Catalase (CAT) and selenium-dependent glutathione peroxidase (Se-GPx) play a vital role in protecting organisms against various oxidative stresses by eliminating H202, The objective of this paper is to evaluate th...Catalase (CAT) and selenium-dependent glutathione peroxidase (Se-GPx) play a vital role in protecting organisms against various oxidative stresses by eliminating H202, The objective of this paper is to evaluate the roles of these antioxidant molecules in the ridgetail white prawn Exopalaemon carinicauda in response to low salinity stress. A complementary DNA (cDNA) containing the complete coding sequence of CAT was cloned from the hepatopancreas using reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The full-length cDNA of CAT (2 649 bp) contains a 5'-untranslated region (UTR) of 78 bp, a 3'- UTR of 1 017 bp, with a poly (A) tail, and an open reading frame of 1 554 bp encoding a 517-amino-acid polypeptide with predicted molecular mass of 58.46 kDa and estimated isoelectric point of 6.64. This CAT sequence contained the proximal active site signature (60FDRERIPERWHAKGAG76), proximal heme-ligand signature sequence (350RLFSYPDTH358) and three catalytic amino acid residues (His71, Asn144 and Tyr354). Sequence comparison showed that the CAT deduced amino acid sequence of E. carinicauda shared 68%-92% of identities with those of other species. Quantitative real-time PCR analysis revealed that CAT mRNA was widely expressed in the hepatopancreas (highest), hemocyte, eyestalk, heart, gill, muscle, ovary and stomach. Under low salinity stress, CAT and GPx mRNA expression levels both in the gill and hepatopancreas increased significantly at the first 48 h and 6 h respectively, indicating a tissue- and time-dependent antioxidant response in E. carinicauda. All these results indicate that E. carinicauda CAT is a member of the CAT family and might be involved in the acute response against low salinity stress.展开更多
The pattern recognition proteins(PRPs) play a major role in immune response of crustacean to resist pathogens.In the present study,as one of PRPs,lipopolysaccharide and β-1,3-glucan binding protein(LGBP) gene in the ...The pattern recognition proteins(PRPs) play a major role in immune response of crustacean to resist pathogens.In the present study,as one of PRPs,lipopolysaccharide and β-1,3-glucan binding protein(LGBP) gene in the ridge tail white prawn(Exopalaemon carinicauda)(Ec LGBP) was isolated.The full-length c DNA of Ec LGBP was 1338 bp,encoding a polypeptide of 366 amino acid residules.The deduced amino acid sequence of Ec LGBP shared high similarities with LGBP and BGBP from other crustaceans.Some conservative domains were predicted in Ec LGBP sequence.Ec LGBP constitutively expressed in most tissues at different levels,and the highest expression was observed in hepatopancreas.With infection time,the cumulative mortality increased gradually followed by the proliferation of Vibrio parahaemolyticus and white spot syndrome virus(WSSV).The expression of Ec LGBP in response to V.parahaemolyticus infection was up-regulated in hemocytes and hepatopancreas,and the up-regulation in hepatopancreas was earlier than that in hemocytes.Ec LGBP expression after WSSV infection increased at 3 h,then significantly decreased in both hemocytes and hepatopancreas.The results indicated that Ec LGBP was involved in the immune defense against bacterial and viral infections.展开更多
Exopalaemon carinicauda is one of the important economic shrimp species in China, and can tolerate a wide range of salinities. However, its disease resistance remains to be unclear in comparison with other shrimp spec...Exopalaemon carinicauda is one of the important economic shrimp species in China, and can tolerate a wide range of salinities. However, its disease resistance remains to be unclear in comparison with other shrimp species under salinity stress. In this study, the resistance to white spot syndrome virus(WSSV) of E. carinicauda and Litopenaeus vannamei was determined by comparing their hemocyanin(Hc) and phenoloxidase(PO) activities under different salinity stresses. In E. carinicauda, the PO activity and