壳聚糖和益生菌对暗纹东方鲀抗病力和免疫功能的影响

为了解壳聚糖和益生菌刺激和增强免疫应答的可能作用,本试验在基础饲料中添加0.2%、0.5%、1.0%的壳聚糖,0.1%、0.2%、0.4%的益生菌,甘露聚糖与益生菌混合物,壳聚糖与益生菌的混合物,在室内水泥池中喂养暗纹东方鲀(3.15±0.05)g 2个月,根据抗菌活力筛选出免疫增强饲料组(0.2%壳聚糖,0.1%、益生菌,甘露聚糖与益生菌混合物,壳聚糖与益生菌混合物)测定免疫功能。结果表明,甘露聚糖与益生菌混合物仅提高头肾T淋巴细胞转化,壳聚糖与益生菌混合物显著增强头肾T、B淋巴细胞转化和脾脏B淋巴细胞体外培养时分泌的IgM。筛选出的四种免疫增强饲料均能使头肾B淋巴细胞体外培养时分泌的IgM显著增加。无论是脾脏B淋巴细胞还是头肾B淋巴细胞,体外培养时所分泌的IFN-α都不同程度地受抑制。     

浅析维生素C、E混合物以及免疫多糖对暗纹东方鲀生长及抗病力的影响

免疫增强剂的研究已成为当前鱼病学研究和新鱼药开发的一个热点。作为一种新型的绿色渔药,免疫增强剂以其种类多、应用广、使用方便等优点,日渐受到人们的青睐。研究表明,免疫增强剂在鱼类疾病控制上具有十分重要的意义。因此,免疫增强剂添加在暗纹东方鲀饲料中的应用将成为生态、绿色、健康养殖的趋势。本试验主要研究了维生素C、维生素E以及免疫多糖等免疫增强剂在暗纹东方鲀上的应用。     

玉米蛋白粉替代鱼粉对暗纹东方鲀溶菌酶活性及c型溶菌酶mRNA表达的影响

以初重为(41.26±1.09)g的暗纹东方鲀为实验鱼,配制5种玉米蛋白粉使用量为0%、5%、10%、15%、20%的实验饲料,玉米蛋白粉对饲料中鱼粉的替代水平分别为0%、7.4%、14.8%、22.2%和29.6%。用实验饲料饲养暗纹东方鲀60d,研究玉米蛋白粉替代鱼粉对暗纹东方鲀肝胰脏、头肾和脾脏组织的溶菌酶活性及其c-型溶菌酶mRNA相对表达量的影响。结果表明,(1)暗纹东方鲀肝胰脏、头肾和脾脏的溶菌酶比活力分别为9.14、42.12和40.49U/mgprot,头肾和脾脏组织中c-型溶菌酶mRNA相对表达量分别是肝胰脏的3.76和3.24倍。(2)玉米蛋白粉替代鱼粉显著影响暗纹东方鲀溶菌酶比活力。5%和10%组的肝胰脏和头肾组织溶菌酶比活力显著高于对照组(P<0.05),而15%和20%组肝胰脏和头肾组织溶菌酶比活力则显著低于对照组(P<0.05);5%和10%组的脾脏组织溶菌酶活力与对照组无显著性差异(P>0.05),而15%和20%组显著低于对照组和5%组(P<0.05)。(3)玉米蛋白粉对暗纹东方鲀c型溶菌酶基因mRNA相对表达量有显著影响(P<0.05)。肝胰脏组织中5%和10%组表达量显著高...     

暗纹东方鲀血细胞发生的观察

通过对暗纹东方鲀(Takifugu obscurus)血液涂片及头肾、体肾、脾脏和肝脏四种脏器印片的光镜观察,发现血细胞的发育大致经过三个阶段,即原始阶段、幼稚阶段、成熟阶段。实验对不同发育阶段的红血细胞、淋巴细胞、单核细胞、粒细胞进行了观察和测量,并对暗纹东方鲀血细胞的发生过程做了初步探讨。实验结果表明暗纹东方鲀血细胞的发育主要在头肾和体肾,肝脏印片未观察到原始造血细胞,提示肝脏可能不是暗纹东方鲀的造血器官。     

暗纹东方鲀江水漂养高效试验

为了提高暗纹东方鲀的养殖效益和品质,我们从2012年4月至10月,在泰兴市长江渔场进行暗纹东方鲀江水漂养试验。试验共使用3口鱼池,9亩水面,每口鱼池3亩。每亩放养暗纹东方鲀1000尾,搭配鲢鱼200尾,投喂粗蛋白45%粉状饲料和蚌肉、小鱼等鲜活饵料,取得了良好的效益。现将试验情况总结如下:     

