Environmental Science and Pollution Research

, Volume 20, Issue 2, pp 723–737 | Cite as

Histopathological lesions and DNA adducts in the liver of European flounder (Platichthys flesus) collected in the Seine estuary versus two reference estuarine systems on the French Atlantic coast

  • Jérôme CachotEmail author
  • Yan Cherel
  • Thibaut Larcher
  • Annie Pfohl-Leszkowicz
  • Jean Laroche
  • Louis Quiniou
  • Jocelyne Morin
  • Julien Schmitz
  • Thierry Burgeot
  • Didier Pottier
Ecotoxicology of estuaries in France and Québec, Canada


An epidemiological survey was conducted in the Seine estuary and in two smaller and relatively preserved estuaries on the French Atlantic coast in order to estimate the occurrence of liver lesions in European flounder, Platichthys flesus, and also to seek putative risk factors for the recorded pathologies. Four hundred and seventy-eight fish of both sexes and of different size ranges were sampled in the three studied areas, 338 of which in the Seine estuary. All fish were examined for histopathological liver lesions, while DNA adducts and otoliths were analyzed on a subsample. Five categories of hepatic lesions were recorded with the following prevalence for the Seine estuary: 36.7 % inflammations, 8 % parasites (mainly encysted nematodes), 6.5 % foci of cellular alteration (FCA), 5.3 % foci of necrosis or regeneration (FNR), and 1.5 % tumors. Inflammation occurrence increased according to age, contrary to parasitic infestations and FCA which were more prevalent in young fish, notably those of <1 year old (group 0). Tumors were only observed in females of more than two winters. Females exhibited a higher prevalence of tumors (3.0 %) and FCA (6.5 %) than males (0 and 2.6 %, respectively). Parasitic and infectious lesions and FNR were equally distributed in males and females. The prevalence of FNR was also shown to vary according to sampling season, with significantly more occurrences of liver necrosis in the fish collected in summer than in spring. Spatial differences were observed with a higher occurrence of encysted parasites in flounders from the upper Seine estuary, while inflammations predominated in flounders living downstream. Temporal trends were also noted, with an increased prevalence of parasitic infestations, inflammations, and FCA in the 2002–2003 period in comparison to the 1996–1997 one. The three flounder populations from the Seine estuary (Normandy), Ster estuary (Brittany), and Bay of Veys (Normandy) showed different spectra of hepatic lesions. Flounders from the Bay of Veys had relatively few liver lesions as compared to flounders from the two other estuaries. Flounders from the Ster estuary exhibited the highest prevalence of parasites (37.2 %) and inflammations (51.1 %). Finally, FCA and liver tumors occurred at very similar levels in both flounder populations from the Seine and the Ster estuaries. Group 0 flounders inhabiting the upper Seine estuary were more prone to parasitic and pre-neoplastic hepatic lesions and had higher levels of liver DNA adducts than the older ones living downstream. It was postulated that group 0 European flounders may serve as valuable bioindicators for assessing the quality of estuarine waters and the health status of euryhaline fish populations.


Epidemiological study European flounder Seine estuary Liver histopathology Necrosis FCA Tumors Inflammations Parasites DNA adducts Pollution 



Oslo–Paris Convention on the Protection of the Marine Environment of the North-East Atlantic


Diagonal radioactive zone


European College of Veterinary Pathologists



This study was supported by the Agence de l’Eau Seine-Normandie, the Seine-Aval program, and Ifremer. The authors would like to thank the Cellule du Suivi du Littoral Haut Normand (CSLHN, Le Havre) and the crews from the oceanographic vessels Gwen Drez (Ifremer) and Esturial (Irstea) for their technical assistance in fish collection.


