Environmental Science and Pollution Research

, Volume 25, Issue 11, pp 10730–10739 | Cite as

Ecotoxicity of the nonsteroidal ecdysone mimic RH-5849 to Daphnia magna

  • Jinlin Jiang
  • Zhengjun Shan
  • Xiaorong Wang
  • Yuxuan Zhu
  • Junying Zhou
Research Article


The nonsteroidal ecdysone mimic 1,2-dibenzoyl-1-tert-butylhydrazine (RH-5849), a novel insect growth regulator, is mainly registered for use in rice fields. So far, its toxicity and ecological risks towards aquatic cladoceran invertebrates remain unclear. In this study, RH-5849 was evaluated for its acute and 21-day chronic toxicity towards Daphnia magna. The viability, morphology, growth, and reproduction of D. magna were observed to establish a concentration-toxicity relationship associated with the RH-5849 exposure. In addition, the relationship between the changes of physiological and biochemical indices and the chronic indices was analyzed in order to find potential early warning indicators in D. magna to the chronic risk of RH-5849 exposure. The results showed that the 48-h EC50 of acute immobilization and EC50 of 21-day survival of RH-5849 on D. magna were 45.3 and 1.34 mg/L, respectively. Chronic exposure to RH-5849 mainly affected the reproductive parameters of D. magna and the no observed effect concentration (NOEC) and the EC50 were 0.050 and 0.5423 mg/L, respectively. The number of offspring per female reduced significantly after 21-day exposure to 0.10 mg/L of RH-5849. The morphological changes, manifested in head width and body length, the length of the helmet or apical spine, and the curvature and transparency of the body, were observed in RH-5849-treated groups. Moreover, it was found that the alkaline phosphatase activity in D. magna after 5–7-day exposure was positively correlated with the number of offspring per female after 21 days. These results indicate the potential risk of RH-5849 towards aquatic crustaceans should be taken into consideration when applied to rice fields.


RH-5849 Daphnia magna Chronic toxicity Growth Reproduction 


Funding information

This research was financially supported by the National Natural Science Foundation of China (Grant number 21407056).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Supplementary material

11356_2018_1275_Fig8_ESM.jpg (9 kb)
Figure S1

Mean survival time of Daphnia magna under different RH-5849 concentrations during the 21-day experiment. Data are denoted as mean ± standard deviation. No asterisk indicates there was no statistically significant difference in the treatment relative to control. (JPEG 8 kb)

11356_2018_1275_MOESM1_ESM.tif (610 kb)
High resolution image (TIFF 609 kb)


