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Histological Assessment and Transcriptome Analysis Provide Insights into the Toxic Effects of Perfluorooctanoic Acid to Juvenile Half Smooth Tongue Sole Cynoglossus semilaevis

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Journal of Ocean University of China 2023年第6期22卷 1635-1648页

作者： ZHAN Min SHI Kunpeng ZHANG Xue FAN Qingxin XU Qian LIU Xinbao LI Zhujun LIU Hongning XIA Yanting SHA Zhenxia Institute of Aquatic Biotechnology Qingdao UniversityQingdao 266071China Laboratory for Marine Fisheries Science and Food Production Processes Laoshan LaboratoryQingdao 266237China

Perfluorooctanoic acid(PFOA)is a widespread synthetic persistent organic pollutant that may enrich along the food chain and affect the growth,development,reproduction,and lipid metabolism of aquatic organisms,particularly the benthic organisms.How-ever,the toxic effects of PFOA on the half-smooth tongue sole Cynoglossus semilaevis,a commercial benthic fish in China,have rarely been reported.Because juvenile fish are sensitive to environmental pollutants,in the present study,histological assessment and tran-scriptome sequencing were performed to determine the short-term impact of PFOA on juvenile half-smooth tongue soles.Histologi-cal analysis showed that PFOA exposure caused hepatocyte rupture,intestinal villi breakage,increased goblet cell count,and brain ab-normal.Transcriptome results showed that some interesting signaling pathways,such as glycolysis/gluconeogenesis,PPAR signaling pathway and GABAergic synapse signaling pathway,were enriched after PFOA exposure.In addition,some metabolic,immune and neural genes were differentially expressed,which including ependymin,hbb1-like and gad 1,and they were up-regulated after 14 days of exposure.Transcriptome results also indicated that half-smooth tongue sole might improve energy metabolism in response to PFOA toxicity after 7 days of exposure.These findings provide a basis for studying the ecological effects of PFOA on marine benthic fishes.

关键词： Cynoglossus semilaevis histological assessment perfluorooctanoic acid transcriptome analysis toxic effect

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Cluster of differentiation antigens:essential roles in the identification of teleost fish T lymphocytes

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marine Life science & Technology 2022年第3期4卷 303-316页

作者： Hong‑fei Tian Jing Xing Xiao‑qian Tang Heng Chi Xiu‑zhen Sheng Wen‑bin Zhan laboratory of Pathology and Immunology of Aquatic Animals KLMMEFisheries CollegeOcean University of ChinaQingdao 266003China Laboratory for Marine Fisheries Science and Food Production Processes Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China

Cluster of differentiation(CD)antigens are cell surface molecules expressed on leukocytes and other cells associated with the immune system.Antibodies that react with CD antigens are known to be one of the most essential tools for identifying leukocyte subpopulations.T lymphocytes,as an important population of leukocytes,play essential roles in the adaptive immune system.Many of the CD antigens expressed on T lymphocytes are used as surface markers for T lymphocyte classification,including CD3,CD4 and CD8 molecules.In this review,we summarize the recent advances in the identification of CD molecules on T lymphocytes in teleosts,with emphasis on the functions of CD markers in the classification of T lymphocyte subsets.We notice that genes encoding CD3,co-receptors CD4 and CD8 have been cloned in several fish species and antibodies have been developed to study protein expression in morphological and functional contexts.T lymphocytes can be divided into CD4^(+)and CD8^(+)cells discriminated by the expression of CD4 and CD8 molecules in teleost,which are functionally similar to mammalian helper T cells(Th)and cytotoxic T cells(Tc),respectively.Further studies are still needed on the particular characteristics of teleost T cell repertoires and adaptive responses,and results will facilitate the health management and development of vaccines for fish.

