Determination of ED<sub>50</sub> of transgenic cyanobacteria oral vaccine and safety evaluation on <i>Danio rerio</i>

XU Yang; XIE Jingkun; LI Yunhui; SU Yongzheng; HE Peimin; JIA Rui

doi:10.11964/jfc.20200212149

Volume 45 Issue 2

Feb. 2021

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Journal of fisheries of china > 2021 > 45(2): 255-264. > DOI: 10.11964/jfc.20200212149

XU Yang, XIE Jingkun, LI Yunhui, SU Yongzheng, HE Peimin, JIA Rui. Determination of ED₅₀ of transgenic cyanobacteria oral vaccine and safety evaluation on Danio rerio[J]. Journal of fisheries of china, 2021, 45(2): 255-264. DOI: 10.11964/jfc.20200212149

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Determination of ED₅₀ of transgenic cyanobacteria oral vaccine and safety evaluation on Danio rerio

1.
College of Marine Ecology and Environment, Shanghai Ocean University, Shanghai 201306, China
2.
Water Environment & Ecology Engineering Research Center of Shanghai Institute of Higher Education, Shanghai Ocean University, Shanghai 201306, China

Funds: National Key Research and Development Project (2019YFC0312604); Shanghai Agriculture Science and Technology Innovation Project (2017, 1-13); National High Technology Research and Development Program of China (2014AA093506); Student Innovation Training Project (S201910264057)

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Corresponding author:
JIA Rui. E-mail: rjia@shou.edu.cn
Received Date: February 03, 2020
Revised Date: June 18, 2020
Available Online: August 12, 2020
Published Date: January 31, 2021

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Abstract

Abstract

Nowadays, in aquaculture, the popularity of antibiotics and the increase in the use of other drugs have made it easier for residual antibiotics and drugs to invade natural water bodies. The environmental effects of antibiotics in the ecosystem have caused more and more pollution to the water environment and threatened the survival of aquatic organisms. After strict regulation and control of the use of antibiotics, the use of genetically modified vaccines that can replace antibiotics has become more and more common. Cyanobacteria is a natural bait for shrimp seedlings. Directly feeding Litopenaeus vannamei with Synechococcus-expressedvp28 to control WSSV can play a role of “medicinal and food homology”. However, the problem facing the current application of transgenic cyanobacteria as a subunit vaccine is the safety of its environmental release. Danio rerio is one of the most commonly used model creatures. In the previous Synechococcus-expressedvp28 oral vaccine efficacy experiment, the feasibility of the oral vaccine was verified, showing that its efficacy and safety can be trusted. This environmental release experiment was built on this basis. In this study, Synechococcus-expressedvp28 were administered to L. vannamei to verify the effect of its dose gradient against WSSV and to determine its half effective dose (ED₅₀). After feeding D. rerio on the basis of this ED₅₀, the effects of the Synechococcus-expressed vp28 on aquatic organisms and water environment were explored by measuring the enzymatic indicators in D. rerio, observing the tissue sections, and the changes in nitrogen and phosphorus in the aquaculture water. The results of the study showed that with the increase of the dose of Synechococcus-expressedvp28 oral vaccine, the anti-WSSV efficacy of L. vannamei also increased and the measured ED₅₀ was 0.027 g. Feeding D. rerio by freeze-dried wild-type and transgenic Synechococcus with the shelled-out Artemia for 15 days, the body length and color of D. rerio were not significantly different. The CAT in the transgenic group was significantly lower than that blank and wild-type groups. There was no significant difference in SOD and POD between wild-type and transgenic groups. There was no significant difference in the liver and heart of D. rerio through the tissue section. The ammonia nitrogen in wild-type and transgenic groups was lower than that in the blank group, and there was no significant difference between total nitrogen and total phosphorus. The results of this study further showed that the Synechococcus-expressed vp28 can effectively enhance the anti-WSSV ability of L.vannamei, and as its dose increases, the disease resistance has a more significant expression. In the current experimental results, there is no obvious toxicological effect on D. rerio, and after feeding with Synechococcus-expressed vp28, it does not show a significant effect on the quality of aquaculture water. This shows that this subunit vaccine has less impact on aquatic organisms, and can provide a basis and more possibilities for subsequent industrial-scale applications and development.
- transgenic cyanobacteria,
- Danio rerio,
- vp28,
- median effective dose (ED₅₀),
- safety

