Skip to main content

Advertisement

SpringerLink
Log in

Screening and characterization of antiviral compounds from Enteromorpha flexuosa against white spot syndrome virus (WSSV) and its in vivo influence on Indian white shrimp Fenneropenaeus indicus

  • Published:
Aquaculture International Aims and scope Submit manuscript

Abstract

In vitro antiviral activities of the organic solvent extracts of six species of seaweeds collected from the Kanyakumari Coast, India, were screened against shrimp white spot syndrome virus (WSSV). Based on the primary screening, ethyl acetate extract of Enteromorpha flexuosa effectively suppressed WSSV multiplication, and this extract was further purified through silica column chromatography and resulted in different fractions. After the secondary antiviral screening against WSSV, fraction III (FIII) effectively controlled WSSV multiplication at the 95 % level, which was detected by two-step PCR. Thin-layer chromatography and GCMS analysis revealed that the phenolic antiviral compound 2-(2-hydroxyphenoxy)-1-phenylethanol is present in the FIII fraction. Further, the active fraction FIII was incorporated into the artificial diets at different concentrations of 100 (D1), 200 (D2), 300 (D3) and 400 (D4) mg kg−1. A control diet, devoid of seaweed extract, was also prepared. The control and experimental diets were fed to the Indian white shrimp Fenneropenaeus indicus for 25 days; they were then challenged with virulent WSSV. The shrimp control group was 100 % PCR positive, whereas the D4 group was only 5 % positive after two-step PCR detection, and the value differed significantly (F = 7.31; P ≤ 0.05). The biochemical parameters of the shrimp groups fed with experimental diets significantly improved (P < 0.001) the coagualse activity to 94.54 s and increased the oxyhemocyanin level to 1.12 m mol l−1. The immunological parameters such as prophenoloxidase and lysozyme activity also appeared to be significantly increased (F = 164.88; P ≤ 0.001) in the experimental compared to control groups.

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

  • Anand Ganesh E, Das S, Arun G, Balamurugan S, Raj RR (2009) Heparin like compound from green alga Chaetomorpha antennina as potential anticoagulant agent. Asian J Med Sci 1(3):114–116

    Google Scholar 

  • Armstrong FAJ, Boalch GT (1960) Volatile organic matter in algal culture media and in seawater. Nature 185:761–762

    Article  CAS  Google Scholar 

  • Athukorala Y, Jung WK, Vasanthan T, Jeon YJ (2006) An anticoagulative polysaccharide from an enzymatic hydrolysate of Ecklonia cava. Carbohydr Polym 66:184–191

    Article  CAS  Google Scholar 

  • Balasubramanian G, Sarathi M, Venkatesan C, Thomas J, Sahul Hameed AS (2008) Oral administration of antiviral plant extract of Cynodon dactylon on a large scale production against White spot syndrome virus (WSSV) in Penaeus monodon. Aquaculture 279:2–5

  • Boonyaratpalin M (1993) Nutritional requirements of grouper Epinephelus. The proceedings of Grouper culture. National Institute of coastal Aquaculture. Department of fisheries, Thailand, pp 50–55

  • Chang CF, Su MS, Chen HY, Lo CF, Kou GH, Liao IC (1999) Effect of dietary β 1,3-glucan on resistance to white spot syndrome virus (WSSV) in postlarval and juvenile Penaeus monodon. Dis Aquat Organ 36:163–168

    Article  CAS  Google Scholar 

  • Chattopadhyay K, Mandal P, Lerouge P, Driouich A, Ghosal P, Ray B (2007) Sulphated polysaccharides from Indian samples of Enteromorpha compressa (Ulvales, Chlorophyta): isolation and structural features. Food Chem 104:928–935

    Article  CAS  Google Scholar 

  • Chotigeat W, Tongsupa S, Supamataya K, Phongdara A (2004) Effect of fucoidan on disease resistance of black tiger shrimp. Aquaculture 233:23–30

    Article  CAS  Google Scholar 

  • Citarasu T, Sivaram V, Immanuel G, Rout N, Murugan V (2006) Influence of selected Indian immunostimulant herbs against White Spot Syndrome Virus (WSSV) infection in black tiger shrimp, Penaeus monodon with reference to haematological, biochemical and immunological changes. Fish Shellfish Immunol 21:372–384

    Article  PubMed  Google Scholar 

  • Craigie JS, Gruenig DE (1967) Bromophenols from red algae. Science 157:1058–1059

    Article  CAS  PubMed  Google Scholar 

  • FAO (2009) The State of World Fisheries and Aquaculture—2008 (SOFIA). FAO Fisheries and Aquaculture Department, p 176

