Impact of Suspended Solids on the Use of LuminoTox to Detect Toxicity of Micropollutants

  • Meghan Marshall
  • Marco Pineda
  • Viviane Yargeau


There is an increasing need for tools to monitor toxicity of contaminants of emerging concern (CECs) in wastewater. The purpose of this work was to assess interferences in the presence of total solids (TS) and total suspended solids (TSS) in the LuminoTox at concentrations typical of those found in municipal secondary effluent (SE) and to evaluate a simple sample enrichment method for increased CEC sensitivity. 4 or 10 µg/L atrazine in different TS concentrations and in corresponding filtrates (TSS removed) exhibited equivalent toxicities. Because the only difference between these two fractions is the TSS, this result demonstrates that, generally, this fraction does not induce toxicity nor interfere with the bioassay. At constant medium-low TS, the LuminoTox was able to detect the presence of 4 µg/L of atrazine but could not distinguish the change in atrazine concentration between 4 and 6 µg/L. No inhibition was observed in the presence of a mix of 14 CECs each at 0.23 µg/L. However, upon sample enrichment by lyophilization (50×), an inhibition of 81 ± 3% was observed. The enriched SE alone (not spiked with CECs) led to an inhibition of 49 ± 1%, indicating the detection of the CEC contribution to toxicity after sample preconcentration. The LuminoTox is a promising tool for monitoring SE; however, if the intent is to detect CECs, enrichment method optimization is required.



Stabilized aqueous photosynthetic systems


Contaminants of emerging concern


Secondary effluent


Synthetic wastewater


Milli Q water




Photosystem II


Natural Organic Matter


Humic acids


Particle size distribution


LuminoTox Solid Phase Assay


LuminoTox Leachate Phase Assay


LuminoTox Direct Contact Assay



The authors thank the staff of each WWTP for their cooperation and help during sampling campaigns. They thank Marco Pineda for his assistance with chemical analysis of the target CECs and Andrew Golsztajn and Ranjan Roy for their help with the Particle Size Distribution Analysis. The authors also thank the McGill Engineering Doctoral Award for supporting Meghan Marshall. A research grant was provided to Viviane Yargeau (PI) from the Natural Sciences and Engineering Research Council (NSERC) of Canada through the Discovery Grant Program (RGPIN/04635-2015).

Supplementary material

244_2017_478_MOESM1_ESM.docx (16 kb)
Supplementary material 1 (DOCX 15 kb)


