Low temperature stress on the hematological parameters and <Emphasis Type="Italic">HSP</Emphasis> gene expression in the turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis>

To study the effect of low temperature stress on hematological parameters and HSP gene expression in the turbot (Scophthalmus maximus), water temperature was lowered rapidly from 18 to 1°C. During the cooling process, three individuals were removed from culture tanks at 18, 13, 8, 5, 3, and 1°C. Blood samples and tissues were taken from each individual, hematological indices and HSP gene expression in tissues were measured. The red blood cell count, white blood cell count, and hemoglobin concentration decreased significantly (P < 0.05) as temperature decreased. Enzyme activities of plasma alanine transaminase and creatine kinase increased as temperature decreased, whereas aspartic transaminase and γ-glutamyl transpeptidase activities displayed no obvious changes above 1°C and lactate dehydrogenase activity increased first and then decreased. Blood urea nitrogen and uric acid levels were highest at 8°C, and creatinine concentration was highest at 3°C. The concentrations of plasma cortisol, cholesterol, and triglyceride all increased significantly (P < 0.05) as temperature decreased. The serum glucose concentration increased first and then decreased to the initial level. The HSP70 mRNA expression showed various patterns in different tissues, whereas HSP90 mRNA expression showed the same tendency in all tissues. Overall, these results indicate that temperature decreases in the range of 8 to 5°C may induce a stress response in S. maximus and that temperature should be kept above 8°C in the aquaculture setting to avoid damage to the fish.     

Compensatory growth of juvenile brown flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> following low temperature treatment for different periods

We investigated the effects of low temperature (8.5°C) on the growth and feeding rates and feed conversion efficiency of juvenile P. olivaceus with an average initial weight of 3.87 ± 0.06 g (mean ± SE). Fish were exposed to 8.5°C for 0 (control), 1, 2, 3 and 4 weeks, and then to 20°C for 10, 9, 8, 7 and 6 weeks, respectively. Low temperature clearly led to growth depression. The weight of fish exposed to low temperature for 1 week was restored to that of control, while that of fish exposed to low temperature longer was significantly decreased (P < 0.05). During the entire low-temperature period, specific growth rate, feeding rate and feed conversion efficiency of the fish were significantly lower (P < 0.05) than those of control, while in the whole recovery period, specific growth and average feeding rate were markedly higher (P < 0.05) than those of control. At the end of experiment, only the feeding rate of the fish exposed to low temperature for 1 week was not significantly different from that of control (P > 0.05). Feeding rate and feed conversion efficiency were reduced at low temperature in juvenile P. olivaceus. The compensatory growth of juvenile P. olivaceus may therefore be attributed to the improvement of feeding rate. Our results suggested that growth depression occurs when juvenile P. olivaceus are exposed to low temperature for more than one week.     

Functional characterization of a novel microbial esterase identified from the Indian Ocean and its use in the stereoselective preparation of (<Emphasis Type="Italic">R</Emphasis>)-methyl mandelate

Genomic mining has identified a novel microbial alkaline esterase from the Indian Ocean. This esterase was overexpressed in E. coli BL21 (DE3) and further functionally characterized. Under optimal conditions (10 mmol/L substrate, pH 6.0, 2 h at 40 °C), this esterase can hydrolyze racemic methyl mandelate to (R)-methyl mandelate with very high optical purity (e.e. >99%) and yield (nearly 90%). Interestingly, the stereoselectivity of this esterase is opposite to that of two previously reported lipases that can generate (S)-methyl mandelate through the hydrolysis of racemic methyl mandelate. No organic solvents or other additives were required to optimize the optical purity and production of the final chiral product (R)-methyl mandelate, which can potentially simplify the production procedure of (R)-methyl mandelate catalyzed by esterase.     

