苏云金芽孢杆菌对黄河鲤血液免疫效果研究

研究了苏云金芽孢杆菌对黄河鲤血液免疫应答的影响.将苏云金芽孢杆菌按1.0×10~(11)、3.0×10~(11)、5.0×10~(11) cfu/kg 3个浓度添加在鱼用全价颗粒饲料中,投喂经嗜水气单胞菌疫苗免疫组(A_0～A_3)和未免疫组(B_0～B_3)黄河鲤,分别于0、15、30、40 d时检测各组黄河鲤白细胞吞噬活性、血清溶菌酶活性以及血清凝集抗体效价,并进行攻毒试验.结果表明:添加苏云金芽孢杆菌可使黄河鲤血液中白细胞吞噬活性和血清溶菌酶活性显著提高.添加组与对照组差异显著(P<0.05),添加浓度越高,血液白细胞活性和溶菌酶活性越强.免疫组与未免疫组白细胞吞噬活性差异不显著(P>0.05),而溶菌酶活性差异显著(P<0.05).在停止投喂后10 d,白细胞活性、血清溶菌酶活性逐渐下降,但差异不显著(P>0.05).添加苏云金芽孢杆菌还可提高黄河鲤的凝集抗体效价及攻毒后的存活率,其中,当菌株添加浓度为5.0×10~(11) cfu/kg时,受免疫的黄河鲤免疫保护率最高.即投喂苏云金芽孢杆菌对黄河鲤血液免疫应答有明显促进作用.     

鲤IGF-Ⅰ基因2个高度多态微卫星位点的鉴定及其与生长相关性状的关联分析

利用PCR结合测序方法,在鲤(Cyprinus carpio)类胰岛素生长因子-Ⅰ(insulin-like growth factor-Ⅰ,IGF-Ⅰ)基因内含子1和内含子2中各鉴定1个微卫星多态性位点,分别命名为intron1189和intron2310,这两个位点均以4碱基为重复单元.以黑龙江鲤(Cyprinus cario haematopterus,YL)(n=263)、德国镜鲤选育系(Cyprinus carpio L.mirror,JL)(n=229),及荷包红鲤抗寒品系(Cyprinus carpio var.wuyuanensis,HL)(n=255)为研究对象,评估了这两个位点不同基因型(频率＞3％)与鲤4个生长时段(135 d、325 d、335 d和445 d)生长表型的关联.结果显示,2个位点在3个鲤群体中均表现为高度多态(PIC＞0.5).intron1189位点多态性主要对YL的135 d、325 d体长和325 d、385 d体质量有显著影响(P＜0.05),而intron2 310位点多态性对JL各检测时段的体长和体质量均有显著影响(P＜0.05).对不同基因型个体的体长和体质量性状进行多重比较,结果显示,在intron 1189位点,185/229基因型YL的135 d、325 d体长和325 d、385 d体质量最低,而205/221基因型YL的135 d、325 d体长和325 d体质量最高.在mtron2310位点,290/350基因型JL在各检测时段的体长及135 d、325 d、385 d体质量最低,而318/350基因型JL的135 d、325 d、385 d体长和体质量均为最高.上述结果表明,鲤IGF-Ⅰ基因内含子中的这两个高度多态微卫星位点潜在影响鲤的生长性能.     

鲤、鲢、鳙、草鱼消化道结构与食性的研究

本文通过组织解剖与切片法测量了鲤、草、鲢、鳙的相关生物学参数,系统观察了四种鱼的消化道肌肉层、肠绒毛以及粘液细胞的显微结构及分布特点,探究其与食性的相互关系。结果显示：鲤比肠长1.0,环行肌较纵行肌发达,肠绒毛丰富,粘液细胞均匀分布在整个消化道中;草鱼比肠长2.13,消化道组织结构均一,肌肉壁中纵行肌所占比例高,粘液细胞体积小,分散于整个消化道中,肠绒毛极发达;鲢比肠长8.49,环行肌极发达,粘液细胞集中于消化道前段,肠绒毛由前至后逐渐呈短粗状;鳙比肠长4.58,环行肌极为发达,粘液细胞数量多且均匀地分布于消化道前、中段,肠绒毛较为稀疏短粗,粘液细胞较少。这四种鱼消化道的组织结构特征与各自食性密切相关。     

Effects of glycinin on growth performance,immunity and antioxidant capacity in juvenile golden crucian carp,Cyprinus carpio × Carassius auratus

