胭脂鱼幼鱼适宜蛋白水平及中草药对其非特异性免疫机能的影响
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摘要
本文以胭脂鱼(Myxocyprinus asiaticus)幼鱼为试验对象,研究涉及营养学和免疫学两部分,第一部分探讨了胭脂鱼幼鱼饲料中适宜的蛋白质需求量,第二部分探讨了3种中草药提取物的组方(包括蜂胶配淫羊藿、黄芪配甘草、陈皮)作为免疫增强剂对胭脂鱼幼鱼非特异性免疫力和抗病力的影响。主要研究结果如下:
(1)饲料蛋白水平对胭脂鱼幼鱼生长和体组成的影响:以体重(13.5±1.1)g的胭脂鱼为试验对象,以白鱼粉作为主要蛋白源,按6个蛋白质水平梯度(30%、34%、38%、42%、46%、50%)设计等能的饲料,进行8周的饲养试验,探讨胭脂鱼幼鱼适宜的蛋白质需求量。结果显示,饲料蛋白质水平对增重率(WG)和特定生长率(SGR)的影响基本均呈先升后降的变化趋势,WG和SGR最大值均出现在蛋白质含量46%的饲料组,分别为(187.70±8.01)%和(1.89±0.11)%/天,与蛋白质含量42%及以下的饲料组比较有显著性差异(P＜0.05),与蛋白质含量50%的饲料组间没有显著差异;根据折线模型计算出折点对应SGR的值为1.87%/天,对应的饲料蛋白质含量为46.5%;各组成活率(Survival)变化范围在71%～90%,不同饲料蛋白质含量对成活率无明显影响;随饲料蛋白质含量的增加,饲料系数(FCR)大体呈下降的趋势,最小值出现在46%饲料组;蛋白质效率(PER)和蛋白质利用率(PPV)均呈现先升高后降低的趋势,最大值均出现在46%饲料组;鱼体和肌肉粗蛋白的含量均随饲料蛋白质水平的升高而呈显著增加(P＜0.05);随饲料蛋白质含量的增加,肥满度(CF)、肝体指数(HSI)、脏体指数(VSI)均呈大体升高的趋势。结果表明,本试验条件下,胭脂鱼幼鱼饲料适宜蛋白质含量为46.5%。
(2)蜂胶配淫羊藿对胭脂鱼幼鱼非特异性免疫机能和抗病力的影响:醇提法制备中草药蜂胶提取物(总黄酮含量约37.1%)和淫羊藿提取物(总黄酮含量约42.6%),二者按生药重量比3:1合用,按质量分数0%(对照组)、0.1%、0.5%和1.0%添加于基础饲料成鳗粉中,饲喂胭脂鱼幼鱼,分别于第10 d、20 d、30 d、40 d、50 d采样,不连续密度梯度分离法制备头。肾巨噬细胞,检测细胞免疫指标:采用中性红吞噬试验检测吞噬活性,流式细胞术检测呼吸爆发活力;取血清,检测体液免疫指标:利用溶壁微球菌(Micrococcus lysoleikticus)检测溶菌酶活性,利用嗜水气单胞菌(Aeromonas hydrophila)检测杀菌活性,利用兔血红细胞检测替代补体溶血活性(ACH_(50))。结果显示,蜂胶配淫羊藿0.5%剂量组在第40 d和第50 d、1.0%剂量组在第30 d的头肾巨噬细胞吞噬活性均显著高于对照组(P＜0.05);0.5%剂量组在第30 d和40 d的头肾巨噬细胞呼吸爆发活力与对照组比较均有显著性升高(P＜0.05);0.1%剂量组在第40 d和第50 d、0.5%剂量组在第20 d和第40 d、1.0%剂量组在第10 d的血清溶菌酶活性均显著高于对照组(P＜0.05);第40 d,0.5%剂量组的杀菌活性与对照组比较有显著升高(P＜0.05);蜂胶配淫羊藿对ACHso则无明显影响。饲喂50 d后,各组随机取20尾鱼,用A.hydrophila腹腔注射进行攻毒试验,探讨蜂胶配淫羊藿对胭脂鱼幼鱼抗病力的影响,结果显示1周内对照组的累积死亡率为75%,1.0%和0.1%剂量组的累积死亡率分别为45%和55%,0.5%剂量组的累积死亡率40%则显著低于对照组(P＜0.05)。表明蜂胶配淫羊藿可开发为一种新型的免疫增强剂,饲料中适宜添加剂量为0.5%。
(3)黄芪配甘草对胭脂鱼幼鱼非特异性免疫机能和抗病力的影响:醇提法制备黄芪提取物(黄芪多糖含量约为38.4%)和甘草提取物(甘草酸含量约为31.2%,总黄酮约为20.5%),二者按生药重量比10:6合用,按质量分数0%(对照组)、0.1%、0.5%和1.0%添加于成鳗粉中,饲喂胭脂鱼幼鱼,分别于第10 d、20 d、30 d、40 d、50 d各取样一次,分离头肾巨噬细胞,检测吞噬活性和呼吸爆发活力等细胞免疫指标;分离血清,检测溶菌酶活性、杀菌活性和替代补体溶血活性等体液免疫指标。结果显示,1.0%剂量组在第30 d和第40 d、0.5%剂量组在第40 d以及0.1%剂量组在第50 d的头肾巨噬细胞吞噬活性均显著高于对照组(P＜0.05);0.5%剂量组在第20 d和第40 d、1.0%剂量组在第40 d和第50 d的头肾巨噬细胞呼吸爆发活力与对照组比较均有显著性升高(P＜0.05);1.0%剂量组在第20 d和第50 d、0.5%剂量组在第30 d以及0.1%剂量组在第40 d的血清溶菌酶活性均有明显增加,与对照组比较差异显著(P＜0.05);1.0%剂量组在第30 d和第40 d、0.5%剂量组在第40 d和第50 d的杀菌百分率均有明显升高,与对照组比较有显著性差异(P＜0.05);1.0%剂量组在第40和50 d、0.5%剂量组在第30 d以及0.1%剂量组在第50 d的ACH_(50)与对照组比较均有明显升高(P＜0.05)。饲喂50 d后,各组随机取20尾鱼,用A.hydrophila腹腔注射进行攻毒试验,探讨黄芪配甘草对胭脂鱼幼鱼抗病力的影响,结果显示1周内0.5%和0.1%剂量组的累积死亡率分别为45%和60%,1.0%剂量组的累积死亡率35%则显著低于对照组(P＜0.05)。结果表明,黄芪配甘草可开发为一种新型的免疫增强剂,对胭脂鱼幼鱼免疫功能的提高表现出剂量一效应关系,适宜添加剂量为1.0%。
(4)陈皮对胭脂鱼幼鱼非特异性免疫机能和抗病力的影响:醇提法制备陈皮提取物(总黄酮含量约为3.3%,橙皮苷含量约为1.6%),按质量分数0%(对照组)、0.1%、0.5%和1.0%添加于成鳗粉中,饲喂胭脂鱼幼鱼,分别于第10 d、20 d、30 d、40 d、50 d各取样一次,分离头肾巨噬细胞,检测吞噬活性和呼吸爆发活力;分离血清,检测溶菌酶活性、杀菌活性和替代补体溶血活性。结果显示,第50 d时,陈皮提取物1.0%剂量组的头肾巨噬细胞吞噬活性显著高于对照组(P＜0.05);0.5%剂量组在第30 d、1.0%剂量组在第40 d的头肾巨噬细胞呼吸爆发活力与对照组比较均有显著性升高(P＜0.05);第20 d,1.0%剂量组的血清溶菌酶活性比对照组有明显增加(P＜0.05);陈皮提取物对血清杀菌活性和ACHso则无明显影响。饲喂50 d后,各组随机取20尾鱼,用A.hydrophila腹腔注射进行攻毒试验,探讨陈皮对胭脂鱼幼鱼抗病力的影响,结果显示1周内1.0%、0.5%、0.1%剂量组的累积死亡率分别为50%、55%、65%,与对照组比较差异并不显著(P＞0.05)。结果表明陈皮提取物能够在一定程度上增强胭脂鱼幼鱼的非特异性免疫力,关于陈皮中单一成分橙皮苷对胭脂鱼幼鱼非特异性免疫力和抗病力的影响尚需进一步研究。
(5)中药提取物的体外免疫试验:以胭脂鱼幼鱼的头肾巨噬细胞为研究对象,探讨两种中药提取物组方(蜂胶配淫羊藿、黄芪配甘草)对离体培养细胞免疫活性的影响。不连续密度梯度分离法制备头肾巨噬细胞,纯化,接种到96孔培养板,加入终浓度依次为0、25、50、100、200、400μg/mL的中药提取物溶液,共培养24 h,NBT法检测头肾巨噬细胞的呼吸爆发活力。结果显示蜂胶配淫羊藿在浓度25、50、100μg/mL时,黄芪配甘草浓度为100、200μg/mL时,呼吸爆发活力值与空白对照组比较均有显著差异(P＜0.05)。这一结果从体外验证了2种中药提取物组方在胭脂鱼幼鱼养殖试验中的免疫增强效果。
This study was conducted on juvenile Chinese sucker, Myxocyprinus asiaticus. In the first part, the dietary protein requirement of juvenile Chinese sucker was determined. In the second part, the effects of three traditional Chinese medicines (TCM) formulated from propolis and Herba Epimedii extracts, Radix Astragali and Glycyrrhiza uralensis extracts, Tangerine Peel extract respectively, on non-specific immune response of Chinese sucker were investigated. The results were as follows.
(1) Growth performance and proximate composition of juvenile Chinese sucker fed the experimental diets containing different protein levels: Six fishmeal-based experimental diets containing various crude protein levels including 30%, 34%, 38%, 42%, 46% and 50% were fed to triplicate groups of 20 fish each (initial weight 13.5±1.1 g) for 56 days. Weight gain (WG) and specific growth rate (SGR) increased with increasing dietary protein level up to 46% (P < 0.05). Based on broken-line regression analysis of SGR against dietary protein levels the optimal dietary protein requirement for juvenile Chinese sucker was estimated to be close to 46.5%. Survival was not affected by dietary protein level (overall survival 71 - 90%). Feed conversion rate (FCR) generally showed a decline with higher protein levels (from 1.62 in 30% protein group to 1.13 in 50% protein group). Protein efficiency ratio (PER) showed gradual improvements with increasing dietary protein up to 46%. A similar trend was found for protein productive value (PPV). Among the proximate compositions of the whole body and muscle, crude protein contents all increased significantly with increasing dietary protein levels (P < 0.05). Conditon factor (CF), hepatosomatic index (HSI) and viscerosomatic index (VSI) generally showed an increase with higher protein levels. It was estimated that the optimal dietary protein requirement for juvenile Chinese sucker was close to 46.5%.
