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Nucleotide Enrichment of Live Feed: A Promising Protocol for Rearing of Atlantic Cod Gadus morhua Larvae

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Abstract

We investigated the effect of two commercial nucleotide products (NT1 and NT2), administered through live feed, on growth and stress tolerance of Atlantic cod larvae. Expression of genes related to muscle growth (igf-1, igf1r, igf-2, fst, fgf6, myod, and myhc) and nucleotide metabolism (uox, hprt, ndk, and uck) was evaluated during larval development. In addition, the expression of genes related to stress (hif-1α, hif-2α, hif-3α, and mb) was studied after an air exposure stress test. The enrichment of rotifers with nucleotides did not reveal any difference in nucleotide profiles, the exception being the RNA level of the NT1-enriched group that was significantly higher than the unenriched rotifer. Unenriched Artemia showed poor nucleotide profiles compared to enriched Artemia since 5′ UMP, 5′ GMP, and 5′ AMP were observed only in the nucleotide groups. At 38 days post-hatch (dph), NT1 group had significantly higher dry weight (3.1 ± 0.1 mg) than the control (CON; 2.3 ± 0.1 mg). The treatments did not produce any significant differences in the expression of the key myogenic genes. Among the genes associated with nucleotide metabolism, ndk was down-regulated in NT1 at 38 dph. In the air exposure test, survival was significantly higher in the CON (77 ± 6 %) than in NT1 (48 ± 3 %) and NT2 (50 ± 3 %). After air exposure, mb was expressed at lower levels in NT2 group, hif-2α was induced in NT1 group, and hif-3α was upregulated in all groups. Our findings indicate that the improvement in the nucleotide profile of Artemia upon nucleotide enrichment could eventuate in the rapid growth of larvae.

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References

  • Alami-Durante H, Médale F, Cluzeaud M, Kaushik SJ (2010a) Skeletal muscle growth dynamics and expression of related genes in white and red muscles of rainbow trout fed diets with graded levels of a mixture of plant protein sources as substitutes for fishmeal. Aquaculture 303:50–58

    Article  CAS  Google Scholar 

  • Alami-Durante H, Wrutniak-Cabello C, Kaushik SJ, Médale F (2010b) Skeletal muscle cellularity and expression of myogenic regulatory factors and myosin heavy chains in rainbow trout (Oncorhynchus mykiss): effects of changes in dietary plant protein sources and amino acid profiles. Comp Biochem Physiol A 156:561–568

    Article  CAS  Google Scholar 

  • Andersen Ø, Aas TS, Skugor S, Takle H, van Nes S, Grisdale-Helland B, Helland SJ, Terjesen BF (2006) Purine-induced expression of urate oxidase and enzyme activity in Atlantic salmon (Salmo salar). FEBS J 273:2839–2850

    Article  PubMed  CAS  Google Scholar 

  • Ayson FG, de Jesus EG, Moriyama S, Hyodo S, Funkenstein B, Gertler A, Kawauchi H (2002) Differential expression of insulin-like growth factor I and II mRNAs during embryogenesis and early larval development in rabbitfish, Siganus guttatus. Gen Comp Endocrinol 126:165–174

    Article  PubMed  CAS  Google Scholar 

  • Baskerville-Bridges B, Kling LJ (2000) Early weaning of Atlantic cod (Gadus morhua) larvae onto a microparticulate diet. Aquaculture 189:109–117

    Article  Google Scholar 

  • Burrells C, Williams PD, Southgate PJ, Wadsworth SL (2001) Dietary nucleotides: a novel supplement in fish feeds: 2. Effects on vaccination, salt water transfer, growth rates and physiology of Atlantic salmon (Salmo salar L.). Aquaculture 199:171–184

    Article  CAS  Google Scholar 

  • Busch KET, Falk-Petersen IB, Peruzzi S, Rist NA, Hamre K (2010) Natural zooplankton as larval feed in intensive rearing systems for juvenile production of Atlantic cod (Gadus morhua L.). Aquac Res 41:1727–1740

