, Volume 149, Issue 4, pp 604–619 | Cite as

Larval settlement of the common Australian sea urchin Heliocidaris erythrogramma in response to bacteria from the surface of coralline algae

  • Megan J. Huggett
  • Jane E. Williamson
  • Rocky de Nys
  • Staffan Kjelleberg
  • Peter D. Steinberg
Plant Animal Interactions


Bacterial biofilms are increasingly seen as important for the successful settlement of marine invertebrate larvae. Here we tested the effects of biofilms on settlement of the sea urchin Heliocidaris erythrogramma. Larvae settled on many surfaces including various algal species, rocks, sand and shells. Settlement was reduced by autoclaving rocks and algae, and by treatment of algae with antibiotics. These results, and molecular and culture-based analyses, suggested that the bacterial community on plants was important for settlement. To test this, approximately 250 strains of bacteria were isolated from coralline algae, and larvae were exposed to single-strain biofilms. Many induced rates of settlement comparable to coralline algae. The genus Pseudoalteromonas dominated these highly inductive strains, with representatives from Vibrio, Shewanella, Photobacterium and Pseudomonas also responsible for a high settlement response. The settlement response to different bacteria was species specific, as low inducers were also dominated by species in the genera Pseudoalteromonas and Vibrio. We also, for the first time, assessed settlement of larvae in response to characterised, monospecific biofilms in the field. Larvae metamorphosed in higher numbers on an inducing biofilm, Pseudoalteromonas luteoviolacea, than on either a low-inducing biofilm, Pseudoalteromonas rubra, or an unfilmed control. We conclude that the bacterial community on the surface of coralline algae is important as a settlement cue for H. erythrogramma larvae. This study is also an example of the emerging integration of molecular microbiology and more traditional marine eukaryote ecology.


  1. Abelson A, Denny M (1997) Settlement of marine organisms in flow. Annu Rev Ecol Syst 28:317–339CrossRefGoogle Scholar
  2. Altschul SF, Gish W, Miller W, Myers EE, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410PubMedGoogle Scholar
  3. Anderson MJ, Underwood AJ (1997) Effects of gastropod grazers on recruitment and succession of an estuarine assemblage: a multivariate and univariate approach. Oecologia 109:442–453CrossRefGoogle Scholar
  4. Barbieri E et al (2001) Phylogenetic characterization of epibiotic bacteria in the accessory nidamental gland and egg capsules of the squid Loligo peali (Cephalopoda: Loliginidae). Environ Microbiol 3:151–167PubMedCrossRefGoogle Scholar
  5. Boettcher AA, Targett NM (1998) Role of chemical inducers in larval metamorphosis of queen conch, Strombus gigas Linnaeus: relationship to other marine invertebrate systems. Biol Bull 194:132–142CrossRefGoogle Scholar
  6. Boxshall AJ (2000) The importance of flow and settlement cues to larvae of the abalone, Haliotis rufescens Swainson. J Exp Mar Biol Ecol 254:143–167PubMedCrossRefGoogle Scholar
  7. Browne KA, Zimmer RK (2001) Controlled release of a waterborne chemical signal stimulates planktonic larvae to settle. Biol Bull 200:87–91PubMedCrossRefGoogle Scholar
  8. Burke RD (1984) Pheromonal control of metamorphosis in the sand dollar, Dendraster excentricus. Science 225:442–443PubMedCrossRefGoogle Scholar
  9. Burke RD (1986) Pheromones and the gregarious settlement of marine invertebrate larvae. Bull Mar Sci 39:584–593Google Scholar