Analysis of DNA Methylation of <Emphasis Type="Italic">Gracilariopsis lemaneiformis</Emphasis> Under Temperature Stress Using the Methylation Sensitive Amplification Polymorphism (MSAP) Technique

Gracilariopsis lemaneiformis is an economically important agarophyte, which contains high quality gel and shows a high growth rate. Wild population of G. lemaneiformis displayed resident divergence, though with a low genetic diversity as was revealed by amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses. In addition, different strains of G. lemaneiformis are diverse in morphology. The highly inconsistence between genetic background and physiological characteristics recommends strongly to the regulation at epigenetic level. In this study, the DNA methylation change in G. lemaneiformis among different generation branches and under different temperature stresses was assessed using methylation sensitive amplified polymorphism (MSAP) technique. It was shown that DNA methylation level among different generation branches was diverse. The full and total methylated DNA level was the lowest in the second generation branch and the highest in the third generation. The total methylation level was 61.11%, 60.88% and 64.12% at 15°C, 22°C and 26°C, respectively. Compared with the control group (22°C), the fully methylated and totally methylated ratios were increased in both experiment groups (15°C and 26°C). All of the cytosine methylation/demethylation transform (CMDT) was further analyzed. High temperature treatment could induce more CMDT than low temperature treatment did.     

Interspecific competition and allelopathic interaction between <Emphasis Type="Italic">Karenia mikimotoi</Emphasis> and <Emphasis Type="Italic">Dunaliella salina</Emphasis> in laboratory culture

Algal allelopathy is a manifold ecological/physiological phenomenon that is focused on chemical interactions and autotoxicity. We investigated the allelopathic interactions between Karenia mikimotoi and Dunaliella salina in laboratory cultures based on diff erent temperature (15°C, 20°C, and 25°C) and lighting (40, 80, and 160 μmol/(m²·s)) conditions. The growth of D. salina in bi-algae culture (1:1 size/density) was significantly restrained. The results of cell-free filtrate culture indicate that direct cell-tocell contact was not necessary in interspecific competition. Further experimental results demonstrated that allelochemicals released from K. mikimotoi were markedly influenced by both temperature (P =0.013) and irradiance (P =0.003), resulting in diff erent growth characteristics of D. salina in filtrate mediums. Compared with the plateau period, K. mikimotoi exudates in the exponential phase had a stronger short-term inhibition effect on D. salina in normal conditions. A clear concentration-dependent relationship was observed in the effect of allelochemicals released from K. mikimotoi with low-promoting and high-repressing effects on D. Salina in a short time-scale. In addition, allelopathic substances remain stable and effective under high temperature and pressure stress. Many flocculent sediments adhering with D. salina cells were observed in all filtrate mediums, while the quantity and color depended on the original culture conditions.     

Diel variation in metabolism and ammonia excretion of <Emphasis Type="Italic">Marphysa sanguinea</Emphasis> (Polychaeta: Eunicidae)

Polychaetes provide an excellent food resource for fish and represent the dominant zoobenthos in marine ecosystems. Diel variation in the rates of metabolism and ammonia-N excretion of Marphysa sanguinea were studied. The worms were grouped according to their wet body weight into small (S; 1.24±0.06 g), medium (M; 4.00±0.30 g), and large (L; 8.54±1.08 g) categories. Their weight-specific metabolic rates, based on aerobic respiration (R), were measured at 16°C (±0.2°C) and classed as either routine (R _R) or standard (R _S) rates. Both respiration types decreased with increasing body weight. Respiration was described by R = a W ^b, where b was -0.400 9 and -0.532 0 for R _R and R _S, respectively. Diurnal changes in R _S for each group was relatively flat, with a slightly increasing trend with time, but was relatively stable as a whole. R _R of the diurnal variation of worms was higher than R S, but both had similar overall trends. The peak values of specific dynamic action (SDA) (R _SDA) in the S, M, and L groups were 2.704, 1.149, and 0.682 mg/(g?h), respectively. The durations of SDA were 13, 6, and 6 h, respectively and the energy expenditures of SDA were 377.98, 117.34, and 74.94 J/g, respectively. These data indicate that the metabolic rates were higher in smaller individuals, which is advantageous for their rapid growth.     

