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1.
A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco’s modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies. 相似文献
2.
Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell
culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells,
primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum.
The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25°C.
Supported by Doctoral Initial Fund of Ludong University (No.43304) 相似文献
3.
4.
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. 相似文献
5.
Effects of water temperature （17, 21, 25, 30 and 35℃） and body size （14.75-281.41 g initial body weight） on food consumption, growth, feed conversion, and dry matter content in orange-spotted grouper fed to satiation were investigated. The combined effect of temperature （T, ℃） and body weight （W, g） on maximum food consumption （Cmax, g/day） was described as： InCmax=-7.411＋0.828 InW＋0.317T4）.004 7T2, and the optimum feeding temperature was 33.9℃. The combined effect of temperature and body weight on growth （G） was described as： lnG=-4.461-0.2081nW＋0.394T-0.006 3T^2. The optimum growth temperature was 31.4℃, whereas overall growth rates were high at 25, 30 and 35 ℃. Feed conversion efficiencies （FCE, %）, increasing first and then decreasing with increasing temperature, averaged from 1.8 to 2.1 in terms of dry weight of food fish. The optimum temperature for FCE tended to be lower than that for growth or feeding. Dry matter content increased with both increasing water temperature （17, 25, 30 and 35℃） and body weight, and the combined effect of temperature and body weight on dry matter content （DM, %） was described as： lnDM =3.232＋0.01 4 lnW-0.004 4T＋0.001 2TInW. 相似文献
6.
We evaluated the effect of water temperature on the growth and physiology of the Chinese longsnout catfish (Leiocassis longirostris Günther). The fish were reared at four temperatures (20, 25, 30, and 35°C) and sampled on days 7, 20, and 30. We measured plasma levels of insulin, free thyroxine (FT4), free 3,5,3′-triiodothyronine (FT3), lysozyme and leukocyte phagocytic activity. The optimum water temperature for growth was 27.7°C. The plasma levels of insulin and FT4 declined significantly (P<0.05) on day 30 at temperatures above 20°C. Lysozyme activity was significantly (P<0.05) lower at 25°C than at other temperatures. We conclude that final weight, insulin, FT4, and lysozyme were significantly affected by water temperature. 相似文献
7.
Heterotrophic culture of microalgae to develop methods of increasing biomass productivity and storage lipids has brought new
insight to commercial biodiesel production. To understand the relationship between heterotrophy and lipid production, the
effects of carbon sources on the growth and lipid accumulation of Chlorella sorokiniana GXNN01 was studied. The alga exhibited an increased growth rate in response to the addition of carbon sources, which reached
the stationary phase after 48 h at 30°C. In addition, glucose and NaAc had a significant effect on the lipid accumulation
during the early-stationary phase. Specifically, the lipid content was 0.237±0.026 g g−1 cell dry weight and 0.272±0.041 g L−1 when glucose was used as the carbon source, whereas the lipid content reached 0.287±0.018 g g−1 cell dry weight and 0.288±0.008 g L−1 when NaAc was used as the carbon source. The neutral lipid content was found to first decrease and then increase over time
during the growth phase. A glucose concentration of 20 mmol L−1 gave the maximal lipid yield and the optimum harvest time was the early-stationary phase. 相似文献
8.
XU Yuyan GUO Huarong XIAO Qin SU Feng YIN Licheng 《中国海洋大学学报(英文版)》2007,6(4):369-372
The cytotoxicity of abamectin to the Gill Cell Line of Flounder (FG cell line) was examined in this study. It was found that the exposure of FG cells to abamectin caused the decreases of both cell growth rate and antioxidant enzyme activities,and the increase of intracellular O2- content. It was proposed that the reduction of antioxidant enzyme activities in FG cells caused the accu- mulation of O2- content in FG cells, leading to the change of cell morphology and even the death of cells. The results showed that FG cell line is suitable for the evaluation of the acute toxicity of abamectin. 相似文献
9.
