Marine Biology

, Volume 149, Issue 1, pp 87–96 | Cite as

Swimming speed alteration of larvae of Balanus Amphitrite as a behavioural end-point for laboratory toxicological bioassays

  • M. FaimaliEmail author
  • F. Garaventa
  • V. Piazza
  • G. Greco
  • C. Corrà
  • F. Magillo
  • M. Pittore
  • E. Giacco
  • L. Gallus
  • C. Falugi
  • G. Tagliafierro
Research Article


In this study, we investigate the feasibility of developing a new behavioural toxicity bioassay (Swimming Speed Alteration test—SSA test) with larvae of Balanus amphitrite (Crustacea Cirripedia). This organism was chosen as a model for different reasons: it is present all over the world, simple to be reared, easily available, and also because barnacles play an important role in the coastal ecosystem. In addition, all the operations related to the rearing and test execution are comparatively cheap. This bioassay was performed with several classes of chemical pollutants (antifouling biocides, neurotoxic pesticides, and heavy metals) and with environmental samples (sediment elutriates). The measurement of swimming speed, by means of video-graphic techniques, proved to be a valid instrument in highlighting the sub-lethal levels of toxicity caused by the different tested samples. In conclusion, the SSA test is able to provide in a biomonitoring program a good behavioural integrated output, which is also repeatable, sensitive, easily interpretable, and truly representative of a broad range of toxic compounds and environmental toxic matrices which are, generally, very complex and difficult to analyse. For all of these reasons, it could be proposed as a non-specific behavioural end-point.


Swimming Speed Swimming Behaviour Methomyl Stage Nauplius Pyrithione 
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 work was partially supported by MIUR funds, Project MEMOBIOMAR (law 248).


  1. ASTM (1992) Designation E 1367: standard guide for conducting 10-day static sediment toxicity tests with marine and estuarine amphipods, vol 11.04. American Society for Testing and Materials, Philadelphia, PAGoogle Scholar
  2. Baillieul M, Blust R (1999) Analysis of the swimming velocity of cadmium-stressed Daphnia magna. Aquat Toxicol 44:245–254CrossRefGoogle Scholar
  3. Bat L, Raffaelli D, Marr IL (1998) The accumulation of copper, zinc and cadmium by the amphipod Corophium volutator (Pallas). J Exp Mar Biol Ecol 223(2):167–184CrossRefGoogle Scholar
  4. Bayley M, Baatrup E (1996) Pesticide uptake and locomotor behaviour in the woodlouse: an experimental study employing video-tracking and 14C-labelling. Ecotoxicology 5:35–45CrossRefGoogle Scholar
  5. Beauvais SL, Jones SB, Brewer SK, Little EE (2000) Physiological measure of neurotoxicity of diazinon and malathion to larval rainbow trout (Oncorhynchus mykiss) and their correlation with behavioural measures. Environ Toxicol Chem 19(7):1875–1880CrossRefGoogle Scholar
  6. Beiras R, Bellas J, Fernandez N, Lorenzo JI, Cobelo-Garcia A (2003) Assessment of coastal marine pollution in Galicia (NW Iberian Peninsula); metal concentrations in seawater, sediments and mussels (Mytilus galloprovincialis) versus embryo-larval bioassays using Paracentrotus lividus and Ciona intestinalis. Mar Environ Res 56(4):531–553CrossRefGoogle Scholar
  7. Beiras R, Vazquez E, Bellas J, Lorenzo JI, Fernandez N, Macho G, Marino JC, Casas L (2001) Sea-urchin embryo bioassay for in situ evaluation of the biological quality of coastal seawater. Estuar Coast Shelf Sci 52:29–32CrossRefGoogle Scholar
  8. Boenigk J, Wiedlroither A, Pfandl K (2005) Heavy metal toxicity and bioavailability of dissolved nutrients to a bacterivorous flagellate are linked to suspended particle physical properties. Aquat Toxicol 71(3):249–259CrossRefGoogle Scholar
  9. Borgmann U (2000) Methods for assessing the toxicological significance of metals in aquatic ecosystems: bio-accumulation-toxicity relationships, water concentrations and sediment spiking approaches. Aquat Ecosyst Health Manag 3:277–289Google Scholar
