冻熟虾中副溶血性弧菌生长模型

为控制冻熟虾中副溶血性弧菌的危害,预测冻熟虾中副溶血性弧菌生长情况,对冻熟虾中副溶血性弧菌的生长规律进行了研究。使用CurveExpert 1.3软件作为辅助工具,选取了4种模型(Logistic模型、Richards模型、Compertz模型、MMF模型)拟合了副溶血性弧菌在冻熟虾中生长曲线。结果表明:10℃、18℃和30℃下副溶血性弧菌的生长曲线呈典型的S型,通过标准差和相关系数比较,10℃、18℃和30℃下冻熟虾中副溶血性弧菌最适的生长模型为Richards模型。     

副溶血弧菌噬菌体微胶囊的制备及在饵料中的应用

采用复相乳液法,以乙基纤维素为包囊材料,明胶溶液为保护液制取微胶囊,对副溶血弧菌噬菌体(Vibrio parahaemolyticus phage)进行固定化,制备噬菌体微胶囊。结果显示,乙基纤维素质量浓度为30 g/L,明胶质量浓度为20 g/L,搅拌速度为600 r/min,时间30 min,副溶血弧菌噬菌体悬液与乙基纤维素溶液的体积比为1∶3时,所得微胶囊有较好的噬菌体缓释性能,综合评价指数最高。对方斑东风螺(Babylonia areolata)饲以混有噬菌体微胶囊的饵料,当饵料中噬菌体微胶囊质量分数为1.0 g/kg时,与对照组比较,东风螺肠道宿主菌平均减少96%,水体中宿主菌减少72%,差异均有统计学意义(P0.05);以黏附剂按1.5 g/kg的剂量将噬菌体微胶囊黏附于对虾饵料,并投喂凡纳滨对虾(Litopenaeus vannamei),对虾肠道的宿主菌比对照组平均减少86%,水体中宿主菌比对照组减少68%,差异均有统计学意义(P0.05)。微胶囊可有效控制宿主弧菌的在养殖体系的浓度,减少养殖动物弧菌病的产生。     

Biodeposition,respiration,and excretion rates of an introduced clam Mercenaria mercenaria in ponds with implications for potential competition with the native clam Meretrix meretrix in Shuangtaizi estuary,China

This study aimed to evaluate the potential impacts of an introduced clam Mercenaria mercenaria on estuarine ecosystem,and implications for the niche competition with a native clam M eretrix meretrix.The biodeposition,respiration,and excretion rates of M.mercenaria were determined seasonally using a sediment trap and a closed respirator in field.The biodeposition rates of M.mercenaria were 0.06–0.37 g/(ind.·d),and the respiration rates were 0.31–14.66 mg/(ind.·d).The ammonia and phosphate excretion rates were 0.18–36.70 and 1.44–14.87 μg/(ind.·d),respectively.The hard clam M.mercenaria may discharge dry deposits up to 2.1×10~5 t,contribute 18.3 t ammonia and 9.0 t phosphate to culture ponds,and consume 7.9×10~3 t O_2 from ponds annually.It suggested that the hard clam M.mercenaria might play an important role in pelagic-benthic coupling in pond ecosystem through biodeposition and excretion.A comparison of the key physiological parameters of the introduced clam M.mercenaria and the native clam Meretrix meretrix suggested that M.mercenaria had a niche similar to that of Meretrix meretrix in Shuangtaizi estuary and might have a potential competition with Meretrix meretrix for habitat and food if M.mercenaria species escaped from the culture pond or artificially released in estuarine ecosystem.     

Immune response of Pseudosciaena crocea to the injection of Vibrio alginolyticus

For the investigation of anti-infection immune response of Pseudosciaena crocea, 160 healthy fish samples were categorized into infected and control groups. Each individual fish in the infected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension of Vibrio alginolyticus in density of 2×107 CFU/ml, while each individual in the control group was injected i.p. with 0.2 ml sterile saline solution (0.85%). It was observed that the artificial injection of V. alginolyticus significantly inc...     

