Effect of different feeds on the growth,survival and reproductive performance of zebrafish,Danio rerio (Hamilton, 1822)

Present experiment was conducted to determine the effect of different feeds with varying protein levels on the growth, survival and reproductive performance of zebrafish, Danio rerio. The control diet (T1) was wild‐collected zooplankton from local fish ponds, while test diets with 350 g kg^?1 protein (T2), 400 g kg^?1 protein (T3) and 450 g kg^?1 protein (T4) were formulated and fed to fish for a period of 210 days. The significantly (P < 0.05) highest mean weight gain and specific growth rate were observed in T1, which were similar with T3 and T4. The significantly (P < 0.05) highest number of egg production per female and relative fecundity were found in T1, followed by T4 and T3 while T2 produced lowest number of eggs. No significant (P > 0.05) differences were observed in brood survival rate, fertilization and hatching rate among the dietary treatments. The highest (P < 0.05) fry survival rate was recorded in T1, followed by T3 and T4. Thus, it is suggested that control diet i.e. mixed zooplankton exhibited better growth, reproductive performance and fry survival rate. However, diet containing 400 g kg^?1 crude protein also gave comparable results in terms of growth, survival and reproductive performance of zebrafish.     

First report of Fusarium oxysporum species complex infection in zebrafish culturing system

Fusarium oxysporum species complex (FOSC) is a highly diverse fungus. Recently, F. oxysporum infection was identified from zebrafish (Danio rerio) culturing system in Korea. Initially, a rapid whitish smudge was appeared in the water with the fungal blooming on walls of fish tanks. Microscopic studies were conducted on fungal hyphae, colony pigmentation and chlamydospore formation and the presence of macro‐ and microspores confirmed that the isolated fungus as F. oxysporum. Furthermore, isolated F. oxysporum was confirmed by internal transcribed spacer sequencing which matched (100%) to nine F. oxysporum sequences available in GenBank. Experimental hypodermic injection of F. oxysporum into adult zebrafish showed the development of fungal mycelium and pathogenicity similar to signs observed. Histopathologic results revealed a presence of F. oxysporum hyphae in zebrafish muscle. Fusarium oxysporum growth was increased with sea salt in a concentration‐dependent manner. Antifungal susceptibility results revealed that F. oxysporum is resistant to copper sulphate (up to 200 μg mL^?1) and sensitive to nystatin (up to 40 μg mL^?1). This is the first report of FOSC from zebrafish culture system, suggesting it appears as an emerging pathogen, thus posing a significant risk on zebrafish facilities in the world.     

Effects of prebiotic mannan oligosaccharide on the growth,survival, and anxiety-like behaviors of zebrafish (Danio rerio)

A study was designed to investigate the possible positive prebiotic effects of mannan oligosaccharides (MOS) on growth and reduction of anxiety behaviors in zebrafish (Danio rerio). The experimental diets were supplemented with 0, 2, 4, 6, or 8 g MOS kg^?1 and were fed to juvenile fish (0.47 ± 0.017 g) in triplicates for six weeks. Significant differences were found in the final body weight, body weight intake, percent body weight intake, and specific growth rate among treatments. The regression analyses showed that the optimum MOS level was 4–4.1 g MOS kg^?1 diet. Fish fed 4 g MOS kg^?1 feed showed higher survival than the other treatments. Also, using the novel tank test, fish fed with 4 g MOS kg^?1 diet showed lower anxiety by swimming in the upper portion of the tank. The other behavior parameters remained unaffected by the prebiotic. The results indicated the beneficial effect of the prebiotic MOS on growth and partly on welfare of zebrafish juveniles.     

