoring of animals. We exposed three wild-type zebrafish strains: AB, WIK, and 5D to Pseudoloma neurophilia. After 8 weeks postexposure, we individually imaged and processed fish for histology. Morphometric analysis was performed on images and an index of body condition was calculated based on the ratio of length/width from the dorsal aspect. Histological sections from each fish were examined to establish sex, severity of infection, and tissue distribution. We observed a significant decrease in body condition in female fish that was not observed in males. In addition, we observed a negative correlation between the total gonadal area of P. neurophilia exposed females and unexposed controls. These results illustrate the sex-specific impacts of a common chronic pathogen on zebrafish health and a commonly used experimental endpoint, further supporting the establishment of rigorous health monitoring programs in laboratory zebrafish colonies that include screening for chronic infectious diseases.

The Impact of Two Different Cold-Extruded Feeds and Feeding Regimens on Zebrafish Survival, Growth and Reproductive Performance.

Zebrafish (Danio rerio) is one of the top model organisms used in biomedical research. Therefore, it is fundamental that zebrafish facilities continuously improve husbandry methods to provide fish with the best physiological and welfare conditions that suit each experimental purpose. Nutrition is a husbandry aspect that needs further optimization, as it greatly affects growth, reproduction, health and behaviour. Here, we have compared the impact of different feeding regimens on zebrafish survival, growth and reproductive performance. Mutant and wild-type zebrafish were raised using several combinations of two cold-extruded processed feeds—Skretting®GemmaMicro and Sparos®Zebrafeed—and one live feed (rotifers). Zebrafeed® outperformed GemmaMicro® in terms of survival rate, and embryo viability was also higher when the spawners were fed with Zebrafeed® either from larval stage or upon sexual maturation. In contrast, GemmaMicro® favoured growth, both in size and weight. The use of rotifers until 60 days post-fertilization improved survival of fish co-fed with GemmaMicro®, while delaying their growth. Zebrafeed® performance was not affected by co-feeding rotifers. Overall, we showed that different nutritional formulas affect physiological parameters, allowing for the establishment of feeding protocols adapted to the objectives of each facility. At the same time, we validated Skretting®GemmaMicro and Sparos®Zebrafeed as two commercially available feeds that are well suited for zebrafish nutrition in a laboratory environment.

Foxj1a is expressed in ependymal precursors, controls central canal position and is activated in new ependymal cells during regeneration in zebrafish.

Zebrafish are able to regenerate the spinal cord and recover motor and sensory functions upon severe injury, through the activation of cells located at the ependymal canal. Here, we show that cells surrounding the ependymal canal in the adult zebrafish spinal cord express Foxj1a. We demonstrate that ependymal cells express Foxj1a from their birth in the embryonic neural tube and that Foxj1a activity is required for the final positioning of the ependymal canal. We also show that in response to spinal cord injury, Foxj1a ependymal cells actively proliferate and contribute to the restoration of the spinal cord structure. Finally, this study reveals that Foxj1a expression in the injured spinal cord is regulated by regulatory elements activated during regeneration. These data establish Foxj1a as a pan-ependymal marker in development, homeostasis and regeneration and may help identify the signals that enable this progenitor population to replace lost cells after spinal cord injury.

Toward an Integrated Zebrafish Health Management Program Supporting Cancer and Neuroscience Research.

Zebrafish is already one of the most used model organisms in biomedical sciences and other research fields. It is therefore becoming increasingly important to assure that zebrafish maintained in laboratory aquaculture conditions are raised and housed under rigorous standards that promote health and welfare to guarantee the required quality and reproducibility of research data. Specifying the programs each facility is adopting would be the first step to achieve this by allowing other facilities to compare, improve, and discuss their protocols and fish performance. We provide in this article a detailed description of an integrated facility health management program, with protocols and readouts, fully designed and aimed at maximizing fish health, welfare, and performance for research.