(2) Effects of combination of propolis and Herba Epimedii extracts on the non-specific immune response and disease resistance of Chinese sucker: Propolis extract (Total flavone content: 37.1%) and Herba Epimedii extract (Total flavone content: 42.6%) was extracted using alcohol and mixed at the ratio of 3:1 (w/w). Fish were fed diets containing 0 (control), 0.1%, 0.5% or 1.0% TCM extracts for 50 days. Head-kidney and blood samples were collected at 10 d, 20 d, 30 d, 40 d and 50 d, respectively. Macrophages in head-kidney were separated by discontinuous density gradient centrifugation. The phagocytic activity of macrophage was measured using neutral red. The respiratory burst activity of macrophage was measured by flow cytometry (FCM). The lysozyme, bactericidal and natural haemolytic complement (ACH_(50)) activities in blood serum were measured using Micrococcus lysoleikticus, Aeromonas hydrophila and RaRBC, respectively. Results of this study showed that feeding the fish with 0.5% TCM extracts significantly increased the phagocytic activity after 40 days (P < 0.05). The enhanced phagocytic activity remained as compared to the control until the end of the experiment. Fish fed with 1.0% TCM extracts showed enhanced phagocytic activity on the 30 days compared to the control. The respiratory burst activity of macrophage showed a statistically significant increase on the 30 and 40 days in 0.5% dose group compared to the control (P < 0.05). A significant increasing of lysozyme activity was found after 10 days in group fed with 1.0% TCM extracts compared to the control (P < 0.05). Significant differences were also measured in 0.5% dose group on the 20 and 40 days and 0.1% dose group on the 40 days and the end of the experiment. Feeding the fish with 0.5% TCM extracts significantly increased the bactericidal activity after 40 days (P < 0.05). There were no significant changes in ACH_(50) during feeding. After 50 days of feeding fish were challenged with A. hydrophila and cumulative mortality was registered during one week. A cumulative mortality over the experimental period in the control group was 75%. All dosage of treated groups showed reduced mortality compared to the control. Cumulative mortalities in fish fed with 0.1% and 1.0% TCM extracts reached 45% and 55%, respectively. Feed containing 0.5% TCM extracts was the most effective with the mortality of the fish significantly reduced by 35% compared to the control. The results indicate that propolis and Herba Epimedii extracts in combination enhances the non-specific immune response and disease resistance of Chinese sucker against A. hydrophila. The optimal dosage added to feed was 0.5% (w/w).
(3) Effects of combination of Radix Astragali and Glycyrrhiza uralensis extracts on the non-specific immune response and disease resistance of Chinese sucker: Radix Astragali extract (Astragalus polysaccharides content: 38.4%) and Glycyrrhiza uralensis extract (Glycyrrhizic acid content: 31.2%; total flavone content: 20.5%) was extracted using alcohol and mixed at the ratio of 10:6 (w/w). Fish were fed diets containing 0 (control), 0.1%, 0.5% or 1.0% TCM extracts for 50 days. Head-kidney and blood samples were collected at 10 d, 20 d, 30 d, 40 d and 50 d, respectively. Macrophages in head-kidney were separated by discontinuous density gradient centrifugation. The phagocytic activity of macrophage was measured using neutral red. The respiratory burst activity of macrophage was measured by FCM. The lysozyme, bactericidal and ACH_(50) activities in blood serum were measured using M. lysoleikticus, A. hydrophila and RaRBC, respectively. Results of this study showed that the phagocytic activity of macrophage showed a statistically significant increase on the 30 and 40 days in 1.0% dose group, 40 days in 0.5% dose group and 50 days in 0.1% dose group compared to the control (P < 0.05). Feeding the fish with 1.0% TCM extracts significantly increased the respiratory burst activity of macrophage after 40 days. The enhanced activity remained as compared to the control until the end of the experiment (P < 0.05). A significant increasing was also found on the 20 and 40 days in 0.5% dose group compared to the control. Significant differences of lysozyme activity was measured in 1.0% dose group on the 20 and 50 days, 0.5% dose group on the 30 days and 0.1% dose group on the 40 days compared to the control (P < 0.05). Feeding the fish with 0.5% TCM extracts significantly increased the bactericidal activity after 40 days until the end of the experiment (P < 0.05). A significant increasing was also found on the 30 and 40 days in group fed with 1.0% TCM extracts compared to the control. The ACH_(50) activity showed a statistically significant increase on the 40 and 50 days in 1.0% dose group, 30 days in 0.5% dose group and 50 days in 0.1% dose group compared to the control (P < 0.05). After 50 days of feeding fish were challenged with A. hydrophila and cumulative mortality was registered during one week. A cumulative mortality over the experimental period in the control group was 75%. All dosage of treated groups showed reduced mortality compared to the control. Cumulative mortalities in fish fed with 0.1% and 0.5% TCM extracts reached 60% and 45%, respectively. Feed containing 1.0% TCM extracts was the most effective with the mortality of the fish significantly reduced by 40% compared to the control (P < 0.05). The results indicate that Radix Astragali and Glycyrrhiza uralensis extracts in combination enhances the non-specific immune response and disease resistance of Chinese sucker against A. hydrophila. The optimal dosage added to feed was 1.0% (w/w).
(4) Effects of Tangerine Peel extract on the non-specific immune response and disease resistance of Chinese sucker: Tangerine Peel extract (Total flavone content: 3.3%; Hesperidin content: 1.6%) was extracted using alcohol. Fish were fed diets containing 0 (control), 0.1%, 0.5% or 1.0% Tangerine Peel extract for 50 days. Head-kidney and blood samples were collected at 10 d, 20 d, 30 d, 40 d and 50 d, respectively. Macrophages in head-kidney were separated by discontinuous density gradient centrifugation. The phagocytic activity of macrophage was measured using neutral red. The respiratory burst activity of macrophage was measured by FCM. The lysozyme, bactericidal and ACH_(50) activities in blood serum were measured using M. lysoleikticus, A. hydrophila and RaRBC, respectively. Results of this study showed that the phagocytic activity of macrophage had a statistically significant increase on the 50 days in 1.0% dose group compared to the control (P < 0.05). Significant differences of respiratory burst activity was measured in 1.0% dose group on the 40 days and 0.5% dose group on the 30 days compared to the control (P < 0.05). Feeding the fish with 1.0% Tangerine Peel extract significantly increased the lysozyme activity in blood serum after 20 days (P < 0.05). There were no significant changes in bactericidal and ACH_(50) activities comparing to the control during the whole experiment. After 50 days of feeding fish were challenged with A. hydrophila and cumulative mortality was registered during one week. A cumulative mortality over the experimental period in the control group was 75%. Cumulative mortalities in fish fed with 0.1%, 0.5% and 1.0% Tangerine Peel extract reached 65%, 55% and 50%, respectively. The results indicate that Tangerine Peel extract enhances the non-specific immune response and disease resistance of Chinese sucker against A. hydrophila.
(5) Effects of TCM extracts on respiratory burst activity of head-kidney macrophage of Chinese sucker in vitro: Macrophages in head-kidney of Chinese sucker were separated and purified by discontinuous density gradient centrifugation. Then head-kidney macrophages were incubated in a 96-well plate with different doses of TCM extracts (combination of propolis and Herba Epimedii extracts, combination of Radix Astragali and Glycyrrhiza uralensis extracts), 0, 25, 50, 100, 200 and 400μg/mL, respectively, for 24 h. The respiratory burst activity of macrophage was measured by nitroblue tetrazolium (NBT). Significant differences of respiratory burst activity was found in combination of propolis and Herba Epimedii extracts with doses of 25, 50 and 100μg/mL and combination of Radix Astragali and Glycyrrhiza uralensis extracts with doses of 100 and 200μg/mL compared to the control (P < 0.05). The results indicate that combination of propolis and Herba Epimedii extracts and combination of Radix Astragali and Glycyrrhiza uralensis extracts could modulate the immune response of Chinese sucker in vitro.
(1)饲料蛋白水平对胭脂鱼幼鱼生长和体组成的影响:以体重(13.5±1.1)g的胭脂鱼为试验对象,以白鱼粉作为主要蛋白源,按6个蛋白质水平梯度(30%、34%、38%、42%、46%、50%)设计等能的饲料,进行8周的饲养试验,探讨胭脂鱼幼鱼适宜的蛋白质需求量。结果显示,饲料蛋白质水平对增重率(WG)和特定生长率(SGR)的影响基本均呈先升后降的变化趋势,WG和SGR最大值均出现在蛋白质含量46%的饲料组,分别为(187.70±8.01)%和(1.89±0.11)%/天,与蛋白质含量42%及以下的饲料组比较有显著性差异(P＜0.05),与蛋白质含量50%的饲料组间没有显著差异;根据折线模型计算出折点对应SGR的值为1.87%/天,对应的饲料蛋白质含量为46.5%;各组成活率(Survival)变化范围在71%～90%,不同饲料蛋白质含量对成活率无明显影响;随饲料蛋白质含量的增加,饲料系数(FCR)大体呈下降的趋势,最小值出现在46%饲料组;蛋白质效率(PER)和蛋白质利用率(PPV)均呈现先升高后降低的趋势,最大值均出现在46%饲料组;鱼体和肌肉粗蛋白的含量均随饲料蛋白质水平的升高而呈显著增加(P＜0.05);随饲料蛋白质含量的增加,肥满度(CF)、肝体指数(HSI)、脏体指数(VSI)均呈大体升高的趋势。结果表明,本试验条件下,胭脂鱼幼鱼饲料适宜蛋白质含量为46.5%。
(2)蜂胶配淫羊藿对胭脂鱼幼鱼非特异性免疫机能和抗病力的影响:醇提法制备中草药蜂胶提取物(总黄酮含量约37.1%)和淫羊藿提取物(总黄酮含量约42.6%),二者按生药重量比3:1合用,按质量分数0%(对照组)、0.1%、0.5%和1.0%添加于基础饲料成鳗粉中,饲喂胭脂鱼幼鱼,分别于第10 d、20 d、30 d、40 d、50 d采样,不连续密度梯度分离法制备头。肾巨噬细胞,检测细胞免疫指标:采用中性红吞噬试验检测吞噬活性,流式细胞术检测呼吸爆发活力;取血清,检测体液免疫指标:利用溶壁微球菌(Micrococcus lysoleikticus)检测溶菌酶活性,利用嗜水气单胞菌(Aeromonas hydrophila)检测杀菌活性,利用兔血红细胞检测替代补体溶血活性(ACH_(50))。结果显示,蜂胶配淫羊藿0.5%剂量组在第40 d和第50 d、1.0%剂量组在第30 d的头肾巨噬细胞吞噬活性均显著高于对照组(P＜0.05);0.5%剂量组在第30 d和40 d的头肾巨噬细胞呼吸爆发活力与对照组比较均有显著性升高(P＜0.05);0.1%剂量组在第40 d和第50 d、0.5%剂量组在第20 d和第40 d、1.0%剂量组在第10 d的血清溶菌酶活性均显著高于对照组(P＜0.05);第40 d,0.5%剂量组的杀菌活性与对照组比较有显著升高(P＜0.05);蜂胶配淫羊藿对ACHso则无明显影响。饲喂50 d后,各组随机取20尾鱼,用A.hydrophila腹腔注射进行攻毒试验,探讨蜂胶配淫羊藿对胭脂鱼幼鱼抗病力的影响,结果显示1周内对照组的累积死亡率为75%,1.0%和0.1%剂量组的累积死亡率分别为45%和55%,0.5%剂量组的累积死亡率40%则显著低于对照组(P＜0.05)。表明蜂胶配淫羊藿可开发为一种新型的免疫增强剂,饲料中适宜添加剂量为0.5%。
(3)黄芪配甘草对胭脂鱼幼鱼非特异性免疫机能和抗病力的影响:醇提法制备黄芪提取物(黄芪多糖含量约为38.4%)和甘草提取物(甘草酸含量约为31.2%,总黄酮约为20.5%),二者按生药重量比10:6合用,按质量分数0%(对照组)、0.1%、0.5%和1.0%添加于成鳗粉中,饲喂胭脂鱼幼鱼,分别于第10 d、20 d、30 d、40 d、50 d各取样一次,分离头肾巨噬细胞,检测吞噬活性和呼吸爆发活力等细胞免疫指标;分离血清,检测溶菌酶活性、杀菌活性和替代补体溶血活性等体液免疫指标。结果显示,1.0%剂量组在第30 d和第40 d、0.5%剂量组在第40 d以及0.1%剂量组在第50 d的头肾巨噬细胞吞噬活性均显著高于对照组(P＜0.05);0.5%剂量组在第20 d和第40 d、1.0%剂量组在第40 d和第50 d的头肾巨噬细胞呼吸爆发活力与对照组比较均有显著性升高(P＜0.05);1.0%剂量组在第20 d和第50 d、0.5%剂量组在第30 d以及0.1%剂量组在第40 d的血清溶菌酶活性均有明显增加,与对照组比较差异显著(P＜0.05);1.0%剂量组在第30 d和第40 d、0.5%剂量组在第40 d和第50 d的杀菌百分率均有明显升高,与对照组比较有显著性差异(P＜0.05);1.0%剂量组在第40和50 d、0.5%剂量组在第30 d以及0.1%剂量组在第50 d的ACH_(50)与对照组比较均有明显升高(P＜0.05)。饲喂50 d后,各组随机取20尾鱼,用A.hydrophila腹腔注射进行攻毒试验,探讨黄芪配甘草对胭脂鱼幼鱼抗病力的影响,结果显示1周内0.5%和0.1%剂量组的累积死亡率分别为45%和60%,1.0%剂量组的累积死亡率35%则显著低于对照组(P＜0.05)。结果表明,黄芪配甘草可开发为一种新型的免疫增强剂,对胭脂鱼幼鱼免疫功能的提高表现出剂量一效应关系,适宜添加剂量为1.0%。