This study was designed to examine the effects of glycinin on growth, digestive ability, immune responses, antioxidant capacity and gene expression levels of golden crucian carp. Golden crucian carp were fed diets containing glycinin at 0, 30, 60, 90 and 120 g/kg, respectively, for 8 weeks. Body weight, weight gain percentage, specific growth rate and feed efficiency ratio were negatively related to the content of glycinin in diet. Activities of protease, acid phosphatase, alkaline phosphatase, lysozyme in hepatopancreas, and activities of catalase, glutathione peroxidase, superoxide dismutase, and total antioxidant capacity in the proximal intestine, mid intestine, distal intestine and hepatopancreas were negatively related to the content of glycinin in diet, whereas malondialdehyde in proximal intestine, mid intestine, distal intestine and hepatopancreas increased directly with the content of glycinin in diet. Furthermore, the relative expressions of TNF‐α and IL‐1β in proximal intestine, mid intestine and distal intestine increased directly with the content of glycinin in diet, whereas the relative expressions of TNF‐α and IL‐1β in hepatopancreas were negatively related to the content of glycinin in diets. The relative expressions of IL‐10 in proximal intestine, mid intestine, distal intestine and hepatopancreas all were negatively related to the content of glycinin in diets. In conclusion, reductions in growth, immunity and antioxidant capacity, intestine inflammation with dysfunction of digestive system occurred in golden crucian carp that fed a diet containing glycinin at 30 g/kg or higher after 8 weeks.     

The effects of dietary inulin on growth performances,survival and digestive enzyme activities of common carp (Cyprinus carpio) fry

This study was conducted to investigate the effects of dietary inulin on growth performance, diet utilization, survival rate, carcass composition and digestive enzymes activities (amylase, lipase and protease) of carp (Cyprinus carpio) fry (0.55 ± 0.02 g). After acclimation, fish were allocated into 9 tanks (40 fish per tank) and triplicate fish groups were fed, control diet (0 g) or diets containing 5 g and 10 g inulin kg^?1 for 7 weeks. No significant effect on growth performance and diet utilization of fish fed inulin compared with the control group was observed. However, supplementation of inulin significantly increased survival rate and carcass lipid content, while carcass protein content significantly decreased. Dietary inulin had no significant effects on digestive lipase, protease and amylase activities.     

鲤鱼血清蛋白,血红蛋白,LDH和EST同工酶的电泳分析

采用聚丙烯酰胺凝胶电泳技术,分析了鲤鱼性成熟期雌雄个体的血清蛋白、血清脂蛋白和血红蛋白,结果是雌雄个体血清蛋白电泳图谱有差异,雌性(14条)比雄性(13条)多1条蛋白带;血清脂蛋白和血红蛋白无性别差异。同时,测定了鲤鱼血清、肝胰脏、心脏、背部肌肉和眼晶体的乳酸脱氢酶(LDH)同工酶和酯酶(EST)同工酶,LDH具有组织特异性,除在肝胰脏分离出9条酶带外,在血清中也分离出8条酶带,表明有LDH—c基因的作用,EST同工酶表现出明显的多态性,表明在鲤鱼各组织中有多个基因座位起作用。     

“全红”体色瓯江彩鲤MHC-DAB基因多态性及其与鱼体抗病力关系的分析

利用一对特异性引物DABF和DABR,分别从21尾感病和16尾抗病“全红”体色瓯江彩鲤的cDNA中,扩增出长度为624 bp的MHC-DAB基因片段。共测序185个有效克隆,获得76个不同的核苷酸序列。MHC-DAB基因片段包括第1~4外显子,分别编码信号肽、β1和β2结构域及跨膜区。长度为276 bp的β1结构域有144个核苷酸变异位点(52.17%)和70个氨基酸变异位点(76.07%),而长度为282 bp的β2结构域只有98个核苷酸变异位点(34.75%)和50个氨基酸变异位点(53.19%),显示β1结构域的变异要明显大于β2结构域。在β1结构域的24个抗原结合位点(PBR)上,有23个发生了变异。β1结构域的PBR、非抗原结合位点(non-PBR)及β2结构域的ω值(ω=d_N/d_S)分别为1.367、0.886和0.754,表明“全红”体色瓯江彩鲤MHC-DAB基因(特别是β1结构域)在进化过程中受到正向选择作用。在所得的20个不同的等位基因中,基因DAB3*15在抗病群体中出现的频率(13.75%)显著高于感病群体中出现的频率(3.81%)(P<0.05);基因DAB3*09和DAB3*10的出现频率都为3.75%(P<0.05),仅存在于抗病群体中;而基因DAB3*02和DAB3*14的出现频率分别为7.62%(P<0.05)和8.57%(P<0.01),仅存在于感病群体中。     

Application of Azotobacter enhances pond productivity and fish biomass in still water ponds

