Evaluation of the Oral Immune Effects of Pseudomonas plecoglossicida Immune Stimulating Complexes on Large Yellow Croaker (Larimichthys crocea)
XU Bin-Fu1, CHEN Xiu-Xia1, CHI Hong-Shu1, LI Su-Yi1, ZHENG Zai-Yu1, XU Meng-Ting1, JIANG Qiu-Huan1, ZHENG Wei-Qiang2, LIN Neng-Feng1,*
1 Fujian Provincial Aquatic Disease Prevention and Control Engineering Technology Research Center, Institute of Biotechnology, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China; 2 State Key Laboratory of Mariculture Breeding/Ningde FuFa Fisheries Company Limited, Ningde 352103, China
Abstract:Pseudomonas plecoglossicida is the main pathogen which causes the visceral white spot disease in large yellow croaker (Larimichthys crocea), leading to serious economic loss. Previous studies have shown that immune stimulating complexes (ISCOMs) have good oral immune effects. To evaluate the oral immune effects of P. plecoglossicida immune stimulating complexes (Pp-ISCOMs) on L. crocea, Pp-ISCOMs vaccine was prepared, which presented a cage-like nanoparticle structure. At 21 d post oral vaccination, the expression of immune-related genes in the head-kidney of immunized L. crocea were detected, and the immune serum titer was measured using agglutination tests. The immune protective effects of Pp-ISCOMs were then determined by challenge assay with P. plecoglossicida. The results showed that the expression of immune or anti-inflammatory factors such as myeloid differentiation factor 88 (MyD88)(P<0.05), interleukin 2 (IL-2)(P<0.001) in the head kidney of Pp-ISCOMs-immunized L. crocea were significantly higher than those of the inactived-P. plecoglossicida-immunized group (Pp) or the blank control group (CK); and the agglutination titer of large yellow croaker serum (21 d post immunization) was 22. After a P. plecoglossicida attack with a dose of 5×102 CFU/g by intraperitoneal injection, the relative percent survival (RPS) of L. crocea vaccinated by Pp-ISCOMs and Pp were estimated to be 73.3% and 40% respectively. The results indicate that the Pp-ISCOMs could improve the resistance of L. crocea to P. plecoglossicida, and suggested a promising prospect for application. This study provides technical support for the prevention and control by immune to visceral white spot disease in L. crocea.
[1] 陈秀霞, 黄殿盛, 柯翎, 等. 2013. BSA-ISCOMs对大黄鱼免疫机能及抗病力的影响[J]. 福建农业学报, 28(3): 201-205. (Chen X X, Huang D S, Ke L, et al.2013. Effects of BSA-ISCOMs on immunity response and disease resistance of Pseudosciaena crocea[J]. Fujian Journal of Agricultural Science, 28(3): 201-205.) [2] 陈秀霞, 池洪树, 许斌福, 等. 2022. 三七总皂苷对大黄鱼抗变形假单胞菌感染能力的影响[J]. 大连海洋大学学报, 38(3): 380-387. (Chen X X, Chi H S, Xu B F, et al.2022. Effects of panax notoginseng saponins on the resistance of large yellow croaker Larimichthys crocea against Pseudomonas plecoglossicida[J]. Journal of Dalian Ocean University, 38(3): 380-387.) [3] 董传甫, 林天龙, 龚晖, 等. 2005. 嗜水气单胞菌主要外膜蛋白对欧洲鳗鲡的免疫保护试验[J]. 水生生物学报, 29(3): 285-290. (Dong C F, Lin T L, Gong H, et al.2005. Major outer membrane proteins (MOMP) of Aeromonas hydrophila induced protective immunity to European eel(Anguillia anguillia)[J]. Acta Hydrobiologica Sinica, 29(3): 285-290.) [4] 胡娇, 张飞, 徐晓津, 等. 2014. 大黄鱼(Pseudosciaena crocea)内脏白点病病原分离鉴定与致病性研究[J]. 