Examination of the hemolymph indices and innate immunity of the western white shrimp (Litopenaeus vannamei) fed with a combined extract of brown macroalgae and single-cell probiotic (Bacillus subtilis IS0)

Document Type : Research Paper

Authors

1 Professor in Department of Fisheries, Faculty of Marine Sciences, Chabahar Maritime University, Chabahar, Iran

2 Ph.D. in Aquatic Health, Offshore Fisheries Research Center, Iranian Fisheries Science Research Institute (IFSRI), Agricultural Research, Education and Extension Organization (AREEO), Chabahar, Iran

10.22124/japb.2025.28719.1555

Abstract

This study was conducted to evaluate the effect of the aqueous extract combination of brown macroalgae and the bacterium Bacillus subtilis IS0 on the hemolymph indices and non-specific immunity of the white shrimp (Litopenaeus vannamei). A total of 2400 post-larvae with an average weight of 29±0.87mg were stocked at a density of 100 individuals in 8 groups (with three replicates). The control group received a diet without the combined macroalgae extract (MPE) and single cell probiotic (P), while the other groups were fed diets containing 15g/kg MPE, 1% (P1), 2% (P2) and 3% (P3) probiotics and combinations of the macroalgae extract with the probiotic (P1+MPE, P2+MPE and P3+MPE) for a period of 60 days. The results showed that the highest total hemocyte count was observed in the groups fed with P2+MPE and P3+MPE, at 15.36±0.35 and 15.10±0.36 ×106/mL), respectively, which were significantly different from the other experimental groups and control (P<0.05). The simultaneous use of an aqueous extract of macroalgae with different concentrations of single-cell probiotics significantly increased the number of granular, semi-granular and hyaline cells compared to the control group and other experimental groups. The highest counts of these cells were found in the groups fed with P2+MPE and P3+MPE, showing a significant difference from the other experimental groups (P<0.05). The highest protein level (111.47±9.21mg/mL) and the lowest glucose level (9.0±0.91mg/dL) were observed in the group fed P2+MPE, showing a significant difference compared to the control and other experimental groups (P<0.05). The results of this study demonstrated that the simultaneous use of the combined extract of brown macroalgae and 2% single-cell probiotic improved hemolymph indices and innate immune system in white shrimp.

