杨惠婕,张玉蕾,刘碧洪,李长玲,黄翔鹄

• 1:广东海洋大学水产学院

摘要(Abstract)：

【目的】研究太平洋亚历山大藻（Alexandrium pacificum）对凡纳滨对虾（Litopenaeus vannamei）的存活率、抗氧化能力以及组织结构的影响。【方法】采用浸浴法，将凡纳滨对虾暴露于太平洋亚历山大藻破碎液中72 h，检测肝胰腺中丙二醛（MDA）、谷胱甘肽（GSH）含量以及超氧化物歧化酶（SOD）、谷胱甘肽—S转移酶（GST）、碱性磷酸酶（AKP）、多酚氧化酶（PPO）等抗氧化酶的活力。对有毒藻浸浴处理和正常海水中对虾的肠道、肝胰腺、鳃、肌肉和神经组织的HE染色石蜡切片进行观察。【结果】太平洋亚历山大藻浸浴处理对凡纳滨对虾的存活率有显著影响（P <0.01）。与对照组相比，太平洋亚历山大藻会诱导对虾肝胰腺的MDA含量增加，导致肝胰腺发生脂质过氧化；同时，诱导GST、GSH、SOD、AKP和PPO的活力增强以清除多余氧自由基，抑制脂质过氧化作用。活性氧（ROS）过多会导致对虾氧化-抗氧化系统的平衡受损，72 h时，实验组对虾肝胰腺的GSH含量显著低于对照组（P <0.01）,SOD、GST以及AKP活力也显著低于对照组（P <0.01）。石蜡切片观察发现，太平洋亚历山大藻引起了对虾肠道、肝胰腺、鳃、肌肉和神经组织细胞肿胀、空泡化等病理变化。【结论】急性暴露于太平洋亚历山大藻会引起对虾重要组织和器官病变，诱导对虾肝胰脏抗氧化酶的改变，这将有助于解释甲壳类动物在太平洋亚历山大藻胁迫下存在的解毒机制。

关键词(KeyWords)： 凡纳滨对虾;太平洋亚历山大藻;抗氧化酶活力;组织病理学

Abstract:

Keywords:

