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镢鱼病毒核酸随机引物扩增与克隆 总被引：1，自引：0，他引：1
从感染鳜鱼病毒（Ｓｉｎｉｐｅｒｃａ ｃｈｕａｔｓｉ Ｖｉｒｕｓ,简称ＳＣＶ）的鳜鱼体中分离病毒,提纯核作模板。通过ＲＡＰＤ法,和带酶切位点的引物进行病毒核酸随机引物扩增,获得扩增带谱。从带ＥｃｏＰＩ酶切位点的引物扩增产物中,用琼脂糖凝胶电泳回收大小约为０．４和０．５Ｋｂｐ的片段,经ＥｃｏＲＩ酶切处理制备成带粘性未端的片段,分别插入质粒ｐＵＣ１９的ＥｃｏＰＩ位点。ＥｃｏＲＩ酶对重组子进行了酶切鉴定, 相似文献
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鳜鱼病毒核酸的初步分析 总被引：7，自引：1，他引：6
提取鳜鱼病毒核酸,分别用ＤＮａｓｅ、ＲＮａｓｅ和绿豆芽核酸酸处理表明,该病毒为双链ＤＮＡ病毒,用带ＥｃｏＲⅠ酶切位点的随机引物扩增病毒核酸,获得扩增核酸带谱。经低熔点琼脂糖回收ＰＣＲ产物,与质粒ｐＵＣ１９连接,获得三个重组子,已经对两个小插入片断进行了序列分析,插入序列分别为３６９ｂｐ和４５０ｂｐ。ＧｅｎＢａｎｋ检测显示,尚未有类似的序列报导。 相似文献
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从ＲＡＰＤ扩增的鳜鱼病毒（ＳＣＶ）核酸电泳带中回收了二个片断，克隆子pUC19质粒（称为ＳＣＶＥ３６９和ＳＣＶＥ４５０），序列分析表明插入片段分别为３６９bp和450bp与ＧenBank序列没有显著的同源，根据克隆序列调计两对引物Ｐ１／Ｐ２和Ｐ３／Ｐ４ ，在健康鳜鱼，病鳜以及提纯的ＳＣＶ核酸中进行ＰＣＲ试验，结果表明，Ｐ１／Ｐ２组引物在ＳＣＶ基因组中扩增出特异性核酸片段，可作为鳜鱼病毒ＰＣＲ诊断，检测片段为３６９bp. 相似文献
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对虾病毒HHNBV DNA构建质粒的研究与分析 总被引：2，自引：0，他引：2
用ＰＵＣ１８构建了对虾病毒ＨＨＮＢＶＤＮＡ重组质粒，提取后经斑点杂交及酶切分析，证实质粒中插入片段为病毒ＤＮＡ，其中０５＃质粒的插入片段大于２Ｋｂ，与质粒的分子量相近。对０５＃质粒酶切后的Ｓｏｕｔｈｅｒｎ杂交亦取得了满意的结果。同时，还对０５＃质粒的插入片段进行了内切酶图谱分析，共进行了ＥｃｏＲｌ、Ｈｉｎｄｌｌｌ、Ｓｍａｌｌ、Ｐｓｔｌ、ＰＵＶｌ、Ｈｉｎｄｌｌ６种限制性内切酶分析，结果表明，只有Ｐｓｔｌ在插入片段的一端有一个酶切位点，酶切位点的确切位置有待于进一步确定。 相似文献
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斑节对虾白斑综合症病毒部分基因组文库及核酸探针检测法 总被引：12，自引：1，他引：11
通过分离纯化白斑综合症病毒（ＷＳＳＶ）粒子,抽提病毒ＤＮＡ。用限制性内切酶ＥｃｏＲⅠ或ＳａｌⅠ酶切后,克隆入质粒ｐＢｌｕｅｓｃｒｉｐｔⅡＫＳ中,从而建立了ＷＳＳＶ部分基因组文库。估计ＷＳＳＶ基因组ＤＮＡ在１６５ｋｂ以上。将ＷＳＳＶ ＥｃｏＲⅠ克隆片段标记制备为探针。进行Ｓｏｕｔｈｅｒｎ杂交、打点杂交和原位杂交,其结果证明了克隆片段对ＷＳＳＶ特异,并为检测ＷＳＳＶ提供了方法。通过对部分基因组文库序列 相似文献
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用６种限制性内切酶分析了４条条纹斑竹鲨的线粒体ＤＮＡ，ＰｓｔＩ，ＨｐａＩ，ＸｂａＩ，ＥｃｏＲＩ，ＥｃｏＲＶ，ＢｇｌⅡ在条纹斑竹鲨ｍｔＤＮＡ分子上分别具有０至２个切点，ｍｔＤＮＡ分子大小为１６．６ｋｂ，根据单酶和双酶完全酶解片段的大小，构建了条纹斑竹浙江省ｍｔＤＮＡ的限制性酶切图谱。 相似文献
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用ＡｐａⅠ，Ｂｃ１Ⅰ，Ｂｇ１Ⅱ，ＤｒａⅠ，ＥｃｏＲⅠ，ＥｃｏＲＶ，ＨｉｎｄⅢ，ＫｐｎⅠ，ＰｓｔⅠ，ＰｖｕⅡ，ＳａｃⅠ，ＸｂａⅠ，ＸｈｏⅠ等１３种限制性核酸内切酶对来自尼罗河上游和下游的两种不同地方种群尼罗罗非鱼的线粒体ＤＮＡ（ｍｔＤＮＡ）进行酶切分析，得到这两种罗非鱼的ｍｔＤＮＡ分子大小为：上游尼罗为１６９０９±９０ｂｐ，下游尼罗为１６９１２±１００ｂｐ。研究中还发现我国养殖的这两种不同地方种群尼罗罗非鱼，每一群体内个体间均存在ｍｔＤＮＡ酶切片段长度多态现象，而且群体间在ｍｔＤＮＡ水平上具有一定差异，依此构建了这两种不同地方种群尼罗罗非鱼ｍｔＤＮＡ限制性内切酶酶切图谱，建立了识别它们的分子遗传标记。 相似文献
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中国对虾皮下及造血组织坏死杆状病毒（HHNBV）DNA探针的制备及应用 总被引：5，自引：2，他引：5
ＨＨＮＢＶ是１９９３年中国对虾暴发性流行病的主要病原之一。从纯化的病毒中提取ＤＮＡ，用ＥｃｏＲｌ限制性内切酶酶切成ＤＮＡ片段，与ＰＵＣ１８体外重组，建立ＨＨＮＢＶＤＮＡ文库。从中筛选出三个重组质粒（５＃、８＃、９＃），它们所插入的片段大小分别为：２．８Ｋｂ、０．８Ｋｂ、１．６Ｋｂ，它们分别用光敏生物素标记成探针，与感染ＨＨＮＢＶ的病虾ＤＮＡ呈阳性反应，以此利用制备的ＨＨＮＢＶＤＮＡ探针的高敏感性和特异性来检测虾病。 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
12.
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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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
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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
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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