Immunostimulant effect of nucleotide supplementation on shrimp Penaeus vannamei submitted to bacterial challenge with Vibrio parahaemolyticus
Abstract
Aim of the study: Vibriosis and other bacterial infections significantly challenge Pacific white shrimp (Penaeus vannamei) production. This study evaluated the immunostimulant effect of nucleotide supplementation on hemato-immunological parameters, digestive gland morphology, presumptive Vibrio count, and survival of Penaeus vannamei exposed to Vibrio parahaemolyticus.
Area of study: Aquaculture nutrition and health management, specifically focusing on the mitigation of bacterial disease (Vibriosis) in cultured Pacific white shrimp.
Material and methods: Three nucleotide supplementation levels were evaluated: NT75 (75 mg/kg), NT150 (150 mg/kg), and NT300 (300 mg/kg), alongside a control group (C). After 60 days of feeding the experimental diets, the shrimp were challenged by immersion in a bacterial suspension of Vibrio parahaemolyticus (106 CFU mL-1) for 30 days. Mortality and clinical signs were monitored, and hemato-immunological, histological, and bacteriological parameters were analyzed.
Main results: Nucleotide-supplemented treatments exhibited improved growth performance and reduced pre-challenge mortality. Following the bacterial challenge, all treatments showed an acute, albeit non-significant, change in total and differential hemocyte counts. However, histological analysis of the digestive gland revealed severe lesions, including tubule deformities and necrosis, across all treatment groups. On days 5 and 15, significant differences in Vibrio sp. counts within the digestive gland were observed. Specifically, differences were noted between Control (C) and NT75, and between NT75 and NT300, indicating varying bacterial colonization over time.
Research highlights: Higher nucleotide supplementation levels improved shrimp performance, reduced pre-challenge mortality, and enhanced the acute immune response following a bacterial challenge, suggesting their utility as an effective immunostimulant against Vibriosis.
Downloads
References
Abdel-Rahim M, Bahattab O, Nossir F, Al-Awthan Y, Khalil RH, Mohamed R, 2021. Dietary supplementation of brown seaweed and/or nucleotides improved shrimp performance, health status and cold-tolerant gene expression of juvenile whiteleg shrimp during the winter season. Mar Drugs 19(3): 175. https://doi.org/10.3390/md19030175
Agboola JO, Overland M, Skrede A, Hansen JO, 2021. Yeast as major protein-rich ingredient in aquafeeds: a review of the implications for aquaculture production. Rev Aquac 13(2): 949–970. https://doi.org/10.1111/raq.12507
APHA, 2012. Standard methods for the examination of water and wastewater. 22nd ed. American Public Health Association; American Water Works Association; Water Environment Federation, Washington, DC.
Avnimelech Y, 2012. Biofloc Technology. A Practical Guide Book. World Aquaculture Society, Louisiana (US).
Baumann P, Baumann L, Bang SS, Reichelt JL, 1980. Reevaluation of the taxonomy of Vibrio, Beneckea, and Photobacterium: Abolition of the genus Beneckea. Curr Microbiol 4: 127–132. https://doi.org/10.1007/BF02602814
Beçak W, Paulete J, 1976. Cytology and histology techniques. Scientific and Technical Books Editora, Rio de Janeiro.
Boone L, 1931. A collection of anomuran and macruran Crustacea from the Bay of Panama and the fresh waters of the Canal Zone. Bull Am Mus Nat Hist 63: 137–189.
Caro LFA, Mai HN, Noble B, Dhar AK, 2020. Acute hepatopancreatic necrosis disease (VPAHPND), a chronic disease in shrimp (Penaeus vannamei) population raised in Latin America. J Invertebr Pathol 174: 107424. https://doi.org/10.1016/j.jip.2020.107424
Celi M, Filiciotto F, Parrinello D, Buscaino G, Damiano MA, Cuttitta A, D’Angelo S, Mazzola S, Vazzana M, 2013. Physiological and agonistic behavioural response of Procambarus clarkii to an acoustic stimulus. J Exp Biol 216(4): 709–718. https://doi.org/10.1242/jeb.078865
de Souza Valente C, Wan AH, 2021. Vibrio and major commercially important vibriosis diseases in decapod crustaceans. J Invertebr Pathol 181: 107527. https://doi.org/10.1016/j.jip.2020.107527
Ding T, Song G, Liu X, Xu M, Li Y, 2021. Nucleotides as optimal candidates for essential nutrients in living organisms: A review. J Funct Foods 82: 104498. https://doi.org/10.1016/j.jff.2021.104498
FAO, 2024. The State of World Fisheries and Aquaculture. Blue transformation in action. FAO, Rome. https://doi.org/10.4060/cd0683en
Fernández JL, 2020. Estudio agroecológico del cultivo del maíz y sus potencialidades en la sustentabilidad de pequeñas fincas campesinas. Master’s thesis. Univ. Int. de Andalucía, Cádiz, Spain. 143 pp.
