In this paper, ontogeny of immune-related organs during theearly development of carp was studied by histochemical technique-lcineBlue staining under acid conditions and PAS reaction(AB-PAS staining). Thekidney appeare...In this paper, ontogeny of immune-related organs during theearly development of carp was studied by histochemical technique-lcineBlue staining under acid conditions and PAS reaction(AB-PAS staining). Thekidney appeared one day before hatching, spleen, liver and pancreas emergedon the same day of hatching, and thymus was not found until the third day afterhatching. Ontogeny of these immune related organs of the carp in thisresearch is earlier than that reported by Botham.展开更多
[Objective] The paper was to investigate the protective effects of different concentrations of Bacillus subtilis peptidoglycan(PG) on β-conglycinin-induced inflammatory injury in intestinal epithelial cells of juveni...[Objective] The paper was to investigate the protective effects of different concentrations of Bacillus subtilis peptidoglycan(PG) on β-conglycinin-induced inflammatory injury in intestinal epithelial cells of juvenile carp(Cyprinus carpio).[Method] In 24-cell microplates, the intestinal epithelial cells(IECs) of juvenile carp were primarily cultured for 72 h at 26°C and 6% CO2, and then the IECs were randomly divided into6 groups with 4 replicates per group. One of the six groups was set as negative control group, and the other groups were all supplemented with 1.0 mg/mL β-conglycinin in culture medium to establish inflammatory injury. At 24 h post induction, the culture media were changed into B. subtilis PG culture media with the concentrations of 0(positive control group), 0.15, 0.30, 0.45 and 0.60 mg/mL, respectively. The samples were collected to measure the antioxidant and anti-inflammatory indices at 12, 24 and 36 h post culture.[Result]β-conglycinin exposure significantly decreased the activity of ASA, AHR, SOD, CAT, GPx, and increased the PC content and the mRNA expression of inflammatory cytokines(IL-1β, IL-8, TNF-α1,IL-10 and TGF-β). At 12, 24 and 36 h post PG treatment, the activities of ASA, AHR, SOD, CAT, GPx and the content of PC in cells decreased in a dose-dependent manner;the mRNA levels of IL-1β, IL-8 and TNF-α1 were down-regulated and those of IL-10 and TGF-β were up-regulated.[Conclusion] Different concentrations of B. subtilis PG could protect IECs oxidative damage induced by β-conglycinin and improve the antioxidant capacity of IECs. High concentration of PG could improve the anti-inflammatory ability of IECs by inhibiting inflammatory factors and promoting the gene expression of anti-inflammatory cytokines.展开更多
Isolated ovarian tissues from the common carp, Cyprinus carpio were incubated in vitro to obtain a discrete effect of four common toxicants of industrial origin, namely phenol, sulfide, mercuric chloride and cadmium c...Isolated ovarian tissues from the common carp, Cyprinus carpio were incubated in vitro to obtain a discrete effect of four common toxicants of industrial origin, namely phenol, sulfide, mercuric chloride and cadmium chloride, on gonadotropin-induced alteration of nonesterified and esterified cholesterol and steroidogenic enzymes, △5-3β-HSD and 17β-HSD activity. Stage II ovarian tissue containing 30-40% mature oocytes were shown to be most responsive to gonadotropins in depleting only nonesterified cholesterol moiety and stimulating the activity of both. Safe doses of above mentioned toxicants when added separately to stage II ovarian tissue with oLH (1 μg/incubation) gonadotropin-induced depletion of nonesterified cholesterol and gonadotropin-induced stimulation of the activity of both enzymes was significantly inhibited. Esterified cholesterol remained almost unaltered. Findings clearly indicate the impairment of gonadotropin induced fish ovarian steroidogenesis by the four toxicants separately.展开更多
