Skip to main content
SpringerLink
Log in

Responses of Atlantic cod Gadus morhua head kidney leukocytes to phytase produced by gastrointestinal-derived bacteria

  • Published:
Fish Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

This study identified phytase-producing bacteria that were previously isolated from the gastrointestinal tract of Atlantic cod, Gadus morhua and determined its effect on head kidney leukocytes. Out of the 216 bacterial strains tested, the two phytase producers were identified as Pseudomonas sp. and Psychrobacter sp. based on their 16S rDNA sequence. Crude phytase from these two bacterial strains was produced employing the shake flask method. Even though the total protein of the crude phytase was not significantly different for the two bacteria, the phytase activity of the crude enzyme produced by Pseudomonas sp. (97.1 ± 16.7 U) was significantly higher than that of the enzyme from Psychrobacter sp. (75.9 ± 2.4 U). When cod head kidney leukocytes were incubated with the crude phytase (50 μg ml−1), it resulted in enhanced cell proliferation, higher myeloperoxidase, and acid phosphatase activities. Extracellular responses—respiratory burst activity and hydrogen peroxide production were not enhanced by the crude enzyme. As a consequence, the growth of two pathogenic bacteria Aeromonas salmonicida and Vibrio anguillarum was not suppressed by the supernatants obtained from head kidney leukocytes incubated with the crude bacterial phytase. Thus, the enzyme from phytase-producing intestinal bacteria of Atlantic cod can stimulate intracellular head kidney leukocyte activities but not the production of extracellular substances that are involved in antibacterial response. These have implications on the potential use of bacterial phytase as feed supplement to boost cellular immune response of the fish and could be employed as a health management strategy in culture systems.

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
Fig. 6

Similar content being viewed by others

References

  • Alcorn SW, Pascho RJ, Murray AL, Shearer KD (2003) Effects of ration level on immune functions in Chinook salmon (Oncorhynchus tshawytscha). Aquaculture 217:529–545

    Article  Google Scholar 

  • Attwood EM, Weich DJ, Oosthuizen JM (1996) The influence of carbon particles on the concentration of acid phosphatase and lysozyme enzymes within alveolar macrophages during the killing and degradation of Mycobacterium bovis. Tuber Lung Dis 77:341–347

    Article  CAS  PubMed  Google Scholar 

  • Bianciotto V, Lumini E, Bonfante P, Vandame P (2003) ‘Candidatus Glomeribacter gigasporarum’ gen. nov., sp. Nov., an endosymbiont of arbuscular mycorrhizal fungi. Int J Syts Evol Microbiol 53:121–124

    Article  CAS  Google Scholar 

  • Budiño B, Cal RM, Piazzon MC, Lamas J (2006) The activity of several components of the innate immune system in diploid and triploid turbot. Comp Biochem Physiol A 125:108–113

    Google Scholar 

  • Caipang CMA, Hirono I, Aoki T (2003) Development of real-time PCR assay for the detection and quantification of red seabream iridovirus (RSIV). Fish Pathol 38:1–7

    CAS  Google Scholar 

  • Caipang CMA, Hirono I, Aoki T (2005) Induction of antiviral state in fish cells by Japanese flounder, Paralichthys olivaceus, interferon regulatory factor-1. Fish Shellfish Immunol 19:79–91

    Article  CAS  PubMed  Google Scholar 

  • Caipang CMA, Brinchmann MF, Kiron V (2008) Short-term overcrowding of Atlantic cod, Gadus morhua: effects on serum-mediated antibacterial activity and transcription of glucose transport and antioxidant defense related genes. Comp Biochem Physiol A 151:560–565

    Article  Google Scholar 

  • Cao L, Wang W, Yang C, Yang Y, Diana J, Yakupitiyage A, Lou Z, Li D (2007) Application of microbial phytase in fish feed. Enzyme Microbiol Tech 40:497–507

    Article  CAS  Google Scholar 

  • Cho CY, Bureau DP (2001) A review of diet formulation strategies and feeding systems to reduce excretory and feed wastes in aquaculture. Aquacult Res 32(Suppl 1):349–360