Hc gene transcript abundance showed a coherent pattern of increase and decrease while Hc content showed a slightly decrease with Vibrio anguillarum and WSSV infections. For both E. carinicauda and L. vannamei under salinity stress, the PO activity showed a positive correlation with the salinity while the Hc content and expression level of its gene increased significantly in salinities of 5, 15 and 25 g L^(-1). The survival rate of E. carinicauda with WSSV infection was higher than that of L. vannamei in the first 24 h under different salinity stresses. Drastic mortality of E.carinicauda and L. vannamei appeared at 48 h and 3 h post-injection, respectively. Furthermore, compared with L. vannamei, E. carinicauda displayed higher PO activity, Hc content and abundance of Hc gene m RNA. The results collectively indicated that Hc and PO have obviously functional connection in resisting pathogens and tolerating salinity stress, and PO activity and Hc gene m RNA abundance may reflect the resistance of shrimp to disease. E. carinicauda has higher level of immune potential than L. vannamei, suggesting its greater capacity in resisting pathogens under salinity stresses.展开更多
Fenneropenaeus chinensis, an economically important shrimp species, currently suffers from epizootic diseases due to high density stocking and bacterial infections. Oxytetracycline(OTC) has been widely used to treat v...Fenneropenaeus chinensis, an economically important shrimp species, currently suffers from epizootic diseases due to high density stocking and bacterial infections. Oxytetracycline(OTC) has been widely used to treat various systemic bacterial infections in shrimp farming. In the present study, the effect of gender on pharmacokinetics of OTC in F. chinensis was investigated. The OTC concentrations in hemolymph of shrimp after single intramuscular administration(75 mg OTC per kg body weight) were analyzed by high performance liquid chromatography and best described with a two-compartment open model which is characterized by a short elimination half-life, low clearance, and a relatively large apparent volume of distribution. The pharmacokinetic equations were Ct = 58.54e-0.38 t + 11.67e-0.04 t for females; and Ct = 27.94e-0.28 t + 14.87e-0.04 t for males. The distribution and elimination half-lives of OTC were 1.82 and 19.58 h, respectively, in females and 2.50 and 16.11 h, respectively, in males at 22℃. The areas under the drug concentration curve were 480 mg L-1 h-1 in females and 430 mg L-1 h-1 in males. The total body clearance of the drug was 157.11 m Lkg-1 h-1 in females and 172.47 m L kg-1 h-1 in males. The apparent volume distribution was 4.44 in females and 4.01 L kg-1 in males. There was no significant difference in pharmacokinetic parameters between female and male shrimps, indicating that there is no need to consider the gender effect in clinical use of OTC in F. chinensis farming.展开更多
This study examined the distribution and elimination of Norfloxacin(NFLX) in Fenneropenaeus chinensis ovary and egg and newly hatched larvae.Mature parental shrimp were exposed to 4 or 10 mg L 1NFLX for 2 or 5 d.Ovary...This study examined the distribution and elimination of Norfloxacin(NFLX) in Fenneropenaeus chinensis ovary and egg and newly hatched larvae.Mature parental shrimp were exposed to 4 or 10 mg L 1NFLX for 2 or 5 d.Ovary and eggs of the shrimp were sampled after spawning in order to detect NFLX residue using high-performance liquid chromatography(HPLC).Results showed that NFLX residue accumulated in F.chinensis eggs after the parental exposure,with the highest residue detected in ovary.To examine the fate of NFLX residue in larvae,we further determined the concentration of NFLX residue in F.chinensis eggs and larvae at 4 different developmental stages after 24-h exposure.From the newly metamorphosed larvae(0 h post-metamorphosis,h.p.m),samples were taken at different time intervals to 72 h.p.m.HPLC assay showed that the concentrations of NFLX residue in zoea exposed to 4 and 10 mg L 1NFLX were the highest at 1.5 h,i.e.,0.332 and 0.454 μg g 1,respectively.At the two NFLX exposure levels,the elimination time of half NFLX(half life) in nauplius was 45.36 and 49.85 h,respectively,followed by that in zoea(31.68 and 33.13 h),mysis larvae(42.24 and 47.28 h) and postlarvae(24.48 and 30.96 h).Both NFLX exposure levels had a germicidal effect.The distribution and elimination of NFLX residue in F.chinensis tissue,eggs and larvae correlated well with the drug exposure level.The disappearance of NFLX residue coincided with the larval growth,and the half-life of NFLX decreased with the larval development.展开更多