(4)陈皮对胭脂鱼幼鱼非特异性免疫机能和抗病力的影响:醇提法制备陈皮提取物(总黄酮含量约为3.3%,橙皮苷含量约为1.6%),按质量分数0%(对照组)、0.1%、0.5%和1.0%添加于成鳗粉中,饲喂胭脂鱼幼鱼,分别于第10 d、20 d、30 d、40 d、50 d各取样一次,分离头肾巨噬细胞,检测吞噬活性和呼吸爆发活力;分离血清,检测溶菌酶活性、杀菌活性和替代补体溶血活性。结果显示,第50 d时,陈皮提取物1.0%剂量组的头肾巨噬细胞吞噬活性显著高于对照组(P＜0.05);0.5%剂量组在第30 d、1.0%剂量组在第40 d的头肾巨噬细胞呼吸爆发活力与对照组比较均有显著性升高(P＜0.05);第20 d,1.0%剂量组的血清溶菌酶活性比对照组有明显增加(P＜0.05);陈皮提取物对血清杀菌活性和ACHso则无明显影响。饲喂50 d后,各组随机取20尾鱼,用A.hydrophila腹腔注射进行攻毒试验,探讨陈皮对胭脂鱼幼鱼抗病力的影响,结果显示1周内1.0%、0.5%、0.1%剂量组的累积死亡率分别为50%、55%、65%,与对照组比较差异并不显著(P＞0.05)。结果表明陈皮提取物能够在一定程度上增强胭脂鱼幼鱼的非特异性免疫力,关于陈皮中单一成分橙皮苷对胭脂鱼幼鱼非特异性免疫力和抗病力的影响尚需进一步研究。
(5)中药提取物的体外免疫试验:以胭脂鱼幼鱼的头肾巨噬细胞为研究对象,探讨两种中药提取物组方(蜂胶配淫羊藿、黄芪配甘草)对离体培养细胞免疫活性的影响。不连续密度梯度分离法制备头肾巨噬细胞,纯化,接种到96孔培养板,加入终浓度依次为0、25、50、100、200、400μg/mL的中药提取物溶液,共培养24 h,NBT法检测头肾巨噬细胞的呼吸爆发活力。结果显示蜂胶配淫羊藿在浓度25、50、100μg/mL时,黄芪配甘草浓度为100、200μg/mL时,呼吸爆发活力值与空白对照组比较均有显著差异(P＜0.05)。这一结果从体外验证了2种中药提取物组方在胭脂鱼幼鱼养殖试验中的免疫增强效果。
This study was conducted on juvenile Chinese sucker, Myxocyprinus asiaticus. In the first part, the dietary protein requirement of juvenile Chinese sucker was determined. In the second part, the effects of three traditional Chinese medicines (TCM) formulated from propolis and Herba Epimedii extracts, Radix Astragali and Glycyrrhiza uralensis extracts, Tangerine Peel extract respectively, on non-specific immune response of Chinese sucker were investigated. The results were as follows.
(1) Growth performance and proximate composition of juvenile Chinese sucker fed the experimental diets containing different protein levels: Six fishmeal-based experimental diets containing various crude protein levels including 30%, 34%, 38%, 42%, 46% and 50% were fed to triplicate groups of 20 fish each (initial weight 13.5±1.1 g) for 56 days. Weight gain (WG) and specific growth rate (SGR) increased with increasing dietary protein level up to 46% (P < 0.05). Based on broken-line regression analysis of SGR against dietary protein levels the optimal dietary protein requirement for juvenile Chinese sucker was estimated to be close to 46.5%. Survival was not affected by dietary protein level (overall survival 71 - 90%). Feed conversion rate (FCR) generally showed a decline with higher protein levels (from 1.62 in 30% protein group to 1.13 in 50% protein group). Protein efficiency ratio (PER) showed gradual improvements with increasing dietary protein up to 46%. A similar trend was found for protein productive value (PPV). Among the proximate compositions of the whole body and muscle, crude protein contents all increased significantly with increasing dietary protein levels (P < 0.05). Conditon factor (CF), hepatosomatic index (HSI) and viscerosomatic index (VSI) generally showed an increase with higher protein levels. It was estimated that the optimal dietary protein requirement for juvenile Chinese sucker was close to 46.5%.
(2) Effects of combination of propolis and Herba Epimedii extracts on the non-specific immune response and disease resistance of Chinese sucker: Propolis extract (Total flavone content: 37.1%) and Herba Epimedii extract (Total flavone content: 42.6%) was extracted using alcohol and mixed at the ratio of 3:1 (w/w). Fish were fed diets containing 0 (control), 0.1%, 0.5% or 1.0% TCM extracts for 50 days. Head-kidney and blood samples were collected at 10 d, 20 d, 30 d, 40 d and 50 d, respectively. Macrophages in head-kidney were separated by discontinuous density gradient centrifugation. The phagocytic activity of macrophage was measured using neutral red. The respiratory burst activity of macrophage was measured by flow cytometry (FCM). The lysozyme, bactericidal and natural haemolytic complement (ACH_(50)) activities in blood serum were measured using Micrococcus lysoleikticus, Aeromonas hydrophila and RaRBC, respectively. Results of this study showed that feeding the fish with 0.5% TCM extracts significantly increased the phagocytic activity after 40 days (P < 0.05). The enhanced phagocytic activity remained as compared to the control until the end of the experiment. Fish fed with 1.0% TCM extracts showed enhanced phagocytic activity on the 30 days compared to the control. The respiratory burst activity of macrophage showed a statistically significant increase on the 30 and 40 days in 0.5% dose group compared to the control (P < 0.05). A significant increasing of lysozyme activity was found after 10 days in group fed with 1.0% TCM extracts compared to the control (P < 0.05). Significant differences were also measured in 0.5% dose group on the 20 and 40 days and 0.1% dose group on the 40 days and the end of the experiment. Feeding the fish with 0.5% TCM extracts significantly increased the bactericidal activity after 40 days (P < 0.05). There were no significant changes in ACH_(50) during feeding. After 50 days of feeding fish were challenged with A. hydrophila and cumulative mortality was registered during one week. A cumulative mortality over the experimental period in the control group was 75%. All dosage of treated groups showed reduced mortality compared to the control. Cumulative mortalities in fish fed with 0.1% and 1.0% TCM extracts reached 45% and 55%, respectively. Feed containing 0.5% TCM extracts was the most effective with the mortality of the fish significantly reduced by 35% compared to the control. The results indicate that propolis and Herba Epimedii extracts in combination enhances the non-specific immune response and disease resistance of Chinese sucker against A. hydrophila. The optimal dosage added to feed was 0.5% (w/w).
(3) Effects of combination of Radix Astragali and Glycyrrhiza uralensis extracts on the non-specific immune response and disease resistance of Chinese sucker: Radix Astragali extract (Astragalus polysaccharides content: 38.4%) and Glycyrrhiza uralensis extract (Glycyrrhizic acid content: 31.2%; total flavone content: 20.5%) was extracted using alcohol and mixed at the ratio of 10:6 (w/w). Fish were fed diets containing 0 (control), 0.1%, 0.5% or 1.0% TCM extracts for 50 days. Head-kidney and blood samples were collected at 10 d, 20 d, 30 d, 40 d and 50 d, respectively. Macrophages in head-kidney were separated by discontinuous density gradient centrifugation. The phagocytic activity of macrophage was measured using neutral red. The respiratory burst activity of macrophage was measured by FCM. The lysozyme, bactericidal and ACH_(50) activities in blood serum were measured using M. lysoleikticus, A. hydrophila and RaRBC, respectively. Results of this study showed that the phagocytic activity of macrophage showed a statistically significant increase on the 30 and 40 days in 1.0% dose group, 40 days in 0.5% dose group and 50 days in 0.1% dose group compared to the control (P < 0.05). Feeding the fish with 1.0% TCM extracts significantly increased the respiratory burst activity of macrophage after 40 days. The enhanced activity remained as compared to the control until the end of the experiment (P < 0.05). A significant increasing was also found on the 20 and 40 days in 0.5% dose group compared to the control. Significant differences of lysozyme activity was measured in 1.0% dose group on the 20 and 50 days, 0.5% dose group on the 30 days and 0.1% dose group on the 40 days compared to the control (P < 0.05). Feeding the fish with 0.5% TCM extracts significantly increased the bactericidal activity after 40 days until the end of the experiment (P < 0.05). A significant increasing was also found on the 30 and 40 days in group fed with 1.0% TCM extracts compared to the control. The ACH_(50) activity showed a statistically significant increase on the 40 and 50 days in 1.0% dose group, 30 days in 0.5% dose group and 50 days in 0.1% dose group compared to the control (P < 0.05). After 50 days of feeding fish were challenged with A. hydrophila and cumulative mortality was registered during one week. A cumulative mortality over the experimental period in the control group was 75%. All dosage of treated groups showed reduced mortality compared to the control. Cumulative mortalities in fish fed with 0.1% and 0.5% TCM extracts reached 60% and 45%, respectively. Feed containing 1.0% TCM extracts was the most effective with the mortality of the fish significantly reduced by 40% compared to the control (P < 0.05). The results indicate that Radix Astragali and Glycyrrhiza uralensis extracts in combination enhances the non-specific immune response and disease resistance of Chinese sucker against A. hydrophila. The optimal dosage added to feed was 1.0% (w/w).
(4) Effects of Tangerine Peel extract on the non-specific immune response and disease resistance of Chinese sucker: Tangerine Peel extract (Total flavone content: 3.3%; Hesperidin content: 1.6%) was extracted using alcohol. Fish were fed diets containing 0 (control), 0.1%, 0.5% or 1.0% Tangerine Peel extract for 50 days. Head-kidney and blood samples were collected at 10 d, 20 d, 30 d, 40 d and 50 d, respectively. Macrophages in head-kidney were separated by discontinuous density gradient centrifugation. The phagocytic activity of macrophage was measured using neutral red. The respiratory burst activity of macrophage was measured by FCM. The lysozyme, bactericidal and ACH_(50) activities in blood serum were measured using M. lysoleikticus, A. hydrophila and RaRBC, respectively. Results of this study showed that the phagocytic activity of macrophage had a statistically significant increase on the 50 days in 1.0% dose group compared to the control (P < 0.05). Significant differences of respiratory burst activity was measured in 1.0% dose group on the 40 days and 0.5% dose group on the 30 days compared to the control (P < 0.05). Feeding the fish with 1.0% Tangerine Peel extract significantly increased the lysozyme activity in blood serum after 20 days (P < 0.05). There were no significant changes in bactericidal and ACH_(50) activities comparing to the control during the whole experiment. After 50 days of feeding fish were challenged with A. hydrophila and cumulative mortality was registered during one week. A cumulative mortality over the experimental period in the control group was 75%. Cumulative mortalities in fish fed with 0.1%, 0.5% and 1.0% Tangerine Peel extract reached 65%, 55% and 50%, respectively. The results indicate that Tangerine Peel extract enhances the non-specific immune response and disease resistance of Chinese sucker against A. hydrophila.