谷甾醇对斑马鱼生长发育和生殖的影响

为了研究谷甾醇(sitosterol)暴露对斑马鱼(Barchydanio rerio var)内分泌的干扰效应,通过不同浓度的谷甾醇(50、100、150、200μg/L)对雌性斑马鱼进行为期60 d的水浴暴露,同时设乙醇对照,分别于60 d后测定各组分雌性斑马鱼的生长指标,对部分组织进行切片观察,从组织病理学和毒理学方面评估谷甾醇暴露的毒性效应。结果表明:在200μg/L谷甾醇暴露下,斑马鱼全部死亡,其他各浓度处理与对照组相比,除臀鳍数随谷甾醇浓度的增加呈现倒"U"型趋势,以及肝重随谷甾醇浓度的增加而上升外,其他各生长指标均不同程度受到抑制,抑制作用与暴露浓度呈明显的剂量效应。此外,性腺指数(GSI)呈下降趋势,肝体指数(HSI)呈上升趋势,各组织切片也表现出明显的毒性反应。因此,可初步判断谷甾醇对斑马鱼的生长发育和繁殖系统会产生较大影响。     

甲氰菊酯对斑马鱼的毒性效应

选用模式生物斑马鱼(Brachydanio rerio)作为毒性实验动物,探索甲氰菊酯(fenpropathrin)对鱼类的毒性效应,为农业生产和环境保护提供参考数据。结果显示甲氰菊酯对斑马鱼成鱼96h LC50为3.38×10-3mg/kg,属于剧毒级,安全浓度为0.34×10-3mg/kg;较低浓度下甲氰菊酯对斑马鱼早期发育影响不显著,在较高浓度1、10mg/kg下,斑马鱼96h畸形率分别达到84%和90%,心包囊肿率分别为25%和88%,而致死效应不明显。亚致死剂量下甲氰菊酯对斑马鱼肝胰脏GPT、GOT、SOD活性有明显诱导效应,在高浓度和长时间暴露下诱导效应降低;肝胰脏中AChE活性显著地受到甲氰菊酯的抑制,抑制效应对毒物的浓度和染毒时间有良好的依赖性,是一种良好的环境污染检测生物学指标。     

Characterization of two microparticle types for delivery of food to altricial fish larvae

The characteristics of two microparticulate types were determined for delivery of nutrients to fish larvae. A yeast-based commercial diet for larval fish (Microfeast Plus L-10, Salt Creek, Inc., Salt Lake City, UT, USA) was encapsulated within cross-linked protein-walled capsules (CLPWC) or gelatin-alginate beads (GAB). Microparticle stability was determined by encapsulating a nontoxic red dye (Poly R 478) within CLPWC or GAB. The stabilities of both particle types suggested that high molecular weight, water-soluble nutrients could be delivered to freshwater fish larvae without major losses from leaching. Acceptability experiments included determination of gut fullness and feeding incidence of zebrafish Brachydanio rerio larvae fed on the two particle types. Acceptability of GAB by first-feeding zebrafish larvae was significantly greater than that for CLPWC. Acceptability of CLPWC increased in larger larvae and was similar to that for GAB. Both particle types were broken down by first-feeding zebrafish larvae. Growth experiments with CLPWC showed that up to 40% substitution of Artemia nauplii could be accomplished without reduced growth and survival of zebrafish larvae after a feeding period of 8 days. Twenty per cent substitution of Artemia nauplii could be achieved with GAB without reduced growth and survival.     

jnk3基因在金鱼和斑马鱼不同发育时期胚胎与成体不同组织中的分化表达

JNKs是丝裂原活化蛋白激酶(MAPK)家族重要成员,参与应激反应和动物体轴的构建。为了进一步了解JNK家族在动物发育中的功能,首次分离和克隆了金鱼jnk3基因,研究了jnk3在金鱼和斑马鱼组织特异性和胚胎发育特异性表达状况。金鱼jnk3基因cDNA总长1 794 bp,其中阅读框1 293 bp,编码430个氨基酸。对jnk3基因序列及其推导的氨基酸序列进行同源性分析,结果显示,jnk3基因在脊椎动物较为保守,金鱼jnk3基因核苷酸序列与斑马鱼、人的同源性分别为94.1%和80.7%,金鱼jnk3基因氨基酸序列与斑马鱼、人的同源性分别为99.7%和93.4%。jnk3基因在鱼类成体中的表达存在着显著的组织差异,在金鱼和斑马鱼脑和精巢组织的表达具专一性,此外在斑马鱼心脏中也检测到jnk3的表达。RT-PCR检测结果显示,在鱼类胚胎发育中,神经胚期开始检测到jnk3基因mRNA表达,从神经胚到心跳期胚胎,jnk3基因表达水平逐渐增高,此后直至出膜一直保持较高的表达水平。研究表明,jnk3基因可能在鱼类后期的胚胎发育和成体脑、精巢与心脏发育中具有重要作用。     