  1. Aas E, Beyer J, Jonsson G, Reichert WL, Andersen OK (2001) Evidence of uptake, biotransformation and DNA binding of polyaromatic hydrocarbons in Atlantic cod and corkwing wrasse caught in the vicinity of an aluminium works. Mar Environ Res 52:213–229CrossRefGoogle Scholar
  2. Amara R, Selleslagh J, Billon G, Minier C (2009) Growth and condition of 0-group European flounder, Platichthys flesus as indicator of estuarine habitat quality. Hydrobiologia 627:87–98CrossRefGoogle Scholar
  3. Beck MW, Heck KL Jr, Able KW, Childers DL, Eggleston DB, Gillanders BM, Halpern B, Hays CG, Hoshino K, Minello TJ, Orth RJ, Sheridan PF, Weinstein MP (2001) The identification, conservation, and management of estuarine and marine nurseries for fish and invertebrates. BioScience 51:633–641CrossRefGoogle Scholar
  4. Benejam L, Benito J, García-Berthou E (2010) Decreases in condition and fecundity of freshwater fishes in a highly polluted reservoir. Water Air Soil Pollut 210:231–242CrossRefGoogle Scholar
  5. Bogovski S, Lang T, Mellergaard S (1999) Histopathological examinations of liver nodules in flounder (Platichthys flesus L.) from the Baltic Sea. ICES J Mar Sci 56:148–151CrossRefGoogle Scholar
  6. Brooks ML, Fleishman E, Brown LR, Lehman PW, Werner I, Scholz N, Mitchelmore C, Lovvorn JR, Johnson ML, Schlenk D, van Drunick S, Drever JI, Stoms DM, Parker AE, Dugdale R (2012) Life histories, salinity zones, and sublethal contributions of contaminants to pelagic fish declines illustrated with a case study of San Francisco Estuary, California, USA. Estuar Coasts 35:603–621CrossRefGoogle Scholar
  7. Cachot J, Geffard O, Augagneur S, Lacroix S, Le Menach K, Peluhet L, Couteau J, Denier X, Devier MH, Pottier D, Budzinski H (2006) Evidence of genotoxicity related to high PAH content of sediments in the upper part of the Seine estuary (Normandy, France). Aquat Toxicol 79:257–267CrossRefGoogle Scholar
  8. Cachot J, Law M, Pottier D, Peluhet L, Norris M, Budzinski H, Winn R (2007) Characterization of toxic effects of sediment-associated organic pollutants using the λ transgenic medaka. Environ Sci Technol 41:7830–7836CrossRefGoogle Scholar
  9. Cailleaud K, Forget-Leray J, Souissi S, Hilde D, LeMenach K, Budzinski H (2007) Seasonal variations of hydrophobic organic contaminant concentrations in the water-column of the Seine Estuary and their transfer to a planktonic species Eurytemora affinis (Calanoïda, copepoda). Part 1: PCBs and PAHs. Chemosphere 70:270–280CrossRefGoogle Scholar
  10. Cailleaud K, Forget-Leray J, Peluhet L, LeMenach K, Souissi S, Budzinski H (2009) Tidal influence on the distribution of hydrophobic organic contaminants in the Seine estuary and biomarker responses on the copepod Eurytemora affinis. Environ Pollut 157:64–71CrossRefGoogle Scholar
  11. Cooke JB, Hinton DE (1999) Promotion by 17β-estradiol and β-hexachlorocyclohexane of hepatocellular tumors in medaka, Oryzias latipes. Aquat Toxicol 45:127–145CrossRefGoogle Scholar
  12. Dezfuli BS, Pironi F, Shinn AP, Manera M, Giari L (2007) Histopathology and ultrastructure of Platichthys flesus naturally infected with Anisakis simplex s.l. larvae (Nematoda: Anisakidae). J Parasitol 93:1416–1423CrossRefGoogle Scholar
  13. El-Darsh HEM, Whitfield PJ (1999) The parasite community infecting flounders, Platichthys flesus, in the tidal Thames. J Helminthol 73:203–214Google Scholar
