Effects of dietary levels of protein on growth,feed utilization and physiological parameters for juvenile Dabry's sturgeon,Acipenser dabryanus

Five semi‐purified diets were formulated to contain graded levels of dietary protein (340, 390, 440, 490 and 540 g/kg diet), and each diet was assigned to triplicate groups of eight juvenile Dabry's sturgeon for 50 days. Results showed that specific growth rate increased linearly with increasing dietary protein levels from 340 to 440 g/kg diet, then levelled off at higher protein levels. Higher dietary protein (440 and 540 g/kg) significantly increased the feed intake and feed efficiency ratio of fish compared to lower protein diets (340 to 490 g/kg diet) (p < .05). Fish fed a 440 g protein kg^?1 diet had higher protein efficiency ratio and serum lysozyme activity than other treatments. Serum ammonia content and activities of liver aminotransferase were positively correlated with dietary protein levels. No significant difference (p > .05) among groups was observed in glycogen content. As dietary protein level increased, protein and ash content of fish dorsal muscle were greatly enhanced, whereas lipid content was significantly reduced (p < .05). Based on broken‐line regression analysis of SGR against dietary protein levels, the optimum dietary protein level for maximal growth of juvenile Dabry's sturgeon is 446.68 g/kg diet.     

Genetic diversity of Nile tilapia populations revealed by randomly amplified polymorphic DNA (RAPD)

Genetic diversity of the Nile tilapia Oreochromis niloticus collected from the river Nile (Cairo, Assuit and Qena) and two Delta lakes (Burullus and Manzalla) in Egypt was examined by the analysis of randomly amplified polymorphic DNA (RAPD). Of 25 primers examined,21 primers produced 230 RAPD bands. The percentage of polymorphic bands in Manzalla (29.4%) and Burullus (24%) populations was low compared with Assuit (30.54%), Cairo (33.5%) and Qena (44.84%) populations. The highest percentage of polymorphic bands was observed in the Qena population,suggesting a greater potential for use in breeding programs. The molecular phylogenetic tree constructed by unweighted pair‐group method of analysis shows Manzalla and Burullus populations strongly linked and separate from the Assuit and Cairo populations, with Qena population as outgroup. The data serve as a baseline analysis of the current genetic diversity found among O. niloticus populations in Egypt.     

Microsatellite analysis of genetic diversity and genetic structure of the Chinese freshwater mussel Solenaia carinata (Bivalvia: Unionidae)

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3个仿刺参地理种群遗传变异的微卫星DNA分析

运用微卫星DNA技术对仿刺参烟台群体、威海群体、大连群体的3个野生群各20个个体进行了遗传分析。对9个基因位点进行了扩增,共获得了59个等位基因。评估了9对微卫星引物的多态信息含量(PIC),范围在0.5129~0.8794之间。结果表明:3个野生群体的平均杂合度观测值分别为0.6416、0.6595、0.5824,平均杂合度期望值分别为0.7641、0.7161、0.7364;在Hardy-Weinberg平衡条件下,进行了P检验,发现3个群体均有位点发生了显著偏离;对3个群体进行了配对Fst值计算,发现3个群体之间遗传分化较弱。从变异贡献率来看,95.68%的变异来自个体之间,4.32%的变异来自群体之间。经过聚类分析,发现威海群体和大连群体之间亲缘关系较近,烟台群体与前两群体亲缘关系较远。     

Inheritance of microsatellite loci in polyploid Siberian sturgeon (Acipenser baeri Brandt) based on uniparental haploids

Haploid gynogenesis was induced in Siberian sturgeon (Acipenser baeri Brandt) using UV‐irradiated bester (hybrid of Huso huso L. and Acipenser ruthenus L.) sperm. The inactivated heterologous sperm was used successfully to produce gynogenetic haploids of Siberian sturgeon. All haploid progeny were screened using microsatellite DNA analysis and uniparental transmission in gynogenetic haploids was confirmed. The objective of this study was to obtain haploids of Siberian sturgeon and describe the inheritance of microsatellite loci in haploid embryos of this species. This initial work, based on inheritance in haploid Siberian sturgeon, suggests that three studied microsatellite loci segregate disomically, three tetrasomically and one octasomically.     