  1. Bagheri F, Talebi K, Hosseininaveh V (2010) Cellular energy allocation of pistachio green stink bug, Brachynema germari Kol. (Hemiptera.: Pentatomidae) in relation to juvenoid pyriproxyfen[J]. Afr J Biotechnol 9:5746–5753Google Scholar
  2. Bengochea P, Christiaens O, Amor F, Viñuela E, Rougé P, Medina P, Smagghe G (2013) Insect growth regulators as potential insecticides to control olive fruit fly (Bactrocera oleae Rossi): insect toxicity bioassays and molecular docking approach. Pest Manag Sci 69:27–34CrossRefGoogle Scholar
  3. Biesinger KE, Christensen GM (1972) Effects of various metals on survival, growth, reproduction, and metabolism of Daphnia magna. J Fish Res Board Can 29:1691–1700CrossRefGoogle Scholar
  4. Bogdan KG, Gilbert JJ (1984) Body size and food size in freshwater zooplankton. Proc Natl Acad Sci 81:6427–6431CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  6. Chèvre N, Becker-van Slooten K, Tarradellas J, Brazzale AR, Behra R, Guettinger H (2002) Effects of dinoseb on the life cycle of Daphnia magna: modeling survival time and a proposal for an alternative to the no-observed-effect concentration. Environ Toxicol Chem 21:828–833CrossRefGoogle Scholar
  7. Chèvre N, Brazzale AR, Becker-van Slooten K, Behra R, Tarradellas J, Guettinger H (2005) Modeling the concentration-response function of the herbicide dinoseb on Daphnia magna (survival time, reproduction) and Pseudokirchneriella subcapitata (growth rate). Ecotoxicol Environ Saf 62:17–25CrossRefGoogle Scholar
  8. Clare AS, Rittschof D, Costlow JD (1992) Effects of the nonsteroidal ecdysone mimic RH 5849 on larval crustaceans. J Exp Zool 262:436–440CrossRefGoogle Scholar
  9. Coen WMD, Janssen CR (1998) The use of biomarkers in Daphnia magna toxicity testing. Hydrobiologia 367:199–209CrossRefGoogle Scholar
  10. Darvas B, Polgar L, Tag El-din MH, Eross K, Wing KD (1992) Developmental disturbances in different insect orders caused by an ecdysteroid agonist, RH5849. J Econ Entomol 85:2107–2112CrossRefGoogle Scholar
  11. Dhadialla TS, Carlson GR, Le DP (1998) New insecticides with ecdysteroidal and juvenile hormone activity[J]. Annu Rev Entomol 43:545–569CrossRefGoogle Scholar
  12. Du YZ, Liu AX (2002) Research advance on nonsteroid ecdysone agonist. Pesticide 4: 7–11. (In Chinese)Google Scholar
  13. Elendt BP, Bias WR (1990) Trace nutrient deficiency in Daphnia magna cultured in standard medium for toxicity testing.Effects of the optimization of culture conditions on life history parameters of D. magna. Water Res 24:1157–1167CrossRefGoogle Scholar
  14. Jiang JL, Shi Y, Shan ZJ, Yang LY, Wang XR, Shi LL (2012) Bioaccumulation, oxidative stress and HSP70 expression in Cyprinus carpio L. exposed to microcystin-LR under laboratory conditions. Comp Biochem Physiol, Part C 155:483–490Google Scholar
  15. Ke ZM, Huang LM (2009) Advance in the study on the anti-predator phenotypic plasticity of Daphnia. J Lake Sci 21:758–767CrossRefGoogle Scholar
  16. Miller DM, Aust SD (1989) Studies of ascorbate-dependent, iron catalyzed lipid peroxidation. Arch Biochem Biophys 271:113–119CrossRefGoogle Scholar
  17. MOA PRC (2014) Test guidelines on environmental safety assessment for chemical pesticides―part 13: Daphnia sp. acute immobilisation test. GB/T 31270.13–2014.Google Scholar
  18. Monthean C, Potter DA (1992) Effects of RH5849, a novel insect growth regulator, in Japanese beetle (Coleoptera: Scarabaeidae) and fall armyworm (Lepidoptera: Noctuidae) in turfgrass. J Econ Entomol 85:507–513CrossRefGoogle Scholar
  19. OECD (1998) Test guideline no. 211. Test guidelines for testing of chemicals: Daphnia magna reproduction test. Paris: OECD Publishing.Google Scholar
  20. OECD (2004) Test guideline no. 202. Test guidelines for testing of chemicals: Daphnia sp. acute immobilisation test. Paris: OECD Publishing.Google Scholar
  21. OECD (2008) Test guideline no. 211. Effects on biotic systems test: Daphnia magna reproduction test. Paris: OECD Publishing.Google Scholar
  22. Ou J, Zhu X, Wang L, Xu C, Liu F, Ren L, Xu X, Wang Y, Rui C, Liu S (2012) Synthesis and bioactivity study of 2-acylamino-substituted N'-benzylbenzohydrazide derivatives. Agric Food Chem 60:10942–10951CrossRefGoogle Scholar
  23. Riddiford LM, Cherbas P, Truman JW (2000) Ecdysone receptors and their biological actions. Vitam Horm 60:1–73CrossRefGoogle Scholar