  14. Ericson G, Lindesjöö E, Balk L (1998) DNA adducts and histopathological lesions in perch (Perca fluviatilis) and northern pike (Esox lucius) along a polycyclic aromatic hydrocarbon gradient on the Swedish coastline of the Baltic Sea. Can J Fish Aquat Sci 55:815–824CrossRefGoogle Scholar
  15. Feist SW, Lang T, Stentiford GD, Köhler A (2004) Biological effects of contaminants: use of liver pathology of the European flatfish dab (Limanda limanda L.) and flounder (Platichthys flesus L.) for monitoring. ICES Techniques in Marine Environmental Sciences 38, 42 ppGoogle Scholar
  16. Gilliers C, Le Pape O, Désaunay Y, Morin J, Guérault D, Amara R (2006) Are growth and density quantitative indicators of essential fish habitat quality? An application to the common sole Solea solea nursery grounds. Estuar Coast Self Sci 69:96–106CrossRefGoogle Scholar
  17. Harmon SM, Wiley FE (2011) Effects of pollution on freshwater organisms. Water Environ Res 83:1733–1788CrossRefGoogle Scholar
  18. Harris JRW, Cleary JJ, Valkirs AO (1996) Particle–water partitioning and the role of sediments as a sink and secondary source of TBT. In: Champ MA, Seligman PF (eds) Organtin. Environmental fate and effects. Chapman & Hall, London, pp 459–474Google Scholar
  19. Harshbarger JC, Clark JB (1990) Epizootiology of neoplasms in bony fish of North America. Sci Total Environ 94:1–32CrossRefGoogle Scholar
  20. Hinrichsen D (1998) In: Hinrichsen D (ed) Coastal waters of the world. Trends, threats and strategies. Island Press, WashingtonGoogle Scholar
  21. Hinton DE, Segner H, Braunbeck T (2001) Toxic responses of the liver. In: Schlenk D, Bensen WH (eds) Toxicity in marine and freshwater teleosts, vol. 1. Taylor and Francis, London, pp 224–268Google Scholar
  22. Johnson LL, Stehr CM, Olson OP, Myers MS, Pierce SP, Wigren CA, McCain BB, Varanasi U (1993) Chemical contaminants and hepatic lesions in winter flounder (Pleuronectes americanus) from the Northeast Coast of the United States. Environ Sci Technol 27:2759–2771CrossRefGoogle Scholar
  23. Koehler A (2004) The gender-specific risk to liver toxicity and cancer of flounder (Platichthys flesus L.) at the German Wadden Sea coast. Aquat Toxicol 70:257–276CrossRefGoogle Scholar
  24. Lang T, Wosniok W, Baršiene J, Broeg K, Kopecka J, Parkkonen J (2006) Liver histopathology in Baltic flounder (Platichthys flesus) as indicator of biological effects of contaminants. Mar Pollut Bull 53:488–496CrossRefGoogle Scholar
  25. Laroche J, Gauthier O, Quiniou L, Devaux A, Bony S, Evrard E, Cachot J, Cherel Y, Larcher T, Riso R, Pichereau V, Devier M-H, Budzinski H (2012) Variation patterns in individual fish responses to chemical stress among estuaries, seasons and genders: the case of the European flounder (Platichthys flesus) in the Bay of Biscay. Environ Sci Pol Res (this issue)Google Scholar
  26. Lehman PW, Boyer G, Satchwell M, Waller S (2008) The influence of environmental conditions on the seasonal variation of microcystis cell density and microcystins concentration in San Francisco Estuary. Hydrobiologia 600:187–204CrossRefGoogle Scholar
  27. Lyons BP, Stewart C, Kirby MF (1999) The detection of biomarkers of genotoxin exposure in the European flounder (Platichthys flesus) collected from the River Tyne Estuary. Mutat Res Genet Toxicol Environ Mutagen 446:111–119CrossRefGoogle Scholar