(5) Effects of TCM extracts on respiratory burst activity of head-kidney macrophage of Chinese sucker in vitro: Macrophages in head-kidney of Chinese sucker were separated and purified by discontinuous density gradient centrifugation. Then head-kidney macrophages were incubated in a 96-well plate with different doses of TCM extracts (combination of propolis and Herba Epimedii extracts, combination of Radix Astragali and Glycyrrhiza uralensis extracts), 0, 25, 50, 100, 200 and 400μg/mL, respectively, for 24 h. The respiratory burst activity of macrophage was measured by nitroblue tetrazolium (NBT). Significant differences of respiratory burst activity was found in combination of propolis and Herba Epimedii extracts with doses of 25, 50 and 100μg/mL and combination of Radix Astragali and Glycyrrhiza uralensis extracts with doses of 100 and 200μg/mL compared to the control (P < 0.05). The results indicate that combination of propolis and Herba Epimedii extracts and combination of Radix Astragali and Glycyrrhiza uralensis extracts could modulate the immune response of Chinese sucker in vitro.
引文
艾春汉,邹金虎,.喻运珍,艾桃山.6种中药对鲫非特异性免疫效果的影响.淡水渔业,2009,39(1):76-79
蔡春芳,宋学宏,潘兴法,成文靖,浦佩,朱素琴.几种抗病促生长剂对银鲫生长和免疫力的影响.水利渔业,2002,22(2):20-22
曹丽萍,丁炜东,张柳,Galina J,徐跑,殷国俊.香菇多糖和黄芪多糖对鲤免疫细胞的活性和IL-1β体外诱生表达的影响.水产学报,2008a,32(4):628-635
曹丽萍,丁炜东,张柳,Galina J,殷国俊.2种多糖对鲤鱼离体培养免疫细胞活性的影响.安徽农业大学学报,2008b,35(2):219-223
曹振杰,曲世科,丛日祥,马俊岭,王志忠,刘汉华,张家国,孙鹤田,轩子群.免疫多糖对草鱼免疫功能的影响.齐鲁渔业,1999,16(3):43-44
陈昌福,罗宇良,蔡冰.饲养水温对草鱼溶菌酶活性的影响.中国水产科学,1996,3(3):24-30
陈超然,陈晓辉,陈昌福.口服甘草素对中华鳖稚鳖抗嗜水气单胞菌感染的作用.华中农业大学学报,2000,19(6):577-580
陈家长,简纪常.水体中NH3-N对建鲤非特异性免疫功能的影响.湛江海洋大学学报,2000,20(3):13-16
陈金生,刘家寿.不同饲料养殖一龄胭脂鱼种生长试验.水利渔业,1992,4:9-11,14
陈忻,黄耀威,赖兴华,杨桂芳.鱼类饲料甘草添加剂的研究.佛山科学技术学院学报(自然科学版),1999,17(3):36-38
陈孝煊,吴志新,殷居易,李莉.大黄、穿心莲、板蓝根和金银花对异育银鲫免疫机能的影响.中国水产科学,2003,10(1):36-39
陈孝煊,吴志新,张厚梅.大黄与黄连对二种淡水虾血细胞吞噬活性的影响.水生生物学报,2002,26(2):201-204
陈兆.胭脂鱼的生物学特性及饲养技术.渔业致富指南,2005,22:40-43
陈振昆,丁光.陈皮对草鱼诱食作用的研究.云南农业大学学报,1996,11(1):35-37
邓中群,余志堂,赵燕.三峡水利枢纽对长江白鲟和胭脂鱼影响的评价及资源保护的研究.见:长江三峡工程对生志与环境影响及其对策研究论文集.北京:科学出版社,1987,42-52
丁光,陈振昆,李梅,李国治.饲料添加剂陈皮对草鱼淋巴细胞转化率影响的研究.云南农业大学学报,2003,18(4):401-403
丁光,陈振昆,李文贵,杨贵树,许春艳.陈皮对草鱼细胞免疫影响的研究.云南农业大学学报,2004,19(6):727-730
丁光,陈振昆,徐宝明.在饲料中添加柑橘或甜橙外果皮对鲤鱼诱食作用的比较研究.水产科学,1999,18(1):16-19
杜爱芳,叶均安,于涟.复方大蒜油添加剂对中国对虾免疫机能的增强作用.浙江农业大学学报,1997,23(3):317-320
付自东,李静,岳碧松,宋昭彬.用荧光物质浸泡标记胭脂鱼仔、稚鱼耳石.动物学杂志,2005,40(4):60-65
龚宏伟,蔡春芳,阙林林.长江胭脂鱼开口饵料的研究.水产科学,2005,24(11):7-9
郭宝林,余竞光,肖培根.淫羊霍化学成分的研究.中国中药杂志,1996,21(5):294-296
郭文婷.两种免疫增强剂对牙鲆和凡纳滨对虾免疫因子影响的研究.f硕士学位论文].青岛:中国海洋大学,2005
郭永军,刑克智,陈成勋,杨羽兴,朱国霞.几种中草药对鲤鱼诱食效果的研究.天津农学院学报,2005,12(3):1-5
郝炳华,陶明,何立端,薛晚利,李选社,孟晓军,张京田.淫羊藿总黄酮对荷瘤小鼠细胞免疫及红细胞免疫功能的影响.西安医科大学学报,1999,20(3):323-325
胡兵,刘军,侯永清,李惠,陈爱敬,辜玲芳.黄芪多糖对异育银鲫非特异性免疫力的影响.水利渔业,2008,28(3):108-111
湖南省水产科学研究所.湖南鱼类志.长沙:湖南科学技术出版社,1980,24-25.
胡隐昌.胭脂鱼的主要生物学.珠江水产,2001,2:33-35
胡元亮,刘家国,宋大鲁,许志刚.淫蜂佐剂影响动物免疫功能的实验研究.畜 牧兽医学报,2000,31(2):155-160
胡元亮,吕殿红,刘家国,宋大鲁.淫羊藿.蜂胶佐剂对雏鸡和小白鼠腹腔巨噬细胞活性的影响.中国兽医科技,1999,29(5):8-10
黄克安,朱木洛,吴杏清.莨菪类药物防治草鱼出血病试验.鱼病简讯,1985,(1):20-23
黄小丽,邓永强.中草药免疫增强剂在水产上的开发应用.内陆水产,2004,8:24-26
吉红,李海梅,周继术,刘兰锋.4种植物粗提物对金鱼诱食效果的初步研究.水产科学,2008,27(2):67-70
简纪常,吴灶和.中草药对建鲤非特异性免疫功能的影响.大连水产学院学报,2002,17(2):114-119
蒋明,景耀先.胭脂鱼人工繁殖有关问题探析.重庆水产,2000,4:32-33
蒋文华,于道平.铜陵江段胭脂鱼资源现状与恢复的研究.特产研究,2003,25(3):31-33,46
金治萃,胡荫,白莲花,王陆一,李雪莲.桔皮注射液对免疫功能的影响.内蒙古医学杂志,1993,13(2):1-3
李爱华.我国鱼类病原菌耐药性、耐药质粒及几种药物抗菌作用的研究.[博士学位论文].武汉:中国科学院水生生物研究所,1998
李爱杰.水产动物营养与饲料学.北京:中国农业出版社,1994:8-104
李骏珉.胭脂鱼的人工繁殖试验报告.淡水渔业,1991,4:16-18
李年文.胭脂鱼的生物学特性及人工饲养技术(上).科学养鱼,1999a,7:12-13
李年文.胭脂鱼的生物学特性及人工饲养技术(下).科学养鱼,1999b,8:11-12
李荣,李俊,胡成穆,张磊,姜辉.橙皮苷对大鼠佐剂性关节炎的治疗作用及机制.中国药理学通报,2008,24(4):494-498
李荣,李俊,胡成穆,张磊,张茜.橙皮苷对免疫功能低下小鼠免疫调节作用的实验研究.中国药理学通报,2007,23(2):169-172
李树深,王蕊芳,刘光佐,王应祥,李崇云.中国胭脂鱼的核型研究.动物学研究,1983,4(2):182-183
李仪奎,姜名瑛.中药药理学.北京:中国中医药出版社,1992:192-194
李正秋,张奇亚.胭脂鱼弹状病毒包涵体在培养细胞中的形成.中国兽医学报,2001,21(1):38-41
刘恒,李光友.免疫多糖对养殖南美白对虾作用的研究.海洋与湖沼,1998,29(2):113-117
刘家国,胡元亮,张宝康,吕殿红,宋大鲁.淫羊藿-蜂胶佐剂对实验动物非特异免疫功能的影响.中国兽医学报,2001,21(5):489-493
刘家寿,余志堂.一龄胭脂鱼摄食与日粮的初步研究.水利渔业,1990,3:24-26
刘来亭,张慧茹,冯建新,马秋刚,计成.葎草醇提取物对草鱼非特异性免疫功能的影响.中国畜牧杂志,2008,44(9):29-31
刘树青,江晓路,牟海津,王慧谧,管华诗.免疫多糖对中国对虾血清溶菌酶、磷酸酶和过氧化物酶的作用.海洋与湖沼,1999,30(3):278-283
吕殿红.淫羊藿-蜂胶佐剂对雏鸡腹腔巨噬细胞吞噬功能的影响.中国兽医杂志,1999,25(3):46
马自佳.鱼病中药防治.北京:中国农业大学出版社,1998
米娜,吴敏毓,孙卫民.拆方研究补中益气汤中黄芪甘草及全方的免疫调节作用.皖南医学院学报,2002,21(1):13-15
钱国英.饵料中不同蛋白质、纤维素、脂肪水平对加州鲈鱼生长的影响.动物营养学报,2000,12(2):51-52
阙林林,陆建平,龚宏伟.长江胭脂鱼饵料转口培育试验.科学养鱼,2005,12:65-66
饶发祥.胭脂鱼的外部形态与生物学特性及其资源保护.江西水产科技,1993,2:31-34
阮国良,杨代勤,王金龙.几种中草药饲料添加剂对黄鳝免疫功能和生长性能的影响.饲料工业,2005,26(24):34-36
阮红梅,李正秋,张奇亚.胭脂鱼弹状病毒对鲤科鱼类感染性的测定.水生生物学报,2002,26(5):555-559
史春路.壳聚糖对黄颡鱼非特异性免疫机能和生长的影响.[硕士学位论文].武汉:华中农业大学图书馆,2008
宋学宏,蔡春芳,潘春法,赵林川.用生长和非特异性免疫力评定异育银鲫维生 素C需要量.水产学报,2002,(4):351-356
孙红梅,黄权,丛波,吕大成.饥饿对黄颡鱼免疫机能的影响.水利渔业,2006,26(3):80-81
孙红祥.一些中药及其挥发性成分抗霉菌活性研究.中国中药杂志,2001,26(2):99-102
孙峻岭,薛家宾,胡元亮,王德云,张宝康,刘家国.中药成分复方的佐剂作用及其与中药复方的功效比较.南京农业大学学报,2005,28(4):109-112
孙玉华.亚口鱼科鱼类分子系统学研究.水利渔业,2006,26(4):1-2,9
孙玉华,刘思阳,彭智,余来宁,吴清江.中国胭脂鱼种群的遗传分析.水生生物学报,2003,27(3):248-252
孙玉华,谢从新,刘思阳.亚口鱼科鱼类核DNA 18S-ITS1-5.8S序列比较分析.水生生物学报,2006,30(3):367-370
田杨星,杨军峰,汤明亮.长江中国胭脂鱼群体的遗传分化.武汉大学学报(理学版),2006,52(4):503-507
万全,赖年悦,刘映彬,沈保平,孙文贤,陈中华,王旭.配合饲料高密度培育一龄胭脂鱼种的研究.安徽农业科学,2006,18:4605-4606
万松良.胭脂鱼的生物学特性及养殖技术.农村养殖技术,2004,1:20-22
万松良,裴家田.胭脂鱼人工繁殖和鱼苗培育的初步研究.水利渔业,2002,22(2):1-2,22
王迪,张文风,张杰.黄芪配甘草对小鼠细胞免疫功能的影响.长春中医药大学学报,2006,22(3):42-43
王鸿泰.鱼类中草药药饵的配伍及制法.淡水渔业,1999,29(5):16-18
王吉桥,孙永新,张剑诚.金银花等复方草药对牙鲆生长、消化和免疫能力的影响.水产学报,2006,30(1):90-96
王雷,李光友,毛远兴.口服免疫性药物对中国对虾病害防治作用的研究.海洋与湖沼,1994,(25):481-491
王玲,董矜,汪德清,田亚平.黄芪提取物免疫调节作用的实验研究.军医进修学院学报,2005,26(4):293-295
王瑁,丘书院.体重和温度对花尾胡椒鲷幼鱼最大摄食量的研究.热带海洋学报, 2000,2(3):87-92
王天然,邢善田,周金黄.淫羊藿总黄酮促进免疫功能的实验.中成药研究,1987,(2):27-28
王文博.环境胁迫、中草药及基因转植对鱼体非特异性免疫功能的影响.[博士学位论文].武汉:中国科学院水生生物研究所,2005
王文博,方平,林小涛,夏磊,齐长宝,汪建国,孙军.甘草粗提物对鲫的免疫调节作用.水生生物学报,2007a,31(5):655-660
王文博,方平,林小涛,夏磊,齐长宝,汪建国,孙军.甘草粗提物对鲫的抗应激及免疫保护功效研究.淡水渔业,2007b,37(4):3-6
王文博,李爱华,汪建国,蔡桃珍,吴玉深.拥挤胁迫对草鱼非特异性免疫功能的影响.水产学报,2004,28(2):139-144
王文博,林小涛,方平,夏磊,齐长宝.腹注甘草粗提物对鲫免疫力的影响.中山大学学报(自然科学版),2007c,46(4):84-88
王宜艳,孙虎山,李光友.复合免疫药物对中国对虾血淋巴氧化酶和抗氧化酶活力的影响.海洋科学进展,2002,20(3):79-83
尾崎久雄.鱼类消化生理.上海:上海科学技术出版社,1985,365-370
吴凡.免疫刺激剂对水产动物免疫机能的影响.[硕士学位论文].武汉:华中农业大学图书馆,2005
项黎新,孟真,邵健忠.草鱼巨噬细胞活化的生理生化指标研究.浙江大学学报(理学版),2002,29(4):449-453
肖琳琳,张庆华.蜂胶佐剂对异育银鲫嗜水气单胞菌灭活疫苗的免疫增强试验.海洋渔业,2004,26(4):295-299
熊全沫,夏盛林.中国胭脂鱼同工酶的研究.动物学报,1985,31(1):20-27
薛治泉.胭脂鱼人工繁殖若干问题探讨.重庆水产,1998,3:40-42
杨静玉,于庆海,李爽,徐静华.淫羊藿总黄酮对免疫功能低下小鼠的免疫增强作用.沈阳药科大学学报,1998,15(2):94-97
杨龙圣,胡元亮,薛家宾,王芳,王德云,徐为中.中药成分复方对兔外周血淋巴细胞增殖、IFN-γ和IL-10的mRNA表达以及兔出血症疫苗免疫效果的影响.中国农业科学,2008,41(11):3809-3815
杨州,杨家新.暗纹东方纯幼鱼对蛋白质的最适需要量.水产学报,2003,27(5):452-455
袁春涛,孙方方,李冬梅,龚昳,吴光红,沈美芳,韩晓冬.饲喂两种中药配方对异育银鲫巨噬细胞免疫机能的影响.南京大学学报(自然科学),2007,43(2):152-156
