The use of Daphnia magna as an experimental animal for such purposes is advantageous in many respects.

Daphnids are small, reaching a size of five mm, so that a great many can be reared in a small space. Further, daphnids are representatives of a class of animals that serve as food for many fish, especially while the fish are young. Fish do not remain in water where their food supply has been depleted. Daphnids would be affected if there was something toxic added to the water, therefore fish would leave and the Daphnia would die. For these reasons Daphnia prove satisfactory for testing on. Daphnia magna is specified to be used in the OECD Guidelines for the Testing of Chemicals, Tests No. 202 "Daphnia sp., Acute Immobilization Test", and Test No. 211 "Daphnia magna Reproduction Test". Test No. 202 is a 48 hour acute toxicity study, where young Daphnia are exposed to varying concentrations of the substance under test and the EC50 determined. Other Daphnia species than Daphnia magna may occasionally be used, but laboratories mostly use Daphnia magna as standard. Test No. 211 is a 21-day chronic toxicity test, at the end of which, the total number of living offspring produced per parent animal alive at the end of the test is assessed, to determine the lowest observed effect concentration of the test substance. (http://en.wikipedia.org/wiki/Daphnia_magna)

Nanoparticle toxicity in Daphnia magna reproduction studies: the importance of test design.

Abstract The increasing use of titanium dioxide nanoparticles (nTiO(2)) inevitably results in their release into the environment, raising concerns about potential adverse effects in wildlife. By following standard test protocols, several studies investigated the ecotoxicity of nTiO(2) among others to Daphnia magna. These studies indicated a large variability - several orders of magnitude - in the response variables. However, other factors, like nanoparticle characteristics and test design, potentially triggering these differences, were largely ignored. Therefore, the present study assessed the chronic ecotoxicity of two nTiO(2) products with varying crystalline structure (A-100; P25) to D. magna. A semi-static and a flow-through exposure scenario were compared, ensuring that both contained environmentally relevant concentrations of dissolved organic carbon. Utilizing the semi-static test design, a concentration as low as 0.06 mg/L A-100 (330 nm) significantly reduced the reproduction of daphnia indicating environmental risk. In contrast, no implication in the number of released offspring was observed during the flow-through experiment with A-100 (140 nm). Likewise, P25 (130 nm) did not adversely affect reproduction irrespective of the test design utilized. Given the present study's results, the particle size, the product composition, i.e. the crystalline structure, and the accumulation of nTiO(2) at the bottom of the test vessel - the latter is relevant for a semi-static test design - may be suggested as factors potentially triggering differences in nTiO(2) toxicity to D. magna. Hence, these factors should be considered to improve environmental risk assessment of nanoparticles. (http://www.ncbi.nlm.nih.gov/pubmed/23202250)

An improved experimental medium for freshwater toxicity studies using Daphnia magna

Abstract Some of the difficulties involved in formulating experimental media for use in toxicity studies are pointed out, with particular reference to heavy metals and their possible chelation by agents in the media. A medium suitable for Daphnia culture with the green alga, Chlorella sp. was chosen from the literature to investigate physiological responses by Daphnia to sublethal cadmium poisoning. Like most synthetic media this included the labile Fe-EDTA complex as a source of soluble iron which is shown to be essential for growth of both Daphnia and Chlorella. Attention is drawn to the almost universal complexing ability of EDTA with metal ions, and to the mechanisms whereby EDTA may be released from the Fe-EDTA complex under experimental conditions. Synthesis of a ferri-gluconate complex as an alternative source of soluble iron is described; this complex being theoretically more iron-specific than Fe-EDTA, whilst providing equally well for Chlorella growth and Daphnia growth and fecundity. Restriction of pH variation in the experimental medium, within its chemically stable range of 6.09.0, is shown to be difficult over 10 days: most buffers with a suitable pKa value (ca. 7.5) being toxic or having some complexing ability. TES buffer (pKa 7.5) and HEPES buffer (pKa 7.55) were used at various concentrations, and 0.001 M HEPES is shown to restrict pH variation to within 0.5 pH units over 10 days without adversely affecting Chlorella growth or Daphnia growth and fecundity. Chemical estimation of the degree of cadmium-complexing by each iron source and each buffer under investigation was not possible since the presence of chelating agents imposes restrictions on the methods available, particularly at low cadmium concentrations. Instead, a bioassay of the reduction in 48 h acute toxicity of cadmium to Daphnia in media containing each potential chelating agent in turn is used. These data indicate that 0.001 M HEPES buffer does not complex cadmium at 0.1 ppm, and that ferri-gluconate is less likely to do so than ferri-EDTA. (http://www.sciencedirect.com/science/article/pii/0043135478900477)

Interlaboratory studies on wastewater toxicity using Daphnia magna Abstract In order to verify the quality of biological measurements, the National Institute of Chemistry in Slovenia is regularly organizing interlaboratory comparisons to estimate the analytical precision of different laboratories using the same test and to determine if statistically significant differences exist between results. Several interlaboratory trials named ILC-Waste Water (ILC-WW) were organized in the last 7 years. Acute toxicity testing with Daphnia magna mobility inhibition assay (ISO 6341) was included in eight successive rounds and about 20 laboratories from Slovenia and a few adjacent countries took part in each study with two samples (T1 and T2) at two different concentration levels. Variation coefficients of the mean 24 h EC50s were determined for each exercise and the evaluation of the interlaboratory variability was analysed. Comparisons were also made of the results and the variability laboratories using organisms taken from laboratory cultures versus those who used daphnias hatched from dormant eggs (Daphtoxkits). (http://link.springer.com/article/10.1007%2Fs00769-009-0519-6)