Evaluation of Multixenobiotic Resistance in Dreissenid Mussels as a Screening Tool for Toxicity in Freshwater Sediments

  • A. Ács
  • K. Imre
  • Gy. Kiss
  • J. Csaba
  • J. Győri
  • Á. Vehovszky
  • A. FarkasEmail author


The multixenobiotic defense mechanism (MXR) in aquatic organisms was recognized as a first-line defense system, and its potential use as an early biomarker of exposure to environmental stress has raised attention in the last two decades. To evaluate the relevance of this biomarker in the freshwater mussel Dreissena polymorpha, we studied its responsiveness within laboratory exposures to contaminants sequestered in freshwater sediments affected by moderate anthropogenic impact. The effectiveness of this biomarker was assessed by comparing the MXR-transporter activities determined in bivalves first with toxicity scores recorded with the D. rerio embryo developmental assay. Both bioassays were applied in the sediment contact test format. As a second evaluation approach, MXR activities determined in exposed mussels were compared with sediment-contamination data integrated into toxic units on the basis of acute toxicity to Daphnia magna. In D. polymorpha subjected to acute exposure with moderately polluted sediments, we detected limited (22–33 %) but statistically significant induction of MXR activity. Mean MXR activities significantly correlated with TU values computed for test sediments. MXR activities in mussels showed strong positive correlation with the metal load of sediments and proved to be unrelated to the contamination with polycyclic aromatic compounds. MXR activity in laboratory-exposed mussels showed low variability within treatments and thus reliably reflected even low contaminant differences between the negative reference and moderately polluted harbor sediments. The strong correlation found in this study between the MXR-transporter activity in exposed mussels and environmentally realistic sediment contamination underscores the fairly good sensitivity of this biomarker in laboratory testing conditions to signal the bioavailability of sediment bound contaminants, and it may also anticipate even the incidence of toxicity to biota.


PAHs Hatching Success Freshwater Sediment Harbor Sediment Test Sediment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This research was supported by the European Union and the State of Hungary and cofinanced by the European Social Fund in the framework of TÁMOP-4.2.4.A/2-11/1-2012-0001 National Excellence Program. This work was supported by a grant from the Balaton Project of the Office of the Prime Minister of Hungary.


  1. Achard M, Baudrimont M, Boudou A, Bourdineaud JP (2004) Induction of a multixenobiotic resistance protein (MXR) in the Asiatic clam Corbicula fluminea after heavy metals exposure. Aquat Toxicol 67:347–357CrossRefGoogle Scholar
  2. Algaltoxkit FTM (1996) Freshwater toxicity test with microalgae. Standard operational procedure. Creasel, Deinze, p 28Google Scholar
  3. Austen MC, Somerfield PJ (1997) A community level sediment bioassay applied to an estuarine heavy metal gradient. Mar Environ Res 43:315–328CrossRefGoogle Scholar
  4. Bard SM (2000) Multixenobiotic resistance as a cellular defense mechanism in aquatic organisms. Aquat Toxicol 48:357–389CrossRefGoogle Scholar
  5. Bard SM, Woodin BR, Stegeman JJ (2002) Expression of P-glycoprotein and cytochrome P450 1A in intertidal fish (Anoplarchus purpurescens) exposed to environmental contaminants. Aquat Toxicol 60(1–2):17–32CrossRefGoogle Scholar
  6. Behrens A, Segner H (2005) Cytochrome P4501A induction in brown trout exposed to small streams of an urbanized area: results of a five year study. Environ Pollut 136:231–242CrossRefGoogle Scholar
  7. Binelli A, Cogni D, Parolini M, Provini A (2010) Multi-biomarker approach to investigate the state of contamination of the R. Lambro/R. Po confluence (Italy) by zebra mussel (Dreissena polymorpha). Chemosphere 79:518–528CrossRefGoogle Scholar