  10. Briggs AD, Greenwood N, Grant A (2003) Can turbidity caused by Corophium volutator (Pallas) activity be used to assess sediment toxicity rapidly? Mar Environ Res 55:181–192CrossRefGoogle Scholar
  11. Byrne PA, O’Halloran J (1999) Aspects of assaying sediment toxicity in Irish estuarine ecosystems. Mar Poll Bull 39(1–12):97–105CrossRefGoogle Scholar
  12. Byrne PA, O’Halloran J (2000) Acute and sublethal toxicity of estuarine sediments to the manila clam, Tapes semidecussatus. Environ Toxicol 15(5):456–468CrossRefGoogle Scholar
  13. Calabrese EJ (2002) Hormesis: changing view of the dose-response, a personal account of the history and current status. Mutat Res 511(3):181–189CrossRefGoogle Scholar
  14. Calabrese EJ, Baldwin LA (2001a) Hormesis: u-shaped dose responses and their centrality in toxicology. Trends Pharmacol Sci 22(6):285–291CrossRefGoogle Scholar
  15. Calabrese EJ, Baldwin LA (2001b) The frequency of u-shaped dose responses in the toxicological literature. Toxicol Sci 62:330–338CrossRefGoogle Scholar
  16. Charoy CP, Janssen CR, Persoone G, Clément P (1995) The swimming behaviour of Brachionus calyciflorus (rotifer) under toxic stress. I. The use of automated trajectometry for determining sublethal effects of chemicals. Aquat Toxicol 32:271–282CrossRefGoogle Scholar
  17. Charoy CP, Janssen CR (1999) The swimming behaviour of Brachionus calyciflorus (rotifer) under toxic stress. II. Comparative sensitivity of various behavioural criteria. Chemosphere 38:3247–3260CrossRefGoogle Scholar
  18. Cheung KC, Wong MH, Yung YK (2003) Toxicity assessment of sediments containing tributyltin around Hong Kong harbour. Toxicol Lett 137(1–2):121–131CrossRefGoogle Scholar
  19. Cohn SA, McGuire JR (2000) Using diatom motility as an indicator of environmental stress: effects of toxic sediment elutriates. Diatom Res 15(1):19–29CrossRefGoogle Scholar
  20. Depledge MH, Aagaard A, Györkös P (1995) Assessment of trace metal toxicity using molecular, physiological and behavioural biomarkers. Mar Poll Bull 31:19–27CrossRefGoogle Scholar
  21. Environment Canada (1994) Collection and preparation of sediment for physicochemical characterization and biological testing. EPS 1/RM/29Google Scholar
  22. EPA (2001) Methods for collection, storage and manipulation of sediments for chemical and toxicological analyses: technical manual. United States environmental protection office of water 4305, EPA-823-F-01-023Google Scholar
  23. Faimali M, Magillo F, Piazza V, Garaventa F, Geraci S (2002) A simple toxicological bioassay using phototactic behaviour of Balanus amphitrite (Darwin) nauplii: role of some cultural parameters and application with experimental biocides. Period Biol 104(2):217–223Google Scholar
  24. Faimali M, Falugi C, Gallus L, Piazza V, Tagliaferro G (2003a) Involment of acetyl choline in settlement of Balanus amphitrite. Biofouling 19(Suppl):213–220CrossRefGoogle Scholar
  25. Faimali M, Sepčić K, Turk T, Geraci S (2003b) Non-toxic antifouling–activity of polymeric 3-alkylpyridinium salts from the Mediterranean sponge Reniera sarai (Pulitzer-Finali). Biofouling 19(1):47–56CrossRefGoogle Scholar
  26. Falugi C (1988) Localisation and possible functions of cholinesterase activities in Balanus amphitrite embryos and larvae. Acta Embryol Morphol Exp NS 9:133–156Google Scholar
  27. Farr JA (1977) Impairment of antipredator behaviour in Palaemonetes pugio by exposure to sub-lethal doses of parathion. Trans Am Fish Soc 106:287–290CrossRefGoogle Scholar
  28. Finney DJ (1978) Statistical method in biological assay, 3rd edn. Charles Griffin & Co. Ltd, London, England, p 508Google Scholar
  29. Gaudy R, Guérin JP, Kerambrun P (1991) Sublethal effects of cadmium on respiratory metabolism, nutrition, excretion and hydrolyse activity in Leptomysis lingvura (Crustacea: Mysidacea). Mar Biol 109:493–501CrossRefGoogle Scholar