无菌人工养殖海水中副溶血弧菌噬菌体与宿主的消长动态

在3组一定浓度的副溶血弧菌菌液中分别加入3种浓度的副溶血弧菌噬菌体,第12天各实验组再次分别加入一定浓度的副溶血弧菌或副溶血弧菌噬菌体,比较各组水体中副溶血弧菌噬菌体及其宿主的动态关系。结果表明,副溶血弧菌噬菌体可使宿主下降至一定浓度并与之保持动态平衡关系,实验组1、2、3中副溶血弧菌浓度从2.43×106 mL-1分别下降至6.2×104、9.2×104、7.46×105 mL-1,并以此浓度范围维持下去。高浓度副溶血弧菌噬菌体与其宿主之间平衡关系的稳定性较好,再次加入副溶血弧菌,高、中浓度组分别在1 d和4 d后副溶血弧菌下降至平衡时的数量级浓度,并持续下去。低浓度组副溶血弧菌组有所上升,并持续下去。再次加入副溶血弧菌噬菌体对副溶血弧菌浓度影响较小(P>0.05)。     

The influence of salinity on toxicological effects of arsenic in digestive gland of clam Ruditapes philippinarum using metabolomics

Ruditapes philippinarum,a clam that thrives in intertidal zones of various salinities,is a useful biomonitor to marine contaminants.We investigated the influence of dilution to 75% and 50% of normal seawater salinity(31.1) on the responses of the digestive gland of R.philippinarum to arsenic exposure(20 μg/L),using nuclear magnetic resonance(NMR)-based metabolomics.After acute arsenic exposure for 48 h,salinity-dependent differential metabolic responses were detected.In normal seawater,arsenic exposure increased the concentrations of branched-chain amino acids,and of threonine,proline,phosphocholine and adenosine,and it decreased the levels of alanine,hypotaurine,glucose,glycogen and ATP in the digestive glands.Differential changes in metabolic biomarkers observed at lower salinity(～23.3) included elevation of succinate,taurine and ATP,and depletion of branched-chain amino acids,threonine and glutamine.Unique effects of arsenic at the lowest salinity(～15.6) included down-regulation of glutamate,succinate and ADP,and up-regulation of phosphocholine.We conclude that salinity influences the metabolic responses of this clam to arsenic.     

Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection

The appropriate reference gene is a prerequisite for accurate normalization of gene expression level, and research on suitable reference genes in clam Cyclina sinensis is scarce. To improve the situation, we selected five commonly used housekeeping genes, including β-actin, Elongation factor 1-α (EF1-α), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 40S ribosomal protein S18 (RPS18), and Tubulin α (TUB-α), then evaluated their expression stability in different adult tissues and under different experimental treatments (salinity stress and Vibrio parahaemolyticus infection). Their expression stability was analyzed by three frequently used programs, geNorm, NormFinder, and BestKeeper. This analysis indicated that multiple genes should be used for normalization, and we concluded that the reference gene combination GAPDH-RPS18-β-actin, should be used for qRT-PCR analysis in different tissues of C. sinensis under normal physiological conditions. For the clams under salinity stress and Vibrio infection, EF1-α-GAPDH-RPS18 was recommended as the gene combination for qRT-PCR normalization. TUB-α was generally poorly ranked by all programs, and should not be used in future studies. This study should provide fundamental support for accurate quantitative gene expression analysis of this species.

     

Effect of cadmium on the defense response of Pacific oyster Crassostrea gigas to Listonella anguillarum challenge

Heavy metal pollution can affect the immune capability of organisms.We evaluated the effect of cadmium(Cd) on the defense responses of the Pacific oyster Crassostrea gigas to Listonella anguillarum challenge.The activities of several important defensive enzymes,including superoxide dismutase(SOD),glutathione peroxidase(GPx),acid phosphatase(ACP),Na+,K+-ATPase in gills and hepatopancreas,and phenoloxidase-like(POL) enzyme in hemolymph were assayed.In addition,the expression levels of several genes,including heat shock protein 90(HSP90),metallothionein(MT),and bactericidal/permeability increasing(BPI) protein were quantified by fluorescent quantitative PCR.The enzyme activities of SOD,ACP,POL,and GPx in hepatopancreas,and the expression of HSP90 were down-regulated,whereas GPx activity in the gill,Na+,K+-ATPase activities in both tissues,and MT expression was increased in Cdexposed oysters post L.anguillarum challenge.However,BPI expression was not significantly altered by co-stress of L.anguillarum infection and cadmium exposure.Our results suggest that cadmium exposure alters the oysters’ immune responses and energy metabolism following vibrio infection.     