  8. Bodin N, Burgeot T, Stanisiére JY, Bocquené G, Menard D, Minier C et al (2004) Seasonal variations of battery of biomarkers and physiological indices for the mussel Mytilus galloprovincialis transplanted into the northwest Mediterranean Sea. Comp Biochem Physiol 138:411–427Google Scholar
  9. Bodnár E, Polyák K, Hlavay J (2005) Material transport between the atmosphere and sediment of the Lake Balaton. Microchem J 79:221–230CrossRefGoogle Scholar
  10. Cajaraville MP, Bebianno MJ, Blasco J, Porte C, Sarasquete C, Viarengo A (2000) The use of biomarkers to assess the impact of pollution in coastal environments of the Iberian Peninsula: a practical approach. Sci Total Environ 247:295–311CrossRefGoogle Scholar
  11. Chapman PM, Ho KT, Munns WR, Soloman K, Weinstein MP (2002) Issues in sediment toxicity and ecological risk assessment. Mar Pollut Bull 44:271–278CrossRefGoogle Scholar
  12. Cleveland L, Little EE, Petty JD, Johnson BT, Lebo JA, Orazio CE et al (1997) Toxicological and chemical screening of Antarctica sediments: use of whole sediment toxicity tests, microtox, mutatox and semipermeable membrane devices (SPMDs). Mar Pollut Bull 34:194–202CrossRefGoogle Scholar
  13. Contardo-Jara V, Wiegand C (2008) Molecular biomarkers of Dreissena polymorpha for evaluation of renaturation success of a formerly sewage polluted stream. Environ Pollut 155:182–189CrossRefGoogle Scholar
  14. Contardo-Jara V, Lorenz C, Pflugmacher S, Nützmann G, Kloas W, Wiegeand C (2011) Exposure to human pharmaceuticals carbamazepine, ibuprofen and bezafibrate causes molecular effects in Dreissena polymorpha. Aquat Toxicol 105:208–437CrossRefGoogle Scholar
  15. Davoren M, Ni Shuilleabhain S, Halloran JO, Hartl MGJ, Sheehan D, O’Brien NM et al (2005) A test battery approach for the ecotoxicological evaluation of estuarine sediments. Ecotoxicology 14:741–755CrossRefGoogle Scholar
  16. Di Toro DM, Zarba CS, Hansen DJ, Berry WJ, Swartz RC et al (1991) Technical basis for establishing sediment quality criteria for nonionic organic chemicals by using equilibrium partitioning. Environ Toxicol Chem 10:1541–1583CrossRefGoogle Scholar
  17. Eertman RHM, Groenink CLFMG, Sandee B, Hummel H (1995) Response of the blue mussel Mytilus edutis L. following exposure to PAHs or contaminated sediments. Mar Environ Res 39:160–173CrossRefGoogle Scholar
  18. Epel D (1998) Use of multidrug transporters as a first line of defense against toxins in aquatic organisms. Comp Biochem Physiol A 120:23–28CrossRefGoogle Scholar
  19. Epel D, Luckenbach T, Stevenson CN, Macmanus-Spencer LA, Hamdoun A, Smital T (2008) Efflux transporters: newly appreciated roles in protection against pollutants. Environ Sci Tech 42:3914–3920CrossRefGoogle Scholar
  20. Eufemia NA, Epel D (2000) Introduction of the multixenobiotic defense mechanism (MXR), P-glycoprotein in the mussel Mytilus californianus as a general cellular response to environmental stress. Aquat Toxicol 49:89–100CrossRefGoogle Scholar
  21. Faria M, Navarro A, Luckenbach T, Piña B, Barata C (2011) Characterization of the multixenobiotic resistance (MXR) mechanism in embryos and larvae of the zebra mussel (Dreissena polymorpha) and studies on its role in tolerance to single and mixture combinations of toxicants. Aquat Toxicol 101:78–87CrossRefGoogle Scholar
  22. Franco JL, Trivella DBB, Trevisan R, Dinslaken DF, Marques MRF, Bainy ACD et al (2006) Antioxidant status and stress proteins in the gills of brown mussel Perna perna exposed to zinc. Chem Biol Interact 160:232–240CrossRefGoogle Scholar
  23. Garner LVT, Di Giulio RT (2012) Glutathione transferase pi class 2 (GSTp2) protects against the cardiac deformities caused by exposure to PAHs but not PCB-126 in zebrafish embryos. Comp Biochem Physiol C 155(4):573–579Google Scholar
  24. Ghirardini AV, Birkenmeyer T, Novelli AA, Delaney E, Pavoni B, Ghetti PF (1999) An integrated approach to sediment quality assessment: the Venetian lagoon as a case study. Aquat Ecosyst Health 2:435–437CrossRefGoogle Scholar
