Effects of temperature and salinity on survival,growth and DNA methylation of juvenile Pacific abalone,Haliotis discus hannai Ino

Temperature and salinity are two of the most potent abiotic factors infl uencing marine mollusks.In this study,we investigated the individual and combined ef fects of temperature and salinity on the survival and growth of juvenile Pacifi c abalone,Haliotis discus hannai Ino,and also examined the DNA methylation alteration that may underpin the phenotypic variation of abalone exposed to dif ferent rearing conditions.The single-factor data showed that the suitable ranges of temperature and salinity were 16–28°C at a constant salinity of 32,and 24–40 at a constant temperature of 20°C,respectively.The two-factor data indicated that both survival and growth were signifi cantly af fected by temperature,salinity and their interaction.The optimal temperature-salinity combination for juveniles was 23–25°C and 30–36.To explore environment-induced DNA methylation alteration,the methylation-sensitive amplifi ed polymorphism(MSAP)technique was used to analyze the genomic methylation profi les of abalone reared in optimal and adverse conditions.Neither temperature nor salinity induced evident changes in the global methylation level,but 67 and 63 dif ferentially methylated loci were identifi ed in temperature and salinity treatments,respectively.The between-group eigen analysis also showed that both temperature and salinity could induce epigenetic dif ferentiation in H.discus hannai Ino.The results of our study provide optimal rearing conditions for juvenile H.discus hannai Ino,and represent the fi rst step toward revealing the epigenetic regulatory mechanism of abalone in response to thermal and salt stresses.     

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.

     

Salinity tolerance and genetic diversity of the dinoflagellate <Emphasis Type="Italic">Oxyrrhis marina</Emphasis>

To evaluate the relationship between salinity tolerance and genetic diversity of plankton, we collected a wild species of plankton from Taipingjiao, Qingdao. The fragment of ITS1-5.8S rDNA-ITS2 was extracted and sequenced. The results showed that the plankton belongs to Oxyrrhis marina. The salinity tolerance of O. marina ranges from 4 to 60. Seven selected groups were built up to evaluate salinity tolerance and to assess genetic diversity by RAPD. The salinity tolerance comparison revealed considerable differences among groups: the strains of O. marina in group 4 could survive under salinity from 4 to 32, while the strains selected for salinity 60 died under the salinity lower than 16. Analysis of genetic diversity of the seven groups showed that the mean genetic diversity index value was 0.28, but it was only 0.16 in selected group of 4 and was 0.24 for group 60. The result of AMOVA suggested a significantly positive relationship between the salinity tolerance and genetic diversity of O. marina (P<0.01). This study indicates that consideration of intraspecific genetic divergence in O. marina might be indispensable when using it as a model in the study of salinity tolerance of wild plankton.     

FOOD SOURCES AND CARBON BUDGET OF CHINESE PRAWN PENAEUS CHINENSIS

This study deals with contribution of artificial food pellet and natural food to Chinese prawn (Penaeus orientalis) growth in a semiintensive culture pond. The prawn carbon consumption, budget, and the effects of some factors on the budget were investigated. The results showed that 26.2% of P. orientalis growth carbon came from formulated feed at the initial culture stage (when the prawns were 0.06±0.01 g in wet weight), and was 62.5% when the prawns were 9.56±1.04 g. The remaining part of the growth carbon was derived from organic fertilizer and natural food. The highest growth rate occurred at 20×10-3 salinity. Suitable salinity for culturing Chinese prawn was (20-28)×10-3.     

Effects of salinity fluctuation frequency on the growth, molting rate and hemolymph 20-hydroxyecdysone concentration in Juvenile Chinese shrimp, Fenneropenaeus chinensis

This experiment was conducted to examine the effects of salinity fluctuation frequency on the growth, molting rate and hemolymph 20-hydroxyecdysone (20-HE) concentration for juvenile Fenneropenaeus chinensis with an initial wet body weight of 1.115 g±0.012 g. The salinity of the control treatment (represented by D0) was 28 throughout the experiment, whereas treatments D2, D4, D6 and D8 were subjected to different salinity fluctuation frequencies of 2, 4, 6 and 8 d, respectively. The range of salinity fluctuation was 4. The salinity in treatments D2, D4, D6 and D8 was kept at 28 for 2, 4, 6 and 8 d, respectively; each original salinity was decreased abruptly to salinity 24, which first lasted for another 2 d and was then raised to its initial value 28. This constituted a salinity fluctuation cycle and afterwards the cycle repeated. It was found that: 1) The weight gain of shrimp in treatment D4 was 202.9% and significantly higher than that in treatment D0 (P<0.05). The molting rate in treatment D2 was the lowest, while that in treatment D6 was the highest and significantly higher than those in treatments D0 and D2 (P<0.05). 2) The hemolymph 20-HE concentration of shrimps in all treatments was at low levels and increased first gradually during the inter-molt period and then increased sharply. It reached a peak value during the pre-molt stage, and then decreased abruptly during the post-molt stage when the lowest level occurred. 3) During the post-molt stage, the hemolymph 20-HE concentration in treatment D2 was 7.47 pg μL⁻¹ and significantly higher than that in treatment D0 (P<0.05). There were, however, no significant differences in hemolymph 20-HE concentration among all five treatments during the inter-molt stage (P>0.05). During the pre-molt stage, the hemolymph 20-HE concentration in treatment D6 was significantly higher than that in treatment D0 (P<0.05), whereas the hemolymph 20-HE concentration in treatment D2 was significantly lower than that in other treatments (P<0.05). Adequate salinity fluctuation promoted the molting rate through increasing hemolymph 20-HE concentration at the pre-molt stage.     

