Target organs of the Manila clam Ruditapes philippinarum for studying metal accumulation and biomarkers in pollution monitoring: laboratory and in-situ transplantation experiments

  • Eun-Ji Won
  • Kyung-Tae Kim
  • Jin-Young Choi
  • Eun-Soo Kim
  • Kongtae RaEmail author


To characterize the target organs of the Manila clam Ruditapes philippinarum for use in environmental study, the accumulation of trace metals and three biomarkers was measured in different organs. Exposure with Cu and Pb carried out under laboratory conditions revealed a linear uptake of metals throughout the experimental period in each tissue. In particular, significant increase was observed in gills and mantle. The increase of intracellular reactive oxygen species showed the great potential of gills as a target tissue for both Cu and Pb exposure. The highest activity of glutathione S-transferase and their relative increase in activity were also observed in gills. Metallothionein-like protein levels, however, increased greatly in the digestive gland and mantle during Cu and Pb exposure, respectively, although all tissues, except the foot, showed significant changes after 24 h of metal exposure. In the field study, the highest concentration of metals was recorded in the gills and mantle, accounting for over 50 % of the total accumulated metal in all sites. Additionally, Cu and Pb increased significantly in these two organs, respectively. However, the order of accumulation rate in laboratory exposure was not concomitant with those of the lab-based study, suggesting that different routes of metal uptake and exposure duration induce distinct partitioning of metals and regulating system in R. philippinarum. These series of exposure studies demonstrated that gills, mantle, and digestive gland in R. philippinarum are potential target tissues in environmental monitoring study using metal concentrations and biomarkers.


Manila clam Ruditapes philippinarum Trace metal Bioaccumulation Reactive oxygen species Glutathione S-transferase Metallothionein-like proteins (MTLPs) 



This research was supported by a grant from the KIOST [PE99402] funded to Kongtae Ra and partially supported by grant from the National Research Foundation of Korea (NRF-2015R1C1A2A01053437) funded to Eun-Ji Won.

Supplementary material

10661_2016_5485_MOESM1_ESM.docx (10.4 mb)
ESM 1 (DOCX 10622 kb)


