Environmental concerns about the effects of effluents from wastewater treatment plants in tourist areas of the Alps: toxicity in aquatic microorganisms
Are the effluents of wastewater treatment plants in high mountains of concern for aquatic biodiversity? To answer this question, we carried out an experimental study testing the short-term toxicity of some Pharmaceutical Active Compounds (PhACs) in the effluents of a plant in a mountain valley of the Italian Alps sampled during the high tourist season (i.e., the ski season) when PhACs contamination is higher. We used different tools, taking as a model the bacterium Aliivibrio fischeri: the “whole-mixture approach” (Microtox test), “component-based approach”, predictive models “Concentration Addition (CA)”, “Independent Action (IA)”, and Combination Index (CI)”. We investigated the nature of interactions potentially occurring among seven selected PhACs (clarithromycin, naproxen, acetaminophen (paracetamol), ibuprofen, diclofenac, carbamazepine, and amoxicillin). This study showed that anti-inflammatory ibuprofen and diclofenac have higher short-term toxicity (IC50 <100 mg L-1) for A. fischeri compared with antibiotics, whose toxic effects are expected to become visible in the long term. Furthermore, based on the CI method, the seven PhACs seem not to interact in a synergistic or antagonistic way, but the final effect of their mixture seems to be equal to the sum of their individual effects. Notwithstanding the high tourist pressure, the Microtox test reported an overall toxicity of only 21%, which drops to 7% in the receiving water body, the Vermigliana stream. These values, besides the predictions by CA and IA, are not alarming per se, i.e., the treated effluent of the plant in the period of maximum tourist pressure can be considered no harmful to aquatic microorganisms. However, based on other studies highlighting negative effects of the diluted treated effluent of the same plant on macroinvertebrate community structure, we suggest that other model organisms be considered, including algae, insects, and fish, to assess the real ecological risk to wildlife of an effluent. The experimental tests on A. fischeri are useful for fast, preliminary information on the level of risk for freshwater ecosystems, but they should be combined with field studies and experiments on the wild populations to increase the ecological realism.
ADEP, Agenzia per la Depurazione. Available from https://adep.provincia.tn.it/Agenzia-perla-Depurazione-ADEP
Alpine Convention, 2009. Water and water management issues: Report on the state of the Alps. Alpine signals. Special edition p. 2. Innsbruck, Austria: Permanent Secretariat of the Alpine Convention.
Azur Environmental, 1998. Microtox System Operating manual. Azur Environmental, Carlsbad.
Backhaus T., Grimme LH, 1999. The toxicity of antibiotic agents to the luminescent bacterium Vibrio fischeri. Chemosphere 38:3291-3301. DOI: https://doi.org/10.1016/S0045-6535(98)00560-8
Backhaus T, Karlsson M, 2014. Screening level mixture risk assessment of pharmaceuticals in STP effluents. Water Res. 49:157-165. DOI: https://doi.org/10.1016/j.watres.2013.11.005
Backhaus T, 2014. Medicines, shaken and stirred: a critical review on the ecotoxicology of pharmaceutical mixtures. Phil. Trans. R. Soc. B 369:20130585. DOI: https://doi.org/10.1098/rstb.2013.0585
Berenbaum MC, 1985. The expected effect of a combination of agents: the general solution. J. Theor. 114:413-31. DOI: https://doi.org/10.1016/S0022-5193(85)80176-4
Belden JB, Gilliom RJ, Lydy MJ, 2007. How well can we predict the toxicity of pesticide mixtures to aquatic life? Integr. Environ. Assess. Manag. 3:364e372. DOI: https://doi.org/10.1002/ieam.5630030307
Biel-Maeso M, Corada-Fernández C, Lara-Martín PA, 2019. Removal of personal care products (PCPs) in wastewater and sludge treatment and their occurrence in receiving soils. Water Res. 150:129-139. DOI: https://doi.org/10.1016/j.watres.2018.11.045
Bliss CI, 1939. The toxicity of poisons applied jointly. Ann. Appl. Biol. 26:585-615. DOI: https://doi.org/10.1111/j.1744-7348.1939.tb06990.x
Buffagni A, Erba S, 2008. [Definizione dello stato ecologico dei fiumi sulla base dei macroinvertebrati bentonici per la 2000/60/CE(WFD): il sistema di classificazione MacrOper].[in Italian]. IRSA–CNR, Notiziario dei Metodi Analitici: 24-46.
