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1.

利用cDNA末端快速克隆方法获得了青虾(Macrobrachium nipponense)的过氧化物还原酶基因(Prx)全长cDNA序列。该基因cDNA全长878 bp, 包括72 bp5′末端非翻译区, 594 bp的开放阅读框(ORF), 212 bp3′末端非翻译区, 开放阅读框编码198个氨基酸。氨基酸相似度比对显示, 所分离的青虾过氧化物还原酶基因包括两个半胱氨酸残基的区域FYPLDFTFVCPTEI”GEVCPA”。系统进化树分析表明, 青虾过氧化物还原酶基因与南美白对虾(Litopenaeus vannamei)过氧化物还原酶聚在一起, 具有最近的亲缘关系。荧光定量PCR检测显示, 过氧化物还原酶基因在青虾不同组织中均有表达, 其表达量由低到高依次为肠道、心脏、卵巢、肌肉、鳃、肝胰腺。使用荧光定量PCR检测青虾在低氧胁迫和复氧条件下肝胰腺中的过氧化物还原酶基因mRNA的时空表达情况, 结果显示, 与对照组相比, 实验组青虾过氧化物还原酶在肝胰腺和鳃组织中的表达量分别在低氧胁迫12~24 h 和复氧6 h出现了3次明显上调, 由此推测过氧化物还原酶基因参与低氧应激分子过程。本研究结果可为进一步了解青虾低氧应激分子机制提供参考。

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2.
嗜水气单胞菌灭活疫苗免疫后大鲵外周血免疫指标的变化   总被引:1,自引:1,他引:0  

以福尔马林灭活的嗜水气单胞菌(Aeromonas hydrophila)作为免疫原(F-AH), 通过腹腔注射免疫健康大鲵(Andrias davidianus), 分别于免疫后的第147142128天采集血液样品, 进行大鲵外周血液的血细胞计数与白细胞组成分析, 测定吞噬细胞的吞噬活性以及血清中和抗体效价, 于免疫28 d后进行攻毒感染试验。结果表明, 与对照组相比, 在免疫后第4714和第21, 免疫大鲵外周血中血细胞数量显著增加, 其中, 红细胞和白细胞数量均于第4天达峰值, 分别为7.83×107/mL6.74×106/mL; 嗜中性粒细胞百分比也在第4天达峰值, 28.60%, 单核细胞百分比在第7天达峰值, 10.53%; 吞噬细胞活性显著提高, 且吞噬百分比和吞噬指数均在第4天达到最高值, 分别为34.09%3.73。随后, 淋巴细胞百分比和中和抗体效价显著增加, 均在第21天达峰值, 分别为75.30%1426.67。攻毒感染实验结果表明, 免疫组的相对免疫保护率为69.23%。由此可见, F-AH免疫原能够通过促进血细胞数量的增加、吞噬细胞吞噬活性增强以及特异性抗体的产生等方式提高大鲵的免疫保护力。

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3.

应用实验生态学的方法研究斜带石斑鱼(Epinephelus coioides)(♀)×鞍带石斑鱼(E. lanceolatus)(♂)杂交子代(青龙斑)仔、稚鱼阶段的生长模式。利用Q-Capture Pro 6软件对仔、稚鱼(0~28日龄)的全长、肛前长、体高、头长、眼径、口径、第二背鳍棘、腹鳍棘、胸鳍、臀鳍和尾鳍等11个可量性状进行拍照和测量。研究表明青龙斑全长的生长可分为3个阶段, 不同阶段生长速率存在显著差异(P<0.05)。各功能器官均表现为异速生长。在11个可量性状中, 肛前长为等速生长, 头长和体高的生长由正异速生长分别转变为等速生长和负异速生长; 头部器官(口径和眼径)生长在20 ~22日龄时出现拐点, 拐点后分别转变为等速生长和负异速生长; 运动器官在拐点前均为正异速生长, 除臀鳍外, 其他各鳍的生长均存在不同的拐点。通过对青龙斑仔、稚鱼异速生长的研究, 发现在早期发育过程中, 有关摄食、感觉、运动等功能器官得到优先发育。在人工繁殖苗种的培育中, 可根据青龙斑重要器官发育的优先性, 创造有利的外部环境, 提高苗种的成活率。

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4.