  30. Giesy JP, Rosiu CJ, Graney RL, Henry MG (1990) Benthic invertebrate bioassays with toxic sediment and porewater. Environ Toxicol Chem 9:233–248CrossRefGoogle Scholar
  31. Geffard A, Geffard O, His E, Amiard JC (2002) Relationships between metal bioaccumulation and metallothionein levels in larvae of Mytilus galloprovincialis exposed to contaminated estuarine sediment elutriate. Mar Ecol Prog Ser 233:131–142CrossRefGoogle Scholar
  32. Geffard O, Budzinski H, LeMenach K (2004) Chemical and ecotoxicological characterization of the “Erika” petroleum: bio-tests applied to petroleum water-accommodated fractions and natural contaminated samples. Aquat Living Resour 17(3):289–296CrossRefGoogle Scholar
  33. Geffard O, Geffard A, His E, Budzinski H (2003) Assessment of the bioavailability and toxicity of sediment-associated polycyclic aromatic hydrocarbons and heavy metals applied to Crassostrea gigas embryos and larvae. Mar Poll Bull 46:481–490CrossRefGoogle Scholar
  34. Goka K (1999) Enbryotoxicity of zinc pyrithione, an antidandruff chemical, in fish. Environ Res 81:81–83CrossRefGoogle Scholar
  35. Goto T, Hiromi J (2003) Toxicity of 17α-ethynylestradiol and norethindrone, constituents of an oral contraceptive pill to the swimming and reproduction of cladoceran Daphnia magna, with special reference to their synergetic effect. Mar Poll Bull 47:139–142CrossRefGoogle Scholar
  36. Häder DP, Lebert M (1985) Real-time computer-controlled tracking of motile microorganisms. Photochem Photobiol 42:509–514CrossRefGoogle Scholar
  37. His E, Beiras R, Seaman MNL (1999) The assessment of aquatic contamination: bioassays with bivalve embryos and larvae. Adv Mar Biol 37:1–178CrossRefGoogle Scholar
  38. Hopkins WA, Snodgrass JW, Staub BP, Jackson BP, Congdon JD (2003) Altered swimming performance of a benthic fish (Erimyzon sucetta) exposed to contaminated sediments. Arch Environ Contam Toxicol 44:383–389CrossRefGoogle Scholar
  39. Kane AS, Salierno JD, Gipson GT, Molteno TCA, Hunter C (2004) A video-based movement analysis system to quantify behavioural stress responses of fish. Water Res 38:3993–4001CrossRefGoogle Scholar
  40. Kater BJ, Postma JF, Dubbeldam M, Prins JTHJ (2001) Comparison of laboratory and in situ sediment bioassays using Corophium volutator. Environ Toxicol Chem 20(6):1291–1295CrossRefGoogle Scholar
  41. Lam PKS, Wo KT, Wu RSS (2000) Effects of cadmium on the development and swimming behaviour of barnacle larvae Balanus amphitrite Darwin. Environ Toxicol 15(1):8–13CrossRefGoogle Scholar
  42. Lang WH, Forward RB, Miller DC, Marcy M (1980) Acute toxicity and sublethal behavioural effects of copper on barnacle nauplii. Mar Biol 58:139–145CrossRefGoogle Scholar
  43. Lang WH, Miller DC, Ritacco PJ, Marcy M (1981) The effects of copper and cadmium on the behaviour and development of barnacle larvae. In: Vernberg J, Calabrese A, Thurburg FP, Vernberg WB (eds) Biological monitoring of marine pollutants. Academic Press, New York, pp 165–203CrossRefGoogle Scholar
  44. Little EE, Archeski RD, Flerov BA, Kozlovskaya VI (1990) Behavioral indicators of sublethal toxicity in rainbow trout. Arch Environ Contam Toxicol 19:380–385CrossRefGoogle Scholar
  45. Little EE, Finger SE (1990) Swimming behaviour as an indicator of sublethal toxicity in fish. Environ Toxicol Chem 9:13–19CrossRefGoogle Scholar
  46. Losso C, Novelli AA, Picone M, Marchetto D, Pessa G, Molinaroli E, Ghetti R, Ghirardini AV (2004) Evaluation of surficial sediment toxicity and sediment physico-chemical characteristics of representative sites in the Lagoon of Venice (Italy). J Mar Syst 51(1–4):281–292CrossRefGoogle Scholar
  47. Mackey DJ, Butler KCV, Carpenter PD, Higgins HW, O’Sullivan JE, Plaschke RB (1996) Trace elements and organic matter in a pristine environment: bathurst Harbor, southwestern Tasmania. Sci Total Environ 191:137–151CrossRefGoogle Scholar