LIU Yang LI Guiyang MO Zhaolan CHAI Zihan SHANG Anqi and MOU Haijin 《中国海洋大学学报(英文版)》2014,13(1):163-168
The bacteriophage P13 that infects Klebsiella serotype K13 contains a heat-stable depolymerase capable of effective degradation of exopolysaccharide(EPS) produced by this microorganism. In this study, the titer of phage P13, initially 2.0 × 107 pfu mL-1, was found increasing 20 min after infection and reached 5.0 × 109 pfu mL-1 in 60 min. Accordingly, the enzyme activity of depolymerase approached the maximum 60 min after infection. Treatment at 70℃ for 30 min inactivated all the phage, but retained over 90% of the depolymerase activity. Addition of acetone into the crude phage lysate led to precipitation of the protein, with a marked increase in bacterial EPS degradation activity and a rapid drop in the titer of phage. After partial purification by acetone precipitation and ultrafiltration centrifugation, the enzyme was separated from the phage particles, showing two components with enzyme activity on Q-Sepharose Fast Flow. The soluble enzyme had an optimum degradation activity at 60℃ and pH 6.5. Transmission electron microscopy demonstrated that the phage P13 particles were spherical with a diameter of 50 nm and a short stumpy tail. It was a double-strand DNA virus consisting of a nucleic acid molecule of 45976 bp. This work provides an efficient purification operation including thermal treatment and ultrafiltration centrifugation, to dissociate depolymerase from phage particles. The characterization of phage P13 and associated EPS depolymerase is beneficial for further application of this enzyme. 相似文献
10.
The FG-9307 cell line derived from the gill of flounder Paralichthys olivaceus was used in the present study to determine the acute cytotoxic effects of the organophosphorous pesticide, profenofos. It was found that the cell growth rate was markedly reduced by profenofos at the concentrations of 2.5, 5 and 7.5 mg/L tested; and that the fine structures of the cells were also altered by profenofos, as evidenced by dilation of nuclear membranes and mitochondria cristae, and presence of enlarged lysoscrneswith engulfed organelles and numerous vacuoles in the cytoplama. Probably, mitochondria, the cell ener-gy-generating sites, are the most prcrninent sites of profenofos cytotoxity in the cells. This seems to be the first report of the use of marine fish cell line for evaluation of the acute in vitro cytotoxicity of organophos-phorus pesticide. 相似文献
11.
Growth and energy budget of the polychaete, Neanthes japonica, at various temperatures (17, 20, 23, 26 and 29°) were investigated in this study. The growth, as indicated by final dry
weight and specific growth rate (SGR), increased with increasing temperature, with the maximum level at 26°C, and then decreased significantly at 29°C. A similar
trend was observed in feeding rate, food conversion efficiency (FCE) and apparent digestive rate (ADR). However, no significant differences were detected in ADR among all the temperature treatments. In the pattern of energy allocation, faeces energy was only a small component of energy
budget and had little influence on the proportion of food energy allocated to growth. The metabolic energy accounted for a
large portion of energy intake for each temperature treatment. The nitrogen excretion was appreciable with changing temperature.
The two expenditure terms (respiration energy and excretion energy) in energy budget were the major factors influencing the
proportion of food energy allocated to growth. These results revealed that temperature affected the growth of N. japonica mainly by influencing feeding rate and FCE. In addition, regression equations describing the relationship between feeding rate, faecal production, SGR, FCE and temperature were obtained. The optimum temperatures for feeding rate, FCE and SGR were estimated at 25.01°C, 24.24°C and 24.73°C, respectively, from the regression equations. 相似文献
12.
Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25°C. 相似文献
13.
Chong Peng Zhenghong Sui Wei Zhou Yiyi Hu Ping Mi Minjie Jiang Xiaodong Li Xudong Ruan 《中国海洋大学学报(英文版)》2018,17(3):623-631
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. 相似文献
14.
ＩＮＴＲＯＤＵＣＴＩＯＮＰｏｌｌｕｔｉｏｎｏｆｍａｒｉｎｅｅｃｏｓｙｓｔｅｍｐｏｓｅｓａｓｅｒｉｏｕｓｔｈｒｅａｔｔｏｍａｒｉｎｅｏｒｇａｎｉｓｍｓａｎｄｕｌｔｉｍａｔｅｌｙｔｈｅｅｎｔｉｒｅｅｃｏｓｙｓｔｅｍ.Ｉｎｒｅｃｅｎｔｙｅａｒｓ,ｏｒｇａｎｏｐｈｏｓｐｈｏｒｏｕｓ(ＯＰ)ｐｅｓｔｉｃｉｄｅｓｈａｖｅｌａｒｇｅｌｙｒｅｐｌａｃｅｄｏｒｇａｎｏｃｈｌｏｒｉｎｅｐｅｓｔｉｃｉｄｅｓｂｅｃａｕｓｅｏｆｔｈｅｉｒｒａｐｉｄｂｒｅａｋｄｏｗｎｉｎｗａｔｅｒａｎｄｔｈｅｉｒｌｏｗｅｎｖｉｒｏｎｍｅｎｔａ… 相似文献
15.
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. 相似文献
16.
