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Multiple Biomarker Comparison in Mytilus galloprovincialis from the Greece Coast: “Lysosomal Membrane Stability, Neutral Red Retention, Micronucleus Frequency and Stress on Stress”

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Abstract

The lysosomal membrane stability test applied on the digestive cells (LMS) and the neutral red lysosomal retention assay (NRR) performed on hemocytes have been evaluated on mussels Mytilus galloprovincialis collected from Thermaikos and Strymonikos gulfs (Nothern Greece) in June and December 2000. The correlation analysis indicated that there was a strong positive relationship between the above mentioned biomarkers in both sampling seasons, supporting the validity of their application in water quality assessment. The evaluation of micronucleus frequency (MN) has been also performed in hemocytes of the same mussels and, according to the results, it requires additional validation before using it as a stress index. The results obtained in parallel by the stress on stress (SOS) technique do not support the application of this biomarker in biomonitoring, showing, however, negative correlation with LMS and NRR in one sampling season. Finally, morphological observations were performed on cryosections stained for the enzyme N-acetyl-β-hexosaminidase, revealing differences in the epithelial cell-layer thickness, as well as changes in the digestive lysosomal system of mussels, obtained from different sampling sites in the two sampling seasons.

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References

  • T.A. Albanis T.G. Danis M.K Kourgia (1994) ArticleTitleTransportation of pesticides in estuaries of Axios, Loudias and Aliakmon rivers (Thermaikos Gulf), Greece Sci. Total Environ. 156 11–22

    Google Scholar 

  • Anagnostou, C., Georgakopoulou-Gregoriadou, E., Zenetos, A., Kamberi E., Karageorgis, A., Kontogiannis, C., Pagou, K., Panagiotidis, P., Pancucci, A., Siokou, I., Symbura, N., Chathianestis, I. and Psyllidou-Giuranovits, R. (1997). The state of the marine ecosystem of the bay and the Gulf of Thessaloniki – The reaction capacity of the ecosystem. Proceedings of the 5th Panellenic Symposium on Oceanography and Fisheries, Kavala, Greece, 15–18/04/97, 1, 83–6

  • C. Anagnostou H. Kamberi A Karageorgis (1998) ArticleTitleHorizontal and vertical distribution in heavy metals in sediments from Thermaikos Gulf Rapp Comm. Int. Mer. Medit. 35 222

    Google Scholar 

  • K. Becker-van Slooten J Tarradellas (1995) ArticleTitleOrganotins in Swiss lakes after their ban: assessment of water, sediment, and Dreissena polymorpha contamination over a four-year period Arch. Environ. Contam. Toxicol. 29 284–392

    Google Scholar 

  • C. Bolognesi E. Landini P. Roggieri R. Fabbri A Viarengo (1999) ArticleTitleGenotoxicity biomarkers in the assessment of heavy metal effects in mussels: experimental studies Environ. Mol. Mutagen. 33 IssueID4 287–92

    Google Scholar 

  • R. Brunetti F. Majone I. Gola C Beltrame (1988) ArticleTitleThe micronucleus test: examples of application to marine ecology Mar. Ecol. Prog. Ser. 44 65–8

    Google Scholar 

  • M.P. Cajaraville J.A. Marigomez E.A Angulo (1989) ArticleTitleA stereological survey of lysosomal structure alterations in Littorina littorea exposed to 1-naphtol Comp. Biochem. Physiol. 93C 231–37

    Google Scholar 

  • M.P. Cajaraville J.A. Marigomez G. Diez E.A Angulo (1992) ArticleTitleComparative effects of the water accommodated fraction of three oils on mussels 2. Quantitative alterations in the structure of the digestive tubules Comp. Biochem. Physiol. 102C 113–23

    Google Scholar 

  • M. Camusso R. Balestrini F. Muriano M Mariani (1994) ArticleTitleUse of freshwater mussel Dreissena polymorpha to assess trace metal pollution in the lower river Po (Italy) Chemosphere. 29 729–45

    Google Scholar 

  • Catsiki, V., Hatzianestis, I., Strogyloudi, E., Belou, O., Gogou, M. and Rigas, F. (2001) Distribution of metals and organic contaminants in mussels from Thermaikos gulf. Proceedings of the 7th International Conference on Environmental Science and Technology. Syros island, Greece, 3–7/09/01, Vol A, pp. 106–13