Effects of the prebiotic mannan‐oligosaccharide on feed deprived zebrafish: Growth and reproduction

This study was designed to investigate whether the prebiotic mannan‐oligosaccharides (MOS) can reduce the deleterious impacts of feed deprivation on growth and reproduction in zebrafish. In the growth performance experiment, juvenile fish were distributed in the following four treatments: normal‐control (NC), starved‐control (SC), normal‐prebiotic (NP) and starved‐prebiotic (SP). After 8 weeks, NP and SC fish showed the highest and lowest growth patterns, respectively among treatments. Standard length, specific growth rate and feed conversion ratio did not differ significantly between NC and SP treatments. Autochthonous lactic acid bacteria (LAB) were significantly higher in NP and SP than the NC and SP treatments. Feed restriction resulted in significantly lower concentrations of thyroxine. In the reproductive performance experiment, additional juveniles were fed as in the growth experiment. Crossing in the final week of the experiment resulted in successful spawning in only NP fish, which showed mature sperm and oocytes in histological examinations. The number of spermatozoa was significantly lower in the fish that experienced feed restriction; however, the oocytes of SP females were at the same maturation level to that observed in NC females. Sex‐steroids changed after both starvation and MOS supplementation where NC and SP showed no differences in the levels of testosterone and female's 17β‐estradiol. These results indicate that MOS supplemented diet reduced some side effects of feed deprivation (final weight and length, SGR, FCR and levels of LAB and T3), and suggest that supplementation of the diet with MOS may ameliorate some of the negative effects of feed deprivation in zebrafish.     

Cu（2＋）、Cd（2＋）、Cr（6＋）胁迫对斑马鱼抗氧化酶活性的影响

[目的]研究Cu2＋、Cd2＋、Cr6＋对斑马鱼抗氧化酶活性影响,探讨重金属对斑马鱼毒性大小。[方法]以斑马鱼为模式生物,采用半静态法对斑马鱼进行驯养和试验,测定Cu2＋、Cd2＋和Cr6＋对超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性影响。[结果]Cu2＋暴露1d胁迫斑马鱼SOD和CAT为诱导效应,胁迫POD为抑制-诱导-抑制效应,Cu2＋暴露7 d胁迫斑马鱼SOD为诱导效应,胁迫POD和CAT为抑制效应;Cd2＋和Cr6＋暴露1 d和7 d胁迫斑马鱼SOD和CAT均为诱导效应,胁迫POD为抑制效应。且暴露1 d和暴露7 d酶活性变化均显著。[结论]Cu2＋、Cd2＋和Cr6＋单一胁迫斑马鱼抗氧化系统酶活性变化显著,在短时间和低浓度暴露下,Cr6＋毒性作用最显著;在短时间中、高浓度暴露下,Cd2＋毒性作用最显著;长时间暴露时,Cu2＋毒性作用最强,可为重金属污染对水生生物毒性机理研究提供参考。     

斑马鱼血管紧张素转换酶(ACE)的生物信息学分析

采用生物信息学方法对模式生物斑马鱼中的血管紧张素转换酶进行了系列预测分析.结果表明,斑马鱼中有两个ACE基因,它们所编码的蛋白分别由1 324和785个氨基酸组成,其中1个是酸性蛋白,在水中不稳定,没有信号肽,属线粒体跨膜蛋白;另一个为酸性水溶性稳定蛋白,有信号肽.它们均含保守的Pfam Peptidase_M2结构域,属于M2家族蛋白.通过ESTs搜索发现这两个基因在斑马鱼不同组织和不同发育期的表达情况存在较大的差异.同源性比对和系统发育分析表明,斑马鱼ACE蛋白与低等脊椎动物(红原鸡、热带爪蟾等)、高等脊椎动物(人、鼠等)的同源性较高,达60%以上,说明ACE蛋白在系统演化上比较保守.