对水产品中呈味核苷酸和氨基酸的高效液相色谱检测方法进行了优化, 并对烟台近海长牡蛎(Ostrea gigas)、栉孔扇贝(Chlamys farreri)和中国蛤蜊(Mactra chinensis)3种贝类中呈味核苷酸和氨基酸的含量进行了比较分析, 从而为开发贝类复合调味料提供科学依据。以纯水提取呈味物质, 采用DABS-Cl柱前衍生反相高效液相色谱法测定游离氨基酸, 流动相为17 mmoL/L柠檬酸溶液(pH 6.4, 4%DMF)和乙腈溶液(4%DMF), 梯度洗脱, 检测波长436 nm反相离子对色谱法测定呈味核苷酸, pH 4.5的磷酸二氢钾溶液和乙腈溶液为流动相, 磷酸二氢钾溶液内含5 mmoL/L四甲基氢氧化氨(TMAOH), 梯度洗脱, 检测波长254 nm。方法加标回收率为90.2%~108%, 相对标准偏差(RSD)0.7%~7.1%。准确度、精密度均满足分析要求。测定结果显示: 长牡蛎中牛磺酸、丙氨酸和谷氨酸含量较高, 栉孔扇贝中甘氨酸、牛磺酸和丙氨酸含量较高, 中国蛤蜊含有较多的甘氨酸、牛磺酸和丙氨酸; 3种贝类中, 丙氨酸、谷氨酸的TAV均大于1, 栉孔扇贝和中国蛤蜊中, 甘氨酸的TAV大于1, 对呈味具有显著贡献。呈味核苷酸中, 长牡蛎中IMP含量最高, 占总呈味核苷酸的80%, 其次是GMP。中国蛤蜊中AMP含量最高, 其次是IMP。而栉孔扇贝中仅检测到IMP3种贝类中呈味核苷酸含量虽然都较低, TAV未超过1, 但核苷酸之间、核苷酸和氨基酸之间的协同增效作用, 会对贝类的呈味产生重要影响。本研究通过对烟台近海3种贝类中各类鲜味物质和多种辅助呈味物质的含量、比例关系及其相互作用规律进行探讨, 旨在为充分利用贝类营养氨基本作为复合调味料及其功能性开发和评价提供科学依据。

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5.
斑鳜精液超低温冷冻保存及其效果分析   总被引:1,自引:1,他引:0  

实验所用翘嘴鳜(♀, Siniperca chuatsi) 2~3, 体质量1 000~1 500 g; 斑鳜(♂, Siniperca scherzeri Steindachner)1~2, 体质量300~500 g。于繁殖季节选取成熟度好的亲鱼以促黄体素释放激素类似物(LHRH-A2)、绒毛膜促性腺激素(HCG)和马来酸地欧酮(DOM)进行催产。通过筛选D-15D-17Ringer M-Hank’s4种稀释液和二甲基亚砜 (DMSO) 甘油(Gly)、乙二醇 (EG)1,2-丙二醇 (PG)和二甲基甲酰胺 (DMF)5种抗冻剂, 发现DMSOD-17分别是优选的抗冻剂和稀释液。以D-17作为斑鳜精子的稀释液, 按照13(精液稀释液)体积比稀释, 添加10%体积的DMSO作为抗冻剂, 按照三步冷冻法超低温冷冻保存, 37水浴解冻的斑鳜精子, CASA分析精子活率为(83.26±18.20)%, SCGE检测表明70%以上的精子核DNA不会发生损伤; FCM分析表明26.74%的解冻精子有完整的细胞膜且线粒体功能正常; 用冷冻复苏斑鳜精子与翘嘴鳜精卵进行人工授精, 最高受精率(39.6±6.5)%, 温度24~28孵化时间38 h, 孵化后的鱼苗发育正常, 开口1周以后的鱼苗全长(1.346±0.255 ) cm, 体高(0.438±0.103) cm, 体质量(0.045±0.020) g。鲜精受精鱼苗和冻精受精鱼苗在开口后1周的生长没有显著差异。因此认为D-17+10% DMSO可用于斑鳜精液的超低温冷冻保存。本研究将有助于斑鳜种质资源的收集保存和冷冻保存的斑鳜精液在翘嘴鳜(♀)×斑鳜(♂)杂交中的应用。

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6.