Feeding broodstocks different starfish diets affect growth and survival of larvae of trumpet shell (<Emphasis Type="Italic">Charonia lampas sauliae</Emphasis> Reeve 1844)

Trumpet shell (Charonia lampas sauliae) (Mollusca, Heterogastropoda, Cymatidae) has extensive economic value. Studies on the artificial larval development of C. lampas sauliae for aquaculture utilization have become especially important due to the finite natural resources. In the present study, the growth and survival rate of the larvae of C. lampas sauliae broodstocks fed three types of starfish diets, Asterina pectinifera Müller & Troschel 1842, A. amurensis Lütken 1871 and their mixture were compared. The larval size increased gradually between day 10 and day 20 after hatching at 15°C and 20°C. No difference was found in body size and specific growth rate (SGR) (two-way ANOVA; P > 0.05). However, during transition from trochophore to veliger stage 20 days after hatching, significant increases in larval survival and growth rates were observed. The maximum survival rate was observed on day 10. The mean survival rate was 0.463, 0.730 and 0.515 at 15°C, and 0.369, 0.713 and 0.444 at 20°C when A. pectinifera, A. amurensis and their mixture were fed, respectively. The SGR and survival rate of the larvae were definitely influenced by the diets (P < 0.05), and the effect of A. amurensis alone was higher than that of A. pectinifera alone and their mixture.     

Inbreeding depression on growth and survival of full-sib family of Manila clam (<Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>)

In present study, the inbreeding depression (ID) of growth and survival of Manila clam (Ruditapes philippinarum) was investigated at larval and juvenile stages. Nine inbred families (A ₂, B ₂, C ₂, D ₂, E ₂, F ₂, G ₂, H ₂ and I ₂) were established by mating within nine full-sib families with expected inbreeding coefficient of 0.25. Inbred families showed significant differences in shell length and hatching rate of D-larvae (straight-hinged larvae). The larvae of the nine inbred families grew slower than those of control group (CG), and their ID value ranged from 0.81% ± 6.09% to 16.10% ± 1.49%. The ID value of larval survival rate varied between 27.47% ± 9.36% and 70.50% ± 13.66%. The ID was also detected for juvenile growth in A ₂, B ₂, C ₂, and D ₂, which ranged from 4.60 ± 2.21 to 17.71 ± 7.73. The A ₂ family maintained the highest juvenile survival rate, whereas the other inbred families exhibited ID values varying between 62.79% ± 4.54% and 96.14% ± 0.87%. The linear relationship of estimated ID between growth and survival was negatively correlated (R = ?0.434, P < 0.05). The results of this study suggested that the ID of growth was common at the larval stage but was less prevalent at juvenile stage. In contrast, the ID of survival increased from larval to juvenile stage. A better understanding of the effect of inbreeding may aid to selective breeding of Manila clam.     

Optimization of protectant,salinity and freezing condition for freeze-drying preservation of <Emphasis Type="Italic">Edwardsiella tarda</Emphasis>

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 NaCl 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°C or ?40°C 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.     

Distribution of dominant zooplankton species along a latitudinal gradient in China sea during spring