温度和盐度对吉富罗非鱼幼鱼肠道两种抗氧化酶活力的联合效应

采用中心组合设计和响应曲面法研究温度(12~34℃)和盐度(0~26)对吉富品系尼罗罗非鱼(Oreochromis niloticus)幼鱼(体长4.20±0.10 cm,体质量2.42±0.15 g)肠道超氧化物歧化酶(Superoxide Dismutase,SOD)、过氧化氢酶(Catalane,CAT)的联合效应。结果表明:1)温度对吉富罗非鱼幼鱼肠道SOD和CAT活力的线性效应和累积效应均极显著(P<0.01);盐度对吉富罗非鱼幼鱼肠道SOD活力线性效应显著(P<0.05),累积效应极显著(P<0.01);盐度对吉富罗非鱼幼鱼肠道CAT活力线性效应不显著(P>0.05),累积效应极显著(P<0.01);温度和盐度对SOD活力的互作效应极显著(P<0.01),对CAT活力的互作效应不显著(P>0.05)。2)响应曲面法分析表明,随着温度和盐度的增大,两种抗氧化酶的活力均呈先减小后增大的趋势。3)建立两种抗氧化酶活力与温度、盐度间关系的模型方程(SOD:决定系数R2=0.992 9,预测决定系数R2Pred.=0.939 2,P<0.01;CAT:决定系数R2=0.948 5,预测决定系数R2Pred.=0.647 1,P<0.01),两模型方程可用于预测吉富罗非鱼幼鱼肠道SOD和CAT的活力。4)优化结果显示,温度为24.15℃,盐度为11.75时,SOD活力最小值为9.80 U/mg,CAT活力最小值为9.28 U/mg,满意度函数值高达0.961。     

CHARACTERIZATION OF LACTATE DEHYDROGENASE ISOZYME PATTERN AND MORPHOLOGY OF THREE MARINE FISH CELL LINES

Three continuous marine fish cell lines of FG (i. e., Hounder Gill) from flounder (Paralichthys olivaceus) gill, SPH (i. e. , Sea Perch Heart) from sea perch (Lateolabrax japonicus) heart and RSBF (i.e., Red Sea Bream Fin) from red sea bream (Pagrosomus major) fin, were characterized by lactate dehydrogenase (LDH) isozyme and morphological analysis. The LDH isozyme patterns of these three cell lines and their corresponding tissues of origin were investigated and compared. The results showed: (1) No difference was found in the LDH isozyme patterns of FG and flounder gill tissue. However, the LDH isozyme patterns of SPH and RSBF were significantly different from their corresponding tissues of origin; (2) LDH isozyme patterns of FG, SPH and RSBF were markedly different from each other and could serve as genetic markers for species identification and detection of cross contamination. Morphological change analysis of these three cell lines in comparison to their original tissues indicated that FG cells still appeared epithelioid without morphological transformation. However, morphological changes were found in SPH and RSBF compared to their original tissues. Therefore, the cellular morphology was still plastic in the relatively stable culture conditions, and it was possible that change of LDH patterns wasrelated to morphological changes of fish cells in vitro.     

Ontogenic study of isozymes in giant tiger prawn,Penaeus monodon (Crustacea: Decapoda)

The patterns of isozyme variation throughout the life cycle of the giant tiger prawn,Penaeus monodon were described for ten enzyme systems (G3PD, MDH, LGG, GPI, MPI, FBP, PGM, LDH, PGD, and PROT). The zymograms showed little variation in the number of isozymic loci expressed during larval development compared with results reported for other penaeid prawns. There were specific ontogenic patterns at some loci. As larvae developed beyond the embryonic stage, there was a tendency for intensity of staining and complexity of banding patterns to increase.     

