Skip to main content

Advertisement

SpringerLink
Log in

Encapsulation of live marine bacteria for use in aquaculture facilities and process evaluation using response surface methodology

  • Biotechnological products and process engineering
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

New strategies are being proposed in marine aquaculture to use marine bacteria as alternative to antibiotics, as nutritional additive or as immune-stimulant. These approaches are particularly promising for larval and juvenile cultures. In many cases, the bacteria are released in the seawater, where they have to be at appropriate concentrations. In addition, only low-cost technologies are sustainable for this industry, without any complex requirements for use or storage. In this work, we explore the possibilities of preservation of a potential marine probiotic bacterium (Phaeobacter PP-154) as a product suitable for use in marine aquaculture by addition to the seawater. A method which guaranteed the preservation of the viable marine bacteria in a saline medium and their rapid release in the seawater was searched for. In a previous step, classical procedures (freeze-drying and freezing) had been explored, but undesirable results of the interaction of the products obtained with natural seawater led to investigate alternatives. We report the results of the immobilization of the marine bacteria in calcium alginate beads. The final product complies the salinity which allows the requirements of the bacteria without interference with alginate in the formation of beads, and a balanced hardness to retain the bacteria and to be easily released in the marine aquaculture environment. The process was evaluated using the central composite rotatable design (CCRD), a standard response surface methodology (RSM).

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Becerra M, Baroli B, Fadda AM, Blanco Méndez J, González Siso MI (2001) Lactose bioconversion by calcium-alginate immobilization of Kluyveromyces lactis cells. Enzym Microb Technol 29:506–512

    Article  CAS  Google Scholar 

  • Box GEP, Hunter JS (1957) Multi-factor experimental designs for exploring response surfaces. Ann Math Statist 28:195–241

    Article  Google Scholar 

  • Brinkhoff T, Bach G, Heidorn T, Liang L, Schlingloff A, Simon M (2004) Antibiotic production by a Roseobacter clade-affiliated species from the German Wadden Sea and its antagonistic effects on indigenous isolates. Appl Environ Microbiol 70:2560–2565

    Article  CAS  Google Scholar 

  • Broeck G, Vandenheuvel D, Claes IJJ, Lebeer S, Kiekens F (2016) Drying techniques of probiotic bacteria as an important step towards the development of novel pharmabiotics. Int J Pharm 505:303–318

    Article  Google Scholar 

  • Bruhn JB, Gram L, Belas R (2007) Production of antibacterial compounds and biofilm formation by Roseobacter species are influenced by culture conditions. Appl Environ Microbiol 73:442–450

    Article  CAS  Google Scholar 

  • Callone E, Campostrini R, Carturan G, Cavazza A, Guzzon R (2008) Immobilization of yeast and bacteria cells in alginate microbeads coated with silica membranes: procedures, physico-chemical features and bioactivity. J Mater Chem 18:4839–4848

    Article  CAS  Google Scholar 

  • Chen YC (2003) Immobilized Isochrysis galbana (Haptophyta) for long-term storage and applications for feed and water quality control in clam (Meretrix lusoria) cultures. J Appl Phycol 15:439–444

    Article  Google Scholar 

  • Choi YS, Hong SR, Lee YM, Song KW, Park MH, Nam YS (1999) Study on gelatin-containing artificial skin: I. Preparation and characteristics of novel gelatin-alginate sponge. Biomaterials 20:409–417

    Article  CAS  Google Scholar 

  • Cordero H, Guardiola FA, Tapia-Paniagua ST, Cuesta A, Meseguer J, Balebona MC, Moriñigo MA, Esteban MA (2015) Modulation of immunity and gut microbiota after dietary administration of alginate encapsulated Shewanella putrefaciens Pdp11 to gilthead seabream (Sparus aurata L.). Fish Shellfish Immunol 45:608–618

    Article  CAS  Google Scholar 

  • de-Bashan LE, Bashan Y, Moreno M, Lebsky VK, Bustillos JJ (2002) Increased pigment and lipid content, lipid variety, and cell and population size of the microalgae Chlorella spp. when co-immobilized in alginate beads with the microalgae-growth-promoting bacterium Azospirillum brasilense. Can J Microbiol 48:514–521

    Article  CAS  Google Scholar 

  • Donati I, Holtan S, Mørch YA, Borgogna M, Dentini M, Skjåk-Bræk G (2005) New hypothesis on the role of alternating sequences in calcium-alginate gels. Biomacromolecules 6:1031–1040

