Advertisement

Ecotoxicology

, Volume 22, Issue 3, pp 433–445 | Cite as

In situ exposure history modulates the molecular responses to carbamate fungicide Tattoo in bivalve mollusk

  • Halina I. Falfushynska
  • Lesya L. Gnatyshyna
  • Oksana B. StoliarEmail author
Article

Abstract

The aim of the present study was the investigation of the effect of in situ exposure history on the responses of freshwater mussels to thiocarbamate fungicide. Male bivalve mollusks Anodonta anatina (Unionidae) from polluted (A) and unpolluted (F) sites were subjected to 14 days of exposure to fungicide Tattoo (mixture of propamocarb and mancozeb, 91 μg L−1). When unexposed mussels were compared, chronic effect of toxic environment in site A was confirmed by oxidative stress indices (high levels of superoxide dismutase and catalase activities, lipid peroxidation, protein carbonyls and oxyradical production, low level of total glutathione (GSH)), genotoxicity (high levels of DNA-strand breaks and caspase-3 activity in digestive gland), and cytotoxicity (low lysosomal membrane stability in hemocytes), elevated vitellogenin-like proteins (Vtg-LP) concentration in gonads, high levels of Cu, Zn, Cd, metallothionein (MT)-bound metals (MT-Me) and MT-related thiol (MT-SH), and low ethoxyresorufin-O-deethylase (EROD) activity in digestive gland. The major differences in the responses of the two exposed groups were related to antioxidant defense and MT: in the group A, prominent oxidative stress response with the participation of MT-SH and GSH in the gills, EROD activation, but decrease of MT-Me level was shown, whereas in group F exposure provoked the elevation of MT-Me, caspase-3 and Vtg-LP values. Carbamate did not cause cholinesterase depletion and cytotoxicity. However, genotoxic and pro-oxidant effects (increased levels of hemocytes with micronuclei and nuclear abnormalities, DNA-strand breaks and oxyradical in digestive gland), were common responses for both the exposed groups.

Keywords

Bivalve mollusk Thiocarbamate In situ exposure history Metallothionein Oxidative stress Toxicity 