Dominant species of zooplankton community vary with latitude. Though China possesses a vast coastal area in northwestern Pacific, studies on the latitudinal dominant species gradient are rare. We collected zooplankton samples from Haizhou Bay(34.56?–35.19?N, 119.51?–120.30?E), Yueqing Bay(28.14?–28.38?N, 121.10?–121.21?E) and Dongshan Bay(23.65?–23.90?N, 117.45?–117.60?E) in May 2012 and May 2013 to preliminarily characterize the latitudinal dominant species distribution. All the samples were collected vertically using a 0.505 mm mesh plankton net with 0.8 m in mouth diameter from bottom to surface. Calanus sinicus, Aidanosagitta crassa, Labidocera euchaeta, Zonosagitta nagae, Acartia pacifica and Paracalanus parvus were found to be dominant. C. sinicus was the most dominant species and the unique one occurred in all three bays. With latitude decreasing, both the abundance and proportion of C. sinicus declined sharply. Cluster analysis showed that the 6 dominant species could be divided into 3 groups, based on their occurrences in the three bays. Our results suggested that the distribution of dominant species along the coast of China has a significant latitudinal gradient. C. sinicus which widely distributes in the coastal water of the northwestern Pacific can well adapt to the temperature at different latitudes. The high abundance in Haizhou Bay indicated that C. sinicus was an exemplary warm-temperate species, and more commonly occurs in the north of China seas. The ecological characteristics of dominant species change from warm-temperate type in high-latitudinal bays to warm water type in low-latitudinal bays.     

Global water vapor content decreases from 2003 to 2012: An analysis based on MODIS data

Water vapor in the earth′s upper atmosphere plays a crucial role in the radiative balance, hydrological process, and climate change. Based on the latest moderate-resolution imaging spectroradiometer(MODIS) data, this study probes the spatio-temporal variations of global water vapor content in the past decade. It is found that overall the global water vapor content declined from 2003 to 2012(slope b = –0.0149, R = 0.893, P = 0.0005). The decreasing trend over the ocean surface(b = –0.0170, R = 0.908, P = 0.0003) is more explicit than that over terrestrial surface(b = –0.0100, R = 0.782, P = 0.0070), more significant over the Northern Hemisphere(b = –0.0175, R = 0.923, P = 0.0001) than that over the Southern Hemisphere(b = –0.0123, R = 0.826, P = 0.0030). In addition, the analytical results indicate that water vapor content are decreasing obviously between latitude of 36°N and 36°S(b = 0.0224, R = 0.892, P = 0.0005), especially between latitude of 0°N and 36°N(b = 0.0263, R = 0.931, P = 0.0001), while the water vapor concentrations are increasing slightly in the Arctic regions(b = 0.0028, R = 0.612, P = 0.0590). The decreasing and spatial variation of water vapor content regulates the effects of carbon dioxide which is the main reason of the trend in global surface temperatures becoming nearly flat since the late 1990 s. The spatio-temporal variations of water vapor content also affect the growth and spatial distribution of global vegetation which also regulates the global surface temperature change, and the climate change is mainly caused by the earth's orbit position in the solar and galaxy system. A big data model based on gravitational-magmatic change with the solar or the galactic system is proposed to be built for analyzing how the earth's orbit position in the solar and galaxy system affects spatio-temporal variations of global water vapor content, vegetation and temperature at large spatio-temporal scale. This comprehensive examination of water vapor changes promises a holistic understanding of the global climate change and potential underlying mechanisms.     

Trophic Interaction in a <Emphasis Type="Italic">Portunus rituberculatus</Emphasis> Polyculture Ecosystem Based on Carbon and Nitrogen Stable Isotope Analysis

Trophic interaction among various biomass groups in a swimming crab Portunus trituberculatus polyculture pond was investigated using carbon and nitrogen stable isotope analysis. The polycultured animal species also included white shrimp Litopenaeus vannamei, short-necked clam Ruditapes philippinarum, and redlip mullet Liza haematochila. The mean δ¹³C value for all the biomass groups in polyculture ecosystem ranged from ?25.61‰ to ?16.60‰, and the mean δ¹⁵N value ranged from 6.80‰ to 13.09‰. Significant difference in the δ¹³C value was found between particulate organic matter (POM) and sediment organic matter (SOM) (P < 0.05), indicating that these two organic matter pools have different material sources. Assuming that a ¹³C-enrichment factor of 1.00‰ and a ¹⁵N-enrichment factor of 2.70‰ existed between consumer and prey, diets of the four cultured animals were estimated using a stable isotope mixing model. The estimated model results indicated that P. trituberculatus mainly feed on Aloidis laevis; L. vannamei mainly feed on shrimp feed; while A. laevis, R. philippinarum and L. haematochelia mainly feed on POM. Shrimp feed was also an important food source of R. philippinarum and L. haematochelia. The diets of P. trituberculatus, L. vannamei, R. philippinarum, and L. haematochila showed complementary effects in this polyculture ecosystem. Our finding indicated that the polyculture of these four organisms with suitable farming density could make an effective use of most of the food sources, which can make a highly efficient polyculture ecosystem.     