  10. Butman CA (1987) Larval settlement of soft-sediment invertebrates: the spatial scales of pattern explained by active habitat selection and the emerging role of hydrodynamical processess. Oceanogr Mar Biol Annu Rev 25:113–165Google Scholar
  11. Caley MJ, Carr ME, Hixon MA, Hughes TP, Jones GP, Menge BA (1996) Recruitment and the local dynamics of open marine populations. Annu Rev Ecol Syst 27:477–500CrossRefGoogle Scholar
  12. Callow ME et al (2002) Microtopographic cues for settlement of zoospores of the green fouling alga Enteromorpha. Biofouling 18:237–245CrossRefGoogle Scholar
  13. Clarke KR, Warwick RM (1994) Change in marine communities: an approach to statistical analysis and interpretation. Plymouth Marine Laboratory, PlymouthGoogle Scholar
  14. Dahllöf I, Baillie H, Kjelleberg S (2000) rpoB-based microbial community analysis avoids limitations inherent in 16S rRNA gene intraspecies heterogeneity. Appl Environ Microbiol 66:3376–3380PubMedCrossRefGoogle Scholar
  15. Dahms H-U, Dobretsov S, Qian P-Y (2004) The effect of bacterial and diatom biofilms on the settlement of the bryozoan Bugula neritina. J Exp Mar Biol Ecol 313:191–209CrossRefGoogle Scholar
  16. Daume S, Brand-Gardner S, Woelkerling WJ (1999) Settlement of abalone larvae (Haliotis laevigata Donovan) in response to non-geniculate coralline red algae (Corallinales, Rhodophyta). J Exp Mar Biol Ecol 234:125–143CrossRefGoogle Scholar
  17. Daume S, Krsinich A, Farrell S, Gervis M (2000) Settlement, early growth and survival of Haliotis rubra in response to different algal species. J Appl Phycol 12:479–488CrossRefGoogle Scholar
  18. Fusetani N (1997) Marine natural products influencing larval settlement and metamorphosis of benthic invertebrates. Curr Org Chem 1:127–115Google Scholar
  19. Gallardo WG, Buen SA (2003) Evaluation of mucus, Navicula, and mixed diatoms as larval settlement inducers for the tropical abalone Haliotis asinina. Aquaculture 221:357–364CrossRefGoogle Scholar
  20. Gordon N, Shpigel M, Harpaz S, Lee JJ, Neori A (2004) The settlement of abalone (Haliotis discus hannai) larvae on culture layers of different diatoms. J Shellfish Res 23:561–568Google Scholar
  21. Gosselin P, Jangoux M (1996) Induction of metamorphosis in Paracentrotus lividus larvae (Echinodermata, Echinoidea). Oceanol Acta 19:293–296Google Scholar
  22. Hadfield MG, Paul VJ (2001) Natural chemical cues for settlement and metamorphosis of marine invertebrate larvae. In: McClintock JB, Baker JB (eds) Marine chemical ecology. CRC, Boca Raton, Fla., pp 431–461Google Scholar
  23. Harder T, Lam C, Qian PY (2002) Induction of larval settlement in the polychaete Hydroides elegans by marine biofilms: an investigation of monospecific diatom films as settlement cues. Mar Ecol Prog Ser 229:105–112CrossRefGoogle Scholar
  24. Hentschel U, Schmid M, Wagner M, Fieseler L, Gernert C, Hacker J (2001) Isolation and phylogenetic analysis of bacteria with antimicrobial activities from the Mediterranean sponges Aplysina aerophoba and Aplysina cavernicola. FEMS Microbiol Ecol 35:305–312PubMedCrossRefGoogle Scholar
  25. Hofmann DK, Brand U (1987) Induction of metamorphosis in the symbiotic Scyphozoan Cassiopea andromeda: role of marine bacteria and biochemicals. Symbiosis 4:99–116Google Scholar
  26. Holmström C, Kjelleberg S (1999) Marine Pseudoalteromonas species are associated with higher organisms and produce biologically active extracellular agents. FEMS Microbiol Ecol 30:285–293PubMedGoogle Scholar
  27. Holmström C, Kjelleberg S (2000) Bacterial interactions with marine fouling organisms. In: Evans LV (eds) Biofilms: recent advances in their study and control. Harwood Academic Publishers, Australia, pp 101–115Google Scholar
