Skip to main content
SpringerLink
Log in

Pathogenicity of a Highly Exopolysaccharide-producing Halomonas Strain Causing Epizootics in Larval Cultures of the Chilean Scallop Argopecten purpuratus (Lamarck, 1819)

  • Original Article
  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

Mass mortalities of larval cultures of Chilean scallop Argopecten purpuratus have repeatedly occurred in northern Chile, characterized by larval agglutination and accumulation in the bottom of rearing tanks. The exopolysaccharide slime (EPS) producing CAM2 strain was isolated as the primary organism from moribund larvae in a pathogenic outbreak occurring in a commercial hatchery producing larvae of the Chilean scallop Argopecten purpuratus located in Bahía Inglesa, Chile. The CAM2 strain was characterized biochemically and was identified by polymerase chain reaction amplification of 16S rRNA as Halomonas sp. (Accession number DQ885389.1). Healthy 7-day-old scallop larvae cultures were experimentally infected for a 48-h period with an overnight culture of the CAM2 strain at a final concentration of ca. 105 cells per milliliter, and the mortality and vital condition of larvae were determined by optical and scanning electron microscopy (SEM) to describe the chronology of the disease. Pathogenic action of the CAM2 strain was clearly evidenced by SEM analysis, showing a high ability to adhere and detach larvae velum cells by using its “slimy” EPS, producing agglutination, loss of motility, and a posterior sinking of scallop larvae. After 48 h, a dense bacterial slime on the shell surface was observed, producing high percentages of larval agglutination (63.28 ± 7.87%) and mortality (45.03 ± 4.32%) that were significantly (P < 0.05) higher than those of the unchallenged control cultures, which exhibited only 3.20 ± 1.40% dead larvae and no larval agglutination. Furthermore, the CAM2 strain exhibited a high ability to adhere to fiberglass pieces of tanks used for scallop larvae rearing (1.64 × 105 cells adhered per square millimeters at 24 h postinoculation), making it very difficult to eradicate it from the culture systems. This is the first report of a pathogenic activity on scallop larvae of Halomonas species, and it prompts the necessity of an appraisal on biofilm-producing bacteria in Chilean scallop hatcheries.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Anderson C, Atlar M, Callow M, Candries M, Townsin RL (2003) The development of foul-release coatings for seagoing vessels. J Mar Des Operat B4:11–23

    Google Scholar 

  2. Amaro AM, Fuentes MS, Ogalde SR, Venegas JA, Suárez-Isla BA (2005) Identification and characterization of potentially algal-lytic marine bacteria strongly associated with the toxic dinoflagellate Alexandrium catenella. J Eukaryot Microbiol 52:191–200

    PubMed  Google Scholar 

  3. Arahal DR, Ventosa A (2006) The Family Halomonadaceae. In: Dworkin M, Falkow S, Rosenberg E, Schleifer K-H, Stackebrandt E (eds) The prokaryotes. A handbook on the biology of bacteria, vol. 6. Proteobacteria: gamma subclass. 3rd edn. Springer, New York, pp 811–835

    Google Scholar 

  4. Arahal DR, Ludwig W, Schleifer K-H, Ventosa A (2002) Phylogeny of the family Halomonadaceae based on 23S and 16S rDNA sequence analyses. Int J Syst Evol Microbiol 52:241–249

    PubMed  CAS  Google Scholar 

  5. Armstrong E, Boyd KG, Burgess JG (2000) Prevention of marine biofouling using natural compounds from marine organisms. Biotechnol Annu Rev 6:221–241

    Article  PubMed  CAS  Google Scholar 

  6. Avendaño RE, Riquelme CE, Escribano R, Reyes N (2001) Sobrevivencia y crecimiento de post-larvas de Argopecten purpuratus (Lamarck, 1819) em Bahia Inglesa, Chile: efectos del origen, distribución en la bahía y bacterioflora larval. Rev Chil Hist Nat 74:669–679

    Article  Google Scholar 

  7. Barrow GI, Feltham RKA (1993) Cowan and Steel’s manual for the identification of medical bacteria, 3rd edn. Cambridge University Press, Cambridge 331 p

    Google Scholar 

  8. Bassler BL (1999) How bacteria talk to each other: regulation of gene expression by quorum sensing. Curr Opin Microbiol 2:582–587

    Article  PubMed  CAS  Google Scholar 

  9. Bouchotroch S, Quesada E, Izquierdo I, Rodríguez M, Béjar V (2000) Bacterial exopolysaccharides produced by newly discovered bacteria belonging to the genus Halomonas, isolated from hypersaline habitats in Morocco. J Ind Microbiol Biotech 24:374–378

    Article  CAS  Google Scholar 

  10. Clarkson N (2000) The antifouling potential of silicone elastomer polymers. In: Fingermann M, Nagabhushanam R, Thompson MF (eds) Recent advances in marine biotechnology, vol 3. Biofilms, bioadhesion, corrosion and biofouling. Science Publishers, USA, pp 147–171

    Google Scholar 

  11. de Nys R, Steinberg PD (2002) Linking marine biology and biotechnology. Curr Opin Biotechnol 13:244–248