以鲢(Hypophthalmichthys molitrix)肌肉cDNA为模板, 利用小清蛋白特异性引物进行PCR扩增, 克隆得到β小清蛋白两种不同亚型, 型、型编码区基因。将目的基因片段连接到pET28a (+)表达载体, 并在大肠杆菌[E.coli BL21 (DE3)]中诱导表达。结果表明, 经诱导的小清蛋白重组质粒菌株有特异的蛋白表达。SDS-PAGE分析显示, 目的蛋白的分子量约为13 kD, 与预期大小一致。菌体超声破碎后发现2种亚型的小清蛋白均为可溶表达。利用Ni2+亲和层析柱对重组蛋白进行纯化, 得到高纯度的重组小清蛋白PVⅠPVⅡ。经Western Blot 鉴定, 重组小清蛋白PVⅠPVⅡ均能与抗鲢小清蛋白单克隆抗体反应。本研究为进一步分析小清蛋白的结构与致敏性的关系提供了重要的基础。

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7.

选用750尾初体质量为(114.2 ± 6.5) g的大菱鲆 (Scophthalmus maximus L.), 随机分成5, 每组3个重复, 每重复50尾鱼, 分别投喂在高脂基础饲料中添加0(D-0)120 (D-120)240(D-240)480 (D-480)960 mg/kg (D-960) DL-α-生育酚醋酸酯5种试验饲料养殖110 d, 考察维生素E(VE)对高脂饲料养殖大菱鲆生长、脂类代谢和抗氧化性能的影响。结果表明, D-240组大菱鲆末均质量最高, 且显著高于D-0(对照组)(P<0.05), 与其他组差异不显著(P>0.05); D-120组和D-240组饲料系数显著低于D-0(P<0.05), 但蛋白质效率显著高于D-0(P<0.05)D-0组和D-960组血清总胆固醇、甘油三酯、低密度脂蛋白胆固醇和谷丙转氨酶显著高于其他各组(P<0.05), D-0组和D-960组高密度脂蛋白胆固醇则显著低于其他各组(P<0.05); 随着饲料VE添加水平的增加, 大菱鲆肝脂蛋白脂酶(LPL)活性有降低的趋势; 脂肪酸合成酶(FAS)活性, 有先增加后降低的趋势。大菱鲆肝总抗氧化酶、超级氧化歧化酶和过氧化氢酶活性, 均随VE添加水平的升高有先升高后下降的趋势, 而丙二醛含量的变化趋势则相反; 随着饲料VE水平的增加, 大菱鲆肝VE积累量有显著升高的趋势(P<0.05), 肌肉VE的积累量先升高后恒定在一个水平。综上所述, 在高脂饲料中添加适量的维生素E (有效含量124~243 mg/kg) 能改善大菱鲆生长性能、饲料利用效率、鱼肝体指数、脏体指数, 调节和改善血脂代谢, 可调控机体脂肪代谢酶活性, 提高机体组织VE积累量和肝抗氧化功能。本研究旨在通过考察添加不同水平的VE对高脂胁迫下大菱鲆生长、脂类代谢酶、抗氧化能力的影响, VE在高脂胁迫下的合理使用及以VE营养调控鱼类脂肪代谢和改善抗氧化功能提供参考依据。

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8.