关键词： T lymphocytes Surface marker Classification Monoclonal antibody Fish

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Two bicistronic DNA vaccines against Vibrio anguillarum and the immune eff ects on flounder Paralichthys olivaceus

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Journal of Oceanology and Limnology 2022年第2期40卷 786-804页

作者： Hanlin LI Jing XING Xiaoqian TANG Xiuzhen SHENG Heng CHI Wenbin ZHAN laboratory of Pathology and Immunology of Aquatic Animals KLMMEOcean University of ChinaQingdao 266003China Laboratory for Marine Fisheries Science and Food Production Processes Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266071China

Chemokines are cytokines that can promote the activation and migration of immune cells,and increase the recognition of antigen by antigen-presenting cells(APC).Previous studies showed that a DNA vaccine can induce humoral and cellular immune responses of flounder after immunization.To explore the improvement of chemokines on the efficiency of OmpK vaccine,two bicistronic DNA candidate vaccines were constructed and the immune responses they induced in the flounder were investigated by reverse transcription polymerase chain reaction(RT-PCR),indirect immunofl uorescent assay(IFA),H&E staining,fl ow cytometry(FCM),and quantifi cational real-time polymerase chain reaction(qRT-PCR).pBudCE4.1 plasmid as an expression vector,bicistronic DNA vaccines encoding OmpK gene and CC-motif ligand 4 gene(p-OmpK-CCL4),or Ompk gene and CC-motif ligand 19 gene(p-OmpK-CCL19)were successfully constructed.The results showed that two bicistronic DNA vaccines expressed Ompk protein of Vibrio anguillarum and CCL4/CCL19 proteins of fl ounder both in vitro and in vivo.After immunization,a large number of leucocytes in muscle were recruited at the injection site in treatment groups.The constructed vaccines induced signifi cant increases in CD4-1^(+) and CD4-2^(+) T lymphocytes,and sIgM^(+) B lymphocytes in peripheral blood,spleen,and head kidney.The percentage of T lymphocytes peaked on the 14^(th) post-vaccination day whereas that of B lymphocytes peaked in the 6^(th) post-vaccination week.Moreover,the expression profi les of 10 immune-related genes increased in muscles around the injection site,spleen,and head kidney.After the challenge,p-OmpK-CCL4 and p-OmpK-CCL19 conferred a relative percentage survival(RPS)of 74.1%and 63.3%,respectively,higher than p-OmpK alone(40.8%).In conclusion,both CCL4 and CCL19 can improve the protection of p-OmpK via evoking local immune response and then humoral and cellular immunity.CCL4 and CCL19 will be potential molecular adjuvants for use in DNA vaccines.

关键词： Vibrio anguillarum outer membrane protein K bicistronic DNA vaccines CC-motif ligand 4 CC-motif ligand 19 immune response

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Productive Traits and Triploid Rate Stability in Triploidy-Induced ‘Haida No. 2’ Strain of the Pacific Oyster, Crassostrea gigas

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Journal of Ocean University of China 2023年第1期22卷 229-234页

作者： LI Yongguo LI Qi LIU Ye XU Chengxun Key laboratory of Mariculture Ministry of EducationOcean University of ChinaQingdao 266003China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266237China

In order to evaluate the effects of triploidy induction on a selected strain‘Haida No.2’of the Pacific oyster Crassostrea gigas,which is characterized with golden shell color and high growth rate,the growth,survival rate and stability of triploid rate were analyzed at different development stages in the present study.Three different conditions inhibiting the release of polar body Ⅱ or polar body Ⅰ were tested:(A)Cytochalasin-B(CB),0.5mg L^(−1) at 10min post-insemination for 15 min;(B)CB,0.5mg L^(−1)at 15 min postinsemination for 20 min;and(C)CB,0.7mg L^(−1),at 15 min post-insemination for 20 min.The triploidy induction treatments significantly reduced the D-larvae and survival rates at the larvae stage but not at the juvenile and adult stages.Triploid rate dramatically decreased at the larval stage and did not significantly change at the juvenile and adult stages.Regarding the stability of the triploid rate,there was a significant difference between the three treatment groups.Larvae from the treatment A and control groups exhibited higher growth rates in shell height than those from the other two treatment groups at day 27.Triploid juveniles and adults from the treatment A group exhibited a higher wet weight than diploids from the control group and triploids from the other treatment groups.Triploidy induction did not affect the shell color of the progeny.The results obtained in the study demonstrate that triploidy induction has the potential to be used to increase the production of C.gigas variety‘Haida No.2’without modifying its golden shell color.

