A Toxicological Assessment of Endocrine Disrupting Chemicals Found in the BMW (Border, Midland and Western) Region of Ireland


A battery of tests was established to determine the oestrogenic, mutagenic and genotoxic potential of two categories of endocrine disrupting chemicals (EDCs), phthalates and alkylphenols. Diisononylphthalate (DINP), diethylhexylphthalate (DEHP), dibutylphthalate (DBP), diisododecylphthalate (DIDP) and 4-nonylphenol (4-NP) were oestrogenic in the yeast estrogen screen (YES) assay and potently oestrogenic in the MVLN and E-SCREEN assays at environmentally relevant concentrations. DINP and 4-NP were mutagenic in the Ames assay and also induced significant levels of unscheduled DNA synthesis and DNA strand breakage. Significant induction in the percentage of cells containing micronuclei was observed after treatment with DINP, DEHP and 4-NP. In addition, sewage effluents from sewage treatment plants (STPs) in the Border, Midlands and Western (BMW) region of Ireland were significantly oestrogenic in the YES assay. Moreover, analysis of levels of phthalates and alkylphenol identified in Irish rivers receiving treated effluent showed potent oestrogenicity in the YES assay. The proliferative and genotoxic ability of the phthalates and alkylphenol, and the oestrogenicity of the treated effluents reported here, is significant as these EDCs and EDCs within the effluent may play a role in the etiology of human abnormalities.

Share and Cite:

K. Quinn-Hosey, J. Roche, A. Fogarty and C. Brougham, "A Toxicological Assessment of Endocrine Disrupting Chemicals Found in the BMW (Border, Midland and Western) Region of Ireland," Journal of Environmental Protection, Vol. 3 No. 4, 2012, pp. 304-315. doi: 10.4236/jep.2012.34039.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] S. Jobling, T. Reynolds, R. White, M. G. Parker and J. P. Sumpter, “A Variety of Environmentally Persistent Chemi- cals, Including Some Phthalate Plasticisers Are Weakly Estrogenic,” Environmental Health Perspectives, Vol. 103, No. 6, 1995, pp. 582-587. doi:10.1289/ehp.95103582
[2] C. E. Purdom, P. A. Hardiman, V. J. Bye, N. C. Eno, C. R. Tyler and J. P. Sumpter, “Estrogenic Effects of Effluents from Sewage Treatment Works,” Chemistry and Ecology, Vol. 8, No. 4, 1994, pp. 275-285. doi:10.1080/02757549408038554
[3] K. L. Thorpe, G. Maack, R. Benstead and C. R. Tyler, “Estrogenic Wastewater Treatment Works Effluents Re- duce Egg Production in Fish,” Environmental Science and Technology, Vol. 43, No. 8, 2009, pp. 2976-2982. doi:10.1021/es803103c
[4] K. E. Liney, S. Jobling, J. A. Shears, P. Simpson and C. R. Tyler, “Assessing the Sensitivity of Different Life Stages for Sexual Disruption in Roach (Rutilus rutilus) Exposed to Effluents from Wastewater Treatment Works,” Envi- ronmental Health Perspectives, Vol. 113, No. 10, 2005, pp. 1299-1307. doi:10.1289/ehp.7921
[5] M. Y. Gross-Sorokin, S. D. Roast and G. C. Brighty, “Assessment of Feminisation of Male Fish in English Riv- ers by the Environemnt Agency of England and Wales,” Environmental Health Perspectives, Vol. 114, No. 1, 2006, pp. 147-151. doi:10.1289/ehp.8068
[6] S. Markman, I. A. Guschina, S. Barnsley, K. L. Buchanan, D. Pascoe and C. T. Muller, “Endocrine Disrupting Chemi- cals Accumulate in Earthworms Exposed to Sewage Ef- fluent,” Chemosphere, Vol. 70, No. 1, 2007, pp. 119-125. doi:10.1016/j.chemosphere.2007.06.045
[7] E. J. Routledge and J. P. Sumpter, “Estrogenic Activity of Surfactants and some of Their Degradation Products As- sessed Using a Recombinant Yeast Screen,” Environ- mental Toxicology and Chemistry, Vol. 15, No. 3, 1996, pp. 241-248. doi:10.1002/etc.5620150303
[8] M. Pons, D. Gagne, J. C. Nicolas and M. Mehtali, “A New Cellular Model of Response to Estrogens: A Biolu- minescent Test to Characterise (Anti) Estrogen Mole- cules,” Biotechniques, Vol. 9, No. 4, 1990, pp. 450-459.