Evaluation of lethal and non-lethal sampling methods for the detection of white sturgeon iridovirus infection in white sturgeon, Acipenser transmontanus (Richardson)

Pectoral fin tissue of white sturgeon was investigated as a potential non-lethal sample source for the detection of white sturgeon iridovirus (WSIV) infection. Histopathology and polymerase chain reaction (PCR) results using fin tissue were compared with the standard lethal histopathology sampling method that utilizes head tissue. Tissues for each of the three sampling methods were collected weekly for 8 weeks from individual sturgeon undergoing an experimental cohabitation challenge with fish infected with the Abernathy isolate of WSIV. Non-lethal fin histopathological evaluation did not reveal infection during the first 3 weeks of sampling, while non-lethal PCR and the lethal method were variable. However, all three sampling methods were equally capable of identifying infection from 4 to 8 weeks post-exposure. Of the survivors tested, all were negative by PCR and the lethal method, and only one fish was identified as being positive by non-lethal fin histopathology. In another experiment, all three sampling methods were applied to asymptomatic WSIV carriers in a case study conducted at the Kootenai Tribal Sturgeon Conservation Hatchery. Results showed that both lethal and non-lethal fin histopathology were equally effective in detecting infection, but PCR was unable to identify this strain of WSIV. Depending on the virus isolate, these results suggest that non-lethal sampling of fin tissue (histopathology or PCR) is comparable with the lethal sampling method at identifying WSIV infection once infection is established, and under certain circumstances may provide an alternative to lethal sampling.     

运动功能相关的中华鲟和西伯利亚鲟幼鱼侧视形态比较研究

通过选取11个侧视形态特征度量进行主成分分析(principal component analysis,PCA),同时设置26个标记点进行相对扭曲分析(relative warp analysis,RWA),比较了中华鲟(Acipenser sinensis)和西伯利亚鲟(A.baerii)幼鱼在侧视形态上的差异以及相对扭曲分析和传统多元分析在鲟鱼形态研究中的优劣。结合前期对它们的有氧游泳能力比较结果(西伯利亚鲟比中华鲟强25%),分析了鲟鱼形态的水动力功能。主成分分析比较表明,中华鲟头长、头高、背鳍前基点之后的躯干高度、尾柄长、背鳍前缘长度均显著大于西伯利亚鲟(P<0.05),而尾鳍上叶显著短于西伯利亚鲟(P<0.05)。相对扭曲分析计算样本的几何信息并可视化统计结果,表明中华鲟的吻厚、吻长、头高、头长、躯干后半段高、背鳍前缘长显著大于西伯利亚鲟,而尾柄长和尾鳍上下叶长显著小于西伯利亚鲟(P<0.001)。上述形态特征直接影响着两种鲟鱼的游泳能力。     

长江上游长鳍吻鮈群体线粒体控制区遗传多样性分析

分析长鳍吻鮈群体遗传结构和遗传多样性,为研究过度捕捞、水电开发等人类活动对长鳍吻鮈的长期生态学效应及其物种适应机制提供基础数据,同时也为该物种保护策略的制定和调整提供科学数据支持。2014和2015年,于长江上游干流江津、宜宾、皎平渡和格里坪采集长鳍吻鮈,利用线粒体控制区序列的特异性引物,通过PCR扩增以及序列测定,获得125份长鳍吻鮈线粒体控制区序列。结果表明:125尾长鳍吻鮈样品共检测到多态位点25个,单倍型30种,平均单倍型多样性指数(Hd)和核苷酸多样性指数(Pi)分别为0.785和0.00140;中性检验(Tajima's D为-1.97524,P0.05;Fu's Fs为-31.374,P=0.000)和序列错配分布结果表明,长江上游长鳍吻鮈历史上经历过"遗传瓶颈"和种群扩张;分子方差分析(AMOVA)结果表明,群体间的遗传变异绝大部分来源于群体内部(99.07%),群体间无显著遗传分化(Fst=0.00933,P0.05);单倍型网络、Kinura 2-Parameter遗传距离和平均基因流均显示,4个长鳍吻鮈群体间存在广泛的基因交流,群体间无遗传分化。各群体间没有显著遗传分化,基因交流频繁,可将长江上游长鳍吻鮈作为单一的进化显著单元(ESU)进行管理。     

