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Optimization of Protectant, Salinity and Freezing Condition for Freeze-Drying Preservation of Edwardsiella tarda

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Journal of Ocean University of China 2017年第5期16卷 831-839页

作者： YU Yongxiang ZHANG Zheng WANG Yingeng LIAO Meijie LI Bin XUE Liangyi College of marine Sciences Ningbo University Laboratory for marine fisheries Science and Food Production Processes Yellow Sea Fisheries Research InstituteChinese Academy of Fishery Sciences

Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose(8.0%), skim milk(12.0%), sodium citrate(2.0%), serum(12.0%) and PVP(2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of Na Cl was 10.0, 30.0 and between 5.0 and 10.0 g L^(-1) for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at-80℃ or-40℃ for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate(79.63%–82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy(SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda.

关键词： tarda pathogen citrate preservation preserving contained preparing thick additive suspended

Influence of White Spot Syndrome Virus Infection on Hepatopancreas Gene Expression of ‘Huanghai No.2' Shrimp(Fenneropenaeus chinensis)

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Journal of Ocean University of China 2017年第5期16卷 863-872页

作者： MENG Xianhong SHI Xiaoli KONG Jie LUAN Sheng LUO Kun CAO Baoxiang LIU Ning LU Xia LI Xupeng DENG Kangyu CAO Jiawang ZHANG Yingxue ZHANG Hengheng Key Laboratory for Sustainable Utilization of marine fisheries Resources of 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

To elucidate the molecular response of shrimp hepatopancreas to white spot syndrome virus(WSSV) infection, microarray was applied to investigate the differentially expressed genes in the hepatopancreas of ‘Huanghai No. 2'(Fenneropenaeus chinensis). A total of 59137 unigenes were designed onto a custom-made 60 K Agilent chip. After infection, the gene expression profiles in the hepatopancreas of the shrimp with a lower viral load at early(48–96 h), peak(168–192 h) and late(264–288 h) infection phases were analyzed. Of 18704 differentially expressed genes, 6412 were annotated. In total, 5453 differentially expressed genes(1916 annotated) expressed at all three phases, and most of the annotated were either up-or down-regulated continuously. These genes function diversely in, for example, immune response, cytoskeletal system, signal transduction, stress resistance, protein synthesis and processing, metabolism among others. Some of the immune-related genes, including antilipopolysaccharide factor, Kazal-type proteinase inhibitor, C-type lectin and serine protease encoding genes, were up-regulated after WSSV infection. These genes have been reported to be involved in the anti-WSSV responses. The expression of genes related to the cytoskeletal system, including β-actin and myosin but without tubulin genes, were down-regulated after WSSV infection. Astakine was found for the first time in the WSSV-infected F. chinensis. To further confirm the expression of differentially expressed genes, quantitative real-time PCR was performed to test the expression of eight randomly selected genes and verified the reliability and accuracy of the microarray expression analysis. The data will provide valuable information to understanding the immune mechanism of shrimp's response to WSSV.

关键词： WSSV shrimp Spot differentially Huanghai No.2 Shrimp Infection proteinase custom tubulin

Heterosis and heritability estimates for the survival of the Pacific white shrimp(Litopenaeus vannamei) under the commercial scale ponds

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Acta Oceanologica Sinica 2017年第2期36卷 62-68页

作者： LU Xia LUAN Sheng CAO Baoxiang SUI Juan DAI Ping MENG Xianhong LUO Kun KONG Jie Key Laboratory of Sustainable Utilization of marine fisheries Resources of 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

The aim of the present study is to detect the potential of the base population from diallel crosses of eight introduced strains of the Pacific white shrimp（Litopenaeus vannamei） for improving the yield. Heterosis and heritability were estimated for pond survival at commercial farm conditions for the base population that included 207 full-sib families from a nested mating design by artificial insemination. Among all the hybrids,the heterosis ranged from –11.37%（UA1×UA2） to 20.53%（UA3×SIN） with an average of 0.953%. The results showed that more than half of the hybrids（51.85%） have negative heterosis for survival rate, but most of the hybrids with positive heterosis have high estimates. The high proportion of negative heterosis for survival rate reminders us that the survival trait also should be considered in the crossbreeding program to avoid yield decrease. However, high positive heterosis manifested in most of the hybrids for survival indicates the usefulness of these hybrids for improving the survival to obtain higher yield by crossbreeding in this breeding program. The heritability estimate for pond survival was 0.092±0.043 when genetic groups were included in the pedigree, and it was significantly different from zero（P〈0.05）. The results from this study also indicated that significant improvement for survival is possible through selection in L. vannamei.