Two strains (derepressed-nitrogen fixing, Mac-27 and phosphate solubilizing, PS-21) of Azotobacter chroococcum were inoculated in fish culture ponds, singly and in combination with inorganic fertilizers (urea, single superphosphate–SSP). Physico-chemical parameters of pond waters, plankton production and fish biomass were studied. Inoculation of A. chroococcum (Mac-27) enhanced nitrogenase activity and rate of nitrogen fixation. A slight reduction in nitrogen fixation and nitrogenase activity was noticed when urea at 96 kg ha–1 y–1 was mixed with the biofertilizer (Mac-27). Inoculation of PS-21 enhanced phosphate solubilization, but Kjeldahl-nitrogen concentration values remained low in comparison with controls. On the other hand, inoculation of Azotobacter (either strain) enhanced the accumulation of ammonium-N, nitrite-N and nitrate-N. A significant (p < 0.05) reduction in dissolved oxygen (DO) concentration also took place when Azotobacter (both Mac-27 and PS-21) was inoculated in fish ponds. However, when used along with inorganic fertilizers, the reduction was not significant. The pH values were only slightly lowered when the phosphate-solubilizing strain (PS-21) of Azotobacter was inoculated. Inoculation of biofertilizer enhanced plankton production, net primary productivity and fish biomass. However, highest values in most of these parameters were noticed only in ponds that were treated with the higher doses of inorganic fertilizers (urea 192 kg and SSP 1500 kg ha–1 y–1). © Rapid Science Ltd. 1998     

鲤细胞因子多克隆抗体的制备及检测

为从蛋白质水平研究细胞因子在鲤体内免疫应答过程中的合成变化,本研究采用PCR技术克隆TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β基因含有部分抗原决定簇的片段,引入双酶切位点Bam HⅠ和HindⅢ后连接至p ET-32a/21a,构建相应的表达载体,制备多克隆抗体。采用ELISA检测抗体效价,并以此作为实验工具,检测经嗜水气单胞菌感染后鲤血清中炎性细胞因子的合成变化。结果显示,基因TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β融合蛋白分子量分别约为31.8、31.7、35.3、32.5、18.0和33.6 ku;抗体效价达到2.4×106;在病原菌感染后的不同阶段,促炎细胞因子TNF-α、IL-1β、IL-6、IL-12和抗炎细胞因子IL-10、TGF-β呈现出不同的合成变化。研究表明,制备的抗体具有较高的效价、亲和力和特异性,可用于鲤细胞因子的定量研究,该抗体的获得为鲤免疫应答与细胞因子合成的系统研究奠定了基础。同时,获取的鲤细胞因子TNF-α、IL-1β、IL-6、IL-12、IL-10和TGF-β的抗体亦可用于其他鱼类细胞因子蛋白质水平的定量研究。     

Effects of dietary medium-chain triacylglycerols (tricaprylin and tricaproin) and phospholipid supply on survival, growth and lipid metabolism in common carp (Cyprinus carpio L.) larvae

The present study investigated the interaction of dietary medium-chain fatty acids (MCFA) and phospholipids (PL) on survival, growth and lipid metabolism in common carp larvae. Nine diets based on casein and dextrin and with a variable lipid part were tested in triplicate for 22 days post first feeding. The 3×3 design consisted of three triacylglycerols (3% of diet) combined with three different lipid supplements. Tested triacylglycerols were triolein (TOL), tricaprylin (TC8) and tricaproin (TC6), and lipid supplements were 2% soybean oil (low-fat diets without PL), 2% soybean lecithin (low-fat diets with 2% PL) or both 2% soybean lecithin and 6% TOL (high-fat diets with 2% PL).

In the first step, both TC6 and TC8 resulted in improved survival and growth rates compared to TOL, irrespective of the PL supply. In the second step, TC8 decreased survival and growth rates, whereas the difference between TC6 and TOL became less. Histological signs of impaired intestinal absorption of neutral lipids were evidenced in larvae fed TOL without PL and also in high-fat diets with 2% PL. The latter diets also resulted in poorer growth rates compared to low-fat diets with 2% PL. These results suggest that the quantitative PL requirement of larvae increases as the dietary level of long-chain triacylglycerols increases. Larvae fed TC6 or TC8 showed enlarged liver and hepatocyte volume and a decreased level of body neutral lipids. Based on β-hydroxybutyrate (β-HBA) measurements in whole larvae, TC8 was found to be more ketogenic than TC6. TC6 and TC8 affected differently the fatty acid profile of larval body neutral lipids. TC6 did not induce the appearance of MCFA, whereas TC8 feeding resulted in a low level of 8:0 and relatively high levels of 10:0 (3.8% of total fatty acids). Neither 8:0 nor 10:0 were found in larval polar lipids.

This study confirmed the essentiality of PL in common carp larval diets and underlines differences in the utilization of TC6 and TC8, which both initially stimulate growth during the first week, but only temporarily in the case of TC8.