张宝康,王德云,孔祥峰,胡元亮.蜂胶黄酮增强免疫作用的研究.扬州大学学报(农业与生命科学版),2006,27(3):20-22
张春光,赵亚辉,康景贵.我国胭脂鱼资源现状及其资源恢复途径的探讨.自然资源学报,2000,15(2):166-159
张明,蒋培红.中草药的免疫调节作用.动物医学进展,2006,27(11):41-44
张逸凡,于庆海.淫羊藿总黄酮的免疫调节作用.沈阳药科大学学报,1999,16(3):182-184
张友良,张飞明.不同食料对胭脂鱼鱼种生长的影响试验.水产科技情报,2006,33(2):59-60
中科院水生所.三峡工程生态环境监测的报告.武汉:中科院水生所,1996,1-30
周剑光,杨德国,吴国犀,王志玲,刘乐和,陈金生.胭脂鱼仔幼鱼发育及苗种培育技术.华中农业大学学报,1999,18(3):263-267
周显青,牛翠娟,孙儒永.黄芪对中华鳖免疫和抗酸应激能力的影响.水生生物学报.2003,27(1):110-112
Ainsworth A J.Fish granulocytes:morphology,distribution,and function.Ann Rev Fish Dis,1992,2:123-148
Ainsworth A J,Rice C D,Xue L.Immune responses of channel catfish,Ictalurus punctatus(Rafinesque),after oral or intraperitoneal vaccination with particulate or soluble Edwardsiella ictaluri antigen.J Fish Diseases,1995,18:397-409
Aksnes A,Hjertnes T,Opstvedt J.Effect of dietary protein level on growth and carcass composition in Atlantic halibut(Hippoglossus hippoglossus L.).Aquaculture,1996,145:225-233
Anderson D P.Immunostimulants,adjuvants,and vaccine careers in fish:application to aquaculture.Fish Diseases,1992(2):281-307
Anderson D P,Siwicki A K.Duration of Protection against Aeromonas salmonicida in brook trout immunostimulated with glucan or chitosan by injection or immersion.The Prog Fish Cult, 1996, 56(3): 258-261
Anderson R J, Kienholz E W, Flickinger S A. Protein requirements of smallmouth bass and largemouth bass. J Nutri, 1981, 11: 1085-1097
Arzel J, Metailler R, Kerleguer C, Le Delliou H, Guillaume J. The protein requirement of brown trout (Salmo trutta) fry. Aquaculture, 1995, 130: 67-68
Brecka B J, Kohler C C, Wahl D H. Effects of dietary protein concentration on growth,survival and body concentration of muskellunge Esox masquinongy and tiger muskellunge E. masquinongy X E. lucis fingerlings. J World Aquacult Soc, 1995,26:416-425
Chen C F, Chen X H, Riichi K. Adjuvant effect of glycyrrhizine in vaccines against bacterial septicemia in Mandarinfish, Siniperca chuatsi Basilewsky. 华中农业大学学报, 2000, 19(3): 256-260
Chen H Y, Tsai J C, Su M S. Optimal dietary protein level for the growth of juvenile grouper, Epinephelus malabaricus, fed semi-purified diets. Aquaculture, 1994,119:265-271
Christybapita D, Divyagnaneswari M, Michael R D. Oral administration of Eclipta alba leaf aqueous extract enhances the non-specific immune responses and disease resistance of Oreochromis mossambicus. Fish Shellfish Immunol, 2007,23: 840-852
Chung S, Secombes C J. Analysis of events occurring within teleost macrophages during the respiratory burst. Comp Biochem Physiol, 1988, 89(3): 539-544
Clark A E, Watanabe W O, Ahmed H, Olla B L, Wicklund R I. Growth, feed conversion and protein utilization of Florida red tilapia fed isocaloric diets with different protein levels in seawater pools. Aquaculture, 1990, 88: 75-85
Cuesta A, Rodriguez A, Esteban M A, Meseguer J. In vivo effects of propolis, a honeybee product, on gilthead seabream innate immune responses. Fish Shellfish Immunol, 2005,18:71-80
Dabrowski K. Protein requirement of grass carp (Ctenopharyngodon idella Val).Aquaculture, 1977, 12: 63-73
Elangovan A, Shim K F. Growth response of juvenile Barbodes altus fed isocaloric diets with variable protein levels. Aquaculture, 1997, 158: 321-329
Ellis A E. Lysozyme assays. In: Stolen J S, Fletcher T C, Anderson D P, Roberson B S,Muiswinkel, eds. Techniques in Fish Immunology I . USA: SOS Publications,1990, 101-103
Graham S, Jeffries A H, Secombes C J. A novel assay to detect macrophage bactericidal activity in fish: factors influencing the killing of Aeromonas salmonicida. J Fish Diseases, 1988, 11(4): 389-396
Grinde B. Lysozyme from rainbow trout, Salmo gairdneri Richardson, as an antibacterial agent against fish pathogens. J Fish Dis, 1989, 12: 95-104
Hofer R, Krewedl G, Koch F. An energy budget for an omnivorous cyprinid: Retilus rutilus (L.). Hydrobilogia, 1985, 122: 53-59
Islam M S, Tanaka M. Optimization of dietary protein requirement for pond-reared mahseer Tor putitora Hamilton (Cypriniformes: Cyprinidae). Aquacult Cult Res,2004,35: 1270-1276
Jauncey K. The effect of varying dietary protein level on the growth, food conversion,protein utilization and body composition of juvenile tilapia (Sarotherodon mossambicus). Aquaculture, 1982, 27: 43-54
Jezierska B. The effect of various type of food on the growth and chemical composition of the body of perch (Perca fluviatilis) in laboratory conditions. Pol Arch Hydrobiol, 1974,21:467-479
Jian J, Wu Z. Effects of traditional Chinese medicine on nonspecific immunity and disease resistance of large yellow croaker, Pseudosciaena crocea (Richardson).Aquaculture 2003, 218: 1-9
Jian J, Wu Z. Influence of traditional Chinese medicine on non specific immunity of Jian Carp (Cyprinus carpio var. Jian). Fish Shellfish Immunol, 2004, 16:185-191
Jose M V, Fernandez-Palacios H, Lidia R, Jauncey K, Manuel D L H, Marisol I. The effect of varying dietary protein level on the growth, feed efficiency, protein utilization and body composition of gilthead sea fry. Fish Sci, 1996, 62: 620-623
Khan M S, Ang K J, Ambak M A, Saad C R. Optimum dietary protein requirement of a Malaysian freshwater catfish, Mystus nemurus. Aquaculture, 1993, 112:227-235
Kim J D, Lall S P. Effects of dietary protein level on growth and utilization of protein and energy by juvenile haddock (Melanogrammus aeglefinus). Aquaculture,2001,195:311-319
Lazo J P, Davis D A, Arnold C R. The effects of dietary protein level on growth, feed efficiency and survival of juvenile Florida pompano (Trachinotus carolinus).Aquaculture, 1998, 169: 225-232
Li W, Asda Y, Yoshikawas T. Antimicrobial flavonoids from Glycyrrhiza glabrahairy root cultures. Planta Medica, 1998, 64: 746-747
Lochmann R T, Phillips H. Dietary protein requirement of juvenile golden shiners (Notemigonus crysoleucas) and goldfish (Carassiusaus ratus) in aquaria.Aquaculture, 1994, 128: 277-285
Millikin M R. Effects of dietary protein concentration on growth, feed efficiency and body composition of age-0 stripped bass. Trans Am Fish Soc, 1982, 111:373-378
National Research Council (NRC). Nutrient Requirements of Fish. Washington, DC:National Academy Press. 1993, 114
Nelson E M. Some notes on the Chinese sucker.Copeia, 1976, 3: 594-595
Ogunji J O, Wirth M. Effect of dietary protein content on growth, food conversion and body composition of tilapia Oreochromis niloticus fingerlings fed fish meal diet.Aqua trop, 2000, 15(4): 385-386