Status and historical changes in the fish community in Erhai Lake

Erhai Lake is the second largest freshwater lake on the Yunnan Plateau, Southwest China. In recent decades, a number of exotic fish species have been introduced into the lake and the fish community has changed considerably. We evaluated the status of the fish community based on surveys with multimesh gillnet, trap net, and benthic fyke-net between May 2009 and April 2012. In addition, we evaluated the change in the community using historical data (1952–2010) describing the fish community and fishery harvest. The current fish community is dominated by small-sized fishes, including Pseudorasbora parva, Rhinogobius giurinus, Micropercops swinhonis, Hemiculter leucisculus, and Rhinogobius cliffordpopei. These accounted for 87.7% of the 22 546 total specimens collected. Omnivorous and carnivorous species dominated the community. A canonical correspondence analysis (CCA) plot revealed that the distribution of fishes in the lake is influenced by aquatic plants, water temperature, pH, and season. The abundance of indigenous species has declined sharply, and a majority of endemic species have been extirpated from the lake (a decrease from seven to two species). In contrast, the number of exotic species has increased since the 1960s to a total of 22 at present. The fishery harvest decreased initially following the 1960s, but has since increased due to the introduction of non-native fish and stocking of native fish. The fishery harvest was significantly correlated with total nitrogen, not total phosphorus, during the past 20 years. Based on our results, we discuss recommendations for the restoration and conservation of the fish resources in Erhai Lake.     

Bacterial community succession in response to dissolved organic matter released from live jellyfish

Jellyfish blooms have increased worldwide,and the outbreaks of jellyfish population not only affect the food web structures via voracious predation but also play an important role in the dynamics of nutrients and oxygen in planktonic food webs.However,it remains unclear whether specific carbon compounds released through j ellyfish metabolic processes have the potential to shape bacterial community composition.Therefore,in this study,we aimed to investigate the compositional succession of the bacterioplankton community in response to the dissolved organic matter(DOM)released by the live Scyphomedusae Cyanea lamarcki'i and Chrysaora hysoscella collected from Helgoland Roads of the North Sea.The bacterial community was significantly stimulated by the DOM released form live jellyfish and different dominant phylotypes were observed for these two Scyphomedusae species.Furthermore,the bacterial community structures in the different DOM sources,jellyfish-incubated media,Kabeltonne seawater,and artificial seawater(DOM-free)were significantly different,as revealed by automated ribosomal intergenic spacer analy sis fingerprints.Catalyzed reporter depo sition fluorescence in situ hybridization(CARD-FISH)revealed a rapid species-specific shift in bacterial community composition.Gammaproteobacteria dominated the community instead of the Bacteroidetes community for C.lamarckii,whereas Gammaproteobacteria and Bacteroidetes dominated the community for C.hysoscella.The significant differences in the bacterial community composition and succession indicate that the components of the DOM released by jellyfish might differ with jellyfish species.     

Seasonal changes in the demersal nekton community off the Changjiang River estuary

The diversity, community structure and seasonal variation in demersal nekton off the Changjiang （Yangtze） River estuary was evaluated using monthly trawl survey data, collected between December 2008 and November 2009. A total of 95 species （56 teleosts, 11 cephalopods, and 28 decapod crustaceans） from 69 genera, 49 families and 15 orders were collected. These species could be classified into six groups on the basis of temporal distribution patterns. The resident crab Ovalipes punctatus dominated the community, both in number and biomass. A clear seasonal succession was observed in the species composition. Cluster analysis revealed three primary seasonal groups corresponding to the samples collected in winter-spring, late spring-summer and late summer-autumn. The highest biomass and lowest diversity were observed in summer, while the lowest biomass and highest diversity in winter. The abundance-biomass comparison curves and community composition suggested that the investigated community was moderately disturbed. The results suggest that reduction in fishing pressure and in the degree of seasonal hypoxia are essential for sustainable resource management off the Changjiang River estuary.     

