Fucoidan-rich Sargassum wightii extract supplemented with α-amylase improve growth and immune responses of Labeo rohita (Hamilton, 1822) fingerlings

  • K. A. Sajina
  • Narottam Prasad SahuEmail author
  • Tincy Varghese
  • Kamal Kant Jain


Fucoidan-rich seaweed extract (FRSE) was prepared from Sargassum wightii by modified alcohol-water extraction method. A 45-day feeding trial was conducted to evaluate the individual and combined effect of fucoidan-rich seaweed extract (FRSE—20 g kg−1) and exogenous α-amylase (100 mg kg−1) on the growth response, metabolic enzymes, immune parameters, and amylase gene expression of Labeo rohita. Four purified iso-nitrogenous diets (350 g CP kg−1 feed) with different combinations of α-amylase were prepared. Among the various treatment groups, the group fed with FRSE along with amylase group exhibited significantly higher (p < 0.05) weight gain, SGR, PER, liver metabolic enzyme activities, and lower FCR than the control group. The hematological and serum parameters such as NBT, erythrocyte count, plasma protein, albumin and globulin values were significantly higher (p < 0.05) in the FRSE fed groups. LDH activities and antioxidant enzyme activities reduced significantly in FRSE fed groups, whereas activities were significantly higher (p < 0.05) in the α-amylase fed and control groups. The intestinal amylase activities were significantly higher in the α-amylase fed groups, while expression of amylase gene mRNA was significantly lower in the α-amylase-supplemented groups (p < 0.05). There was no significant difference between control and FRSE fed groups in both, intestinal amylase activities and amylase gene mRNA expression. Overall results confirmed that FRSE extracted from S. wightii enhances immune responses, but not the growth. Further, it does not show any evidence of an inhibitory effect on intestinal amylase activity as well as on the mRNA levels. Hence, the α-amylase supplementation at 100 mg kg−1 with 20 g kg−1 FRSE in L. rohita fingerlings diet provides growth-promoting effect without compromising its immune-modulating effect.


Fucoidan rich seaweed extract Sargassum wightii α-Amylase Metabolic enzymes Labeo rohita 



This paper is a part of the M.Sc. thesis of the first author.