开展银鲳(Pampus argenteus)试养海域的盐度易随气候降低, 且受到一定程度Cu等重金属污染, 研究低盐条件下硫酸铜对银鲳的影响十分必要。本实验首先进行银鲳幼鱼低盐度适应, 将盐度以4的幅度从24逐步降低至12。稳定后进行硫酸铜胁迫, 在盐度12CuSO4·5H2O浓度梯度设为00.10.30.5 mg·L–1, 盐度24CuSO4·5H2O浓度梯度设为00.5 mg·L–1, 胁迫持续144 h。通过检测两种鳃离子调节酶: Na+/K+-ATP(NKA)V-H+-ATP(VHA), 以及3种肝抗氧化活性物质-还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT), 探究低盐条件下铜离子对银鲳幼鱼上述指标的影响。结果显示, 盐度逐步降低后, NKAVHA活力呈上升后下降的变, 之后NKA活力随硫酸铜浓度增加而减弱; VHA活力加入硫酸铜后都出现显著下降, 其中CuSO4·5H2O 0.3 mg·L–10.5 mg·L–1组在72 h时下降更为明显; 盐度24硫酸铜组NKAVHA活力都在24 h时增强而后减弱。GSH含量和SOD活力在盐度降低时出现跃升, CAT活力则呈波动变化, 加入硫酸铜后, 0.3 mg·L–10.5 mg·L–1GSH含量持续下降后显著上升而后回落; 各硫酸铜组SOD活力都出现增强后回落的变化; 0.3 mg·L–10.5 mg·L–1CAT活力在72 h有显著增强。本实验表明, 铜离子对NKAVHA有抑制作用, 盐度降低时其作用更强; GSHSODCAT的变化能反映低盐度和铜离子对银鲳的伤害程度。银鲳对水体铜离子有一定的抗性, 但在盐度降低等环境变化时, 应当密切注意水体中铜等重金属浓度。

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9.

本研究采用cDNA末端快速扩增法(RACE)克隆了草鱼(Ctenopharyngodon idellus)IGF-IR基因全长cDNA序列, 并对该基因在草鱼不同时期胚胎和成鱼不同组织中的表达进行了分析。序列分析表明, 草鱼IGF-IR基因cDNA序列全长5 741 bp, 包括5′端非翻译区822 bp, 3′端非翻译区581 bp, 开放阅读框4 338 bp, 共编码1 445个氨基酸。序列比对结果显示, 草鱼IGF-IR可能属于a, 该基因编码的氨基酸序列与鲤(Cyprinus carpio)IGF-IRa、斑马鱼(Danio rerio)IGF-IRa和人类(Homo sapiens)IGF-IR的相似性分别为95%93%66%, 具有较高的同源性, 表明该基因在长期进化中具有较高的保守性。RT-PCR结果表明, 该基因从16 hpf(hours post fertilization)胚胎期到出苗期都有表达, 在成鱼大部分组织中均有表达。原位杂交结果显示, 草鱼IGF-IR mRNA在不同时期胚胎组织中广泛存在, 其中在脑部、脊索和尾部等生长旺盛组织的细胞中表达量较高。本研究为进一步探索草鱼IGF-IR基因在生长发育信号通路中的作用和育种提供了基础资料。

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10.
大菱鲆选育家系抗鳗弧菌性能   总被引:5,自引:2,他引:3  

基于家系选育技术开展了大菱鲆(Scophthalmus maximus)抗鳗弧菌病选育研究, 2010年构建的37个选育二代家系中选择成活率高的30个家系进行鳗弧菌(Vibrio anguillarum)感染实验, 开展对鳗弧菌的抗病性研究。结果表明, 不同家系间抗鳗弧菌感染能力存在着显著差异(P0.05); 30个家系中, 鳗弧菌感染后, 12个家系的存活率达到65% 以上, 其余家系的存活率则低于65%; 通过bCOX回归分析, 计算各家系的优势比, 优势比最高的5个家系的存活率达到65% 以上; 各家系的成活率、优势比和死亡历时差4项指标进行聚类分析, 优势比最高且存活率达到65% 以上的5个家系聚为一类。综合感染家系的高成活率、高优势比以及聚类分析的结果, 选育出5个抗病力较强的优良家系。选育出的抗病力较强的家系可做为抗鳗弧菌选育的核心育种群体, 为抗鳗弧菌病的传代选育奠定良好的基础。