Effects of Anthropogenic Disturbance on Sediment Organic Carbon Mineralization Under Different Water Conditions in Coastal Wetland of a Subtropical Estuary

The changes in soil organic carbon(C) mineralization as affected by anthropogenic disturbance directly determine the role of soils as C source or sink in the global C budget. The objectives of this study were to investigate the effects of anthropogenic disturbance(aquaculture pond, pollutant discharge and agricultural activity) on soil organic C mineralization under different water conditions in the Minjiang River estuary wetland, Southeast China. The results showed that the organic C mineralization in the wetland soils was significantly affected by human disturbance and water conditions(P 0.001), and the interaction between human disturbance activities and water conditions was also significant(P 0.01). The C mineralization rate and the cumulative mineralized carbon dioxide-carbon(CO_2-C)(at the 49th day) ranked from highest to lowest as follows: Phragmites australis wetland soil aquaculture pond sediment soil near the discharge outlet rice paddy soil. This indicated that human disturbance inhibited the mineralization of C in soils of the Minjiang River estuary wetland, and the inhibition increased with the intensity of human disturbance. The data for cumulative mineralized CO_2-C showed a good fit(R~2 0.91) to the first-order kinetic model C_t = C_0(1 – exp(–kt)). The kinetic parameters C_0, k and C_0 k were significantly affected by human disturbance and water conditions. In addition, the total amount of mineralized C(in 49 d) was positively related to C_0, C_0 k and electrical conductivity of soils. These findings indicated that anthropogenic disturbance suppressed the organic C mineralization potential in subtropical coastal wetland soils, and changes of water pattern as affected by human activities in the future would have a strong influence on C cycling in the subtropical estuarine wetlands.     

The community structure and seasonal dynamics of plankton in Bange Lake,northern Tibet,China

The seasonal variations in biomass, abundance, and species composition of plankton in relation to hydrography were studied in the saline Bange Lake, northern Tibet, China. Sampling was carried out between one to three times per month from May 2001 to July 2002. Salinity ranged from 14 to 146. The air and water temperature exhibited a clear seasonal pattern, and mean annual temperatures were approximately 4.8°C and 7.3°C, respectively. The lowest water temperature occurred in winter from December to March at-2°C and the highest in June and July at 17.7°C. Forty-one phytoplankton taxa, 21 zooplankton, and 5 benthic or facultative zooplankton were identifi ed. The predominant phytoplankton species were Gloeothece linearis, Oscillatoria tenuis, Gloeocapsa punctata, Ctenocladus circinnatus, Dunaliella salina, and Spirulina major. The predominant zooplankton species included H olophrya actra, Brachionus plicatilis, Daphniopsis tibetana, Cletocamptus dertersi, and A rctodiaptomus salinus. The mean annual total phytoplankton density and biomass for the entire lake were 4.52×10~7 cells/L and 1.60 mg/L, respectively. The annual mean zooplankton abundance was 52, 162, 322, and 57, 144 ind./L, in the three sublakes. The annual mean total zooplankton biomass in Lakes 1–3 was 1.23, 9.98, and 2.13 mg/L, respectively. The annual mean tychoplankton abundances in Bg1, 2, and 3 were 47, 67, and 654 ind./L. The annual mean tychoplankton biomass was 2.36, 0.16, and 2.03 mg/L, respectively. The zooplankton biomass(including tychoplankton) in the lake was 9.11 mg/L. The total number of plankton species in the salt lake was signifi cantly negatively correlated with salinity.     