Funding information

Director, ICAR-Central Institute of Fisheries Education, Mumbai provided funding and the necessary facilities for the study. We hereby guarantee that the experiments and sampling of the fish comply with the existing norms of the institute and country.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Abasali H, Mohamad S (2010) Immune response of common carp (Cyprinus carpio) fed with herbal iminunostimulants diets. Asian J Anim Vet Adv 9:1839–1847CrossRefGoogle Scholar
  2. Adeoye AA, Jaramillo-Torres A, Fox SW, Merrifield DL, Davies SJ (2016) Supplementation of formulated diets for tilapia (Oreochromis niloticus) with selected exogenous enzymes: overall performance and effects on intestinal histology and microbiota. Anim Feed Sci Technol 215:133–143CrossRefGoogle Scholar
  3. Ale MT, Mikkelsen JD, Meyer AS (2011) Important determinants for fucoidan bioactivity: a critical review of structure-function relations and extraction methods for fucose-containing sulfated polysaccharides from brown seaweeds. Mar Drugs 9:2106–2130PubMedPubMedCentralCrossRefGoogle Scholar
  4. AOAC (1995) Official methods of analysis of the Association of Official Analytical Chemists (16th ed.). AOAC International, ArlingtonGoogle Scholar
  5. APHA-AWWA-WEF (1998) In: Clesceri LS, Greenberg AE, Eaton AD (eds) Standard methods for the examination of water and wastewater, 20th edn). American Public Health Association, American Water Works Association, Water Environment Federation, Washington DCGoogle Scholar
  6. Brand-Williams W, Cuvelier ME, Berset CLWT (1995) Use of a free radical method to evaluate antioxidant activity. LWT-Food Sci Technol 28:25–30CrossRefGoogle Scholar
  7. Bruhn A, Janicek T, Manns D, Nielsen MM, Balsby TJS, Meyer AS, Rasmussen MB, Hou X, Saake B, Göke C, Bjerre AB (2017) Crude fucoidan content in two North Atlantic kelp species, Saccharina latissima and Laminaria digitata—seasonal variation and impact of environmental factors. J Appl Phycol 29:3121–3137PubMedPubMedCentralCrossRefGoogle Scholar
  8. Cai Z, Li W, Mai K, Xu W, Zhang Y, Ai Q (2015) Effects of dietary size-fractionated fish hydrolysates on growth, activities of digestive enzymes and aminotransferases and expression of some protein metabolism related genes in large yellow croaker (Larimichthys crocea) larvae. Aquaculture 440:40–47CrossRefGoogle Scholar
  9. Chapman VJ, Chapman DJ (1980) Seaweeds and their uses, 4th edn. Chapman & Hall, London, pp 30–61Google Scholar
  10. Chatterjee N, Chen YH, Breslow NE (2003) A pseudoscore estimator for regression problems with two-phase sampling. J Am Stat Assoc 98:158–168CrossRefGoogle Scholar
  11. Cho M, Han JH, You S (2011) Inhibitory effects of fucan sulfates on enzymatic hydrolysis of starch. LWT-Food Sci Technol 44:1164–1171CrossRefGoogle Scholar
  12. Chotigeat W, Tongsupa S, Supamataya K, Phongdara A (2004) Effect of fucoidan on disease resistance of black tiger shrimp. Aquaculture 233:23–30CrossRefGoogle Scholar
  13. Cumashi A, Ushakova NA, Preobrazhenskaya ME, D'incecco A, Piccoli A, Totani L, Usov AI (2007) A comparative study of the anti-inflammatory, anticoagulant, antiangiogenic, and antiadhesive activities of nine different fucoidans from brown seaweeds. Glycobiology 17:541–552PubMedCrossRefGoogle Scholar
  14. De Jong J (2017) Aquaculture in India. Rijksdienstvoor Ondernemend Nederland (RVO. nl), NetherlandsGoogle Scholar
  15. Doner LW, Whistler RL (1973) Fucoidan: industrial gums, polysaccharides and their derivatives. Academic Press, New YorkCrossRefGoogle Scholar
  16. Doumas BT, Watson WA, Biggs HG (1971) Albumin standards and the measurement of serum albumin with bromcresol green. Clin Chim Acta 31:87–96PubMedCrossRefGoogle Scholar
  17. Drapeau G (1974) Protease from Staphylococcus aureus. In: Lorand BL (ed) Methods in Enzymology, vol 45. Academic Press, NY, p 469Google Scholar
  18. Dubois M, Gilles KA, Hamilton JK, Rebers PT, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356CrossRefGoogle Scholar
  19. Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42CrossRefGoogle Scholar
  20. Eluvakkal T, Sivakumar SR, Arunkumar K (2010) Fucoidan in some Indian brown seaweeds found along the coast gulf of Mannar. Int J Bot 6:176–181CrossRefGoogle Scholar
  21. FAO (Food and Agriculture Organization of United Nations) (2014). Fish to 2030. RomeGoogle Scholar
  22. Gora AH, Sahu NP, Sahoo S, Rehman S, Dar SA, Ahmad I, Agarwal D (2018) Effect of dietary Sargassum wightii and its fucoidan-rich extract on growth, immunity, disease resistance and antimicrobial peptide gene expression in Labeo rohita. Int Aquat Res:1–17Google Scholar