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References

[1]	Flegel T W. Major viral diseases of the black tiger prawn (Penaeus monodon) in Thailand[J]. World Journal of Microbiology and Biotechnology, 1997, 13(4): 433-442. doi: 10.1023/A:1018580301578
[2]	殷嵘, 郭媛媛, 韦章良, 等. 不同途径感染白斑综合征病毒(WSSV)对日本沼虾的致病性[J]. 生物工程学报, 2017, 33(6): 946-956. Yin R, Guo Y Y, Wei Z L, et al. Pathogenicity of white-spot syndrome virus in Macrobrachium nipponensis via different infection routes[J]. Chinese Journal of Biotechnology, 2017, 33(6): 946-956(in Chinese).
[3]	傅玲琳, 帅江冰, 许梓荣. 对虾白斑综合征病毒的分子生物学研究进展[J]. 中国兽医杂志, 2005, 41(4): 42-44. doi: 10.3969/j.issn.0529-6005.2005.04.016 Fu L L, Shuai J B, Xu Z R. Recent advances in the molecular biology of white spot syndrome virus of prawns[J]. Chinese Journal of Veterinary Medicine, 2005, 41(4): 42-44(in Chinese). doi: 10.3969/j.issn.0529-6005.2005.04.016
[4]	衣启麟, 王玲玲, 宋林生. 对虾白斑综合征及其免疫防控[J]. 生命科学, 2014, 26(9): 903-911. Yi Q L, W L L, Song L S. White spot syndrome in shrimp and its immunological prevention and control[J]. Chinese Bulletin of Life Sciences, 2014, 26(9): 903-911(in Chinese).
[5]	Li Z J, Lin Q S, Chen J, et al. Shotgun identification of the structural proteome of shrimp white spot syndrome virus and iTRAQ differentiation of envelope and nucleocapsid subproteomes[J]. Molecular & Cellular Proteomics, 2007, 6(9): 1609-1620.
[6]	Sánchez-Paz A. White spot syndrome virus: an overview on an emergent concern[J]. Veterinary Research, 2010, 41(6): 43. doi: 10.1051/vetres/2010015
[7]	Xie X X, Xu L M, Yang F. Proteomic analysis of the major envelope and nucleocapsid proteins of white spot syndrome virus[J]. Journal of Virology, 2006, 80(21): 10615-10623. doi: 10.1128/JVI.01452-06
[8]	Zhang H D, Kolb F A, Brondani V, et al. Human Dicer preferentially cleaves dsRNAs at their termini without a requirement for ATP[J]. The EMBO Journal, 2002, 21(21): 5875-5885. doi: 10.1093/emboj/cdf582
[9]	王晓雯. 凡纳滨对虾与WSSV侵染相关的细胞与基因功能分析[D]. 青岛: 中国海洋大学, 2014. Wang X W. Functional analysis of the cells and the gene related with WSSV infection in Litopenaeus vannamei[D]. Qingdao: Ocean University of China, 2014 (in Chinese).
[10]	Li Z C, Xu L M, Li F, et al. Analysis of white spot syndrome virus envelope protein complexome by two-dimensional blue native/SDS PAGE combined with mass spectrometry[J]. Archives of Virology, 2011, 156(7): 1125-1135. doi: 10.1007/s00705-011-0954-7
[11]	席超, 王春梅, 施定基. 蓝藻基因工程应用研究进展[J]. 中国生物工程杂志, 2010, 30(3): 105-111. Xi C, Wang C M, Shi D J. Advances on Cyanobacteria genetic engineering applications[J].China Biotechnology, 2010, 30(3): 105-111 (in Chinese).
