谷氨酰胺对黄颡鱼幼鱼抗氧化能力和非特异性免疫力的影响

本试验旨在研究谷氨酰胺对黄颡鱼幼鱼抗氧化能力及非特异性免疫力的影响。选取平均体重为(2.49±0.04)g的黄颡鱼幼鱼240尾,随机分为4组,每组3个重复,每个重复20尾。4组试验鱼分别饲喂谷氨酰胺添加量为0(对照)、0.1%、0.2%、0.4%的等氮等能试验饲料。试验期为10周。结果表明:饲料中添加0.2%的谷氨酰胺可以显著提高黄颡鱼幼鱼的血清总蛋白、球蛋白、甘油三酯、高密度脂蛋白和低密度脂蛋白含量(P0.05);各组黄颡鱼幼鱼的血清谷丙转氨酶、谷草转氨酶和碱性磷酸酶活性差异不显著(P0.05);饲料中添加0.1%的谷氨酰胺可以显著提高黄颡鱼幼鱼肝脏中过氧化氢酶、谷胱甘肽过氧化物酶及肌肉中超氧化物歧化酶、谷胱甘肽过氧化物酶的活性(P0.05);随着饲料中谷氨酰胺添加量的增加,头肾巨噬细胞的吞噬指数呈升高趋势,且0.2%和0.4%组较对照组显著升高(P0.05),但0.2%和0.4%组间差异不显著(P0.05)。由此可见,饲料中添加0.1%~0.2%的谷氨酰胺能够提高机体的抗氧化能力和非特异性免疫力。     

黄颡鱼毒棘结构研究

黄颡鱼（Pelteobagrus fulvidraco）是较为典型的淡水刺毒鱼类,本研究结合宏观毒棘骨骼标本制作和微观组织切片等方法,深入探究黄颡鱼的毒棘结构。研究结果表明,黄颡鱼的胸鳍和背鳍均具有尖锐的倒刺棘骨,棘骨外包围着皮膜,皮膜拥有能分泌毒液的毒腺细胞,它们共同组成了黄颡鱼毒棘装置。毒棘骨骼宏观结构表明,黄颡鱼的背鳍和胸鳍毒棘均具有贯穿全棘骨的中央管和包围棘骨的皮膜;背鳍棘骨细长、锥形,且轻微拱起,顶部骤尖,后缘为弱锯齿;胸鳍棘骨前后缘均有锯齿,但后缘为强锯齿。背鳍和胸鳍毒棘基部结构的髁突形状、大小、位置全然不同。微观结构显示,皮膜中的毒腺细胞聚集成层,位于鳞状上皮与色素层之间,未见导管与之相连,背鳍和胸鳍毒棘的分支性骨管中均未发现毒腺细胞。胸鳍和背鳍锁紧装置可让毒棘保持倒伏或者直立的锁定状态,降低猎物挣脱几率。本研究可深入了解黄颡鱼的毒棘结构和特征,增强人们对刺毒鱼类的认知,同时为淡水刺毒鱼类深入研究奠定基础。     

黄颡鱼不同组织5 种同工酶的分析

为研究黄颡鱼同工酶系统的组织特异性,分析黄颡鱼的遗传多样性并为品种改良提供有价值的生化遗传指标。采用聚丙烯酰胺垂直凝胶电泳法,对黄颡鱼的血,腮,脑,肝,心,胃,肾,性腺,肌肉,肠,头肾,脾和膀胱共13种新鲜组织中的苹果酸脱氢酶（MDH）、苹果酸酶（ME）、超氧化物歧化酶（SOD）、酯酶（EST）、葡萄糖醛酸酐酶（GD）共5种同工酶进行了分析。结果表明,MDH在脑、肾和头肾中分别具有特异性酶带;ME只在肝组织检测到酶带;SOD在肝、肠、脑和肌肉组织分别具有特异性酶带,EST在肌肉组织具有特异性酶带;GD在腮和脑组织中具有特异性酶带。说明黄颡鱼5种同工酶系统具有明显的组织特异性。     