  28. Holmström C, Rittschof D, Kjelleberg S (1992) Inhibition of settlement by larvae of Balanus amphitrite and Ciona intestinalis by a surface-colonizing marine bacterium. Appl Environ Microbiol 58:2111–2115PubMedGoogle Scholar
  29. Huang S, Hadfield MG (2003) Composition and density of bacterial biofilms determine larval settlement of the polycheate Hydroides elegans. Mar Ecol Prog Ser:161–172Google Scholar
  30. Huggett MJ, de Nys R, Williamson JE, Heasman M, Steinberg PD (2005) Settlement of larval blacklip abalone, Haliotis rubra, in response to red and green macroalgae. Mar Biol 147:1155–1163CrossRefGoogle Scholar
  31. Johnson CR, Sutton DC (1994) Bacteria on the surface of crustose coralline algae induce metamorphosis of the crown-of-thorns starfish Acanthaster planci. Mar Biol 120:305–310CrossRefGoogle Scholar
  32. Johnson CR, Muir DG, Reysenbach AL (1991a) Characteristic bacteria associated with surfaces of coralline algae: a hypothesis for bacterial induction of marine invertebrate larvae. Mar Ecol Prog Ser 74:281–294CrossRefGoogle Scholar
  33. Johnson CR, Sutton DC, Olson RR, Giddins R (1991b) Settlement of crown-of-thorns starfish: role of bacteria on surfaces of coralline algae and hypothesis of deep water recruitment. Mar Ecol Prog Ser 71:143–162CrossRefGoogle Scholar
  34. Keesing JK (2001) The ecology of Heliocidaris erythrogramma. In: Lawrence JM (eds) Edible sea urchins: biology and ecology. Elsevier, New York, pp 261–270CrossRefGoogle Scholar
  35. Keough MJ, Raimondi PT (1995) Responses of settling invertebrate larvae to bioorganic films: effects of different types of films. J Exp Mar Biol Ecol 185:235–253CrossRefGoogle Scholar
  36. Kobak J (2001) Light, gravity and conspecifics as cues to site selection and attachment behaviour of juvenile and adult Dreissena polymorpha Pallas, 1771. J Mollusc Stud 67:183–189CrossRefGoogle Scholar
  37. Krug PJ (2001) Bet-hedging dispersal strategy of a specialist marine herbivore: settlement dimorphism among sibling larvae of Alderia modesta. Mar Ecol Prog Ser 213:177–192CrossRefGoogle Scholar
  38. Krug PJ, Manzi AE (1999) Waterborne and surface-associated carbohydrates as settlement cues for larvae of the specialist marine herbivore Alderia modesta. Biol Bull 197:94–103CrossRefGoogle Scholar
  39. Lamare MD, Barker MF (2001) Settlement and recruitment of the New Zealand sea urchin Evechinus chloroticus. Mar Ecol Prog Ser 218:153–166CrossRefGoogle Scholar
  40. Lau SK, Qian PY (2001) Larval settlment in the serpulid polycheate Hydroides elegans in response to bacterial films: an investigation of the nature of putitative larval settlement cue. Mar Biol 138:321–328CrossRefGoogle Scholar
  41. Lau SCK, Mak KKW, Chen F, Qian PY (2002) Bioactivity of bacterial strains isolated from marine biofilms in Hong Kong waters for the induction of larval settlement in the marine polychaete Hydroides elegans. Mar Ecol Prog Ser 226:301–310CrossRefGoogle Scholar
  42. Lau SCK, Harder T, Qian P (2003) Induction of larval settlement in the serpulid polycheate Hydroides elegans (Haswell): role of bacterial extracellular polymers. Biofouling 19:197–204PubMedCrossRefGoogle Scholar
  43. Lau SCK, Thiyagarajan V, Cheung SCK, Qian P-Y (2005) Roles of bacterial community composition in biofilms as a mediator for larval settlement of three marine invertebrates. Aquat Microb Ecol 38:41–51CrossRefGoogle Scholar
  44. Leitz T (1997) Induction and metamorphosis of cnidarian larvae: signals and signal transduction. Invertebr Reprod Dev 31:109–122Google Scholar