  • S. Dailianis G.P. Domouhtsidou E. Raftopoulou M. Kaloyanni V.K Dimitriadis (2003) ArticleTitleEvaluation of neutral red retention assay, micronucleus test, acetylcholinesterase activity and a signal transduction molecule (cAMP) in tissues of Mytilus galloprovincialis (L.), in pollution monitoring Mar. Environ. Res. 56 443–70

    Google Scholar 

  • R.K. Das N.K Nanda (1986) ArticleTitleInduction of micronuclei in peripheral erythrocytes of fish Heteropneustes fossilis by mitomycin C and paper mill effluent Mutat. Res. 175 67–71

    Google Scholar 

  • G.P. Domouhtsidou V.K Dimitriadis (2001) ArticleTitleLysosomal and lipid alterations in the digestive gland of mussels, Mytilus galloprovincialis (L.) as biomarkers of environmental stress Environ. Poll. 115 123–37

    Google Scholar 

  • M. Etxeberria M.P. Cajaraville I Marigomez (1995) ArticleTitleChanges in digestive cell lysosomal structure in mussels as biomarkers of environmental stress in the Urdaibai estuary (Biscay Coast, Iberian Peninsula) Mar. Poll. Bull. 9 IssueID30 599–603

    Google Scholar 

  • M. Etxeberria I. Sastre M.P. Cajaraville I Marigomez (1994) ArticleTitleDigestive lysosome enlargement induced by experimental exposure to metals (Cu, Cd and Zn) in mussels collected from a Zn-polluted site Arch. Environ. Contam. Toxicol. 27 338–45

    Google Scholar 

  • M. Fenech W.P. Chang M. Kirsch-Volders N. Holland S. Bonassi E Zeiger (2003) ArticleTitleHUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures Mutat. Res. 534 65–75

    Google Scholar 

  • P.W. Fernley M.N. Moore D.M. Lowe P. Donkin S Evans (2000) ArticleTitleImpact of the Sea Empress oil spill on lysosomal stability in mussel blood cells Mar. Environ. Res. 50 451–55

    Google Scholar 

  • F.L. Harrison R Berger (1982) ArticleTitleEffects of copper on the latency of lysosomal hexosaminidase in the digestive cells of Mytilus edulis Mar. Biol. 68 109–16

    Google Scholar 

  • I. Hatzianestis E. Sklivagou E Georgakopoulou (2000) ArticleTitleOrganic contaminants in sediments and mussels from Thermaikos Gulf, Hellas Toxicol. Environ. Chem. 74 203–16

    Google Scholar 

  • J.A. Heddle M.C. Cimino M. Hayashi F. Romagna M.D. Shelby J.D. Tucker Vanparys Ph. J.T Macgregor (1991) ArticleTitleMicronuclei as an index of cytogenetic damage: past, present, and future Environ. Mol. Mutagen. 18 277–91

    Google Scholar 

  • A. Jaylet P. Deparis V. Ferrier S. Grinfeld R Siboulet (1986) ArticleTitleA new micronucleus test using peripheral blood erythrocytes of the newt Pleurodeles waltl to detect mutagens in fresh-water pollution Mutat. Res. 164 245–57

    Google Scholar 

  • Kamarianos, A., Karamanlis, X. and Galoupi, E. (2002). Pollution of coastal areas of N. Greece by organochlorine pesticides and polychlorinated biphenyls (PCBs). Proceedings of the 1st Conference of Macedonia, Thessaloniki, Greece, 1–4/03/02, pp 116–21

  • Kamarianos, A. and Kilikidis, S. (1997). Pollution monitoring of Thermaikos Gulf using organochloride pesticides and polychlorinated biphenyls (PCBs), (1991–1996). Proceedings of the 5th Conference of Environmental Science and Technology. Lesvos island, Greece. 1–4/09/97, Vol B, pp. 326–33