[9] A. M. Soto, K. L. Chung and C. Sonnenschein, “The Pesti- cides Endosulfan, Toxaphene, and Dieldrin Have Estro- genic Effects on Human Estrogen-Sensitive Cells,” Envi- ronmental Health Perspectives, Vol. 102, No. 4, 1994, pp. 380-383. doi:10.1289/ehp.94102380
[10] A. M. Soto, T. M. Lin, H. Justicia, R. M. Silvia and C. Sonnenschein, “An “in Culture” Bioassay to Assess the Estrogenicity of Xenobiotics (E-SCREEN),” In: T. Colburn and C. Clement, Eds., Chemically induced Alterations in Sexual and Functional Development: The Wildlife/Hu- man Connection, Princeton Scientific Publishing, Prince- ton, 1992, pp. 295-309.
[11] N. P. Singh, M. T. McCoy, R. R. Tice and E. L. Schnei- der, “A Simple Technique for Quantitation of Low Levels of DNA Damage in Individual Cells,” Experimental Cell Research, Vol. 175, No. 1, 1988, pp. 184-191. doi:10.1016/0014-4827(88)90265-0
[12] F. Naji-Ali, B. M. Hasspieler, D. Haffner and K. Adeli, “Human Bioassays to Assess Environmental Genotoxicity: Development of a DNA Repair Assay in HepG2 Cells,” Clinical Biochemistry, Vol. 27, No. 6, 1994, pp. 441-448. doi:10.1016/0009-9120(94)00050-6
[13] M. Fenech, “The Cytokinesis-Block Micronucleus Tech- nique: A Detailed Description of the Method and Its Ap- plication to Genotoxicity Studies in Human Populations,” Mutation Research, Vol. 285, No. 1, 1993, pp. 35-44. doi:10.1016/0027-5107(93)90049-L
[14] B. N. Ames, J. McCann and E. Yamaski, “Methods for Detecting Carcinogens and Mutagens with the Salmo- nella/Mammalian-Microsome Mutagenicity Test,” Muta- tion Research, Vol. 31, No. 1, 1975, pp. 347-364.
[15] B. N. Ames, W. E. Durston, E. Yamasaki and F. D. Lee, “Carcinogens Are Mutagens: A Simple Test System Com- bining Liver Homogenates for Activation and Bacteria for Detection,” Proceedings of the National Academy of Sci- ences of USA, Vol. 70, No. 8, 1973, pp. 2281-2285. doi:10.1073/pnas.70.8.2281
[16] D. M. Maron and B. N. Ames, “Revised Methods for the Salmonella Mutagenicity Test,” Mutation Research, Vol. 113, No. 3-4, 1983, pp. 173-215. doi:10.1016/0165-1161(83)90010-9
[17] P. Gee, D. M. Maron and B. N. Ames, “Detection and Classification of Mutagens: A Set of Base-Specific Sal- monella Tester Strains,” Proceedings of the National Academy of Sciences of USA, Vol. 91, No. 24, 1994, pp. 11606-11610. doi:10.1073/pnas.91.24.11606
[18] A. M. Reid, C. A. Brougham, A. M. Fogarty and J. J. Roche, “Isocratic LC Methods for the Trace Analysis of Phthalates and 4-Nonylphenol in Varying Types of Land- fill and Adjacent Run-Offs,” Toxicological and Environ- mental Chemistry, Vol. 89, No. 3, 2007, pp. 399-410. doi:10.1080/02772240601116613
[19] A. M. Reid, C. A. Brougham, A. M. Fogarty and J. J. Roche, “Accelerated Solvent-Based Extraction and En- richment of Selected Plasticisers and 4-Nonylphenol, and Extraction of Tin from Organotin Sources in Sediments, Sludges and Leachate Soils,” Analytica Chimica Acta, Vol. 634, No. 2, 2008, pp. 197-204. doi:10.1016/j.aca.2008.12.032
[20] A. M. Soto, C. Sonnenschein, K. L. Chung, M. F. Fer- nandez, N. Olea and F. Serrano, “The E-SCREEN Assay as a Tool to Identify Estrogens: An Update on Estrogenic Environmental Pollutants,” Environmental Health Per- spectives, Vol. 103, No. 7, 1995, pp. 113-122.