  23. Gupta S, Abu-Ghannam N (2011) Bioactive potential and possible health effects of edible brown seaweeds. Trends Food Sci Tech 22:315–326CrossRefGoogle Scholar
  24. Halver JE (1972) FAO technical conference on aquaculture: the nutritional requirement of cultivated warm water and cold water fish species. KyotoGoogle Scholar
  25. Han JG, Syed AQ, Kwon M, Ha JH, Lee HY (2008) Antioxident, immunomodulatory and anticancer activity of fucoidan isolated from Fucus vesiculosus. J Biotechnol 136:S571CrossRefGoogle Scholar
  26. Jiang Z, Zhou Y, Lu F, Han Z, Wang T (2008) Effects of different levels of supplementary alpha-amylase on digestive enzyme activities and pancreatic amylase mRNA expression of young broilers. Asian-Aust J Anim Sci 21:97–102CrossRefGoogle Scholar
  27. Jiménez-Escrig A, Gomez-Ordonez E, Ruperez P (2011) Seaweed as a source of novel nutraceuticals: sulfated polysaccharides and peptides. Adv Food Nutr Res 64:325–337Google Scholar
  28. Kaczmarek SA, Rogiewicz A, Mogielnicka M, Rutkowski A, Jones RO, Slominski BA (2014) The effect of protease, amylase, and nonstarch polysaccharide-degrading enzyme supplementation on nutrient utilization and growth performance of broiler chickens fed corn-soybean meal-based diets. Poult Sci 93:1745–1753PubMedCrossRefGoogle Scholar
  29. Kamalam BS, Medale F, Panserat S (2017) Utilisation of dietary carbohydrates in farmed fishes: new insights on influencing factors, biological limitations and future strategies. Aquaculture 467:3–27CrossRefGoogle Scholar
  30. Kim KJ, Yoon KY, Lee BY (2012) Low molecular weight fucoidan from the sporophyll of Undaria pinnatifida suppresses inflammation by promoting the inhibition of mitogen-activated protein kinases and oxidative stress in RAW264. 7 cells. Fitoterapia 83:1628–1635PubMedCrossRefGoogle Scholar
  31. Kim KT, Rioux LE, Turgeon SL (2014) Alpha-amylase and alpha-glucosidase inhibition is differentially modulated by fucoidan obtained from Fucus vesiculosus and Ascophyllum nodosum. Phytochemistry 98:27–33PubMedCrossRefGoogle Scholar
  32. Kim KT, Rioux LE, Turgeon SL (2015) Molecular weight and sulfate content modulate the inhibition of α-amylase by fucoidan relevant for type 2 diabetes management. Pharma Nutrition 3:108–114CrossRefGoogle Scholar
  33. Kohen R, Nyska A (2002) Oxidation of biological systems: oxidative stress phenomena, antioxidants, redox reactions, and methods for their quantification. Toxicol Pathol 30:620–650Google Scholar
  34. Koyanagi S, Tanigawa N, Nakagawa H, Soeda S, Shimeno H (2003) Oversulfation of fucoidan enhances its anti-angiogenic and antitumor activities. Biochem Pharmacol 65:173–179PubMedCrossRefGoogle Scholar
  35. Kumar S, Sahu NP, Pal AK, Choudhury D, Mukherjee SC (2006) Studies on digestibility and digestive enzyme activities in Labeo rohita (Hamilton) juveniles: effect of microbial α-amylase supplementation in non-gelatinized or gelatinized corn-based diet at two protein levels. Fish Physiol Biochem 32:209–220CrossRefGoogle Scholar
  36. Kumar TV, Lakshmanasenthil S, Geetharamani D, Marudhupandi T, Suja G, Suganya P (2015) Fucoidan—a α-d-glucosidase inhibitor from Sargassum wightii with relevance to type 2 diabetes mellitus therapy. Int J Biol Macromol 72:1044–1047CrossRefGoogle Scholar
  37. Lakshmanasenthil S, Vinothkumar T, Geetharamani D, Marudhupandi T, Suja G, Sindhu NS (2014) Fucoidan—a novel α-amylase inhibitor from Turbinaria ornata with relevance to NIDDM therapy. Biocatal Agric Biotechnol 3:66–70CrossRefGoogle Scholar
  38. Lee SH, Ko CI, Jee Y, Jeong Y, Kim M, Kim JS, Jeon YJ (2013) Anti-inflammatory effect of fucoidan extracted from Ecklonia cava in zebrafish model. Carbohyd Polym 92:84–89CrossRefGoogle Scholar
  39. Li B, Lu F, Wei X, Zhao R (2008) Fucoidan: structure and bioactivity. Molecules 13:1671–1695PubMedPubMedCentralCrossRefGoogle Scholar
  40. Lin S, Mai K, Tan B (2007) Effects of exogenous enzyme supplementation in diets on growth and feed utilization in tilapia, Oreochromis niloticus x O. aureus. Aquac Res 38:1645–1653CrossRefGoogle Scholar
  41. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408CrossRefGoogle Scholar
  42. Mabeau S, Fleurence J (1993) Seaweed in food products: biochemical and nutritional aspects. Trends Food Sci Technol 4:103–107CrossRefGoogle Scholar
  43. Maki KC, Galant R, Samuel P, Tesser J, Witchger MS, Ribaya-Mercado JD, Geohas J (2007) Effects of consuming foods containing oat β-glucan on blood pressure, carbohydrate metabolism and biomarkers of oxidative stress in men and women with elevated blood pressure. Eur J Clin Nutr 61:786–795PubMedCrossRefGoogle Scholar
  44. Mir IN, Sahu NP, Pal AK, Makesh M (2017) Synergistic effect of l-methionine and fucoidan rich extract in eliciting growth and non-specific immune response of Labeo rohita fingerlings against Aeromonas hydrophila. Aquaculture 479:396–403CrossRefGoogle Scholar