从连续到偶发: 中华鲟在葛洲坝下发生小规模自然繁殖

归纳了白鲟、长江鲟、中华鲟近40年自然资源量变化情况,统计了长江鲟和中华鲟人工增殖放流数据,评估了增殖放流成效。因物种特性和分布的差异,放流无效的原因迥异。过度捕捞使人工放流长江鲟在放流后6个月之内难逃被“误捕”,而中华鲟放流数量不足及长江和近海过度捕捞导致中华鲟人工增殖放流的贡献甚微,其结果是长江鲟和中华鲟增殖放流均无法达到自然繁殖群体的补充水平。本文探究了近40年来白鲟、中华鲟和长江鲟在保护、管理和决策上存在的误区和不足,提出在生态大保护的背景下,① 应编制和实施《长江鲟拯救行动计划》优先项目;② 应以《中华鲟拯救行动计划》为指引,在就地保护、迁地保护和遗传多样性保护方面提出更具有可操作性的优先计划;③ 应设立中华鲟和长江鲟物种拯救行动计划专项,对现有涉栖息地或保护区生态补偿项目进行优化整合。唯有以恢复长江鲟和中华鲟自然繁殖为核心,才有望延续和恢复其自然种群,实现人与自然的和谐共处,推动长江流域社会经济绿色可持续发展。     

Genetic diversity in the Indian population of Penaeus monodon (Fabricius, 1798) as revealed by mtDNA sequence analysis

Genetic diversity in the Indian population of the tiger shrimp Penaeus monodon was determined by using partial sequence data of the mitochondrial DNA (mtDNA) control region (D‐loop) and the 16S rRNA gene. Eight populations from different geographical locations (Mumbai, Kochi, Mangalore, Kakinada, Gopalpur, Chilika, Paradeep and Andaman) were collected and analysed. The amplified polymerase chain reaction products of size 577 bp for the control region and 472 bp for 16S rRNA were sequenced in both directions and data were analysed through clustal , arlequin , mega and phylip . A significant genetic structure was found among the Indian populations. The mtDNA control region proved to be a powerful marker in comparison with 16S rRNA for population studies of this species. The east coast population was more genetically diverse than the west coast. The Andaman population was found to be the most diverse among all the populations. The populations on the west coast were found to be genetically more structured and differentiated than the populations on the east coast. The results revealed a high level of genetic diversity and also distinct population structuring of P. monodon, suggesting great possibilities of genetic improvement for growth and other economic traits.     

Induction of gynogenesis in stellate sturgeon (Acipenser stellatus Pallas, 1771) and its verification using microsatellite markers

Normal egg fertilization was carried out in stellate sturgeon Acipenser stellatus sperm by 60‐, 90‐ and 120‐s ultraviolet irradiation, and a cold shock was carried out afterwards. The treatments included haploids, diploid gynogens, triploid and control groups. Fertilization, hatching and survival rate at the 60‐s irradiation time were significantly greater than those in the 90‐s and 120‐s irradiation and control groups. Two pairs of microsatellite markers were used to find gynogenesis in comparison with the parental and offspring genome in one locus. The results showed all maternal genomes among gynogen offspring with no paternal genome. The markers also showed the maximum gynogenesis induction in the Gy60 treatment.     

长江口中华鲟幼鱼主要饵料生物体内重金属Cu、Cd和Hg的积累与评价

为评价重金属污染对长江口中华鲟幼鱼生长的影响,在长江口水域采集了日本沼虾(Macrobraohium nipponense)、中国毛虾(Acetes chinensis)、狼牙鳗虾虎鱼(Taenioides rubicundus)、矛尾虾虎鱼(Chaeturichthys stigmatias)、窄体舌鳎(Cynoglossus gracilis)等5种中华鲟幼鱼重要饵料生物,研究了Cu、Cd、Hg3种重金属元素在这些饵料生物体内的积累特征,评价了饵料生物受重金属污染的程度。结果显示,虾类Cu的单项污染指数(Pi)均大于0.5,鱼类Cu的Pi均小于0.5;5种饵料生物Cd和Hg的Pi均大于1。表明它们均不同程度受到重金属污染,虾类的重金属污染程度比鱼类严重。与历史资料比较发现,长江口鱼虾的重金属污染存在加重趋势,重金属污染已对洄游入海的中华鲟幼鱼构成威胁。     

