Generation of mouse monoclonal antibodies specific to tilapia immunoglobulin using fish immunoglobulin/BSA complex for monitoring of the immune response in Nile tilapia Oreochromis niloticus

The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti‐BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer, as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.     

饲料添加芦丁对吉富罗非鱼血清炎症因子的影响

为了摸索饲料中芦丁的最适合添加浓度,在饲料中分别添加芦丁0、0.1、0.3 g/kg(分别为对照、低浓度组、高浓度组)饲喂吉富罗非鱼幼鱼,于第15、30、45天检测血清IgM,抗炎因子和促炎因子的含量。结果表明芦丁添加组吉富罗非鱼血清IgM(除45天低浓度组),抗炎症因子IFN-γ(除45天高浓度组)、IL-10(30天)含量显著升高,促炎症因子TNFα、IL-1β和IL-8(30天,45天低浓度组)含量显著降低。本试验证实一定浓度的芦丁对吉富罗非鱼幼鱼具有炎症效应。     

投喂频率对尼罗系吉富罗非鱼幼鱼胃排空、生长性能和体组成的影响

通过胃排空试验与养殖试验研究了不同投喂频率对尼罗系吉富罗非鱼幼鱼的胃排空、生长性能以及体组成的影响.在试验开始时,观测尼罗系吉富罗非鱼幼鱼的胃内饲料排空情况,胃排空试验结果表明,胃排空率的最佳描述为平方根函数,胃内饲料在饱食投喂后15 h左右完全排空,达到投喂前水平,80％胃排空为9h,也就是投喂后大约9h恢复食欲.360尾试验鱼(初始体质量3.72 g)以不同的投喂频率(1d4次、1d3次、1d2次、2d4次、2d3次、2d2次)分组,每组设立3个平行组,随机养殖于18个网箱中,每箱养殖20尾鱼,按饱食量投饲膨化饲料.养殖期为6周.尼罗系吉富罗非鱼幼鱼在投喂频率为1d4次、1d3次和1d2次时特定生长率和饲料效率显著高于投喂频率为2d4次、2d3次和2d2次时(P＜0.05)；投喂频率为1d2次、2d4次、2d3次和2d2次时其摄食量显著低于1d4次和1d3次时(P＜0.05).随着投喂频率降低,鱼体水分含量逐步上升,脂肪和蛋白质含量逐步下降,其中1d4次、1d3次组鱼与2d2次组鱼有显著性差异(P＜0.05).各投喂频率组间的肝体指数无显著性差异(P＞0.05).3.7～48.0 g尼罗系吉富罗非鱼幼鱼的适宜投喂频率为1d2次,较2d2次、2d3次和2d4次时明显提高了生长速度和饲料效率,较1d3次、1d4次摄食量显著降低.2种试验结果较为一致.     

The effect of nearly closed RAS on the feed intake and growth of Nile tilapia (Oreochromis niloticus), African catfish (Clarias gariepinus) and European eel (Anguilla anguilla)

One of the challenges that Recirculating Aquaculture Systems (RAS) are still facing is the risk that in RAS fish grow less than in flow-through systems due to the accumulation of substances originating from feed, fish or bacteria associated with the water re-use. The present study investigated whether RAS with high and low accumulation levels of these substances affect feed intake and growth of Nile tilapia Oreochromis niloticus, African catfish Clarias gariepinus, and European eel Anguilla Anguilla. One-hundred and twenty individuals of each species were used (start body weights: Nile tilapia 264.8 ± 8.3 g; African catfish 253.2 ± 2.1 g and European eel 66.6 ± 1.3 g). For a period of 39 days, growth and feed intake were compared between high and low accumulation RAS. HIGH accumulation RAS was designed for maximal accumulation of substances in the water by operating the system at nearly-closed conditions (30 L/kg feed/d), using mature biofilters and high feed loads; and (2) LOW accumulation RAS was designed to be a proxy for flow-through systems by operating at high water exchange rates (1500 L/kg feed/d), new biofilters and low feed load. HIGH accumulation RAS induced a reduction in feed intake (42%) and growth (83%) of Nile tilapia, as compared to systems that are a proxy for flow-through conditions. This effect was not observed in European eel and African catfish. The cause of this reduced feed intake and growth rate of Nile tilapia is still unclear and should be addressed in further studies.     

半滑舌鳎溃疡病原杀鲑气单胞菌的分离鉴定与药敏试验

半滑舌鳎(Cynoglossus semilaevis Günther)是广受我国消费者认可的名优鱼类,具有很高的营养价值和经济价值,然而,随着工厂化养殖模式的不断发展,疾病成为制约半滑舌鳎养殖业健康发展的关键因素,其中,体表溃疡病为其常见病症。从严重溃疡病半滑舌鳎体内分离到1株优势菌株HX0416,该菌可在TCBS培养基上生长,并可在绵羊血平板上形成清晰的β-溶血环;经过16S rRNA和rpo D基因序列同源性分析并结合生化特性鉴定为杀鲑气单胞菌(Aeromonas salmonicida)。健康半滑舌鳎幼鱼感染试验表明,该菌对半滑舌鳎具有较强致病性,感染72 h(1.04×106cfu·m L-1)对攻毒鱼的致死率达91.7%。药敏试验结果显示,该菌对环丙沙星、恩诺沙星、诺氟沙星等喹诺酮类和头孢曲松、头孢吡肟等头孢菌素类药物比较敏感,而对硫酸新霉素、土霉素等氨基糖苷类和四环素类药物多重耐药,尤其是对磺胺甲噁唑、氨苄西林等磺胺类和β-内酰胺类药物严重耐药。     