    Article  PubMed  CAS  Google Scholar 

  12. Dobretsov S, Dahms HU, YiLi H, Wahl M, Qian PY (2007) The effect of quorum sensing blockers on the formation of marine microbial communities and larval attachment. FEMS Microbiol Ecol 60:177–188

    Article  PubMed  CAS  Google Scholar 

  13. Elston RA (1984) Prevention and management of infectious diseases in intensive mollusc husbandry. J World Maric Soc 15:284–300

    Google Scholar 

  14. Farias A, Uriarte I, Castilla JC (1998) A biochemical study of the larval and postlarval stages of the Chilean scallop Argopecten purpuratus. Aquaculture 166:37–47

    Article  CAS  Google Scholar 

  15. Gherna LR (1994) Culture preservation. In: Gerhardt P, Murray REG, Wood WA, Krieg NR (eds) Methods for general and molecular bacteriology. American Society for Microbiology, Washington, DC, pp 278–292

    Google Scholar 

  16. Gómez-León J, Villamil L, Lemos ML, Novoa B, Figueras A (2005) Isolation of Vibrio alginolyticus and Vibrio splendidus from Aquacultured Carpet Shell Clam (Ruditapes decussatus) Larvae Associated with Mass Mortalities. Appl Environ Microbiol 71:98–104

    Article  PubMed  CAS  Google Scholar 

  17. Hansen GH, Sörheim R (1991) Improved method for phenotypical characterization of marine bacteria. J Microbiol Methods 13:231–241

    Article  Google Scholar 

  18. Hentzer M, Wu H, Andersen JB, Riedel K, Rasmussen TB, Bagge N, Kumar N, Schembri MA, Song Z, Kristoffersen P, Manefield M, Costerton JW, Molin S, Eberl L, Steinberg P, Kjelleberg S, Hoiby N, Givskov M (2003) Attenuation of Pseudomonas aeruginosa virulence by quorum sensing inhibitors. EMBO J 22:3803–3815

    Article  PubMed  CAS  Google Scholar 

  19. Jeffries V (1982) Three Vibrio strains pathogenic to larvae of Crassostrea virginica and Ostrea edulis. Aquaculture 29:201–206

    Article  Google Scholar 

  20. Kjelleberg S, Steinberg P, Givskov M, Gram L, Manefield M, de Nys R (1997) Do marine natural products interfere with prokaryotic AHL regulatory systems. Aquat Microb Ecol 13:85–93

    Article  Google Scholar 

  21. Lee OO, Qian P (2003) Chemical control of bacterial epibiosis and larval settlement of Hydroides elegans in red sponge Myscale adherens. Biofouling 19:171–180

    Article  PubMed  CAS  Google Scholar 

  22. Lodeiros C, Bolinches J, Dopazo C, Toranzo A (1987) Bacillary necrosis in hatcheries of Ostrea edulis in Spain. Aquaculture 65:15–29

    Article  Google Scholar 

  23. Mah T-FC, O’Toole GA (2001) Mechanisms of biofilm resistance to antimicrobial agents. Trends Microbiol 9:34–39

    Article  PubMed  CAS  Google Scholar 

  24. Maki JS, Ding L, Stokes J, Kavouras JH, Rittschof D (2000) Substratum/bacterial interactions and larval attachment: films and exopolysaccharides of Halomonas marina (ATCC 25374) and their effect on barnacle cyprid larvae, Balanus amphitrite Darwin. Biofouling 16:159–170

    CAS  Google Scholar 

  25. Mata JA, Martínez-Canovas J, Quesada E, Béjar V (2002) A detailed phenotypic characterisation of the type strains of Halomonas species. Syst Appl Microbiol 25:360–375

    Article  PubMed  CAS  Google Scholar 

  26. Miranda CD, Rojas R (2006) Copper accumulation by bacteria and transfer to scallop larvae. Mar Pollut Bull 52:293–300

    Article  PubMed  CAS  Google Scholar 

  27. Nevell TG, Edwards DP, Davies AJ, Pullin RA (1996) The surface properties of silicone elastomers exposed to seawater. Biofouling 10:199–212

    CAS  Google Scholar 

  28. Nicolas JL, Corre S, Gauthier G, Robert R, Ansquer D (1996) Bacterial problems associated with scallop Pecten maximus larval culture. Dis Aquat Org 27:67–76

    Article  Google Scholar 

  29. Norusis MJ (2004) Statistical Package for the Social Sciences (SPSS) 12.0 Statistical Procedures Companion. Prentice Hall, Englewood Cliffs, USA 601 p

    Google Scholar 

  30. Nottage A, Birkbeck TH (1986) Toxicity of marine bivalves of culture supernatant fluids of the bivalve-pathogenic Vibrio strain NCMB 1338 and other vibrios. J Fish Dis 9:249–256

    Article  Google Scholar 

  31. Nottage A, Birkbeck TH (1987) The role of toxins in Vibrio infections of bivalve molluscs. Aquaculture 67:244–246

    Article  Google Scholar 

  32. Olson ME, Ceri H, Morck DW, Buret AG, Read RR (2002) Biofilm bacteria: formation and comparative susceptibility to antibiotics. Can J Vet Res 66:86–92