  24. Rui CH, Liu J, Ren L (2012) Toxicological mechanisms and resistance of insect growth regulators. J Biosafety 21: 177–183. (In Chinese)Google Scholar
  25. Salgado VL (1992) The neuro-toxic insecticidal mechanism of the nonst eroidal ecdysone agonist RH-5849: K+ channel block in nerve and muscle. Pestic Biochem Physiol 43:1–13CrossRefGoogle Scholar
  26. Stauber JL, Florence TM (1989) The effect of culture medium on metal toxicity to the marine diatom Nitzschia closterium and the freshwater green alga Chlorella pyrenoidosa. Water Res 23:907–911CrossRefGoogle Scholar
  27. Song MY, Stark JD, Brown JJ (1997) Comparative toxicity of four insecticides, including imidacloprid and tebufenozide, to four aquatic arthropods. Environ Toxicol Chem 16:2494–2500CrossRefGoogle Scholar
  28. Sumiya E, Ogino Y, Toyota K, Miyakawa H, Miyagawa S, Iguchi T (2016) Neverland regulates embryonic moltings through the regulation of ecdysteroid synthesis in the water flea Daphnia magna, and may thus act as a target for chemical disruption of molting. J Appl Toxicol 36:1476–1485CrossRefGoogle Scholar
  29. Sumiya E, Ogino Y, Miyakawa H, Hiruta C, Toyota K, Miyagawa S, Iguchi T (2014) Roles of ecdysteroids for progression of reproductive cycle in the fresh water crustacean Daphnia magna. Front Zool 11:1–12CrossRefGoogle Scholar
  30. Tran HT, Askari HB, Shaaban S, Price L, Palli SR, Dhadialla TS, Carlson GR, Butt TR (2001) Reconstruction of ligand-dependent transactivation of Choristoneura fumiferana ecdysone receptor in yeast. Mol Endocrinol 15:1140–1153CrossRefGoogle Scholar
  31. Trisyono A, Chippendale GM (1997) Effect of the nonst ertoidal ecdysone agonists, methoxyfenozide and tebuf enozide, on the European corn borer (Lepidoptera: Pyralidae). J Econ Entomol 90:1486–1492CrossRefGoogle Scholar
  32. Wagner ND, Frost PC (2012) Responses of alkaline phosphatase activity in Daphnia to poor nutrition. Oecologia 170:1CrossRefGoogle Scholar
  33. Ward CJ, Beattie KA, Lee EY, Codd GA (1997) Colorimetric protein phosphatase inhibition assay of laboratory strains and natural blooms of cyanobacteria: comparisons with high-performance liquid chromatographic analysis for microcystins. FEMS Microbiol Lett 153:465–473CrossRefGoogle Scholar
  34. Wiegand C, Pflugmacher S (2005) Ecotoxicological effects of selected cyanobacterial secondary metabolites a short review. Toxicol Appl Pharmacol 203:201–218CrossRefGoogle Scholar
  35. Wing KD, Slawecki RA, Carlson GR (1988) RH 5849, a nonsteroidal ecdysone agonist: effects on larval Lepidoptera. Science 241:470–472CrossRefGoogle Scholar
  36. Xiang FH, Yang W, Yang Z, Chen YF (2011) Concentration–response function of nitrite on survival, molting, and reproduction of Daphnia similoides. J Freshw Ecol 26:33–41CrossRefGoogle Scholar
  37. Xu H, Li Z (2007) Residue and degradation of RH-5849 in rice and soil. J Agro-Environ Sci 26:1759–1763Google Scholar
  38. Yokota K, Sterner RW (2011) Trade-offs limiting the evolution of coloniality: ecological displacement rates used to measure small costs. Proc Royal Soc B Biol Sci 278:458–463CrossRefGoogle Scholar
  39. Zhao XF, Liu GX, Hu ZY (2006) Physico-chemical properties of alkaline phosphatases released by a planktonic crustacean, Daphnia magna (cladocera). Crustaceana 79:677–689CrossRefGoogle Scholar
  40. Zhong Y, Zang Y, Luo Y, Kong ZM, Shen JP (1999) Toxicological study of two new pesticides on earthworms. Agro-environ Protection 3: 102–105. (In Chinese)Google Scholar
  41. Zhu YX, Jiang JL, Shan ZJ, Bu YQ, Xu WL, Cheng Y (2014) Acute and chronic toxicity of RH-5849 to Daphnia magna. J Agro-Environ Sci 33: 2309–2314. (In Chinese)Google Scholar
  42. Ziarek JJ, Nihongi A, Nagai T, Uttieri M, Rudi SJ (2011) Seasonal adaptations of Daphnia pulicaria swimming behaviour: the effect of water temperature. Hydrobiologia 661:317–327CrossRefGoogle Scholar
  43. Zou E, Fingerman M (1997) Effects of estrogenic xenobiotics on molting of the water flea, Daphnia magna. Ecotoxicol Environ Saf 38:281–285CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Jinlin Jiang
    • 1
  • Zhengjun Shan
    • 1
  • Xiaorong Wang
    • 2
  • Yuxuan Zhu
    • 1
  • Junying Zhou
    • 1
  1. 1.Nanjing Institute of Environmental Sciences/Key Laboratory of Pesticide Environmental Assessment and Pollution ControlMinistry of Environmental ProtectionNanjingPeople’s Republic of China
  2. 2.School of the EnvironmentNanjing UniversityNanjingPeople’s Republic of China

Personalised recommendations