Keywords

Main Subjects

References

Abdollahi Arpanahi D., Jafaryan H., Soltani M., Naderi Samani M. and Hasanpour Fatahi A. 2019. Comparison of commercial and indigenous Bacillus (Bacillus subtilis, Bacillus licheniformis) effects on some immune responses and serum enzymes activity in white leg shrimp post-larvae (Litopenaeus vannamei). Journal of Veterinary Research, 74(1): 84–92. doi: 10.22059/jvr.2017.221169.2543
Adel M., El-Sandy A.F.M., Yeganeh S., Dadar M. and Giri S.S. 2017. Effect of potential probiotic Lactococcus lactis subsp. lactis on growth performance, intestinal microbiota, digestive enzyme activities, and disease resistance of Litopenaeus vannamei. Probiotics and Antimicrobial Proteins, 9: 150–156. doi: 10.1007/s12602-016-9235-9
Aguirre-Guzman G., Sanchez-Martinez J.G., Campa-Cordova A.I., Luna-Gonzalez A. and Ascencio F. 2009. Penaeid shrimp immune system. The Thai Journal of Veterinary Medicine, 39(3): 205–215. doi: 10.56808/2985-1130.2 175
Akbary P. and Aminikhoei Z. 2018. Effect of polysaccharides extracts of algae Ulva rigida on growth, antioxidant, immune response and resistance of shrimp, Litopenaeus vannamei against Photobacterium damselae. Aquaculture Research, 49(7): 2503–2510. doi: 10.1111/are. 13710
Akbary P., Adeshina I. and Jahanbakhshi A. 2020. Growth performance, digestive enzymes, antioxidant activity and immune responses of Litopenaeus vannamei fed with Jania adhaerens J.V. supplemented diet against Photobacterium damselae infection. Animal Feed Science and Technology, 270: 114696–11473. doi: 10.1016/j.anifeedsci.20 20.114696
Barton B.A. 2002. Stress in fishes: A diversity of responses with particular reference to changes in circulating corticosteroids. Integrative and Comparative Biology, 42(3): 517–525. doi: 10. 1093/icb/42.3.517
Butt U.D., Lin N., Akhter N., Siddiqui T., Li S. and Wu B. 2021. Overview of the latest developments in the role of probiotics, prebiotics and syn-biotics in shrimp aquaculture. Fish and Shellfish Immunology 114: 263–281. doi: 10.1016/j.fsi.2021.05. 003
Castillo S., Rosales M., Pohlenz C. and Gatlin D.M. 2014. Effects              of organic acids on growth performance and digestive enzyme activities of juvenile red drum Sciaenops ocellatus. Aquaculture, 433: 6–12. doi: 10.1016/j.aquacultu re.2014.05.038
Chen Y.Y., Chen J.C., Lin Y.C., Putra D.F., Kitikiew S., Li C.C., Hsieh J.F., Liou C.H. and Yeh S.T. 2014. Shrimp that have received carrageenan via immersion and diet exhibit immunocompetence in phago-cytosis despite a post-plateau in immune parameters. Fish and Shellfish Immunology, 36(2): 352–366. doi: 10.1016/j.fsi.2013.12.004
Cheng Y. 2019. The growth performance and nonspecific immunity of red swamp crayfish Procambarus clarkia affected                    by dietary Rhodiola rosea polysaccharide. Fish and Shellfish Immunology, 93: 796–800. doi: 10.1016/j.fsi.2019.08.046
Choi Y.H., Kim K.W., Han H.S., Nam T.J. and Lee B.J. 2014. Dietary Hizikia fusiformis glycolprotein-induced IGF I and IGF-BP3 associated somatic growth, polyunsaturated fatty acid metabolism and immunity in juvenile olive flounder Paralichthys olivaceus. Comparative Biochemistry and Physiology (A), 167: 1–6. doi: 10.1016/j.cbpa.2013.09.011
Choi Y.H., Lee B.J. and Nam T.J. 2015. Effect of dietary inclusion of Pyropia yezoensis extract on biochemical and immune responses of olive flounder Paralichthys olivaceus. Aquaculture, 435: 347–353. doi: 10.1016/j.aquaculture.2014.10.010
Chojnacka K.W., Agnieszka S., Zuzanna W. and Lukazy T. 2012. Biologically active compounds in seaweed extracts- The prospects for the application. The Open Conference Proceedings Journal, 3(22): 20–28.