Gonadal function in fish, CJprinus carpio was significantly affected by sublethal doses of mercuric chloride (HgCl2 ) and cadmium chloride (CdCl2 ) in chronic (45 days ) exposure.Parameters investigated were nonesteri...Gonadal function in fish, CJprinus carpio was significantly affected by sublethal doses of mercuric chloride (HgCl2 ) and cadmium chloride (CdCl2 ) in chronic (45 days ) exposure.Parameters investigated were nonesterified (NE) and esterified (E) cholestcrol of ovary,liver and serum and ovarian 3β-Hydroxysteroid and 17β-Hydroxysteroid dehyrogenaseenzyme activity and serum and pituitary gonadotropin (GtH) levels. Both the pollutantswere able to reduce the hypothalamic extract (HE) or gonadotropin releasing hormone(GnRH) induced pituitary GtH release in vitro. Short term (96h) exposure of the fish tothe pollutants had no significant effect on the gonadal function. ln addition to thedeleterious effect of pollutants on the gonadal steroidogenesis and pituitary gonadotropin release, using [4-14C] cholesterol as a tracer it was found that for 45 days expeure, HgCl2'had an adverse effect on the transport of cholesterol from circulation to ovary.展开更多
【目的】青田田鱼在浅水环境中经常面临急性低温胁迫,探究青田田鱼在急性低温和复温环境下的生理响应机制,为青田田鱼科学越冬和耐低温品种选育提供理论依据。【方法】通过液相色谱—质谱(LC-MS)非靶向代谢组学技术,提取9.0℃胁迫6 h(CO...【目的】青田田鱼在浅水环境中经常面临急性低温胁迫,探究青田田鱼在急性低温和复温环境下的生理响应机制,为青田田鱼科学越冬和耐低温品种选育提供理论依据。【方法】通过液相色谱—质谱(LC-MS)非靶向代谢组学技术,提取9.0℃胁迫6 h(CO组)、升温至28.0℃复温恢复6 h(RE组)和28.0℃对照组(Con组)青田田鱼鳃组织样本代谢峰。预处理后的代谢组数据与数据库匹配鉴定代谢物种类。通过主成分分析(PCA)和正交最小偏二乘判别分析(OPLS-DA)鉴定代谢组学数据的可靠性。根据Student’s test检验的P值(P<0.05)和OPLS-DA模型得到的变量权重值(VIP>1)筛选组间差异代谢物。对差异代谢物进行KEGG信号通路富集分析,并用Fisher精确检验分析筛选与急性低温和复温最相关的代谢通路。对重要代谢通路制作聚类热图,显示组间差异和代谢物水平变化趋势。【结果】PCA和OPLS-DA分析结果显示,组内样本聚集,组间样本分离,组间的代谢物水平存在显著差异。OPLS-DA得分图显示各组Q2累计值均在0.5以上,R2Y累积值均接近1.0,模型具有较高的解释度和可靠性;共鉴定到1222个代谢物,其中CO vs Con,RE vs CO和RE vs Con分别筛选鉴定出232、238和300种显著差异代谢物(P<0.05)。各组间差异代谢物主要富集在花生四烯酸代谢、甘油磷脂代谢、类固醇激素生物合成和嘌呤代谢等代谢途径;急性低温胁迫下,皮质醇水平上调,而花生四烯酸及其代谢物和甘油磷脂代谢物下调。复温后花生四烯酸、甘油磷脂代谢物和嘌呤代谢物相对Con组下调,皮质醇含量下调,牛磺酸含量上调。【结论】急性低温可导致青田田鱼免疫、物质转运和信号传递等功能异常。皮质醇的合成和代谢可能是青田田鱼抵抗急性低温胁迫的重要机制。复温后青田田鱼鳃组织损伤在短期内无法得到完全恢复。牛磺酸合成可能是青田田鱼在复温过程中的重要恢复机制。展开更多
Gonadotropin-releasing hormone (GnRH) is a conservative neurodecapeptide fam-ily, which plays a crucial role in regulating the gonad development and in controlling the final sexual maturation in vertebrate. Two differ...Gonadotropin-releasing hormone (GnRH) is a conservative neurodecapeptide fam-ily, which plays a crucial role in regulating the gonad development and in controlling the final sexual maturation in vertebrate. Two differing cGnRH-II cDNAs of common carp, namely cGnRH-II cDNA1 and cDNA2, were firstly cloned from the brain by rapid amplification of cDNA end (RACE) and reverse transcription- polymerase chain reaction (RT-PCR). The length of cGnRH-II cDNA1 and cDNA2 was 622 and 578 base pairs (bp), respectively. The cGnRH-II pre-cursors encoded by two cDNAs consisted of 86 amino acids, including a signal peptide, cGnRH-II decapeptide and a GnRH-associated peptide (GAP) linked by a Gly-Lys-Arg proteolytic site. The results of intron trapping and Southern blot showed that two differing cGnRH-II genes in common carp genome were further identified, and that two genes might exist as a single copy. The multi-gene coding of common carp cGnRH-II gene offered novel evidence for gene duplica-tion hypothesis. Using semi-quantitative RT-PCR, expression and relative expression levels of cGnRH-II genes were detected in five dissected brain regions, pituitary and gonad of common carp. With the exception of no mRNA2 in ovary, two cGnRH-II genes could be expressed in all the detected tissues. However, expression levels showed an apparent difference in different brain regions, pituitary and gonad. According to the expression characterization of cGnRH-II genes in brain areas, it was presumed that cGnRH-II might mainly work as the neurotransmitter and neuromodulator and also operate in the regulation for the GnRH releasing. Then, the ex-pression of cGnRH-II genes in pituitary and gonad suggested that cGnRH-II might act as the autocrine or paracrine regulator.展开更多