Carballa M, Omil F, Lema JM, 2005. Removal of cosmetic ingredients and pharmaceuticals in sewage primary treatment. Water Res. 39:4790-4796. DOI: https://doi.org/10.1016/j.watres.2005.09.018
Cedergreen N, 2014. Quantifying synergy: a systematic review of mixture toxicity studies within environmental toxicology. PLoS One 9:e96580. DOI: https://doi.org/10.1371/journal.pone.0096580
Chiogna G, Majone B, Cano Paoli K, Diamantini E, Mallucci S, Stella E, Lencioni V, Zandonai F, Bellin A, 2016. A review of hydrological and chemical stressors in the Adige catchment and its ecological status. Sci. Total Environ. 540:429-443. DOI: https://doi.org/10.1016/j.scitotenv.2015.06.149
Chou TC, Talalay P, 1983. Analysis of combined drug effects: a new look at a very old problem. Trends Pharmacol. Sci. 4:450-454. DOI: https://doi.org/10.1016/0165-6147(83)90490-X
Chou TC, 1976. Derivation and properties of Michaelis-Menten type and Hill type equations for reference ligands. J. Theor. Biol. 59:253-254. DOI: https://doi.org/10.1016/0022-5193(76)90169-7
Chou TC, 2006. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol. Rev. 58:621-681. DOI: https://doi.org/10.1124/pr.58.3.10
Chou TC, Martin N, 2005. CompuSyn for Drug Combinations: PC Software and User’s Guide: A Computer Program for Quantitation of Synergism and Antagonism in Drug Combinations, and the Determination of IC50 and ED50 and LD50 Values. ComboSyn Inc, Paramus.
Chou TC, Talalay P, 1984. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv. Enzym. Regul. 22:27-55. DOI: https://doi.org/10.1016/0065-2571(84)90007-4
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) 12:12741–12771. DOI: https://doi.org/10.3390/s120912741
Daughton CG, Ternes TA, 1999. Pharmaceuticals and personal care products in the environment: agents of subtle change? Environ. Health Perspect. 107:s907-938. DOI: https://doi.org/10.1289/ehp.99107s6907
Daughton CG, 2005. "Emerging" chemicals as pollutants in the environment: A 21st century perspective. Renew. Resour. J. 23:6-23.
Deprez K, Robbens J, Nobels I, Vanparys C, Vanermen G, Tirez K, Michiels L, Weltens R, 2012. DISCRISET: A battery of tests for fast waste classification - Application of tests on waste extracts. Waste Manage. 32: 2218-2228. DOI: https://doi.org/10.1016/j.wasman.2012.05.017
Di Nica V, Villa S, Finizio A, 2016. Experimental and predicted toxicity of binary combinations of diclofenac sodium, carbamazepine and caffeine to Aliivibrio fischeri. Environ. Eng. Manag. J. 15:1971-1980. DOI: https://doi.org/10.30638/eemj.2016.212
Di Nica V, Gallet J, Villa S, Mezzanotte V, 2017a. Toxicity of quaternary ammonium compounds (QACs) as single compounds and mixtures to aquatic non-target microorganisms: experimental data and predictive models. Ecotoxicol. Environ. Saf. 142:567-577. DOI: https://doi.org/10.1016/j.ecoenv.2017.04.028
Di Nica V, Villa S., Finizio A, 2017b. Toxicity of individual pharmaceuticals and their mixtures to Aliivibrio fischeri: evidence of toxicological interactions in binary combinations. Environ. Toxicol. Chem. 36:815-822. DOI: https://doi.org/10.1002/etc.3686
Di Nica V, Villa S, Finizio A, 2017c. Toxicity of individual pharmaceuticals and their mixtures to Aliivibrio fischeri: experimental results for single compounds and considerations of their mechanisms of action and potential acute effects on aquatic organisms. Environ. Toxicol. Chem. 36:807-814. DOI: https://doi.org/10.1002/etc.3568
DIN EN ISO 11348-2, 2007. Water quality - Determination of the inhibitory effect of water samples on the light emission of Vibrio fischeri (Luminescent bacteria test) Method using liquid-dried bacteria.
Dong Y, Wang J, Ding L, Liu Y, 2013. Influence of cosolvents on low water-solubility chemicals to Photobacterium phosphoreum in acute toxicity test. Procedia Environ. Sci. 18:143-148. DOI: https://doi.org/10.1016/j.proenv.2013.04.019
Escher BI, Baumer A, Bittermann K, Henneberger L, Konig M, Kuhnert C, Kluver N, 2017. General baseline toxicity QSAR for nonpolar, polar and ionisable chemicals and their mixtures in the bioluminescence inhibition assay with Aliivibrio fischeri. Environ. Sci. 19:414-428. DOI: https://doi.org/10.1039/C6EM00692B
European Commission, 2000. Directive 2000/60/EC. Establishing a framework for community action in the field of water policy. European Commission PE-Cons 3639/1/100 Rev 1. Commission to the European Communities, Luxembourg City, Luxembourg.