Chouhury S., Sree A., Mukherjee S.C., Pattnik P. and Bapuji M. 2005. In vitro antibacterial activity of extracts of selected marine algae and mangroves against fish pathogens. Asian Fish Science, 18: 85–94. doi: 10.3390/app12031148
Decamp O., Moriarty D.J.W. and Lavens P. 2008. Probiotics for shrimp larviculture: Review of field data from Asia and Latin America. Aquaculture Research, 39: 334–338. doi: 10.1111/j.1365-2109.2007.01664.x
Eidi Ghaleghazi F., Noori A. and Hosseinifar S.H. 2021. The effects of dietary supplementation of Sargassum ilicifolium hot-water extract on some haemolymph physiological parameters of white leg shrimp, Penaeus vannamei. Aquaculture Sciences, 9(1): 1–14.
Ellis A.E. 1990. Lysozyme assays. P: 101–103. In: Stolen J.S., Fletcher T.C., Anderson D.P., Roberson B.S. and Van Muiswinkel W.B. (Eds.). Techniques in Fish Immunology. SOS Publications, USA.
Farzanfar A. 2006. The use of probiotics in shrimp aquaculture. FEMS Immunology and Medical Microbiology, 48: 149–15.
Ghaednia B., Mehrabi M.R., Mirbakhsh M., Yeganeh V., Hoseinkhezri P., Garibi G. and Ghaffar Jabbari A. 2011. Effect of hot water extract of brown seaweed Sargassum glaucescens via immersion route on immune responses of Fenneropenaeus indicus. Iranian Journal of Fisheries Sciences, 10(4): 616–630. doi: 10.22092/IJFS.2018.114168
Ghaednia B., Mirbakhsh M. and Zorrieh Zahra M.J. 2020. Effect of Bacillus subtilis IS02 in food on health, immunity indexes and prevention against white spot disease in Litopenaeus vannamei. Journal of Aquaculture Development, 14(3) :87–99.
Jiang G., Yu R. and Zhou M. 2004. Modulatory effects of ammonia -N on the immune system of Penaeus japonicus to virulence of white spot syndrome virus. Aquaculture, 241: 61–75. doi: 10.1016/j.aquacultu re.2007.12.019
Jiravanichpaisal P., Lee B.L. and Soderhall K. 2006. Cell-mediated immunity in arthropods: Hemato-poiesis, coagulation, melanization and opsonization. Immunobiology, 211: 213–236. doi: 10.1016/j.imbio. 2005.10.015
Kazemi M., Abediankenari A. and Rabiei R. 2018. Effect of marine macroalgae on growth performance and immune response in rainbow trout fingerlings. Journal of Fisheries Science and Technology, 7(1): 9–16.
Kewcharoen W. and Srisapoome P. 2019. Probiotic effects of Bacillus spp. from Pacific white shrimp (Litopenaeus vannamei) on water quality and shrimp growth, immune responses, and resistance to Vibrio parahaemolyticus (AHPND strains). Fish and Shellfish Immunology, 94: 175–189. doi: 10.1016/j.fsi.2019.09.013
Lim S.Y., Loo K.W. and Wong W.L. 2019. Synergistic antimicrobial effect of a seaweed-probiotic blend against acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio parahaemolyticus. Probiotics and Antimicrobial Proteins, 12(3): 906–917. doi: 10.1007/s12602-019-09616-8
Liu K.F., Chiu C.H., Shiu Y.L., Cheng W. and Liu C.H. 2010. Effects of the probiotic, Bacillus subtilis E20, on the survival, development, stress tolerance, and immune status of white shrimp, Litopenaeus vannamei larvae. Fish and Shellfish Immunology, 28(5-6): 837–844. doi: 10.1016/j.fsi.2010. 01.012
Liu S., Jiang X., Hu X., Gong J., Hwang H. and Mai K. 2004. Effects of temperature on non-specific immune parameters in two scallop species: Argopecten irradians (Lamarck 1819) and Chlamys farreri (Jones & Preston 1904). Aquaculture Research, 35: 678–682. doi: 10.1111/j.1365-2109. 2004.01065.x
Merrifield D.L., Dimitroglou A., Foey A., Davies S.J., Baker R.T.M., Bogwald J., Castex M. and Ringo E. 2010. The current status and future focus of probiotic and prebiotic applications for salmonids. Aquaculture, 302: 1–18. doi: 10.1016/j.aquaculture.2010. 02.007
