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Preparation and Characterization of Alginate Microparticles Containing a Model Protein for Oral Administration in Gnotobiotic European Sea Bass (Dicentrarchus labrax) Larvae

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Abstract

Aquaculture is the fastest growing animal production sector. However, the production of marine fish is still hampered by the high mortality rate in the first few weeks after hatching. Mortality in larvae is often caused by microbial infections. Today, the incorporation of immunostimulants into microparticles provides us new tools to enhance disease resistance in marine larviculture. In this study, we prepared alginate microparticles loaded with the model antigen fluorescein isothiocyanate conjugated-bovine serum albumin. Optimum concentrations of alginate and CaCl2, the correct alginate viscosity and the appropriate preparatory conditions led to the creation of desirable microparticles with the correct size for oral feeding in gnotobiotic European sea bass larvae. The prepared alginate microparticles were stable in sea water and were successfully ingested by gnotobiotic sea bass larvae at day after hatching 7 without causing any negative effects. Results suggest the suitability of this drug delivery system for targeting the innate immune system of fish larvae in order to enhance disease resistance and thus reduce mortality in larviculture.

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References

  • Chen SC, Wu YC, Mi FL, Lin YH, Yu LC, Sung HW (2004) A novel pH-sensitive hydrogel composed of N,O-carboxymethyl chitosan and alginate cross-linked by genipin for protein drug delivery. J Control Release 96:285–300

    Article  CAS  PubMed  Google Scholar 

  • Daemi H, Barikani M (2012) Synthesis and characterization of calcium alginate nanoparticles, sodium homopolymannuronate salt and its calcium nanoparticles. Sci Iran 19:2023–2028

    Article  CAS  Google Scholar 

  • Defoirdt T, Boon N, Bossier P, Verstraete W (2004) Disruption of bacterial quorum sensing: an unexplored strategy to fight infections in aquaculture. Aquaculture 240:69–88

    Article  Google Scholar 

  • Dierckens K, Rekecki A, Laureau S, Sorgeloos P, Boon N, Van den Broeck W, Bossier P (2009) Development of a bacterial challenge test for gnotobiotic sea bass (Dicentrarchus labrax) larvae. Environ Microbiol 11:526–533

    Article  CAS  PubMed  Google Scholar 

  • Draget KI (2009) Alginates. In: Phillips GO, Williams PA (eds) Handbook of Hydrocolloids, May 2009. Woodhead Publishing, Cambridge, pp 379–395

    Google Scholar 

  • Ertesvåg H, Valla S (1998) Biosynthesis and applications of alginates. Polym Degrad Stab 59:85–91

    Article  Google Scholar 

  • Fontagné S, Robin J, Corraze G, Bergot P (2000) Growth and survival of European sea bass (Dicentrarchus labrax) larvae fed from first feeding on compound diets containing medium-chain triacylglycerols. Aquaculture 190:261–271

    Article  Google Scholar 

  • Gacesa P (1988) Alginates. Carbohydr Polym 8:161–182

    Article  CAS  Google Scholar 

  • George M, Abraham TE (2006) Polyionic hydrocolloids for the intestinal delivery of protein drugs: alginate and chitosan—a review. J Control Release 114:1–14

    Article  CAS  PubMed  Google Scholar 

  • Gombotz WR, Wee SF (1998) Protein release from alginate matrices. Adv Drug Deliv Rev 31:267–285

    Article  CAS  PubMed  Google Scholar 

  • Hu B, Phuoc LH, Sorgeloos P, Bossier P (2014) Bacterial HSP70 (DnaK) is an efficient immune stimulator in Litopenaeus vannamei. Aquaculture 418–419:87–93

    Article  Google Scholar 

  • Jeffery H, Davis SS, O’Hagan DT (1993) The preparation and characterization of poly(lactide-co-glycolide) microparticles. II. The entrapment of a model protein using a (water-in-oil-in)-water emulsion solvent evaporation technique. Pharm Res 10:362–368