  45. Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170–3175PubMedGoogle Scholar
  46. Ortiz J, Romero N, Robert P, Araya J, Lopez-Hernández J, Bozzo C, Navarrete E, Osorio A, Rios A (2006) Dietary fiber, amino acid, fatty acid and tocopherol contents of the edible seaweeds Ulva lactuca and Durvillaea antarctica. Food Chem 99:98–104CrossRefGoogle Scholar
  47. Ou S, Kwok KC, Li Y, Fu L (2001) In vitro study of possible role of dietary fiber in lowering postprandial serum glucose. J Agric Food Chem 49:1026–1029PubMedCrossRefGoogle Scholar
  48. Ponce NM, Pujol CA, Damonte EB, Flores ML, Stortz CA (2003) Fucoidans from the brown seaweed Adenocystis utricularis: extraction methods, antiviral activity and structural studies. Carbohydr Res 338:153–165PubMedCrossRefGoogle Scholar
  49. Prabu DL, Sahu NP, Pal AK, Dasgupta S, Narendra A (2016) Immunomodulation and interferon gamma gene expression in sutchi cat fish, Pangasianodon hypophthalmus: effect of dietary fucoidan rich seaweed extract (FRSE) on pre and post challenge period. Aquac Res 47:199–218CrossRefGoogle Scholar
  50. Reinhold JG (1953) Total protein, albumin and globulin. In: Standard methods of clinical chemistry, Academic Press, NY, Vol 1. S88Google Scholar
  51. Rick W, Stegbauer HP (1974) Alpha amylase measurement of reducing groups. In: Methods of enzymetic analysis, 2nd edn. Academic Press, New DehliGoogle Scholar
  52. Shan X, Liu X, Hao J, Cai C, Fan F, Dun Y, ZhaoX LX, Li C, Yu G (2016) In vitro and in vivo hypoglycemic effects of brown algal fucoidans. Int J Biol Macromol 82:249–255PubMedCrossRefGoogle Scholar
  53. Shanura Fernando IP, Asanka Sanjeewa KK, Samarakoon KW, Kim H-S, Gunasekara UKDSS, Park Y-J, Abeytunga DTU, Lee WW, Jeon Y-J (2018) The potential of fucoidans from Chnoospora minima and Sargassum polycystum in cosmetics: antioxidant, anti-inflammatory, skin-whitening, and antiwrinkle activities. J Appl Phycol 30:3223–3232CrossRefGoogle Scholar
  54. Singleton VL, Rossi JA (1965) Colorimetry of total phenolics with phosphomolybdic-phospho tungstic acid reagents. Am J Enol Viticult 16:144–158Google Scholar
  55. Stasiack AS, Bauman CP (1996) Netrophil activity as a potential bioindicator for containment analysis. Fish Shellfish Immunol 6:537–539Google Scholar
  56. Takahara S, Hamilton HB, Neel JV, Kobara TY, Ogura Y, Nishimura ET (1960) Hypocatalasemia: a new genetic carrier state. J Clin Inv 39:610–619CrossRefGoogle Scholar
  57. Traifalgar RF, Kira H, Thanh Tung H, Raafat Michael F, Laining A, Yokoyama S, IShikawa M, Koshio S, Serrano AE, Corre V (2010) Influence of dietary fucoidan supplementation on growth and immunological response of juvenile Marsupenaeus japonicus. J World Aquacult Soc 41:235–244CrossRefGoogle Scholar
  58. Tuller J, De Santis C, Jerry DR (2014) Dietary influence of Fucoidan supplementation on growth of Lates calcarifer (Bloch). Aquac Res 45:749–754CrossRefGoogle Scholar
  59. Van-Der-Maarel MJ, Van der Veen B, Uitdehaag JC, Leemhuis H, Dijkhuizen L (2002) Properties and applications of starch-converting enzymes of the α-amylase family. J Biotech 94:137–155CrossRefGoogle Scholar
  60. Vo TS, Kim SK (2013) Fucoidans as a natural bioactive ingredient for functional foods. J Funct Foods 5:16–27CrossRefGoogle Scholar
  61. Wang J, Zhang Q, Zhang Z, Li Z (2008) Antioxidant activity of sulfated polysaccharide fractions extracted from Laminaria japonica. Int J Biol Macromol 42:127–132PubMedCrossRefGoogle Scholar
  62. Weeks BA, Warinner JE, Mason PL, McGinnis DS (1986) Influence of toxic chemicals on the chemotactic response of fish macrophages. J Fish Biol 28:653–658CrossRefGoogle Scholar
  63. Wooten IDP (1964) Microanalysis. In: Medical Biochemistry (4thed.) LondonGoogle Scholar
  64. Wroblewski F, Laude JS (1955) LDH activity in blood. Proc Soc Exp Biol Med 90:210–213PubMedCrossRefGoogle Scholar
  65. Xu Y, Zhang Q, Luo D, Wang J, Duan D (2017) Low molecular weight fucoidan ameliorates the inflammation and glomerular filtration function of diabetic nephropathy. J Appl Phycol 29:531–542CrossRefGoogle Scholar
  66. Yang C, Chung D, You S (2008) Determination of physicochemical properties of sulphated fucans from sporophyll of Undaria pinnatifida using light scattering technique. Food Chem 111:503–507PubMedCrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • K. A. Sajina
    • 1
  • Narottam Prasad Sahu
    • 1
    Email author
  • Tincy Varghese
    • 1
  • Kamal Kant Jain
    • 1
  1. 1.ICAR-Central Institute of Fisheries Education (CIFE)MumbaiIndia

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