基金项目(Foundation): 广东海洋大学科研启动经费资助项目（060302022102）

作者(Author): 杨惠婕,张玉蕾,刘碧洪,李长玲,黄翔鹄

参考文献(References)：

• [1] UMETSU S, KANDA M, IMAI I, et al. Questiomycins,algicidal compounds produced by the marine bacterium Alteromonas sp. D and their production cue[J]. Molecules(Basel, Switzerland), 2019, 24(24):4522.
• [2] YUSSOF F N, MAAN N, MD REBA M N. LSTM networks to improve the prediction of harmful algal blooms in the west coast of Sabah[J]. International Journal of Environmental Research and Public Health, 2021, 18(14):7650.
• [3] ZHANG Y, YU Z M, SONG X X, et al. Effects of modified clay used for the control of harmful algal blooms on Alexandrium pacificum cysts[J]. Harmful Algae, 2018, 72:36-45.
• [4] SELLNER K G, DOUCETTE G J, KIRKPATRICK G J.Harmful algal blooms:causes, impacts and detection[J].Journal of Industrial Microbiology and Biotechnology,2003, 30(7):383-406.
• [5] INDECK K L, SIMARD P, GOWANS S, et al. A severe red tide(Tampa Bay, 2005)causes an anomalous decrease in biological sound[J]. Royal Society Open Science, 2015, 2(9):150337.
• [6] JOHN U, LITAKER R W, MONTRESOR M, et al. Formal revision of the Alexandrium tamarense species complex(Dinophyceae)taxonomy:the introduction of five species with emphasis on molecular-based(rDNA)classification[J]. Protist, 2014, 165(6):779-804.
• [7] ZOU C, YE R M, ZHENG J W, et al. Molecular phylogeny and PSP toxin profile of the Alexandrium tamarense species complex along the coast of China[J]. Marine Pollution Bulletin, 2014, 89(1/2):209-219.
• [8] FERTOUNA-BELLAKHAL M, DHIB A, FATHALLI A, et al.Alexandrium pacificum Litaker sp. nov(Group IV):resting cyst distribution and toxin profile of vegetative cells in Bizerte Lagoon(Tunisia, Southern Mediterranean Sea)[J]. Harmful Algae, 2015, 48:69-82.
• [9] TAYLOR F R V. Taxonomy of harmful dinoflagellates[J].Manual on harmful marine microalgae, 2003:389-432.
• [10] PéREZ LINARES J, OCHOA J L, GAGO MARTíNEZ A. Retention and tissue damage of PSP and NSP toxins in shrimp:is cultured shrimp a potential vector of toxins to human population?[J]. Toxicon, 2009, 53(2):185-195.
• [11] ROBINEAU B, GAGNéJ A, FORTIER L, et al. Potential impact of a toxic dinoflagellate(Alexandrium excavatum)bloom on survival of fish and crustacean larvae[J]. Marine Biology, 1991, 108(2):293-301.
• [12] PéREZ-LINARES J, OCHOA J L, GAGO-MARTíNEZ A. Effect of PSP toxins in white leg shrimp Litopenaeus vannamei Boone, 1931[J]. Journal of Food Science, 2008,73(4):T69-T73.
• [13] SABATINI S E, BRENA B M, LUQUET C M, et al.Microcystin accumulation and antioxidant responses in the freshwater clam Diplodon chilensis patagonicus upon subchronic exposure to toxic Microcystis aeruginosa[J].Ecotoxicology and Environmental Safety, 2011, 74(5):1188-1194.
• [14] ESTRADA N, DE JESúS ROMERO M, CAMPACóRDOVA A, et al. Effects of the toxic dinoflagellate,Gymnodinium catenatum on hydrolytic and antioxidant enzymes, in tissues of the giant lions-paw scallop Nodipecten subnodosus[J]. Comparative Biochemistry and Physiology Part C:Toxicology&Pharmacology, 2007, 146(4):502-510.
• [15] QIU J B, MA F F, FAN H, et al. Effects of feeding Alexandrium tamarense, a paralytic shellfish toxin producer, on antioxidant enzymes in scallops(Patinopecten yessoensis)and mussels(Mytilus galloprovincialis)[J]. Aquaculture,2013, 396/397/398/399:76-81.