  • P.K. Krishnakumar E. Cassillas U Varanasi (1994) ArticleTitleEffect of environmental contamination on the health of Mytilus edulis from Puget Sound, Washington: cytochemical measures of lysosomal responses in the digestive cells using automatic image analysis Mar. Ecol. Prog. Ser. 106 249–61

    Google Scholar 

  • D.M. Lowe K.R Clarke (1989) ArticleTitleContaminant induced changes in the structure of the digestive epithelium of Mytilus edulis Aquat. Toxicol. 15 345–58

    Google Scholar 

  • D.M. Lowe R.K Pipe (1994) ArticleTitleContaminant induced lysosomal membrane damage in marine mussel digestive cells: an in vitro study Aquat. Toxicol. 30 357–65

    Google Scholar 

  • D.M. Lowe V.U. Fossato M.H Depledge (1995a) ArticleTitleContaminant-induced lysosomal membrane damage in blood cells of mussels Mytilus galloprovincialis from the Venice Lagoon: an in vitro study Mar. Ecol. Prog. Ser. 129 189–96

    Google Scholar 

  • D.M. Lowe C. Soverchia M.N Moore (1995b) ArticleTitleLysosomal membrane responses in the blood and digestive cells of mussels experimentally exposed to fluoranthene Aquat. Toxicol. 33 105–12

    Google Scholar 

  • F. Majone R. Brunetti I. Gola A.G Levis (1987) ArticleTitlePersistence of micronuclei in the marine mussel, Mytilus galloprovincialis, after treatment with mitomycin C Mutat. Res. 191 157–61

    Google Scholar 

  • I. Marigomez A. Orbea I. Olabarrieta M. Exteberria M.P Cajaraville (1996) ArticleTitleStructural changes in the digestive lysosomal system of sentinel mussels as biomarkers of environmental stress in mussel-watch programmes Comp. Biochem. Physiol. 113C IssueID2 291–7

    Google Scholar 

  • J. Mersch M.N Beauvais (1997) ArticleTitleThe micronucleus assay in the zebra mussel, Dreissena polymorpha, to in situ monitor genotoxicity in freshwater environments Mutat. Res. 393 141–9

    Google Scholar 

  • J. Mersch M.N. Beauvais P Nagel (1996) ArticleTitleInduction of micronuclei in haemocytes and gill cells of zebra mussel, Dreissena polymorpha, exposed to clastogens Mutat. Res. 371 47–55

    Google Scholar 

  • M.N Moore (1976) ArticleTitleCytochemical demonstration of latency of lysosomal hydrolases in digestive cells of the common mussel Mytilus edulis, and changes induced by thermal stress Cell Tissue Res. 175 IssueID3 279–87

    Google Scholar 

  • G. Nikolaidis M Moustaka-Gouni (1990) ArticleTitleThe structure and dynamics of phytoplankton assemblages from the inner part of the Thermaikos Gulf, Greece. I. Phytoplankton composition and biomass from May 1988 to April 1989 Helgolander Meeresun. 44 487–501

    Google Scholar 

  • F. Regoli E Orlando (1993) ArticleTitleMytilus galloprovincialis as a bioindicator of lead pollution: biological variables and cellular responses Sci. Tot. Environ. Suppl 1283–92

    Google Scholar 

  • A.H. Ringwood D.E. Conners J Hoguet (1998) ArticleTitleEffects of natural and anthropogenic stressors on lysosomal destabilization in oysters Crassostrea virginica Mar. Ecol. Prog. Ser. 166 163–71

    Google Scholar 

  • R. Scarpato L. Migliore G. Alfinito-Cognetti R Barale (1990) ArticleTitleInduction of micronuclei in gill tissue of Mytilus galloprovincialis exposed to polluted marine waters Mar. Pollut. Bull. 21 IssueID2 74–80

    Google Scholar 

  • A.C. Smaal A. Wagenvoort J. Hemelraad I Akkerman (1991) ArticleTitleResponse to stress of mussels (Mytilus edulis) exposed in Dutch tidal waters Comp. Biochem. Physiol. C 100 IssueID1–2 197–200

    Google Scholar 

  • X. Stien P. Percic M. Gnassia-Barelli M. Rome M Lafaurie (1998) ArticleTitleEvaluation of biomarkers in caged fishes and mussels to assess the quality of waters in a bay of the NW Mediterranean Sea Environ. Pollut. 99 3 339–45