关键词： heterosis heritability genetic group pond survival Pacific white shrimp Litopenaeus vannamei

Genetic parameters for cold tolerance and body weight of Chinese fleshy prawn,Fenneropenaeus chinensis

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Acta Oceanologica Sinica 2016年第6期35卷 12-18页

作者： LI Wenjia LUAN Sheng LUO Kun SUI Juan LU Xia WANG Qingyin KONG Jie Key Laboratory for Sustainable Utilization of marine fisheries Resources of Ministry of Agriculture Yellow Sea Fisheries Research InstituteChinese Academy of Fishery Sciences Laboratory for marine fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and Technology

The inability of Fenneropenaeus chinensis to tolerate low temperatures is of major economic concern in temperate climates,as it reduces their growing season and leads to over-winter *** this study,the heritability of body weight under low grow-out temperature and cold tolerance in *** were first investigated and estimated using 88 ful-sib families,which might provide crucial information in Chinese fleshy prawn breeding *** heritability for body weight under suitable and low temperature of *** were both moderate（0.158 0±0.307 5 and 0.132 0±0.026 9 respectively）;the large coefficient of variation（approximately 21%） and moderate estimate of heritability for body weight indicated substantial potential for selective *** heritability estimate for cold tolerance was low（0.019 2±0.023 5）,and showed no significant differences from zero（P〉0.05）.A weak genetic correlation between cold tolerance and body weight was also estimated in the present study,also showing no significant differences from zero（P〉0.05）.Thus,more research needs to be conducted on the more accurate heritability estimate of cold tolerance and genetic correlations between traits in *** to further improve the achievement of breeding goals.

关键词： Fenneropenaeus chinensis heritability body weight cold tolerance genetic correlation

Androgen signaling inhibits de novo lipogenesis to alleviate lipid deposition in zebrafish

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Zoological Research 2024年第2期45卷 355-366页

作者： Jing-Yi Jia Guang-Hui Chen Ting-Ting Shu Qi-Yong Lou Xia Jin Jiang-Yan He Wu-Han Xiao Gang Zhai Zhan Yin College of fisheries Huazhong Agriculture UniversityWuhanHubei 430070China State Key Laboratory of Freshwater Ecology and Biotechnology Institute of HydrobiologyChinese Academy of SciencesWuhanHubei 430072China College of Advanced Agricultural Sciences University of Chinese Academy of SciencesBeijing 100049China Hubei Key Laboratory of Three Gorges Project for Conservation of Fishes Chinese Sturgeon Research InstituteChina Three Gorges CorporationYichangHubei 443100China Hubei Hongshan Laboratory Huazhong Agriculture UniversityWuhanHubei 430070China Innovative Academy of Seed Design Chinese Academy of SciencesBeijing 100049China

Testosterone is closely associated with lipid metabolism and known to affect body fat composition and muscle mass in ***,the mechanisms by which testosterone acts on lipid metabolism are not yet fully understood,especially in *** this study,cyp17a1-/-zebrafish(Danio rerio)exhibited excessive visceral adipose tissue(VAT),lipid content,and up-regulated expression and activity of hepatic de novo lipogenesis(DNL)*** assay for transposase accessible chromatinwithsequencing(ATAC-seq)results demonstrated that chromatin accessibility of DNL genes was increased in cyp17a1-/-fish compared to cyp17a1+/+male fish,including stearoyl-CoA desaturase(scd)and fatty acid synthase(fasn).Androgen response element(ARE)motifs in the androgen signaling pathway were significantly enriched in cyp17a1+/+male fish but not in cyp17a1-/-*** androgen receptor(ar)-/-and wildtype(WT)zebrafish administered with Ar antagonist flutamide displayed excessive visceral adipose tissue,lipid content,and up-regulated expression and activity of hepatic de novo lipogenesis *** Ar agonist BMS-564929 reduced the content of VAT and lipid content,and down-regulated acetyl-CoA carboxylase a(acaca),fasn,and scd ***,the rescue effect of testosterone on cyp17a1-/-fish in terms of phenotypes was abolished when ar was additionally ***,these findings reveal that testosterone inhibits lipid deposition by down-regulating DNL genes via Ar in zebrafish,thus expanding our understanding of the relationship between testosterone and lipid metabolism in teleosts.