    Article  CAS  PubMed  Google Scholar 

  • Joosten PHM, Tiemersma AE, Threels A, Caumartin-Dhieux C, Rombout JHWM (1997) Oral vaccination of fish against Vibrio anguillarum using alginate microparticles. Fish Shellfish Immunol 7:471–485

    Article  Google Scholar 

  • Lemoine D, Wauters F, Bouchend’homme S, Préat V (1998) Preparation and characterization of alginate microspheres containing a model antigen. Int J Pharm 176:9–19

    Article  CAS  Google Scholar 

  • Liu L, Liu S, Ng SY, Froix M, Ohno T, Heller J (1997) Controlled release of interleukin-2 for tumour immunotherapy using alginate/chitosan porous microspheres. J Control Release 43:65–74

    Article  CAS  Google Scholar 

  • Marques A, François J-M, Dhont J, Bossier P, Sorgeloos P (2004) Influence of yeast quality on performance of gnotobiotically grown Artemia. J Exp Mar Bio Ecol 310:247–264

    Article  Google Scholar 

  • Moretti A, Fernandez-Criado MP, Cittolin G, Guidastri R (1999) Manual on hatchery production of seabass and gilthead seabream, vol 1. FAO, Rome, pp 1–194

  • Nikolopoulou D, Moutou KA, Fountoulaki E, Venou B, Adamidou S, Alexis MN (2011) Patterns of gastric evacuation, digesta characteristics and pH changes along the gastrointestinal tract of gilthead sea bream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.). Comp Biochem Physiol A Physiol 158:406–414

  • Polk AE, Amsden B, Scarratt DJ, Okhamafe AO, Goosen MFA (1994) Oral delivery in aquaculture: controlled release of proteins from chitosan-alginate microcapsules. Aquac Eng 13:311–323

    Article  Google Scholar 

  • Reis CP, Neufeld RJ, Ribeiro AJ, Veiga F (2006) Nanoencapsulation I. Methods for preparation of drug-loaded polymeric nanoparticles. Nanomed Nanotechnol Biol Med 2:8–21

  • Rivas-Aravena A, Sandino AM, Spencer E (2013) Nanoparticles and microparticles of polymers and polysaccharides to administer fish vaccines. Biol Res 46:407–419

    Article  PubMed  Google Scholar 

  • Rodrigues AP, Hirsch D, Figueiredo HCP, Logato PVR, Moraes ÂM (2006) Production and characterisation of alginate microparticles incorporating Aeromonas hydrophila designed for fish oral vaccination. Process Biochem 41:638–643

    Article  CAS  Google Scholar 

  • Romalde JL, Luzardo-Alvárez 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 

  • Saraei F, Mohamadpour Dounighi N, Zolfagharian N, Moradi Bidhendi H, Khaki S, Inanlou F (2013) Design and evaluate alginate nanoparticles as a protein delivery system. Arch Razi Inst 68:139–146

    CAS  Google Scholar 

  • Silva PF (2007) Gnotobiotic growth model system for Dicentrarchus labrax larvae. MSc. thesis, University of Algarve. Available from: http://hdl.handle.net/10400.1/625. Accessed 5 Nov 2015

  • Sosnik A (2014) Alginate particles as platform for drug delivery by the oral route: state-of-the-art. ISRN Pharm 2014:1–17

    Google Scholar 

  • Suksamran T, Opanasopit P, Rojanarata T, Ngawhirunpat T, Ruktanonchai U, Supaphol P (2009) Biodegradable alginate microparticles developed by electrohydrodynamic spraying techniques for oral delivery of protein. J Microencapsul 26:563–570

    Article  CAS  PubMed  Google Scholar 

  • Takka S, Acartürk F (1999) Calcium alginate microparticles for oral administration: I: effect of sodium alginate type on drug release and drug entrapment efficiency. J Microencapsul 16:275–290

    Article  CAS  PubMed  Google Scholar 

  • Tanaka H, Matsumura M, Veliky IA (1984) Diffusion characteristics of substrates in Ca-alginate gel beads. Biotechnol Bioeng 26:53–58