Olevera-Novoa M A, Gasca-leyva E, Martinez-Palacois C A. The dietary protein requirement of Cichlasoma synspilum Hubbs, 1935 (Pisces; Cichlidae) fry.Aquacult Cult Res, 1996,27: 167-173
Ortuno J, Esteban M A, Mesuguer J. Kinetics of hydrogen peroxide production during in vitro respiratory burst of seabream (Sparus aurata L.) head-kidney leucocytes,as measured by a flow cytometric method. Fish Shellfish Immunol, 2000, 10:725-729
Pickering A D, Potting T G, Christie P. Recovery of the brown trout, Salmo trutta L.,to chronic crowding stress. J Fish Biol, 1982, 20: 229-244
Pongmaneerat T, Watanabe T. Nutritional evaluation of soybean meal for rainbow trout and carp. Bull Jap Soc Sci Fish, 1993, 59: 157-163
Rao Y V, Das B K, Jyotyrmayee P, Chakrabarti R. Effect of Achyranthes aspera on the immunity and survival of Labeo rohita infected with Aeromonas hydrophila.Fish Shellfish Immunol, 2006, 20: 263-273
Robbins K R, Norton H R, Baker D H. Estimation of nutrient requirements from growth data. J Nutri, 1979, 109: 1710-1714
Samuel M, Lam T J, Sin Y M. Effect of laminaran [P-l,3-D-glucan] on the protective immunity of blue gourami, Trichogaster trichopterus against Aeromonas salmonicida. Fish Shellfish Immunol, 1996, 6(4): 443-454
Satia B P. Quantitative protein requirements of rainbow trout. Prog Fish Cult, 1974,36: 80-85
Secombes C J. Macrophage activation during experimental allergic orchitis in rainbow trout (Salmo gairdneri). Dev Com Immun, 1986, 10: 539-546
Secombes C J, Fletcher T C. The role of phagocytes in the protective mechanisms of fish. Annu Rev Fish Dis, 1992, 2: 53-71
Sforcin J M. Propolis and the immune system: a review. J Ethnopharmacol, 2007, 113:1-14
Shearer K D. Factors affecting the proximate composition of cultured fishes with emphasis on salmonids. Aquaculture, 1994, 119: 63-88
Shiau S Y, Huang S L. Optimal dietary protein for hybrid telapia (Oreochromis niloticus). Aquaculture, 1989, 70: 63-73
Shiau S Y, Lan C W. Optimum dietary protein level and protein to energy ratio for growth of grouper (Epinephelus malabarricus). Aquaculture, 1996, 70: 63-73
Shim K J, Jung K H, Chung M K. Development of an in vitro environmental monitoring system by using immune cells. J Health Sci, 2002, 48(2): 130-133
Siddiqui A Q, Howlader M S, Adam A A. Effects dietary protein levels on growth,feed conversion and protein utilization in fry and young Nile tiapia, Oreochromis nilotica. Aquaculture, 1988, 70: 63-73
Song Zhao-bin, Fu Zi-dong, Li Jing, Yue Bi-song. Validation of daily otolith increments in larval and juvenile Chinese sucker, Myxocyprinus asiaticus.Environmental Biology of Fishes, 2008, 82(2): 165-171
Song Zhao-bin, He Chun-lin, Fu Zi-dong, Shen Dan-zhou. Otolith thermal marking in larval Chinese sucker, Myxocyprinus asiaticus.Environmental Biology of Fishes,2008,82(1): 1-7
Sovenyi J, Szakolczai J. Studies on the toxic and immunosuppressive effects of cadmium on the common carp. Acta vet Hung, 1993,41(3-4): 415-426
Tibbetts S M, Lall S P, Anderson D M. Dietary protein requirement of juvenile American eel (Anguilla rostrata) fed practical diets. Aquaculture, 2000, 186:145-155
Tsoi S, Sin-che Lee, Wei-chung Chao. Duplicate gene expression anbd diploidization in an asian tetraploid catostomid, Myxocyprinus asiaticus (Cypriniformes,Catostomidae). Comparative biochemistry and physiology. B. Comparative biochemistry, 1989, 93(1): 27-32
Van der Meer M B, Machiels M A M, Verdegem M C J. The effect of dietary protein level on growth, protein utilization and body composition of Cichlasoma synspilum (Cuvier). Aquacult Cult Res, 1995, 26: 901-909
Wan Qiu-hong, Fang Sheng-guo. Reduced genetic diversity in a breeding population of the Chinese sucker (Myxocyrinus asiaticus Bleeker) as revealed by DNA fingerprinting. J Fresh Ecol, 2002, 17(2): 269-274
Ward L R, Carter C G, Crear B J, Smith D M. Optimal dietary protein level for juvenile southern rock lobster, Jasus edwardsii, at two lipid levels. Aquaculture,2003,217:483-500
Yano T. Assays of hemolytic complement activity. In: Stolen J S, Fletcher T C,Anderson D P, Roberson B S, Muiswinkel, eds. Techniques in Fish Immunology II, USA: SOS Publications, 1992, 131-141
Yin G, Ardo L, Thompson K D, Adams A, Jeney Z, Jeney G. Chinese herbs (Astragalus radix and ganoderma lucidum) enhance immune response of carp, cyprinus carpio,and protection against aeromonas hydrophila.Fish Shellfish Immunol,2009,26(1):140-145
Yin G,Jeney G,Racz T,Xu P,Jun X,Jeney Z.Effect of two Chinese herbs (Astragalus radix and Scutellaria radix) on non specific immune response of tilapia,Oreochromis niloticus.Aquaculture,2006,253:39-47
Yin G J,Geert W,Li Y M,Johan S,Johan V,Xu P,Zhou H Q.Effect of Astragalus radixon proliferation and nitric oxide production of head kidney macrophages in Cyprinus carpio:an in vitro study.水产学报,2004,28(6):628-632
Zhang Qi-Ya,Li Zheng-Qiu,Gui Jian-Fang.Isolation of a lethal rhabdovirus from the cultured Chinese sucker Myxocyprinus asiaticus.Dis Aquat Organ,2000,42(1):1-9
蔡春芳,宋学宏,潘兴法,成文靖,浦佩,朱素琴.几种抗病促生长剂对银鲫生长和免疫力的影响.水利渔业,2002,22(2):20-22
曹丽萍,丁炜东,张柳,Galina J,徐跑,殷国俊.香菇多糖和黄芪多糖对鲤免疫细胞的活性和IL-1β体外诱生表达的影响.水产学报,2008a,32(4):628-635
曹丽萍,丁炜东,张柳,Galina J,殷国俊.2种多糖对鲤鱼离体培养免疫细胞活性的影响.安徽农业大学学报,2008b,35(2):219-223
曹振杰,曲世科,丛日祥,马俊岭,王志忠,刘汉华,张家国,孙鹤田,轩子群.免疫多糖对草鱼免疫功能的影响.齐鲁渔业,1999,16(3):43-44
陈昌福,罗宇良,蔡冰.饲养水温对草鱼溶菌酶活性的影响.中国水产科学,1996,3(3):24-30
陈超然,陈晓辉,陈昌福.口服甘草素对中华鳖稚鳖抗嗜水气单胞菌感染的作用.华中农业大学学报,2000,19(6):577-580
陈家长,简纪常.水体中NH3-N对建鲤非特异性免疫功能的影响.湛江海洋大学学报,2000,20(3):13-16
陈金生,刘家寿.不同饲料养殖一龄胭脂鱼种生长试验.水利渔业,1992,4:9-11,14
陈忻,黄耀威,赖兴华,杨桂芳.鱼类饲料甘草添加剂的研究.佛山科学技术学院学报(自然科学版),1999,17(3):36-38
陈孝煊,吴志新,殷居易,李莉.大黄、穿心莲、板蓝根和金银花对异育银鲫免疫机能的影响.中国水产科学,2003,10(1):36-39
陈孝煊,吴志新,张厚梅.大黄与黄连对二种淡水虾血细胞吞噬活性的影响.水生生物学报,2002,26(2):201-204
陈兆.胭脂鱼的生物学特性及饲养技术.渔业致富指南,2005,22:40-43
陈振昆,丁光.陈皮对草鱼诱食作用的研究.云南农业大学学报,1996,11(1):35-37
邓中群,余志堂,赵燕.三峡水利枢纽对长江白鲟和胭脂鱼影响的评价及资源保护的研究.见:长江三峡工程对生志与环境影响及其对策研究论文集.北京:科学出版社,1987,42-52
丁光,陈振昆,李梅,李国治.饲料添加剂陈皮对草鱼淋巴细胞转化率影响的研究.云南农业大学学报,2003,18(4):401-403
丁光,陈振昆,李文贵,杨贵树,许春艳.陈皮对草鱼细胞免疫影响的研究.云南农业大学学报,2004,19(6):727-730
丁光,陈振昆,徐宝明.在饲料中添加柑橘或甜橙外果皮对鲤鱼诱食作用的比较研究.水产科学,1999,18(1):16-19
杜爱芳,叶均安,于涟.复方大蒜油添加剂对中国对虾免疫机能的增强作用.浙江农业大学学报,1997,23(3):317-320
付自东,李静,岳碧松,宋昭彬.用荧光物质浸泡标记胭脂鱼仔、稚鱼耳石.动物学杂志,2005,40(4):60-65
龚宏伟,蔡春芳,阙林林.长江胭脂鱼开口饵料的研究.水产科学,2005,24(11):7-9
郭宝林,余竞光,肖培根.淫羊霍化学成分的研究.中国中药杂志,1996,21(5):294-296
郭文婷.两种免疫增强剂对牙鲆和凡纳滨对虾免疫因子影响的研究.f硕士学位论文].青岛:中国海洋大学,2005
郭永军,刑克智,陈成勋,杨羽兴,朱国霞.几种中草药对鲤鱼诱食效果的研究.天津农学院学报,2005,12(3):1-5
郝炳华,陶明,何立端,薛晚利,李选社,孟晓军,张京田.淫羊藿总黄酮对荷瘤小鼠细胞免疫及红细胞免疫功能的影响.西安医科大学学报,1999,20(3):323-325
胡兵,刘军,侯永清,李惠,陈爱敬,辜玲芳.黄芪多糖对异育银鲫非特异性免疫力的影响.水利渔业,2008,28(3):108-111
湖南省水产科学研究所.湖南鱼类志.长沙:湖南科学技术出版社,1980,24-25.