• [16]李磊,刘金殿,黄士林,等.基于综合生物标志物响应评价苯并芘对脊尾白虾(Exopalaemon carinicauda)的毒性效应[J].海洋与湖沼, 2018, 49(2):444-451.
• [17]王芸,李健,李吉涛,等. pH胁迫对中国明对虾抗氧化系统酶活力及基因表达的影响[J].中国水产科学, 2011, 18(3):556-564
• [18]梁忠秀,李健,任海,等.塔玛亚历山大藻对中国明对虾鳃组织的氧化胁迫和对Caspase基因(FcCasp)表达的影响[J].中国水产科学, 2014, 21(1):153-160.
• [19]颜天,谭志军,于仁诚,等.塔玛亚历山大藻对鲈鱼幼鱼毒性效应研究[J].环境科学学报, 2002, 22(6):749-753.
• [20] YAN T, ZHOU M J, FU M, et al. Effects of the dinoflagellate Alexandrium tamarense on early development of the scallop Argopecten irradians concentricus[J]. Aquaculture, 2003,217(1/2/3/4):167-178.
• [21]朱兴彪,郑玉忠,谢丽玲,等.有害赤潮藻种塔玛亚历山大藻的研究进展[J].生物技术进展, 2014, 4(6):421-428
• [22]邓光,李夜光,胡鸿钧,等.温度、光照和pH值对锥状斯氏藻和塔玛亚历山大藻光合作用的影响及光周期对其生长速率和生物量的影响[J].武汉植物学研究, 2004, 22(2):129-134.
• [23]江天久,黄伟建,王朝晖,等.几种环境因子对塔玛亚历山大藻(大鹏湾株)生长及其藻毒力影响[J].应用与环境生物学报, 2000, 6(2):151-154.
• [24] GEHRINGER M M, SHEPHARD E G, DOWNING T G,et al. An investigation into the detoxification of microcystinLR by the glutathione pathway in Balb/c mice[J]. The International Journal of Biochemistry&Cell Biology,2004, 36(5):931-941.
• [25]谭志军,颜天,周名江,等.塔玛亚历山大藻对黑褐新糠虾存活、生长以及种群繁殖的影响[J].生态学报, 2002,22(10):1635-1639
• [26]王雪虹,马嵩.太平洋亚历山大藻对凡纳滨对虾幼体毒性效应研究[J].福建师范大学学报(自然科学版),2007,23(3):58~63.
• [27] SU H M, LIAO I C, CHIANG Y M. Mass mortality of prawn caused by Alexandrium blooming in a culture pond in southern Taiwan[J]. Toxic Phytoplankton Blooms in the sea. Amsterdam:Elsevier Science Publishers B V, 1993.329-333
• [28] S?DERH?LL K, CERENIUS L. Role of the prophenoloxidase-activating system in invertebrate immunity[J].Current Opinion in Immunology, 1998, 10(1):23-28.
• [29] ZHOU J, WANG W N, WANG A L, et al. Glutathione S-transferase in the white shrimp Litopenaeus vannamei:characterization and regulation under pH stress[J].Comparative Biochemistry and Physiology Part C:Toxicology&Pharmacology, 2009, 150(2):224-230.
• [30] VASSEUR P, COSSU-LEGUILLE C. Biomarkers and community indices as complementary tools for environmental safety[J]. Environment International, 2003, 28(8):711-717.
• [31] HALLIWELL B, GUTTERIDGE J M C. Free radicals in biology and medicine[M]. Oxford university press, USA,2015.
• [32] LIVINGSTONE D R. Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms[J]. Marine Pollution Bulletin, 2001, 42(8):656-666.
• [33] STEIN J E, COLLIER T K, REICHERT W L, et al. Bioindicators of contaminant exposure and sublethal effects:studies with benthic fish in puget sound, Washington[J].Environmental Toxicology and Chemistry, 1992, 11(5):701-714.
• [34] PRIETO A I, PICHARDO S, JOSá, et al. Time-dependent oxidative stress responses after acute exposure to toxic cyanobacterial cells containing microcystins in tilapia fish(Oreochromis niloticus)under laboratory conditions[J].Aquatic Toxicology, 2007, 84(3):337-345.
• [35] TRENZADO C, HIDALGO M C, GARCíA-GALLEGO M, et al. Antioxidant enzymes and lipid peroxidation in sturgeon Acipenser naccarii and trout Oncorhynchus mykiss. A comparative study[J]. Aquaculture, 2006, 254(1/2/3/4):758-767.
• [36] SUN Y H, TANG R, LI D P, et al. Acute effects of microcystins on the transcription of antioxidant enzyme genes in crucian carp Carassius auratus[J]. Environmental Toxicology, 2008, 23(2):145-152.
