Abstract
Vibrio scophthalmi and Aeromonas salmonicida can cause high turbot mortality and huge economic losses. Presently, vaccination is the most promising method for preventing communicable diseases. In this study, we used formalin to kill V. scophthalmi and A. salmonicida cells, and mixed with the mineralized oil adjuvant (Montanide™ ISA 763 AVG) to prepare the bivalent inactivated vaccine. The results showed that turbot inoculated with the bivalent inactivated vaccine exhibited strong tolerance to the infection of V. scophthalmi and A. salmonicida, and no obvious clinical symptoms and pathological changes were observed. The activities of enzymes lysozyme, acid phosphatase and complement C3 had significantly increased after the vaccination. The antibody titer response of vaccinated turbot was greatly boosted, which was positively connected with the immunological impact according to ELISA results. Simultaneously, the expression levels of immune-related genes such as MHC-IIα, MHC-IIβ, CD4, CD8, TNF-α and IL-1β were up-regulated, demonstrating that it might stimulate humoral and cellular immunological response in turbot. These findings highlight the potential of the bivalent inactivated vaccine for controlling V. scophthalmi and A. salmonicida infections in turbot.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Abu Nor, N., Zamri-Saad, M., Md Yasin, I. S., Salleh, A., Mustaffa-Kamal, F., Matori, M., F., et al., 2020. Efficacy of whole cell inactivated Vibrio harveyi vaccine against Vibriosis in a marine red hybrid tilapia (Oreochromis niloticus × O. mossambicus) model. Vaccines (Basel), 8(4): 734–748, DOI: https://doi.org/10.3390/vaccines8040734.
Bao, P., Sun, X., Liu, Q., Zhang, Y., and Liu, X., 2019. Synergistic effect of a combined live Vibrio anguillarum and Edwardsiella piscicida vaccine in turbot. Fish and Shellfish Immunology, 88: 84–90, DOI: https://doi.org/10.1016/j.fsi.2019.02.014.
Bui, C. T., Shollenberger, L. M., Paterson, Y., and Harn, D. A., 2014. Schistosoma mansoni soluble egg antigens enhance Listeria monocytogenes vector HIV-1 vaccine induction of cytotoxic T cells. Clinical and Vaccine Immunology, 21(9): 1232–1239, DOI: https://doi.org/10.1128/CVI.00138-14.
Caipang, C. M., Brinchmann, M. F., and Kiron, V., 2009. Profiling gene expression in the spleen of Atlantic cod, Gadus morhua upon vaccination with Vibrio anguillarum antigen. Comparative Biochemistry Physiology part B: Biochemistry and Molecular Biology, 153(3): 261–267, DOI: https://doi.org/10.1016/j.cbpb.2009.03.005.
Castro, N., Toranzo, A. E., Barja, J. L., Nunez, S., and Magarinos, B., 2006. Characterization of Edwardsiella tarda strains isolated from turbot, Psetta maxima (L.). Journal of Fish Diseases, 29(9): 541–547, DOI: https://doi.org/10.1111/j.1365-2761.2006.00750.x.
Cerda-Cuellar, M., Rossello-Mora, R. A., Lalucat, J., Jofre, J., and Blanch, A., 1997. Vibrio scophthalmi sp. nov., a new species from turbot (Scophthalmus maximus). International Journal of Systematic Bacteriology, 47(1): 58–61, DOI: https://doi.org/10.1099/00207713-47-1-58.
Chen, J., Liao, M. Y., Gao, X. L., Zhong, Q., and Cheng, X., 2013. IL-17A induces pro-inflammatory cytokines production in macrophages via MAPKinases, NF-κB and AP-1. Cellular Physiology Biochemistry, 32(5): 1265–1274, DOI: https://doi.org/10.1159/000354525.
Chen, Y., Cai, S., and Jian, J., 2019. Protection against Vibrio alginolyticus in pearl gentian grouper (♀Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatu) immunized with an acfA-deletion live attenuated vaccine. Fish and Shellfish Immunology, 86: 875–881, DOI: https://doi.org/10.1016/j.fsi.2018.12.030.
Dehghani, S., 2012. Efficacy of formalin-killed, heat-killed and lipopolysaccharide vaccines against motile aeromonads infection in rainbow trout (Oncorhynchus mykiss). Global Veterinaria, 9(4): 409–415, DOI: https://doi.org/10.5829/idosi.gv.2012.9.4.6591.