  • FAO (2012) The State of World Fisheries and Aquaculture. Food and Agriculture Organization of the United Nations, Viale delle Terme di Caracalla, 00153 Rome, Italy

  • Feldman SC, Reynaldi S, Stortz CA, Cerezo AS, Damonte EB (1999) Antiviral properties of fucoidan fractions from Leathesia difformis. Phytomedicine 6(5):335–340

    Article  CAS  PubMed  Google Scholar 

  • Fenical W, Gifkins KB, Clardy J (1974) X-ray determination of chondriol: a re-assignment of structure. Tetrahedron Lett 15:1507–1510

    Article  Google Scholar 

  • Hass P (1935) The liberation of methyl sulfoxide by seaweed. Biochem J 29:1297–1299

    Google Scholar 

  • Heo SJ, Park EJ, Lee KW, Jeon YJ (2005) Antioxidant activities of enzymatic extracts from brown seaweeds. Bioresource Technol 96:1613–1623

  • Hodgkin JH, Craigie JS, McInnes AG (1966) The occurrence of 2,3-dibrombenzyl alcohol-4,5-disulfate dipotassium salt in Polysiphonia lanosa. Can J Chem 44:74–78

    Article  CAS  Google Scholar 

  • Huang X, Zhou H, Zhang H (2006) The effect of Sargassum fusiforme polysaccharide extracts on vibriosis resistance and immune activity of the shrimp, Fenneropenaeus chinensis. Fish Shellfish Immunol 20:750–757

    Article  CAS  PubMed  Google Scholar 

  • Huynh TG, Yeh ST, Lin YC, Shyu JF, Chen LL, Chen JC (2011) White shrimp Litopenaeus vannamei immersed in seawater containing Sargassum hemiphyllum var. chinense powder and its extract showed increased immunity and resistance against Vibrio alginolyticus and white spot syndrome virus. Fish Shellfish Immunol 31:286–293

    Article  CAS  PubMed  Google Scholar 

  • Ivanova V, Rouseva R, Kolarova M, Serkedjieva J, Rachev R, Manolova N (1994) Isolation of a Polysaccharide with Antiviral Effect from Ulva lactuca. Prep Biochem 24:83–97

  • Johansson MW, Keyser P, Sritunyaluksana K, Söderhäll K (2000) Crustacean hemocytes and haematopoiesis. Aquaculture 191:45–52

    Article  CAS  Google Scholar 

  • Kim TH, Bae JS (2010) Ecklonia cava extracts inhibit liposaccharide induced inflammatory responses in human endothelial cells. Food Chem Toxicol 48:1682–1687

    Article  CAS  PubMed  Google Scholar 

  • Kim KJ, Lee OH, Lee BY (2010) Fucoidan, a sulfated polysaccharide, inhibits adipogenesis through the mitogen-activated protein kinase pathway in 3T3-L1 preadipocytes. Life Sci 86:791–797

    Article  CAS  PubMed  Google Scholar 

  • Kumaran T, Michaelbabu M, Selvaraj T, Albindhas S, Citarasu T (2010) Production of Anti WSSV IgY edible antibody using herbal immunoadjuvant Asparagus racemosus and its immunological influence against WSSV infection in Penaeus monodon. J Aquac Feed Sci Nutr 2(1):1–5

    Article  Google Scholar 

  • Lee JB, Hayashi K, Hayashi T, Sankawa U, Maeda M (1999) Antiviral activities against HSV-1, HCMV, and HIV-1 of rhamnan sulfate from Monostroma latissimum. Planta Med 65:439–441

    Article  PubMed  Google Scholar 

  • Leiro JM, Castro R, Arranz JA, Lamas J (2007) Immunomodulating activities of acidic sulphated polysaccharides obtained from the seaweed Ulva rigida C. Agardh. Int Immunopharmacol 7:879–888

    Article  CAS  PubMed  Google Scholar 

  • Li B, Lu F, Wei X, Zhao R (2008) Fucoidan: structure and bioactivity. Molecules 13:1671–1695

    Article  CAS  PubMed  Google Scholar 

  • Lowry OH, Rosebroogh NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  • Madhumathi M, Rengasamy R (2011) Antioxidant status of Penaeus monodon fed with Dunaliella salina supplemented diet and resistance against WSSV. Int J Eng Sci Technol 3(10):7249–7259

    Google Scholar 

  • Maeda M, Itami T, Kondo M, Henning O, Takahashi Y, Hirono I, Aoki T (1997) Characteristics of penaeied rod shaped DNA virus kuruma shrimp. In: Proceedings of NRIA International Workshop: new approaches to viral disease of aquatic animals, Jan. 21–24 National Research Institute of Aquaculture, Nansei, 218–228