关键词： Cyp17a1 Testosterone Androgen receptor De novo lipogenesis Fatty acid synthesis

Comparison of White Spot Syndrome Virus Infection Resistance Between Exopalaemon carinicauda and Litopenaeus vannamei Under Different Salinity Stresses

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Journal of Ocean University of China 2017年第6期16卷 1195-1205页

作者： GE Qianqian YU Ge SUN Ming LI Jitao LI Jian Key Laboratory of Sustainable Development of marine fisheries Ministry of AgricultureYellow Sea Fisheries Research InstituteChinese Academy of Fishery Sciences Function Laboratory for marine fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and Technology

Exopalaemon carinicauda is one of the important economic shrimp species in China, and can tolerate a wide range of salinities. However, its disease resistance remains to be unclear in comparison with other shrimp species under salinity stress. In this study, the resistance to white spot syndrome virus(WSSV) of E. carinicauda and Litopenaeus vannamei was determined by comparing their hemocyanin(Hc) and phenoloxidase(PO) activities under different salinity stresses. In E. carinicauda, the PO activity and Hc gene transcript abundance showed a coherent pattern of increase and decrease while Hc content showed a slightly decrease with Vibrio anguillarum and WSSV infections. For both E. carinicauda and L. vannamei under salinity stress, the PO activity showed a positive correlation with the salinity while the Hc content and expression level of its gene increased significantly in salinities of 5, 15 and 25 g L^(-1). The survival rate of E. carinicauda with WSSV infection was higher than that of L. vannamei in the first 24 h under different salinity stresses. Drastic mortality of *** and L. vannamei appeared at 48 h and 3 h post-injection, respectively. Furthermore, compared with L. vannamei, E. carinicauda displayed higher PO activity, Hc content and abundance of Hc gene m RNA. The results collectively indicated that Hc and PO have obviously functional connection in resisting pathogens and tolerating salinity stress, and PO activity and Hc gene m RNA abundance may reflect the resistance of shrimp to disease. E. carinicauda has higher level of immune potential than L. vannamei, suggesting its greater capacity in resisting pathogens under salinity stresses.

关键词： Exopalaemon carinicauda Litopenaeus vannamei hemocyanin phenoloxidase activity WSSV salinity stress

Effect of Different Dietary Protein and Lipid Levels on the Growth,Body Composition,and Intestinal Digestive Enzyme Activities of Juvenile Yellow Drum Nibea albiflora(Richardson)

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Journal of Ocean University of China 2018年第5期17卷 1261-1267页

作者： WANG Ligai HU Shenyu LOU Bao CHEN Dongxing ZHAN Wei CHEN Ruiyi LIU Feng XU Dongdong Zhejiang marine fisheries Research institute Zhejiang Provincial Key Laboratory of Mariculture & Enhancement marine and Fishery Research institute Zhejiang Ocean University

An 8-week feeding trial was conducted to determine the optimal dietary protein-to-lipid ratio for juvenile Nibea albiflora with an initial weight of(11.76 ± 0.20) *** experimental diets containing different concentrations of protein(40%,47%,or 54%) and lipids(5%,9%,or 13%) in a 3 × 3 factorial experimental design were tested in triplicate groups of fish,while the protein-to-energy(P/E) ratios of the diets varied in the range of 19.74–28.32 mg k J^(-1).Results showed that fish fed diets containing 9% or 13% lipids with 54% protein exhibited significantly higher weight gains and specific growth rates than those fed other *** feed conversion rate of fish fed the diet with 40% protein and 5% lipids was significantly poorer than that of fish fed other *** protein efficiency rate of fish fed diets with 5% lipids was significantly lower than that of fish fed 9% or 13% lipid *** lipid and energy contents were positively correlated with dietary lipid level regardless of protein *** fed a 54% protein diet showed the highest trypsin *** intestinal lipase activity of fish fed the diet containing 13% lipids was significantly higher than that of fish fed 5% or 9% lipid *** results demonstrate the high protein dietary requirements of ***.A diet containing 54% protein and 9%–13% lipids with a P/E ratio of 26.2–27.81 mg protein k J^(-1) can be considered optimal for juvenile ***.