    Article  Google Scholar 

  • Campos C, Valente LMP, Borges P, Bizuayehu T, Fernandes JMO (2010) Dietary lipid levels have a remarkable impact on the expression of growth-related genes in Senegalese sole (Solea senegalensis Kaup). J Exp Biol 213:200–209

    Article  PubMed  CAS  Google Scholar 

  • Carver JD (1999) Dietary nucleotides: effects on the immune and gastrointestinal systems. Acta Paediatr Suppl 88:83–88

    Article  PubMed  CAS  Google Scholar 

  • Chapalamadugu KC, Robison BD, Drew RE, Powell MS, Hill RA, Amberg JJ, Rodnick KJ, Hardy RW, Hill ML, Murdoch GK (2009) Dietary carbohydrate level affects transcription factor expression that regulates skeletal muscle myogenesis in rainbow trout. Comp Biochem Physiol B 153:66–72

    Article  PubMed  Google Scholar 

  • Cheng Z, Buentello A, Gatlin DM III (2011) Dietary nucleotides influence immune responses and intestinal morphology of red drum Sciaenops ocellatus. Fish Shellfish Immunol 30:143–147

    Article  PubMed  CAS  Google Scholar 

  • Conceição LEC, Yúfera M, Makridis P, Morais S, Dinis MT (2010) Live feeds for early stages of fish rearing. Aquac Res 41:613–640

    Article  Google Scholar 

  • Cosgrove M (1998) Nucleotides. Nutrition 14:748–751

    Article  PubMed  CAS  Google Scholar 

  • De-Santis C, Jerry DR (2007) Candidate growth genes in finfish—where should we be looking? Aquaculture 272:22–38

    Article  CAS  Google Scholar 

  • Eliès G, Duval H, Bonnec G, Wolff J, Boeuf G, Boujard D (1999) Insulin and insulin-like growth factor-1 receptors in an evoluted fish, the turbot: cDNA cloning and mRNA expression. Mol Cell Endocrinol 158:173–185

    Article  PubMed  Google Scholar 

  • Falk-Petersen IB (2005) Comparative organ differentiation during early life stages of marine fish. Fish Shellfish Immunol 19:397–412

    Article  PubMed  CAS  Google Scholar 

  • Fernandes JMO, Kinghorn JR, Johnston IA (2007) Differential regulation of multiple alternatively spliced transcripts of MyoD. Gene 391:178–185

    Article  PubMed  CAS  Google Scholar 

  • Fernandes JMO, Mommens M, Hagen Ø, Babiak I, Solberg C (2008) Selection of suitable reference genes for real-time PCR studies of Atlantic halibut development. Comp Biochem Physiol B 150:23–32

    Article  PubMed  Google Scholar 

  • Florini JR, Ewton DZ, Coolican SA (1996) Growth hormone and the insulin-like growth factor system in myogenesis. Endocr Rev 17:481–517

    PubMed  CAS  Google Scholar 

  • Funes V, Asensio E, Ponce M, Infante C, Canavate JP, Manchado M (2006) Insulin-like growth factors I and II in the sole Solea senegalensis: cDNA cloning and quantitation of gene expression in tissues and during larval development. Gen Comp Endocrinol 149:166–172

    Article  PubMed  CAS  Google Scholar 

  • Funkenstein B, Rebhan Y, Skopal T (2009) Molecular cloning and characterization of follistatin in the gilthead sea bream, Sparus aurata. Mol Biol Rep 36:501–511

    Article  PubMed  CAS  Google Scholar 

  • Galloway TF, Kjørsvik E, Kryvi H (1999) Muscle growth and development in Atlantic cod larvae (Gadus morhua L.), related to different somatic growth rates. J Exp Biol 202:2111–2120

    PubMed  CAS  Google Scholar 

  • Gauvry L, Fauconneau B (1996) Cloning of a trout fast skeletal myosin heavy chain expressed both in embryo and adult muscles and in myotubes neoformed in vitro. Comp Biochem Physiol B 115:183–190