González-Pleiter M, Gonzalo S, Rodea-Palomares I, Leganés F, Rosal R, Boltes K, Marco E, Fernández-Piñas F, 2013. Toxicity of five antibiotics and their mixtures towards photosynthetic aquatic organisms: implications for environmental risk assessment. Water Res. 47:2050-64. DOI: https://doi.org/10.1016/j.watres.2013.01.020
Gosset A, Wiest L, Fildier A, Libert C, Giroud B, Hammada M, Hervé M, Sibeud E, Vulliet E, Polomé P, Perrodin Y, 2021. Ecotoxicological risk assessment of contaminants of emerging concern identified by “suspect screening” from urban wastewater treatment plant effluents at a territorial scale. Sci. Total Environ. 778:146275. DOI: https://doi.org/10.1016/j.scitotenv.2021.146275
Gosset A, Polomé P, Perrodin Y, 2020. Ecotoxicological risk assessment of micropollutants from treated urban wastewater effluents for watercourses at a territorial scale: Application and comparison of two approaches. Int. J. Hyg. Envir. Heal. 224:113437. DOI: https://doi.org/10.1016/j.ijheh.2019.113437
Groten JP, Feron VJ, Suhnel J, 2001. Toxicology of simple and complex mixtures. Trends Pharm. Sci. 22:316-322. DOI: https://doi.org/10.1016/S0165-6147(00)01720-X
IRSA-CNR, APAT, 2003. [Metodi analitici per le acque. Manuali e Linee guida].[in Italian]. IRSA-CNR, APAT.
Kim Y, Choi K, Jung J, Park S, Kim P-G, Park J, 2007. Aquatic toxicity of acetaminophen, carbamazepine, cimetidine, diltiazem and six major sulfonamides, and their potential ecological risks in Korea. Environ. Int. 33:370-375. DOI: https://doi.org/10.1016/j.envint.2006.11.017
Kienzler A, Berggren E, Bessems J, Bopp S, van der Linden S, Worth A, 2014. Assessment of mixtures - review of regulatory requirements and guidance. JRC Science for Policy Report, EUR 26675EN. Luxembourg: Publications Office of the European Union.
Kortenkamp A, Backhaus T, Faust M, 2009. State of the art report on mixture toxicity. Final report to the European Commission. Contract no. 070307/2007/485103/ETU/D.1.
Lencioni V, Bellamoli F, Paoli F, 2020. Multi-level effects of emerging contaminants on macroinvertebrates in Alpine streams: from DNA to the ecosystem. Ecol. Indic. 117:106660. DOI: https://doi.org/10.1016/j.ecolind.2020.106660
Loewe S, Muischne KH, 1926. Effect of combinations: Mathematical basis of problem. N-S Arch. Exp. Pathol. Physiol. 114:313-326. DOI: https://doi.org/10.1007/BF01952257
Mandaric L, Diamantini E, Stella E, Cano-Paoli K, Valle-Sistac J, Molins-Delgado D, Bellin A, Chiogna G, Majone B, Diaz-Cruz MS, Sabater S, Barcelo D, Petrovic M, 2017. Contamination sources and distribution patterns of pharmaceuticals and personal care products in Alpine rivers strongly affected by tourism. Sci. Total Environ. 590-591:484-494. DOI: https://doi.org/10.1016/j.scitotenv.2017.02.185
Monteiro SC, Boxall ABA, 2010. Occurrence and fate of human pharmaceuticals in the environment. Rev Environ Contam Toxicol. 202:53-154. DOI: https://doi.org/10.1007/978-1-4419-1157-5_2
OECD, 2000. Series on Testing and Assessment No. 23. Guidance document on aquatic toxicity testing of difficult substances and mixtures. Environment Directorate, Organisation For Economic Co-Operation And Development, Paris. ENV/JM/MONO(2000)6.