一种复合免疫增强剂对大菱鲆和血鹦鹉鱼酶活性及抗病力的影响

在饲料中对自制复合免疫增强剂设计了四个添加剂量(投喂免疫增强剂浓度0‰、投喂免疫增强剂浓度1‰、投喂免疫增强剂浓度2‰、投喂免疫增强剂浓度3‰)分别投喂大菱鲆和血鹦鹉鱼。30d后测定各组试验鱼血清的溶菌酶活性、碱性磷酸酶活性和总超氧化物歧化酶活性,并分别进行攻毒试验。结果表明投喂免疫增强剂浓度1‰、2‰的大菱鲆的溶菌酶活性、碱性磷酸酶活性和总超氧化物歧化酶活性皆显著高于对照组(P0.05);三个实验组鱼的人工感染死亡率分别为50%、45%和100%,方差分析表明两个实验组皆显著低于对照组(P0.05)。投喂血鹦鹉鱼试验各测定酶指标都是2‰组最高,并且显著高于对照组(P0.05),攻毒试验死亡率投喂投喂免疫增强剂浓度2‰组最低。因此可以证明投喂本复合制剂可以明显提高大菱鲆和血鹦鹉鱼的酶活性及抗应激能力,并且适宜投喂比例为2‰。     

几种免疫增强剂对草鱼SOD、CAT及AKP活性的影响

将草鱼放入网箱中,投喂在基础饲料中分别添加不同剂量的黄芪多糖、枸杞多糖和金丝桃素的饲料,研究其对草鱼血清中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及碱性磷酸酶(AKP)活性的影响。结果表明:投喂添加中药免疫增强剂的饲料60d后,与对照组相比,血清中CAT和AKP活性变化不明显(P0.05);血清中SOD活性显著高于对照组(P0.05),说明中药免疫增强剂对增强草鱼免疫活性具有一定的作用。     

壳聚糖投喂方式对草鱼抗饥饿胁迫能力的影响

在基础饲料中添加0.5%的壳聚糖,采用3种不同的投喂方式(方式A:连续投喂基础饲料,对照组;方式B:连续投喂添加0.5%壳聚糖的饲料,连续组;方式C:先投喂0.5%壳聚糖饲料再投喂基础饲料且每15天间隔投喂,不连续组)饲喂初始体重(19.46±0.04)g的草鱼60d后,对草鱼进行饥饿胁迫处理[各投喂方式分为投喂组(feeding,F)和饥饿组(starvation,S)],以生长、一氧化氮(nitrogenoxide,NO)含量和溶菌酶(lysozyme,LSZ)活性为指标考察壳聚糖不同投喂方式对草鱼抗饥饿胁迫处理的能力。结果显示,(1)连续组和不连续组60d时草鱼增长率和增重率显著高于对照组(P<0.05),15d饥饿处理后也呈现投喂组和饥饿组的增长率和增重率高于对照组的趋势(P>0.05);(2)对照组头肾、肝胰脏NO含量和血清、头肾溶菌酶活性饥饿组显著高于投喂组(P<0.05),连续组除头肾外NO含量和各组织溶菌酶活性饥饿组显著低于投喂组(P<0.05)或与投喂组无显著差异(P>0.05),不连续组除脾脏外NO含量和除肝胰脏外的溶菌酶活性饥饿组显著低于投喂组(P<0.05)或与投喂组无显著差异(P>0...     

免疫低聚糖对中国对虾的免疫效用研究

为证实免疫低聚糖(D-氨基葡聚寡糖,D-Amino-Oligosacellaride)对中国对虾(Penaeus chinensis)的免疫增强作用,进行了不同剂量的注射及饲料投喂实验.结果表明,注射免疫低聚糖可显著增强中国对虾酸性磷酸酶和溶茵酶等血清免疫指标,添加于饲料中投喂可以提高对虾成活率,降低饵料系数,且饲料中添加量≥3‰的效果较佳.     

褶皱臂尾轮虫对提高暗纹东方鲀鱼苗成活率的作用

利用褶皱臂尾轮虫和暗纹东方鲀鱼苗在5‰的咸淡水中都能够存活的特点,用面包酵母培育褶皱臂尾轮虫经海水小球藻强化培育后在5‰的盐度条件下投喂暗纹东方鲀鱼苗,结果表明,褶皱臂尾轮虫较之淡水轮虫可使暗纹东方鲀鱼成活率显著提高。     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.