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Anguilla luzonensis and A. huangi were each described in 2009 using eels obtained from northern Luzon Island. We examined the taxonomic status of these two groups of eels using morphological and molecular genetic characters. There were no significant differences in two vertebrae counts between eels of A. luzonensis and A. huangi. Mitochondrial 16S ribosomal RNA and cytochrome b genes sequences were obtained and compared among 28 specimens of A. luzonensis, the holotypes of A. luzonensis and A. huangi, and one specimen of the other 15 anguillid species. The specimens of A. luzonensis exhibited almost identical sequences, including the holotype, with only a few site differences, and the genetic difference between the holotypes of A. luzonensis and A. huangi was within the range of differences of specimens of A. luzonensis. The other anguillid species were genetically very different from A. luzonensis and A. huangi, although A. interioris is a closely related species. It is clear that A. luzonensis and A. huangi are the same species, and according to the principle of priority in zoological nomenclature, A. luzonensis Watanabe, Aoyama, and Tsukamoto, 2009 is the valid species name, and A. huangi Teng, Lin, and Tzeng, 2009 is a junior synonym of A. luzonensis.  相似文献   

13.
The effects of exposing the eggs of Pacific threadfin and amberjack eggs (AEs) to different concentrations of hydrogen peroxide for 5 min on hatch rate and survival were assessed in a series of experiments using a petri dish model rearing system. Despite significant inter‐batch variation in hatch rate, it was shown that eggs of both species could be safely exposed to up to 11 340 mg L−1 H2O2 for 5 min. Exposure to 34 230 mg L−1 H2O2 for 5 min was shown to be lethal to AEs at a late stage of development. In two further experiments, it was demonstrated that Pacific threadfin eggs were resistant to all tested concentrations of a range of polyvinylpyrrolidone iodine (PVP‐I) concentrations and contact times (up to 1000 mg L−1 PVP‐I for 10 min). The level of bacteria adhering to the eggs of both species was highly variable. Where eggs were heavily colonized (>104 cfu egg−1), hydrogen peroxide concentrations of at least 11 340 mg L−1, or PVP‐I concentrations higher than 500 mg L−1 for 10 min, were required for effective sterilization. In less colonized batches, rinsing in sterile seawater or exposure to lower (550 mg L−1) concentrations of H2O2 was sufficient to result in high apparent levels of surface sterility (<1 cfu egg−1).  相似文献   

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Mulloway (Argyrosomus japonicus) occur in estuarine and coastal waters surrounding Australia, Africa, India, Pakistan, China, Korea and Japan, where they are important in fisheries. This study identified that mulloway in south-eastern Australia had similar growth rates, but matured at smaller lengths and younger ages, to those in South Africa and Western Australia. Growth of both sexes was similar to about 5 years, after which females grew faster and attained a greater maximum length than males. Female mulloway matured at 4–5 years of age with a L50 of 68 cm, whereas males matured at 2–3 years of age with a L50 of 51 cm. The commercial fishery in New South Wales was characterised by declining catches and a reduction in the proportion of mulloway of mature lengths in landings. During 2002–2005 commercial landings were dominated (83%) by fish within 15 cm of the current minimum legal total length of 45 cm and aged 2 and 3 years (>80%), even though mulloway can attain lengths of 200 cm and live >30 years. Estimates of the rates of instantaneous total mortality ranged between 0.34 and 0.45, whilst the rate of instantaneous natural mortality (M) was estimated to be approximately 0.12. Yield-per-recruit analyses indicated that mulloway in New South Wales are being growth overfished and substantial increases in yield could be achieved by increasing the length at first harvest. Values of the spawning potential ratio were below 0.2 under a range of mortality estimates, suggesting that mulloway are at risk of recruitment overfishing. These results suggest that the spawning stock of mulloway in south-eastern Australia has been depleted and that remedial management action is required to protect this iconic species.  相似文献   