[21] C. R. Kent, J. J. Eady, G. M. Ross and G. G. Steel, “The Comet Moment as a Measure of DNA Damage in the Comet Assay,” International Journal of Radiation Biol- ogy, Vol. 67, No. 6, 1995, pp. 655-660.
[22] H. Dechaud, C. Ravard, F. Claustrat, A. B. de la Pierriere and M. Pugeat, “Xenestrogen Interaction with Human Sex Hormone-Binding Globulin (hSHBG),” Steroids, Vol. 64, No. 5, 1999, pp. 328-334. doi:10.1016/S0039-128X(98)00114-7
[23] C. F. Wilkinson and J. C. Lamb IV, “The Potential Health Effects of Phthalate Esters in Children’s Toys: A Review and Risk Assessment,” Regulatory Toxicology and Phar- macology, Vol. 30, No. 2, 1999, pp. 140-155. doi:10.1006/rtph.1999.1338
[24] S. J. Waterman, J. L. Ambroso, L. H. Keller, G. W. Trim- mer, A. I. Nikiforov and S. B. Harris, “Developmental Toxicity of Di-Isodecyl and Di-Isononyl Phthalates in Rats,” Reproductive Toxicology, Vol. 13, No. 2, 1999, pp. 131-136. doi:10.1016/S0890-6238(99)00002-7
[25] R. Kavlock, K. Boekelheide, R. Chapin, M. Cunningham, E. Faustman, P. Foster, M. Golub, R. Henderson, I. Hin- berg, R. Little, J. Seed, K. Shea, S. Tabacova, R. Tyl, P. Williams, T. Zacharewski, M. Shelby, C. Portier, G. Jah- nke, L. Goldman, J. Moore, A. Iannucci and A. Walker, “NTP Centre for the Evaluation of Risks to Human Re- production: Phthalates Expert Panel Report on the Re- productive and Developmental Toxicity of Di-Isononyl Phthalate,” Reproductive Toxicology, Vol. 16, No. 5, 2000, pp. 679-708.
[26] M. A. Kelly, A. M. Reid, K. M. Quinn-Hosey, A. M. Fogarty, J. J. Roche and C. A. Brougham, “Investigation of the Estrogenic Risk to Feral Male Brown Trout (Salmo trutta) in the Shannon International River Basin District of Ireland,” Ecotoxicology and Environemntal Safety, Vol. 73, No. 7, 2010, pp. 1658-1665. doi:10.1016/j.ecoenv.2010.08.018
[27] L. A. Haighton, J. J. Hlywka, J. Doull, R. Kroes, B. S. Lynch and I. C. Munro, “An Evaluation of the Possible Carcinogenicity of Bisphenol A to Humans,” Regulatory Toxicology and Pharmamcology, Vol. 35, No. 2, 2002, pp. 238-254. doi:10.1006/rtph.2001.1525
[28] S. Cotelle and J. F. Ferard, “Comet Assay in Genetic Ecotoxicology: A Review,” Environmental and Molecu- lar Mutagenesis, Vol. 34, No. 4, 1999, pp. 246-255. doi:10.1002/(SICI)1098-2280(1999)34:4<246::AID-EM4>3.0.CO;2-V
[29] D. Anderson, M. M. Dobrzynska and N. Basaran, “Effect of Various Genotoxins and Reproductive Toxins in Hu- man Lymphocytes and Sperm in the Comet Assay,” Tera- togenesis, Carcinogenesis, and Mutagenesis, Vol. 17, No. 1, 1997, pp. 29-43. doi:10.1002/(SICI)1520-6866(1997)17:1<29::AID-TCM5>3.0.CO;2-H
[30] C. N. Martin, A. C. McDermid and R. C. Garner, “Test- ing of Known Carcinogens and Noncarcinogens for Their Ability to Induce Unscheduled DNA aynthesis in HeLa Cells,” Cancer Research, Vol. 38, No. 8, 1978, pp. 2621- 2627.