基金The Modern Agro-industry Technology Research System under contract No.CARS-47the National High-tech R&D Program(863 Program) of China under contract No.2012AA10A409+1 种基金the Special Fund for Independent Innovation of Shandong Province under contract No.2013CX80202the Special Fund for Agro-scientific Research in the Public Interest under contract No.201103034
文摘Catalase (CAT) and selenium-dependent glutathione peroxidase (Se-GPx) play a vital role in protecting organisms against various oxidative stresses by eliminating H202, The objective of this paper is to evaluate the roles of these antioxidant molecules in the ridgetail white prawn Exopalaemon carinicauda in response to low salinity stress. A complementary DNA (cDNA) containing the complete coding sequence of CAT was cloned from the hepatopancreas using reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The full-length cDNA of CAT (2 649 bp) contains a 5'-untranslated region (UTR) of 78 bp, a 3'- UTR of 1 017 bp, with a poly (A) tail, and an open reading frame of 1 554 bp encoding a 517-amino-acid polypeptide with predicted molecular mass of 58.46 kDa and estimated isoelectric point of 6.64. This CAT sequence contained the proximal active site signature (60FDRERIPERWHAKGAG76), proximal heme-ligand signature sequence (350RLFSYPDTH358) and three catalytic amino acid residues (His71, Asn144 and Tyr354). Sequence comparison showed that the CAT deduced amino acid sequence of E. carinicauda shared 68%-92% of identities with those of other species. Quantitative real-time PCR analysis revealed that CAT mRNA was widely expressed in the hepatopancreas (highest), hemocyte, eyestalk, heart, gill, muscle, ovary and stomach. Under low salinity stress, CAT and GPx mRNA expression levels both in the gill and hepatopancreas increased significantly at the first 48 h and 6 h respectively, indicating a tissue- and time-dependent antioxidant response in E. carinicauda. All these results indicate that E. carinicauda CAT is a member of the CAT family and might be involved in the acute response against low salinity stress.
基金supported by the earmarked fund for National ‘863’ Project of China (No.2012AA10A 409)Modern Agro-industry Technology Research System (No.CARS-47)+1 种基金Special Fund for Agro-scientific Research in the Public Interest (No.201103034)Independent Innovation Foundation of Shandong Province (No.2013CXC80202)
文摘The pattern recognition proteins(PRPs) play a major role in immune response of crustacean to resist pathogens.In the present study,as one of PRPs,lipopolysaccharide and β-1,3-glucan binding protein(LGBP) gene in the ridge tail white prawn(Exopalaemon carinicauda)(Ec LGBP) was isolated.The full-length c DNA of Ec LGBP was 1338 bp,encoding a polypeptide of 366 amino acid residules.The deduced amino acid sequence of Ec LGBP shared high similarities with LGBP and BGBP from other crustaceans.Some conservative domains were predicted in Ec LGBP sequence.Ec LGBP constitutively expressed in most tissues at different levels,and the highest expression was observed in hepatopancreas.With infection time,the cumulative mortality increased gradually followed by the proliferation of Vibrio parahaemolyticus and white spot syndrome virus(WSSV).The expression of Ec LGBP in response to V.parahaemolyticus infection was up-regulated in hemocytes and hepatopancreas,and the up-regulation in hepatopancreas was earlier than that in hemocytes.Ec LGBP expression after WSSV infection increased at 3 h,then significantly decreased in both hemocytes and hepatopancreas.The results indicated that Ec LGBP was involved in the immune defense against bacterial and viral infections.