  1. Altenburger R, Backhaus T, Boedeker W et al (2013) Simplifying complexity: mixture toxicity assessment in the last 20 years. Environ Toxicol Chem 32(8):1685–1687. CrossRefGoogle Scholar
  2. Baek S-O, Chang I-S (2009) Pretreatments to control membrane fouling in membrane filtration of secondary effluents. Desalination 244(1–3):153–163. CrossRefGoogle Scholar
  3. Bellemare F, Rouette M-E, Lorrain L et al (2006) Combined use of photosynthetic enzyme complexes and microalgal photosynthetic systems for rapid screening of wastewater toxicity. Environ Toxicol 21(5):445–449. CrossRefGoogle Scholar
  4. Boltes K, Rosal R, García-Calvo E (2012) Toxicity of mixtures of perfluorooctane sulphonic acid with chlorinated chemicals and lipid regulators. Chemosphere 86(1):24–29. CrossRefGoogle Scholar
  5. Boucher N, Carpentier R (1999) Hg2+, Cu2+, and Pb2+-induced changes in Photosystem II photochemical yield and energy storage in isolated thylakoid membranes: a study using simultaneous fluorescence and photoacoustic measurements. Photosynth Res 59(2):167–174. CrossRefGoogle Scholar
  6. Burga Pérez KF, Charlatchka R, Férard J-F (2013) Assessment of the LuminoTox leachate phase assay as a complement to the LuminoTox solid phase assay: effect of fine particles in natural sediments. Chemosphere 90(3):1310–1315. CrossRefGoogle Scholar
  7. Buth JM, Ross MR, McNeill K, Arnold WA (2011) Reprint of: removal and formation of chlorinated triclosan derivatives in wastewater treatment plants using chlorine and UV disinfection. Chemosphere 85(2):284–289. CrossRefGoogle Scholar
  8. Carlson JC, Anderson JC, Low JE et al (2013) Presence and hazards of nutrients and emerging organic micropollutants from sewage lagoon discharges into Dead Horse Creek, Manitoba, Canada. Sci Total Environ 445–446:64–78. CrossRefGoogle Scholar
  9. Cicek N, Londry K, Oleszkiewicz JA et al (2007) Removal of selected natural and synthetic estrogenic compounds in a Canadian full-scale municipal wastewater treatment plant. Water Environ Res 79(7):795–800. CrossRefGoogle Scholar
  10. Connon RE, Geist J, Werner I (2012) Effect-based tools for monitoring and predicting the ecotoxicological effects of chemicals in the aquatic environment. Sensors (Basel, Switzerland) 12(9):12741–12771. CrossRefGoogle Scholar
  11. Conrad R, Büchel C, Wilhelm C et al (1993) Changes in yield ofin-vivo fluorescence of chlorophyll a as a tool for selective herbicide monitoring. J Appl Phycol 5(5):505–516. CrossRefGoogle Scholar
  12. Dellamatrice PM, Monteiro RTR, Blaise C et al (2006) Toxicity assessment of reference and natural freshwater sediments with the LuminoTox assay. Environ Toxicol 21(4):395–402. CrossRefGoogle Scholar
  13. Dewez D, Boucher N, Bellemare F, Popovic R (2007) Use of different fluorometric systems in the determination of fluorescence parameters from spinach thylakoid membranes being exposed to atrazine and copper. Toxicol Environ Chem 89(4):655–664. CrossRefGoogle Scholar
  14. Environment Canada (2002) Biological test method: solid-phase reference method for determining the toxicity of sediment using luminescent bacteria (Vibrio fischeri). Environ Protect Ser Report EPS 1/RM/42. Retrieved from Ottawa ONGoogle Scholar
  15. Férard J, Burga Pérez K, Blaise C et al (2015) Microscale ecotoxicity testing of Moselle river watershed (Lorraine Province, France) sediments. J Xenobiotics 5(1):1–7. CrossRefGoogle Scholar
  16. Fernandez MP, Ikonomou MG, Buchanan I (2007) An assessment of estrogenic organic contaminants in Canadian wastewaters. Sci Total Environ 373(1):250–269. CrossRefGoogle Scholar
  17. Harris GD, Adams VD, Sorensen DL, Dupont RR (1987) The influence of photoreactivation and water quality on ultraviolet disinfection of secondary municipal wastewater. J Water Pollut Control Fed 59(8):781–787Google Scholar
  18. Hiriart-Baer VP, Fortin C, Lee D-Y, Campbell PGC (2006) Toxicity of silver to two freshwater algae, Chlamydomonas reinhardtii and Pseudokirchneriella subcapitata, grown under continuous culture conditions: influence of thiosulphate. Aquat Toxicol 78(2):136–148. CrossRefGoogle Scholar
  19. Hua WY, Bennett ER, Maio X-S et al (2006) Seasonality effects on pharmaceuticals and s-triazine herbicides in wastewater effluent and surface water from the Canadian side of the upper Detroit River. Environ Toxicol Chem 25(9):2356–2365. CrossRefGoogle Scholar
  20. Jasinska EJ, Goss GG, Gillis PL et al (2015) Assessment of biomarkers for contaminants of emerging concern on aquatic organisms downstream of a municipal wastewater discharge. Sci Total Environ 530–531:140–153. CrossRefGoogle Scholar
  21. Jonker MJ, Svendsen C, Bedaux JJM et al (2005) Significance testing of synergistic/antagonistic, dose level-dependent, or dose ratio-dependent effects in mixture dose-response analysis. Environ Toxicol Chem 24(10):2701–2713. CrossRefGoogle Scholar