Ontogeny of the stomach in yellow catfish (Pelteobagrus fulvidraco): detection and quantifictation of pepsinogen and H+/K+‐ATPase gene expression

Yellow catfish (Pelteobagrus fulvidraco) is an important commercial species with high aquaculture potential in China. To better understand the process of digestive functioning of gastric gland development during the larval from 1 dph (day post‐hatching) to 30 dph, real‐time PCR was used to detect and quantify the pepsinogen and H⁺/K⁺‐ATPase gene expression in P. fulvidraco. These data were also compared with the adult situation. The results showed that the expression of pepsinogen and H⁺/K⁺‐ATPase genes in P. fulvidraco larvae both started at 1 dph, though the expression level was very low until 3 dph. The quantification of pepsinogen gene expression increased significantly from 4 to 8 dph, increased fluctuantly from 8 to 23 dph and rose sharply from 23 to 30 dph. In comparison with adult fish, there were no significant differences with larvae at 5 and 23 dph. However, data of 10 and 30 dph larvae were obviously higher than those of adult group. H⁺/K⁺‐ATPase gene expression increased linearly from 1 to 30 dph. However, it was significantly lower than that of adult. The results show that P. fulvidraco larvae have an earlier functional stomach, though the function of the stomach is still not perfect. There is a gradual acidification environment within the stomach during the P. fulvidraco larvae development. Based on these results, we suggest that the weaning time for P. fulvidraco larvae would be much better after 23 dph.     

鮰爱德华菌单克隆抗体的制备及其在黄颡鱼“红头病”研究中的应用

以黄颡鱼Pelteobagrus fulvidraco “红头病”病原---鮰爱德华菌 Edwardsiella ictaluri A86为抗原,利用杂交瘤技术制备了鮰爱德华菌黄颡鱼分离株的菌体单抗5D11、1B8、1G7、3G8、5A9、3H8,其效价分别为1：1280、1：1280、1：2560、1：320、1：160和1：80。结果表明：6株单抗与11株鮰爱德华菌黄颡鱼分离株均可以结合,与迟缓爱德华菌、气单胞菌、大肠杆菌、链球菌、弧菌等10株参考菌株均无交叉反应,单抗5D11和1B8可与鮰爱德华菌参考菌株结合。应用免疫荧光和免疫组织化学法检测了鮰爱德华菌对黄颡鱼的侵染规律。结果表明：黄颡鱼经人工注射鮰爱德华菌A86感染6 h后,在其肝、肾、脾、心、肠和胃中检测到了阳性细菌,其中脾脏中最多,心脏中最少;感染24 h后,脑中检测到阳性细菌;30 h后,鳃中显示有阳性信号;54 h后,在眼的外边缘检测到少量阳性细菌。研究结果可为进一步研究鮰爱德华菌在黄颡鱼体内的致病机理提供技术手段及基础,对揭示黄颡鱼“红头病”的发病机制具有重要意义。     

冷、热休克法诱导黄颡鱼三倍体的比较研究

分别采用冷、热休克抑制第二极体释放的方法诱导黄颡鱼三倍体。结果表明,在卵受精后2min,5℃处理20min,胚胎时期的三倍体率达70%左右,孵化率50%左右,幼鱼时期三倍体(含嵌合体)的检出率为25%,此条件为冷休克处理的优化参数;在卵受精后2min,40℃处理2min,胚胎时期的三倍体诱导率达58%,孵化率为39%,幼鱼时期三倍体(含嵌合体)的检出率为40%,此条件为热休克处理的优化参数。正交分析得出,冷休克条件下起始休克时间是原肠期三倍化率和孵化率的重要影响因子,温度对畸形个体的产生有重要影响;热休克条件下,参考三倍体率、畸形率、孵化期相对存活率三者而言,休克温度均是重要因素。比较观察到冷休克处理组的胚胎受损情况严重,后期的成活率较热休克处理组要低,总体诱导效果逊于热休克处理组。     