Fregoso-López MG, Morales-Covarrubias MS, Franco-Nava MA, Ramírez-Rochín J, Fierro-Sañudo JF, Ponce-Palafox JT, Páez-Osuna F, 2017. Histological alterations in gills of shrimp Litopenaeus vannamei in low-salinity waters under different stocking densities: Potential relationship with nitrogen compounds. Aquacult Res 48(12): 5854–5863. https://doi.org/10.1111/are.13408
Fries J, 1971. Análisis de trazas: métodos fotométricos comprobados. Darmstadt: Merck.
Fujino T, Miwatani T, Yasuda J, Kondo M, Takeda Y, Akita Y, Kotera K, Okada M, Nishimune H, Shimizu Y, Tamura T, Tamura Y, 1965. Taxonomic studies on the bacterial strains isolated from cases of “shirasu” food-poisoning (Pasteurella parahaemolytica) and related microorganisms. Biken J 8(2): 63–71.
Guertler C, Rieg T, Mejía-Ruíz CH, Lehmann M, Barracco MA, Perazzolo LM, 2013. Hemogram and survival rate of marine shrimp after silencing of WSSV by RNA interference. Pesqui Agropecu Bras 48: 983–990. https://doi.org/10.1590/S0100-204X2013000800025
He X, Abakari G, Tan H, Liu W, Luo G, 2023. Effects of different probiotics (Bacillus subtilis) addition strategies on a culture of Litopenaeus vannamei in biofloc technology (BFT) aquaculture system. Aquaculture 566: 739216. https://doi.org/10.1016/j.aquaculture.2022.739216
IBGE – Instituto Brasileiro de Geografia e Estatística, 2024. Produção da Pecuária Municipal 2023, v.51, 1–12. Produção da Pecuária Municipal.
Jarc E, Petan T, 2019. Focus: Organelles: Lipid droplets and the management of cellular stress. Yale J Biol Med 92(3): 435.
Javahery S, Noori A, Hoseinifar SH, 2019. Growth performance, immune response, and digestive enzyme activity in Pacific white shrimp, Penaeus vannamei Boone, 1931, fed dietary microbial lysozyme. Fish Shellfish Immunol 92: 528–535. https://doi.org/10.1016/j.fsi.2019.06.049
Johansson MW, Keyser P, Sritunyalucksana K, Söderhäll K, 2000. Crustacean haemocytes and haematopoiesis. Aquaculture 191(1–3): 45–52. https://doi.org/10.1016/S0044-8486(00)00418-X
Kobayashi T, 1963. A new selective isolation medium for the Vibrio group; on a modified Nakanishi’s medium (TCBS agar medium). Jpn J Bacteriol 18: 387–392.
Licona-Jain A, Racotta I, Angulo C, Luna-González A, Escamilla-Montes R, Cortés-Jacinto E, Morelos-Castro RM, Campa-Córdova ÁI, 2022. Combined administration routes of marine yeasts enhanced immune-related genes and protection of white shrimp (Penaeus vannamei) against Vibrio parahaemolyticus. Fish Shellfish Immunol 124: 192–200. https://doi.org/10.1016/j.fsi.2022.04.004
Lightner DV, 1996. A handbook of shrimp pathology and diagnostic procedures for diseases of cultured penaeid shrimp.
Lin X, Söderhäll I, 2011. Crustacean hematopoiesis and the astakine cytokines. Blood 117(24): 6417–6424. https://doi.org/10.1182/blood-2010-11-320614
Liu MJ, Liu S, Liu HP, 2021. Recent insights into hematopoiesis in crustaceans. Fish Shellfish Immunol Rep 2: 100040. https://doi.org/10.1016/j.fsirep.2021.100040
Lu J, Shi Y, Yao T, Bai C, Jiang J, Ye L, 2021. Gender differences in hemocyte immune parameters of Hong Kong oyster Crassostrea hongkongensis during immune stress. Front Immunol 12: 659469. https://doi.org/10.3389/fimmu.2021.659469
Mahdy MA, Jamal MT, Al-Harb M, Al-Mur BA, Haque MF, 2022. Use of yeasts in aquaculture nutrition and immunostimulation: A review. J Appl Biol Biotechnol 10(5): 59–65. https://doi.org/10.7324/JABB.2022.100507
Mok WY, Yong ASK, Mohamad Lal MT, Shapawi R, Kim YS, 2021. Dietary guanosine-monophosphate improves growth performance, feed utilization and intestinal morphology of whiteleg shrimp (Litopenaeus vannamei) maintained on soybean meal-based diets. Aquacult Res 52(4): 1453–1462. https://doi.org/10.1111/are.14999
Mulyadi IN, Iba W, 2020. Efficacy of seaweed (Sargassum sp.) extract to prevent vibriosis in white shrimp (Litopenaeus vannamei) juvenile. Int J Zool Res 16. https://doi.org/10.3923/ijzr.2020.1.11
Novriadi R, Ilham I, Roigé O, Segarra S, 2021. Effects of dietary nucleotides supplementation on growth, total haemocyte count, lysozyme activity and survival upon challenge with Vibrio harveyi in Pacific white shrimp, Litopenaeus vannamei. Aquacult Rep 21: 100840. https://doi.org/10.1016/j.aqrep.2021.100840
Poleksić V, Mitrović-Tutundžić V, 1994. Fish gills as a monitor of sublethal and chronic effects of pollution. In: Sublethal and chronic effects of pollutants on freshwater fish. pp. 339–352.
