Flounder gill （FG） cells were used to isolate lymphocystis disease virus （LCDV） and two monoclonal antibodies （Mabs） （1A8 and 3G3） against LCDV were used to trace LCDV infection to FG cells. FG monolayer cells...Flounder gill （FG） cells were used to isolate lymphocystis disease virus （LCDV） and two monoclonal antibodies （Mabs） （1A8 and 3G3） against LCDV were used to trace LCDV infection to FG cells. FG monolayer cells was inoculated with LCDV supernatant, obtained from lymphocystis cells of diseased flounder, Paralichthys olivaceus. LCDV infection was detected with Mabs employing immunocytochemical assay （ICA） and indirect immunofluorescence assay test （IIFAT） technique. Detected by IIFAT, they were specifie for LCDV. The results of experimental infection illustrated that FG cells was sensitive to LCDV, and showed virus-infection positive detected by ICA. Cytopathic effect （CPE） occurred 1-2 days post inoculation （PI）, and half tissue culture infection dosage （TCID50） of vires supematant was 2^2.57 per 40μl. Tracing by IIFAT showed that LCDV positive signal first appeared at the cell membrane immediately PI, and then in cytoplasm at 24h PI, it reached the strongest positive at 48-72 h PI, and began to decrease at 96h PI.展开更多
In this study, we determined the cytotoxicity and genotoxicity of carbamate insecticide carbaryl to flounder gill （FG） cells and its teratogenicity to zebrafish embryos. The cytotoxicity of carbaryl to FG cells was ...In this study, we determined the cytotoxicity and genotoxicity of carbamate insecticide carbaryl to flounder gill （FG） cells and its teratogenicity to zebrafish embryos. The cytotoxicity of carbaryl to FG cells was determined with methods including MTT and neutral red uptaking （NRU）, lactate dehydrogenase （LDH） releasing and Hoechst 33342 and propidiurn idodide （PI） double staining. Moderate cytotoxicity in a concentration-dependent manner was observed. The 24h-IC50 value of 53.48 ± 1.21, 59.13 ± 1.19 and 46.21 ± 1.24 mgL 1 carbaryl was obtained through MTT, NRU and LDH assays, respectively. Double fluorescence staining dem- onstrated that carbaryl induced the death of FG cells mainly through necrosis. There was no significant genotoxicity found in the FG cells exposed to the highest testing concentration of carbaryl （20mgL-1, P〉0.05） as was demonstrated by Comet assay. Zebrafish embryos exposed to carbaryl at concentrations 〉10 mg L-1 displayed moderate toxic effects on the survival, spontaneous movement, hatching, heart rates of the embryos and their development, which were evidenced by yolk and pericardial sac edemas, body length reduction and tail flexure in time- and concentration-dependent manners at specific stages. The 24h-, 48h- and 96h-LC50 values of carbaryl to zebra fish embryos were 41.80 ± 1.10, 17.80 ± 1.04 and 14.46 ± 1.05 mg L-1, respectively. These results suggested that car- baryl is moderately toxic to FG ceils cultured in vitro and zebrafish embryos, and the FG cells were similar to zebrafish embryos in their sensitivity to carbaryl as 24h-IC50 and LC50 indicated.展开更多
Rita rita exposed to a concentration of 6.9 mg per liter(96-h LC_(50)of an anionic detergent, dodecylbenzene sodium sulfonate)exhibited a gradual decrease in the lipid moieties of the epi- thelial cells,club cells,and...Rita rita exposed to a concentration of 6.9 mg per liter(96-h LC_(50)of an anionic detergent, dodecylbenzene sodium sulfonate)exhibited a gradual decrease in the lipid moieties of the epi- thelial cells,club cells,and goblet mucous cells lining the gill arch and gill filament epithelium. However,in time,no reaction of any of the lipid moieties could be observed,indicating the absence of the same,using various histochemical techniques.The results are discussed in light of the mechanistic understanding of detergent action.1989 Academic Press,Inc.展开更多