Notes

Acknowledgments

This work was Granted by Ministry of Education and Science, Youth and Sports of Ukraine and State Fund of Fundamental Research (Ukrainian-Hungarian (M/25-2009, M/25-2011), the application-oriented project of Ministry of Education and Science, Youth and Sport of Ukraine # 118B; R&D Project GP/F32/025 # F32/202-2011, and partly supported by West-Ukrainian BioMedical Research Center. The authors are grateful Post-graduate student O. Turta, students O. Goryn, I. Gricay, M. Zhuk, H. Klantca, State Agency of environment protection and State Agency of Fishery Supervision in Ternopil Region for the technical assistance, to Global Edico and Dr B. Pechenyak for the linguistic and phraseological improvement of this manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Aebi H (1974) Catalase. In: Bergmeyer HU (ed) Meth enzym anal. Academic Press, London, pp 671–684Google Scholar
  2. Ait Alla A, Mouneyrac C, Durou C, Moukrim A, Pellerin J (2006) Tolerance and biomarkers as useful tools for assessing environmental quality in the Oued Souss estuary (Bay of Agadir, Morocco). Comp Biochem Physiol 143C:23–29Google Scholar
  3. Anderson ME (1985) Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol 113:548–555CrossRefGoogle Scholar
  4. Augustyniak M, Babczynska A, Migula P, Wilczek G, Laszczyca P, Kafel A, Augustyniak M (2005) Joint effects of dimethoate and heavy metals on metabolic responses in a grasshopper (Chorthippus brunneus) from a heavy metals pollution gradient. Comp Biochem Physiol 141(4):412–419Google Scholar
  5. Augustyniak M, Babczynska A, Augustyniak M (2009) Does the grasshopper Chorthippus brunneus adapt to metal polluted habitats? a study of glutathione-dependent enzymes in grasshopper nymphs. Insect Sci 16:33–42CrossRefGoogle Scholar
  6. Bacanskas LR, Whitaker J, Di Giulio RT (2004) Oxidative stress in two populations of killifish (Fundulus heteroclitus) with differing contaminant exposure histories. Mar Environ Res 58:597–601CrossRefGoogle Scholar
  7. Ballan-Dufrançais C, Jeantet AY, Geffard A, Amiard JC, Amiard-Triquet C (2001) Cellular and tissular distribution of copper in an intrasedimentary bivalve, the Baltic clam Macoma balthica, originating from a clean or a metal-rich site. Can J Fish Aquat Sci 58:1964–1974CrossRefGoogle Scholar
  8. Barsiene J, Andreikenaite L, Rybakovas A (2006) Cytogenetic damage in perch (Perca fluviatilis L.) and duck mussel (Anodonta anatina L.) exposed to crude oil. Ekologija 1:25–31Google Scholar
  9. Barsiene J, Andreikėnaitė L, Garnaga G, Rybakovas A (2008) Genotoxic and cytotoxic effects in the bivalve mollusks Macoma balthica and Mytilus edulis from the Baltic Sea. Ekologija 54:44–50CrossRefGoogle Scholar
  10. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assay and an assay applicable to acrylamide gels. Anal Biochem 44:276–287CrossRefGoogle Scholar
  11. Bernal-Hernandez YY, Medina-Dıaz IM, Robledo-Marenco ML, Velazquez-Fernandez JB, Giron-Perez MI, Ortega-Cervantes L, Maldonado-Vazquez WA, Rojas-Garcıa AE (2010) Acetylcholinesterase and metallothionein in oysters (Crassostrea corteziensis) from a subtropical Mexican Pacific estuary. Ecotoxicol 19:819–825CrossRefGoogle Scholar
  12. Binelli A, Ricciardi F, Riva C, Provini A (2006) New evidences for old biomarkers: effects of several xenobiotics on EROD and AChE activities in Zebra mussel (Dreissena polymorpha). Chemosphere 62:510–519CrossRefGoogle Scholar
  13. Binelli A, Riva C, Provini A (2007) Biomarkers in Zebra mussel for monitoring and quality assessment of Lake Maggiore (Italy). Biomarkers 12:349–368CrossRefGoogle Scholar
  14. Binelli A, Cogni D, Parolini M, Riva C, Provini A (2009) Cytotoxic and genotoxic effects of in vitro exposure to triclosan and trimethoprim on zebra mussel (Dreissena polymorpha) hemocytes. Comp Biochem Physiol 150C:50–56Google Scholar
  15. Blaise C, Gagne F, Pellerin J, Hansen PD (1999) Determination of vitellogenin-like properties in Mya arenaria hemolymph (Saguenay Fjord, Canada): a potential biomarker for endocrine disruption. Environ Toxicol 14:455–465CrossRefGoogle Scholar