Farto, R., Milton, D. L., Bermudez, M. B., and Nieto, T. P., 2011. Colonization of turbot tissues by virulent and avirulent Aeromonas salmonicida subsp. salmonicida strains during infection. Diseases of Aquatic Organisms, 95(2): 167–173, DOI: https://doi.org/10.3354/dao02342.
Gudding, R., and Van Muiswinkel, B., 2013. A history of fish vaccination: Science-based disease prevention in aquaculture. Fish and Shellfish Immunology, 35(6): 1683–1688, DOI: https://doi.org/10.1016/j.fsi.2013.09.031.
Gwab, C., Scab, C., Ywab, C., Smab, C., and Yhab, C., 2020. Immune effect of Vibrio harveyi formalin-killed cells vaccine combined with chitosan oligosaccharide and astragalus polysaccharides in ♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatus. Fish and Shellfish Immunology, 98: 186–192, DOI: https://doi.org/10.1016/j.fsi.2020.01.015.
Jiang, N., Luo, L., Xing, W., Li, T., Yuan, D., Xu, G., et al., 2019. Generation and immunity effect evaluation of biotechnology-derived Aeromonas veronii ghost by PhiX174 gene E-mediated inactivation in koi (Cyprinus carprio Koi). Fish and Shellfish Immunology, 86: 327–334, DOI: https://doi.org/10.1016/j.fsi.2018.07.042.
Khezerlou, A., Alizadeh-Sani, M., Azizi-Lalabadi, M., and Ehsani, A., 2018. Nanoparticles and their antimicrobial properties against pathogens including bacteria, fungi, parasites and viruses. Microbial Pathogenesis, 123(20): 505–526, DOI: https://doi.org/10.1016/j.micpath.2018.08.008.
Li, K., Donaldson, B., Young, V., Ward, V., Jackson, C., Baird, M., et al., 2017. Adoptive cell therapy with CD4+ T helper 1 cells and CD8+ cytotoxic T cells enhances complete rejection of an established tumour, leading to generation of endogenous memory responses to non-targeted tumour epitopes. Clinical Translational Immunology, 6(10): 160, DOI: https://doi.org/10.1038/cti.2017.37.
Lim, J., and Hong, S., 2020. Characterization of Aeromonas salmonicida and A. sobria isolated from cultured salmonid fish in Korea and development of a vaccine against furunculosis. Journal of Fish Diseases, 43(5): 609–620, DOI: https://doi.org/10.1111/jfd.13158.
Liu, X., Zhang, H., Jiao, C., Liu, Q., Zhang, Y., and Xiao, J., 2017. Flagellin enhances the immunoprotection of formalin-inactivated Edwardsiella tarda vaccine in turbot. Vaccine, 35(2): 369–374, DOI: https://doi.org/10.1016/j.vaccine.2016.11.031.
Lopez-Vazquez, C., Dopazo, C. P., Barja, J. L., and Bandin, I., 2007. Experimental infection of turbot, Psetta maxima (L.), with strains of viral haemorrhagic septicaemia virus isolated from wild and farmed marine fish. Journal of Fish Diseases, 30(5): 303–312, DOI: https://doi.org/10.1111/j.1365-2761.2007.00815.x.
Marana, M. H., Jorgensen, L. V., Skov, J., Chettri, J. K., Holm Mattsson, A., Dalsgaard, I., et al., 2017. Subunit vaccine candidates against Aeromonas salmonicida in rainbow trout Oncorhynchus mykiss. PLoS One, 12(2): 17–19, DOI: https://doi.org/10.1371/journal.pone.0171944.
Marana, M. H., Skov, J., Chettri, J. K., Krossoy, B., Dalsgaard, I., Kania, P. W., et al., 2017. Positive correlation between Aeromonas salmonicida vaccine antigen concentration and protection in vaccinated rainbow trout Oncorhynchus mykiss evaluated by a tail fin infection model. Journal of Fish Diseases, 40(4): 507–516, DOI: https://doi.org/10.1111/jfd.12527.
Nascimento, D. S., Pereira, P. J., Reis, M. I., do Vale, A., Zou, J., Silva, M. T., et al., 2007. Molecular cloning and expression analysis of sea bass (Dicentrarchus labrax L.) tumor necrosis factor-α (TNF-α). Fish and Shellfish Immunology, 23(3): 701–710, DOI: https://doi.org/10.1016/j.fsi.2007.02.003.