Ontogenetic changes in digestive enzyme activities and the amino acid profile of starry flounder Platichthys stellatus

Ontogenetic changes in digestive enzyme activities and the amino acid(AA) profile of starry flounder,Platichthys stellatus,were investigated and limiting amino acids were estimated compared with the essential AA profile between larvae and live food to clarify starry flounder larval nutritional requirements.Larvae were collected at the egg stage and 0,2,4,7,12,17,24 days after hatching(DAH)for analysis.Larvae grew from 1.91 mm at hatching to 12.13 mm at 24 DAH.Trypsin and chymotrypsin activities changed slightly by 4 DAH and then increased significantly 4 DAH.Pepsin activity increased sharply beginning 17 DAH.Lipase activity increased significantly 4 DAH and increased progressively with larval growth.Amylase activity was also detected in newly hatched larvae and increased 7 DAH followed by a gradual decrease.High free amino acid(FAA) content was detected in starry flounder eggs(110.72mg/g dry weight).Total FAA content dropped to 43.29 mg/g in 4-DAH larvae and then decreased gradually to 13.74 mg/g in 24-DAH larvae.Most FAAs(except lysine and methionine) decreased 50%in 4-DAH larvae compared with those in eggs and then decreased to the lowest values in 24-DAH larvae.Changes in the protein amino acid(PAA) profile were much milder than those observed for FAAs.Most PAAs increased gradually during larval development,except lysine and phenylalanine.The percentages of free threonine,valine,isoleucine,and leucine decreased until the end of the trial,whereas the protein forms of these four AAs followed the opposite trend.A comparison of the essential AA composition of live food(rotifers,Artemia nauplii,and Artemia metanauplii) and larvae suggested that methionine was potentially the first limiting AA.These results may help develop starry flounder larviculture methods by solving the AA imbalance in live food.Moreover,the increased digestive enzyme activities indicate the possibility of introducing artificial compound feed.     

创伤弧菌和哈氏弧菌双重PCR检测方法的建立及初步应用

根据创伤弧菌(Vibrio vulnificus)的溶细胞毒素基因序列和哈氏弧菌(Vibrio harveyi)的toxR基因序列,分别设计并合成两对特异性引物,通过PCR反应条件优化,测试两种菌的特异性和敏感性,建立双重PCR方法,同时快速检测V.vulnificus和V.harveyi。结果表明:纯培养V.vulnificus和V.harveyi的检测灵敏度分别是12 cfu/mL和18 cfu/mL,与无乳链球菌、海豚链球菌、副溶血弧菌及美人发光杆菌无交叉反应;此PCR检测方法具有良好的特异性、敏感性,具快速、高效等优点,对细菌V.vulnificus和V.harveyi诊断与防治具有较好的临床应用性。     

Spatio-temporal distribution of net-collected phytoplankton community and its response to marine exploitation in Xiangshan Bay

To explore the spatial-temporal distribution of the phytoplankton community and evaluate the combined effects of marine resource exploitation, net-collected phytoplankton and physical-chemical parameters were investigated in the Xiangshan Bay during the four seasons of 2010. A total of eight phyla, 97 genera, and 310 species were found, including 232 diatom species, 45 dinoflagellate species and 33 other taxa. The phytoplankton abundances presented a significant (P<0.001) seasonal difference with the average of 60.66×10⁴ cells/m³. Diatoms (mainly consisting of Coscinodiscus jonesianus, Cerataulina pelagica, Skeleto n ema costatum, and genus Chaetoceros) dominated the phytoplankton assemblage in all seasons. We found great spatio-temporal variation in community composition based on the multidimensional scaling and similarity analysis. Canonical correspondence analysis show that temperature, nutrition, illumination, and salinity were the main variables associated with microalgal assemblage. Compared with the previous studies, an increase in phytoplankton abundance and change in the dominant species coincided with increased exploitation activities in this bay (e.g. operation of coastal power plants, intensive mariculture, tidal flat reclamation, and industrial and agricultural development). The present findings suggest that the government should exercise caution when deciding upon developmental patterns in the sea-related economy.     