关键词： Crassostrea gigas selective strain triploidy induction survival growth triploid rate stability

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Non-Targeted Metabolomics Reveals the Metabolic Alterations in Response to Artificial Selective Breeding in the Fast-Growing Strains of Pacific Oyster

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Journal of Ocean University of China 2024年第2期23卷 518-528页

作者： HU Boyang TIAN Yuan LIU Shikai LI Qi Key laboratory of Mariculture Ministry of EducationOcean University of ChinaQingdao 266003China Laboratory for Marine Fisheries Science and Food Production Processes Laoshan LaboratoryQingdao 266237China

Pacific oyster(Crassostrea gigas)is one of the most important mollusks cultured all around the world.Selective breeding programs of Pacific oysters in China is initiated since 2006 and developed the genetically improved strain with fast-growing trait.However,little is known about the metabolic signatures of the fast-growing trait.In the present study,the non-targeted metabolomics was performed to analyze the metabolic signatures of adductor muscle tissue in one-year old Pacific oysters from fast-growing strain and the wild population.A total of 7767 and 10174 valid peaks were extracted and quantified in ESI^(+)and ESI^(−)modes,resulting in 399 and 381 annotated metabolites,respectively.PCA and OPLS-DA revealed that considerable separation among samples from fastgrowing strain and wild population,suggesting the differences in metabolic signatures.Meanwhile,81 significantly different metabolites(SDMs)were identified in the comparisons between fast-growing strain and wild population,based on the strict thresholds.It was found that there were highly correlation and conserved coordination among these SDMs.KEGG enrichment analysis indicated that the SDMs were tightly related to pantothenate and CoA biosynthesis,steroid hormone biosynthesis,riboflavin metabolism,and arginine and proline metabolism.Of them,the CoA biosynthesis and metabolism,affected by pantetheine and pantothenic acid,might be important for the growth of Pacific oysters under artificial selective breeding.The study provides the comprehensive views of metabolic signatures in response to artificially selective breeding,and is helpful to better understand the molecular mechanism of fastgrowing traits in Pacific oysters.

关键词： metabolic signature Pacific oyster artificial selection fast-growing trait

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Transcriptome Analysis Reveals New Insights into the Respiration Metabolism Mechanism of Different Feeding Rations of Sea Cucumber(Apostichopus japonicus)

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Journal of Ocean University of China 2023年第6期22卷 1621-1634页

作者： MEI Yaoping HOU Zhishuai GAO Qinfeng DONG Shuanglin LI Xueqi XU Yuling Key laboratory of Mariculture Ministry of EducationOcean University of ChinaQingdao 266003China Function Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266235China

Sea cucumber(Apostichopus japonicus)is an excellent model for investigating effects of bottom-dwellers on carbon mig-ration and transformation.However,the molecular mechanism of respiratory metabolism process variation caused by feeding rations is poorly understood.In this study,treatment groups set as 1%(about 0.63g),3%,and 7%of total body weight(named F1,F3 and F7 groups respectively).The potential molecular mechanisms behind the functions of respiratory tree and body wall were investigated by RNA-Seq.A total of 52411 expressed genes were identified from 89342 expressed transcripts.The results showed 759,254 and 334 genes were up-regulated,and 334,445 and 992 genes were down-regulated in respiratory tree of F1 vs.F3,F1 vs.F7 and F3 vs.F7,respectively.Meanwhile,2070,1601 and 896 genes were up-regulated,and 1303,1337 and 1144 genes were down-regulated in body wall between F1 vs.F3,F1 vs.F7 and F3 vs.F7,respectively.Differentially expressed genes were enriched in salivary secretion and ECM-receptor interaction pathways in respiratory tree,and in various types of N-glycan biosynthesis,ribosome and sphingolipid metabolism pathways in body wall.These results suggested respiratory tree and body wall were involved in activation of respiratory metabolisms in response to different feeding rations.Our research provided valuable knowledge for physiological differences in res-piratory metabolism.