[12]	贾睿, 朱婵, 庄旻敏, 等. 白斑综合征病毒囊膜蛋白VP19及VP28的研究进展[J]. 水产学报, 2020, 44(5): 870-880. Jia R, Zhu C, Zhuang M M, et al. Review of research progress on white spot syndrome virus (WSSV) envelope protein VP19 and VP28[J]. Journal of Fisheries of China, 2020, 44(5): 870-880(in Chinese).
[13]	郭媛媛, 殷嵘, 施定基, 等. 转vp28蓝藻口服剂对凡纳滨对虾抗白斑综合征病毒能力及免疫反应的影响[J]. 水产学报, 2017, 41(9): 1473-1485. Guo Y Y, Yin R, Shi D J, et al. Effect of Anabaena-expressed vp28 against white spot syndrome virus and related immune response in Litopenaeus vannamei[J]. Journal of Fisheries of China, 2017, 41(9): 1473-1485(in Chinese).
[14]	谢菲, 陈晗, 李建明, 等. 斑马鱼在肿瘤研究中的应用[J]. 中国热带医学, 2009, 9(6): 1161-1164. Xie F, Chen H, Li J M, et al. Application of zebrafish in cancer research[J]. China Tropical Medicine, 2009, 9(6): 1161-1164(in Chinese).
[15]	Gao Y J, Zhu H J, Chen Y, et al. Safety assessment of Bacillus thuringiensis insecticidal proteins Cry1C and Cry2A with a zebrafish embryotoxicity testth a zebrafish embryotoxicity test[J]. Journal of Agricultural and Food Chemistry, 2018, 66(17): 4336-4344. doi: 10.1021/acs.jafc.8b01070
[16]	董姗姗, 章嫡妮, 张振华, 等. 转mCry1Ac基因玉米BT799对斑马鱼的生态毒理学效应[J]. 应用生态学报, 2019, 30(8): 2845-2853. Dong S S, Zhang D N, Zhang Z H, et al. Ecotoxicological effects of transgenic mCry1Ac maize (BT799) on zebrafish[J]. Chinese Journal of Applied Ecology, 2019, 30(8): 2845-2853(in Chinese).
[17]	施定基, 王永旭, 方昭希. 固定化对多变鱼腥藻突变种及其野生型的光合、生长、固氮和泌氨的影响[J]. 植物学报, 1991, 33(5): 335-342. Shi D J, Wang Y X, Fang Z X. The effects of immobilization on photosynthesis, growth, nitrogen fixation and ammonium ion excretion of mutant and wild type cells of Anabaena variabilis[J]. Acta Botanica Sinica, 1991, 33(5): 335-342(in Chinese).
[18]	GB 11893-89, 水质—总磷的测定——钼酸铵分光光度法[S]. 北京: 中国环境科学出版社, 1989. GB 11893-89, Water quality - determination of total phosphorus - ammonium molybdate spectrophotometric method[S]. Beijing: China Environmental Science Press, 1989.
[19]	HJ 636-2012, 水质−总氮的测定——碱性过硫酸钾消解紫外分光光度法[S]. 北京: 中国环境科学出版社, 2012. HJ 636-2012, Water quality-determination of total nitrogen - alkaline potassium persulfate digestion UV spectrophotometric method[S]. Beijing: China Environmental Science Press, 2012.
[20]	HJ536-2009, 水质−氨氮的测定——水杨酸分光光度法[S]. 北京: 中国环境科学出版社, 2010. HJ536-2009, Water quality-determination of ammonia nitrogen - salicylic acid spectrophotometry[S]. Beijing: China Environmental Science Press, 2010.
[21]	封琦, 朱光来, 王建国, 等. 氨氮对中华鳑鲏的急性毒性及2种代谢酶活性的影响[J]. 淡水渔业, 2018, 48(1): 91-96. doi: 10.3969/j.issn.1000-6907.2018.01.015 Feng Q, Zhu G L, Wang J G, et al. Acutetoxicity of ammonia nitrogen to Rhodeus sinensis and its effects on two metabolism enzymes[J]. Freshwater Fisheries, 2018, 48(1): 91-96(in Chinese). doi: 10.3969/j.issn.1000-6907.2018.01.015
