Journal of Applied Phycology

, Volume 31, Issue 1, pp 41–48 | Cite as

The natural triterpenoid toosendanin as a potential control agent of the ciliate Stylonychia mytilus in microalgal cultures

  • Ran Xu
  • Litao Zhang
  • Jianguo LiuEmail author


With the aim of identifying an effective and safe technique for reducing contamination by the ciliate, Stylonychia mytilus, in microalgal cultivation, the toxic effects of the natural triterpenoid toosendanin and ammonium bicarbonate on S. mytilus were studied. Toxicity tests showed that toosendanin and ammonium bicarbonate were highly toxic to S. mytilus, with 24 h LC50 values of 6.4 μg L−1 and 0.8 g L−1, respectively. The population density of S. mytilus decreased significantly when exposed to ≥ 2 μg L−1 toosendanin or ≥ 0.4 g L−1 ammonium bicarbonate. In addition, the S. mytilus control effects of toosendanin and ammonium bicarbonate and their safety in Chlorella pyrenoidosa were evaluated. It was found that ≤ 14 μg L−1 toosendanin had no obvious influence on photosynthesis and growth of C. pyrenoidosa and even increased the final cell density, with the highest being 12.3% over that of untreated cultures. Ammonium bicarbonate is the most widely used optimization technique for controlling contamination, but it has limited ability to reduce S. mytilus. Furthermore, ≥ 0.8 g L−1 ammonium bicarbonate inhibited photosynthesis and growth of C. pyrenoidosa, causing a 5.1% reduction in cell density or even a complete crop failure. Based on its high toxicity to S. mytilus and its relative safety to C. pyrenoidosa, together with its low commercial price and ecological acceptability, toosendanin is considered to be a good potential botanical pesticide for controlling S. mytilus contamination in microalgal mass cultivation.


Toosendanin Ammonium bicarbonate Stylonychia mytilus Contamination control Microalgal cultivation 



This work was supported by National Key Research and Development Program—China (2016YFB0601004) and Hainan Aoji Biological Technology Co., Ltd. We thank Dr. John van der Meer (Pan-American Marine Biotechnology Association) for his assistance with proofreading.