    Article  CAS  Google Scholar 

  • Duarte JC, Rodrigues JAR, Moran PJS, Valença GP, Nunhez JR (2013) Effect of immobilized cells in calcium alginate beads in alcoholic fermentation. AMB Express 3:31

    Article  Google Scholar 

  • FAO (2011a) World Aquaculture 2010. FAO Fisheries and Aquaculture Department. Technical Paper. No. 500/1. Rome, Italy. 105 pp. (also available at http://www.fao.org/3/ba0132e/ba0132e.pdf)

  • FAO (2011b). Review of the state of world marine fishery resources. FAO Fisheries and Aquaculture Technical Paper No. 569. Rome, Italy. 2011. 334 pp. (also available at http://www.fao.org/3/i2389e/i2389e.pdf)

  • FAO (2016a). Aquaculture Big Numbers, by Michael Phillips, Rohana P. Subasinghe, Nhuong Tran, Laila Kassam and Chin Yee Chan. FAO Fisheries and Aquaculture Technical Paper No. 601. Rome, Italy. (also available at http://www.fao.org/3/a-i6317e.pdf)

  • FAO (2016b). FAO yearbook. Fishery and Aquaculture Statistics. 2014. Rome, Italy. (also available at http://www.fao.org/3/a-i5716t.pdf)

  • Feistel R, Wright DG, McDougall TJ (2008) The composition of standard seawater and the definition of the reference-composition salinity scale. Deep-Sea Res I 55:50–72

    Article  Google Scholar 

  • Gillet F, Roisin C, Fliniaux MA, Jacquin-Dubreuil A, Barbotin JN, Nava-Saucedo JE (2000) Immobilization of Nicotiana tabacum plant cell suspensions within calcium alginate gel beads for the production of enhanced amounts os scopolin. Enzym Microb Technol 26:229–234

    Article  Google Scholar 

  • Goh CH, Heng PWS, Chan LW (2012) Alginates as a useful natural polymer for microencapsulation and therapeutic applications. Carbohydr Polym 88:1–12

    Article  CAS  Google Scholar 

  • Hoseinifar SH, Sun YZ, Wang A, Zhou Z (2018) Probiotics as means of diseases control in aquaculture, a review of current knowledge and future perspectives. Front Microbiol 9:2429

    Article  Google Scholar 

  • Kakita H, Kamishima H (2008) Some properties of alginate gels derived from algal sodium alginate. J Appl Phycol 20:543–549

    Article  Google Scholar 

  • Kierstan M, Bucke C (1977) The immobilization of microbial cells, subcellular organelles, and enzymes in calcium alginate gels. Biotechnol Bioeng 19:387–397

    Article  CAS  Google Scholar 

  • Krasaekoopt W, Bhandari B, Deeth H (2003) Evaluation of encapsulation techniques of probiotics for yoghurt. Int Dairy J 13:3–13

    Article  CAS  Google Scholar 

  • Lutz C, Thomas T, Steinberg P, Kjelleberg S, Egan S (2016) Effect of interspecific competition on trait variation in Phaeobacter inhibens biofilms. Environ Microbiol 18:1635–1645

    Article  Google Scholar 

  • Mallón R, Barros P, Luzardo A, González ML (2007) Encapsulation of moss buds: an efficient method for the in vitro conservation and regeneration of the endangered moss Splanchnum ampullaceum. Plant Cell Tissue Organ Cult 88:41–49

    Article  Google Scholar 

  • Martin MJ, Lara-Villoslada F, Ruiz MA, Morales ME (2015) Microencapsulation of bacteria: a review of different technologies and their impact on the probiotic effects. Innov Food Sci Emerg Technol 27:15–25

    Article  CAS  Google Scholar 

  • Martinsen A, Skjåk-Bræk G, Smidsrød O (1989) Alginate as immobilization material: I. Correlation between chemical and physical properties of alginate gel beads. Biotechnol Bioeng 33:79–89

    Article  CAS  Google Scholar 

  • Millero FJ, Feistel R, Wright DG, McDougall TJ (2008) The composition of standard seawater and the definition of the reference-composition salinity scale. Deep Sea Res Part 1: Oceanogr Res Pap 55:50–72

    Article  Google Scholar 

  • Mizumoto S, Shoda M (2007) Medium optimization of antifungal lipopeptide, iturin A, production by Bacillus subtilis in solid-state fermentation by response surface methodology. Appl Microbiol Biotechnol 76:101–108

    Article  CAS  Google Scholar 

  • Nimrat S, Boonthai T, Vuthiphandchai V (2011) Effects of probiotic forms, compositions of and mode of probiotic administration on rearing of Pacific white shrimp (Litopenaeus vannamei) larvae and postlarvae. Anim Feed Sci Technol 169:244–258