三黄连散预防罗非鱼链球菌病临床应用研究

在开展三黄连散对人工感染罗非鱼无乳链球菌病预防效果研究的基础上,进一步验证以每千克饲料中添加13.2 g的三黄连散为推荐剂量,在生产中大面积应用的临床效果及安全性。设立三黄连散试验组、中药对照组和空白对照组3个组,每组300尾罗非鱼,连续用药7 d后,每组随机取30尾,测定增重率、器官指数、血液生理生化指标和肝脏抗氧化指标。用药后第8天,采用同居感染的方法感染鱼群,于感染后第14天计算保护率。结果表明,给药7 d后,与空白对照组相比,三黄连散试验组罗非鱼的增重率明显提高,器官指数和多项肝脏抗氧化指标均明显升高,血液生理生化指标总体无明显异常,预防无乳链球菌感染的保护率为43.67%。综上提示,三黄连散能够提高罗非鱼机体抵抗力,降低无乳链球菌感染罗非鱼的死亡率。     

Modeling and implementing the use of aeration to increase water temperature and dissolved oxygen in greenhouse aquaculture cages

In this study, an unique bamboo floating cage was built in order to heat the water used for rearing Nile tilapia (Oreochromis mossambicus x O. niloticus) and climbing perch (Anabas testudineus, Bloch) in Northern Thailand during the colder rainy and winter seasons. This special greenhouse fish cage (SGFC) was covered with polyethylene to heat the surface air with solar energy, and insulated to prevent heat loss. The water temperature at depths of 0.1, 0.3 and 0.8 m was modeled and measured. In various trials, the cages were aerated with and without heated air from the surface. It was found that the thermal efficiency of the SGFC integrated with hot air aeration at a water depth of 0.80 m was 32.65 %. Throughout the day, the average water temperature was 31.82 ± 0.68 °C, which was 0.76 °C higher than the water temperature outside the experiment, and the amount of oxygen in the water of 11.35 ± 0.13 mg/l. The SGFC can produce one extra fish harvest per year when compared with regular fish cages. Development equations to describe the thermal model were used for predicting the water temperature in the SGFC and thus validating the model. Moreover, the economic analysis of the SGFC found that the payback period was one year, seven months and fourteen days.     

Occurrence of multiple antibiotic resistance and genotypic characterization in Edwardsiella tarda isolated from cage‐cultured hybrid red tilapia (Oreochromis sp.) in the Ping River,Northern Thailand

Edwardsiella tarda is a pathogen that causes edwardsiellosis in aquatic animals. The emergence of multiple antibiotic‐resistant strains makes antibiotic treatment difficult. This study aimed to investigate the antibiotic susceptibility patterns and the genotypic characterization of E. tarda isolated from cage‐cultured red tilapia in Thailand. A total of 30 isolates were identified as E. tarda using biochemical and molecular analysis. The disc diffusion method for testing antibiotic susceptibility showed all the isolates were resistant to colistin sulphate and oxolinic acid. High levels of resistance to amoxicillin, ampicillin, ceftazidime, oxytetracycline and sulphamethoxazole/trimethoprim were observed as well. The multiple antibiotic resistance index ranged from 0.25 to 0.92, indicating that these isolates had been exposed to high risk sources of contamination where antibiotics were commonly used. All the isolates carried the bla_TEM gene based on polymerase chain reaction (PCR). The tetA and sul3 genes were detected in 90% (27/30) and 26.7% (8/30) of the isolates respectively. Nine different genetic groups of isolates were obtained using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC‐PCR). A correlation between genetic types and multiple antibiotic‐resistant patterns was found. These results highlight the potential risks of multiple antibiotic‐resistant isolates for humans and the environment.     

两种油脂对小鼠皮肤创伤修复作用研究

本研究旨在观察阿勒泰大尾羊尾脂油脂对损伤皮肤是否具有修复作用，并与牛油脂、市售商品做比较。研究选择40 只体重在31 ± 1 g 之间的昆明小鼠，雌雄各半，通过构建小鼠皮肤损伤模型，将小鼠分为4 组（羊尾油组、牛油组、市售商品组和空白组）进行试验观察。小鼠皮肤伤口愈合良好，雌鼠的创面愈合能力强于雄鼠，雌鼠伤口愈合速度：羊尾油组＞牛油组＞市售商品组＞空白对照组。雄鼠伤口愈合速度：羊尾油组＞市售商品组＞牛油组＞空白对照组。皮肤组织病理学结果显示，涂抹羊尾脂、牛油脂、市售商品与空白对照处皮肤相比，创伤修复组织中脂肪细胞、胶原纤维以及其他细胞的数量、排列分布均有差异。羊尾油和牛油均有修复皮肤创伤的作用，且羊尾油修复作用更强，羊尾油可通过促进胶原纤维的生成加速皮肤修复。