  • Manilal A, Sujith S, Selvin J, Kiran GS, Shakir C (2009) In vivo antiviral activity of polysaccharide from the Indian green alga, Acrosiphonia orientalis (J. Agardh): potential implication in shrimp disease management world. J Fish Mar Sci 1(4):278–282

    CAS  Google Scholar 

  • Matou S, Helley D, Chabut D, Bros A, Fischer AM (2002) Effect of fucoidan on fibroblast growth factor-2-induced angiogenesis in vitro. Thromb Res 106:213–221

    Article  CAS  PubMed  Google Scholar 

  • Mautner HG, Gardner GM, Pratt R (1953) Antibiotic activity of seaweed extracts. II. Rhodomela larix. J Am Pharm Assoc Sci 42:294–296

    Article  CAS  Google Scholar 

  • Menasveta P (2002) Improved shrimp grow out systems for disease prevention and environmental sustainability in Asia. Rev Fish Sci 10:391–402

    Article  Google Scholar 

  • Namita R, Kumar S, Jaganmohan S, Murugan V (2007) DNA vaccines encoding viral envelope proteins confer protective immunity against WSSV in black tiger shrimp. Vaccine 25:2778–2786

    Article  Google Scholar 

  • Ogino C, Taki C (1957) Studies on the tannins of brown algae Sargassium ringgoldianum. Harv J Tokyo Univ Fish 43:1–5

    CAS  Google Scholar 

  • Pascual C, Sánchez A, Vargas-Albores F, LeMoullac G, Rosas C (2003) Haemolymph metabolic variables and immune response in Litopenaeus setiferus adult males: effect of an extreme temperature. Aquaculture 218:637–650

    Article  CAS  Google Scholar 

  • Pengzhan Y, Quanbin Z, Ning L, Zuhong X, Yanmei W, Zhi’en LJ (2003) Polysaccharides from Ulva pertusa and preliminary studies on their antihyperlipidemia activity. J Appl Phycol 15:21–27

  • Peters RI, Long GE (1973) Some alternation in haemolymph protein of Hemiorapaus nudus associated with temperature acculimation. Comp Biochem Physiol 46A:207–213

    Article  Google Scholar 

  • Qi H, Zhang Q, Zhao T, Hu R, Zhang K, Li Z (2009) In vitro antioxidant activity of acetylated and benzoylated derivatives of polysaccharide extracted from Ulva pertusa (Chlorophyta). Bioorg Med Chem Lett 16:2441–2445

  • Rameshthangam P, Ramasamy P (2007) Antiviral activity of bis(2-methylheptyl)phthalate isolated from Pongamia pinnata leaves against white spot syndrome virus of penaeus monodon fabricius. Virus Res 126:38–44

    Article  CAS  PubMed  Google Scholar 

  • Ratcliffe NA, Rowley AF (1979) Role of hemocytes in defense against biological agents. In: Gupta AP (ed) Insect hemocytes-development forms, functions and techniques. Cambridge University Press, Cambridge, pp 331–414

    Chapter  Google Scholar 

  • Ravi AV, Musthafa KS, Jegathammbal G, Kathiresan K, Pandian SK (2007) Screening and evaluation of probiotics as a bio control agent against pathogenic Vibrios in marine aquaculture. Lett Appl Microbiol 45:219–223

    Article  CAS  PubMed  Google Scholar 

  • Roe JR (1955) The determination of sugar in blood and spinal fluid with anthrone reagent. J Biol Chem 20:335–343

    Google Scholar 

  • Sahul Hameed AS (1989) Studies on the pathobiology of penaeid larvae and post larvae. PhD thesis, Cochin University of Science and Technology, Cochin, India, 243

  • Siamopoulou P, Bimplakis A, Iliopoulou D, Vagias C, Cos P, Berghe DV, Roussis V (2004) Diterpenes from the brown algae Dictyota dichotoma and Dictyota linearis. Phytochemistry 65:2025–2030

    Article  CAS  PubMed  Google Scholar 

  • Siddhanta AK, Ramavat BK, Chauhan VD, Achari B, Dutta PK, Pakrashi SC (1991) Sulphoglycolipid from the green algae Enteromorpha flexuosa (Wulf). J Agric Bot Mar 34:365–367

  • Siddhanta AK, Mody KH, Ramavat BK, Chauhan VD, Charma M, Garg HS (1995) Characterisation of Sulphoglycolipid from the red algae Laurencia pedicularioides. Bot Mar 38:329–331

    Article  CAS  Google Scholar 

  • Sieburth J, Conover JT (1965) Sargassun tannin an antibiotic which retards fouling. Nature 208:52–53

    Article  Google Scholar 

  • Small HJ, Pagenkopp KM (2011) Reservoirs and alternate hosts for pathogens of commercially important crustaceans: a review. J Invertebr Pathol 106:153–164