[22]	Subhadra B, Hurwitz I, Fieck A, et al. Development of paratransgenic Artemia as a platform for control of infectious diseases in shrimp mariculture[J]. Journal of Applied Microbiology, 2010, 108(3): 831-840. doi: 10.1111/j.1365-2672.2009.04479.x
[23]	陈昌生, 王淑红, 纪德华, 等. 氨氮对九孔鲍过氧化氢酶和超氧化物歧化酶活力的影响[J]. 上海海洋大学学报, 2001, 10(3): 218-222. doi: 10.3969/j.issn.1004-7271.2001.03.005 Chen C S, Wang S H, Ji D H, et al. Effects of ammonia-N on activities of CAT and SOD in Haliotis diversicolor supertexta[J]. Journal of Shanghai Ocean University, 2001, 10(3): 218-222(in Chinese). doi: 10.3969/j.issn.1004-7271.2001.03.005
[24]	韩力强, 康现江, 李双石, 等. 氨氮对斑马鱼2种代谢酶类的影响[J]. 河北大学学报(自然科学版), 2005, 25(2): 179-184. Han L Q, Kang X J, Li S S, et al. Effects of ammonia nitrogen on metabolic enzymes in Brachyclanio rerio[J]. Journal of Hebei University (Natural Science Edition), 2005, 25(2): 179-184(in Chinese).
[25]	许星鸿, 张雁秋, 阎斌伦, 等. 氨氮胁迫对日本蟳免疫生理指标及器官结构的影响[J]. 生态学报, 2014, 34(14): 3885-3894. Xu X H, Zhang Y Q, Yan B L, et al. Effects of ammonia-N stress on immunity-related indicators and histological structure of some organs of the marine crab, Charybdis japonica (A. Milne-Edwards)[J]. Acta Ecologica Sinica, 2014, 34(14): 3885-3894(in Chinese).
[26]	邱德全, 杨士平, 邱明生. 氨氮促使携带白斑综合征病毒凡纳滨对虾发病及其血细胞、酚氧化酶和过氧化氢酶变化[J]. 渔业现代化, 2007, 34(1): 36-39. doi: 10.3969/j.issn.1007-9580.2007.01.013 Qiu D Q, Yang S P, Qiu M S. Effect of ammonia on disease of white shrimp, Litopenaeus vannamei with WSSV[J]. Fishery Modernization, 2007, 34(1): 36-39(in Chinese). doi: 10.3969/j.issn.1007-9580.2007.01.013
[27]	姜玮. 基于模式生物斑马鱼的“藻戟遂芫俱战草”急性毒性相互作用研究[D]. 南京: 南京中医药大学, 2012. Jiang W. Study on acute toxicity and interaction of the “Haizao, Daji, Gansui and Yuanhua Reverse Gancao” based on zebrafish[D]. Nanjing: Nanjing University of Chinese Medicine, 2012 (in Chinese).
[28]	殷嵘, 顾娟, 施定基, 等. pH法防止转基因蓝藻向环境释放[J]. 生物技术, 2017, 27(6): 592-600. Yin R, Gu J, Shi D J, et al. pH method to prevent the release of transgenic Anabaena sp. into environment[J]. Biotechnology, 2017, 27(6): 592-600(in Chinese).
[29]	吴倩萍, 刘海萍, 顾娟, 等. 冻融法控制制药用转基因蓝藻在环境中释放[J]. 中国海洋药物, 2018, 37(2): 25-31. Wu Q P, Liu H P, Gu J, et al. Controlling environmental release of transgenic cyanobacteria used to prepare recombinant drug by freezing-thawing way inhibitor metyrapone[J]. Chinese Journal of Marine Drugs, 2018, 37(2): 25-31(in Chinese).

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Determination of ED₅₀ of transgenic cyanobacteria oral vaccine and safety evaluation on Danio rerio

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Determination of ED₅₀ of transgenic cyanobacteria oral vaccine and safety evaluation on Danio rerio