To test the effectiveness of introducing live zooplankton against direct manuring in ornamental fish ponds upon their survival and production, larvae of koi carp, Cyprinus carpio L., were cultured for 11 weeks in eart...To test the effectiveness of introducing live zooplankton against direct manuring in ornamental fish ponds upon their survival and production, larvae of koi carp, Cyprinus carpio L., were cultured for 11 weeks in earthen ponds maintained according to four management regimes： （1） live zooplankton fed to carp larvae （LF）; （2） direct fertilization with poultry manure （PM）; （3） direct fertilization with cowdung （CD）; and （4） a control treatment （C）. There were three replicates for each treatment. The growth of heterotrophic bacteria and pathogenic microorganisms like Aeromonas sp. and Pseudomonas sp. were also examined in response to pond management. Values of dissolved oxygen were significantly higher （P〈0.05） in the water of LF ponds, compared to other treatments, while the PM and CD treatments recorded were significantly higher （P〈0.05） values of PO4 - P, NH4 - N, NO3 - N, NO2 - N, specific conductivity, alkalinity, and BOD, compared to the LF and C treatments. The percentages of organic carbon and total nitrogen in the bottom sediments were higher in the PM and CD treatments compared to LF （P〈0.05）. Average counts of heterotrophic bacteria in the water of PM and CD ponds were significantly higher than other treatments （P〈0.05）. The development of Aeromonas sp. and Pseudomonas sp. were significantly higher （P〈0.05） in the PM and CD treatments. Weight gain of koi carp stocked in LF was significantly higher （P〈0.05） than that of fish in the other treatments. There was a significant difference in the survival rate of koi carp among the treatments ranging from 67.21% in C to 90.11% in LF. The results suggest that raising koi carp larvae in ponds and feeding them exogenously with zooplankton would support high rates of survival and production through maintenance of better water quality and greater abundance of zooplankton in the system. Significantly lower abundance of Aeromonas sp. and Pseudomonas sp. in the LF treatment considerably lowered any possibility of occurrence of bacterial disease.展开更多
文摘In this paper, ontogeny of immune-related organs during theearly development of carp was studied by histochemical technique-lcineBlue staining under acid conditions and PAS reaction(AB-PAS staining). Thekidney appeared one day before hatching, spleen, liver and pancreas emergedon the same day of hatching, and thymus was not found until the third day afterhatching. Ontogeny of these immune related organs of the carp in thisresearch is earlier than that reported by Botham.
基金Supported by Science and Technology Research Project of Henan Province(172102110205)
文摘[Objective] The paper was to investigate the protective effects of different concentrations of Bacillus subtilis peptidoglycan(PG) on β-conglycinin-induced inflammatory injury in intestinal epithelial cells of juvenile carp(Cyprinus carpio).[Method] In 24-cell microplates, the intestinal epithelial cells(IECs) of juvenile carp were primarily cultured for 72 h at 26°C and 6% CO2, and then the IECs were randomly divided into6 groups with 4 replicates per group. One of the six groups was set as negative control group, and the other groups were all supplemented with 1.0 mg/mL β-conglycinin in culture medium to establish inflammatory injury. At 24 h post induction, the culture media were changed into B. subtilis PG culture media with the concentrations of 0(positive control group), 0.15, 0.30, 0.45 and 0.60 mg/mL, respectively. The samples were collected to measure the antioxidant and anti-inflammatory indices at 12, 24 and 36 h post culture.[Result]β-conglycinin exposure significantly decreased the activity of ASA, AHR, SOD, CAT, GPx, and increased the PC content and the mRNA expression of inflammatory cytokines(IL-1β, IL-8, TNF-α1,IL-10 and TGF-β). At 12, 24 and 36 h post PG treatment, the activities of ASA, AHR, SOD, CAT, GPx and the content of PC in cells decreased in a dose-dependent manner;the mRNA levels of IL-1β, IL-8 and TNF-α1 were down-regulated and those of IL-10 and TGF-β were up-regulated.[Conclusion] Different concentrations of B. subtilis PG could protect IECs oxidative damage induced by β-conglycinin and improve the antioxidant capacity of IECs. High concentration of PG could improve the anti-inflammatory ability of IECs by inhibiting inflammatory factors and promoting the gene expression of anti-inflammatory cytokines.