  28. Lyons BP, Stentiford GD, Green M, Bignell J, Bateman K, Feist SW, Goodsir F, Reynolds WJ, Thain JE (2004) DNA adduct analysis and histopathological biomarkers in European flounder (Platichthys flesus) sampled from UK estuaries. Mutat Res Fundam Mol Mech Mutagen 552:177–186CrossRefGoogle Scholar
  29. Malmström CM, Miettinen S, Bylund G (2000) DNA adducts in liver and leukocytes of flounder (Platichthys flesus) experimentally exposed to benzo[a]pyrene. Aquat Toxicol 48:177–184CrossRefGoogle Scholar
  30. Marchand J, Tanguy A, Laroche J, Quiniou L, Moraga D (2003) Responses of European flounder Platichthys flesus populations to contamination in different estuaries along the Atlantic coast of France. Mar Ecol Prog Ser 260:273–284CrossRefGoogle Scholar
  31. Marchand J, Quiniou L, Riso R, Thebaut M-T, Laroche J (2004) Physiological cost of tolerance to toxicants in the European flounder Platichthys flesus, along the French Atlantic Coast. Aquat Toxicol 70:327–343CrossRefGoogle Scholar
  32. Mearns AJ, Reish DJ, Oshida PS, Ginn T, Rempel-Hester MA (2011) Effects of pollution on marine organisms. Water Environ Res 83:1789–1852CrossRefGoogle Scholar
  33. Miller EC, Miller JA (1981) Mechanisms of chemical carcinogenesis. Cancer Res 47:1055–1064Google Scholar
  34. Minier C, Abarnou A, Jaouen-Madoulet A, Le Guellec A-M, Tutundjian R, Bocquené G, Leboulenger F (2006) A pollution-monitoring pilot study involving contaminant and biomarker measurements in the Seine Estuary, France, using zebra mussels (Dreissena polymorpha). Environ Toxicol Chem 25:112–119CrossRefGoogle Scholar
  35. Myers MS, Rhodes LD, McCain BB (1987) Pathologic anatomy and patterns of occurrence of hepatic neoplasms, putative preneoplastic lesions, and other idiopathic hepatic conditions in English sole (Parophrys vetulus) from Puget Sound, Washington. J Natl Cancer Inst 78:333–363Google Scholar
  36. Myers MS, Johnson LL, Olson OP, Stehr CM, Horness BH, Collier TK, Mccain BB (1998) Toxicopathic hepatic lesions as biomarkers of chemical contaminant exposure and effects in marine bottom fish species from the Northeast and Pacific Coasts, USA. Mar Pollut Bull 37:92–113CrossRefGoogle Scholar
  37. Myers MS, Johnson LL, Collier TK (2003) Establishing the causal relationship between polycyclic aromatic hydrocarbon (PAH) exposure and hepatic neoplasms and neoplasia-related liver lesions in English sole (Pleuronectes vetulus). Hum Ecol Risk Assess 9:67–94CrossRefGoogle Scholar
  38. Nunez O, Hendricks JD, Arbogast DN, Fong AT, Lee BC, Bailey GS (1989) Promotion of aflatoxin B1 hepatocarcinogenesis in rainbow trout by 17-β-estradiol. Aquat Toxicol 15:289–302CrossRefGoogle Scholar
  39. Reddy MV, Randerath K (1986) Nuclease P1-mediated enhancement of sensitivity of 32P-postlabeling test for structurally diverse DNA adducts. Carcinogenesis 7:1543–1551CrossRefGoogle Scholar
  40. Rocher B, Le Goff J, Briand M, Manduzio H, Peluhet L, Gallois J, Devier M-H, Gricourt L, Augagneur S, Budzinski H, Pottier D, Andrée V, Lebailly P, Cachot J (2006) Genotoxicant accumulation and cellular defence activation in bivalves chronically exposed to waterborne contaminants from the Seine River. Aquat Toxicol 79:65–77CrossRefGoogle Scholar
  41. Rose WL, French BL, Reichert WL, Faisal M (2000) DNA adducts in hematopoietic tissues and blood of the mummichog (Fundulus heteroclitus) from a creosote-contaminated site in the Elizabeth River, Virginia. Mar Environ Res 50:581–589CrossRefGoogle Scholar
