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斑马鱼雄核发育技术研究 总被引：1，自引：0，他引：1
为探索人工诱导斑马鱼雄核发育二倍体克隆鱼的途径和方法,运用紫外线照射卵子使其遗传物质失活,利用斑马鱼体表花纹形态作为遗传标记,使用热休克技术阻止受精卵的第一次卵裂加倍染色体。试验结果表明总辐射剂量为120mJ cm2紫外线照射斑马鱼卵可以使雌核遗传物质完全失活;体表花纹为豹斑花纹被证实是由纯合隐性基因控制的性状,可以作为雄核发育遗传标记;遗传失活卵受精后经2min,(41 5±0 5)℃的热休克后出现两个染色体加倍的高峰期,分别为受精后的11min和23min,且后一时期效果优于前一时期,最高二倍体诱导率达46 15%;经遗传分析,共获得正常存活的雄核发育二倍体克隆鱼19尾,说明经过优化的紫外线结合热休克等染色体操作技术,可以成为诱导雄核发育二倍体克隆鱼的有效手段。 相似文献
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草鱼呼肠孤病毒（GCRV）引起的草鱼出血病是造成我国草鱼养殖风险的主要疾病之一。II型草鱼呼肠孤病毒（GCRV-Ⅱ）是草鱼出血病的主要致病毒株,为了研究其流行规律,本文基于稀有鮈鲫感染GCRV-JX02的研究模型,利用分子生物学检测,RT-qPCR定量等方法开展GCRV-II水平和垂直传播的研究,结果显示,水平传播中浸泡感染和共培养感染都能使稀有鮈鲫成为GCRV-II的无症状携带者,阳性检出率分别为50%和80%;其中共培养感染直接导致8%的稀有鮈鲫死亡。感染后无症状的稀有鮈鲫经热休克处理,浸泡感染与共培养感染组的死亡率分别为57.14%和100%,总体阳性检出率分别为80.95%和100%。腹腔注射感染GCRV-JX02后存活的无症状稀有鮈鲫每0.01克精巢与卵巢中病毒拷贝数分别为3.64×106和6.84×106。垂直传播实验中稀有鮈鲫单个的卵子、未受精卵、受精卵和幼鱼的平均病毒拷贝数为1.98×103、1.15×104、4.75×103和6.74×104,幼鱼中病毒的拷贝数显著高于卵子与受精卵时期,表明GCRV-JX02不仅能够在稀有鮈鲫中垂直传播,还能伴随幼鱼的发育不断扩增。本研究阐明了不同传播途径下GCRV-Ⅱ的传播潜力,有助于评估草鱼出血病的流行风险,且为筛选阻断草鱼呼肠孤病毒传播的药物提供了有效的动物模型。 相似文献
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冷休克诱导大鳞副泥鳅雄核发育单倍体 总被引：4，自引：0，他引：4
为探索诱导鱼类雄核发育单倍体的新方法及形成机制,以大鳞副泥鳅为研究对象,无须卵子失活,仅用冷休克诱导雄核发育单倍体,并对其后代的早期胚胎发育、染色体数目、分裂胚细胞学观察等进行了系统分析。早期胚胎发育结果显示,0℃与3℃处理组的孵化率分别为0.83%和57.42%,二者之间孵化率差异显著(P0.05),0℃和3℃处理组均表现出单倍体综合征。染色体倍性鉴定结果显示,0℃处理组染色体分布为18~27,单倍体率为81.40%,3℃处理组染色体分布为20~30,单倍体率为68.97%。分裂胚细胞学观察结果显示,3℃处理组受精后60 min,卵核和第二极体位于胚表面,有一同释放迹象。结果表明,对大鳞副泥鳅受精卵进行3℃冷休克处理60 min,可以成功诱导雄核发育单倍体,研究推测冷休克诱导大鳞副泥鳅雄核发育的细胞学机制是雌核和第二极体一同释放。 相似文献
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热休克因子1(Heat shock factor 1,HSF1)作为转录调控元件,能够调控热休克蛋白(heat shock proteins,HSPs)的表达,保护机体免受压力损伤。为评估稀有鮈鲫(Gobiocypris rarus)HSF1和环境污染物五氯酚(pentachlorophenol,PCP)的关系,本研究采用cDNA末端快速扩增(RACE)技术从稀有鮈鲫肝脏中克隆HSF1全长序列,并将其命名为GrHSF1;利用Real-time PCR检测稀有鮈鲫GrHSF1的组织分布;温度以及PCP暴露下稀有鮈鲫肝脏中的GrHSF1的表达情况。RACE结果显示:GrHSF1的开放阅读框(open reading frame,ORF)长度为1581 bp,共编码527个氨基酸。系统发育树分析表明,预测得到的GrHSF1氨基酸序列与其他鱼类HSF1具有广泛的序列相似性。组织分布显示GrHSF1在稀有鮈鲫的12种组织中均表达,在肝脏中表达量最高。在肝脏中的暴露实验表明,当80μg/L≥PCP≥8μg/L时,GrHSF1的mRNA与PCP之间具有明显的剂量依赖效应,当160μg/L≥PCP≥8μg/L时,GrHSF1的mRNA与PCP之间具有明显的时间依赖效应;在过高或过低温度下,GrHSF1的mRNA均显著下降。结果表明GrHSF1是已知鱼类HSF1家族的同源基因,并可作为评估环境污染物PCP的生物标志物。 相似文献
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采用紫外线照射灭活精子,经冷休克诱导染色体加倍的方法对翘嘴鳜(Siniperca chuatsi)进行人工诱导雌核发育育种。灭活翘嘴鳜精子,紫外线照射剂量80 mJ/cm2是最佳的辐射剂量。冷休克结果表明,在0～3℃冰水下,冷休克起始时间为授精后3～7 min均有效,以5 min为最适;冷休克持续时间10-50 min均有效,以30 min最佳。综合以上两项因素,授精5 min后,休克时间持续30 min组的原肠胚存活率和孵化率均为最高,与其他处理组差异显著(P<0.05),此时雌核发育率最高,达到4.72%。 相似文献