    Article  PubMed  Google Scholar 

  • Syed Musthaq S, Madhan S, Sahul Hameed AS, Kwang J (2009) Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon. Virology 391:315–324

    Article  CAS  PubMed  Google Scholar 

  • Takahashi Y, Itami T, Maeda M, Suzuki N, Kasornchandra J, Supamattaya K (1996) Polymerase chain reaction (PCR) amplification of bacilliform virus (RV-P) DNA in Penaeus japonicus Bate and systemic ectodermal and mesodermal baculovirus (SEMBV) DNA in Penaeus monodon Fabricices. J Fish Dis 19:399–403

    Article  CAS  Google Scholar 

  • Takahashi H, Ichimiya S, Nimura Y, Watanabe M, Furusato M, Wakui S, Yatani R, Aizawa S, Nakagawara A (1998) Mutation, allelotyping, and transcription analyses of the p73 gene in prostatic carcinoma. Cancer Res 58:2076–2077

    CAS  PubMed  Google Scholar 

  • Takahashi Y, Kondo M, Itami T, Honda T, Inagawa H, Niahizawa T (2000) Enhancement of disease resistance against penaeid acute viraemia and induction of virus-inactivating activity in haemolymph of kuruma shrimp, Penaeus japonicus by oral administration of Pantoea agglomerans lipopolysaccharide (LPS). Fish Shellfish Immunol 10:555–558

    Article  CAS  PubMed  Google Scholar 

  • Velmurugan S, Raman K, Thangaviji V, Donio MBS, Jenifer JA, Citarasu T (2012) Screening, and characterization of antimicrobial secondary metabolites from Halomonas salifodinae MPMTC and it’s in vivo antiviral influence on Indian White shrimp Fenneropenaeus indicus against WSSV infection. J King Saud Univ–Sci 25:181–190

  • Wijesekara I, Pangestuti R, Kim SK (2011) Biological activities and potential health benefits of sulfated polysaccharides derived from marine algae. Carbohydr Polym 84:1

    Article  Google Scholar 

  • Wongprasert K, Rudtanatip T, Praiboon J (2013) Immunostimulatory activity of sulfated galactans isolated from the red seaweed Gracilaria fisheri and development of resistance against white spot syndrome virus (WSSV) in shrimp. Fish Shellfish Immunol (in press)

  • Yeh ST, Lin YC, Huang CL, Chen JC (2010) White shrimp Litopenaeus vannamei that received the hot-water extract of Gracilaria tenuistipitata showed protective innate immunity and up-regulation of gene expressions after low-salinity stress. Fish Shellfish Immunol 28(5–6):887–894

    Article  CAS  PubMed  Google Scholar 

  • Yoganandhan K, Thirupathi S, Hameed ASS (2003) Biochemical, physiological and hematological changes in white spot syndrome virus-infected shrimp Penaeus indicus. Aquaculture 221:1–11

    Article  CAS  Google Scholar 

  • Yogeeswaran A, Velmurugan S, Punitha SMJ, Babu MM, Selvaraj T, Kumaran T, Citarasu T (2012) Protection of Penaeus monodon against White Spot Syndrome Virus by inactivated vaccine with herbal immunostimulants. Fish Shellfish Immunol 32:1058–1067

    Article  CAS  PubMed  Google Scholar 

  • Zhang HJ, Mao WJ, Fang F, Li HY, Sun HH, Chen Y, Qi XH (2008) Chemical characteristics and anti-coagulant activities of a sulfated polysaccharide and its fragments from Monostroma latissimun. Carbohydr Polym 71:428–434

Download references

Acknowledgments

The authors S. Velmurugan and Dr. T. Citarasu gratefully acknowledge the University Grants Commission (UGC), New Delhi, Government of India, for financial support in the form of a research grant [UGC-no. 37/271 (SR) dated 01.02.2010].

Author information

Authors and Affiliations

  1. Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam, 629 502, Tamilnadu, India

    S. Velmurugan, N. Jerin, M. Michael Babu, F. Bindhu, S. Albin Dhas & T. Citarasu

Authors
  1. S. Velmurugan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. Jerin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Michael Babu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F. Bindhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S. Albin Dhas
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T. Citarasu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T. Citarasu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Velmurugan, S., Jerin, N., Michael Babu, M. et al. Screening and characterization of antiviral compounds from Enteromorpha flexuosa against white spot syndrome virus (WSSV) and its in vivo influence on Indian white shrimp Fenneropenaeus indicus . Aquacult Int 23, 65–80 (2015). https://doi.org/10.1007/s10499-014-9798-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10499-014-9798-y

Keywords

Search

Navigation