Nguyen, H. T., Thu Nguyen, T. T., Tsai, M. A., Ya-Zhen, E., Wang, P. C., and Chen, S. C., 2017. A formalin-inactivated vaccine provides good protection against Vibrio harveyi infection in orange-spotted grouper (Epinephelus coioides). Fish and Shellfish Immunology, 65: 118–126, DOI: https://doi.org/10.1016/j.fsi.2017.04.008.
Pei, C., Gao, Y., Sun, X., Li, L., and Kong, X., 2019. A developed subunit vaccine based on fiber protein VP56 of grass carp reovirus providing immune protection against grass carp hemorrhagic disease. Fish and Shellfish Immunology, 90: 12–19, DOI: https://doi.org/10.1016/j.fsi.2019.04.055.
Pulpipat, T., Maekawa, S., Wang, P. C., and Chen, S. C., 2020. Immune responses and protective efficacy of a formalin-killed Francisella noatunensis subsp. orientalis vaccine evaluated through intraperitoneal and immersion challenge methods in Oreochromis niloticus. Vaccines (Basel), 8(2): 163–177, DOI: https://doi.org/10.3390/vaccines8020163.
Qiao, G., Lee, D. C., Woo, S. H., Li, H., Xu, D. H., and Park, S. I., 2012. Microbiological characteristics of Vibrio scophthalmi isolates from diseased olive flounder Paralichthys olivaceus. Fisheries Science, 78(4): 853–863, DOI: https://doi.org/10.1007/s12562-012-0502-8.
Ramos-Espinoza, F. C., Cueva-Quiroz, V. A., Yunis-Aguinaga, J., Alvarez-Rubio, N. C., Paganoti de Mello, N., and Engracia de Moraes, J. R., 2020. Efficacy of two adjuvants administrated with a novel hydrogen peroxide-inactivated vaccine against Streptococcus agalactiae in Nile tilapia fingerlings. Fish and Shellfish Immunology, 105: 350–358, DOI: https://doi.org/10.1016/j.fsi.2020.07.051.
Saurabh, S., and Sahoo., P. K., 2008. Lysozyme: An important defence molecule of fish innate immune system. Aquaculture Research, 39: 223–239, DOI: https://doi.org/10.1111/j.1365-2109.2007.01883.x.
Shona, K., and Fish, W. J., 2007. The innate immune response of finfish–A review of current knowledge. Fish and Shellfish Immunology, 23: 1127–111, DOI: https://doi.org/10.1016/j.fsi.2007.06.005.
Song, M. F., Kang, Y. H., Zhang, D. X., Chen, L., Bi, J. F., Zhang, H. P., et al., 2018. Immunogenicity of extracellular products from an inactivated vaccine against Aeromonas veronii TH0426 in koi, Cyprinus carpio. Fish and Shellfish Immunology, 81: 176–181, DOI: https://doi.org/10.1016/j.fsi.2018.07.004.
Sun, X., Jin, P., Liu, Q., Wang, Q., Zhang, Y., and Liu, X., 2020. A CpG-riched plasmid as vaccine adjuvant reduce antigen dose of an inactivated Vibrio anguillarum vaccine in turbot (Scophthalmus maximus L.). Fish and Shellfish Immunology, 98: 312–317, DOI: https://doi.org/10.1016/j.fsi.2020.01.031.
Taju, G., Madan, N., Abdul Majeed, S., Kumar, T. R., Thamizhvanan, S., Otta, S. K., et al., 2015. Immune responses of whiteleg shrimp, Litopenaeus vannamei (Boone, 1931), to bacterially expressed dsRNA specific to VP28 gene of white spot syndrome virus. Journal of Fish Diseases, 38(5): 451–465, DOI: https://doi.org/10.1111/jfd.12256.
Tang, L., Kang, H., Duan, K., Guo, M., and Liu, M., 2016. Effects of three types of inactivation agents on the antibody response and immune protection of inactivated IHNV vaccine in rainbow trout. Viral Immunology, 29(7): 430, DOI: https://doi.org/10.1089/vim.2016.0035.
Thompson, F. L., Gevers, D., Thompson, C. C., Dawyndt, P., Naser, S., Hoste, B., et al., 2005. Phylogeny and molecular identification of vibrios on the basis of multilocus sequence analysis. Applied Environmental Microbiology, 71(9): 5107–5115, DOI: https://doi.org/10.1128/AEM.71.9.5107-5115.2005.
Torres, R. J., Cesar, A., Pereira, C., Choueri, R. B., Abessa, D., Nascimento, M. D., et al., 2012. Bioaccumulation of polycyclic aromatic hydrocarbons and mercury in oysters (Crassostrea rhizophorae) from two Brazilian estuarine zones. International Journal of Oceanography, 2012: 1–8, DOI: https://doi.org/10.1155/2012/838320.