  1. Ahn, I.-Y., Ji, J., Choi, H. J., Pyu, S.-H., Park, H., Choi, J.-W., et al. (2006). Spatial variation of heavy metal accumulation in Manila clam Ruditapes philippinarum from some selected intertidal flats of Korea. Ocean and Polar Research, 28, 215–224.CrossRefGoogle Scholar
  2. Alibabić, V., Vahčić, N., Bajramović, M., et al. (2007). Bioaccumulation of metals in fish of salmonidae family and the impact on fish meat quality. Environmental Monitoring and Assessment, 131(1), 349–364.CrossRefGoogle Scholar
  3. Boalt, E., Dahlgren, H., Miller, A., et al. (2011). Cadmium, lead and mercury concentrations in whole fish, liver and muscle of herring (Clupea harengus) and perch (Perca fluviatilis). Report written for Naturvårdsverket (Swedish Environmental Protection Agency). Report Nr 6:2012. 10 pp.Google Scholar
  4. Boening, D. W. (1999). An evaluation of bivalves as biomonitors of heavy metals pollution in marine waters. Environmental Monitoring and Assessment, 55, 495–470.CrossRefGoogle Scholar
  5. Bonneris, E., Perceval, O., Masson, S., Hare, L., Campbell, P. G., et al. (2005). Sub-cellular partitioning of Cd, Cu and Zn in tissues of indigenous unionid bivalves living along a metal exposure gradient and links to metal–induced effects. Environmental Pollution, 135, 195–208.CrossRefGoogle Scholar
  6. Burger, J., Gochfeld, M., Jeitner, C., Burke, S., Stamm, T., et al. (2007). Metal levels in flathead sole (Hippoglossoides elassodon) and great sculpin (Myoxocephalus polyacanthocephalus) from Adak Island, Alaska: potential risk to predators and fishermen. Environmental Research, 103(1), 62–69.CrossRefGoogle Scholar
  7. Chen, C. Y., Stemberger, R. S., Klaue, B., Blum, J. D., Pickhardt, P. C., Folt, C. L., et al. (2000). Accumulation of heavy metals in food web components across a gradient of lakes. Limnology and Oceanography, 45(7), 1525–1536.CrossRefGoogle Scholar
  8. Choi, J. Y., Yu, J., Yang, D. B., Ra, K., Kim, K. T., Hong, G. H., Shin, K. H., et al. (2011). Acetylthiocholine (ATC)-cleaving cholinesterase (ChE) activity as a potential biomarker of pesticide exposure in the Manila clam, Ruditapes philippinarum, of Korea. Marine Environmental Research, 71, 1162–1168.CrossRefGoogle Scholar
  9. Copper, S., Hare, L., Campbell, P. G., et al. (2010). Subcellular partitioning of cadmium in the freshwater bivalve, Pyganodon grandis, after separate short-term exposures to waterborne or diet-borne metal. Aquatic Toxicology, 100, 303–312.CrossRefGoogle Scholar
  10. Dallinger, R., Berger, B., Hunziker, P., Kägi, J. H. R., et al. (1997). Metallothionein in snail Cd and Cu metabolism. Nature, 388, 237–238.CrossRefGoogle Scholar
  11. De Luca-Abbott, S. B., Richardson, B. J., McCellan, K. E., Zheng, G. J., Martin, M., Lam, P. K. S., et al. (2005). Field validation of antioxidant enzyme biomarkers in mussels (Perna viridis) and clams (Ruditapes philippinarum) transplanted in Hong Kong coastal water. Marine Pollution Bulletin, 51, 694–707.CrossRefGoogle Scholar
  12. Depledge, M. H., Aagaard, A., Györkös, P., et al. (1995). Assessment of trace metal toxicity using molecular, physiological and behavioral biomarkers. Marine Pollution Bulletin, 21, 19–27.CrossRefGoogle Scholar
  13. Fernández-Tajes, J., Flórez, F., Pereira, S., Rábade, T., Laffon, B., Méndez, J., et al. (2011). Use of three bivalve species for biomonitoring a polluted estuarine environment. Environmental Monitoring and Assessment, 177, 1–4.CrossRefGoogle Scholar
  14. Gabr, H. R., Gab-Alla, A. A.-F. A., et al. (2008). Effect of transplantation on heavy metal concentrations in commercial clams of Lake Timsah, Suez Canal, Egypt. Oceanologia, 50(1), 83–93.Google Scholar
  15. Jezierska, B., & Witeska, M. (2006). The metal uptake and accumulation in fish living in polluted waters. Soil and Water Pollution Monitoring, Protection and Remediation, 69, 3–23.CrossRefGoogle Scholar
  16. Ji, J., Choi, H. J., Ahn, I.-Y., et al. (2006). Evaluation of Manila clam Ruditapes philippinarum as a sentinel species for metal pollution monitoring in estuarine tidal flats of Korea: effects of size, sex, and spawning on baseline accumulation. Marine Pollution Bulletin, 52, 447–453.CrossRefGoogle Scholar
  17. Kim, K.-T., Kim, E. S., Cho, S. R., Park, J. K., Park, C. K., et al. (2003). Change of heavy metals in the surface sediments of the Lake Shihwa and its tributaries. Ocean Polar Research, 25, 447–457.CrossRefGoogle Scholar
  18. Luxama, J. D., Carroll, M. A., Catapane, E. J., et al. (2010). Effects of potential therapeutic agents on copper accumulations in gill of Crassostrea virginica. In Vivo, 31, 32–42.Google Scholar
  19. Martel, P., Kovacs, T., Voss, R., Megraw, S., et al. (2003). Evaluation of caged freshwater mussels as an alternative method for environmental effects monitoring (EEM) studies. Environmental Pollution, 124, 471–483.CrossRefGoogle Scholar
  20. Martín-Díaz, M. L., Blasco, J., Sales, D., DelValls, T. A., et al. (2007). Biomarkers study for sediment quality assessment in Spanish ports using the crab Carcinus maenas and the clam Ruditapes philippinarum. Archives of Environmental Contamination and Toxicology, 53, 66–76.CrossRefGoogle Scholar