  42. SGIMC (2011) Report of the Joint ICES/OSPAR Study Group on Integrated Monitoring of Contaminants and Biological Effects (SGIMC), 14–18 March 2011, Copenhagen, Denmark, ICES Advisory Committee: ICES CM 2011/ACOM:30Google Scholar
  43. Shaw GR, Connell DW (2001) DNA adducts as a biomarker of polycyclic aromatic hydrocarbon exposure in aquatic organisms: relationship to carcinogenicity. Biomarkers 6:64–71CrossRefGoogle Scholar
  44. Stentiford GD, Longshaw M, Lyons BP, Jones G, Green M, Feist SW (2003) Histopathological biomarkers in estuarine fish species for the assessment of biological effects of contaminants. Mar Environ Res 55:137–159CrossRefGoogle Scholar
  45. Varanasi U, Reichert WL, Stein JE (1989) 32P-postlabeling analysis of DNA adducts in liver of wild English sole (Parophrys vetulus) and winter flounder (Pseudopleuronectes americanus). Cancer Res 49:1171–1177Google Scholar
  46. Vethaak AD (1992) Diseases of flounder (Platichthys flesus L.) in the Dutch Wadden Sea, and their relation to stress factors. Neth J Sea Res 29:257–272CrossRefGoogle Scholar
  47. Vethaak AD, Jol JG, Meijboom A, Eggens ML, Rheinallt TA, Wester PW, Van De Zande T, Bergman A, Dankers N, Ariese F, Baan RA, Everts JM, Opperhuizen A, Marquenie JM (1996) Skin and liver diseases induced in flounder (Platichthys flesus) after long-term exposure to contaminated sediments in large-scale mesocosms. Environ Health Perspect 104:1218–1229CrossRefGoogle Scholar
  48. Vethaak AD, Jol JG, Martinez-Gomez C (2011) Effects of cumulative stress on fish health near freshwater outlet sluices into the sea: a case study (1988–2005) with evidence for a contributing role of chemical contaminants. Integr Environ Assess Manag 7:445–458CrossRefGoogle Scholar
  49. Vetkaak AD, Wester PW (1996) Diseases of flounder Platichthys flesus in Dutch coastal and estuarine waters, with particular reference to environmental stress factors. II. Liver histopathology. Dis Aqua Org 26:99–116CrossRefGoogle Scholar
  50. Williams TD, Turan N, Diab AM, Wu H, Mackenzie C, Bartie KL, Hrydziuszko O, Lyons BP, Stentiford Grant D, Herbert JM, Abraham JK, Katsiadaki I, Leaver MJ, Taggart JB, George SG, Viant MR, Chipman KJ, Falcian F (2011) Towards a system level understanding of non-model organisms sampled from the environment: a network biology approach. PLoS Comput Biol 7:1–20CrossRefGoogle Scholar
  51. Wolf J, Wolfe MJ (2005) A brief overview of nonneoplastic hepatic toxicity in fish. Toxicol Pathol 33:75–85CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Jérôme Cachot
    • 1
    Email author
  • Yan Cherel
    • 2
  • Thibaut Larcher
    • 2
  • Annie Pfohl-Leszkowicz
    • 3
  • Jean Laroche
    • 4
  • Louis Quiniou
    • 4
  • Jocelyne Morin
    • 5
  • Julien Schmitz
    • 6
  • Thierry Burgeot
    • 7
  • Didier Pottier
    • 8
  1. 1.Univ. Bordeaux, EPOC UMR 5805, LPTC GroupTalenceFrance
  2. 2.INRA, UMR 703, Oniris, La ChantrerieNantesFrance
  3. 3.Univ. Toulouse, Laboratoire de Génie Chimique, UMR CNRS/INPT/UPS 5503, INP/ENSA ToulouseCastanet-TolosanFrance
  4. 4.Université de Brest, UMR 6539, LEMAR, Institut Universitaire Européen de la MerPlouzanéFrance
  5. 5.Ifremer, Laboratoire Ressources HalieutiquesPort-en-BessinFrance
  6. 6.Institut Français du PétroleRueil-Malmaison CedexFrance
  7. 7.Ifremer Nantes, Département Polluants ChimiquesNantes CedexFrance
  8. 8.Université de Caen Basse-Normandie, UR ABTE EA 4651, Centre François BaclesseCaenFrance

Personalised recommendations