  45. Morse AC (1992) Role of algae in the recruitment of marine invertebrate larvae. In: John DM, Hawkins SJ, Price JH (eds) Plant–animal interactions in the marine benthos, vol 46. Clarendon Press, Oxford, pp 385–403Google Scholar
  46. Morse ANC, Froyd CA, Morse DE (1984) Molecules from cyanobacteria and red algae that induce settlement and metamorphosis in the mollusc Haliotis rufescens. Mar Biol 81:293–298CrossRefGoogle Scholar
  47. Neal AL, Yule AB (1994) The interaction between Elminius modestus Darwin cyprids and biofilms of Deleya marina Ncmb1877. J Exp Mar Biol Ecol 176:127–139CrossRefGoogle Scholar
  48. Negri AP, Webster NS, Hill RT, Heyward AJ (2001) Metamorphosis of broadcast spawning corals in response to bacteria isolated from crustose algae. Mar Ecol Prog Ser 223:121–131CrossRefGoogle Scholar
  49. Patil JS, Anil AC (2005) Influence of diatom exoploymers and biofilms on metamorphosis in the barnacle Balanus amphitrite. Mar Ecol Prog Ser 301:231–245CrossRefGoogle Scholar
  50. Pawlik JR (1992) Chemical ecology of the settlement of marine invertebrates. Oceanogr Mar Biol Annu Rev 30:273–335Google Scholar
  51. Pearce CM, Scheibling RE (1991) Effect of macroalgae, microbial films, and conspecifics on the induction of metamorphosis of the green sea urchin Strongylocentrotus droebachiensis (Muller). J Exp Mar Biol Ecol 147:147–162CrossRefGoogle Scholar
  52. Pechenik JA (1999) On the advantages and disadvantages of larval stages in benthic marine invertebrate life cycles. Mar Ecol Prog Ser 177:269–297CrossRefGoogle Scholar
  53. Prescott LM, Harley JP, Klein DA (1995) Microbiology, 3rd edn. Brown, DubuqueGoogle Scholar
  54. Raimondi PT, Morse ANC (2000) The consequences of complex larval behaviour in a coral. Ecology 81(11):3193–3211CrossRefGoogle Scholar
  55. Roberts R (2001) A review of settlement cues for larval abalone (Haliotis spp.). J Shellfish Res 20:571–586Google Scholar
  56. Rodriguez SR, Ojeda FP, Inestrosa NC (1993) Settlement of benthic marine invertebrates. Mar Ecol Prog Ser 97:193–207CrossRefGoogle Scholar
  57. Rodriguez SR, Riquelme C, Campos EO, Chavez P, Brandan E, Inestrosa NC (1995) Behavioral responses of Concholepas concholepas (Bruguiere, 1789) larvae to natural and artificial settlement cues and microbial films. Biol Bull 189:272–279CrossRefGoogle Scholar
  58. Steinberg PD, de Nys R (2002) Chemical mediation of colonization of seaweed surfaces. J Phycol 38:621–629CrossRefGoogle Scholar
  59. Steinberg PD, de Nys R, Kjelleberg S (2001) Chemical mediation of surface colonization. In: McClintock JB, Baker BJ (eds) Marine chemical ecology. CRC, Boca Raton, Fla., pp 355–387Google Scholar
  60. Swanson RL, Williamson JE, de Nys R, Kumar N, Bucknall MP, Steinberg PD (2004) Induction of settlement of larvae of the sea urchin Holopneustes purpurascens by histamine from a host alga. Biol Bull 206:161–172PubMedCrossRefGoogle Scholar
  61. Switzer-Dunlap M (1978) Larval biology and metamorphosis of Aplysiid gastropods. In: Chia F-S, Rice ME (eds) Settlement and metamorphosis of marine invertebrate larvae. Elsevier, New York, pp 197–206Google Scholar
  62. Switzer-Dunlap M, Hadfield M (1977) Observations on development, larval growth and metamorphosis of four species of Aplysiidae (Gastropa: Opisthobranchia) in laboratory culture. J Exp Mar Biol Ecol 29:245–261CrossRefGoogle Scholar
  63. Szewzyk U, Holstrom C, Wrangstadh M, Samuelsson M-O, Maki JS, Kjelleberg S (1991) Relevance of the exopolysaccharide of marine Pseudomonas sp. strain S9 for the attachment of Ciona intestinalis larvae. Mar Ecol Prog Ser 75:259–265CrossRefGoogle Scholar
