Improving Production of Zebra Fish Embryos in the Lab

Robert Ohene Adu, Jens Peter Thomsen
DOI: 10.4236/jep.2011.210157                

Abstract

The utilization of fish embryos in toxicity testing of hazardous chemicals has recently been adopted in order to satisfy stricter rules and regulations related to using adult animals in toxicity testing. This paper presents optimising steps towards improving zebra fish embryo production in the laboratory. Culture conditions were maintained in the aquaria as stipulated in the OECD draft proposal for a new guideline on fish embryo tests. Furthermore, a sequence of steps were adopted and followed to improve upon previous work done in the lab in 2006. About 200 eggs were produced in one spawn trap within an hour of onset of light, an improvement over the 50 - 60 eggs produced in the previous work. This result demonstrates that with the right culture conditions and proper optimisation of procedure the required number of embryos needed for toxicity testing can be obtained.

Keywords

Hazardous Chemicals, Zebra Fish Embryo, Toxicity Testing, Spawn Trap, Culture Conditions

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	S. Scholz, S. Fischer, U. Guundel, E. Kuuster, T. Luckenbach and D. Voelker, “The Zebrafish Embryo Model in Environmental Risk Assessment-Applications beyond Acute Toxicity Testing,” Environmental Science and Pollution Research, Vol. 15, No. 5, 2008, pp. 394-404.
[2]	G. Schüürmann, K. Schirmer, E. Küster, B. Hansjürgens and W. K?ck, “Using REACH as an Opportunity to Find Alternatives to Animal Experiments: A Need for Intelligent Test Strategies Instead of Blindly Testing Everything,” 2005. http://www.ufz.de/index.php?en=6556
[3]	V. J. Brown, “Reaching for Chemical Safety,” 2003. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1241736/pdf/ehp0111-a00766.pdf
[4]	T. Petry, R. Knowles and R. Meads, “An Analysis of the Proposed REACH Regulations,” 2006. http://toxminds.org/pdf_publications/TP-REACH-RTP-2006.pdf
[5]	R. Combes, M. Baratt and M. Balls, “An Overall Strategy for the Testing of Human Hazard and Risk Assessment under the EU REACH System,” Alternatives to Laboratory Animals, Vol. 31, No. 1, 2003, pp. 7-19.
[6]	W. Lilienblum, W. Dekant, H. Foth, T. Gebel, J. G. Hengstler, R. Kahl, P. J. Kramer, H. Schweinfurth and K. M. Wollin, “Alternative Methods to Safety Studies in Expe- rimental Animals: Role in the Risk Assessment of Che- micals under the New European Chemicals Legislation (REACH),” 2008. http://www.ncbi.nlm.nih.gov/pubmed/18322675
[7]	E. Lammer, G. J. Carr K. Wendler; J. M. Rawlings, S. E. Belanger and Th. Braunbeck, “Is the Fish Embryo Toxicity Test (FET) with the Zebra Fish (Danio rerio) a Potential Alternative for the Fish Toxicity Test?” Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, Vol. 149, No. 2, 2009, pp. 196-209. doi:10.1016/j.cbpc.2008.11.006
[8]	T. Braunbeck and E. Lammer, “Background Paper on Fish Embryo Toxicity Assays. Prepared for German Federal En- vironment Agency,” UBA Contract No. 203 85 422, 2006.
[9]	K. Schirmer, K. Tanneberger, N. I. Kramer, D. V?lker, S. Scholz, C. Hafner, E. J. Lee, N. C. Bols and J. L. M. Her- mens, “Developing a List of Reference Chemicals for Test- ing Alternatives to Whole Fish Toxicity Tests,” 2008. http://www.wlu.ca/documents/32518/Schirmer_et_al_aqua_tox08.pdf
[10]	S. M. Hassan and E. A. Moussa, “Effects of Quillaja Sa- ponin (Quillaja saponaria) on Early Embryonic Zebrafish (Danio rerio) Development,” International Journal of To- xicology, Vol. 27, No. 3, 2008, pp. 273-278. doi:10.1080/10915810802152129
[11]	S. B. Frandsen, “Chestnuts Cause Fish Kills,” 2010. http://www.kolding.dk/data/0067955.asp?sid=13552&uid=24949
[12]	M. Wirtz, “Toxicity and Possible Biodegradation of Ground- water from a Former Chemical Dump Site (Kaaergard Plantage),” University of Applied Sciences Oldenburg/Ostfries- land/Wilhelmshaven; Aalborg University, Esbjerg, 2006.
[13]	OECD, “OECD Guideline for the Testing of Chemicals,” Draft Proposal for a New Guideline: Fish Embryo Toxicity (FET) Test, 2006.
[14]	F. Lahnsteiner, “The Sensitivity and Reproducibility of the Zebrafish (Danio rerio) Embryo Test for the Screening of Waste Water Quality and for Testing the Toxicity of Chemicals,” ATLA. Alternatives to Laboratory Animals, Vol. 36, No. 3, 2008, pp. 299-311.