Effects of basic fibroblast growth factor (bFGF) and insulin-like growth factor II (IGF-II) on cartilage cells from proboscis
of skate,Raja porasa Günther, were investigated in this study. The cartilage cells were cultured in 20% FBS-supplemented MEM medium at 24°C. Twelve hours
after culture initiation, the cartilage cells were treated with bFGF and IGF-II at different concentration combinations. It
was found that 20 ng/ml of bFGF or 80 ng/ml of IGF-II was enough to have obvious stimulating effect on the growth and division
of skate cartilage cells. Test of bFGF and IGF-II together, revealed that 20 ng/ml of bFGF and 80 ng/ml of IGF-II together
had the best stimulating effect on the growth and division of skate cartilage cells. The cartilage cells cultured could form
a monolayer at day 7.
This work was partially supported and funded by the China Education Committee as the “Imbursement Project for Studied Abroad
Returnees”. 相似文献
17.
Phytoplankton Assemblage of Yangtze River Estuary and the Adjacent East China Sea in Summer, 2004 总被引：6，自引：0，他引：6
A cruise was conducted from late August to early September 2004 with the intention of obtaining an interdisciplinary understanding of the Yangtze River Estuary including the biological, chemical and physical subjects. Water sample analysis indicated that total phytoplankton species richness was 137. Of them 81 were found in Bacillariophyta and 48 in Pyrrophyta, accounting for 59.1% and 35.0% respectively. The average cell abundance of surface water samples was 8.8×104 cells L-1, with the maximum, 102.9×104 cells L-1, encountered in the area (31.75°N, 122.33°E) and the minimum, 0.2×104 cells L-1, in (30.75°N, 122.17°E). The dominant species at most stations were Skeletonema costatum and Proboscia alata f. gracillima with the dominance of 0.35 and 0.27. Vertical distribution analysis indicated that obvious stratification of cell abundance and dominant species was found in the representative stations of 5, 18 and 33. Shannon-Wiener index and evenness of phytoplankton assemblage presented negative correlation with the cell abundance, with the optimum appearing in (30.75°N, 122.67°E). According to the PCA analysis of the environmental variables, elevated nutrients of nitrate, silicate and phosphate through river discharge were mainly responsible for the phytoplankton bloom in this area. 相似文献
18.
The effects of temperature and substrate on ontogenetic patterns and settlement behavior of bastard halibut, Paralichthys olivaceus （T. et S.） were studied in a series of laboratory experiments. Analysis on stage-based data revealed that except for settlement, and compared to the substrate, the ambient temperature dominantly controlled the hatch success and subsequent development of the species. The oosperm optimum survival rate of 60% during hatching occurred at 20℃, and survival rate of 29%, at 18℃ for larval rearing. The survival rate of larvae reared under variable temperature of 3.5-4.5℃ was higher than that of those reared under stable temperature. The maximum survival rate of 29%, corresponded to variation at temperature of 4℃. With increase in age newly settled juveniles preferred to lie on the fine substrate and gradually moved on the coarse substrate. GLM （general linear model） analysis showed that the combination of temperature and substrate had no significant impact on the survival of settling larvae, but definitely affected the duration of settlement. 相似文献
19.
In vitro acute cytotoxicity of neonicotinoid insecticide imidacloprid (IMI) to the gill cell line of flounder (FG) that collected in the gill of Paralichthys olivaceus, was examined by 3 widely used endpoint bioassays: NR (neutral red), MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) and TCP (total cell protein). The result shows that the IMI increased at concentrations ≥0.5 μg/ml. The IC50 value of NR, MTT, and TCP was 41.86, 38.46, and 39.08 μg/ml, respectively. The ultrastructural observation revealed that the mitochondria of the cells exposed to 60 μg/ml IMI for 48 h were severely damaged, swollen or disrupted, while their nuclei and rough endoplasmic reticulum (RER) remained normal. This would suggest that the mitochondria are probably the primary target of IMI. 相似文献
20.
Ｓｔｕｄｉｅｓｓｈｏｗｔｈａｔｔｈｅｇｌｏｂａｌｓｕｒｆａｃｅｔｅｍｐｅｒａｔｕｒｅｗｉｌｒｉｓｅａｔａｒａｔｅｏｆ０．３℃ｐｅｒ１０ａｉｎｔｈｅｎｅｘｔｓｅｖｅｒａｌｄｅｃａｄｅｓ（Ｈｏｕｇｈｔｏｎｅｔａｌ．，１９９０，１９９２；Ｓｈｉ，１９９５；... 相似文献