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In an investigation of the potential for pearl production in New South Wales (NSW), Australia, changes in the physical and reproductive condition of the pearl oyster, Pinctada imbricata, were monitored for over 2 years. Using wild oysters gathered from close to the southern extent of the species' range in Port Stephens, NSW, a series of macroscopic and histological observations were made. Reproductive activity in P. imbricata was greatest from late spring to early autumn with oysters in poor reproductive condition during winter. Peaks in reproductive indices occurred in November 1998, March 1999, December 1999 and April 2000. Four indices of physiological condition were used: shell growth, byssal attachment, mantle thickness and mucoprotien layer. With the exception of the thickness and extent of the mucoprotein layer, these indices either showed little variation or the variation that occurred was not related to seasonal or reproductive changes. Changes in the mucoprotein layer were correlated with water temperature and suggest that this layer is metabolized during periods of high demand such as during gonadogenesis. Collectors deployed at two sites in Port Stephens demonstrated that spatfall occurs largely in the months of December and January following the spring and early summer peaks in reproductive activity (November 1998 and December 1999). Spatfall was not observed following the autumnal peaks (March 1999 and April 2000) in reproductive activity. Overall, reproductive patterns in P. imbricata are poorly suited to spat supply in Port Stephens. Farmers require spat in early spring (September) to allow maximum use of the ‘growing’ season (September–May). Reproductively capable oysters are not available from the wild until September and quantity of natural spatfall is too variable and occurs too late in the season (December–January). As a result, oysters are being conditioned in the hatchery in July, spawned in August and spat are supplied to farmers in mid September.  相似文献   

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对驼背鲈(Chromileptes altivelis)的胚胎发育及仔、稚、幼鱼的形态特征进行了详细的观察与研究,描述了从受精卵到仔、稚、幼鱼各发育期的时间和形态特征变化。结果表明,在水温25-26℃、盐度30的海水中,受精卵历时27 h 25 min完成整个胚胎发育过程,经历从卵裂、囊胚、原肠、神经胚到肌节形成、各器官的逐渐形成、变化、完善等一系列的胚胎发育和变化过程;根据其卵黄囊消长情况、鳞片的覆盖状态、体色发生的不同变化以及第二背鳍和腹鳍的消长,将胚后发育分为仔、稚、幼鱼3个阶段。在水温22-26℃、盐度29-31、DO≥5 mg/L的条件下,2-3 d仔鱼卵黄囊消失,开口摄食;生长发育至31 d,仔鱼已变态进入稚鱼期;培育至57 d,稚鱼完成变态,成为幼鱼。第二背鳍棘和腹鳍棘的生长与收缩等石斑鱼类早期发育的共性生长特征及其体表特性体色变化特征为驼背鲈胚后发育过程中最明显的特征。  相似文献   

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Mosquitofish, Gambusia sp., have been spread throughout the world to biologically control mosquitoes. However, the fish has gained a reputation as an invasive species and has been implicated in displacing native aquatic species. Gambusia affinis are native to the southeastern United States and commonly occur in commercial channel catfish, Ictalurus punctatus, production ponds. We investigated effects of mosquitofish presence on zooplankton populations, water quality, disease occurrence, and fish production in experimental ponds. There were no differences between ponds with or without mosquitofish in numbers of calanoid copepods, cyclopoid copepods, total copepods, Bosmina sp., Ceriodaphnia sp., Moina sp., Daphnia sp., or total cladocerans. There were also no differences in copepod and cladoceran sizes. Copepod nauplii were more numerous during the summer months in ponds with mosquitofish. There were no differences in water quality variables (soluble reactive phosphorus, nitrate, nitrite, ammonia, total nitrogen, total phosphorus, pH) or phytoplankton density between ponds stocked with and without mosquitofish. Catfish production and disease occurrence were also similar between ponds with and without mosquitofish. Although mosquitofish may cause problems when stocked outside their native range, there does not appear to be any adverse effects of mosquitofish presence in catfish production ponds.  相似文献   

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