文摘Isolated ovarian tissues from the common carp, Cyprinus carpio were incubated in vitro to obtain a discrete effect of four common toxicants of industrial origin, namely phenol, sulfide, mercuric chloride and cadmium chloride, on gonadotropin-induced alteration of nonesterified and esterified cholesterol and steroidogenic enzymes, △5-3β-HSD and 17β-HSD activity. Stage II ovarian tissue containing 30-40% mature oocytes were shown to be most responsive to gonadotropins in depleting only nonesterified cholesterol moiety and stimulating the activity of both. Safe doses of above mentioned toxicants when added separately to stage II ovarian tissue with oLH (1 μg/incubation) gonadotropin-induced depletion of nonesterified cholesterol and gonadotropin-induced stimulation of the activity of both enzymes was significantly inhibited. Esterified cholesterol remained almost unaltered. Findings clearly indicate the impairment of gonadotropin induced fish ovarian steroidogenesis by the four toxicants separately.
文摘Gonadal function in fish, CJprinus carpio was significantly affected by sublethal doses of mercuric chloride (HgCl2 ) and cadmium chloride (CdCl2 ) in chronic (45 days ) exposure.Parameters investigated were nonesterified (NE) and esterified (E) cholestcrol of ovary,liver and serum and ovarian 3β-Hydroxysteroid and 17β-Hydroxysteroid dehyrogenaseenzyme activity and serum and pituitary gonadotropin (GtH) levels. Both the pollutantswere able to reduce the hypothalamic extract (HE) or gonadotropin releasing hormone(GnRH) induced pituitary GtH release in vitro. Short term (96h) exposure of the fish tothe pollutants had no significant effect on the gonadal function. ln addition to thedeleterious effect of pollutants on the gonadal steroidogenesis and pituitary gonadotropin release, using [4-14C] cholesterol as a tracer it was found that for 45 days expeure, HgCl2'had an adverse effect on the transport of cholesterol from circulation to ovary.
文摘【目的】青田田鱼在浅水环境中经常面临急性低温胁迫,探究青田田鱼在急性低温和复温环境下的生理响应机制,为青田田鱼科学越冬和耐低温品种选育提供理论依据。【方法】通过液相色谱—质谱(LC-MS)非靶向代谢组学技术,提取9.0℃胁迫6 h(CO组)、升温至28.0℃复温恢复6 h(RE组)和28.0℃对照组(Con组)青田田鱼鳃组织样本代谢峰。预处理后的代谢组数据与数据库匹配鉴定代谢物种类。通过主成分分析(PCA)和正交最小偏二乘判别分析(OPLS-DA)鉴定代谢组学数据的可靠性。根据Student’s test检验的P值(P<0.05)和OPLS-DA模型得到的变量权重值(VIP>1)筛选组间差异代谢物。对差异代谢物进行KEGG信号通路富集分析,并用Fisher精确检验分析筛选与急性低温和复温最相关的代谢通路。对重要代谢通路制作聚类热图,显示组间差异和代谢物水平变化趋势。【结果】PCA和OPLS-DA分析结果显示,组内样本聚集,组间样本分离,组间的代谢物水平存在显著差异。OPLS-DA得分图显示各组Q2累计值均在0.5以上,R2Y累积值均接近1.0,模型具有较高的解释度和可靠性;共鉴定到1222个代谢物,其中CO vs Con,RE vs CO和RE vs Con分别筛选鉴定出232、238和300种显著差异代谢物(P<0.05)。各组间差异代谢物主要富集在花生四烯酸代谢、甘油磷脂代谢、类固醇激素生物合成和嘌呤代谢等代谢途径;急性低温胁迫下,皮质醇水平上调,而花生四烯酸及其代谢物和甘油磷脂代谢物下调。复温后花生四烯酸、甘油磷脂代谢物和嘌呤代谢物相对Con组下调,皮质醇含量下调,牛磺酸含量上调。【结论】急性低温可导致青田田鱼免疫、物质转运和信号传递等功能异常。皮质醇的合成和代谢可能是青田田鱼抵抗急性低温胁迫的重要机制。复温后青田田鱼鳃组织损伤在短期内无法得到完全恢复。牛磺酸合成可能是青田田鱼在复温过程中的重要恢复机制。
基金This work was supported by the'863'High Technology Project of China(Grant No.2001AA213101,2001AA212281)the National Natural Science Foundation of China(Grant No.200102006)+1 种基金the'973'Project of the Ministry of ScienceTechnology(Grant No.2001 CB 109006).