  21. Maruya, K. A., Dodder, N. G., Schaffner, R. A., Weisberg, S. B., Gregorio, D., Klosterhaus, S., Alvarez, D. A., Furlong, E. T., Kimbrough, K. L., Lauenstein, G. G., Christensen, J. D., et al. (2013). Refocusing mussel watch on contaminants of emerging concern (CECs): the California pilot study (2009-10). Marine Pollution Bulletin, 81(2), 334–339.CrossRefGoogle Scholar
  22. Ra, K., Kim, K. T., Bang, J. H., Lee, J. M., Kim, E. S., Cho, S. R., et al. (2011). A preliminary study of environmental impact assessment of tidal power plant in Shihwa Lake, Korea: heavy metal accumulation in the transplanted Manila clam (Ruditapes philippinarum). Journal of Coastal Research, SI64, 932–936.Google Scholar
  23. Rainbow, P. S., & Dallinger, R. (1993). Metal uptake, regulation and excretion in freshwater invertebrates. In R. Dallinger & P. S. Rainbow (Eds.), Ecotoxicology of metals in invertebrates, SETAC special publication series (pp. 119–131). Boca Raton, FL: Lewis Publishers.Google Scholar
  24. Rainbow, P. S. (1995). Biomonitoring of heavy metal availability in the marine environment. Marine Pollution Bulletin, 31, 183–192.CrossRefGoogle Scholar
  25. Rainbbow, P. S. (2006). Biomonitoring of trace metals in estuarine and marine environment. Australasian Journal of Ecotoxicology, 12, 107–122.Google Scholar
  26. Regoli, F., Nigro, M., Bertoli, E., Principato, G., Orlando, E., et al. (1997). Defenses against oxidative stress in the Antarctic scallop Adamussium colbecki and effects of acute exposure to metals. Hydrobiologia, 355, 139–144.CrossRefGoogle Scholar
  27. Reynoldson, T. B. (1987). Interactions between sediment contaminants and benthic organisms. Hydrobiologia, 149, 53–66.CrossRefGoogle Scholar
  28. Romero-Ruiz, A., Amezcua, O., Rodríguez-Ortega, M. J., Muñoz, J. L., Alhama, J., Rodríguez-Ariza, A., Gómez-Ariza, J. L., López-Barea, J., et al. (2003). Oxidative stress biomarkers in bivalves transplanted to the Guadalquivir estuary after Aznalcóllar spill. Environmental Toxicology and Chemistry, 22, 92–100.CrossRefGoogle Scholar
  29. Saha, M., Sarkar, S. K., Bhattacharya, B., et al. (2006). Interspecific variation in heavy metal body concentrations in biota of Sunderban mangrove wetland, Northeast India. Environmental International, 32, 203–207.CrossRefGoogle Scholar
  30. Sarkar, S. K., Cabral, H., Cardoso, I., Bhattacharya, A. K., Satpathy, K. K., Alam, M. A., et al. (2008). Biomonitoring of heavy metals using the bivalve molluscs in Sunderban mangrove wetland, northeast coast of bay of Bengal (India): possible risks to human health. Clean, 36, 187–194.Google Scholar
  31. Szebedinszky, C., McGeer, J. C., McDonald, D. G., Wood, C. M., et al. (2001). Effects of chronic Cd exposure via the diet or water on internal organ-specific distribution and subsequent gill Cd uptake kinetics in juvenile rainbow trout (Oncorhynchus mykiss). Environmental Toxicology and Chemistry, 20, 597–607.CrossRefGoogle Scholar
  32. Tessier, A., & Campbell, P. G. C. (1987). Partitioning of trace metals in sediments: relationships with bioavailability. Hydrobiologia, 149, 43–52.CrossRefGoogle Scholar
  33. Valavanidis, A., Vlahogianni, T., Dassenakis, M., Scoullos, M., et al. (2006). Molecular biomarkers of oxidative stress in aquatic organisms in relation to toxic environmental pollutants. Ecotoxicology and Environmental Safety, 64, 178–189.CrossRefGoogle Scholar
  34. Viarengo, A., Ponzano, E., Dondero, F., Fabbra, R., et al. (1997). A simple spectrophotometric method for metallothionein evaluation in marine organisms: an application to Mediterranean and Antartic mollusk. Marine Environmental Research, 44, 69–84.CrossRefGoogle Scholar
  35. Wang, D., Couillard, Y., Campbell, G. C., Jolicoeur, P., et al. (2011). Changes in subcellular metal partitioning in the gills of freshwater bivalves (Pyganodon grandis) living along an environmental cadmium gradient. Canadian Journal of Fisheries and Aquatic Sciences, 56, 774–784.CrossRefGoogle Scholar
  36. Won, E.-J., Hong, S., Ra, K., Kim, K.-T., Shin, K.-H., et al. (2012). Evaluation of the potential impact of polluted sediments using Manila clam Ruditapes philippinarum: bioaccumulation and biomarker responses. Environmental Science and Pollution Research, 19, 2570–2580.CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2016

Authors and Affiliations

  • Eun-Ji Won
    • 1
  • Kyung-Tae Kim
    • 1
    • 2
  • Jin-Young Choi
    • 1
  • Eun-Soo Kim
    • 3
  • Kongtae Ra
    • 1
    • 2
    Email author
  1. 1.Marine Chemistry & Geochemistry Research CenterKorea Institute of Ocean Science and Technology (KIOST)AnsanRepublic of Korea
  2. 2.Department of Integrated Ocean SciencesKorea University of Science and Technology (UST)AnsanRepublic of Korea
  3. 3.Ocean Observation & Information SectionKorea Institute of Ocean Science and Technology (KIOST)AnsanRepublic of Korea

Personalised recommendations