    CAS  Google Scholar 

  • Cho JS, Lee CW, Kang SH, Lee JC, Bok JD, Moon YS, Lee HG, Kim SC, Choi YJ (2003) Purification and characterization of a phytase from Pseudomonas syringae MOK1. Curr Microbiol 47:290–294

    Article  CAS  PubMed  Google Scholar 

  • Choct M (2006) Enzymes for the feed industry: past, present and future. W Poultry Sci J 62:5–16

  • Chung S, Secombes CJ (1988) Analysis of events occurring within teleost macrophage during the respiratory burst. Comp Biochem Physiol B 89:539–544

    Article  Google Scholar 

  • Dalsgaard J, Ekmann KS, Pedersen PB, Verlhac V (2009) Effect of supplemented fungal phytase on performance and phosphorus availability by phosphorus depleted juvenile rainbow trout (Oncorhynchus mykiss), and on the magnitude and composition of phosphorus waste output. Aquaculture 286:105–112

    Article  CAS  Google Scholar 

  • Dhanasiri AKS (2007) Changes in the gut flora of Atlantic cod (Gadus morhua) upon domestication. MS Thesis, Bodø University College, Norway 84 p

  • Greiner R, Farouk AE (2007) Purification and characterization of a bacterial phytase whose properties make it exceptionally useful as a feed supplement. Protein J 26:467–474

    Article  CAS  PubMed  Google Scholar 

  • Greiner R, Carlsson NG, Alminger ML (2000) Stereo-specificity of myo-inositol hexakisphosphate dephosphorylation by a phytate-degrading enzyme of Escherichia coli. J Biotechnol 84:53–62

    Article  CAS  Google Scholar 

  • Gulati HK, Chadha BS, Saini HS (2007) Production and characterization of thermostable alkaline phytase from Bacillus laevolacticus isolated from rhizophore soil. J Ind Microbiol Biotech 34:91–98

    Article  CAS  Google Scholar 

  • Hirimuthugoda NY, Chi Z, Li X, Wang L, Wu L (2006) Diversity of phytase producing marine yeasts. Ciencias Marinas 32:673–682

    Google Scholar 

  • Huang H, Luo H, Yang P, Meng K, Wang Y, Yuan T, Bai Y, Yao B (2006) A novel phytase with preferable characteristics from Yersinia intermedia. Biochem Biophys Res Commun 350:884–889

    Article  CAS  PubMed  Google Scholar 

  • In MJ, Jang ES, Kim YJ, Oh NS (2004) Purification and properties of an extracellular acid phytase from Pseudomonas fragi Y9451. J Microbiol Biotechnol 14:1004–1008

    CAS  Google Scholar 

  • Kim Y-H, Gwon M-N, Yang S-Y, Park T-K, Kim C-G, Kim C-W, Song M-D (2002) Isolation of phytase-producing Pseudomonas sp. and optimization of its phytase production. J Microbiol Biotech 12:279–285

    Google Scholar 

  • Klebanoff SJ, Clark RD (1978) The neutrophil: functions and clinical disorders. North Holland Publishing Company, Amsterdam, pp 409–466

    Google Scholar 

  • Li XY, Chi Z, Liu Z, Yan K, Li H (2008a) Phytase production by a marine yeast Kodamea ohmeri BG3. Appl Biochem Biotech 149:183–193

    Article  CAS  Google Scholar 

  • Li XY, Liu ZQ, Chi ZM (2008b) Production of phytase by a marine yeast Kodamaea ohmeri BG3 in an oats medium: optimization by response surface methodology. Biores Tech 99:6386–6390

    Article  CAS  Google Scholar 

  • Li XM, Chi Z, Liu Z, Li J, Wang X, Hirimuthugoda NS (2008c) Purification and characterization of extracellular phytase from a marine yeast Kodomaea ohmeri BG3. Marine Biotechnol 10:190–197

    Article  CAS  Google Scholar 

  • Liu N, Ru YJ, Cowieson AJ, Li FD, Cheng XCH (2008) Effects of phytate and phytase on the performance and immune function of broilers fed nutritionally marginal diets. Poultry Sci 87:1105–1111