Meshkini S. and Tafi A.A. 2016. Presentation and usage of immunostimulators in aquaculture. Agricultural and Natural Resources Engineering, 46(12): 50–55. doi: 10.1016/j.fsi.2005.03.008
Monier M.N., Kabary H., Elfeky A., Saadony S., El-Hamed N.N.B., Abd Eissa M.E.H. and Eissa E.S.H. 2023. The effects of Bacillus species probiotics (Bacillus subtilis and B. licheniformis) on the water quality, immune responses, and resistance of white leg shrimp (Litopenaeus vannamei) against Fusarium solani infection. Aquaculture International, 31: 3437–3455. doi: 10.1007/s10499-023-01136-1
Morshedi V., Nafisi Bahabadi M., Azodi M., Modaresi M. and Cheraghi S. 2015. Effects of dietary probiotic (Lacrobacillus plantarum) on body composition, serum biochemical parameters and liver enzymes of Asian sea bass (Lates calcarifer). Journal of Marine Science and Technology, 14(2): 1–14. doi: 10.22113/jmst.20 15.8579
Munaei W., Yuhana M. and Widanarnin W. 2014. Effect of micro-encapsulated synbiotic at different frequencies for luminous vibriosis control in white shrimp (Litopenaeus vannamei). Microbiology Indonesia, 8(2): 73–80.
Pascual C., Sanchez A., Zenteno E., Cuzon G., Gabriela G., Brito R., Gelabert R., Hidalgo E. and Rosas C. 2006. Biochemical, physiological, and immunological changes during starvation in juveniles of Litopenaeus vannamei. Aquaculture, 251(2-4): 416–429. doi: 10.1016/j.aquaculture.2005.06.001
Perazzolo L.M., Gargioni R., Ogliari P. and Barracco M.A.A. 2002. Evaluation of some hemato-immunological parameters in the shrimp Farfantepenaeus paulensis submitted to environmental and physiological stress. Aquaculture, 214: 19–33. doi: 10.1016/S0044-84 86(02)00137-0
Pereira L. 2012. A review of the nutrient composition of selected edible seaweeds. P: 15–47. In: Pomin V.H. (Ed.). Seaweed: Ecology, Nutrient Composition and Medicinal Uses. Nova Science, USA.
Qiu Z., Xu Q., Li S., Zheng D., Zhang R., Zhao J. and Wang T. 2023. Effects of probiotics on the water quality, growth performance, immunity, digestion, and intestinal flora of giant freshwater prawn (Macrobrachium rosenbergii) in the biofloc culture system. Water, 15: 1211–1220. doi: 10.3390/w1506 1211
Rabiei R., Asadi M., Sohrabipour J., Nejadsattari T. and Majd A. 2007. Algae species diversity of Gracilaria salicornia on the Persian Gulf Coast‐Qeshm Island. Journal of Research Construction, 20(2): 43–57.
Rengpipat S., Phianphak W., Piyatiratitivorakul S. and Menasaveta P. 1998. Effects of a probiotic bacterium in black tiger shrimp Penaeus monodon survival and growth. Aquaculture, 167: 301–313. doi: 10.1016/S0044-8486 (98)00305-6
Salehpour R., Amrollahi Biuki N., Mohammadi M., Dashtiannasab A., Ebrahimnejad P. and Ghasemi A. 2022. Evaluation of growth performance and biochemical parameters of western white leg shrimp, Litopenaeus vannamei (Boone, 1931) fed with fucoidan enriched diet. Journal of Marine Science and Technology, 21(4): 41–54. doi: 10.22113/jmst.20 21.268814.2412
Sandeepa M.G. and Ammani K. 2015. Effect of probiotic bacterium on growth and biochemical parameters of shrimp Litopenaeus vannamei. International Journal of Recent Scientific Research, 6(2): 2871–2875. doi: 10.1016/j.aqrep.20 23.101873
Song L., Yu I., Lien W. and Huang C. 2003. Haemolymph parameters of Litopenaeus vannamei infected with Taura syndrome virus. Fish and Shellfish Immunology, 14: 317–331. doi: 10.1006/fsim.2002.04 40
Su C., Fan D., Pan L., Lu Y., Wang Y. and Zhang M. 2020. Effects of Yu-Ping-Feng polysaccharides (YPS) on the immune response, intestinal microbiota, disease resistance and growth performance of Litopenaeus vannamei. Fish and Shellfish Immunology, 105: 104–116. doi: 10.1016/j.fsi.2020.07.003