关键词： Nibea albiflora protein to lipid ratio growth performance feed utilization digestive enzyme activities

Lagged response of phytoplanktonic photosynthetic competence to variation of nitrogen concentration in Jiaozhou Bay,China

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Chinese Journal of Oceanology and Limnology 2013年第6期31卷 1295-1305页

作者：付翔李凡吕振波翟红昌韩博平 Shandong Provincial Key Laboratory of Restoration for marine Ecology Shandong Marine Fisheries Research Institute Second institute of Oceanography SOA institute of Hydrobiology Jinan University

Fluorescence-based maximal photochemical efficiency, Fv/Fm, is widely used as an indicator to photosynthetic competence in marine systems. It has been considered a useful parameter diagnosing the nutrient stress on phytoplankton photosynthesis, but many studies argue its usefulness. In the present study, we try to find a temporal relationship between Fv/Fm and nitrogen concentration, and provide a possible explanation on the controversy. We continuously measured Fv/Fm and nitrogen concentration once every 10 days from September 2003 to March 2004 at two stations in Jiaozhou Bay, northern China. It was found that Fv/ffm did not significantly correlate to synchronous nitrogen concentration, but the variation （i.e. the change between two adjacent cruises） of nitrogen concentration of the previous cruise and the variation ofFv/Fm of the current cruise were strongly correlated. This result indicates that a time lag exists between the variation of nutrient status and the subsequent Fv/Fm response. Length of the time lag seems just matched the interval of our measurements （10 days）. In the field, direct dependence of Fv/Fm on nitrogen concentration may not be found because of the lagged response of Fv/Fm to nitrogen concentration variations or physiological acclimation. Our results provide a possible way to explain the previously reported conflicting results on the relationship between Fv/Fm and nutrient status. To give a more-accurate estimate about the length of the time lag, an investigation that includes more frequent measurements is needed.

关键词： maximal photochemical efficiency nitrogen correlation analysis time lag phytoplankton

Distribution and pollution assessment of trace metals in seawater and sediment in Laizhou Bay

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Chinese Journal of Oceanology and Limnology 2015年第4期33卷 1053-1061页

作者：吕冬伟郑冰房燕申格刘惠军 Yantai Research institute ChinaAgricultural University Key Laboratory of marine Ecological Restoration Shandong Marine Fisheries Research Institute National Oceanographic Center

Selected trace metals （Pb, Cd, Cu, Hg） and arsenic in seawater and surface sediments of Laizhou Bay were determined, to evaluate their spatial distribution, pollution risk and potential ecological risk. Concentrations of the elements were 0.56-2.07, 0.14-0.38, 12.70-18.40, 0.014-0.094, and 1.13- 2.37 μg/L in the seawater and 8.94-32.2, 0.18-0.67, 4.51-30.5, 0.006-0.058, and 5.75-15.3 mg/kg in sediments for Pb, Cd, Cu, Hg and As, respectively. High concentrations of the trace metals and arsenic in seawater and surface sediments were generally observed near the fiver estuary. The pollution risk result of the elements showed that Cu was the prominent trace metal pollutant in seawater, followed by Hg, Pb, Cd and As. The metal complex pollution index in seawater was at a medium level. The most important trace metal pollutant in sediments was Cd, followed by As, Cu, Pb, and Hg. Our pollution assessment suggests that trace metal pollution in Laizhou Bay sediments was at a low level. The potential ecological risk was also low in surface sediment.

关键词： Laizhou Bay trace metals arsenic sediments seawater