  16. Bonomini M, Dottori S, Amoroso A, Arduini A, Sirolli V (2004) Increased platelet phosphatidylserine exposure and caspase activation in chronic uremia. J Thromb Haemost 2:1275–1281CrossRefGoogle Scholar
  17. Calviello G, Piccioni E, Boninsegna A, Tedesco B, Maggiano N, Serini S, Wolf FI, Palozza P (2006) DNA damage and apoptosis induction by the pesticide mancozeb in rat cells: involvement of the oxidative mechanism. Toxicol Appl Pharmacol 211:87–96CrossRefGoogle Scholar
  18. Cinti DL, Moldeus P, Schenkman JB (1972) Kinetic parameters of drug metabolizing enzymes in Ca2+-sedimented microsomes from rat liver. Biochem Pharmacol 21:3249–3256CrossRefGoogle Scholar
  19. Collin H, Meistertzheim AL, David E, Moraga D, Boutet I (2010) Response of the Pacific oyster Crassostrea gigas, Thunberg 1793, to pesticide exposure under experimental conditions. J Exp Biol 213:4010–4017CrossRefGoogle Scholar
  20. Corsi I, Pastore AM, Lodde A, Palmerini E, Castagnolo L, Focardi S (2007) Potential role of cholinesterases in the invasive capacity of the freshwater bivalve, Anodonta woodiana (Bivalvia: Unionacea): a comparative study with the indigenous species of the genus, Anodonta sp. Comp Biochem Physiol 145(3):413–419Google Scholar
  21. Damiens G, Mouneyrac C, Quiniou F, His E, Gnassia-Barelli M, Roméo M (2006) Metal bioaccumulation and metallothionein concentrations in larvae of Crassostrea gigas. Environ Pollut 140:492–499CrossRefGoogle Scholar
  22. Domico LM, Cooper KR, Bernard LP, Zeevalk GD (2007) Reactive oxygen species generation by the ethylene-bis-dithiocarbamate (EBDC) fungicide mancozeb and its contribution to neuronal toxicity in mesencephalic cells. Neurotoxicology 28:1079–1091CrossRefGoogle Scholar
  23. Domingues I, Agra AR, Monaghan K, Soares AM, Nogueiraa AJ (2010) Cholinesterase and glutathione-S-transferase activities in freshwater invertebrates as biomarkers to assess pesticide contamination. Environ Toxicol Chem 29:5–18CrossRefGoogle Scholar
  24. Ebenso TI, Ita B, Umoren EP, Ita M, Binang W, Edet G, Izah M, Udo IO, Ibanga G, Ukrong EE (2005) Effect of carbamate molluscicide on African giant land snail Lmicolaria aurora. J Appl Sci Environ Manag 9:99–102Google Scholar
  25. Falfushinska HI, Romanchuk LD, Stolyar OB (2008) Different responses of biochemical markers in frogs (Rana ridibunda) from urban and rural wetlands to the effect of carbamate fungicide. Comp Biochem Physiol 148C:223–229Google Scholar
  26. Falfushinska HI, Delahaut L, Stolyar OB, Geffard A, Biagianti-Risbourg S (2009) Multi-biomarkers approach in different organs of Anodonta cygnea from the Dnister basin (Ukraine). Arch Environ Contam Toxicol 57:86–95CrossRefGoogle Scholar
  27. Falfushynska HI, Gnatyshyna LL, Farkas A, Vehovszky A, Gyori J, Stoliar OB (2010) Vulnerability of biomarkers in the indigenous mollusc Anodonta cygnea to spontaneous pollution in a transition country. Chemosphere 81:1342–1351CrossRefGoogle Scholar
  28. Falfushynska HI, Gnatyshyna LL, Golubev AP, Stoliar OB (2012a) Main partitioning criteria for the characterization of the health status in the freshwater mussels Anodonta cygnea from spontaneously polluted area in Western Ukraine. Environ Toxicol 27:485–494CrossRefGoogle Scholar
  29. Falfushynska HI, Gnatyshyna LL, Stoliar OB (2012b) Population-related molecular responses on the effect of pesticides in Carassius auratus gibelio. Comp Biochem Physiol 155C:396–406Google Scholar
  30. Faria M, Carrasco L, Diez S, Riva MC, Bayona JM, Barata C (2009) Multi-biomarker responses in the freshwater mussel Dreissena polymorpha exposed to polychlorobiphenyls and metals. Comp Biochem Physiol 149C:281–288Google Scholar