Sudaryono A., Chilmawati D. and Susilowati T. 2018. Oral administration of hot-water extract of tropical brown seaweed, Sargassum cristaefolium, to enhance immune response, stress tolerance, and resistance of white shrimp, Litopenaeus vannamei, to Vibrio parahaemolyticus. Journal of the World Aquaculture Society, 49: 877–888. doi: 10.1111/jwas.125 27
Tacon A., Cody J., Cnquest L., Divakaran S., Forster I. and Decamp O. 2002. Effect of culture system on the nutrition and growth performance of Pacific white shrimp Litopenaeus vannamei (Boone) fed different diets. Aquaculture Nutrition, 8: 121–137. doi: 10.1046/j.1365-2095.2002.0019 9.x
Tayag C.M., Lin Y.C., Li C.C., Liou C.H. and Chen J.C. 2010. Administration of the hot-water extract of Spirulina platensis enhanced the immune response of white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus. Fish and Shellfish Immunology, 28: 764–773. doi: 10.1016/j.fsi.2010.01.023
Trinder P. 1969. Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor. Annuals of Clinical Biochemistry, 6: 24–27. doi: 10.117 7/000456326900600108
Vaezi M., Esmaeili Feridooni A., Manaffar R. and Amini K. 2018. Effects of probiotic (Pediococcus acidilactici) on heamatological parameters, immunological responses and digestive enzymes of Astacus leptodactylus juveniles. Aquatic Physiology and Biotechnology, 6(3): 35–60. doi: 10.22124/japb.2018.8702.1197
Vatsos I.N. and Rebours C. 2015. Seaweed extracts as antimicrobial agents in aquaculture. Journal of Applied Phycology, 27(5): 2017–2035. doi:10.1007/s10811-014-0506-0
Wache Y., Auffray F., Gatesoupe F.J., Zambonino J., Gayet V. and Labbe L. 2006. Cross effects of the strain of dietary Saccharomyces cerevisiae and rearing conditions on the onset of intestinal microbiota and digestive enzymes in rainbow trout, Oncorhynchus mykiss, fry. Aquaculture, 258: 470–478. doi: 10.1016/j.aquaculture. 2006.04.002
Xue M., Wen C., Liang H., Ding M., Wu Y. and Li X. 2016. In vivo evaluation of the effects of commercial Bacillus probiotics on survival and development of Litopenaeus vannamei larvae during the early hatchery period. Aquaculture Research, 47: 1661–1669. doi: 10.1111/are.12719
Xue Q. and Renault T. 2000. Enzymatic activities in European flat oyster, Ostrea edulis, and Pacific oyster, Crassostrea gigas, hemolymph. Journal of Invertebrate Pathology, 76: 155–163. doi: 10.1006/jipa.2000.4965
Yang C., Chen N., Lu L., Chen S. and Lai C. 2014. Effect of mushroom beta glucan on immune and haemocyte response in shrimp Litopenaeus vannamei. Journal of Aquaculture Research Development, 5(6): 1–5 (1000275). doi: 10.3390/antiox11112282
Yeh S.T., Lin Y.C., Huang C.L. and Chen J.C. 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 and Shellfish Immunology, 28: 887–894. doi: 10.1016/j.fsi.2010.02.005
Yu M.C., Li Z.J., Lin H.Z., Wen G.L. and Ma S. 2008. Effects of dietary Bacillus and medicinal herbs on the growth, digestive enzyme activity, and serum biochemical parameters of the shrimp Litopenaeus vannamei. Aquaculture International, 16: 471–480. doi: 10.1007/s10499-007-9 159-1
Zhang Z., Shi X., Wu Z., Wu W., Zhao Q. and Li E. 2023. Macroalgae improve the growth and physiological health of white shrimp (Litopenaeus vannamei). Aquaculture Nutrition, 2023: 1–8 (8829291). doi: 10.1155/2023/88292 91
Zokaeifar H., Balcazar J.L., Saad C.R., Kamarudin M.S., Sijam K., Arshad A. and Nejat N. 2012. Effects of Bacillus subtilis on the growth performance, digestive enzymes, immune gene expression and disease resistance of white shrimp Litopenaeus vannamei. Fish and Shellfish Immunology, 33(4): 683–689. doi: 10.1016/j.fsi.20 12.05.027
Aquatic Physiology and Biotechnology
Volume 13, Issue 2
November 2025
Pages 137-160

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