[31] K. Al-Sabti and C. D. Metcalfe, “Fish Micronuclei for Assessing Genotoxicity in Water,” Mutation Research, Vol. 343, No. 2-3, 1995, pp. 121-135. doi:10.1016/0165-1218(95)90078-0
[32] M. Fenech and A. A. Morley, “Kinetochore Detection in Micronuclei: An Alternative Method for Measuring Chro- mosome Loss,” Mutagenesis, Vol. 4, No. 2, 1989, pp. 98- 104. doi:10.1093/mutage/4.2.98
[33] J. D. Tucker and R. J. Preston, “Chromosome Aberrations, Micronuclei, Aneuploidy, Sister Chromatid Exchanges, and Cancer Risk Assessment,” Mutation Research, Vol. 365, No. 1-3, 1996, pp. 147-159.
[34] H. Tarrant, N. Llewellyn, A. Lyons, N. Tattersall, S. Wylde, G. Mouzakitis, M. Maloney and C. McKenzie, “Endocrine Disruptors in the Irish Aquatic Environment. Environmental RTDI Programme 2000-2006,” Johnstown Castle, Co. Wexford, 2005.
[35] C. Desbrow, E. J. Routledge, G. C. Brighty, J. P. Sumpter and M. Waldock, “Identification of Estrogenic Chemicals in STW Effluent. 1. Chemical Fractionation and in Vitro Biological Screening,” Environmental Science and Tech- nology, Vol. 32, No. 11, 1998, pp. 1549-1558. doi:10.1021/es9707973
[36] S. Jobling, R. Williams, A. Johnson, A. Taylor, M. Gross- Sorokin, M. Nolan, C. R. Tyler, R. van Aerle, E. Santos and G. Brighty, “Predicted Exposures to Steroid Estro- gens in U.K. Rivers Correlate With Widespread Sexual Disruption in Wild Fish Populations,” Environmental Health Perspectives, Vol. 114, No. 1, 2006, pp. 32-39. doi:10.1289/ehp.8050
[37] A. C. Belfroid, A. Van der Horst, A. D. Vethaak, A. J. Schafer, G. B. J. Rijs, J. Wegener and W. P. Cofino, “Analysis and Occurrence of Estrogenic Hormones and Their Glucuronides in Surface Water and Waste Water in The Netherlands,” The Science of the Total Environment, Vol. 225, No. 1-2, 1999, pp. 101-108. doi:10.1016/S0048-9697(98)00336-2
[38] P. Spengler, W. Korner and J. W. Metzger, “Substances with Estrogenic Activity in Effluents of Sewage Treat- ment Plants in Southwestern Germany. 1. Chemical Analy- sis,” Environmental Toxicology and Chemistry, Vol. 20, No. 10, 2001, pp. 2133-2141. doi:10.1002/etc.5620201001
[39] W. Korner, P. Spengler, U. Bolz, W. Schuller, V. Hanf and J. W. Metzger, “Substances with Estrogenic Activity in Effluents of Sewage Treatment Plants in Southwestern Germany. 2. Biological Analysis,” Environmental Toxi- cology and Chemistry, Vol. 20, No. 10, 2001, pp. 2142- 2151. doi:10.1002/etc.5620201002
[40] R. A. Rudel, S. J. Melly, P. W. Geno, G. Sun and J. G. Brody, “Identification of Alkylphenols and other Phenolic Compounds in Wastewater, Septage, and Groundwater on Cape Cod, Massachusetts,” Environmental Science Tech- nology, Vol. 32, No. 7, 1998, pp. 861-869. doi:10.1021/es970723r
[41] J. H. Shen, B. Gutendorf, H. H. Vahl, L. Shen and J. Westendorf, “Toxicological Profile of Pollutants in Sur- face Water from an Area in Taihu Lake, Yangtze Delta,” Toxicology, Vol. 166, No. 1-2, 2001, pp. 71-78. doi:10.1016/S0300-483X(01)00439-5
[42] J. H. Hodgert, T. R. Zacharewski and G. L. Hammond, “Interactions between Human Plasma Sex Hormone-Bind- ing Globulin and Xenobiotic Ligands,” The Journal of Steroid Biochemistry and Molecular Biology, Vol. 75, No. 2-3, 2000, pp. 167-176. doi:10.1016/S0960-0760(00)00168-0