    Article  PubMed  CAS  Google Scholar 

  • Greene MW, Chen TT (1999) Quantitation of IGF-I, IGF-II, and multiple insulin receptor family member messenger RNAs during embryonic development in rainbow trout. Mol Reprod Dev 54:348–361

    Article  PubMed  CAS  Google Scholar 

  • Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  • Hall TE, Cole NJ, Johnston IA (2003) Temperature and the expression of seven muscle-specific protein genes during embryogenesis in the Atlantic cod Gadus morhua L. J Exp Biol 206:3187–3200

    Article  PubMed  CAS  Google Scholar 

  • Heidbreder M, Fröhlich F, Jöhren O, Dendorfer A, Qadri F, Dominiak P (2003) Hypoxia rapidly activates HIF-3alpha mRNA expression. FASEB J 17:1541–1543

    PubMed  CAS  Google Scholar 

  • Hevrøy EM, Jordal A-EO, Hordvik I, Espe M, Hemre G-I, Olsvik PA (2006) Myosin heavy chain mRNA expression correlates higher with muscle protein accretion than growth in Atlantic salmon, Salmo salar. Aquaculture 252:453–461

    Article  Google Scholar 

  • Hill JJ, Davies MV, Pearson AA, Wang JH, Hewick RM, Wolfman NM, Qiu Y (2002) The myostatin propeptide and the follistatin-related gene are inhibitory binding proteins of myostatin in normal serum. J Biol Chem 277:40735–40741

    Article  PubMed  CAS  Google Scholar 

  • Hiroi J, Sakakura Y, Tagawa M, Seikai T, Tanaka M (1997) Developmental changes in low-salinity tolerance and responses of prolactin, cortisol and thyroid hormones to low-salinity environment in larvae and juveniles of Japanese flounder, Paralichthys olivaceus. Zool Sci 14:987–992

    Article  CAS  Google Scholar 

  • Jiralerspong S, Patel PI (1996) Regulation of the hypoxanthine phosphoribosyltransferase gene: in vitro and in vivo approaches. Proc Soc Exp Biol Med 212:116–127

    PubMed  CAS  Google Scholar 

  • Johansen KA, Overturf K (2005) Quantitative expression analysis of genes affecting muscle growth during development of rainbow trout (Oncorhynchus mykiss). Mar Biotechnol 7:576–587

    Article  PubMed  CAS  Google Scholar 

  • Keyvanshokooh S, Tahmasebi-Kohyani A (2012) Proteome modifications of fingerling rainbow trout (Oncorhynchus mykiss) muscle as an effect of dietary nucleotides. Aquaculture 324–325:79–84

    Article  Google Scholar 

  • King VW, Berlinsky DL (2006) Whole-body corticosteroid and plasma cortisol concentrations in larval and juvenile Atlantic cod Gadus morhua L. following acute stress. Aquac Res 37:1282–1289

    Article  CAS  Google Scholar 

  • Kortner TM, Overrein I, Øie G, Kjørsvik E, Arukwe A (2011) The influence of dietary constituents on the molecular ontogeny of digestive capability and effects on growth and appetite in Atlantic cod larvae (Gadus morhua). Aquaculture 315:114–120

    Article  CAS  Google Scholar 

  • Law SHW, Wu RSS, Ng PKS, Yu RMK, Kong RYC (2006) Cloning and expression analysis of two distinct HIF-alpha isoforms—gcHIF-1alpha and gcHIF-4alpha—from the hypoxia-tolerant grass carp. Ctenopharyngodon idellus. BMC Mol Biol 7:15

    Article  PubMed  Google Scholar 

  • Li P, Gatlin DM III (2006) Nucleotide nutrition in fish: current knowledge and future applications. Aquaculture 251:141–152

    Article  CAS  Google Scholar 

  • Li P, Burr GS, Goff J, Whiteman KW, Davis KB, Vega RR, Neill WH, Gatlin DM III (2005) A preliminary study on the effects of dietary supplementation of brewers yeast and nucleotides, singularly or in combination, on juvenile red drum (Sciaenops ocellatus). Aquac Res 36:1120–1127