Effect of chloride on nitrite‐induced methaemoglobinemia in Atlantic sturgeon,Acipenser oxyrinchus oxyrinchus (Mitchill)

We evaluated the effects of chloride concentration on the clinical pathology in juvenile Atlantic sturgeon, Acipenser oxyrinchus oxyrinchus (Mitchill), following semi‐static exposures to 1 mg L^?1 nitrite for 96 h. In spring water naturally low in chloride (5 mg L^?1), plasma nitrite concentrated to more than 40× environmental levels resulting in a severe methaemoglobinemia characterized by torpid behaviour, 30‐fold increase in methaemoglobin fraction, anaemia, leucopenia and hyperkalaemia. Loss of intracellular water and potassium to extracellular space may have resulted in hyperkalaemia and haemodilution. Fish survived nitrite exposure, but 60% of torpid fish died following capture and tissue sampling. Fish acclimated to 10‐fold higher chloride content (55 mg L^?1) did not concentrate nitrite in the plasma above environmental levels or develop methaemoglobinemia, but did exhibit similar haematology and plasma chemistry changes. Plasma nitrite returned to preexposure levels by 14 days following nitrite exposures, but severity of clinical pathology changes persisted or increased, suggesting that Atlantic sturgeon have reduced capacity to recover from methaemoglobinemia. Fish that survive methaemoglobinemia may be susceptible to mortality from the cumulative effects of intoxication, handling and other stresses for two or more weeks following nitrite remediation. Chloride buffering in aquaculture systems reduces the toxic effects of nitrite accumulation.     

DNA fingerprinting: application to conservation of the CITES-listed dragon fish, Scleropages formosus (Osteoglossidae)

Since 1975,CITES has listed the dragon fish, Scleropages formosus, as anendangered species. In 1995, a captive-bred population was set upby a commercial fish farm with assistance from the PrimaryProduction Department in Singapore. Other farms in Indonesia andMalaysia followed suit. These populations have contributed to animmediate conservation of the species. Due to very high demandfor this ornamental fish, these venues may be its last sanctuary.DNA fingerprints of the dragon fish were obtained by different methods from the green, red and gold varieties grown in a Singapore fish farm to determine which method was most suitable in providing information on genetic variability. Because a DNA fingerprint is a pattern made up of DNA fragments that are resolved by electrophoresis, each individual has its own unique fingerprint due to a genetic make-up different from another individual. Thus, genetic variability was best studied by developing DNA fingerprints.Firstly, restriction fragment length polymorphisms (RFLPs) were obtained. DNA fragments formed by cleavage with nine restriction endonucleases used singly were hybridized individually to four non-radioactively labelled probes to give RFLPs. The RFLPs for each variety were similar and genomic DNA from each variety had many binding sites to the probes. This made differentiating RFLPs specific to individual varieties difficult. Secondly, random amplified polymorphic DNA (RAPD) fingerprints were developed. DNA fragments that were resolved on a denaturing polyacrylamide gel were hybridized to seven arbitrary primers used singly. RAPD fingerprints for each variety were different for each primer tested. The similarity index indicated low genetic variability between varieties. Lastly, DNA was screened for microsatellite loci which refer to short tandem repeats of two or three bases. The occurrence of other microsatellite loci, their chromosome location and frequency is being investigated while primers have been designed to detect more loci by the polymerase chain reaction. As this method provides undisputed and reproducible evidence of relatedness and stock identification, and can be applied for long-term management of domesticated populations through pedigree construction and evaluation of heterozygosity, it is the preferred choice to determine genetic variability     

Impact of starvation on digestive enzymes activities and plasma metabolites in Siberian sturgeon (Acipenser baerii,Brandt, 1869)