  64. Thompson RC, Roberts MF, Norton TA, Hawkins SJ (2000) Feast or famine for intertidal grazing molluscs: a mis-match between seasonal variations in grazing intensity and the abundance of microbial resources. Hydrobiologia 440:357–367CrossRefGoogle Scholar
  65. Thorson G (1950) Reproductive and larval ecology of marine bottom invertebrates. Biol Rev 25:1–45CrossRefGoogle Scholar
  66. Towbridge CD (1992) Phenology and demography of a marine specialist herbivore: Placida dendritica (Gastropoda: Opisthobrancha) on the central coast of Oregon. Mar Biol 114:443–452CrossRefGoogle Scholar
  67. Unabia CRC, Hadfield MG (1999) Role of bacteria in larval settlement and metamorphosis of the polychaete Hydroides elegans. Mar Biol 133:55–64CrossRefGoogle Scholar
  68. Underwood AJ, Keough MJ (2000) Supply-side ecology: the nature and consequences of variations in recruitment of intertidal organisms. In: Bertness MD, Gaines SD, Hay ME (eds) Marine community ecology. Sinauer, Sunderland, Mass., pp 183–200Google Scholar
  69. Webster NS, Smith LD, Heyward AJ, Watts JEM, Webb RI, Blackall LL, Negri AJ (2004) Metamorphosis of a scleractinian coral in response to microbial biofilms. Appl Environ Microbiol 70:1213–1221PubMedCrossRefGoogle Scholar
  70. Weiner RM, Williams N, Birch G, Ramachandran C, Collwell RR (1989) Effect of biofilms of the marine bacterium Alteromonas colwelliana (LST) on set of oysters Crassostrea gigas Thunberg 1793 and Crassostrea virginica Gmelin 1791. J Shellfish Res 8:117–124Google Scholar
  71. Wieczorek SK, Todd CD (1998) Inhibition and facilitation of settlement of epifaunal marine invertebrate larvae by microbial biofilm cues. Biofouling 12:81–118CrossRefGoogle Scholar
  72. Wilkinson L (1997) SYSTAT 7.0. SPSS, Chicago, Ill.Google Scholar
  73. Williams DHC, Anderson DT (1975) The reproductive system, embryonic development, larval development and metamorphosis of the sea urchin Heliocidaris erythrogramma (Val.) (Echinoidea: Echinometridae). Aust J Zool 23:371–403CrossRefGoogle Scholar
  74. Williamson JE, de Nys R, Kumar N, Carson DG, Steinberg PD (2000) Induction of metamorphosis in the sea urchin Holopneustes purpurascens by a metabolite complex from the algal host Delisea pulchra. Biol Bull 198:332–345PubMedCrossRefGoogle Scholar
  75. Xue-wu L, Gordon ME (1987) Tissue and cell culture of New Zealand Pterocladia and Porphyra species. Hydrobiologia 151/152:147–154CrossRefGoogle Scholar
  76. Zhao B, Qian PY (2002) Larval settlement and metamorphosis in the slipper limpet Crepidula onyx (Sowerby) in response to conspecific cues and the cues from biofilm. J Exp Mar Biol Ecol 269:39–51CrossRefGoogle Scholar
  77. Zobell CE, Allen EC (1935) The significance of marine bacteria in the fouling of submerged surfaces. J Bacteriol 29:230–251Google Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Megan J. Huggett
    • 1
    • 2
    • 5
  • Jane E. Williamson
    • 1
    • 3
  • Rocky de Nys
    • 1
    • 4
  • Staffan Kjelleberg
    • 1
  • Peter D. Steinberg
    • 1
    • 2
  1. 1.Centre for Marine Biofouling and BioInnovationUniversity of New South WalesSydneyAustralia
  2. 2.School of Biological, Earth and Environmental SciencesUniversity of New South WalesSydneyAustralia
  3. 3.Department of Biological ScienceMacquarie UniversitySydneyAustralia
  4. 4.School of Marine Biology and AquacultureJames Cook UniversityTownsvilleAustralia
  5. 5.Kewalo Marine LaboratoryUniversity of HawaiiHonoluluUSA

Personalised recommendations