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 fr...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.展开更多
Olaquindox(OLA), one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial ent...Olaquindox(OLA), one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial enteritis. In this study, we evaluated the effect of dietary OLA on the growth of large yellow croaker(Pseudosciaena crocea R.) and the histological distribution of OLA and its metabolite 3-methyl-quinoxaline-2-carboxylic acid(MQCA) in fish tissues. Four diets containing 0(control), 42.5, 89.5 and 277.2 mg kg-1 OLA, respectively, were formulated and tested, 3 cages(1.0 m × 1.0 m × 1.5 m) each diet and 100 juveniles(9.75 ± 0.35 g) each cage. The fish were fed to satiation twice a day at 05:00 am and 17:00 pm for 8 weeks. The survival rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet containing 0 and 277.2 mg kg-1 OLA(P < 0.05), while the weight gain rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet without OLA(control)(P<0.05), but similar to that of fish fed the diet with 277.2 mg kg-1 OLA. Fish fed the diet with 277.2 mg kg-1 OLA had the highest content of OLA and MQCA in liver(3.44 and 0.39 mg kg-1, respectively), skin(0.46 and 0.09 mg kg-1, respectively) and muscle(0.24 and 0.06 mg kg-1, respectively). In average, fish fed the diet containing OLA had the highest content of OLA and MQCA in liver which was followed by skin and muscle(P < 0.05), whereas OLA and MQCA were not detectable in control. Our findings demonstrated that OLA and MQCA accumulated in large yellow croaker when it was fed with the diet containing OLA, thus imposing a potential safety risk to human health.展开更多
L ethal temperature tolerance was determined for about 8 cm, age 0 Pseudosciaena crocea using both slow heating and rapid transfer protocol. The acclimatization temperature was 28 ℃ with summer season, lethal tempera...L ethal temperature tolerance was determined for about 8 cm, age 0 Pseudosciaena crocea using both slow heating and rapid transfer protocol. The acclimatization temperature was 28 ℃ with summer season, lethal temperature ( LT50 value ) of slow heating protocol ( CTMax ) was 35.0 ℃, and the upper and lower incipient lethal temperatures of rapid transfer protocol were 34.2 ℃ and 17.5 ℃ respectively.展开更多
The present study investigated conditions for inducing mito-gynogenetic(endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea.In haploid control groups,the development of eggs ...The present study investigated conditions for inducing mito-gynogenetic(endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea.In haploid control groups,the development of eggs was activated with ultraviolet radiated semen.All fry presented typical haploid syndrome in the haploid control groups,and were verified as haploids using cytometry.After hydrostatic pressure treatment,morphologically normal fry reappeared at different frequencies according to the intensity and time of pressure shock.Fry with normal appearance in the pressure treated groups were verified as gynogenetic double haploids(GDHs),containing only one allele from the female parent at all four diagnostic microsatellite loci.For a fixed duration of 3 min,the optimal intensity of blocking the first mitosis was determined to be 40 Mpa,which was similar to that of blocking the second meiosis.There was a "window" of starting time,from 36.1 min to 38.1 min post-insemination at 25.0±1.0°C,within which the production of GDHs was not significantly different.Maximum production of morphologically normal fries,9.36%±2.97% of developed eggs,was found when the eggs were shocked with hydrostatic pressure at 40 Mpa for 3 min,starting from 38.1 min post insemination at 25.0±1.0°C.展开更多
A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus (LYCIV) is described, which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated fr...A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus (LYCIV) is described, which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen. Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus (RSIV) and sea bass iridovirus (SBIV), suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone. The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes, the expected fragment was detected from spleen DNA samples of infected fishes, whereas no fragments were amplified from healthy fish spleen DNA, white spot syndrome baculoviruses (WSBV) DNA and pseudorabies virus (PRV) DNA. Detection limit of this method was 10(-7) ng positive plasmid DNA containing target sequence, equal to about 100 virions. In the infected experiment, first positive detection (1/4) appeared at Day 3 post-infection, all fish (4/4) tested positive at Day 7, however obvious symptoms were observed at Day 8, so LYCIV infection could be detected prior to the appearance of obvious symptoms. These results indicate that this PCR method could be used for early, rapid and specific detection of LYCIV infection.展开更多
This study performs the quantitative analysis and comparison to acoustic signal characteristics of Large yellow croaker （Pseudosciaena crocea） at two different ages. The sounds were recorded from the fishes in a net...This study performs the quantitative analysis and comparison to acoustic signal characteristics of Large yellow croaker （Pseudosciaena crocea） at two different ages. The sounds were recorded from the fishes in a net-cage. Two exponential oscillation functions are built to fit the acoustic signal of the fishes. The signal characteristic of the oscillation frequency and attenuation coefficient was described quantitatively. Simulation curves of the function could fit well acoustic signals. Both the average oscillation frequency and attenuation coefficient of the fitted signals from the 13-15-month-old fishes are lower than those from the 7-8-month-old fishes. The results suggest that the oscillation frdquency and attenuation coefficient of the acoustic signal flmction may be relevant to the physical process of sound production and age characteristics of Large yellow croaker. This study may be valuable for the acoustic application to the artificial culture of the species.展开更多
基金supported by grants from the National High Technology Research and Development Program ('863' Program) of China (Grant Nos. 2006AA10A401 and 2006AA09Z406)
文摘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.