文摘Gonadotropin-releasing hormone (GnRH) is a conservative neurodecapeptide fam-ily, which plays a crucial role in regulating the gonad development and in controlling the final sexual maturation in vertebrate. Two differing cGnRH-II cDNAs of common carp, namely cGnRH-II cDNA1 and cDNA2, were firstly cloned from the brain by rapid amplification of cDNA end (RACE) and reverse transcription- polymerase chain reaction (RT-PCR). The length of cGnRH-II cDNA1 and cDNA2 was 622 and 578 base pairs (bp), respectively. The cGnRH-II pre-cursors encoded by two cDNAs consisted of 86 amino acids, including a signal peptide, cGnRH-II decapeptide and a GnRH-associated peptide (GAP) linked by a Gly-Lys-Arg proteolytic site. The results of intron trapping and Southern blot showed that two differing cGnRH-II genes in common carp genome were further identified, and that two genes might exist as a single copy. The multi-gene coding of common carp cGnRH-II gene offered novel evidence for gene duplica-tion hypothesis. Using semi-quantitative RT-PCR, expression and relative expression levels of cGnRH-II genes were detected in five dissected brain regions, pituitary and gonad of common carp. With the exception of no mRNA2 in ovary, two cGnRH-II genes could be expressed in all the detected tissues. However, expression levels showed an apparent difference in different brain regions, pituitary and gonad. According to the expression characterization of cGnRH-II genes in brain areas, it was presumed that cGnRH-II might mainly work as the neurotransmitter and neuromodulator and also operate in the regulation for the GnRH releasing. Then, the ex-pression of cGnRH-II genes in pituitary and gonad suggested that cGnRH-II might act as the autocrine or paracrine regulator.
文摘To test the effectiveness of introducing live zooplankton against direct manuring in ornamental fish ponds upon their survival and production, larvae of koi carp, Cyprinus carpio L., were cultured for 11 weeks in earthen ponds maintained according to four management regimes： （1） live zooplankton fed to carp larvae （LF）; （2） direct fertilization with poultry manure （PM）; （3） direct fertilization with cowdung （CD）; and （4） a control treatment （C）. There were three replicates for each treatment. The growth of heterotrophic bacteria and pathogenic microorganisms like Aeromonas sp. and Pseudomonas sp. were also examined in response to pond management. Values of dissolved oxygen were significantly higher （P〈0.05） in the water of LF ponds, compared to other treatments, while the PM and CD treatments recorded were significantly higher （P〈0.05） values of PO4 - P, NH4 - N, NO3 - N, NO2 - N, specific conductivity, alkalinity, and BOD, compared to the LF and C treatments. The percentages of organic carbon and total nitrogen in the bottom sediments were higher in the PM and CD treatments compared to LF （P〈0.05）. Average counts of heterotrophic bacteria in the water of PM and CD ponds were significantly higher than other treatments （P〈0.05）. The development of Aeromonas sp. and Pseudomonas sp. were significantly higher （P〈0.05） in the PM and CD treatments. Weight gain of koi carp stocked in LF was significantly higher （P〈0.05） than that of fish in the other treatments. There was a significant difference in the survival rate of koi carp among the treatments ranging from 67.21% in C to 90.11% in LF. The results suggest that raising koi carp larvae in ponds and feeding them exogenously with zooplankton would support high rates of survival and production through maintenance of better water quality and greater abundance of zooplankton in the system. Significantly lower abundance of Aeromonas sp. and Pseudomonas sp. in the LF treatment considerably lowered any possibility of occurrence of bacterial disease.