O'Flynn D, Lawler J, Yusuf A, Parle-McDermott A, Harold D, Mc Cloughlin T, Holland L, Regan F, Blanaid W, 2021. A review of pharmaceutical occurrence and pathways in the aquatic environment in the context of a changing climate and the COVID-19 pandemic. Anal. Methods 13:575. DOI: https://doi.org/10.1039/D0AY02098B
Ortiz de García SA, Pinto Pinto G, García-Encina PA, Irusta-Mata R, 2014. Ecotoxicity and environmental risk assessment of pharmaceuticals and personal care products in aquatic environments and wastewater treatment plants. Ecotoxicology 23:1517-1533. DOI: https://doi.org/10.1007/s10646-014-1293-8
Parvez S, Venkataraman C, Mukherji S, 2006. A review on advantages of implementing luminescence inhibition test (Vibrio fischeri) for acute toxicity prediction of chemicals. Environ. Int. 32:265-268. DOI: https://doi.org/10.1016/j.envint.2005.08.022
Price P, Han X, Junghans M, Kunz P, Watts C, Leverett D, 2012. An application of a decision tree for assessing effects from exposures to multiple substances to the assessment of human and ecological effects from combined exposures to chemicals observed in surface waters and wastewater effluents. Environ. Sci. Eur. 24:1-13. DOI: https://doi.org/10.1186/2190-4715-24-34
R Core Team. 2021. R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria. Available from: http://www.R-project.org/
Rodea-Palomares I, Petre AL, Boltes K, Leganés F, Perdigón-Melón JA, Rosal R, Fernández-Piñas F, 2010. Application of the combination index (CI)-isobologram equation to study the toxicological interactions of lipid regulators in two aquatic bioluminescent organisms. Water Res. 44:427-438. DOI: https://doi.org/10.1016/j.watres.2009.07.026
SCHER, SCCS, SCENIHR, Opinion on the Toxicity and Assessment of Chemical Mixtures, 2012.
Silva E, Rajapakse N, Kortenkamp A, 2002. Something From “Nothing”-Eight Weak Estrogenic Chemicals Combined at Concentrations Below NOECs Produce Significant Mixture Effects. Environ. Sci. Technol. 36:1751-1756. DOI: https://doi.org/10.1021/es0101227
Sprague JB, 1970. Measurement of pollutant toxicity to fish. II. Utilizing and applying bioassay results. Water Res. 4:3-32. DOI: https://doi.org/10.1016/0043-1354(70)90018-7
Thrupp TJ, Runnalls TJ, Scholze M, Kugathas S, Kortenkamp A, Sumpter JP, 2018. The consequences of exposure to mixtures of chemicals: Something from ‘nothing’ and ‘a lot from a little’ when fish are exposed to steroid hormones. Sci. Total Environ. 619-620:1482-1492. DOI: https://doi.org/10.1016/j.scitotenv.2017.11.081
United Nations, 2013. Globally Harmonized System of Classification and Labelling of Chemicals, 5th ed. United Nations Publications. ST/SG/AC. 10/30/Rev.5. New York, NY.
van Gestel CAM, Jonker MJ, Kammenga JE, Laskowski R, Svendsen C, 2011. Mixture toxicity: linking approaches from ecological and human toxicology. CRC Press, Boca Raton.
Verhaar HJM, Van Leeuwen CJ, Hermens JLM, 1992. Classifying environmental pollutants. 1: Structure-activity relationships for prediction of aquatic toxicity. Chemosphere 25:471-491. DOI: https://doi.org/10.1016/0045-6535(92)90280-5
Vighi M, Migliorati S, Monti GS 2009. Toxicity on the luminescent bacterium Vibrio fischeri (Beijerinck). I: QSAR equation for narcotics and polar narcotics. Ecotoxicol. Environ. Saf. 72:154-161. DOI: https://doi.org/10.1016/j.ecoenv.2008.05.008
Villa S, Di Nica V, Castiglioni S, Finizio A, 2020. Environmental risk classification of emerging contaminants in an alpine stream influenced by seasonal tourism. Ecol. Indic. 115:06428. DOI: https://doi.org/10.1016/j.ecolind.2020.106428
Villa S, Di Nica V, Bellamoli F, Pescatore T, Ferrario C, Finizio A, Lencioni V, 2018a. Effects of a treated sewage effluent on behavioural traits in Diamesa cinerella and Daphnia magna. J. Limnol. 77:1760. DOI: https://doi.org/10.4081/jlimnol.2018.1760
Villa S, Di Nica V, Pescatore T, Bellamoli F, Miari F, Finizio A, Lencioni V, 2018b. Comparison of the behavioural effects of pharmaceuticals and pesticides on Diamesa zernyi larvae (Chironomidae). Env. Poll. 238:130-139. DOI: https://doi.org/10.1016/j.envpol.2018.03.029
Warne MStJ, Hawker DW, 1995. The number of components in a mixture determines whether synergistic and antagonistic or additive toxicity predominate: The funnel hypothesis. Ecotoxicol. Environ. Saf. 31:23-28. DOI: https://doi.org/10.1006/eesa.1995.1039
Weltens R, Vanermen G, Tirez K, Robbens J, Deprez K, Michiels L, 2012. Screening tests for hazard classification of complex waste materials – Selection of methods, Waste Manag. 32:2208-2217. DOI: https://doi.org/10.1016/j.wasman.2012.05.013
Weltens R, Deprez K, Michiels L, 2014. Validation of Microtox as a first screening tool for waste classification. Waste Manag. 34:2427-2433. DOI: https://doi.org/10.1016/j.wasman.2014.08.001
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