海洋与湖沼, 45(2): 409-417. (Hu J, Zhang F, Xu X J, et al.2014. Isolation, identification and virulence of the pathogen of White-Spots disease in internal organs of Pseudosciaena crocea[J]. Oceanologia et Limnologia Sinica, 45(2): 409-417.) [5] 霍建强, 赵玲敏, 覃映雪, 等. 2017. 变形假单胞菌灭活疫苗研究[J]. 集美大学学报 (自然科学版), 22(4): 12-19. (Huo J Q, Zhao L M, Qin Y X, et al.2017. Studies on inactivate vaccine of Pseudomonas plecoglossicida[J]. Journal of Jimei University (Natural Science), 22(4): 12-19.) [6] 李娜, 周伟芳, 刘慧敏, 等. 2013. 疫苗佐剂的研究现状和发展趋势[J]. 中国预防兽医学报, 35(1): 81-86. (Li N, Zhou W F, Liu H M, et al.2013. Research status and development trends of vaccine adjuvants[J]. Chinese Journal of Preventive Veterinary Medicine, 35(1): 81-86.) [7] 梁箫, 刘钰珠, 陈珂,等. 2019. 厚壳贻贝MyD88-4基因的生物学特性及其对沙氏弧菌的免疫应答[J]. 水产学报, 43(11): 2347-2358. (Liang X, Liu Y Z, Chen K, et al.2019. Identification of MyD88-4 in Mytilus coruscus and expression changes in response to Vibrio chagasii challenge[J]. Journal of Fisheries of China, 43(11): 2347-2358.) [8] 刘洁, 高风英, 卢迈新, 等. 2020. 尼罗罗非鱼TIRAP基因克隆、组织表达及其在无乳链球菌、脂多糖和聚肌胞苷酸刺激下的免疫应答[J]. 大连海洋大学学报, 35(6): 857-865. (Liu J, Gao F Y, Lu M X, et al.2020. Cloning and expression of TIRAP gene and immune response in Nile tilapia (Oreochromis niloticus) to Streptococcus agalactiae, lipopolysaccharides and polyinosinic polycytidylic acid stress[J]. Journal of Dalian Ocean University, 35(6): 857-865.) [9] 刘年锋, 郭睿, 江小斌, 等. 2016. 福州地区大黄鱼变形假单胞菌的分离与鉴定[J]. 渔业研究, 38(1): 14-20. (Liu N F, Guo R, Jiang X B, et al.2016. Isolation and identification of Pseudomonas plecoglossicida in large yellow croaker (Pseudosciaena crocea) of Fuzhou area[J]. Journal of Fisheries Research, 38(1): 14-20.) [10] 王志平, 韩彦军, 李亚琳, 等. 2014. 斑马鱼个体发育过程中的溶菌酶表达模式及其对LPS处理的应答[J]. 甘肃农业大学学报, 49(6): 14-18. (Wang Z P, Han Y J, Li Y L, et al.2014. Lysozyme expression pattern and its response to LPS treatment during zebratfish developm ent process[J]. Journal of Gansu Agricultural University, 49(6): 14-18.) [11] 许斌福, 陈秀锦, 俞伏松, 等. 2007. 创伤弧菌ISCOM疫苗的免疫学效应研究[J]. 中国人兽共患病学报, 23(11): 1075-1078. (Xu B F, Chen X J, Yu F S, et al.2007. Immunological effect of the ISCOM vaccine against Vibrio vulnificus[J]. Chinese Journal of Zoonoses, 23(11): 1075-1078.) [12] 叶立林, 支海兵. 2001. 免疫刺激复合物(ISCOMs)研究进展[J]. 中国兽药杂志, 35(2): 47-51. (Ye L L, Zhi H B.2001. Research progress in immune stimulation complexes (ISCOMs)[J]. Chinese Journal of Veterinary Medicine, 35(2): 47-51.) [13] 张杰, 郭海杰, 高丽婷, 等. 2017. 杀香鱼假单胞菌Ⅲ型分泌系统转录调控蛋白ExsA突变株的构建及其对大黄鱼的毒力特征分析[J]. 水产学报, 41(4): 613-621. (Zhang J, Guo H J, Gao L T, et al.2017. Construction of ExsA,the transcription regulator of TypeⅢ secretion system mutated Pseudomonas plecoglossicida NB2011, and characterization of the virulence against large yellow croaker (Larimichthys crocea)[J]. Journal of Fisheries of China, 41(4): 613-621.) [14] 张杰, 毛芝娟. 2015. 大黄鱼内脏白点病病原杀香鱼假单胞菌及其毒力因子研究进展[J]. 