    PubMed  Google Scholar 

  33. Prado S, Romalde JL, Montes J, Barja JL (2005) Pathogenic bacteria isolated from disease outbreaks in shellfish hatcheries. First description of Vibrio neptunius as an oyster pathogen. Dis Aquat Org 67:209–215

    Article  PubMed  CAS  Google Scholar 

  34. Prieur D, Nicolas JL, Plusquellec A, Vigneulle M (1990) Interaction between bivalve molluscs and bacteria in the marine environment. Oceanogr Mar Biol Annu Rev 28:277–352

    Google Scholar 

  35. Riquelme C, Chavez P, Morales Y, Hayashida G (1994) Evidence for parental bacterial transfer to larvae in Argopecten purpuratus (Lamarck, 1819). Biol Res 27:129–134

    Google Scholar 

  36. Riquelme C, Hayashida G, Toranzo A, Vilches J, Chavez P (1995) Pathogenicity studies on a Vibrio anguillarum-related (VAR) strain causing an epizootic in Argopecten purpuratus larvae cultured in Chile. Dis Aquat Org 22:135–141

    Article  Google Scholar 

  37. Riquelme C, Hayashida G, Vergara N, Vasquez A, Morales Y, Chavez P (1995) Bacteriology of the scallop Argopecten purpuratus (Lamarck, 1819) cultured in Chile. Aquaculture 138:49–60

    Article  Google Scholar 

  38. Riquelme C, Toranzo A, Barja J, Vergara N, Araya R (1996) Association of Aeromonas hydrophila and Vibrio alginolyticus with larval mortalities of scallop (Argopecten purpuratus). J Invert Pathol 67:213–218

    Article  Google Scholar 

  39. Robert R, Miner P, Nicolas JL (1996) Mortality control of scallop larvae in the hatchery. Aquac Int 4:305–313

    Article  Google Scholar 

  40. Rosenblum BB, Lee LG, Spurgeon SL, Khan SH, Menchen SM, Heiner CR, Chen SM (1997) New dye-labeled terminators for improved DNA sequencing patterns. Nucleic Acids Res 25:4500–4504

    Article  PubMed  CAS  Google Scholar 

  41. Sainz J, Maeda-Martínez A, Ascencio F (1998) Experimental vibriosis induction with Vibrio alginolyticus of larval of the Caterina scallop (Argopecten ventricosis=circularis) (Sowerby II, 1842). Microb Ecol 35:188–192

    Article  PubMed  Google Scholar 

  42. Schulze AD, Alabib AO, Tattersall-Sheldrakeb AR, Miller KM (2006) Bacterial diversity in a marine hatchery: balance between pathogenic and potentially probiotic bacterial strains. Aquaculture 256:50–73

    Article  Google Scholar 

  43. Valsecchi E (1998) Tissue boiling: a short-cut in DNA extraction for large-scale population screenings. Mol Ecol 7:1243–1245

    Article  PubMed  CAS  Google Scholar 

  44. Walne PR (1965) Observations on the influence of food supply and temperature on the feeding and growth of the larvae of Ostrea edulis L. Fish Invest II 24:1–45

    Google Scholar 

  45. Zar JH (1999) Biostatistical analysis, 4th edn. Prentice-Hall, Inc., Upper Saddle River, NJ, USA 931 p

    Google Scholar 

Download references

Acknowledgments

This study was partially supported by the Project Grant PT-10 from the Developing and Innovation Funding (FDI-CORFO) of Chile. We would like to acknowledge the cooperation of Chilean scallop hatchery managers Alejandro Abarca and Christian Tapia who, over many years, have freely collaborated with us to solve their problems. We also thank MSc. Marisol Romero from Electronic Microscopy Unit of the Universidad Católica del Norte for her expert technical assistance. The comments and suggestions of the reviewers are greatly appreciated as they helped to improve the presentation of this work.

Author information

Authors and Affiliations

  1. Laboratorio de Patobiología Acuática, Departamento de Acuicultura, Facultad de Ciencias del Mar, Universidad Católica del Norte, Coquimbo, Chile

    Rodrigo Rojas & Claudio D. Miranda

  2. Centro de Estudios Avanzados en Zonas Áridas (CEAZA), Coquimbo, Chile

    Claudio D. Miranda

  3. Laboratorio de Toxinas Marinas, Programa de Fisiología y Biofísica. Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile

    Ana María Amaro

Authors
  1. Rodrigo Rojas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Claudio D. Miranda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ana María Amaro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Claudio D. Miranda.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rojas, R., Miranda, C.D. & Amaro, A.M. Pathogenicity of a Highly Exopolysaccharide-producing Halomonas Strain Causing Epizootics in Larval Cultures of the Chilean Scallop Argopecten purpuratus (Lamarck, 1819). Microb Ecol 57, 129–139 (2009). https://doi.org/10.1007/s00248-008-9401-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-008-9401-z

Keywords

Search

Navigation