  48. Madsen T, Gustavson K, Samsøe-Petersen L, Simonsen F, Jakobsen J, Foverskov S, Larsen MM (2000) Ecotoxicological assessments of antifouling biocides and nonbiocidal paints. Environmental Project, No 531. Danish Environmental Protection AgencyGoogle Scholar
  49. Magillo F, Faimali M, Andrenacci M, Geraci S (2002) Video-track analysis of light-induced motion response in Balanus amphitrite larvae. Biol Mar Medit 9(1):852–855Google Scholar
  50. Magillo F, Faimali M, Geraci S (2003) Effect of cadmium chloride on the swimming behaviour of Balanus amphitrite (Crustacea: cirripedia) larvae. Biol Mar Med 10(2):1014–1017Google Scholar
  51. Maraldo K, Dahllöf I (2004) Seasonal variations in the effect of zinc pyrithione and copper pyrithione on pelagic phytoplankton communities. Aquat Toxicol 69:189–198CrossRefGoogle Scholar
  52. Mueller DC, Bonner JS, McDonald SJ, Autenrieth RL, Donnelly KC, Lee K, Doe K, Anderson J (2003) The use of toxicity bioassays to monitor the recovery of oiled wetland sediments. Environ Toxicol Chem 22(9):1945–1955CrossRefGoogle Scholar
  53. Nascimento A, Smith DH, Pereira SA, Sampaio de Araujo MM, Silva MA, Mariani AM (2000) Integration of varying responses of different organisms to water and sediment quality at sites impacted and not impacted by the petroleum industry. Aquat Ecosys Health Manag 3:449–458CrossRefGoogle Scholar
  54. Norris DO, Donahue S, Dores RM, Lee JK, Maldonado TA, Ruth T, Woodling JD (1999) Impaired adrenocortical response to stress by brown trout, Salmo trutta, living in metal-contaminated waters of the Eagle River, Colorado. Gen Comp Endocrinol 113:1–8CrossRefGoogle Scholar
  55. Okamura H, Watanabe T, Aoyama I, Hasobe M (2002) Toxicity evaluation of new antifouling compounds using suspension-cultured fish cells. Chemosphere 46:945–951CrossRefGoogle Scholar
  56. Onorati F, Mecozzi M (2004) Effects of two diluents in the Microtox® toxicity bioassay with marine sediments. Chemosphere 54(5):679–687CrossRefGoogle Scholar
  57. Pelosi S, Franchi M (2003) Multiple survey of environmental conditions in Varano lagoon (south Italy). Quim Nova 26(6):789–794CrossRefGoogle Scholar
  58. Petrauskiene L (2003) Water and sediment toxicity assessment by use of behavioural responses of medicinal leeches. Environ Int 28:729–736CrossRefGoogle Scholar
  59. Piazza V, Faimali M, Geraci S (2003) Settlement of Balanus amphitrite as a possible behavioural parameter in an alternative toxicological assay. Biol Mar Medit 10(2):1115–1118Google Scholar
  60. Power M, Attrill MJ, Thomas RM (1999) Trends in agricultural pesticide (atrazine, lindane, simazine) concentrations in the Thames Estuary. Environ Pollut 104:31–39CrossRefGoogle Scholar
  61. Raven PJ, George JJ (1989) Recovery by riffle macroinvertebrates in a river after a major accidental spillage of chlorpyrifos. Environ Pollut 59:55–70CrossRefGoogle Scholar
  62. Roast SD, Widdows J, Jones MB (2000a) Mysids and trace metals: disruption of swimming as a behavioural indicator of environmental contamination. Mar Environ Res 50:107–112CrossRefGoogle Scholar
  63. Roast SD, Widdows J, Jones MB (2000b) Disruption of swimming in the hyperbenthic mysid Neomysis integer (Peracarida: Mysidacea) by the organophosphate pesticide chlorpyrifos. Aquat Toxicol 47:227–241CrossRefGoogle Scholar
  64. Roast SD, Widdows J, Jones MB (2001) Impairment of mysid (Neomysis integer) swimming ability: an environmentally realistic assessment of the impact of cadmium exposure. Aquat Toxicol 52:217–227CrossRefGoogle Scholar
  65. Scott GR, Sloman KA (2004) The effects of environmental pollutants on complex fish behaviour: integrating behavioural and physiological indicators of toxicity. Aquat Toxicol 68:369–392CrossRefGoogle Scholar
  66. Shepherd GM (1988) Neurobiology. Oxford University Press, New York, p 689Google Scholar
  67. Shimizu N, Ogino C, Kawanishi T, Hayashi Y (2002) Fractal analysis of Daphnia motion for acute toxicity bioassay. Environ Toxicol 17:441–448CrossRefGoogle Scholar