  22. Kerr JL, Guo Z, Smith DW et al (2008) Use of goldfish to monitor wastewater and reuse water for xenobiotics. J Environ Eng Sci 7(4):369–383. CrossRefGoogle Scholar
  23. Krewski D, Acosta D Jr, Andersen M et al (2010) Toxicity testing in the 21st century: a vision and a strategy. J Toxicol Environ Health B Crit Rev 13(2–4):51–138. CrossRefGoogle Scholar
  24. Lajeunesse A, Smyth SA, Barclay K et al (2012) Distribution of antidepressant residues in wastewater and biosolids following different treatment processes by municipal wastewater treatment plants in Canada. Water Res 46(17):5600–5612. CrossRefGoogle Scholar
  25. Levine AD, Tchobanoglous G, Asano T (1985) Characterization of the size distribution of contaminants in wastewater: treatment and reuse implications. J Water Pollut Control Fed 57(7):805–816.
  26. Lishman L, Smyth SA, Sarafin K et al (2006) Occurrence and reductions of pharmaceuticals and personal care products and estrogens by municipal wastewater treatment plants in Ontario, Canada. Sci Total Environ 367(2–3):544–558. CrossRefGoogle Scholar
  27. Lonappan L, Brar SK, Das RK et al (2016) Diclofenac and its transformation products: environmental occurrence and toxicity: a review. Environ Int 96:127–138. CrossRefGoogle Scholar
  28. Macedo RS, Lombardi AT, Omachi CY, Rörig LR (2008) Effects of the herbicide bentazon on growth and photosystem II maximum quantum yield of the marine diatom Skeletonema costatum. Toxicol In Vitro 22(3):716–722. CrossRefGoogle Scholar
  29. Maksymiec W, Baszyński T (1988) The effect of Cd2+ on the release of proteins from thylakoid membranes of tomato leaves. Acta Soc Bot Pol 57(4):465–474. CrossRefGoogle Scholar
  30. Mamindy-Pajany Y, Hamer B, Roméo M et al (2011) The toxicity of composted sediments from Mediterranean ports evaluated by several bioassays. Chemosphere 82(3):362–369. CrossRefGoogle Scholar
  31. Marshall M, Yargeau V (2017) Sensitivity of the LuminoTox tool to monitor contaminants of emerging concern in municipal secondary wastewater effluent. Sci Total Environ 598:1065–1075. CrossRefGoogle Scholar
  32. Maruya KA, Dodder NG, Mehinto AC et al (2016) A tiered, integrated biological and chemical monitoring framework for contaminants of emerging concern in aquatic ecosystems. Integr Environ Assess Manag 12(3):540–547. CrossRefGoogle Scholar
  33. Metcalfe CD, Koenig BG, Bennie DT et al (2003) Occurrence of neutral and acidic drugs in the effluents of Canadian sewage treatment plants. Environ Toxicol Chem 22(12):2872–2880. CrossRefGoogle Scholar
  34. Metcalfe CD, Chu S, Judt C et al (2010) Antidepressants and their metabolites in municipal wastewater, and downstream exposure in an urban watershed. Environ Toxicol Chem 29(1):79–89. CrossRefGoogle Scholar
  35. Metcalfe CD, Kleywegt S, Letcher RJ et al (2013) A multi-assay screening approach for assessment of endocrine-active contaminants in wastewater effluent samples. Sci Total Environ 454–455:132–140. CrossRefGoogle Scholar
  36. Miao XS, Bishay F, Chen M, Metcalfe CD (2004) Occurrence of antimicrobials in the final effluents of wastewater treatment plants in Canada. Environ Sci Technol 38(13):3533–3541. CrossRefGoogle Scholar
  37. Muller R, Schreiber U, Escher BI et al (2008) Rapid exposure assessment of PSII herbicides in surface water using a novel chlorophyll a fluorescence imaging assay. Sci Total Environ 401(1–3):51–59. CrossRefGoogle Scholar
  38. Neale PA, Antony A, Bartkow ME et al (2012) Bioanalytical assessment of the formation of disinfection byproducts in a drinking water treatment plant. Environ Sci Technol 46(18):10317–10325. Google Scholar
  39. Osorio V, Larrañaga A, Aceña J et al (2016) Concentration and risk of pharmaceuticals in freshwater systems are related to the population density and the livestock units in Iberian Rivers. Sci Total Environ 540:267–277. CrossRefGoogle Scholar
  40. Pan Y, Zhang X, Wagner ED et al (2014) Boiling of simulated tap water: effect on polar brominated disinfection byproducts, halogen speciation, and cytotoxicity. Environ Sci Technol 48(1):149–156. CrossRefGoogle Scholar
  41. Péry ARR, Babut MP, Mons R, Garric J (2006) Deriving effects on Chironomus population carrying capacity from standard toxicity tests. Environ Toxicol Chem 25(1):144–148. CrossRefGoogle Scholar
  42. Pflugmacher S, Pietsch C, Rieger W, Steinberg CEW (2006) Dissolved natural organic matter (NOM) impacts photosynthetic oxygen production and electron transport in coontail Ceratophyllum demersum. Sci Total Environ 357(1–3):169–175. CrossRefGoogle Scholar
  43. Plewa MJ, Wagner ED, Metz DH et al (2012) Differential toxicity of drinking water disinfected with combinations of ultraviolet radiation and chlorine. Environ Sci Technol 46(14):7811–7817. CrossRefGoogle Scholar
  44. Prasse C, Stalter D, Schulte-Oehlmann U et al (2015) Spoilt for choice: a critical review on the chemical and biological assessment of current wastewater treatment technologies. Water Res 87:237–270. CrossRefGoogle Scholar