    Article  CAS  Google Scholar 

  • Lin YH, Wang H, Shiau SY (2009) Dietary nucleotide supplementation enhances growth and immune responses of grouper, Epinephelus malabaricus. Aquac Nutr 15:117–122

    Article  CAS  Google Scholar 

  • Lopez-Navarro AT, Ortega MA, Peragón J, Bueno JD, Gil A, Sánchez-Pozo A (1996) Deprivation of dietary nucleotides decreases protein synthesis in the liver and small intestine in rats. Gastroenterology 110:1760–1769

    Article  PubMed  CAS  Google Scholar 

  • Maloney D (1998) Yeasts. In: Kroschwitz JI, Howe-Grant M (eds) Kirk–Othmer encyclopedia of chemical technology, 4th edn. Wiley, New York, pp 761–788

    Google Scholar 

  • McCollum A, Geubtner J, von Herbing IH (2006) Metabolic cost of feeding in Atlantic cod (Gadus morhua) larvae using microcalorimetry. ICES J Mar Sci 63:335–339

    Article  Google Scholar 

  • Peterson BC, Bosworth BG, Bilodeau AL (2005) Differential gene expression of IGF-I, IGF-II, and toll-like receptors 3 and 5 during embryogenesis in hybrid (channel × blue) and channel catfish. Comp Biochem Physiol A 141:42–47

    Article  Google Scholar 

  • Ponce M, Infante C, Funes V, Manchado M (2008) Molecular characterization and gene expression analysis of insulin-like growth factors I and II in the redbanded seabream, Pagrus auriga: transcriptional regulation by growth hormone. Comp Biochem Physiol B 150:418–426

    Article  PubMed  Google Scholar 

  • Rahman MS, Thomas P (2007) Molecular cloning, characterization and expression of two hypoxia-inducible factor alpha subunits, HIF-1alpha and HIF-2alpha, in a hypoxia-tolerant marine teleost, Atlantic croaker (Micropogonias undulatus). Gene 396:273–282

    Article  PubMed  CAS  Google Scholar 

  • Rescan PY (1998) Identification of a fibroblast growth factor 6 (FGF6) gene in a non-mammalian vertebrate: continuous expression of FGF6 accompanies muscle fiber hyperplasia. Biochim Biophys Acta 1443:305–314

    Article  PubMed  CAS  Google Scholar 

  • Rescan PY (2005) Muscle growth patterns and regulation during fish ontogeny. Gen Comp Endocrinol 142:111–116

    Article  PubMed  CAS  Google Scholar 

  • Ricker WE (1958) Handbook of computations for biological statistics of fish populations. Bull Fish Res Board Can 119:1–30

    Google Scholar 

  • Sæle Ø, Nordgreen A, Hamre K, Olsvik PA (2009) Evaluation of candidate reference genes in Q-PCR studies of Atlantic cod (Gadus morhua) ontogeny, with emphasis on the gastrointestinal tract. Comp Biochem Physiol B 152:94–101

    Article  PubMed  Google Scholar 

  • Sánchez-Pozo A, Gil A (2002) Nucleotides as semi-essential nutritional components. Br J Nutr 87:S135–S137

    Article  PubMed  Google Scholar 

  • Sauer N, Mosenthin R, Bauer E (2011) The role of dietary nucleotides in single-stomached animals. Nutr Res Rev 24:46–59

    Article  CAS  Google Scholar 

  • Shen R-J, Jiang X-Y, Pu J-W, Zou S-M (2010) HIF-1α and -2α genes in a hypoxia-sensitive teleost species Megalobrama amblycephala: cDNA cloning, expression and different responses to hypoxia. Comp Biochem Physiol B 157:273–280

    Article  PubMed  Google Scholar 

  • Soitamo AJ, Råbergh CMI, Gassmann M, Sistonen L, Nikinmaa M (2001) Characterization of a hypoxia-inducible factor (HIF-1α) from rainbow trout. J Biol Chem 276:19699–19705