Seasonal variation and principle of cyanobacterial biomass and forms in the water source area of Chaohu City,China

We investigated seasonal variations in cyanobacterial biomass and the forms of its dominant population(M. aeruginosa) and their correlation with environmental factors in the water source area of Chaohu City,China from December 2011 to October 2012. The results show that species belonging to the phylum Cyanophyta occupied the maximum proportion of phytoplankton biomass,and that the dominant population in the water source area of Chaohu City was M. aeruginosa. The variation in cyanobacterial biomass from March to August 2012 was well fitted to the logistic growth model. The growth rate of cyanobacteria was the highest in June,and the biomass of cyanobacteria reached a maximum in August. From February to March 2012,the main form of M.aeruginosa was the single-cell form; M.aeruginosa colonies began to appear from April,and blooms appeared on the water surface in May. The maximum diameter of the colonies was recorded in July,and then gradually decreased from August. The diameter range of M. aeruginosa colonies was 18.37–237.77 μm,and most of the colonies were distributed in the range 20–200 μm,comprising 95.5% of the total number of samples. Temperature and photosynthetically active radiation may be the most important factors that influenced the annual variation in M. aeruginosa biomass and forms. The suitable temperature for cyanobacterial growth was in the range of 15–30°C. In natural water bodies,photosynthetically active radiation had a significant positive influence on the colonial diameter of M.aeruginosa(P0.01).     

Photosynthetic response of <Emphasis Type="Italic">Scirpus validus</Emphasis> and <Emphasis Type="Italic">Typha orientalis</Emphasis> to elevated temperatures in Dianchi Lake,Southwestern China

Understanding the effects of simulated warming on photosynthetic performance of aquatic plants may provide strong supports for predicting future dynamics of wetland ecosystems in the context of climate change. The plateau wetlands located in Yunnan province are highly sensitive to climate warming due to their high altitude and cold temperature. Here, we conducted a temperaturecontrolled experiment using two temperature manipulations (ambient temperature as the control and 2°C higher than ambient temperature as the warmed treatment) to determine the photosynthetic characteristics of two lakeside dominant species (Scirpus validus Vahl and Typha orientalis C. Presl.) in Dianchi Lake. Net photosynthetic rate (P_n), stomatal conductance (G_s), intercellular CO₂ concentration (C_i), and transpiration rate of S. validus that grew under warmed treatment were all significantly higher than those under the control. G_s and C_i of T. orientalis showed similar patterns as S. validus did. For the response curves of P_n to photosynthetic active radiation (Pn-PAR) and intercellular CO₂ concentration (P_n-C_i), S. validus had higher P_n values under elevated temperatures than the control, while P_n-PAR and P_n-C_i curves of T. orientalis did not separate clearly under two temperature scenarios. Both S. validus and T. orientalis had higher maximum net photosynthetic rate, light saturation point, dark respiration rate, the maximum rate of RuBP carboxylation (V_cmax), maximum electron transport rate driving RuBP regeneration (J_max), the ratio of V_cmax to J_max, triosephosphate utilization, and 1, 5-bishosphate carboxylase ribulose content under warmed treatment than those under the control. This study provides a preliminary step for predicting the future primary production and vegetation dynamics of plateau wetlands in Yunnan province.     