关键词： transcriptomics Apostichopus japonicus feeding ration respiration metabolism

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Gene duplication and functional divergence of new genes contributed to the polar acclimation of Antarctic green algae

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marine Life science & Technology 2023年第4期5卷 511-524页

作者： Xiaowen Zhang Wentao Han Xiao Fan Yitao Wang Dong Xu Ke Sun Wei Wang Yan Zhang Jian Ma Naihao Ye State Key laboratory of Mariculture Biobreeding and Sustainable Goods Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdao 266071China Laboratory for Marine Fisheries Science and Food Production Processes Laoshan LaboratoryQingdao 266200China

Psychrophilic microalgae successfully survive in the extreme and highly variable polar ecosystems,which represent the energy base of most food webs and play a fundamental role in nutrient cycling.The success of microalgae is rooted in their adaptive evolution.Revealing how they have evolved to thrive in extreme polar environments will help us better understand the origin of life in polar ecosystems.We isolated a psychrophilic unicellular green alga,Microglena sp.YARC,from Antarctic sea ice which has a huge genome.Therefore,we predicted that gene replication may play an important role in its polar adaptive evolution.We found that its protein-coding gene number significantly increased and the duplication time was dated between 37 and 48 million years ago,which is consistent with the formation of the circumpolar Southern Ocean.Most duplicated paralogous genes were enriched in pathways related to photosynthesis,DNA repair,and fatty acid metabolism.Moreover,there were a total of 657 Microglena-specific families,including collagen-like proteins.The divergence in the expression patterns of the duplicated and species-specific genes reflects sub-and neo-functionalization during stress acclimation.Overall,key findings from this study provide new information on how gene duplication and their functional novelty contributed to polar algae adaptation to the highly variable polar environmental conditions.

关键词： Psychrophilic microalgae Adaptive evolution Gene replication

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Isolation and Biochemical Characteristics Analyses of Phenoloxidases (POs) in Three Cultured Mollusk Species

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Journal of Ocean University of China 2022年第2期21卷 465-472页

作者： SONG Xiangdi JIANG Jingwei XING Jing ZHAN Wenbin laboratory of Pathology and Immunology of Aquatic Animals KLMMEOcean University of ChinaQingdao 266003China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266235China

Phenoloxidases(POs)are a group of copper proteins including tyrosinase,catecholase and laccase,which play crucial roles in the innate immune response of mollusks.In this research,POs were studied in cultured mollusk species,including scallop Chla-mys farreri,abalone Haliotis discus hannai and clam Scapharca subcrenata.The POs were isolated from hemocytes using linear-gradient native-PAGE combined with catechol staining.The PO activities and their characters were investigated.The molecular mass of PO in C.farreri was 576 kDa,and it was 228 kDa in H.discus hannai.In S.subcrenata,four POs were detected and their mole-cular masses were 391 kDa,206 kDa,174 kDa and<67 kDa,which were named as 391-PO,206-PO,174-PO and s-PO,respectively.Ki-netic analyses indicated that all of the POs,except for 391-PO had higher affinity to L-DOPA and catechol than to hydroquinone and dopamine.However,all of the POs failed to oxidize tyrosine.The effects of divalent metal ions on POs’activities were assayed,in-cluding Fe^(2+),Mg^(2+),Zn^(2+),Mn^(2+),Cu^(2+)and Ca^(2+)from FeCl_(2),MgSO_(4),ZnSO_(4),MnCl_(2),CuSO_(4)and CaCl_(2).The POs were inhibited by Fe^(2+)at all determined concentrations.Additionally,the inhibition assay showed that all of the POs were inhibited by cysteine,ascorbic acid,sodium sulfite,citric acid,ethylenediaminetetraacetic acid disodium(EDTA)and sodium diethyldithiocarbamate(DETC).The inhibition effects of critric acid and EDTA are dose-dependent.H.discus hannai PO and 391-PO were slightly inhibited by sodium azide,and H.discus hannai PO,391-PO and 174-PO were slightly inhibited by thiourea.In conclusion,the POs in the three cultured mollusks are copper-containing laccase-type phenoloxidases with similar biochemical characteristics even though their molecular masses are different.