胡隐昌.胭脂鱼的主要生物学.珠江水产,2001,2:33-35
胡元亮,刘家国,宋大鲁,许志刚.淫蜂佐剂影响动物免疫功能的实验研究.畜 牧兽医学报,2000,31(2):155-160
胡元亮,吕殿红,刘家国,宋大鲁.淫羊藿.蜂胶佐剂对雏鸡和小白鼠腹腔巨噬细胞活性的影响.中国兽医科技,1999,29(5):8-10
黄克安,朱木洛,吴杏清.莨菪类药物防治草鱼出血病试验.鱼病简讯,1985,(1):20-23
黄小丽,邓永强.中草药免疫增强剂在水产上的开发应用.内陆水产,2004,8:24-26
吉红,李海梅,周继术,刘兰锋.4种植物粗提物对金鱼诱食效果的初步研究.水产科学,2008,27(2):67-70
简纪常,吴灶和.中草药对建鲤非特异性免疫功能的影响.大连水产学院学报,2002,17(2):114-119
蒋明,景耀先.胭脂鱼人工繁殖有关问题探析.重庆水产,2000,4:32-33
蒋文华,于道平.铜陵江段胭脂鱼资源现状与恢复的研究.特产研究,2003,25(3):31-33,46
金治萃,胡荫,白莲花,王陆一,李雪莲.桔皮注射液对免疫功能的影响.内蒙古医学杂志,1993,13(2):1-3
李爱华.我国鱼类病原菌耐药性、耐药质粒及几种药物抗菌作用的研究.[博士学位论文].武汉:中国科学院水生生物研究所,1998
李爱杰.水产动物营养与饲料学.北京:中国农业出版社,1994:8-104
李骏珉.胭脂鱼的人工繁殖试验报告.淡水渔业,1991,4:16-18
李年文.胭脂鱼的生物学特性及人工饲养技术(上).科学养鱼,1999a,7:12-13
李年文.胭脂鱼的生物学特性及人工饲养技术(下).科学养鱼,1999b,8:11-12
李荣,李俊,胡成穆,张磊,姜辉.橙皮苷对大鼠佐剂性关节炎的治疗作用及机制.中国药理学通报,2008,24(4):494-498
李荣,李俊,胡成穆,张磊,张茜.橙皮苷对免疫功能低下小鼠免疫调节作用的实验研究.中国药理学通报,2007,23(2):169-172
李树深,王蕊芳,刘光佐,王应祥,李崇云.中国胭脂鱼的核型研究.动物学研究,1983,4(2):182-183
李仪奎,姜名瑛.中药药理学.北京:中国中医药出版社,1992:192-194
李正秋,张奇亚.胭脂鱼弹状病毒包涵体在培养细胞中的形成.中国兽医学报,2001,21(1):38-41
刘恒,李光友.免疫多糖对养殖南美白对虾作用的研究.海洋与湖沼,1998,29(2):113-117
刘家国,胡元亮,张宝康,吕殿红,宋大鲁.淫羊藿-蜂胶佐剂对实验动物非特异免疫功能的影响.中国兽医学报,2001,21(5):489-493
刘家寿,余志堂.一龄胭脂鱼摄食与日粮的初步研究.水利渔业,1990,3:24-26
刘来亭,张慧茹,冯建新,马秋刚,计成.葎草醇提取物对草鱼非特异性免疫功能的影响.中国畜牧杂志,2008,44(9):29-31
刘树青,江晓路,牟海津,王慧谧,管华诗.免疫多糖对中国对虾血清溶菌酶、磷酸酶和过氧化物酶的作用.海洋与湖沼,1999,30(3):278-283
吕殿红.淫羊藿-蜂胶佐剂对雏鸡腹腔巨噬细胞吞噬功能的影响.中国兽医杂志,1999,25(3):46
马自佳.鱼病中药防治.北京:中国农业大学出版社,1998
米娜,吴敏毓,孙卫民.拆方研究补中益气汤中黄芪甘草及全方的免疫调节作用.皖南医学院学报,2002,21(1):13-15
钱国英.饵料中不同蛋白质、纤维素、脂肪水平对加州鲈鱼生长的影响.动物营养学报,2000,12(2):51-52
阙林林,陆建平,龚宏伟.长江胭脂鱼饵料转口培育试验.科学养鱼,2005,12:65-66
饶发祥.胭脂鱼的外部形态与生物学特性及其资源保护.江西水产科技,1993,2:31-34
阮国良,杨代勤,王金龙.几种中草药饲料添加剂对黄鳝免疫功能和生长性能的影响.饲料工业,2005,26(24):34-36
阮红梅,李正秋,张奇亚.胭脂鱼弹状病毒对鲤科鱼类感染性的测定.水生生物学报,2002,26(5):555-559
史春路.壳聚糖对黄颡鱼非特异性免疫机能和生长的影响.[硕士学位论文].武汉:华中农业大学图书馆,2008
宋学宏,蔡春芳,潘春法,赵林川.用生长和非特异性免疫力评定异育银鲫维生 素C需要量.水产学报,2002,(4):351-356
孙红梅,黄权,丛波,吕大成.饥饿对黄颡鱼免疫机能的影响.水利渔业,2006,26(3):80-81
孙红祥.一些中药及其挥发性成分抗霉菌活性研究.中国中药杂志,2001,26(2):99-102
孙峻岭,薛家宾,胡元亮,王德云,张宝康,刘家国.中药成分复方的佐剂作用及其与中药复方的功效比较.南京农业大学学报,2005,28(4):109-112
孙玉华.亚口鱼科鱼类分子系统学研究.水利渔业,2006,26(4):1-2,9
孙玉华,刘思阳,彭智,余来宁,吴清江.中国胭脂鱼种群的遗传分析.水生生物学报,2003,27(3):248-252
孙玉华,谢从新,刘思阳.亚口鱼科鱼类核DNA 18S-ITS1-5.8S序列比较分析.水生生物学报,2006,30(3):367-370
田杨星,杨军峰,汤明亮.长江中国胭脂鱼群体的遗传分化.武汉大学学报(理学版),2006,52(4):503-507
万全,赖年悦,刘映彬,沈保平,孙文贤,陈中华,王旭.配合饲料高密度培育一龄胭脂鱼种的研究.安徽农业科学,2006,18:4605-4606
万松良.胭脂鱼的生物学特性及养殖技术.农村养殖技术,2004,1:20-22
万松良,裴家田.胭脂鱼人工繁殖和鱼苗培育的初步研究.水利渔业,2002,22(2):1-2,22
王迪,张文风,张杰.黄芪配甘草对小鼠细胞免疫功能的影响.长春中医药大学学报,2006,22(3):42-43
王鸿泰.鱼类中草药药饵的配伍及制法.淡水渔业,1999,29(5):16-18
王吉桥,孙永新,张剑诚.金银花等复方草药对牙鲆生长、消化和免疫能力的影响.水产学报,2006,30(1):90-96
王雷,李光友,毛远兴.口服免疫性药物对中国对虾病害防治作用的研究.海洋与湖沼,1994,(25):481-491
王玲,董矜,汪德清,田亚平.黄芪提取物免疫调节作用的实验研究.军医进修学院学报,2005,26(4):293-295
王瑁,丘书院.体重和温度对花尾胡椒鲷幼鱼最大摄食量的研究.热带海洋学报, 2000,2(3):87-92
王天然,邢善田,周金黄.淫羊藿总黄酮促进免疫功能的实验.中成药研究,1987,(2):27-28
王文博.环境胁迫、中草药及基因转植对鱼体非特异性免疫功能的影响.[博士学位论文].武汉:中国科学院水生生物研究所,2005
王文博,方平,林小涛,夏磊,齐长宝,汪建国,孙军.甘草粗提物对鲫的免疫调节作用.水生生物学报,2007a,31(5):655-660
王文博,方平,林小涛,夏磊,齐长宝,汪建国,孙军.甘草粗提物对鲫的抗应激及免疫保护功效研究.淡水渔业,2007b,37(4):3-6
王文博,李爱华,汪建国,蔡桃珍,吴玉深.拥挤胁迫对草鱼非特异性免疫功能的影响.水产学报,2004,28(2):139-144
王文博,林小涛,方平,夏磊,齐长宝.腹注甘草粗提物对鲫免疫力的影响.中山大学学报(自然科学版),2007c,46(4):84-88
王宜艳,孙虎山,李光友.复合免疫药物对中国对虾血淋巴氧化酶和抗氧化酶活力的影响.海洋科学进展,2002,20(3):79-83
尾崎久雄.鱼类消化生理.上海:上海科学技术出版社,1985,365-370
吴凡.免疫刺激剂对水产动物免疫机能的影响.[硕士学位论文].武汉:华中农业大学图书馆,2005
项黎新,孟真,邵健忠.草鱼巨噬细胞活化的生理生化指标研究.浙江大学学报(理学版),2002,29(4):449-453
肖琳琳,张庆华.蜂胶佐剂对异育银鲫嗜水气单胞菌灭活疫苗的免疫增强试验.海洋渔业,2004,26(4):295-299
熊全沫,夏盛林.中国胭脂鱼同工酶的研究.动物学报,1985,31(1):20-27
薛治泉.胭脂鱼人工繁殖若干问题探讨.重庆水产,1998,3:40-42
杨静玉,于庆海,李爽,徐静华.淫羊藿总黄酮对免疫功能低下小鼠的免疫增强作用.沈阳药科大学学报,1998,15(2):94-97
杨龙圣,胡元亮,薛家宾,王芳,王德云,徐为中.中药成分复方对兔外周血淋巴细胞增殖、IFN-γ和IL-10的mRNA表达以及兔出血症疫苗免疫效果的影响.中国农业科学,2008,41(11):3809-3815
杨州,杨家新.暗纹东方纯幼鱼对蛋白质的最适需要量.水产学报,2003,27(5):452-455
袁春涛,孙方方,李冬梅,龚昳,吴光红,沈美芳,韩晓冬.饲喂两种中药配方对异育银鲫巨噬细胞免疫机能的影响.南京大学学报(自然科学),2007,43(2):152-156
张宝康,王德云,孔祥峰,胡元亮.蜂胶黄酮增强免疫作用的研究.扬州大学学报(农业与生命科学版),2006,27(3):20-22
张春光,赵亚辉,康景贵.我国胭脂鱼资源现状及其资源恢复途径的探讨.自然资源学报,2000,15(2):166-159
张明,蒋培红.中草药的免疫调节作用.动物医学进展,2006,27(11):41-44
张逸凡,于庆海.淫羊藿总黄酮的免疫调节作用.沈阳药科大学学报,1999,16(3):182-184
张友良,张飞明.不同食料对胭脂鱼鱼种生长的影响试验.水产科技情报,2006,33(2):59-60
中科院水生所.三峡工程生态环境监测的报告.武汉:中科院水生所,1996,1-30
周剑光,杨德国,吴国犀,王志玲,刘乐和,陈金生.胭脂鱼仔幼鱼发育及苗种培育技术.华中农业大学学报,1999,18(3):263-267
周显青,牛翠娟,孙儒永.黄芪对中华鳖免疫和抗酸应激能力的影响.水生生物学报.2003,27(1):110-112
Ainsworth A J.Fish granulocytes:morphology,distribution,and function.Ann Rev Fish Dis,1992,2:123-148
Ainsworth A J,Rice C D,Xue L.Immune responses of channel catfish,Ictalurus punctatus(Rafinesque),after oral or intraperitoneal vaccination with particulate or soluble Edwardsiella ictaluri antigen.J Fish Diseases,1995,18:397-409
Aksnes A,Hjertnes T,Opstvedt J.Effect of dietary protein level on growth and carcass composition in Atlantic halibut(Hippoglossus hippoglossus L.).Aquaculture,1996,145:225-233
Anderson D P.Immunostimulants,adjuvants,and vaccine careers in fish:application to aquaculture.Fish Diseases,1992(2):281-307
Anderson D P,Siwicki A K.Duration of Protection against Aeromonas salmonicida in brook trout immunostimulated with glucan or chitosan by injection or immersion.The Prog Fish Cult, 1996, 56(3): 258-261
Anderson R J, Kienholz E W, Flickinger S A. Protein requirements of smallmouth bass and largemouth bass. J Nutri, 1981, 11: 1085-1097
Arzel J, Metailler R, Kerleguer C, Le Delliou H, Guillaume J. The protein requirement of brown trout (Salmo trutta) fry. Aquaculture, 1995, 130: 67-68
Brecka B J, Kohler C C, Wahl D H. Effects of dietary protein concentration on growth,survival and body concentration of muskellunge Esox masquinongy and tiger muskellunge E. masquinongy X E. lucis fingerlings. J World Aquacult Soc, 1995,26:416-425
Chen C F, Chen X H, Riichi K. Adjuvant effect of glycyrrhizine in vaccines against bacterial septicemia in Mandarinfish, Siniperca chuatsi Basilewsky. 华中农业大学学报, 2000, 19(3): 256-260
Chen H Y, Tsai J C, Su M S. Optimal dietary protein level for the growth of juvenile grouper, Epinephelus malabaricus, fed semi-purified diets. Aquaculture, 1994,119:265-271
Christybapita D, Divyagnaneswari M, Michael R D. Oral administration of Eclipta alba leaf aqueous extract enhances the non-specific immune responses and disease resistance of Oreochromis mossambicus. Fish Shellfish Immunol, 2007,23: 840-852
Chung S, Secombes C J. Analysis of events occurring within teleost macrophages during the respiratory burst. Comp Biochem Physiol, 1988, 89(3): 539-544
Clark A E, Watanabe W O, Ahmed H, Olla B L, Wicklund R I. Growth, feed conversion and protein utilization of Florida red tilapia fed isocaloric diets with different protein levels in seawater pools. Aquaculture, 1990, 88: 75-85
Cuesta A, Rodriguez A, Esteban M A, Meseguer J. In vivo effects of propolis, a honeybee product, on gilthead seabream innate immune responses. Fish Shellfish Immunol, 2005,18:71-80
Dabrowski K. Protein requirement of grass carp (Ctenopharyngodon idella Val).Aquaculture, 1977, 12: 63-73
Elangovan A, Shim K F. Growth response of juvenile Barbodes altus fed isocaloric diets with variable protein levels. Aquaculture, 1997, 158: 321-329
Ellis A E. Lysozyme assays. In: Stolen J S, Fletcher T C, Anderson D P, Roberson B S,Muiswinkel, eds. Techniques in Fish Immunology I . USA: SOS Publications,1990, 101-103
Graham S, Jeffries A H, Secombes C J. A novel assay to detect macrophage bactericidal activity in fish: factors influencing the killing of Aeromonas salmonicida. J Fish Diseases, 1988, 11(4): 389-396
Grinde B. Lysozyme from rainbow trout, Salmo gairdneri Richardson, as an antibacterial agent against fish pathogens. J Fish Dis, 1989, 12: 95-104
Hofer R, Krewedl G, Koch F. An energy budget for an omnivorous cyprinid: Retilus rutilus (L.). Hydrobilogia, 1985, 122: 53-59