    Article  PubMed  CAS  Google Scholar 

  • Tahmasebi-Kohyani A, Keyvanshokooh S, Nematollahi A, Mahmoudi N, Pasha-Zanoosi H (2011) Dietary administration of nucleotides to enhance growth, humoral immune responses, and disease resistance of the rainbow trout (Oncorhynchus mykiss) fingerlings. Fish Shellfish Immunol 30:189–193

    Article  PubMed  CAS  Google Scholar 

  • Terova G, Rimoldi S, Corà S, Bernadini G, Gornati R, Saroglia M (2008) Acute and chronic hypoxia affects HIF-1α mRNA levels in sea bass (Dicentrarchus labrax). Aquaculture 279:150–159

    Article  CAS  Google Scholar 

  • Uauy R, Quan R, Gil A (1994) Role of nucleotides in intestinal development and repair: implications for infant nutrition. J Nutr 124:1436S–1441S

    PubMed  CAS  Google Scholar 

  • Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3:RESEARCH0034

    Article  PubMed  Google Scholar 

  • Walsh MJ, Sanchez-Pozo A, Leleiko NS (1990) A regulatory element is characterized by purine-mediated and cell-type-specific gene transcription. Mol Cell Biol 10:4536–4364

    Google Scholar 

  • Walsh MJ, Tsao K-L, Leleiko NS (1992) Characterization of DNA–protein interactions within a distal regulatory element upstream of a mammalian housekeeping gene promoter. J Biol Chem 267:7026–7035

    PubMed  CAS  Google Scholar 

  • Welker TL, Lim C, Yildirim-Aksoy M, Klesius PH (2011) Effects of dietary supplementation of a purified nucleotide mixture on immune function and disease and stress resistance in channel catfish, Ictalurus punctatus. Aquac Res 42:1878–1889

    Article  CAS  Google Scholar 

  • Welt C, Sidis Y, Keutmann H, Schneyer A (2002) Activins, inhibins, and follistatins: from endocrinology to signaling. A paradigm for the new millennium. Exp Biol Med 227:724–752

    CAS  Google Scholar 

  • Werner A (1991) Analysis of nucleotides, nucleosides, nucleobases in cells by ion-pair reversed-phase HPLC. Chromatographia 31:401–410

    Article  CAS  Google Scholar 

  • Yousefi M, Abtahi B, Kenari AA (2011) Hematological, serum biochemical parameters, and physiological responses to acute stress of Beluga sturgeon (Huso huso, Linnaeus 1785) juveniles fed dietary nucleotide. Comp Clin Path 1-6. doi:10.1007/00580-011-1225-4

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Acknowledgments

The study was funded by the Research Council of Norway, project 182653/V10. The authors appreciate the excellent technical assistance provided by Bjørnar Eggen, Magnus Røkke, and Heidi Ludviksen during live feed production and collection of samples. The support of Rudi Ripman Seim and Odd Elvebø from Biorigin Scandinavia AS, Oslo, Norway, is gratefully acknowledged. Peter Koeppel, Chemoforma Ltd, Augst, Switzerland, is thanked for gifting one of the nucleotide products. The nucleotide analysis of the live feeds was kindly performed by Biorigin Brazil A/S, São Paulo, Brazil. Norwegian Cod Breeding Centre, Tromsø, Norway, supplied the eggs for the study. C.F.C. Lanes is a Ph.D. student supported by funds from Research Council of Norway and University of Nordland.

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

    Carlos Frederico Ceccon Lanes, Sylvie Bolla, Jorge M. O. Fernandes, Ove Nicolaisen, Viswanath Kiron & Igor Babiak

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  1. Carlos Frederico Ceccon Lanes
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  2. Sylvie Bolla
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  3. Jorge M. O. Fernandes
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Correspondence to Viswanath Kiron.

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Lanes, C.F.C., Bolla, S., Fernandes, J.M.O. et al. Nucleotide Enrichment of Live Feed: A Promising Protocol for Rearing of Atlantic Cod Gadus morhua Larvae. Mar Biotechnol 14, 544–558 (2012). https://doi.org/10.1007/s10126-012-9458-z

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