[43] A. O. Ifelebuegu, J. N. Lester, J. Churchley and E. Cart- mell, “Removal of an Endocrine Disrupting Chemical (17 Alpha-Ethinylestradiol) from Wastewater Effluent by Ac- tivated Carbon Adsorption: Effects of Activated Carbon Type and Competitive Adsorption,” Environmental Tech- nology, Vol. 27, No. 12, 2006, pp. 1343-1349. doi:10.1080/09593332708618748
[44] N. Jonkers, H. P. Kohler, A. Dammshauser and W. Giger, “Mass Flows of Endocrine Disruptors in the Glatt River during Varying Weather Conditions,” Environmental Pol- lution, Vol. 157, No. 3, 2009, pp. 714-723. doi:10.1016/j.envpol.2008.11.029
[45] P. Matthiessen, D. Arnold, A. C. Johnson, T. J. Pepper, T. G. Pottinger and K. G. Pulman, “Contamination of Head- water Streams in the United Kingdom by Oestrogenic Hormones from Livestock Farms,” The Science of the Total Environment, Vol. 367, No. 2-3, 2006, pp. 616-630. doi:10.1016/j.scitotenv.2006.02.007
[46] A. C. Johnson, R. J. Williams and P. Matthiessen, “The Potential Steroid Hormone Contribution of Farm Animals to Freshwaters, the United Kingdom as a Case Study,” The Science of the Total Environment, Vol. 362, No. 1-3, 2006, pp. 166-178. doi:10.1016/j.scitotenv.2005.06.014
[47] W. Korner, U. Bolz, W. Submuth, G. Hiller, W. Schuller, V. Hanf and H. Hagenmaier, “Input/Output Balance of Estrogenic Active Compounds in a Major Municipal Sew- age Plant in Germany,” Chemosphere, Vol. 40, No. 9-11, 2000, pp. 1131-1142. doi:10.1016/S0045-6535(99)00362-8
[48] G. H. Panter, R. S. Thompson, N. Beresford and J. P. Sumpter, “Transformation of a Non-Estrogenic Steroid Metabolite to an Estrogenically Active Substance by Mini- mal Bacterial Activity,” Chemosphere, Vol. 38, No. 15, 1999, pp. 3579-3596. doi:10.1016/S0045-6535(98)00572-4
[49] T. A. Ternes, P. Kreckel and J. Mueller, “Behaviour and Occurrence of Estrogens in Municipal Sewage Treatment Plants—II. Aerobic Batch Experiments with Activated Sludge,” The Science of the Total Environment, Vol. 225 No. 1-2, 1999, pp. 91-99. doi:10.1016/S0048-9697(98)00335-0
[50] D. L. Davis and H. L. Bradlow “Can Environmental Es- trogens Cause Breast Cancer?” Scientific American Oc- tober, 1995, pp. 144-149.
[51] T. J. oppari, J. C. Larsen, P. Christiansen, A. Giwercman, P. Grandjean, L. J. Guilette, B. Jegou, T. K. Jensen, P. Jouannet, N. Keiding, H. Leffers, J. A. McLachlan, O. Meyer, J. Muller, E. R.-D. Meyts, T. Scheike, R. Sharpe, J. Sumpter and N. E. Skakkebaek, “Male Reproductive Health and Environmental Xenestrogens,” Environmental Health Perspectives, Vol. 104, No. 4, 1996, pp. 741-803.
[52] R. M. Sharpe and N. E. Skakkebaek, “Are Oestrogens Involved in Falling Sperm Counts and Disorders of the Male Reproductive Tract?” The Lancet, Vol. 341, No. 8857, 1993, pp. 1392-1395. doi:10.1016/0140-6736(93)90953-E
[53] E. Carlsen, A. Giwercman, N. Keiding and N. E. Skak- kebaek, “Declining Semen Quality and Increasing Inci- dence of Testicular Cancer: Is There a Common Cause?” Environmental Health Perspectives, Vol. 103, No. 7, 1995, pp. 137-139.
[54] S. F. Arnold, D. M. Klotz, B. M. Collins, P. M. Vonier, L. J. Guillette and J. A. McLachlan, “Synergistic Activation of Estrogen Receptor with Combinations of Environmental Chemicals,” Science, Vol. 272, No. 5267, 1996, pp. 1489- 1492. doi:10.1126/science.272.5267.1489
[55] S. M. Dempsey and M. J. Costello, “A Review of Estro- gen Mimicking Chemicals in Relation to Water Quality in Ireland,” Johnstown Castle, Co. Wexford, 1998.

Copyright © 2022 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.