Wang, C., Zhang, X. H., Jia, A., Chen, J., and Austin, B., 2008. Identification of immunity-related genes from kidney and spleen of turbot, Psetta maxima (L.), by suppression subtractive hybridization following challenge with Vibrio harveyi. Journal of Fish Diseases, 31(7): 505–514, DOI: https://doi.org/10.1111/j.1365-2761.2008.00914.x.
Wang, P., Lu, Y. Q., Wen, Y., Yu, D. Y., Ge, L., Dong, W. R., et al., 2013. IL-16 induces intestinal inflammation via PepT1 upregulation in a pufferfish model: New insights into the molecular mechanism of inflammatory bowel disease. Journal of Immunology, 191(3): 1413–1427, DOI: https://doi.org/10.4049/jimmunol.1202598.
Xu, W. T., Jiao, C. L., Bao, P. C., Liu, Q., Wang, P. B., Zhang, R. L., et al., 2019. Efficacy of Montanide™ ISA 763 A VG as aquatic adjuvant administrated with an inactivated Vibrio harveyi vaccine in turbot (Scophthalmus maximus L.). Fish and Shellfish Immunology, 84: 56–61, DOI: https://doi.org/10.1016/j.fsi.2018.09.024.
Yan, Y., Chen, Y., Shuang, H., and Ji, C., 2018. Protection against Vibrio alginolyticus in pearl gentian grouper (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatu) immunized with an acfA-deletion live attenuated vaccine. Fish and Shellfish Immunology, 86: 875–881, DOI: https://doi.org/10.1016/j.fsi.2018.12.030.
Yuan, X. Y., Liu, W. B., Liang, C., Sun, C. X., Xue, Y. F., Wan, Z. D., et al., 2017. Effects of partial replacement of fish meal by yeast hydrolysate on complement system and stress resistance in juvenile Jian carp (Cyprinus carpio var. Jian). Fish and Shellfish Immunology, 67: 312–321, DOI: https://doi.org/10.1016/j.fsi.2017.06.028.
Zhang, J., Fu, X., Zhang, Y., Zhu, W., Zhou, Y., Yuan, G., et al., 2019. Chitosan and anisodamine improve the immune efficacy of inactivated infectious spleen and kidney necrosis virus vaccine in Siniperca chuatsi. Fish and Shellfish Immunology, 89: 52–60, DOI: https://doi.org/10.1016/j.fsi.2019.03.040.
Zhang, Z., Yu, Y. X., Wang, Y. G., Liu, X., Wang, L. F., Zhang, H., et al., 2020. Correction to: Complete genome analysis of a virulent Vibrio scophthalmi strain VSc190401 isolated from diseased marine fish half-smooth tongue sole, Cynoglossus semilaevis. BMC Microbiology, 20(1): 375, DOI: https://doi.org/10.1186/s12866-020-02067-0.
Zhou, W., Duan, Z., Yang, B., and Xiao, C., 2017. The effective regulation of pro- and anti-inflammatory cytokines induced by combination of PA-MSHA and BPIFB1 in initiation of innate immune responses. Open Medicine (Wars), 12: 299–307, DOI: https://doi.org/10.1515/med-2017-0044.
Acknowledgements
This work was supported by the Fish Innovation Team of Shandong Agriculture Research System (No. SDAIT-12-06), the Aquatic Animal Immunologic Agents Engineering Research Center of Shandong Province, the Qingdao Agricultural University Doctoral Start-Up Fund (6631122 030), the National Natural Science Foundation of China (No. 32002421), the Advanced Talents Foundation of QAU (No. 6651118016), the Natural Science Foundation of Shandong Province (No. ZR2019BC009), the ‘First-Class Fishery Discipline’ program of Shandong Province, the special top talent plan ‘One Thing One Decision (Yi Shi Yi Yi)’, and the Key Research and Development Program in Shandong Province (No. 2018YFJH0703), Breeding Plan of Shandong Provincial Qingchuang Research Team (2019).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhou, S., Zheng, X., Ding, Y. et al. Immuno-Protective Efficiency of the Bivalent Inactivated Vaccine Against Vibrio scophthalmi and Aeromonas salmonicida Infections in Turbot (Scophthalmus maximus L.). J. Ocean Univ. China 22, 1079–1086 (2023). https://doi.org/10.1007/s11802-023-5455-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11802-023-5455-2