Biodegradation of <Emphasis Type="Italic">Enteromorpha</Emphasis> polysaccharides by intestinal micro-community from <Emphasis Type="Italic">Siganus oramin</Emphasis>

Micro-communities are supposed to have more potential functions of biodegradation of polysaccharides than single strain; however, the intestinal micro-communities involved in the biodegradation of Enteromorpha polysaccharides (EP) were seldom reported. In order to obtain the EP-degrading micro-community, the intestines of Siganus oramin was obtained to isolate the micro-communities, which were enriched by 0.3% of EP as the sole carbon source. A stable micro-community with EP degradative capability was achieved after seven generations of subculture, named H1. Results showed that H1 was able to degrade 75% of EP within 24 hours, and the activity of EP lyases reached 500 U mL^?1 in 32 hours. With denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analysis, ten bacteria closely related to Marinomonas pontica, Microbacterium sp., Leucobacter chironomi, Cyclobacterium sp., Algoriphagus winogradskyi, Pseudoalteromonas sp. and Vibrio sp. were determined. Furthermore, compared with the DGGE bands sequence and the clone library analysis, the dominant bacteria of the EP-biodegrading micro- community were Pseudoalteromonas sp. and Vibrio sp., with the respective proportion of 38% and 46%, and they should play an important role in EP degradation together with other degrading bacteria in the micro-community H1.     

Temporal and spatial variations of abundance of phycocyanin- and phycoerythrin-rich <Emphasis Type="Italic">Synechococcus</Emphasis> in Pearl River Estuary and adjacent coastal area

Three surveys were carried out in Pearl River Estuary and adjacent coastal area in May, August, and November, 2013, to investigate the temporal and spatial variations of abundance of phycoerythrin-rich Synechococcus (PE-rich SYN) and phycocyanin-rich Synechococcus (PC-rich SYN). The effects of environmental factors on the alternation of the different Synechococcus groups were also elucidated. PE-rich SYN was detected in three surveys, whereas PC-rich SYN was detected in May and August, but not in November. The highest abundances of PE-rich SYN and PC-rich SYN were recorded in August and May, with mean values of 74.17×10³ and 189.92×10³ cells mL^?1, respectively. From May to November, the relative abundance of PE-rich SYN increased, whereas that of PC-rich SYN declined. PE-rich and PC-rich SYN presented similar horizontal distributions with high abundance in the southern estuary in May, and in the western estuary in August. The abundances of PE-rich and PC-rich SYN were high at 27–32°C and salinity of 10–20. PC-rich SYN was not detected at < 24°C, and PC:PE-rich SYN decreased in abundance with salinity increase. When less than 20 mg L^?1, suspended particulate matter (SPM) was helpful for Synechococcus growth. PE-rich SYN decreased in abundance when the concentration of dissolved inorganic nitrogen increased in May and November, and the concentration of phosphate increased in November. However, PC-rich SYN abundance and nutrients showed no correlation. Principal component analysis and regression analysis indicated that PE-rich SYN significantly correlated with the principal components that were affected by environmental factors.     

Establishment and characterization of a testicular Sertoli cell line from olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

The culture of Sertoli cells has become an indispensable resource in studying spermatogenesis. A new Sertoli cell line (POSC) that consisted predominantly of fibroblast-like cells was derived from the testis of the olive flounder Paralichthys olivaceus and sub-cultured for 48 passages. Analysis of the mtDNA COI gene partial sequence confirmed that the cell line was from P. olivaceus. Cells were optimally maintained at 25°C in DMEM/F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, and epidermal growth factor. The growth curve of POSC showed a typical “S” shape. Chromosome analysis revealed that the cell line possessed the normal P. olivaceus diploid karyotype of 2n=48t. POSC expressed dmrt1 but not vasa, which was detected using RT-PCR and sequencing. Immunocytochemistry revealed that the cells exhibited the testicular Sertoli cell marker FasL. Therefore, POSC appeared to consist of testicular Sertoli cells. Bright fluorescent signals were observed after the cells were transfected with pEGFP-N3 plasmid, with the transfection efficiency reaching 10%. This research not only offers an ideal model for further gene expression and regulation studies on P. olivaceus, but also serves as valuable material in studying fish spermatogenesis, Sertoli cell-germ cell interactions, and the mechanism of growth and development of testis.