基金supported by the National Key Technologies R & D Program for the 10th and 11th Five-year Plan of China (2001BA505B-06 2006BAD03B03)the Program for Changjiang Scholars and Innovative Research Team in University
文摘Olaquindox(OLA), one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial enteritis. In this study, we evaluated the effect of dietary OLA on the growth of large yellow croaker(Pseudosciaena crocea R.) and the histological distribution of OLA and its metabolite 3-methyl-quinoxaline-2-carboxylic acid(MQCA) in fish tissues. Four diets containing 0(control), 42.5, 89.5 and 277.2 mg kg-1 OLA, respectively, were formulated and tested, 3 cages(1.0 m × 1.0 m × 1.5 m) each diet and 100 juveniles(9.75 ± 0.35 g) each cage. The fish were fed to satiation twice a day at 05:00 am and 17:00 pm for 8 weeks. The survival rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet containing 0 and 277.2 mg kg-1 OLA(P < 0.05), while the weight gain rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet without OLA(control)(P<0.05), but similar to that of fish fed the diet with 277.2 mg kg-1 OLA. Fish fed the diet with 277.2 mg kg-1 OLA had the highest content of OLA and MQCA in liver(3.44 and 0.39 mg kg-1, respectively), skin(0.46 and 0.09 mg kg-1, respectively) and muscle(0.24 and 0.06 mg kg-1, respectively). In average, fish fed the diet containing OLA had the highest content of OLA and MQCA in liver which was followed by skin and muscle(P < 0.05), whereas OLA and MQCA were not detectable in control. Our findings demonstrated that OLA and MQCA accumulated in large yellow croaker when it was fed with the diet containing OLA, thus imposing a potential safety risk to human health.
基金supported by the Science and Technology Bureau of Zhejiang Province(No.2003C33064)the Science and Technology Bureau of Ningbo City(No.2003C10002)the Key Laboratory of Marine and Estuarine Fisheries of the Ministry of Agriculture.
文摘L ethal temperature tolerance was determined for about 8 cm, age 0 Pseudosciaena crocea using both slow heating and rapid transfer protocol. The acclimatization temperature was 28 ℃ with summer season, lethal temperature ( LT50 value ) of slow heating protocol ( CTMax ) was 35.0 ℃, and the upper and lower incipient lethal temperatures of rapid transfer protocol were 34.2 ℃ and 17.5 ℃ respectively.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2006AA10A405)the Foundation for Innovative Research Team of Jimei University(No.2006A001),the Science Foundation of Jimei University(No.ZQ2006037)
文摘The present study investigated conditions for inducing mito-gynogenetic(endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea.In haploid control groups,the development of eggs was activated with ultraviolet radiated semen.All fry presented typical haploid syndrome in the haploid control groups,and were verified as haploids using cytometry.After hydrostatic pressure treatment,morphologically normal fry reappeared at different frequencies according to the intensity and time of pressure shock.Fry with normal appearance in the pressure treated groups were verified as gynogenetic double haploids(GDHs),containing only one allele from the female parent at all four diagnostic microsatellite loci.For a fixed duration of 3 min,the optimal intensity of blocking the first mitosis was determined to be 40 Mpa,which was similar to that of blocking the second meiosis.There was a "window" of starting time,from 36.1 min to 38.1 min post-insemination at 25.0±1.0°C,within which the production of GDHs was not significantly different.Maximum production of morphologically normal fries,9.36%±2.97% of developed eggs,was found when the eggs were shocked with hydrostatic pressure at 40 Mpa for 3 min,starting from 38.1 min post insemination at 25.0±1.0°C.
文摘A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus (LYCIV) is described, which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen. Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus (RSIV) and sea bass iridovirus (SBIV), suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone. The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes, the expected fragment was detected from spleen DNA samples of infected fishes, whereas no fragments were amplified from healthy fish spleen DNA, white spot syndrome baculoviruses (WSBV) DNA and pseudorabies virus (PRV) DNA. Detection limit of this method was 10(-7) ng positive plasmid DNA containing target sequence, equal to about 100 virions. In the infected experiment, first positive detection (1/4) appeared at Day 3 post-infection, all fish (4/4) tested positive at Day 7, however obvious symptoms were observed at Day 8, so LYCIV infection could be detected prior to the appearance of obvious symptoms. These results indicate that this PCR method could be used for early, rapid and specific detection of LYCIV infection.
基金supported by the National Natural Science Foundation of China(41276040,11174240)the Fujian Province Natural Science Fund Project(2060203)the Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry
文摘This study performs the quantitative analysis and comparison to acoustic signal characteristics of Large yellow croaker （Pseudosciaena crocea） at two different ages. The sounds were recorded from the fishes in a net-cage. Two exponential oscillation functions are built to fit the acoustic signal of the fishes. The signal characteristic of the oscillation frequency and attenuation coefficient was described quantitatively. Simulation curves of the function could fit well acoustic signals. Both the average oscillation frequency and attenuation coefficient of the fitted signals from the 13-15-month-old fishes are lower than those from the 7-8-month-old fishes. The results suggest that the oscillation frdquency and attenuation coefficient of the acoustic signal flmction may be relevant to the physical process of sound production and age characteristics of Large yellow croaker. This study may be valuable for the acoustic application to the artificial culture of the species.