To increase the current knowledge about the relationship between nutritional status and the digestive capacity of Siberian sturgeon (Acipenser baerii), we addressed the effect of starvation‐refeeding and macronutrient composition on growth parameters and key digestive enzyme activities in A. baerii. Acipenser baerii juveniles were fed four different diets for 3 weeks, then starved for 2 weeks and allowed to refed for 5 weeks with the same diets. Another group of fish were fed 10 weeks with the corresponding diets. Among 10‐week fed fish, high‐protein diets promoted higher body weight values, while the lowest specific growth rate was observed in fish fed a low‐protein, medium‐carbohydrate, high‐lipid diet (p < .05). At the end of the experiment, in fish refed for 5 weeks following a feeding‐starvation cycle and in 10‐week fed animals, the higher levels of blood glucose, triglycerides and cholesterol were found in fish fed low‐protein diets (p < .05). In all treatments, 2 weeks of starvation decreased α‐amylase activity in the intestine (p < .05), while 4 days of refeeding increased lipase (p > .05) and α‐amylase activity in the intestine as well as pepsin in the stomach (p < .05). Our findings suggest that A. baerii maintains a high capacity to digest proteins and lipids after 2 weeks of starvation and that α‐amylase can be used as an indicator of the nutritional status in fish submitted to starvation‐refeeding cycles. Indeed, refeeding with high‐protein and CHO:L ratio diets after starvation could improve the growth rate of A. baerii in culture.     

Effect of visible implant elastomers on the growth,survival and tag retention in juvenile Atlantic sturgeon (Acipenser oxyrinchus) in laboratory conditions

This study identifies the possibility of using visible implant elastomers (VIE) to tag juvenile Atlantic sturgeon (Acipenser oxyrinchus). The mean body weight was 2.4 ± 0.4 g, and mean body length 5.5 ± 0.5 cm. Markers in three colours were used: blue, orange and green. Implant elastomers were injected in the rostrum and in the tail shaft. The experiment was conducted over a period of 10 weeks and included also a control group, that is, no tagging. No significant effect of tagging on the specific growth rate, feed conversion ratio and condition factor of juvenile Atlantic sturgeon was observed. The fish in all groups increased their body weight more than 10 times, reaching body weight of 25–28 g. The final survival in all of the groups was approximately 40%. The final tag retention varied depending on the implant colour. The highest retention of 100% was obtained with orange VIE and was significantly different from blue and green. Tag retention was dependent on the place of marking (P > 0.05). A correlation was observed between the applied dye and the place of marking (P = 0.0166). The best results were obtained when marking with orange VIE in the rostrum as all markers were visible after 70 days even without the use of UV light. According to our research, marking with VIE does not negatively affect growth and survival of juvenile Atlantic sturgeon and can be recommended for fish with an initial body weight of approximately 2.5 g in the case of short‐term research.     

Ascorbic acid status as affected by dietary treatment in the Siberian sturgeon (Acipenser baeri Brandt): tissue concentration,mobilisation and L-gulonolactone oxidase activity

A study was conducted to evaluate tissue storage and mobilisation of L-ascorbic acid (AA) in the Siberian sturgeon (Acipenser baeri) fed three different experimental diets. The three treatments consisted of a diet devoid of vitamin C (diet A0) and two diets supplemented with equivalent of 300 mg AA kg^–1 in the form of either silicone-coated ascorbic acid (diet SC) or of ascorbyl-2-polyphosphate (diet AP). During the first phase (4 months) of the trial, six batches of 130 Siberian sturgeon (initial body weight: 25.5±0.5 g) each were fed one of the three diets in duplicate. During the second phase (3 months), fish from groups SC and AP were switched to diet A0 and those fed diet A0 during the first phase were switched to diet SC. Irrespective of the dietary treatment, growth rates were not significantly different from each other. At the end of phase I, in all tissues studied, total ascorbic acid (TAA) concentrations were higher in Siberian sturgeon fed diet AP than in the other two groups. During phase II, tissue ascorbate depletion was also higher in the AP group than in the other two groups. Transfer of the AA-free diet fed group onto a diet supplemented with 300 mg AA kg^–1 (diet SC) led to a slight increase in the TAA concentrations in all tissues. Blood plasma tyrosine concentrations were not significantly different between the three groups. Whole-body collagen levels were affected by dietary AA levels or forms at the end of phase I; the differences were not significant at the end of phase II. Muscle collagen levels were slightly affected. L-Gulonolactone oxidase activity was found in the kidney of Siberian sturgeon, but not in the liver. The ascorbyl-2-polyphosphate appears to be either better utilised by Siberian sturgeon, like in many other teleosts, or more stable than the silicone-coated AA during food processing and storage. Presence of L-gulonolactone oxidase activity in Siberian sturgeon kidney combined with the absence of gross scorbutic signs in AA-free diet fed groups expressing very good growth rates suggested no need of dietary AA byA. baeri.     