Variations and controlling factors of the coccolith weight in the Western Pacific Warm Pool over the last 200 ka

Using a coccolith weight analytic software (Particle Analyser), we analyze most abundant coccolith species in a sediment core from the central Western Pacific Warm Pool (WPWP) and calculate coccolith size and weight variations over the last 200 ka. These variations are compared with the trends of sea surface temperature (SST), primary productivity (PP), sea surface salinity (SSS), and insolation. Our results demonstrate that the size and weight of the coccoliths varied in response to variations of these factors, and their average total weight is primarily related to the relative abundance of the dominant species GEO (Gephyrocapsa oceanica). The variation in weight of EMI (Emiliania huxleyi) and GEE (Gephyrocapsa ericsonii) are mainly influenced by nutrients, and the variation of GEM (G. muellerae conformis) and GEO (G. oceanica) weight are mainly influenced by SST. For all of the taxa weight, PP and SST present apparent precession or semi-precession cycles, we consider that the mono-coccolith weight of the Equatorial Western Pacific is primarily affected by precession drived thermocline and nutricline variation.     

Oxygen consumption and ammonia-N excretion of Meretrix meretrix in different temperature and salinity

Effects of temperatures and salinities on oxygen consumption and ammonia-N excretion rate of clamMeretrix meretrix were studied in laboratory from Oct. 2003 to Jan. 2004. Two schemes were designed in incremented temperature at 10, 15, 20, 25°C at 31.5 salinity and in incremented salinity at 16.0, 21.0, 26.0, 31.5, 36.0, and 41.0 at 20°C, all for 8–10 days. From 10 to 25°C, both respiration and excretion rate were increased. One-way ANOVA analysis demonstrated significant difference (P<0.01) in physiological parameters in this temperature range except between 15 and 20°C. The highestQ ₁₀ thermal coefficient value (12.27) was acquired between 10 and 15°C, and about 1 between 15 and 20°C, indicatingM. meretrix could well acclimate to temperature changes in this range. Salinity also had significant effects on respiration and excretion rate (P<0.05). The highest values of respiration and excretion rate ofM. meretrix were recorded at 16.0 salinity (20°C). These two physiological parameters decreased as salinity increased until reached the minimumQ ₁₀ value at 31.5 (20°C), then again, these parameters increased with increasing salinity from 31.5 to 41.0.M. meretrix can catabolize body protein to cope with osmotic pressure stress when environmental salinity is away from its optimal range. No significant difference was observed between 26.0 and 36.0 in salinity (P>0.05), suggesting that a best metabolic salinity range for this species is between 26.0 and 36.0. This work is supported by National High-Tech R & D Program of China. (863 Program) (2002AA603014).     

Influence of salinity and nitrogen content on production of dimethylsulfoniopropionate (DMSP) and dimethylsulfide (DMS) by Skeletonema costatum

The effects of changing salinity and nitrogen limitation on dimethylsulfoniopropionate(DMSP) and dimethylsulfide(DMS) concentrations were investigated in batch cultures of coastal diatom Skeletonema costatum,an ecologically important species.Changes in salinity from 20-32 caused no measurable variation in cell growth or culture yield,but increased intracellular DMSP per cell by 30%.Nitrogen limitation caused up to a two-fold increase in total DMSP per cell and up to a three-fold increase in DMS per cell.These changes in DMSP and DMS per cell in the Skeletonema costatum cultures with nitrogen limitation and changing salinity were primarily attributed to the physiological functions of DMSP as an osmolyte and an antioxidant.The data obtained in this study indicated that nitrogen limitation and salinity may play an important role in climate feedback mechanisms involving biologically derived DMS.     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

Effects of salinity fluctuation frequency on the osmolarity, Na+-K+-ATPase activity and HSP70 expression in juvenile chinese shrimp, Fenneropenaeus chinensis

Experiments were conducted to examine the effects of salinity fluctuation frequency on the osmolarity, Na⁺-K⁺-ATPase activity and HSP70 of Chinese shrimp Fenneropenaeus chinensis with initial wet body weight of 1.460 g ± 0.091 g. The salinity in the control group (D0) was 28 throughout the experiment, whereas treatments D2, D4, D6 and D8 were subjected to different salinity fluctuation frequencies of 2, 4, 6 and 8 d, respectively. The salinity in treatments D2, D4, D6 and D8 was kept at 28 for 2, 4, 6 and 8 d, respectively, decreased abruptly to salinity 24, lasted for another 2 d, and then was raised to its initial value 28. This was a complete salinity fluctuation cycle that afterwards repeated itself. After 32 days, the osmolarity in treatments D2, D4, D6 and D8 was significantly lower than that in treatment D0 (P<0.05). There were significant differences in both muscle and eyestalks HSP70 expression among groups. The HSP70 expressions in muscle and eyestalks in group D4 were 61.4% and 57.0% higher, respectively, than that in the control group D0 (P<0.05). There were, however, no significant differences in gill or hepatopancreas Na⁺-K⁺-ATPase activity between the treatments and the control.     