Response of rotifer community to environmental changes in five shallow lakes in the middle reach of Changjiang River,China

We evaluated the response of a rotifer community to environmental changes in five shallow lakes in the middle reach of the Changjiang （Yangtze） River in summer and autumn 2010. These five lakes differed in trophic status and rotifer community structure. Twenty-nine rotifer species were collected, of which Keratella cochlearis, Polyarthra dolichoptera, and Trichocerca elongate were dominant. The mean density, Shannon-Wiener diversity, and equitability among the five lakes differed significantly （P〈0.05）. The mean rotifer density was positively correlated with trophic state. The diversity was higher in lakes with high macrophyte coverage. The composition of rotifer species was closely associated with the trophic gradient. Five environmental variables, Secchi depth, conductivity, TN, NHa-N, and TP, significantly affected the composition of rotifer species. Keratella quadrata, Brachionus calyciflorus, and B. forficula were more common in eutrophic conditions. Our results suggest that eutrophication has a significant influence on the rotifer community structure and highlight the potential for using rotifer community structure as an indicator of trophic status in subtropical lakes.     

Quantitative proteomics provides insight into the response of the marine dinoflagellate Prorocentrum donghaiense to changes in ambient phosphorus

Journal of Oceanology and Limnology - Dinoflagellates are the major causative agents of harmful algal blooms in the global ocean and they usually form blooms under conditions of very low dissolved...     

Effects of temperature, pH and NaC1 on protease activity in digestive tract of young turbot, Scophthalmus maximus

The protease activity in digestive tract of young turbot Scophthalmus maximum was studied, and the optimal pH, temperature and NaCl concentration were determined for different portions of the fish＇s internal organs. The optimal activity in the fish＇s stomach was at pH of 2.2, while that in the intestinal extracts was within the alkaline range from 9.5 to 10.0. In hepatopancreas, the optimal pH was in low alkalinity at 8.5. The optimal reaction temperature was above 40℃ in stomach, intestine and hepatopancreas. With increasing temperature, the pH value increased in stomach, while in the intestine, an opposite tendency was observed due to combined effect of pH and temperature. NaCl concentration showed inhibitory impact on protein digestion in hepatopancreas. The main protease for protein digestion in turbot seemed to be pepsin. Moreover, the maximum protease activity in different segments of intestine existed in the hindgut.     

Effects of temperature, pH and NaCl on protease activity in digestive tract of young turbot, Scophthalmus maximus

1 INTRODUCTION Digestive enzyme activity is one of important issues for learning digestive physiology of fish, and widely applied in commercial fish culture. So far, the data obtained in fish showed that the diges- tive enzymes were qualitatively similar …     

Molecular characterization and expression analysis of Lily-type lectin (SmLTL) in turbot Scophthalmus maximus,and its response to Vibrio anguillarum

A full-length lily-type lectin(Sm LTL) was identified from turbot(Scophthalmus maximus) in this study. By searching database for protein identification and function prediction, Sm LTL were confirmed. The full-length c DNA of Sm LTL is composed of 569 bp and contains a 339 bp ORF that encodes 112 amino acid residues. The Sm LTL peptide is characterized by a specific β-prism architecture and contains three mannose binding sites in a three-fold internal repeat between amino acids 30–99; two of the repeats share the classical mannose binding domain(Qx Dx Nx Vx Y) while the third binding site was similar to other fish-specific binding motifs(Tx Tx Gx Rx V). The primary, secondary, and tertiary structures of Sm LTL were predicted and analyzed, indicating that the S m LTL protein was hydrophilic, contained 5.36% α-helices, 39.29% extended strands, 16.07% β-folds, and 39.29% random coils, and three β-folds. Quantitative realtime polymerase chain reaction(qPCR) analysis revealed that the Sm LTL mRNA was abundantly expressed in skin, gill, and intestine. Low levels of Sm LTL expression were observed in other tissues. The expression of Sm LTL in gill, skin and intestine increased at m RNA level after stimulation of V ibrio anguillarum, our results suggest that Sm LTL serve as the first line of defence against microbial infections and play a pivotal role in the innate mucosal immune system. The current study indicates that Sm LTL is a member of the lilytype lectin family and the information reported here will provide an important foundation for future research on the role of this protein.