Acute and Synergistic Toxicity of Drugs in Water by Luminescent Bacteria Assay

Document Type : Research Paper


Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, 87036 Rende (CS), Italy


The acute toxicity in water of a series of drugs, including antibiotics, antihistamines, antifungals, steroidal and non-steroidal anti-inflammatories, was evaluated through the measurement of bioluminescence from the bacterium Vibrio fischeri. The drugs were spiked in water at concentration in the range 1.0-50.0 g/mL, distributed over six concentration levels, and their toxicity evaluated in terms of response rate calculated along 30 min of incubation. The test was also applied to real river samples previously assayed by HPLC method. The parameters LOEC (lowest observable effect concentration) and EC50 (half effective concentration) were calculated. Chlortetracycline, promethazine, betamethasone, ketoconazole and econazole were found to be very toxic. Diclofenac and ketoprofen showed toxicity only at the highest concentrations tested. Clindamycin, neomycin and oxatomide induced a decrease in bioluminescence but below the toxicity limits. In contrast, erythromycin and diphenhydramine showed an increase of bioluminescence, known as hormesis. The toxicity was amplified in samples containing drug mixtures, demonstrating additive or synergistic activity.


[1]       M.A. Boleda, M.A. Galceran, F. Ventura, Water Res. 43 (2009) 1126.
[2]       B. Kasprzyk-Hordern, R.M. Dinsdale, A.J. Guwy, Water Res. 42 (2008) 3498.
[3]       P. Vazquez-Roig, V. Andreu, C. Blasco, Y. Pico, Sci. Total Environ. 440 (2012) 24.
[4]       M. Bedner, W.A. Maccrehan, Chemosphere 65 (2006) 2130.
[5]       Y. Kim, K. Choi, J. Jung, S. Park, P.G. Kim, J. Park, Environ. Int. 33 (2007) 370.
[6]       G. Ragno, M. Veronico, C. Vetuschi, Int. J. Pharm. 99 (1993) 351.
[7]       G. Ragno, A. Risoli, G. Ioele, E. Cione, M. De Luca, J. Nanosci. Nanotechnol. 6 (2006) 2979.
[8]       M. Veronico, G. Ragno, C. Vetuschi, Spectr. Lett. 28 (1995) 407.
[9]       C. Vetuschi, G. Ragno, Int. J. Pharm. 65 (1990) 128.
[10]    A. Bianchini, C.M. Wood, Ecotoxicol. Environ. Safety 71 (2008) 32.
[11]    G.L. Brun, M. Bernier, R. Losier, K. Doe, P. Jackman, H.B. Lee, Environ. Toxicol. Chem. 25 (2006) 2163.
[12]    B. Quinn, F. Gagne, C. Blaise, The Science of the Total Environ. 389 (2008) 306.
[13]    G. Ioele, F. Oliverio, M. De Luca, G. Ragno, Curr. Pharm. Anal. 8 (2012) 196.
[14]    G. Ioele, M. De Luca, G. Ragno, Cur. Pharm. Anal. 12 (2016) 220.
[15]    X. Zou, Z. Lin, Z. Deng, D. Yin, Y. Zhang, Chemosphere, 86 (2012) 30.
[16]    S. Castiglioni, F. Pomati, K. Miller, B.P. Burns, E. Zuccato, D. Calamari, B.A. Neilan, Water Res. 42 (2008) 4271.
[17]    J.A. Zarn, B.J. Bruschweiler, J.R. Schlatter, Environ. Health Perspectives 111 (2003) 255.
[18]    Q. Huang, Y. Yu, C. Tang, X. Peng, J. Chromatogr. A 1217 (2010) 3481.
[19]    A.O. Medeiros, L.M. Kohler, J.S. Hamdan, B.S. Missagia, F.A. Barbosa, C.A. Rosa, Water Res. 42 (2008) 3921.
[20]    J.C. Van De Steene, W.E. Lambert, J. Chromatogr. A 1182 (2008) 153.
[21]    P.  Bartels,  W. von Tumpling, Jr.,  The Science of  the
Total Environ. 374 (2007) 143.
[22]    M.J. Capdeville, H. Budzinski, TrAC-Trends Anal. Chem. 30 (2011) 586.
[23]    S. Mompelat, B. Le Bot, O. Thomas, Environ. Int. 35 (2009) 803.
[24]    M. Abbas, M. Adil, S. Ehtisham-Ul-Haque, B. Munir, M. Yameen, A. Ghaffar, G.A. Shar, M. Asif Tahir, M. Iqbal, The Science of the Total Environ. 626 (2018) 1295.
[25]    E. Cotou, E. Papathanassiou, C. Tsangaris, Environ. Pollut. 119 (2002) 141.
[26]    M. Farre, D. Barcelo, TrAC-Trends Anal. Chem. 2 (2003) 299.
[27]    S. Parvez, C. Venkataraman, S. Mukherji, Environ. Int. 32 (2006) 265.
[28]    F. Conforti, G. Ioele, G.A. Statti, M. Marrelli, G. Ragno, F. Menichini, An International Journal Published for the British Industrial Biological Research Association 46 (2008) 3325.
[29]    E. Fulladosa, J.C. Murat, M. Martinez, I. Villaescusa, Arch. Environ. Contamination and Toxicol. 46 (2004) 176.
[30]    J.M. Ribò, K.L. Kaiser, Environ. Toxicol. 2 (2006).
[31]    E.J. Calabrese, L.A.  Baldwin,  Critical  Rev.  Toxicol.
31 (2001) 353.
[32]    D.A. Dawson, G. Poch, T.W. Schultz, Ecotoxicol. Environ. Safety 65 (2006) 171.
[33]    I. Rodea-Palomares, A.L. Petre, K. Boltes, F. Leganésa, J.A. Perdigón-Melón, R. Rosal, F. Water Res. 44 (2010) 427.
[34]    I. Italy, Determination of the Inhibitory Effect of Water Samples on the Light Emission of Vibrio fischeri, 2007. deleghe/06152dl.htm, in.
[35]    R.R. Azur, Carlsbad, CA., Microtox Manual, Azur Environmental (Formely Microbics Corporation), 1997.
[36]    I. Irsa, Metodi Analitici per le Acque. Quadro Istituzionale Ricerca Acque, 1995.
[37]    C.W. Dunnett, A multiple comparison procedure for comparing several treatments with a control, J. Am. Stat. Assoc. 50 (1955) 1096.
[38]    L. Koshy, T. Jones, K. BeruBe, Bioreactivity of municipal solid waste landfill leachates-Hormesis and DNA damage, Water Res. 42 (2008) 2177.
[39]    V.D.S. Nunes-Halldorson, N.L. Duran, Bioluminescent bacteria: Lux genes as environmental biosensors, Brazilian J.  Microbiol. 34 (2003).