中华鳖3个地理群体线粒体基因D-loop区遗传多样性分析

采用PCR结合DNA测序技术和SSCP技术,分析了中华鳖3个地理群体线粒体DNA(mtDNA)D-loop区部分序列的变异及遗传多样性。在25个个体中,其碱基组成为A+T的平均含量(65.4%)高于G+C(34.6%),共检测到变异位点7个(约占总位点数的5.7%),转换/颠换值为2.76。核苷酸多样性(π)为0.019 95,平均核苷酸差异数(K)为2.453。25个个体分属6个单倍型,单倍型多样度(Hd)为0.707,单倍型间的平均遗传距离(P)为0.027。6个单倍型构建的UPGMA系统树聚为3个分支。结果表明,中华鳖群体mtDNA D-loop区序列存在着较丰富的变异和遗传多样性,日本鳖的遗传多样性比黄河鳖和黄沙鳖丰富,黄沙鳖与日本鳖、黄河鳖的遗传距离较远。而且PCR-SSCP可以检测到两种类型的电泳图谱,其中Ⅱ型均为日本鳖,该技术可用于78位TT←→AA颠换变异位点的检测。     

长江宜宾江段圆口铜鱼遗传多样性的微卫星分析

选用实验室克隆的23个圆口铜鱼(Coreius guichenoti Sauvageet Dabry)微卫星标记分析了长江宜宾江段的圆口铜鱼群体遗传多样性,统计分析了有效等位基因数、观测杂合度Ho、期望杂合度He、多态信息含量(PIC)等遗传学指标。结果表明:23个位点有14个微卫星位点呈单态,9个位点出现多态,在这9个位点中共检测到48个等位基因,其平均有效等位基因数为5.3,多态信息含量在0.440～0.839之间变动,平均为0.670,除YT17和YT22位点属于中度多态外,其余7个位点均属于高度多态。平均观测杂合度为0.753,平均期望杂合度为0.728,表明该群体的遗传多样性较为丰富。     

The effect of dietary taurine on growth performance and liver histopathology in Persian sturgeon,Acipenser persicus (Borodin, 1897) fed plant‐based diet

This experiment was conducted to illustrate the effect of dietary taurine levels on Persian sturgeon, Acipenser persicus (mean weight of ~35 g) fed plant‐based diet. A basal diet containing low level of fishmeal (19%) was formulated containing 0.05% endogenous taurine (control diet). To this diet were added five levels of exogenous taurine: 0.25 (0.25T), 0.5 (0.5T), 0.8 (0.8T), 1.2 (1.2T) and 1.6% (1.6T). These diets were fed to the experimental fish for 42 days. Results showed that growth performance decreased and food conversion ratio increased as dietary taurine increased, although no significant difference was observed between the control and 0.25T treatments. Increased dietary taurine led to increased carcass moisture and decreased carcass lipid percentages. Feed intake was significantly and negatively correlated with dietary taurine. Carcass taurine content significantly increased with dietary taurine elevation up to 1.2% and then decreased. Hepatic taurine content showed a significant increase in 0.25T treatment and then decreased along with dietary taurine elevation and reached the control values at 1.6% taurine level. Dietary taurine elevation led to more hepatic damages compared to the control. In conclusion, although within the range reported for other studied fish, the tested taurine levels in this study might be supra‐optimal causing adverse effects in Persian sturgeon and further study with lower taurine levels is encouraged.