浙江万里学院学报, 28(6): 69-76, 81. (Zhang J, Mao Z J.2015. Research progress on Pseudomonas plecoglossicida and its virulence factors of large yellow croaker (Pseudosciaena crocea)[J]. Journal of Zhejiang Wanli University, 28(6): 69-76,81.) [15] Chen X X, Chi H S, Xu B F, et al.2019. Oral bovine serum albumin immune-stimulating complexes improve the immune responses and resistance of large yellow croaker (Pseudosciaena crocea, Richardson, 1846) against Vibrio alginolyticus[J]. Aquaculture Research, 50: 2802-2809. [16] Companjen A R, Florack D E A, Bastiaans J H M W, et a1.2005. Development of a cost-effective oral vaccination method against viral disease in fish[J]. Developments in Biologicals, 121: 143-150. [17] Grinde B, Lie Q, Poppe T, et a1.1988. Species and individual variation in lysozyme activity in fish of interest in aquaculture[J]. Aquaculture, 68(4): 299-304. [18] Guo S L, Wang Y, Guan R Z, et a1.2013. Immune effects of a bivalent expressed outer membrane protein to American eels (Anguilla rostrota)[J]. Fish&Shellfish Immunology, 35(2): 213-220. [19] Kawai T, Akira S.2010. The role of pattern-recognition receptors in innate immunity: Update on Toll-like receptors[J]. Nature Immunology, 11: 373-384. [20] Livak K J, Schmittgen T D.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 25(4): 402-408. [21] Morelli A B, Becher D, Koernig S, et al.2012. ISCOMATRIX: A novel adjuvant for use in prophylactic and therapeutic vaccines against infectious diseases[J]. Journal of Medical Microbiology, 61: 935-943. [22] Mu P F, Huo J Y, Li X F, et a1.2022. IL-2 signaling couples the MAPK and mTORC1 axes to promote T cell proliferation and differentiation in teleosts[J]. The Journal of Immunology, 208(7): 1616-1631. [23] Palm R C, Landolt M L, Busch R A.1998. Route of vaccine administration:effects on the specific humoral response in rainbow trout Oncorhynchus mykiss[J]. Diseases of Aquatic Organisms, 33(3): 157-166. [24] Qi W L, Xu W, Zhao L M.2019. Protection against Pseudomonas plecoglossicida in Epinephelus coioides immunized with a cspA1-knock-down live attenuated vaccine[J]. Fish and Shellfish Immunology, 89: 498-504. [25] Samuel P C, Mariana R P, Gustavo M E, et a1.2018. Quillaja brasiliensis saponin-based nanoparticulate adjuvants are capable of triggering early immune responses[J]. Scientific Reports, 8: 13582. [26] Thanusha R, Tilusha M, Amir D, et al.2023. Evaluating the intestinal immunity of Asian seabass (Lates calcarifer, Bloch 1790) following field vaccination using a feed-based oral vaccine[J]. Vaccines, 11(3): 602. [27] Yan L, Jin D, Yang S J, et a1.2022. Pathogenicity of fish pathogen Pseudomonas plecoglossicida and preparation of its inactivated vaccine[J]. Microbial Pathogenesis, 166: 105488. [28] Yao C L, Kong P, Wang Z Y, et a1.2009. Molecular cloning and expression of MyD88 in large yellow croaker, Pseudosciaena crocea[J]. Fish&Shellfish Immunology, 26: 249-255. [29] Zhang M J, Chen X Y, Xue M Y, et a1.2023a. Oral vaccination of largemouth bass (Micropterus salmoides) against Largemouth bass ranavirus (LMBV) using yeast surface display technology[J]. Animals, 13(7): 1183. [30] Zhang W B, Zhu C H, Chi H S, et al.2021. Early immune response in large yellow croaker (Larimichthys crocea) after immunization with oral vaccine[J]. Molecular and Cellular Probes, 56: 1-8. [31] Zhang Y B, Huang Y P, Ding H Y, et al.2023b. A σE-mediated temperature gauge orchestrates type VI secretion system, biofilm formation and cell invasion in pathogen Pseudomonas plecoglossicida[J]. Microbiological Research, 266: 127220.