Studies of Uptake, Elimination, and Late Effects in Atlantic Salmon (Salmo salar) Dietary Exposed to Di-2-Ethylhexyl Phthalate (DEHP) During Early Life

  • A. NormanEmail author
  • H. Börjeson
  • F. David
  • B. Tienpont
  • L. Norrgren


The phthalate esters are a group of industrial chemicals considered to have endocrine-disrupting properties. The most common tonnage product among these, di-2-ethylhexyl phthalate (DEHP), is widely spread in the environment. The objectives with the present work were to study uptake and metabolism of orally administered DEHP and its major metabolite mono-2-ethyl hexyl phthalate (MEHP) and to evaluate the impact of early life exposure on sex differentiation in Atlantic salmon. The feeding with contaminated diet started immediately after yolk sac resorption and continued for 4 weeks. Nominal concentrations of DEHP in the diet were 400 (measured 359), 800 (measured 827), and 1500 (measured 1648) mg DEHP/kg and a control group was fed food mixed with solvent. After the exposure period, fish were fed non-contaminated diet until final sampling 4 months post-exposure. There were no effects on growth or survival of the fish and no late effects on hepatosomatic index or sex ratio. However, the histological examination of gonads from fish exposed to 1500 mg DEHP/kg revealed a small but significant incidence (3%) of intersex fish (ovo-testis). Chemical residues of DEHP and MEHP were analyzed weekly during the first 3 months of the post-exposure period. Both DEHP and MEHP were rapidly eliminated to near background levels within one week post exposure. The study indicates that exposure of Atlantic salmon to relatively high concentrations of DEHP during a sensitive part of the life cycle may interfere with gonad differentiation.


Phthalate Atlantic Salmon DEHP Nominal Concentration Phthalate Ester 
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This work was financially supported by the European Council for Plasticizers and Intermediates (ECPI) and by grants from the ReproSafe-program at the Swedish environmental Protection Agency. The Swedish Board of Fisheries, Älvkarleby is acknowledged for keeping the fish and for support during sampling.