Islam M S, Tanaka M. Optimization of dietary protein requirement for pond-reared mahseer Tor putitora Hamilton (Cypriniformes: Cyprinidae). Aquacult Cult Res,2004,35: 1270-1276
Jauncey K. The effect of varying dietary protein level on the growth, food conversion,protein utilization and body composition of juvenile tilapia (Sarotherodon mossambicus). Aquaculture, 1982, 27: 43-54
Jezierska B. The effect of various type of food on the growth and chemical composition of the body of perch (Perca fluviatilis) in laboratory conditions. Pol Arch Hydrobiol, 1974,21:467-479
Jian J, Wu Z. Effects of traditional Chinese medicine on nonspecific immunity and disease resistance of large yellow croaker, Pseudosciaena crocea (Richardson).Aquaculture 2003, 218: 1-9
Jian J, Wu Z. Influence of traditional Chinese medicine on non specific immunity of Jian Carp (Cyprinus carpio var. Jian). Fish Shellfish Immunol, 2004, 16:185-191
Jose M V, Fernandez-Palacios H, Lidia R, Jauncey K, Manuel D L H, Marisol I. The effect of varying dietary protein level on the growth, feed efficiency, protein utilization and body composition of gilthead sea fry. Fish Sci, 1996, 62: 620-623
Khan M S, Ang K J, Ambak M A, Saad C R. Optimum dietary protein requirement of a Malaysian freshwater catfish, Mystus nemurus. Aquaculture, 1993, 112:227-235
Kim J D, Lall S P. Effects of dietary protein level on growth and utilization of protein and energy by juvenile haddock (Melanogrammus aeglefinus). Aquaculture,2001,195:311-319
Lazo J P, Davis D A, Arnold C R. The effects of dietary protein level on growth, feed efficiency and survival of juvenile Florida pompano (Trachinotus carolinus).Aquaculture, 1998, 169: 225-232
Li W, Asda Y, Yoshikawas T. Antimicrobial flavonoids from Glycyrrhiza glabrahairy root cultures. Planta Medica, 1998, 64: 746-747
Lochmann R T, Phillips H. Dietary protein requirement of juvenile golden shiners (Notemigonus crysoleucas) and goldfish (Carassiusaus ratus) in aquaria.Aquaculture, 1994, 128: 277-285
Millikin M R. Effects of dietary protein concentration on growth, feed efficiency and body composition of age-0 stripped bass. Trans Am Fish Soc, 1982, 111:373-378
National Research Council (NRC). Nutrient Requirements of Fish. Washington, DC:National Academy Press. 1993, 114
Nelson E M. Some notes on the Chinese sucker.Copeia, 1976, 3: 594-595
Ogunji J O, Wirth M. Effect of dietary protein content on growth, food conversion and body composition of tilapia Oreochromis niloticus fingerlings fed fish meal diet.Aqua trop, 2000, 15(4): 385-386
Olevera-Novoa M A, Gasca-leyva E, Martinez-Palacois C A. The dietary protein requirement of Cichlasoma synspilum Hubbs, 1935 (Pisces; Cichlidae) fry.Aquacult Cult Res, 1996,27: 167-173
Ortuno J, Esteban M A, Mesuguer J. Kinetics of hydrogen peroxide production during in vitro respiratory burst of seabream (Sparus aurata L.) head-kidney leucocytes,as measured by a flow cytometric method. Fish Shellfish Immunol, 2000, 10:725-729
Pickering A D, Potting T G, Christie P. Recovery of the brown trout, Salmo trutta L.,to chronic crowding stress. J Fish Biol, 1982, 20: 229-244
Pongmaneerat T, Watanabe T. Nutritional evaluation of soybean meal for rainbow trout and carp. Bull Jap Soc Sci Fish, 1993, 59: 157-163
Rao Y V, Das B K, Jyotyrmayee P, Chakrabarti R. Effect of Achyranthes aspera on the immunity and survival of Labeo rohita infected with Aeromonas hydrophila.Fish Shellfish Immunol, 2006, 20: 263-273
Robbins K R, Norton H R, Baker D H. Estimation of nutrient requirements from growth data. J Nutri, 1979, 109: 1710-1714
Samuel M, Lam T J, Sin Y M. Effect of laminaran [P-l,3-D-glucan] on the protective immunity of blue gourami, Trichogaster trichopterus against Aeromonas salmonicida. Fish Shellfish Immunol, 1996, 6(4): 443-454
Satia B P. Quantitative protein requirements of rainbow trout. Prog Fish Cult, 1974,36: 80-85
Secombes C J. Macrophage activation during experimental allergic orchitis in rainbow trout (Salmo gairdneri). Dev Com Immun, 1986, 10: 539-546
Secombes C J, Fletcher T C. The role of phagocytes in the protective mechanisms of fish. Annu Rev Fish Dis, 1992, 2: 53-71
Sforcin J M. Propolis and the immune system: a review. J Ethnopharmacol, 2007, 113:1-14
Shearer K D. Factors affecting the proximate composition of cultured fishes with emphasis on salmonids. Aquaculture, 1994, 119: 63-88
Shiau S Y, Huang S L. Optimal dietary protein for hybrid telapia (Oreochromis niloticus). Aquaculture, 1989, 70: 63-73
Shiau S Y, Lan C W. Optimum dietary protein level and protein to energy ratio for growth of grouper (Epinephelus malabarricus). Aquaculture, 1996, 70: 63-73
Shim K J, Jung K H, Chung M K. Development of an in vitro environmental monitoring system by using immune cells. J Health Sci, 2002, 48(2): 130-133
Siddiqui A Q, Howlader M S, Adam A A. Effects dietary protein levels on growth,feed conversion and protein utilization in fry and young Nile tiapia, Oreochromis nilotica. Aquaculture, 1988, 70: 63-73
Song Zhao-bin, Fu Zi-dong, Li Jing, Yue Bi-song. Validation of daily otolith increments in larval and juvenile Chinese sucker, Myxocyprinus asiaticus.Environmental Biology of Fishes, 2008, 82(2): 165-171
Song Zhao-bin, He Chun-lin, Fu Zi-dong, Shen Dan-zhou. Otolith thermal marking in larval Chinese sucker, Myxocyprinus asiaticus.Environmental Biology of Fishes,2008,82(1): 1-7
Sovenyi J, Szakolczai J. Studies on the toxic and immunosuppressive effects of cadmium on the common carp. Acta vet Hung, 1993,41(3-4): 415-426
Tibbetts S M, Lall S P, Anderson D M. Dietary protein requirement of juvenile American eel (Anguilla rostrata) fed practical diets. Aquaculture, 2000, 186:145-155
Tsoi S, Sin-che Lee, Wei-chung Chao. Duplicate gene expression anbd diploidization in an asian tetraploid catostomid, Myxocyprinus asiaticus (Cypriniformes,Catostomidae). Comparative biochemistry and physiology. B. Comparative biochemistry, 1989, 93(1): 27-32
Van der Meer M B, Machiels M A M, Verdegem M C J. The effect of dietary protein level on growth, protein utilization and body composition of Cichlasoma synspilum (Cuvier). Aquacult Cult Res, 1995, 26: 901-909
Wan Qiu-hong, Fang Sheng-guo. Reduced genetic diversity in a breeding population of the Chinese sucker (Myxocyrinus asiaticus Bleeker) as revealed by DNA fingerprinting. J Fresh Ecol, 2002, 17(2): 269-274
Ward L R, Carter C G, Crear B J, Smith D M. Optimal dietary protein level for juvenile southern rock lobster, Jasus edwardsii, at two lipid levels. Aquaculture,2003,217:483-500
Yano T. Assays of hemolytic complement activity. In: Stolen J S, Fletcher T C,Anderson D P, Roberson B S, Muiswinkel, eds. Techniques in Fish Immunology II, USA: SOS Publications, 1992, 131-141
Yin G, Ardo L, Thompson K D, Adams A, Jeney Z, Jeney G. Chinese herbs (Astragalus radix and ganoderma lucidum) enhance immune response of carp, cyprinus carpio,and protection against aeromonas hydrophila.Fish Shellfish Immunol,2009,26(1):140-145
Yin G,Jeney G,Racz T,Xu P,Jun X,Jeney Z.Effect of two Chinese herbs (Astragalus radix and Scutellaria radix) on non specific immune response of tilapia,Oreochromis niloticus.Aquaculture,2006,253:39-47
Yin G J,Geert W,Li Y M,Johan S,Johan V,Xu P,Zhou H Q.Effect of Astragalus radixon proliferation and nitric oxide production of head kidney macrophages in Cyprinus carpio:an in vitro study.水产学报,2004,28(6):628-632
Zhang Qi-Ya,Li Zheng-Qiu,Gui Jian-Fang.Isolation of a lethal rhabdovirus from the cultured Chinese sucker Myxocyprinus asiaticus.Dis Aquat Organ,2000,42(1):1-9