    Google Scholar 

  • H. Stopper S.O Muller (1997) ArticleTitleMicronuclei as a biological endpoint for genotoxicity: a minireview Toxicol. In Vitro 11 661–7

    Google Scholar 

  • C. Svendsen J.M Weeks (1995) ArticleTitleThe use of a lysosome assay for the rapid assessment of cellular stress from copper to the freshwater snail Viviparus contectus (Millet) Mar. Pollut. Bull. 31 139–42

    Google Scholar 

  • R.E. Thomas P.M. Harris R.D Rice (1999) ArticleTitleSurvival in air of Mytilus trossulus following long-term exposure to spilled Exxon Valdez crude oil in Prince William sound Comp. Biochem. Physiol. C: Comp. Pharm. Toxicol. 122 IssueID1 147–52

    Google Scholar 

  • R. Tremblay B. Myrand H Guderley (1998) ArticleTitleTemporal variation of lysosomal capacities in relation to susceptibility of mussels, Mytilus edulis, to summer mortality Mar. Biol. 132 IssueID4 641–9

    Google Scholar 

  • UNEP, (1997). Intercomparison Exercise Concerning Lysosomal Membrane Stability and Metallothionein Analysis. UNEP(OCA)/MED WG. 132/5. Athens, Greece. pp.1–12

  • UNEP/RAMOGE, (1999). Manual on the Biomarkers Recommended for the MED POL Biomonitoring Programme, UNEP, Athens, Greece. pp. 1–92

  • M.A. Gaag ParticleVan der A. Gauthier Y. Noordsij Levi M.N. Wrisberg (1990) Methods to measure genotoxins in wastewater: evaluation with in vivo and in vitro tests M.D. Waters (Eds)   EditionNumber6 Plenum Press New York 215–32

    Google Scholar 

  • M.M. Vega J.A. Marigomez E Angulo (1989) ArticleTitleQuantitative alterations in digestive cell structure of the marine gastropod Littorina littorea exposed to cadmium Mar. Biol. 103 547–53

    Google Scholar 

  • M.B. Veldhuizen-Tsoerkan D.A. Holwerda A..M. Bont Particlede A.C. Smaal D.I Zandee (1991) ArticleTitleA field study on stress indices in the sea mussel, Mytilus edulis: application of the “stress approach” in biomonitoring Arch. Environ. Contam. Toxicol. 21 IssueID4 497–504

    Google Scholar 

  • A. Viarengo L. Canesi M. Pertica G. Mancinelli R. Accomando A.C. Smaal M Orunesu (1995) ArticleTitle“Stress on stress” response: a simple monitoring tool in the assessment of a general stress syndrome in mussels Mar. Environ. Res. 39 245–8

    Google Scholar 

  • A. Viarengo M.N. Moore G. Mancinelli A. Mazzucotelli R.K. Pipe S.V Farrar (1987) ArticleTitleMetallothioneins and lysosomes in metal toxicity and accumulation in marine mussels: the effect of cadmium in the presence and absence of phenanthrene Mar. Biol. 94 251–7

    Google Scholar 

  • Voutsinou-Taliadouri, F., Hatzianestis, I. and Georgakopoulou-Gregoriadou, E. (1998). Trace elements, pesticides and PCBs levels in sediments of a bay influenced by anthropogenic activities (Thermaikos bay, N.W. Aegean Sea). International Symposium on Marine Pollution, Extended synopses, Monaco, 5–9/10/98, 668–80

  • M.N. Wrisberg C.M. Bilbo H Spliid (1992) ArticleTitleInduction of micronuclei in hemocytes of Mytilus edulis and statistical analysis Ecotoxicol. Environ. Saf. 23 191–205

    Google Scholar 

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Koukouzika, N., Dimitriadis, V.K. Multiple Biomarker Comparison in Mytilus galloprovincialis from the Greece Coast: “Lysosomal Membrane Stability, Neutral Red Retention, Micronucleus Frequency and Stress on Stress”. Ecotoxicology 14, 449–463 (2005). https://doi.org/10.1007/s10646-004-1350-9

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