基金supported by the earmarked fund for Modern Agro-Industry Technology Research System (No.CARS-47)the National Natural Science Foundation of China (No.31472275)+1 种基金the Program of Shandong Leading Talent (No.LNJY2015002)the Special Scientific Research Funds for Central Non-profit Institutes,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences (No.20603022016009)
文摘Exopalaemon carinicauda is one of the important economic shrimp species in China, and can tolerate a wide range of salinities. However, its disease resistance remains to be unclear in comparison with other shrimp species under salinity stress. In this study, the resistance to white spot syndrome virus(WSSV) of E. carinicauda and Litopenaeus vannamei was determined by comparing their hemocyanin(Hc) and phenoloxidase(PO) activities under different salinity stresses. In E. carinicauda, the PO activity and Hc gene transcript abundance showed a coherent pattern of increase and decrease while Hc content showed a slightly decrease with Vibrio anguillarum and WSSV infections. For both E. carinicauda and L. vannamei under salinity stress, the PO activity showed a positive correlation with the salinity while the Hc content and expression level of its gene increased significantly in salinities of 5, 15 and 25 g L^(-1). The survival rate of E. carinicauda with WSSV infection was higher than that of L. vannamei in the first 24 h under different salinity stresses. Drastic mortality of E.carinicauda and L. vannamei appeared at 48 h and 3 h post-injection, respectively. Furthermore, compared with L. vannamei, E. carinicauda displayed higher PO activity, Hc content and abundance of Hc gene m RNA. The results collectively indicated that Hc and PO have obviously functional connection in resisting pathogens and tolerating salinity stress, and PO activity and Hc gene m RNA abundance may reflect the resistance of shrimp to disease. E. carinicauda has higher level of immune potential than L. vannamei, suggesting its greater capacity in resisting pathogens under salinity stresses.
基金supported by the Earmarked Fund for Modern Agro-industry Technology Research System, China (No. CARS-47)the Special Fund for AgroScientific Research in the Public Interest of China (No. 201103034)
文摘Fenneropenaeus chinensis, an economically important shrimp species, currently suffers from epizootic diseases due to high density stocking and bacterial infections. Oxytetracycline(OTC) has been widely used to treat various systemic bacterial infections in shrimp farming. In the present study, the effect of gender on pharmacokinetics of OTC in F. chinensis was investigated. The OTC concentrations in hemolymph of shrimp after single intramuscular administration(75 mg OTC per kg body weight) were analyzed by high performance liquid chromatography and best described with a two-compartment open model which is characterized by a short elimination half-life, low clearance, and a relatively large apparent volume of distribution. The pharmacokinetic equations were Ct = 58.54e-0.38 t + 11.67e-0.04 t for females; and Ct = 27.94e-0.28 t + 14.87e-0.04 t for males. The distribution and elimination half-lives of OTC were 1.82 and 19.58 h, respectively, in females and 2.50 and 16.11 h, respectively, in males at 22℃. The areas under the drug concentration curve were 480 mg L-1 h-1 in females and 430 mg L-1 h-1 in males. The total body clearance of the drug was 157.11 m Lkg-1 h-1 in females and 172.47 m L kg-1 h-1 in males. The apparent volume distribution was 4.44 in females and 4.01 L kg-1 in males. There was no significant difference in pharmacokinetic parameters between female and male shrimps, indicating that there is no need to consider the gender effect in clinical use of OTC in F. chinensis farming.
基金supported by the Earmarked Fund for Modern Agro-industry Technology Research System,China(No.CARS-47)the Special Fund for Agro-scientific Research in the Public Interest of China(No.201103034)
文摘This study examined the distribution and elimination of Norfloxacin(NFLX) in Fenneropenaeus chinensis ovary and egg and newly hatched larvae.Mature parental shrimp were exposed to 4 or 10 mg L 1NFLX for 2 or 5 d.Ovary and eggs of the shrimp were sampled after spawning in order to detect NFLX residue using high-performance liquid chromatography(HPLC).Results showed that NFLX residue accumulated in F.chinensis eggs after the parental exposure,with the highest residue detected in ovary.To examine the fate of NFLX residue in larvae,we further determined the concentration of NFLX residue in F.chinensis eggs and larvae at 4 different developmental stages after 24-h exposure.From the newly metamorphosed larvae(0 h post-metamorphosis,h.p.m),samples were taken at different time intervals to 72 h.p.m.HPLC assay showed that the concentrations of NFLX residue in zoea exposed to 4 and 10 mg L 1NFLX were the highest at 1.5 h,i.e.,0.332 and 0.454 μg g 1,respectively.At the two NFLX exposure levels,the elimination time of half NFLX(half life) in nauplius was 45.36 and 49.85 h,respectively,followed by that in zoea(31.68 and 33.13 h),mysis larvae(42.24 and 47.28 h) and postlarvae(24.48 and 30.96 h).Both NFLX exposure levels had a germicidal effect.The distribution and elimination of NFLX residue in F.chinensis tissue,eggs and larvae correlated well with the drug exposure level.The disappearance of NFLX residue coincided with the larval growth,and the half-life of NFLX decreased with the larval development.