    Article  CAS  Google Scholar 

  • Meng Z, Shao J, Xiang L (2003) CpG oligodeonucleotides activate grass carp (Ctenopharyngodon idellus) macrophages. Dev Comp Immunol 27:313–321

    Article  CAS  PubMed  Google Scholar 

  • Mondal S, Roy T, Sen SK, Ray AK (2008) Distribution of enzyme-producing bacteria in the digestive tracts of some freshwater fish. Acta Ichthyol et Piscat 38:1–8

    Article  Google Scholar 

  • Mukesh P, Suma S, Singaracharya MA, Lakshmipathi V (2004) Isolation of phytate hydrolysing microbial strains from traditional waste water of rice fermentation and liquid cattle feeds. World J Microbiol Biotech 20:531–534

    Article  CAS  Google Scholar 

  • Nakamura Y, Fukuhara H, Sano K (2000) Secreted phytase activities of yeasts. Biosci Biotech Biochem 64:841–844

    Article  CAS  Google Scholar 

  • Pavlova K, Gargova S, Hristozova T, Tankova Z (2008) Phytase from Antartic yeast strain Cryptococcus laurentii AL27. Folia Microbiol 53:29–34

    Article  CAS  Google Scholar 

  • Quade MJ, Roth JA (1997) A rapid, direct assay to measure degranulation of bovine neutrophil primary granules. Vet Immunol Immunopath 58:239–248

    Article  CAS  Google Scholar 

  • Richardson AE, Hadobas PA (1997) Soil isolates of Pseudomonas spp. that utilize inositol phosphates. Can J Microbiol 43:509–516

    Article  CAS  PubMed  Google Scholar 

  • Roy T, Mondal S, Ray AK (2008) Phytase-producing bacteria in the digestive tracts of some freshwater fish. Aquacult Res: 1–10

  • Saha S, Roy RN, Sen SK, Roy AK (2006) Characterization of cellulose-producing bacteria from the gastrointestinal tract of tilapia, Oreochromis mossambica (Peters) and grass carp, Ctenopharyngodon idella (Valenciennes). Aquacult Res 37:380–388

    Article  CAS  Google Scholar 

  • Sequeilha L, Lambrechts C, Boze H, Moulin G, Galzy P (2000) Purification and properties of the phytase from Schwanniomyces castellii. J Ferment Bioeng 74:7–11

    Article  Google Scholar 

  • Vats P, Banerjee UC (2005) Biochemical characterization of extracellular phytase (myo-inositol hexakisphosphate phosphohydrolase) from a hyper-producing strain of Aspergillus niger van Teighem. J Ind Microbiol Biotechnol 32:141–147

    Article  CAS  PubMed  Google Scholar 

  • Yoon SJ, Choi YJ, Min HK, Cho KK, Kim JW, Lee SC, Jung YH (1996) isolation and identification of phytase-producing bacterium, Enterobacter sp. 4, and enzymatic properties of phytase enzyme. Enzyme Microbiol Tech 18:449–454

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study is part of the Master’s thesis of the first author (C. L.) and was funded partly by a project of the Research Council of Norway “Mucosal immune system of Atlantic cod” (184703). The bacterial isolates used were obtained from a previous study of Anusha K.S. Dhanasiri to whom we are grateful. We also express our thanks to Ingvild Berg for providing the technical assistance.

Author information

Authors and Affiliations

  1. Faculty of Biosciences and Aquaculture, Bodø University College, 8049, Bodø, Norway

    Carlo C. Lazado, Christopher Marlowe A. Caipang, Sanchala Gallage, Monica F. Brinchmann & Viswanath Kiron

Authors
  1. Carlo C. Lazado
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christopher Marlowe A. Caipang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sanchala Gallage
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Monica F. Brinchmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Viswanath Kiron
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Christopher Marlowe A. Caipang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lazado, C.C., Caipang, C.M.A., Gallage, S. et al. Responses of Atlantic cod Gadus morhua head kidney leukocytes to phytase produced by gastrointestinal-derived bacteria. Fish Physiol Biochem 36, 883–891 (2010). https://doi.org/10.1007/s10695-009-9364-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10695-009-9364-0

Keywords

Search

Navigation