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施氏鲟精子诱导匙吻鲟雌核发育 总被引：4，自引：1，他引：3
采用照射强度为10 188 μW/(cm2·s),照射距离为15 cm的紫外线对施氏鲟(Acipenser schrenckii)精子进行4～7 min的照射,用照射后的精子对匙吻鲟(Polyodon spathula)卵进行人工授精,以探索精子的最佳紫外线照射时间.设计热休克起始时间、持续时间和休克温度3因子、3水平的正交试验,以探索人工诱导匙吻鲟雌核发育的理想热休克条件.结果表明:用紫外线照射5 min处理的施氏鲟精子,与匙吻鲟卵受精所得胚胎经染色体观察均为单倍体(n=60),表明照射精子遗传失活的有效性和可靠性;若热休克处理前受精卵保持在(18±013)℃,在一定的休克温度(35～39℃)条件下,于不同时间(受精后16～20 min)起始热休克,持续处理2 min均能诱导受精卵染色体加倍.对热休克处理条件的优化结果表明,将受精卵于受精后18 min,在37℃的水中热休克处理2 min,二倍体诱导率最高,达18.8%.染色体鉴定显示,该条件下处理的胚胎均为二倍体(2n=120),未发现单倍体和非整倍体.本研究为进一步分析异源精子诱导匙吻鲟雌核发育机制以及匙吻鲟性别决定的分子机制奠定了基础. 相似文献
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鱼类个性行为反映其生活史策略,研究环境因素对鱼类个性行为的影响,可为更好地理解该行为的适应性意义及优化鱼类养殖环境提供理论依据和数据基础。以稀有鮈鲫（Gobiocypris rarus）为研究对象,使用隔板跨越和遮蔽物出游2 种实验方法,观测了同类鱼群刺激、禁食及不同环境照度下稀有鮈鲫的个性行为。结果显示,隔板跨越测试中,稀有鮈鲫开始运动前的潜伏期在同类鱼群存在与否的情况下没有显著差异（P>0.05）,但在禁食后显著减小（P<0.05）,在不同照度下差异显著（P<0.05）,且随照度的增加而增大;稀有鮈鲫总跨越隔板数在不同的照度条件下差异极显著（P<0.01）,随照度的增加而减小,但在同类鱼群存在与否及禁食组与饱食组之间差异均不显著（P>0.05）。遮蔽物出游测试中,稀有鮈鲫游出遮蔽物的时间在24 h 禁食后变化不显著（P>0.05）,但在同类鱼群出现时差异显著（P<0.05）,不同照度条件下差异极显著（P<0.01）,且随照度的增加先减小再增大。研究表明,禁食导致的饥饿和同类鱼群的出现能够在一定程度上增强稀有鮈鲫的探索性,但环境照度是稀有鮈鲫个性行为最为关键的影响因子。 相似文献
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真鲷精子诱导牙鲆减数分裂雌核发育 总被引：12，自引：0，他引：12
采用真鲷精子激活牙鲆卵子,经(0±0.5)℃冷休克处理,诱导了牙鲆减数分裂雌核发育。灭活真鲷精子,紫外线照射剂量3.4mJ/cm2时孵化率最低;随着照射剂量的增加,孵化率恢复,照射剂量73mJ/cm2时,孵化率最高,呈现典型的Hertwig效应。用流式细胞仪检测倍性,未经冷休克处理的胚胎全部为单倍体,经冷休克处理的均为二倍体,表明精子灭活有效。冷休克实验结果表明,冷休克起始时间2～5min均有效,以3min为最好;冷休克持续时间30～90min均有效,以45min为最好。综合两项因素,授精3min后,休克时间持续45min组的授精率和孵化率均为最高,与其他处理组差异显著(P<0.05)。RAPD分析结果显示,异源精子的遗传信息未在雌核发育二倍体的电泳图中出现,表明其遗传信息没有传递给子代。 相似文献
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异源精子诱导犬齿牙鲆的雌核发育 总被引：1，自引：1，他引：0
用冷冻保存的花鲈精子作为异源精子,采用紫外线(UV)对精子进行照射使其遗传物质失活,然后与卵子进行"授精",可以刺激犬齿牙鲆鱼卵进行雌核发育,在受精后一定时间采用冷休克处理"授精"卵,抑制第二极体排出成功获得了犬齿牙鲆雌核发育二倍体鱼苗.实验表明:犬齿牙鲆同源精子经80 mJ/cm2的紫外线照射可以完全失活,冻存花鲈精子经紫外线照射后也同样具有诱导犬齿牙鲆雌核发育的能力.无论同源精子还是异源精子,最佳诱导条件为在18℃条件下,精子经80 mJ/cm2的紫外线照射,受精后4～5 min将受精卵放在3℃海水中进行冷休克处理,持续时间为45 min,同源精子和异源精子二倍体诱导分别为32.66%±7.03%和28.00%±6.48%.采用形态学、流式细胞仪DNA含量分析和微卫星标记技术对雌核发育鱼苗进行了分析,证明了雌核发育鱼苗为雌核发育二倍体. 相似文献
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Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean 
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下载免费PDF全文 The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna. 相似文献
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Liping Li Rui Wang Wanwen Liang Xi Gan Ting Huang Yan Huang Jian Li Yunliang Shi Honglin Luo Ming Chen 《广西水产科技》2014,(1)