嘉陵江野生瓦氏黄颡鱼人工繁殖技术的研究

对嘉陵江野生瓦氏黄颡鱼进行了人工繁殖和苗种培育试验。结果表明5月中旬水温达到24℃左右时为开展人工繁殖的最佳时间;催产药物种类和剂量以HCG（400IU）＋LHRH-A2（3μg）＋DOM（0．25mg）／尾效果最好;使用编织袋做成的鱼巢孵化率最高;用蛋黄、浮游动物和水蚯蚓作为饵料培养仔鱼效果良好。     

拟态弧菌感染黄颡鱼的动态病理损伤及病原分布研究

以1.0×106CFU·m L~(-1)拟态弧菌(Vibrio mimicus)浸浴感染黄颡鱼(Pelteobagrus fulvidraco),于第0、第4、第8、第16、第24、第36、第48、第60、第72小时剖杀取样,研究病理损伤及病原菌的体内动态分布。结果显示,皮肤肌肉、鳃、肠道出现病变最早,其中皮肤肌肉损伤最严重。第16小时表皮变性、坏死,鳃上皮肿胀,肠绒毛水肿;第24~第48小时表皮坏死、脱落,真皮严重出血,肌纤维变性、坏死,鳃出血,上皮局部坏死,肠上皮变性与灶性坏死;第48~第72小时皮肤肌肉坏死更严重,形成溃疡灶,鳃灶性坏死。肾、肝、脾与心36 h后相继出现不同程度的淤血、出血,变性和灶性坏死。q PCR检测发现,第4小时即在鳃、肠、皮肤肌肉检出病菌,细菌含量分别为1.3×102CFU·mg~(-1)、2.6×102CFU·mg~(-1)、4.7×102CFU·mg~(-1);鳃与皮肤肌肉中菌量随时间延长而增加,第72小时皮肤肌肉中菌量达到4.5×107CFU·mg~(-1);第24小时后相继在肾、肝、脾、心检出病菌,菌量介于1.9×10~4.7×102CFU·mg~(-1)之间。结果表明皮肤、鳃和肠道是病菌的入侵位点,并在体内多组织、器官分布。     

饲料磷含量对黄颡鱼幼鱼生长、生化组成及血清生化指标的影响

在黄颡鱼商业饲料基础上通过减少Ca(H2PO4)2的添加量,制成3种不同磷含量(以Ca(H2PO4)2计）的饲料(25kg/t简称25P、15kg/t简称15P、5kg/t简称5P)进行生长试验,测定不同磷含量对黄颡鱼生长、生化组成和血清生化指标的影响.每种饲料对应3缸重复,每缸15尾鱼（初始平均体质量3.2g）,进行...     

Isolation and Pathogenicity Analyses onYersinia enterocoliticafrom Pelteobagrus vachelli

Yersinia enterocolitica is an important zoonotic pathogen that can induce disease outbreaks in a wide host range. Strain YER6022 was isolated from Pelteobagrus vachelli and identified using bacterial morphology and 16 SrDNA sequence analysis. Five virulence factors were detected, then artificial infection experiment and histopathological method were carried out. These results showed that strain YER6022 was one of Y. enterocolitica family members. In addition, ail, ystb, vir F, yad A and HPIint were dectected. In artificial infection experiment, with 80% mortality and 100% morbidity, injected Pelteobagrus vachellis showed red swollen of the anus, abdomen swelling and fim bleeding. There existed serious hyperaemia and edema in kidney, spleen, intestine and liver at the light microscope. Ultrastructural observation indicated that mitochondria of the liver, kidney, spleen and intestine swelled and mitochondrial cristae broke. The data had further shed light on its pathogenicity in Pelteobagrus vachelli. It would benefit for further studies on pathogenesis of Pelteobagrus vachelli infected with Y. enterocolitica.