  45. Ragush CM, Schmidt JJ, Krkosek WH et al (2015) Performance of municipal waste stabilization ponds in the Canadian Arctic. Ecol Eng 83:413–421. CrossRefGoogle Scholar
  46. Rai LC, Gaur JP, Kumar HD (1981) Protective effects of certain environmental factors on the toxicity of zinc, mercury, and methylmercury to Chlorella vulgaris. Environ Res 25(2):250–259. CrossRefGoogle Scholar
  47. Rai LC, Singh AK, Mallick N (1991) Studies on photosynthesis, the associated electron transport system and some physiological variables of Chlorella vulgaris under heavy metal stress. J Plant Physiol 137(4):419–424. CrossRefGoogle Scholar
  48. Rice EW, Baird RB, Eaton AD, Clesceri LS (2012). Standard methods for the examination of water and wastewater, 22 edn. American Public Health Association, American Water Works Association, Water Environment Federation, pp. 5:21–25:25Google Scholar
  49. Ringwood AH, DeLorenzo ME, Ross PE, Holland AF (1997) Interpretation of Microtox® solid-phase toxicity tests: the effects of sediment composition. Environ Toxicol Chem 16:1135–1140. CrossRefGoogle Scholar
  50. Rojas MR, Leung C, Bonk F et al (2013) Assessment of the effectiveness of secondary wastewater treatment technologies to remove trace chemicals of emerging concern. Crit Rev Environ Sci Technol 43(12):1281–1314. CrossRefGoogle Scholar
  51. Rusten B, McCoy M et al (1998) The innovative moving bed biofilm reactor/solids contact reaeration process for secondary treatment of municipal wastewater. Water Environ Res 70(5):1083–1089.
  52. Sengupta A, Lyons JM, Smith DJ et al (2014) The occurrence and fate of chemicals of emerging concern in coastal urban rivers receiving discharge of treated municipal wastewater effluent. Environ Toxicol Chem 33(2):350–358. CrossRefGoogle Scholar
  53. Servos MR, Bennie DT, Burnison BK et al (2005) Distribution of estrogens, 17β-estradiol and estrone, in Canadian municipal wastewater treatment plants. Sci Total Environ 336(1–3):155–170. CrossRefGoogle Scholar
  54. Shaw M, Negri A, Fabricius K, Mueller JF (2009) Predicting water toxicity: pairing passive sampling with bioassays on the Great Barrier Reef. Aquat Toxicol 95(2):108–116. CrossRefGoogle Scholar
  55. Shon HK, Vigneswaran S, Snyder SA (2006) Effluent organic matter (EfOM) in wastewater: constituents, effects, and treatment. Crit Rev Environ Sci Technol 36(4):327–374. CrossRefGoogle Scholar
  56. Souza BS, Dantas RF, Agulló-Barceló M et al (2013) Evaluation of UV/H2O2 for the disinfection and treatment of municipal secondary effluents for water reuse. J Chem Technol Biotechnol 88(9):1697–1706. CrossRefGoogle Scholar
  57. Speth TF, Miltner RJ, Richardson SD, Simmons JE (2008) Integrated disinfection by-products mixtures research: concentration by reverse osmosis membrane techniques of disinfection by-products from water disinfected by chlorination and ozonation/postchlorination. J Toxicol Environ Health A 71(17):1149–1164. CrossRefGoogle Scholar
  58. Stalter D, Peters LI, O’Malley E et al (2016) Sample enrichment for bioanalytical assessment of disinfected drinking water: concentrating the polar, the volatiles, and the unknowns. Environ Sci Technol 50(12):6495–6505. CrossRefGoogle Scholar
  59. Steinberg CEW, Paul A, Pflugmacher S et al (2003) Pure humic acid substances have the potential to act as xenobiotic chemicals: a review. Fresenius Environ Bull 12(5):391–401.
  60. Tay KL, Doe KG, MacDonald AJ, Lee K (1998) Microscale testing in aquatic toxicology: advances, techniques and practice. In: Wells PG, Lee K, Blaise C (eds) The influence of particle size, ammonia and sulfide on toxicity of dredged materials for ocean disposal. CRC Press, New YorkGoogle Scholar
  61. Uslu MO, Jasim S, Arvai A et al (2013) A survey of occurrence and risk assessment of pharmaceutical substances in the Great Lakes Basin. Ozone Sci Eng 35(4):249–262. CrossRefGoogle Scholar
  62. Wang S, Zhang X, Wang Z et al (2014) In-depth characterization of secondary effluent from a municipal wastewater treatment plant located in northern China for advanced treatment. Water Sci Technol 69(7):1482–1488. CrossRefGoogle Scholar
  63. Winner RW, Owen HA (1991) Toxicity of copper to Chlamydomonas reinhardtii (Chlorophyceae) and Ceriodaphnia dubia (Crustacea) in relation to changes in water chemistry of a freshwater pond. Aquat Toxicol 21(3):157–169. CrossRefGoogle Scholar
  64. Yao Q, Wang X, Jian H et al (2015) Characterization of the particle size fraction associated with heavy metals in suspended sediments of the Yellow River. Int J Environ Res Public Health 12(6):6725–6744. CrossRefGoogle Scholar
  65. Yeh RYL, Farré MJ, Stalter D et al (2014) Bioanalytical and chemical evaluation of disinfection by-products in swimming pool water. Water Res 59:172–184. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Chemical EngineeringMcGill UniversityMontrealCanada

Personalised recommendations