探讨大菱鲆病毒疫苗的研制,以牙鲆鳃细胞(FG细胞)为增殖体系,利用细胞病变效应(CPE)为判断指标,对大菱鲆红体病虹彩病毒(Turbot reddish body iridovirus,TRBIV)的体外最适增殖条件等进行了研究。结果表明,病毒接种量、病毒吸附时间、...探讨大菱鲆病毒疫苗的研制,以牙鲆鳃细胞(FG细胞)为增殖体系,利用细胞病变效应(CPE)为判断指标,对大菱鲆红体病虹彩病毒(Turbot reddish body iridovirus,TRBIV)的体外最适增殖条件等进行了研究。结果表明,病毒接种量、病毒吸附时间、细胞培养条件对TRBIV的体外增殖均有明显的影响,以0.2 mL病毒液接种FG细胞、吸附2 h、用含10%胎牛血清的MEM培养液于22℃培养时,病毒的增殖速度最快。研究还发现,病毒的收获时间和病毒液冻融次数对所得病毒的感染力均有显著影响,在最佳增殖条件下,病毒在FG细胞中增殖至第4天时进行收获,且只冻融1次,其感染活性最强,反复冻融后病毒的感染活性会显著降低。展开更多
文摘Flounder gill （FG） cells were used to isolate lymphocystis disease virus （LCDV） and two monoclonal antibodies （Mabs） （1A8 and 3G3） against LCDV were used to trace LCDV infection to FG cells. FG monolayer cells was inoculated with LCDV supernatant, obtained from lymphocystis cells of diseased flounder, Paralichthys olivaceus. LCDV infection was detected with Mabs employing immunocytochemical assay （ICA） and indirect immunofluorescence assay test （IIFAT） technique. Detected by IIFAT, they were specifie for LCDV. The results of experimental infection illustrated that FG cells was sensitive to LCDV, and showed virus-infection positive detected by ICA. Cytopathic effect （CPE） occurred 1-2 days post inoculation （PI）, and half tissue culture infection dosage （TCID50） of vires supematant was 2^2.57 per 40μl. Tracing by IIFAT showed that LCDV positive signal first appeared at the cell membrane immediately PI, and then in cytoplasm at 24h PI, it reached the strongest positive at 48-72 h PI, and began to decrease at 96h PI.
基金supported by the grants from National High-tech R&D Program of China (863 Program, Grant No. 2012AA 10A402)National Natural Science Foundation of China (Grant Nos. 31172391 and 31472274)+1 种基金Key Laboratory Open Foundation of Marine Bioactive Substances and Modern Analytical Technology, State Oceanic Administration (No. MBSMAT-2011-01)Fundamental Research Funds for the Central Universities of China (Grant No. 201122005)
文摘In this study, we determined the cytotoxicity and genotoxicity of carbamate insecticide carbaryl to flounder gill （FG） cells and its teratogenicity to zebrafish embryos. The cytotoxicity of carbaryl to FG cells was determined with methods including MTT and neutral red uptaking （NRU）, lactate dehydrogenase （LDH） releasing and Hoechst 33342 and propidiurn idodide （PI） double staining. Moderate cytotoxicity in a concentration-dependent manner was observed. The 24h-IC50 value of 53.48 ± 1.21, 59.13 ± 1.19 and 46.21 ± 1.24 mgL 1 carbaryl was obtained through MTT, NRU and LDH assays, respectively. Double fluorescence staining dem- onstrated that carbaryl induced the death of FG cells mainly through necrosis. There was no significant genotoxicity found in the FG cells exposed to the highest testing concentration of carbaryl （20mgL-1, P〉0.05） as was demonstrated by Comet assay. Zebrafish embryos exposed to carbaryl at concentrations 〉10 mg L-1 displayed moderate toxic effects on the survival, spontaneous movement, hatching, heart rates of the embryos and their development, which were evidenced by yolk and pericardial sac edemas, body length reduction and tail flexure in time- and concentration-dependent manners at specific stages. The 24h-, 48h- and 96h-LC50 values of carbaryl to zebra fish embryos were 41.80 ± 1.10, 17.80 ± 1.04 and 14.46 ± 1.05 mg L-1, respectively. These results suggested that car- baryl is moderately toxic to FG ceils cultured in vitro and zebrafish embryos, and the FG cells were similar to zebrafish embryos in their sensitivity to carbaryl as 24h-IC50 and LC50 indicated.
文摘Rita rita exposed to a concentration of 6.9 mg per liter(96-h LC_(50)of an anionic detergent, dodecylbenzene sodium sulfonate)exhibited a gradual decrease in the lipid moieties of the epi- thelial cells,club cells,and goblet mucous cells lining the gill arch and gill filament epithelium. However,in time,no reaction of any of the lipid moieties could be observed,indicating the absence of the same,using various histochemical techniques.The results are discussed in light of the mechanistic understanding of detergent action.1989 Academic Press,Inc.