  25. Guerlet E, Ledy K, Meyer A, Giamberini L (2007) Towards a validation of a cellular biomarker suite in native and transplanted zebra mussels: a 2-year integrative field study of seasonal and pollution-induced variations. Aquat Toxicol 81:377–388CrossRefGoogle Scholar
  26. Gy Kiss, Gelencsér A, Krivácsy Z, Hlavay J (1997) Occurrence and determination of organic pollutants in aerosol, precipitation, and sediment samples collected at Lake Balaton. J Chromatogr A 774(1–2):349–361Google Scholar
  27. Hlavay J, Polyák K (2002) Investigation on the pollution sources of bottom sediments in the Lake Balaton. Microchem J 73(1–2):65–78CrossRefGoogle Scholar
  28. Höss S, Claus E, Von der Ohe PC, Brinke M, Güde H, Heininger P et al (2011) Nematode species at risk—a metric to assess pollution in soft sediments of freshwaters. Environ Int 37(5):940–949CrossRefGoogle Scholar
  29. Incardona JP, Collier TK, Scholz NL (2004) Defects in cardiac function precede morphological abnormalities in fish embryos exposed to polycyclic aromatic hydrocarbons. Toxicol Appl Pharmacol 196(2):191–205CrossRefGoogle Scholar
  30. Ingersoll CG, Brunson EL, Dwyer FJ, Ankley JT, Benoit DA, Norberg-King TJ et al (1995) Toxicity and bioaccumulation of sediment-associated contaminants using freshwater invertebrates: a review of methods and applications. Environ Toxicol Chem 14:1885–1894CrossRefGoogle Scholar
  31. Ingersoll CG, Dillion T, Bildinger GR (1997) Ecological risk assessment of contaminated sediments. SETAC Press, PensacolaGoogle Scholar
  32. Kurelec B (1992) The multixenobiotic resistance mechanism in aquatic organisms. Crit Rev Toxicol 22:23–43CrossRefGoogle Scholar
  33. Lappalainen J, Juvonen R, Vaajasaari K, Karp M (1999) A new flash method for measuring the toxicity of solid and colored samples. Chemosphere 38(5):1069–1083CrossRefGoogle Scholar
  34. Lekube X, Izagirre U, Soto M, Marigómez I (2014) Lysosomal and tissue-level biomarkers in mussels cross-transplanted among four estuaries with different pollution levels. Sci Total Environ 472:36–48CrossRefGoogle Scholar
  35. Luckenbach T, Epel D (2008) ABCB- and ABCC-type transporters confer multixenobiotic resistance and form an environment-tissue barrier in bivalve gills. Am J Physiol Regul Integr Comp Physiol 294:R1919–R1929CrossRefGoogle Scholar
  36. Luckenbach T, Triesbskorn R, Müller E, Oberemm A (2001) Toxicity of waters from two streams to early life stages of brown trout (Salmo trutta f. fario L.) tested under semi-field conditions. Chemosphere 45:571–579CrossRefGoogle Scholar
  37. Lüdeking A, Köhler A (2002) Identification of six mRNA sequences of genes related to multixenobiotic resistance (MXR) and biotransformation in Mytilus edulis. Mar Ecol Prog Ser 238:115–124CrossRefGoogle Scholar
  38. Lüdeking A, Köhler A (2004) Regulation of expression of multixenobiotic resistance (MXR) genes by environmental factors in the blue mussel Mytilus edulis. Aquat Toxicol 69:1–10CrossRefGoogle Scholar
  39. McDonald DD, Ingersoll CG, Berger TA (2000) Development and evaluation of consensus-based sediment quality guidelines for freshwater ecosystems. Arch Environ Contam Toxicol 39:20–31CrossRefGoogle Scholar
  40. Minier C, Borghi V, Moore MN, Porte C (2000) Seasonal variation of MRX and stress proteins in the common mussel Mytilus galloprovincialis. Aquat Toxicol 50(3):167–176CrossRefGoogle Scholar
  41. Navarro A, Weißbach S, Faria M, Barata C, Piña B, Luckenbach T (2012) Abcb and Abcc transporter homologs are expressed and active in larvae and adults of zebra mussel and induced by chemical stress. Aquat Toxicol 122–123:144–152CrossRefGoogle Scholar
  42. Nebeker AV, Miller CE (1988) Use of the Amphipod Crustacean Hyalella azteca in freshwater and estuarine sediment toxicity tests. Environ Toxicol Chem 7:1027–1033CrossRefGoogle Scholar
  43. Nguyen HL, Leermakers M, Osán J, Török S, Baeyens W (2005) Heavy metals in Lake Balaton: water column, suspended matter, sediment and biota. Sci Total Environ 340(1–3):213–230CrossRefGoogle Scholar
