Affordable chironomid housing: proposed modifications of standard OECD substrate for testing of chemicals on aquatic midges

Dimitrija Savić-Zdravković, Aca Đurđević, Zorana Lazarević, Nastasija Manić, Djuradj Milošević


This study was conducted in order to formulate a modification of standard substrate for laboratory bioassays (OECD protocol number 218: ”Sediment water chironomid toxicity test using spiked sediment“), with as few constituents as possible, yet enabling the highest larval survival of model organism: Chironomus tentans (Chironomidae, Diptera). Laboratory experiment consisted of two bioassays: first with substrate mixture of two and more ingredients tested: standard OECD substrate, standard substrate with medical clay, peat + clay, peat + sand, clay + sand; second with every ingredient individually tested: coarse sand, fine sand, peat, clay and no substrate. Highest larval survival was observed in sand + peat (~85%) and coarse sand substrate (~82%), whilst clay + peat (~65%) and peat substrate (~46%) caused the highest larval mortality. No larvae survived in treatment without any substrata, indicating the absolute necessity and importance of substrate presence for larval survival.


Chironomus tentans; bioassay; life traits; OECD guidelines; sediments; substrate; toxicity tests

Full Text:



Brabec, K., Janecek B.F.U., Rossaro B., Spies M., Bitusik P., Syrovatka V. & Schmidt-Kloiber, A. (2017): Dataset "Chironomidae". – An online tool that unifies, standardises and codifies more than 20,000 European freshwater organisms and their ecological preferences. Database version: 7.0 - 10/2016

Bird, G. A. (1997). Deformities in cultured Chironomus tentans larvae and the influence of substrate on growth, survival and mentum wear. Environmental Monitoring and Assessment, 45(3), 273-283.

den Besten, P.J. and Munawar, M., (2016). Ecotoxicological testing of marine and freshwater ecosystems: emerging techniques, trends and strategies. CRC Press.

Davis, J.C., (1977). Standardization and Protocols of Bioassays- Their Role and Significance for Monitoring, Research and Regulatory Usage. In Proceedings of the 3 rd Aquatic Toxicity Workshop, Halifax, Nova Scotia Nov. 2-3, 1976, Environment Canada, Tech. Report.

Dickman, M., & Rygiel, G. (1996). Chironomid larval deformity frequencies, mortality, and diversity in heavy-metal contaminated sediments of a Canadian riverine wetland. Environment International, 22(6), 693-703.

Ferrington, L. C. (2008). Global diversity of non-biting midges (Chironomidae; Insecta-Diptera) in freshwater. Hydrobiologia, 595(1), 447.

Gagliardi, B. S., Pettigrove, V. J., Long, S. M., & Hoffmann, A. A. (2016). A Meta-Analysis Evaluating the Relationship between Aquatic Contaminants and Chironomid Larval Deformities in Laboratory Studies. Environmental science & technology, 50(23), 12903-12911.

Hamilton, A.L. and Saether, O.A., (1971). The occurrence of characteristic deformities in the chironomid larvae of several Canadian lakes. The Canadian Entomologist, 103(3), pp.363-368.

Hund-Rinke, K., Baun, A., Cupi, D., Fernandes, T.F., Handy, R., Kinross, J.H., Navas, J.M., Peijnenburg, W., Schlich, K., Shaw, B.J. and Scott-Fordsmand, J.J., (2016). Regulatory ecotoxicity testing of nanomaterials–proposed modifications of OECD test guidelines based on laboratory experience with silver and titanium dioxide nanoparticles. Nanotoxicology, 10(10), pp.1442-1447. DOI: 10.1080/17435390.2016.1229517

International Organization for Standardization, (2003). Soil quality-guidance on the ecotoxicological characterization of soils and soil materials. ISO.

Langer-Jaesrich, M., Köhler, H. R., & Gerhardt, A. (2010). Can mouth part deformities of Chironomus riparius serve as indicators for water and sediment pollution? A laboratory approach. Journal of soils and sediments, 10(3), 414-422.

Lenat, D. R. (1993). Using mentum deformities of Chironomus larvae to evaluate the effects of toxicity and organic loading in streams. Journal of the North American Benthological Society, 12(3), 265-269.

Meregalli, G., & Ollevier, F. (2001). Exposure of Chironomus riparius larvae to 17α-ethynylestradiol: effects on survival and mouthpart deformities. Science of the total environment, 269(1), 157-161.

Milošević, D., Simić, V., Stojković, M., Čerba, D., Mančev, D., Petrović, A., & Paunović, M. (2013). Spatio-temporal pattern of the Chironomidae community: toward the use of non-biting midges in bioassessment programs. Aquatic ecology, 47(1), 37-55.

OECD 2020, ISSN: 20745761 (online)

No, OECD Test, (2004). 218: Sediment-Water Chironomid Toxicity Using Spiked Sediment. OECD Guidelines for the Testing of Chemicals, Section, 2.

Savić‐Zdravković, D., Djuradj Milošević, Ezgi Uluer, Hatice Duran, Sanja Matić, Snežna Stanić, Janja Vidmar, Janez Ščančar, Domagoj Dikic, and Boris Jovanović. (2020) "A multiparametric approach to cerium oxide nanoparticle toxicity assessment in non‐biting midges." Environmental Toxicology and Chemistry 39, no. 1; 131-140.

Taylor, L. N., & Scroggins, R. P. (2013). Standardization of Ecotoxicological Tests: The Process. Encyclopedia of Aquatic Ecotoxicology, 1073–1080. doi:10.1007/978-94-007-5704-2_9

Wiederholm, T. (1984). Incidence of deformed chironomid larvae (Diptera: Chironomidae) in Swedish lakes. Hydrobiologia, 109(3), 243-249.



  • There are currently no refbacks.