    Article  CAS  Google Scholar 

  • Pales Espinosa E, Barillé L, Allam B (2007) Use of encapsulated live microalgae to investigate pre-ingestive selection in the oyster Crassostrea gigas. J Exp Mar Biol Ecol 343:118–126

    Article  Google Scholar 

  • Prado S (2006) Microbiota asociada a criaderos de moluscos: patogénesis y probiosis. Ph.D. Thesis. University of Santiago de Compostela (in Spanish)

  • Prado S, Montes J, Romalde JL, Barja JL (2009) Inhibitory activity of Phaeobacter strains against aquaculture pathogenic bacteria. Int Microbiol 12:107–114

    PubMed  Google Scholar 

  • Prado S, Romalde JL, Barja JL (2010) Review of probiotics for use in bivalve hatcheries. Vet Microbiol 145:187–197

    Article  Google Scholar 

  • Privman V, Domanskyi S, Luz RA, Guz N, Glasser ML, Katz E (2016) Diffusion of oligonucleotides from with iron-cross-linked, polyelectrolyte-modified alginate beads: a model system for drug release. Chemphyschem 17:976–984

    Article  CAS  Google Scholar 

  • Rathore S, Desai PM, Liew CV, Chan LW, Heng PWS (2013) Microencapsulation of microbial cells. J Food Eng 116:369–381

    Article  CAS  Google Scholar 

  • Romalde JL, Luzardo-Álvarez A, Ravelo C, Toranzo AE, Blanco-Méndez J (2004) Oral immunization using alginate microparticles as a useful strategy for booster vaccination against fish lactoccocosis. Aquaculture 236:119–129

    Article  CAS  Google Scholar 

  • Rosas-Ledesma P, León-Rubio JM, Alarcón FJ, Moriñigo MA, Balebona MC (2012) Calcium alginate capsules for oral administration of fish probiotic bacteria: assessment of optimal conditions for encapsulation. Aquac Res 43:106–116

    Article  Google Scholar 

  • Sachan NK, Pushkar S, Jha A, Bhattcharya A (2009) Sodium alginate: the wonder polymer for controlled drug delivery. J Pharm Res 2:1191–1199

    Google Scholar 

  • Tagliabue A, Bowie AR, Boyd PW, Buck KN, Johnson KS, Saito MA (2017) The integral role of iron in ocean biogeochemistry. Nature 543:51–59

    Article  CAS  Google Scholar 

  • Vemmer M, Patel AV (2013) Review of encapsulation methods suitable for microbial biological control agents. Biol Control 67:380–389

    Article  CAS  Google Scholar 

  • Westman JO, Ylitervo P, Franzén CJ, Taherzadeh MJ (2012) Effects of encapsulation of microorganisms on product formation during microbial fermentations. Appl Microbiol Biotechnol 96:1441–1454

    Article  CAS  Google Scholar 

  • Zhang Q, Chen T, Yang S, Wang X, Guo H (2013) Response surface methodology to design a selective enrichment broth for a rapid detection of Salmonella spp. by SYBR Green I real-time PCR. Appl Microbiol Biotechnol 97:4149–4158

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Rute Irgang for her technical help.

Funding

This work was supported by BAPOLABinHATCH (AGL2017-86183-R), Ministerio de Ciencia, Innovación y Universidades (Gobierno de España), co-funded with European Union ERDF funds. The support of Xunta de Galicia (Spain) with grant ED431C 2018/18 is also acknowledged.

Author information

Authors and Affiliations

  1. Dpto. Microbiología y Parasitología, CIBUS-Facultad de Biología, Universidad de Santiago de Compostela, 15782, Santiago de Compostela (A Coruña), Spain

    Susana Prado, Juan L. Barja & Javier Dubert

  2. Dpto. Farmacología, Farmacia y Tecnología Farmacéutica. Facultad de Ciencias, Universidad de Santiago de Compostela, 27002, Lugo, Spain

    Asteria Luzardo & José Blanco

Authors
  1. Susana Prado
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Juan L. Barja
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Asteria Luzardo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Javier Dubert
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. José Blanco
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Susana Prado.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(PDF 249 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Prado, S., Barja, J.L., Luzardo, A. et al. Encapsulation of live marine bacteria for use in aquaculture facilities and process evaluation using response surface methodology. Appl Microbiol Biotechnol 104, 1993–2006 (2020). https://doi.org/10.1007/s00253-019-10332-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-019-10332-0

Keywords

Search

Navigation