  44. Pain S, Parant M (2007) Identification of multixenobiotic defence mechanism (MXR) background activities in the freshwater bivalve Dreissena polymorpha as reference values for its use as biomarker in contaminated ecosystems. Chemosphere 67:1258–1263CrossRefGoogle Scholar
  45. Pampanin DM, Marangon I, Volpato E, Campesan G, Nasci C (2005) Stress biomarkers and alkali-labile phosphate level in mussels (Mytilus galloprovincialis) collected in the urban area of Venice (Venice Lagoon, Italy). Environ Pollut 136(1):103–107CrossRefGoogle Scholar
  46. Pederson F, Bjørnestad E, Andersen HV, Kjølholt J, Poll C (1998) Characterization of sediments from Copenhagen harbor by use of biotests. Water Sci Technol 37:233–240CrossRefGoogle Scholar
  47. Phipps GL, Ankley GT, Benoit DA, Mattson VR (1993) Use of the aquatic oligochaete lumbriculus variegatus for assessing the toxicity and bioaccumulation of sediment-associated contaminants. Environ Toxicol Chem 12:269–274CrossRefGoogle Scholar
  48. Quinn B, Gagné F, Costello M, McKenzie K, Wilson J, Mothersill C (2004) The endocrine disrupting effects of municipal effluent on the zebra mussel (Dreissena polymorpha). Aquat Toxicol 66:279–292CrossRefGoogle Scholar
  49. Smital T, Kurelec B (1998) The activity of multixenobiotic resistance mechanism determined by rhodamine B-efflux method as a biomarker of exposure. Mar Environ Res 46:443–447CrossRefGoogle Scholar
  50. Smital T, Sauerborn R, Pivcević B, Krca S, Kurelec B (2000) Interspecies differences in P-glycoprotein mediated activity of multixenobiotic resistance mechanism in several marine and freshwater invertebrates. Comp Biochem Physiol C 126:175–186Google Scholar
  51. Smital T, Sauerborn R, Hackenberger BK (2003) Inducibility of the P-glycoprotein transport activity in the marine mussel Mytilus galloprovincialis and the freshwater mussel Dreissena polymorpha. Aquat Toxicol 65:443–465CrossRefGoogle Scholar
  52. Sprague JB (1970) Measurement of pollutant toxicity to fish. II—Utilizing and applying bioassay 467 results. Water Res 4:3–32CrossRefGoogle Scholar
  53. Sundt RC, Pampanin DM, Grung M, Baršienė J, Ruus A (2011) PAH body burden and biomarker responses in mussels (Mytilus edulis) exposed to produced water from a North Sea oil field: laboratory and field assessments. Mar Pollut Bull 62:1498–1505CrossRefGoogle Scholar
  54. Tuikka AI, Schmitt C, Höss S, Bandow N, Von der Ohe PC, De Zwart D et al (2011) Toxicity assessment of sediments from three European river basins using a sediment contact test battery. Ecotoxicol Environ Saf 74:123–131CrossRefGoogle Scholar
  55. Van Der Kooij LA, Van De Meent D, Van Leeuwen CJ, Bruggeman WA (1991) Deriving quality criteria for water and sediment from the results of aquatic toxicity tests and product standards: application of the equilibrium partitioning method. Water Res 25:697–705CrossRefGoogle Scholar
  56. Von der Ohe PC, Liess M (2004) Relative sensitivity distribution of aquatic invertebrates to organic and metal compounds. Environ Toxicol Chem 23:150–156CrossRefGoogle Scholar
  57. Von der Ohe PC, Deckere E, Prüß A, Munoz I, Wolfram G, Villagrasa M et al (2009) Toward an integrated assessment of the ecological and chemical status of European river basins. Integr Environ Assess Manag 5:50–61CrossRefGoogle Scholar
  58. Wolfram G, Höss S, Ordent C, Schmitt C, Adámek Z, Bandow N et al (2012) Assessing the impact of chemical pollution on benthic invertebrates from three different European rivers using a weight-of-evidence approach. Sci Total Environ 438:498–509CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • A. Ács
    • 1
    • 2
  • K. Imre
    • 3
  • Gy. Kiss
    • 3
  • J. Csaba
    • 1
  • J. Győri
    • 1
  • Á. Vehovszky
    • 1
  • A. Farkas
    • 1
    Email author
  1. 1.MTA ÖK Centre for Ecological ResearchBalaton Limnological InstituteTihanyHungary
  2. 2.Department of LimnologyUniversity of PannoniaVeszprémHungary
  3. 3.MTA-PE Air Chemistry Research GroupVeszprémHungary

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