  1. Andersen Ø, Eijsink VGH, Thomassen M (2000) Multiple variants of the peroxisome proliferator-activated receptor (PPAR) γ are expressed in the liver of Atlantic salmon (Salmo salar). Gene 255:411–418CrossRefGoogle Scholar
  2. Balafas D, Shaw KJ, Whitfield FB (1999) Phthalate and adipate esters in Australian packaging materials. Food Chem 65:279–287CrossRefGoogle Scholar
  3. Barron MG, Tarr BD, Hayton WL (1987) Temperature dependence of di-2-ethylhexyl phthalate (DEHP) pharmacokinetics in rainbow trout. Toxicol Appl Pharmacol 88:305–312CrossRefGoogle Scholar
  4. Barron MG, Schultz IR, Hayton WL (1989) Presystemic branchial metabolism limits di-2-ethylhexyl phthalate accumulation in Fish. Toxicol Appl Pharmacol 98:49–57CrossRefGoogle Scholar
  5. Barron MG, Albro PW, Hayton WL (1995) Biotransformation of di(2-ethylhexyl) phthalate by rainbow trout. Environ Toxicol Chem 14(5):873–876Google Scholar
  6. Blom A, Ekman E, Johannisson A, Norrgren L, Pesonen M (1998) Effects of xenoestrogenic environmental pollutants on the proliferation of human breast cancer cell line (MCF-7). Arch Environ Contam Toxicol 34:306–310CrossRefGoogle Scholar
  7. David F, Sandra P (2001) Phthalate esters in the environment. Monitoring program for the determination of phthalates in air, vegetation, cattle feed, milk and fish in the Netherlands (1999–2001). Report No. ECPI-2001-10. Research Institute of Chromatography, Kortrijk, BelgiumGoogle Scholar
  8. Davis BJ, Maronpot RR, Heindel JJ (1994) Di-(2-ethylhexyl) phthalate suppresses estradiol and ovulation in cycling rats. Toxicol Appl Pharmacol 128:216–223CrossRefGoogle Scholar
  9. Devlin RH, Nagahama Y (2002) Sex determination and sex differentiation in fish: an overview of genetic, physiological, and environmental influences. Aquaculture 208:191–364CrossRefGoogle Scholar
  10. Earls AO, Axford IP, Braybrook JH (2003) Gas chromatography-mass spectrometry determination of the migration of phthalate plasticisers from polyvinyl chloride toys and childcare articles. J Chromatogr 983A:237–246CrossRefGoogle Scholar
  11. Giam CS, Chan HS, Neff GS (1975) Sensitive methods for determination of phthalate ester plasticizers in open-ocean biota samples. Anal Chem 47(13):2225–2229CrossRefGoogle Scholar
  12. Goetz FW, Donaldson EM, Hunter GA, Dye HM (1979) Effects of estradiol-17β and 17α-methyl testosterone on gonadal differentiation in the Coho salmon, Oncorhynchus kisutch. Aquaculture 17:267–278CrossRefGoogle Scholar
  13. Gray LE Jr, Wolf C, Lambright C, Mann P, Price M, Cooper RL, Ostby J (1999) Administration of potentially antiandrogenic pesticides (procymidone, linuron, iprodione, chlozolinate, p,p’-DDE, and ketoconazole) and toxic substances (dibutyl- and diethylhexyl phthalate, PCB 169, and ethane dimethane sulphonate) during sexual differentiation produces diverse profiles of reproductive malformations in the male rat. Toxicol Ind Health 15:94–118CrossRefGoogle Scholar
  14. Harris CA, Henttu P, Parker MG, Sumpter JP (1997) The estrogenic activity of phthalate esters in vitro. Environ Health Perspect 105(8):802–811Google Scholar
  15. Haux C, Norberg B (1985) The influence of estradiol-17β on the liver content of protein, lipids, glycogen and nucleic acids in juvenile rainbow trout, Salmo gairdnerii. Comp Biochem Physiol 81B(2):275–279Google Scholar
  16. Isenberg JS, Kamendulis LM, Ackley DC, Smith JH, Pugh G Jr, Lington AW, McKee RH, Klaunig JE (2001) Reversibility and persistence of di-2-ethylhexyl phthalate (DEHP)- ands Phenobarbital-induced hepatocellular changes in rodents. Toxicol Sci 64:192–199CrossRefGoogle Scholar
  17. Jobling S, Nolan M, Tyler CR, Brighty G, Sumpter JP (1998) Widespread sexual disruption in wild fish. Environ Sci Technol 32:2498–2506CrossRefGoogle Scholar
  18. Johnstone R, Simpson TH, Youngson AF (1978) Sex reversal in salmonid culture. Aquaculture 13:115–134CrossRefGoogle Scholar
  19. Kim E-J, Kim J-W, Lee S-K (2002) Inhibition of oocyte development in Japanese medaka (Oryzias latipes) exposed to di-2-ethylhexyl phthalate. Environ Int 28:359–365CrossRefGoogle Scholar
  20. Koger CS, Teh SJ, Hinton DE (2000) Determining the sensitive developmental stages of intersex induction in medaka (Oryzias latipes) exposed to17β-estradiol or testosterone. Mar Environ Res 50:201–206CrossRefGoogle Scholar
  21. Larsson DGJ, Adolfsson-Erici M, Parkkonen J, Petterson M, Berg AH, Olsson P-E, Förlin L (1999) Ethinyloestradiol: an undesired fish contraceptive? Aquat Toxicol 45:91–97CrossRefGoogle Scholar
  22. Lapinskas PJ, Brown S, Leesnitzer LM, Blanchard s, Swanson C, Cattley RC, Corton JC (2005) Role of PPARα in mediating the effects of phthalates and metabolites in the liver. Toxicology 207:149–163CrossRefGoogle Scholar
  23. Lovekamp TN, Davis BJ (2001) Mono-(2-ethylhexyl) phthalate suppresses aromatase transcript levels and estradiol production in cultured rat granulosa cells. Toxicol Appl Pharmacol 172:217–224CrossRefGoogle Scholar
  24. Matthiessen P, Allen Y, Bamber S, Craft J, Hurst M, Hutchinson T, Feist S, Katsiadaki I, Kirby M, Robinson C, Scott S, Thain J, Thomas K (2002) The impact of oestrogenic and androgenic contamination on marine organisms in the United Kingdom: summary of the EDMAR programme. Mar Environ Res 54:645–649CrossRefGoogle Scholar