EFFECTS OF FISH OIL,DHA OIL AND LECITIHIN IN MICROPARTICULATE DIETS ON STRESS TOLERANCE OF LARVAL GILTHEAD SEABREAM（SPARUS AURATA）

The effects of natural fish oil,DHA oil and soybean lecithin in microparticulate diets on stress tolerance of larval gilthead seabream(Sparus aurata)were investigated after 15 days feeding trials.The tolerance of larval gilthead seabream to various stress factors such as exposure to air(lack of dissolved oxygen),changes in water temperature(low)and salinity(high) were determined.This study showed that microparticulate diet with natural fish oil and soybean lecithin was the most effective for in-creasing the tolerance of larval gilthead seabream to various stresses,and that microparticulate diet with natural fish oil and palmitic acid(16:0)was more effective than microparticulate diet with DHA oil and soybean lecithin.     

The occurrence of an exotic bisexual Artemia species, Artemia franciscana , in two coastal salterns of Shandong Province, China

The alien halophilous Artemia species, Artemia franciscana, was found in Chengkou Saltern and Yangkou Saltern of Shandong Province, P.R. China. Although the indigenous parthenogenetic Artemia is detectable, the exotic species is dominant in both salterns. The cross-breeding tests between the exotic A. franciscana and 5 bisexual Artemia species were conducted. The results of hybridization and morphological observations on the exotic A. franciscana are briefly presented in this short communication.     

A calibration method of Argo floats based on multiple regression analysis

1 INTRODUCTION Argo floats are instruments that move freely with the ocean current at fixed parking depths and cycle from a profiling depth to the sea surface at regular time intervals. While rising to the surface, these autonomous floats take profiles of…     

Occurrence and diversity of Candida genus in marine environments

A total of 317 yeast isolates from seawater, sediments, mud of salterns, guts of marine fishes and marine algae were obtained. The results of routine identification and molecular characterization showed that six isolates among these marine yeasts belonged to Candida genus as Candida intermedia for YA01a, Candida parapsilosis for 3eA2, Candida quercitrusa for JHSb, Candia rugosa for wl8, Candida zeylanoides for TJY13a, and Candida membranifaciens for W14-3. Isolates YA01a (Candida intermedia), wl8 (Candida rugosa), 3eA2 (Candida parapsilosis), and JHSb (Candida quercitrusa) were found producing cell-bound lipase, while isolate W14-3 (Candida membranifaciens) producing riboflavin. These marine yeast Candida spp. seem to have wide potential applications in biotechnology.     

近江牡蛎3个种群生化遗传变异的研究

用同工酶电泳方法研究广东省官渡、阳江和汕尾的近江牡蛎(Ostea rivularis)的遗传变异。分析了11种同工酶20个的位点,其中出现13个位点显示为多态。结果表明官渡、阳江、汕尾种群的多态位点比例分别为0.200、0.200、0.500。官渡和阳江种群间,官渡和汕尾种群间以及阳江和汕尾种群间的遗传距离(D)分别为0.0451,0.1125,0.1088。     

Seasonal change in body length of important small copepods and relationship with environmental factors in Jiaozhou Bay,China

Differences among species in prosome length and in species’ response to environmental factors do exist. Therefore, it is useful to examine prosome length for different copepod species in variable environments. Seasonal variations in prosome length of four small copepods and their copepodite stages in the Jiaozhou Bay were compared and the relative influence of temperature, salinity, and chlorophyll concentration were examined. Two peaks were found in the mean prosome length of Paracalanus parvus (in early winter and May). For Acartia bifilosa, the maximum values of all copepodites occurred mainly from February to April, and decreased to the bottom in July. Prosome length of Acartia pacifica peaked when it first appeared in June, then reached to the minimum in July. Parvocalanus crassirostris only appeared from late summer to autumn and the mean prosome length showed no clear changes. Correlations of adult prosome length with environmental factors were evaluated. For the four species, temperature was negatively correlated to prosome length except for P. crassirostris. But the different species varied markedly in their responds to temperature. A. bifilosa showed a more definite trend of size variation with temperature than P. parvus and A. pacifica. Correlations of prosome length with salinity were significantly positive for almost all the small copepods. The relationship between chlorophyll concentration and prosome length was complicated for these copepods, but for P. parvus, chlorophyll concentration was also an important affecting factor. Furthermore, investigation needs to be done on food quality for some copepod. These results are essential to estimate the biomass and the production, and to understand these small copepods’ population dynamics in this human-affected bay.