Rairat T, Chuchird N, Keetanon A, Carcano P, Comi M, Koppe W, 2022. Effects of dietary yeast-derived nucleotide and RNA on growth performance, survival, immune responses, and resistance to Vibrio parahaemolyticus infection in Pacific white shrimp (Litopenaeus vannamei). Aquacult Rep 27: 101352. https://doi.org/10.1016/j.aqrep.2022.101352
Segarra S, Chau T, Hoang P, Tran L, 2023. Immunoregulation and resistance to aquatic pathogens with dietary nucleotides in Pacific white shrimp, Litopenaeus vannamei. Fishes 8(6): 308. https://doi.org/10.1016/j.aqrep.2022.101352
Söderhäll I, 2016. Crustacean hematopoiesis. Dev Comp Immunol 58: 129–141. https://doi.org/10.1016/j.dci.2015.12.009
Suleman S, Andayani S, Yuniarti A, 2019. Potential of Ulva lactuta crude extract in increasing total haemocyte count (THC) and fagocytic activity in vaname shrimp (Litopenaeus vannamei). Samakia: J Ilmu Perikanan 10(1): 01–07. https://doi.org/10.35316/jsapi.v10i1.230
Tran L, Nunan L, Redman RM, Mohney LL, Pantoja CR, Fitzsimmons K, Lightner DV, 2013. Determination of the infectious nature of the agent of acute hepatopancreatic necrosis syndrome affecting penaeid shrimp. Dis Aquat Org 105(1): 45–55. https://doi.org/10.3354/dao02621
Van Wyk P, 1999. Nutrition and feeding of Litopenaeus vannamei in intensive culture systems. In: Farming marine shrimp in recirculating freshwater systems. pp. 220.
Van Wyk P, Scarpa J, 1999. Water quality requirements and management. In: Farming marine shrimp in recirculating freshwater systems. pp. 141–161.
Vijayaram S, Sun YZ, Zuorro A, Ghafarifarsani H, Van Doan H, Hoseinifar SH, 2022. Bioactive immunostimulants as health-promoting feed additives in aquaculture: A review. Fish Shellfish Immunol. https://doi.org/10.1016/j.fsi.2022.09.011
Vogt G, 2022. Cytology, function and dynamics of stem and progenitor cells in decapod crustaceans. Biol Rev 97(2): 817–850. https://doi.org/10.1111/brv.12824
World Organisation for Animal Health, 2022. Aquatic animal health code (25th ed., Section 9). World Organisation for Animal Health, Paris. https://www.woah.org/en/what-we-do/standards/codes-and-manuals/aquatic-code-online-access/
Yin X, Zhuang X, Liao M, Huang L, Cui Q, Liu C, Wang W, 2022. Transcriptome analysis of Pacific white shrimp (Litopenaeus vannamei) hepatopancreas challenged by Vibrio alginolyticus reveals lipid metabolic disturbance. Fish Shellfish Immunol 123: 238–247. https://doi.org/10.1016/j.fsi.2022.03.004
Yong ASK, Mok WY, Tamrin MLM, Shapawi R, Kim YS, 2020. Effects of dietary nucleotides on growth, survival and metabolic response in whiteleg shrimp, Litopenaeus vannamei, against ammonia stress condition. Aquacult Res 51(6): 2252–2260. https://doi.org/10.1111/are.14570
Zar JH, 2013. Biostatistical Analysis. J Am Stat Assoc.
Copyright (c) 2025 Consejo Superior de Investigaciones Científicas (CSIC)
This work is licensed under a Creative Commons Attribution 4.0 International License.
© CSIC. Manuscripts published in both the print and online versions of this journal are the property of the Consejo Superior de Investigaciones Científicas, and quoting this source is a requirement for any partial or full reproduction.
All contents of this electronic edition, except where otherwise noted, are distributed under a Creative Commons Attribution 4.0 International (CC BY 4.0) licence. You may read the basic information and the legal text of the licence. The indication of the CC BY 4.0 licence must be expressly stated in this way when necessary.
Self-archiving in repositories, personal webpages or similar, of any version other than the final version of the work produced by the publisher, is not allowed.