  25. Metcalfe CD, Metcalfe TL, Kiparissis Y, Koenig BG, Khan C, Hughes RJ, Croley TR, March RE, Potter T (2001) Estrogenic potency of chemicals detected in sewage treatment plant effluents as determined by in vivo assays with Japanese medaka (Oryzias latipes). Environ Toxicol Chem 20(2):297–308CrossRefGoogle Scholar
  26. Moody DE, Reddy JK (1978) Hepatic peroxisome (Microbody) proliferation in rats fed plasticizers and related compounds. Toxicol Appl Pharmacol 45:497–504CrossRefGoogle Scholar
  27. Mu Y-M, Yanase T, Nishi Y, Waseda N, Oda T, Tanaka A, Takayanagi R, Nawata H (2000) Insulin sensitizer, troglitazone, directly inhibits aromatase activity in human ovarian granulosa cells. Biochem Biophys Res Commun 271:710–713CrossRefGoogle Scholar
  28. Norrgren L, Blom A, Andersson PL, Börjeson H, Larsson DGJ, Olsson P-E (1999) Effects of potential xenoestrogens (DEHP, nonylphenol and PCB) on sexual differentiation in juvenile Atlantic salmon (Salmo salar). Aquat Ecosyst Health Manage 2:311–317CrossRefGoogle Scholar
  29. Ortiz-Zarragoitia M, Cajaraville MP (2000) Environmental estrogenic compounds alter peroxisomal functions in fish. Comp Biochem Physiol A 126:S1–S113Google Scholar
  30. Page BD, Lacroix GM (1995) The occurrence of phthalate ester and di-2-ethylhehyl adipate plasticizers in Canadian packaging and food sampled in 1985-1989: a survey. Food Addit Contam 12(1):129–151Google Scholar
  31. Pandian TJ, Sheela SG (1995) Hormonal induction of sex reversal in fish. Aquaculture 138:1–22CrossRefGoogle Scholar
  32. Parks LG, Ostby JS, Lambright CR, Abbott B, Klinefelter GR, Barlow NJ, Gray LE Jr (2000) The plasticizer diethylhexyl phthalate induces malformation by decreasing fetal testosterone synthesis during sexual differentiation in the male rat. Toxicol Sci 58:339–349CrossRefGoogle Scholar
  33. Reddy JK, Lalwani ND (1983) Carcinogenesis by hepatic peroxisome proliferators: evaluation of the risk of hypolipidemic drugs and industrial plasticizers to humans. CRC Critic Rev Toxicol 12:1–58Google Scholar
  34. Ruyter B, Andersen Ø, Dehli A, Östlund Farrants A-K, Gjøen T, Thomassen MS (1997) Peroxisome proliferator activated receptors in Atlantic salmon (Salmo salar): effects on PPAR transcription and acyl-CoA oxidase activity in hepatocytes by peroxisome proliferators and fatty acids. Biochim Biophys Acta 1348:331–338Google Scholar
  35. Scarano LJ, Calabrese EJ, Kostecki PT, Baldwin LA, Leonard DA (1994) Evaluation of a rodent peroxisome proliferator in two species of freshwater fish: Rainbow trout (Onchorynchus mykiss) and Japanese Medaka (Oryzias latipes). Ecotox Environ Saf 29:13–19CrossRefGoogle Scholar
  36. Soto AM, Sonnenschein C, Chung KL, Fernandez MF, Olea N, Olea Serrano F (1995) The E-screen assay as a tool to identify estrogens; an update on estrogenic environmental pollutants. Environ Health Perspect 103(7):113–122Google Scholar
  37. Sower SA, Dickhoff WW, Flagg TA, Michall JL, Mahnken CVW (1984) Effects of estradiol and diethylstilbestrol on sex reversal and mortality in Atlantic salmon (Salmon salar). Aquaculture 43:75–81CrossRefGoogle Scholar
  38. Staples CA, Peterson DR, Parkerton TF, Adams WJ (1997) The environmental fate of phthalate esters: A literature review. Chemosphere 35(4):667–749CrossRefGoogle Scholar
  39. Tan GH (1995) Residue levels of phthalate esters in water and sediment samples from the Klang river basin. Bull Environ Contam Toxicol 54:171–176CrossRefGoogle Scholar
  40. Thomann RV (1989) Bioaccumulation model of organic chemical distribution in aquatic food chains. Environ Sci Technol 23:699–707CrossRefGoogle Scholar
  41. Tienpont B, David F, Sandra P (in prep.). In preparation for J ChromatogrGoogle Scholar
  42. Vethaak AD, Rijs GBJ, Schrap SM, Ruiter H, Gerristen A, Lahr J (2002) Estrogens and xeno-estrogens in the aquatic environment of the Netherlands, occurrence, potency and biological effects, RIZA/RIKZ Report no 2002.001, Dictorate-General for Public Works and Waste ManagementGoogle Scholar
  43. Wahl HG, Hoffman A, Häring H-U, Leibich HM (1999) Identification of plasticizers in medical products by a combined direct thermodesorption-cooled injection system and gas chromatography-mass spectrometry. J Chromatogr 847A:1–7CrossRefGoogle Scholar
  44. Zacharewski TR, Meek MD, Clemons JH, Wu ZF, Fielden MR, Matthews JB (1998) Examination of the in vitro and in vivo estrogenic activities of eight commercial phthalate esters. Toxicol Sci 46:282–293CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • A. Norman
    • 1
    • 4
    Email author
  • H. Börjeson
    • 2
  • F. David
    • 3
  • B. Tienpont
    • 3
  • L. Norrgren
    • 1
    • 4
  1. 1.Department of Biomedical Sciences and Veterinary Public HealthSwedish University of Agricultural SciencesUppsalaSweden
  2. 2.Swedish Fish Health Control ProgramÄlvkarlebySweden
  3. 3.Research Institute for ChromatographyKortvijkBelgium
  4. 4.Centre for Reproductive BiologyUppsalaSweden

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