  1. Ammermann D (2008) Cell size and dorsal cilia: are they useful features for the identification of Stylonychia mytilus (Ciliophora, Spirotrichea) and its subpopulations? Eur J Protistol 44:263–268CrossRefGoogle Scholar
  2. Acosta F, Zamor RM, Najar FZ, Roe BA, Hambright KD (2015) Dynamics of an experimental microbial invasion. Proc Natl Acad Sci U S A 112:11594–11599CrossRefGoogle Scholar
  3. Araújo CV, Tornero V, Lubián LM et al (2010) Ring test for whole-sediment toxicity assay with a benthic marine diatom. Sci Total Environ 408:822–828Google Scholar
  4. Atkins EL, Greywood EA, Macdonald RL (1975) Toxicity of pesticides and other agricultural chemicals to honey bees. University of California, Division of Agricultural Sciences, Leaflet 2287:38–42Google Scholar
  5. Borowitzka MA (2013) High-value products from microalgae—their development and commercialisation. J Appl Phycol 25:743–756CrossRefGoogle Scholar
  6. Borowitzka MA, Moheimani NR (2013) Open pond culture systems. In: Borowitzka MA, Moheimani NR (eds) Algae for biofuels and energy. Springer, Dordrecht, pp 133–152CrossRefGoogle Scholar
  7. Chalifour A, Spear PA, Boily MH, Deblois C, Giroux I, Dassylva N (2009) Assessment of toxic effects of pesticide extracts on different green algal species by using chlorophyll a fluorescence. Toxicol Environ Chem 91:1315–1329CrossRefGoogle Scholar
  8. Chen W, Isman MB, Chiu SF (2010) Antifeedant and growth inhibitory effects of the limonoid toosendanin and Melia toosendan extracts on the variegated cutworm, Peridromasaucia (Lep., Noctuidae). J Appl Entomol 119:367–370CrossRefGoogle Scholar
  9. Cosgrove J, Borowitzka MA (2011) Chlorophyll fluorescence terminology: an introduction. In: Suggett DJ, Prásil O, Borowitzka MA (eds) Chlorophyll a fluorescence in aquatic sciences: methods and applications. Springer, Dordrecht, pp 1–17Google Scholar
  10. Huang Y, Liu JG, Yu WH, Gao GZ (2014a) Treatment potential of a synergistic botanical pesticide combination for rotifer extermination during outdoor mass cultivation of S. platensis. Algal Res 6:139–144CrossRefGoogle Scholar
  11. Huang Y, Liu JG, Li L, Pang T, Zhang LT (2014b) Efficacy of binary combinations of botanical pesticides for rotifer elimination in microalgal cultivation. Bioresour Technol 154:67–73CrossRefGoogle Scholar
  12. Huang Y, Liu JG, Li L, Pong T (2017) Growth inhibitory and antifeedant effects of sublethal concentrations of toosendanin on the rotifer Brachionus plicatilis. Biomass Bioenergy 99:31–37CrossRefGoogle Scholar
  13. Ilavarasi A, Mubarakali D, Praveenkumar R, Baldev E, Thajuddin N (2011) Optimization of various growth media to freshwater microalgae for biomass production. Biotechnology 10:540–545CrossRefGoogle Scholar
  14. Junghans M, Backhaus T, Faust M, Scholze M, Grimme LH (2003) Predictability of combined effects of eight chloroacetanilide herbicides on algal reproduction. Pest Manag Sci 59:1101–1110CrossRefGoogle Scholar
  15. Lam MK, Lee KT (2012) Microalgae biofuels: a critical review of issues, problems and the way forward. Biotechnol Adv 30:673–690CrossRefGoogle Scholar
  16. Lincoln EP, Hall TW, Koopman B (1983) Zooplankton control in algal cultures. Aquaculture 32:331–337CrossRefGoogle Scholar
  17. Ma ZQ, Monika GN, Xing Z, Mark RB (2013) Effects of the botanical insecticide, toosendanin, on blood digestion and egg production by female Aedes aegypti (Diptera: Culicidae): topical application and ingestion. J Med Entomol 50:112–121CrossRefGoogle Scholar
  18. Ong ES, Ong CN (2007) Qualitative and quantitative analysis of toosendanin in Melia toosendan Sieb. Et Zucc (Meliaceae) with liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 21:589–598CrossRefGoogle Scholar
  19. Pang T, Liu JG, Liu Q, Zhang LT, Lin W (2012) Impacts of glyphosate on photosynthetic behaviors in Kappaphycus alvarezii and Neosiphonia savatieri detected by JIP-test. J Appl Phycol 24:467–473CrossRefGoogle Scholar
  20. Post FJ, Borowitzka LJ, Borowitzka MA, Mackay B, Moulton T (1983) The protozoa of a western Australian hypersaline lagoon. Hydrobiologia 105:95–113CrossRefGoogle Scholar
  21. Shi XB, Frankel J (1990) Morphology and development of mirror image doublets of Stylonychia mytilus. J Protozool 37:1–13CrossRefGoogle Scholar
  22. Shi YL, Li MF (2007) Biological effects of toosendanin, a triterpenoid extracted from Chinese traditional medicine. Prog Neurobiol 82:1–10CrossRefGoogle Scholar
  23. Santosh K, Daizy B, Pablo QA, Mann KS, Antonietta LT (2016) Fine-tune investigations on three stylonychid (Ciliophora, Hypotricha) ciliates. Eur J Protistol 56:200–218CrossRefGoogle Scholar
  24. Tomas CS, Nauen R (2015) IRAC, mode of action classification of insecticides resistance management. Pestic Biochem Physiol 12:122–128Google Scholar
  25. Wang H, Zhang W, Chen L, Wang JF, Liu TZ (2013) The contamination and control of biological pollutants in mass cultivation of microalgae. Bioresour Technol 128:745–750CrossRefGoogle Scholar
  26. Wang Y, Eustance E, Castillo-Keller M, Sommerfeld M (2017) Evaluation of chemical treatments for control of ciliate grazers in algae cultures: a site study. J Appl Phycol 29:2761–2770CrossRefGoogle Scholar
  27. Xie YS, Fields PG, Isman MB, Chen WK, Zhang X (1995) Insecticidal activity of Melia toosendan extracts and toosendanin against three stored-product insects. J Stored Prod Res 31:259–265CrossRefGoogle Scholar
  28. Yang Z, Kong F, Shi X, Yang J (2005) Effects of Branchionus calyciflorus culture media filtrate on Microcystis aeruginosa, Scenedesmus obliquus and Chlorella vulgaris colony formation and growth. Chin J Appl Ecol 16:1138Google Scholar
  29. Zhang YN, Ma ZQ, Wang HP, Wang ZH, Gao HY, Feng JT (2007) Evaluation of the toxicity of botanical pesticide toosendanin to non-target organisms. Acta Sci Circumst 27:2038–2045Google Scholar
  30. Zuo S, Wan K, Ma S (2015) Combined effect of predatory zooplankton and allelopathic aquatic macrophytes on algal suppression. Environ Technol 36:54–59CrossRefGoogle Scholar
  31. Zhou P, Luo L, Bai D, Li J (1987) The toosendanin induced blocking action of synaptic transmission in the 6th abdominal ganglion of cockroach, Periplaneta americana. Acta Sci Nat Univ Pekin 2:71–76Google Scholar
  32. Zhang T, Li J, Yin F, Lin B, Wang Z, Xu J, Wang H, Zuo D, Wang G, Hua Y, Cai Z (2017) Toosendanin demonstrates promising antitumor efficacy in osteosarcoma by targeting STAT3. Oncogene 36:6627–6639CrossRefGoogle Scholar

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© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.CAS Key Laboratory of Experimental Marine Biology, Institute of OceanologyChinese Academy of SciencesQingdaoChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.Laboratory for Marine Biology and BiotechnologyQingdao National Laboratory for Marine Science and TechnologyQingdaoChina

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