  68. Sørensen FF, Bayley M, Baatrup E (1995) The effects of sub-lethal dimethoate exposure on the locomotor behaviour of the collembolan Folsomia candida (Isotomidae). Environ Toxicol Chem 14:1587–1590CrossRefGoogle Scholar
  69. Steinberg CEW, Lorenz R, Spieser OH (1995) Effects of atrazine on swimming behaviour of zebrafish, Brachydanio rerio. Water Res 29:981–985CrossRefGoogle Scholar
  70. Tahedl H, Häder DP (1999) Fast examination of water quality using the automatic biotest ECOTOX based on the movement behaviour of a freshwater flagellate. Wat Res 33:426–432CrossRefGoogle Scholar
  71. Tahedl H, Häder DP (2001) Automated biomonitoring using real time movement analysis of Euglena gracilis. Ecotoxicol Environ Saf 48(2):161–169CrossRefGoogle Scholar
  72. Tay KL, Doe KG, Wade SJ, Vaughan DA, Berrigan RE, Moore MJ (1992) Sediment bioassessment in Halifax Harbour. Environ Toxicol Chem 11:1567–1581CrossRefGoogle Scholar
  73. Thomas KV, Fileman TW, Readman JW, Waldock MJ (2001) Antifouling paint booster biocides in the UK coastal environment and potential risks of biological effects. Mar Poll Bull 42(8):677–688CrossRefGoogle Scholar
  74. Thompson B, Bay S, Greenstein D, Laughlin J (1991) Sublethal effects of hydrogen-sulfide in sediments on the urchin Lytechinus-pictus. Mar Environ Res 31(4):309–321CrossRefGoogle Scholar
  75. Vogl C, Grillitsch B, Wytek R, Hunrich Spieser O, Scholz W (1999) Qualification of spontaneous undirected locomotor behavior of fish for sublethal toxicity testing. Part I. Variability of measurement parameters under general test conditions. Environ Toxicol Chem 18(12):2736–2742CrossRefGoogle Scholar
  76. Walker CH (1998) The use of biomarkers to measure the interactive effects of chemicals. Ecotoxicol Environ Saf 40:65–70CrossRefGoogle Scholar
  77. Wallace WG, Estephan A (2004) Differential susceptibility of horizontal and vertical swimming activity to cadmium exposure in a gammaridean amphipod (Gammarus lawrencianus). Aquat Toxicol 69:289–297CrossRefGoogle Scholar
  78. Ward LA, Montagna PA, Kalke RD, Buskey EJ (2000) Sublethal effects of Texas brown tide on Streblospio benedicti (Polychaeta) larvae. J Exp Mar Biol Ecol 248:121–129CrossRefGoogle Scholar
  79. Wedderburn J, McFadzen I, Sanger RC, Beesley A, Heath C, Hornsby M, Lowe D (2000) The field application of cellular and physiological biomarkers, in the mussel Mytilus edulis, in conjunction with early life stage bioassays and adult histopathology. Mar Poll Bull 40(3):257–267CrossRefGoogle Scholar
  80. Wolf G, Scheunders P, Selens M (1998) Evaluation of the swimming activity of Daphnia magna by image analysis after administration of sublethal cadmium concentrations. Comp Biochem Physiol A 120:99–105CrossRefGoogle Scholar
  81. Wu RSS, Lam PKS, Zhou BS (1997a) Effects of two oil dispersant on phototaxis and swimming behaviour of barnacle larvae. Hydrobiologia 352:9–16CrossRefGoogle Scholar
  82. Wu RSS, Lam PKS, Zhou BS (1997b) A settlement inhibition assay with cyprid larvae of the barnacle Balanus amphitrite. Chemosphere 35:1867–1874CrossRefGoogle Scholar
  83. Wu RSS, Lam PKS, Zhou BS (1997c) A phototaxis inhibition assay using barnacle larvae. Environ Toxic Water 12(3):231–236CrossRefGoogle Scholar
  84. Yachida M, Asada M, Tsuji S (1981) Automatic analysis of moving images. IEEE Trans Pat Anal Mach Intel PAMI 3:12–20CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • M. Faimali
    • 1
    Email author
  • F. Garaventa
    • 1
  • V. Piazza
    • 1
  • G. Greco
    • 1
  • C. Corrà
    • 1
  • F. Magillo
    • 1
  • M. Pittore
    • 2
  • E. Giacco
    • 3
  • L. Gallus
    • 3
  • C. Falugi
    • 3
  • G. Tagliafierro
    • 3
  1. 1.Institute of Marine ScienceNational Council of Researches (CNR)GenovaItaly
  2. 2.e-magine IT Computer Vision Image ProcessingGenovaItaly
  3. 3.Department of BiologyUniversity of GenoaGenovaItaly

Personalised recommendations