    Article  CAS  PubMed  Google Scholar 

  • Thu B, Gåserød O, Paus D, Mikkelsen A, Skjåk-bræk G, Toffanin R, Vittur F, Rizzo R (2000) Inhomogeneous alginate gel spheres: an assessment of the polymer gradients by induced x-ray emission, magnetic resonance microimaging, and mathematical modeling. Biopolymers 53:60–71

    Article  CAS  PubMed  Google Scholar 

  • Toranzo AE (2004) Report about fish bacterial diseases. In: Alvarez-Pellitero P, Barja JL, Basurco B, Berthe F, Toranzo AE (eds) Mediterranean Aquaculture Diagnostic Laboratories. CIHEAM, Zaragoza, pp 49–89

    Google Scholar 

  • Vadstein O, Bergh Ø, Gatesoupe F-J, Galindo-Villegas J, Mulero V, Picchietti S, Scapigliati G, Makridis P, Olsen Y, Dierckens K, Defoirdt T, Boon N, De Schryver P, Bossier P (2013) Microbiology and immunology of fish larvae. Rev Aquac 5:S1–S25

    Article  Google Scholar 

  • Walford J, Lim TM, Lam TJ (1990) Replacing live foods with microencapsulated diets in the rearing of seabass (Lates calcarifer) larvae: do the larvae ingest and digest protein-membrane microcapsules? Aquaculture 92:225–235

    Article  Google Scholar 

  • Yaacob EN, Vandenbosch D, Coenye T, Bajek A, Vanrompay D, Dierckens K, Bossier P (2016) Drinking activity and microparticle size selection in early post-hatching axenic European sea bass (Dicentrarchus labrax L.) larvae. Aquaculture 463:37–42

    Article  Google Scholar 

Download references

Acknowledgements

This research was supported and received financing from Ghent University (grant no. BOF-12-GOA-022). Eamy Nursaliza Yaacob received a doctoral scholarship from the Ministry of Higher Education Malaysia (MOHE) and the Universiti Malaysia Terengganu (UMT). Both UGent laboratories1,3 are members of the FWO Research Community: nutritional and microbial studies in larval aquaculture and this research was partially funded by the grant no. WO.030.15N. We would like to acknowledge Jorg De Smyter, Brigitte Van Moffaert and Tom Baelemans (Laboratory of Aquaculture & Artemia Reference Center; UGent), for their excellent technical support.

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Authors and Affiliations

  1. Laboratory of Aquaculture & Artemia Reference Center, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Eamy Nursaliza Yaacob, Kristof Dierckens & Peter Bossier

  2. School of Fisheries and Aquaculture Sciences, Universiti Malaysia Terengganu, 21030, Kuala Terengganu, Terengganu, Malaysia

    Eamy Nursaliza Yaacob

  3. Laboratory of Immunology and Animal Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Jens Goethals & Daisy Vanrompay

  4. Ecloserie Marine de Gravelines, Voie des Enrochements, 59820, Gravelines, France

    Aline Bajek

  5. Department of Pharmaceutics, Faculty of Pharmaceutical Science, Ghent University, Ottergemsesteenweg 460, 9000, Ghent, Belgium

    Bruno G. De Geest

Authors
  1. Eamy Nursaliza Yaacob
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  2. Jens Goethals
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  3. Aline Bajek
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  4. Kristof Dierckens
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  5. Peter Bossier
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  6. Bruno G. De Geest
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  7. Daisy Vanrompay
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Correspondence to Eamy Nursaliza Yaacob.

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Bruno G. De Geest and Daisy Vanrompay shared senior authorship

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Yaacob, E.N., Goethals, J., Bajek, A. et al. Preparation and Characterization of Alginate Microparticles Containing a Model Protein for Oral Administration in Gnotobiotic European Sea Bass (Dicentrarchus labrax) Larvae. Mar Biotechnol 19, 391–400 (2017). https://doi.org/10.1007/s10126-017-9758-4

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  • DOI: https://doi.org/10.1007/s10126-017-9758-4

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