关键词： Chlamys farreri Haliotis discus hannai Scapharca subcrenata phenoloxidase divalent metal ions inhibitors

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The impact of natural mortality variations on the performance of management procedures for Spanish mackerel(Scomberomorus niphonius) in the Yellow Sea, China

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Acta Oceanologica Sinica 2018年第8期37卷 21-30页

作者： CHEN Ning ZHANG Chongliang SUN Ming XU Binduo XUE Ying REN Yiping CHEN Yong College of fisheries Ocean University of China Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and Technology School of marine sciences University of Maine

Natural mortality rate（M） is one of the essential parameters in fishery stock assessment, however, the estimation of M is commonly rough and the changes of M due to natural and anthropogenic impacts have long been ignored.The simplification of M estimation and the influence of M variations on the assessment and management of fisheries stocks have been less well understood. This study evaluated the impacts of the changes in natural mortality of Spanish mackerel（Scomberomorus niphonius） on their management strategies with data-limited methods. We tested the performances of a variety of management procedures（MPs） with the variations of M in mackerel stock using diverse estimation methods. The results of management strategies evaluation showed that four management procedures DCAC, SPMSY, cur E75 and minlen Lopt1 were more robust to the changes of M than others; however, their performance were substantially influenced by the significant decrease of M from the 1970s to 2017. Relative population biomass（measure as the probability of B〉0.5 BMSY） increased significantly with the decrease of M, whereas the possibility of overfishing showed remarkable variations across MPs. The decrease of M had minor effects on the long-term yield of cur E75 and minlen Lopt1, and reduced the fluctuation of yield（measure as the probability of AAVY〈15%） for DCAC, SPMSY. In general, the different methods for M estimation showed minor effects on the performance of MPs, whereas the temporal changes of M showed substantial influences. Considering the fishery status of Spanish mackerel in China, we recommended that cur E75 has the best trade-off between fishery resources exploitation and conservation, and we also proposed the potentials and issues in their implementations.

关键词： fishery management uncertainty management strategy evaluation（MSE） data limited method DLMtool

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Response of Yesso scallop Patinopecten yessoensis to acute temperature challenge: physiological and biochemical parameters

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Journal of Oceanology and Limnology 2019年第1期37卷 321-329页

作者： JIANG Weiwei DU Meirong FANG Jianguang GAO Yaping MAO Yuze CHEN Qionglin LIN Fan JIANG Zengjie Key laboratory of Sustainable Development of marine fisheries Ministry of Agriculture Yellow Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Laboratory for Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and Technology

Water temperature is generally considered to be a major factor af fecting the physiological and biochemical activities of marine bivalves. Here, the physiological and biochemical responses of Yesso scallop, Patinopecten yessoensis, to acute water temperature changes in summer were studied. Scallops were transferred directly to a lower temperature( T dec treatment)(from 23°C to 15°C) or to a higher temperature( Ti nc treatment)(from 15°C to 23°C) for 72 h, respectively. Results showed that the oxygen consumption and ammonia-N excretion rates of P. yessoensis decreased signi?cantly in the T dec treatment but increased dramatically at 6 h in the Ti nc treatment( P <0.05). In the T dec treatment, hepatopancreas antioxidant enzyme activities, superoxide dismutase(SOD) and catalase(CAT) activities, increased substantially within 72 h( P <0.05). However, a signi?cant decrease in CAT activity was found at 12 h in the Ti nc treatment( P <0.01).A signi?cant enhancement of acid phosphatase(ACP) activity and malondialdehyde(MDA) content was detected when scallops were acutely exposed to a temperature of 15°C. The levels of Cu/Zn-SOD gene expression in their gills up-regulated signi?cantly in response to acute temperature changes( P <0.01). These data suggest that acute temperature change af fects physiological and biochemical functions, and improve our knowledge of P. yessoensis under conditions of thermal stress.

关键词： Patinopecten yessoensis acute water temperature changes physiology biochemical activities

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