• [37]向华,张亦清,卫文仲,等.人谷胱甘肽硫转移酶A1在乳酸乳球菌中的表达及活力研究[J].微生物学报, 2000, 40(2):132-138.
• [38]彭佑群,王景周,王学峰.托吡酯对大鼠星形细胞内谷胱甘肽硫转移酶及多药耐受基因的影响[J].中国新药与临床杂志, 2004, 23(5):279-282.
• [39] HAYES S C, LUOMA J B, BOND F W, et al. Acceptance and Commitment Therapy:model, processes and outcomes[J]. Behaviour Research and Therapy, 2006, 44(1):1-25.
• [40] GADAGBUI B K M, JAMES M O. Activities of affinityisolated glutathione S-transferase(GST)from channel catfish whole intestine[J]. Aquatic Toxicology, 2000, 49(1/2):27-37.
• [41] COSTA P R, PEREIRA P, GUILHERM E S, et al.Biotransformation modulation and genotoxicity in white seabream upon exposure to paralytic shellfish toxins produced by Gymnodinium catenatum[J]. Aquatic Toxicology,2012, 106/107:42-47.
• [42] GUBBINS M J, EDDY F B, GALLACHER S, et al. Paralytic shellfish poisoning toxins induce xenobiotic metabolising enzymes in Atlantic salmon(Salmo salar)[J]. Marine Environmental Research, 2000, 50(1/2/3/4/5):479-483.
• [43]张喆,李健,陈萍,等.诺氟沙星对中国明对虾鳃和血清ECOD、APND和GST活力的影响[J].中国水产科学,2012, 19(3):514-520.
• [44]李传慧,夏斌,陈碧鹃,等.胜利原油对半滑舌鳎幼鱼肝脏谷胱甘肽过氧化物酶和谷胱甘肽转硫酶活力的影响[J].海洋环境科学, 2011, 30(6):827-830
• [45]李传慧,陈碧鹃,崔毅,等.胜利原油对半滑舌鳎幼鱼肝脏过氧化物酶、溶菌酶和鳃丝Na+-K+-ATPase活力的影响[J].海洋环境科学, 2011, 30(5):681-685
• [46] OIKAWA H, FUJITA T, SATOMI M, et al. Accumulation of paralytic shellfish poisoning toxins in the edible shore crab Telmessus acutidens[J]. Toxicon, 2002, 40(11):1593-1599.
• [47] OIKAWA H, SATOMI M, WATABE S, et al. Accumulation and depuration rates of paralytic shellfish poisoning toxins in the shore crab Telmessus acutidens by feeding toxic mussels under laboratory controlled conditions[J]. Toxicon,2005, 45(2):163-169.
• [48]江天久,徐轶肖.华贵栉孔扇贝→中国龙虾的麻痹性贝类毒素传递与代谢研究[J].海洋学报, 2006, 28(6):169-175.
• [49]梁忠秀,李健,谭志军,等.塔玛亚历山大藻对中国明对虾肝胰腺及鳃SOD、GST和MDA的影响[J].水产学报,2013, 37(8)1192-1197
• [50] MAZORRA M T, RUBIO J A, BLASCO J. Acid and alkaline phosphatase activities in the clam Scrobicularia plana:kinetic characteristics and effects of heavy metals[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology, 2002, 131(2):241-249.
• [51]代涛,尹志峰,王良友.还原型谷胱甘肽临床应用研究进展[J].承德医学院学报, 2014, 31(5):432-435.
• [52] COSTAP R, PEREIRA P, GUILHERME S, et al.Hydroxybenzoate paralytic shellfish toxins induce transient GST activity depletion and chromosomal damage in white seabream(Diplodus sargus)[J]. Marine Environmental Research, 2012, 79:63-69.
• [53] LIKHITWITAYAWUID K. Stilbenes with tyrosinase inhibitory activity[J]. Current Science, 2008:44-52.
• [54] YIN L Y, HUANG J Q, HUANG W M, et al. Responses of antioxidant system in Arabidopsis thaliana suspension cells to the toxicity of microcystin-RR[J]. Toxicon, 2005,46(8):859-864.
• [55] CASTONGUAY M, LEVASSEUR M, BEAULIEU J L,et al. Accumulation of PSP toxins in Atlantic mackerel:seasonal and ontogenetic variations[J]. Journal of Fish Biology, 1997, 50(6):1203-1213.
• [56] COSTA P R, LAGE S, BARATA M, et al. Uptake, transformation, and elimination kinetics of paralytic shellfish toxins in white seabream(Diplodus sargus)[J]. Marine Biology, 2011, 158(12):2805-2811.
• [57]梁忠秀,李健,李吉涛,等.塔玛亚历山大藻毒素及其对中国对虾的急性毒性[J].中国水产科学, 2014, 21(6):1260-1267.
• [58] EMURA A, MATSUYAMA Y, ODA T. Evidence for the production of a novel proteinaceous hemolytic exotoxin by dinoflagellate Alexandrium taylori[J]. Harmful Algae,2004, 3(1):29-37.