  31. Figueiredo-Fernandes A, Fontaínhas-Fernandes A, Monteiro R, Reis-Henriques MA, Rocha E (2006) Effects of the fungicide mancozeb on liver structure of Nile tilapia, Oreochromis niloticus: assessment and quantification of induced cytological changes using qualitative histopathology and the stereological point-sampled intercept method. Bull Environ Contam Toxicol 76:249–255CrossRefGoogle Scholar
  32. Gagne F, Blaise C, Hellou J (2004) Endocrine disruption and health effects of caged mussels, Elliptio complanata, placed downstream from a primary-treated municipal effluent plume for 1 year. Comp Biochem Physiol 138C:33–44Google Scholar
  33. Galloway TS, Millward N, Browne MA, Depledge MH (2002) Rapid assessment of organophosphorous/carbamate exposure in the bivalve mollusc Mytilus edulis using combined esterase activities as biomarkers. Aquat Toxicol 61:169–180CrossRefGoogle Scholar
  34. Geracitano LA, Bocchetti R, Monserrat JM, Regoli F, Bianchini A (2004) Oxidative stress responses in two populations of Laeonereis acuta (Polychaeta, Nereididae) after acute and chronic exposure to copper. Mar Environ Res 58:1–17CrossRefGoogle Scholar
  35. Griffith OW (1980) Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem 106:207–212CrossRefGoogle Scholar
  36. Guilhermino L, Barros P, Silva MC, Soares AMVM (1998) Should the use of inhibition of cholinesterases as a specific biomarker for organophosphate and carbamate pesticides be questioned. Biomarkers 3:157–163CrossRefGoogle Scholar
  37. Henderson IF, Triebskorn R (2002) Chemical control in terrestrial gastropods. In: Barker GM (ed) Molluscs as crop pests. CAB International, London, pp 1–31CrossRefGoogle Scholar
  38. Jarrard HE, Delaney KR, Kennedy CJ (2004) Impacts of carbamate pesticides on olfactory neurophysiology and cholinesterase activity in coho salmon (Oncorhynchus kisutch). Aquat Toxicol 69:133–148CrossRefGoogle Scholar
  39. Kádár E, Salánki J, Jugdaohsingh R, Powell JJ, McCrohan CR, Keith N, White KN (2001) Avoidance responses to aluminium in the freshwater bivalve Anodonta cygnea. Aquat Toxicol 55:137–148CrossRefGoogle Scholar
  40. Kang YJ (2006) Metallothionein redox cycle and function. Exp Biol Med 231:1459–1467Google Scholar
  41. Klotz AV, Stegeman JJ, Walsh C (1984) An alternative 7-ethoxyresorufin o-deethylase activity assay: a continuous visible spectrophotometric method for measurement of cytochrome P-450 monooxygenase activity. Anal Biochem 140:138–145CrossRefGoogle Scholar
  42. Kubrak OI, Atamaniuk TM, Husak VV, Drohomyretska IZ, Storey JM, Storey KB, Lushchak VI (2012) Oxidative stress responses in blood and gills of Carassius auratus exposed to the mancozeb-containing carbamate fungicide Tattoo. Ecotoxicol Environ Saf 85:37–43CrossRefGoogle Scholar
  43. Lefcort H, Abbott DP, Cleary DA, Howell E, Keller NC, Smith MM (2004) Aquatic snails from mining sites have evolved to detect and avoid heavy metals. Arch Environ Contam Toxicol 46:478–484CrossRefGoogle Scholar
  44. Lefcort H, Freedman Z, House S, Pendleton M (2008) Hormetic effects of heavy metals in aquatic snails: is a little bit of pollution good? EcoHealth 5:10–17CrossRefGoogle Scholar
  45. Liu J, Shigenaga MK, Yan L-J, Mori A, Ames BN (1996) Antioxidant activity of diethyldithiocarbamate. Free Radical Res 24:461–472CrossRefGoogle Scholar
  46. Lowry OH, Rosebroungh HJ, Farr AL, Randall RJ (1951) Protein measurement with folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  47. Lushchak V, Semchyshyn H, Lushchak O, Mandryk S (2005) Diethyldithiocarbamate inhibits in vivo Cu, Zn-superoxide dismutase and perturbs free radical processes in the yeast Saccharomyces cerevisiae cells. Biochem Biophys Res Commun 338:1739–1744CrossRefGoogle Scholar
  48. Lushchak VI, Bagnyukova TV, Lushchak OV, Storey JM, Storey KB (2007) Diethyldithiocarbamate injection induces transient oxidative stress in goldfish tissues. Chem Biol Interact 170:1–8CrossRefGoogle Scholar
  49. Marchi B, Burlando B, Moore MN, Viarengo A (2004) Mercury- and copper-induced lysosomal membrane destabilization depends on [Ca2+]idependent phospholipase A2 activation. Aquat Toxicol 66:197–204CrossRefGoogle Scholar