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. 相似文献
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In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg. 相似文献
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Growth hormone (GH) and reproduction: a review 总被引：5，自引：0，他引：5
Florence Le Gac Odile Blaise Alex Fostier Pierre-Yves Le Bail Maurice Loir Brigitte Mourot Claudine Weil 《Fish physiology and biochemistry》1993,11(1-6):219-232
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. 相似文献
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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. 相似文献
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Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection 下载免费PDF全文
下载免费PDF全文 G Sharon M Pimenta‐Leibowitz M C L Vilchis N Isakov D Zilberg 《Journal of fish diseases》2015,38(1):67-74
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). 相似文献
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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. 相似文献
19.
Sandra García‐Garrido Ismael Hachero‐Cruzado Carlos Rosas Pedro Domingues 《Aquaculture Research》2013,44(11):1741-1751
Protein and amino acid composition of the mantle of juvenile O ctopus vulgaris (Cuvier, 1797) during fasting for 27 days were determined. Average protein content of octopus mantle was of 711.19 ± 46.80 g kg?1 DW, and it decreased with increasing fasting days. The non‐essential amino acids content was higher (486.18 ± 11.08 g kg?1 protein) than essential amino acids (425.82 ± 9.15 g kg?1 protein) at the start of the experiment (unstarved animals). The results suggest that the amino acid profile of the mantle where the most abundant amino acids are Arg, His, Lys, Gly, Leu and Pro could indicate a prolonged fasting condition (>20 days) or poor nutrition of O . vulgaris. This study supports the idea of using mantle for metabolic needs of starved O . vulgaris suggesting that the degradation pathway of amino acids to pyruvate and tricarboxylic acid cycle intermediates was favoured contrary to the degradation pathway of ketogenic amino acids. Special considerations should be taken concerning Thr, Ile, Ser, Ala, Asx (Asp, Asn), Glx (Glu, Gln) (because of their fast intake) and Lys and His (due to their stable contents) during a prolonged period of fasting. 相似文献
20.
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. 相似文献