  50. Mdegela RH, Mosha RD, Sandvik M, Skaare JU (2010) Assessment of acetylcholinesterase activity in Clarias gariepinus as a biomarker of organophosphate and carbamate exposure. Ecotoxicology 19:855–863CrossRefGoogle Scholar
  51. Mora P, Fournier D, Narbonne JF (1999) Cholinesterases from the marine mussels Mytilus galloprovincialis Lmk. and M. edulis L. and from the freshwater bivalve Corbicula fluminea Muller. Comp Biochem Physiol 122(3):353–361Google Scholar
  52. Nielson KB, Winge DR (1984) Preferential binding of copper to the beta domain of metallothionein. J Biol Chem 259:4941–4946Google Scholar
  53. Noel S, Billo Bah B (2009) Risks assessment of water pollution by pesticides at local scale (PESTEAUX project): study of polluting pressure. Commun Agric Appl Biol Sci 74:165–170Google Scholar
  54. Ohkawa H, Ohishi N, Tagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358CrossRefGoogle Scholar
  55. Olive PL (1988) DNA precipitation assay: a rapid and simple method for detecting DNA damage in mammalian cells. Environ Mol Mutagen 11:487–495CrossRefGoogle Scholar
  56. Reznick AZ, Packer L (1994) Oxidative damage to proteins: spectrophotometric method for carbonyl assay. Meth Enzymol 233:357–363CrossRefGoogle Scholar
  57. Rittschof D, McClellan-Green P (2005) Molluscs as multidisciplinary models in environment toxicology. Mar Pollut Bull 50:369–373CrossRefGoogle Scholar
  58. Robillard S, Beauchamp G, Laulier M (2003) The role of abiotic factors and pesticide levels on enzymatic activity in the freshwater mussel Anodonta cygnea at three different exposure sites. Comp Biochem Physiol 135C:49–59Google Scholar
  59. Roesijadi G, Fowler B (1991) Purification of invertebrate metallothioneins. Methods Enzymol 205B:263–273CrossRefGoogle Scholar
  60. Romero A, Estevez-Calvar N, Dios S, Figueras A, Novoa B (2011) New insights into the apoptotic process in mollusks: characterization of caspase genes in Mytilus galloprovincialis. PLoS ONE 6(2):e17003CrossRefGoogle Scholar
  61. Rotter S, Sans-Piché F, Streck G, Altenburger R, Schmitt-Jansen M (2011) Active bio-monitoring of contamination in aquatic systems—an in situ translocation experiment applying the PICT concept. Aquat Toxicol 101:228–236CrossRefGoogle Scholar
  62. Shi D, Wang WX (2004) Understanding the differences in Cd and Zn bioaccumulation and subcellular storage among different populations of marine clams. Environ Sci Technol 38:449–456CrossRefGoogle Scholar
  63. Sole M, Porte C, Albaigh J (1995) The use of biomarkers for assessing the effects of organic pollution in mussels. Sci Total Environ 159:147–153CrossRefGoogle Scholar
  64. Srivastava AK, Ali W, Singh R, Bhui K, Tyagi S, Al-Khedhairy AA, Srivastava PK, Musarrat J, Shukla Y (2012) Mancozeb-induced genotoxicity and apoptosis in cultured human lymphocytes. Life Sci 90:815–824CrossRefGoogle Scholar
  65. Viarengo A, Ponzano E, Dondero F, Fabbri R (1997) A simple spectrophotometric method for metallothionein evaluation in marine organisms: an application to Mediterranean and Antarctic molluscs. Mar Environ Res 44:69–84CrossRefGoogle Scholar
  66. Viarengo A, Burlando B, Dondero F (1999) Metallothionein as a tool in biomonitoring programmes. Biomarkers 4:455–466CrossRefGoogle Scholar
  67. Viarengo A, Lowe D, Bolognesi C, Fabbri E, Koehler A (2007) The use of biomarkers in biomonitoring: a 2-tier approach assessing the level of pollutant-induced stress syndrome in sentinel organisms. Comp Biochem Physiol 146C:281–300Google Scholar
  68. Wang WX, Rainbow P (2008) Comparative approaches to understand metal bioaccumulation in aquatic animals. Comp Biochem Physiol 148C:315–323Google Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Halina I. Falfushynska
    • 1
  • Lesya L. Gnatyshyna
    • 1
  • Oksana B. Stoliar
    • 1
    Email author
  1. 1.Research Laboratory of Comparative Biochemistry and Molecular BiologyTernopil National Pedagogical UniversityTernopilUkraine

Personalised recommendations