不同地理群体日本囊对虾遗传多样性微卫星DNA分析

应用微卫星DNA(SSR)标记技术对福建(FJ)、湛江(ZJ)、北海(BH)地理群体的150尾日本囊对虾个体进行遗传多样性分析.10对微卫星引物在3个群体中共检测到27个等位基因,每个位点的等位基因数为2～4个.平均多态信息含量为0.3074～0.3645,平均观测杂合度为0.2960～0.4212,平均期望杂合度为0...     

从中国对虾ESTs中筛选微卫星标记的研究

利用生物信息学方法在含有10446个中国对虾ESTs的数据库中进行微卫星序列的筛选，共发现微卫星序列229个，占整个ESTs数据库的2．19％，其中含双碱基重复序列146个和3碱基重复序列58个，分别占在ESTs数据库中发现微卫星序列总数的63．76％，和25．33％，大部分发现的微卫星序列均为Perfect形式的重复序列。根据筛选得到的微卫星序列设计并合成引物19对进行多态性检测，在有扩增产物的16对引物中，首次筛选得到8个中国对虾微卫星标记，并对这些微卫星标记进行了等位基因频率、观测杂合度、期望杂合度、PIC值等统计学指标的评价。结果表明，在8个微卫星位点上，等位基因的数目从5到15不等，等位基因长度从：165～305bp，期望杂合度和多态性信息含量分别为0．59到0．89和0．56到0．88，表明这8个中国对虾微卫星标记完全适合于遗传分析。     

河鲈微卫星引物筛选

利用GenBank中获得的近缘物种微卫星序列设计的20对引物, 对河鲈养殖群体20个个体基因组DNA 进行扩增, 发现8对引物能扩增出特异性谱带, 获得32个等位基因, 5个位点的等位基因数大于或等于4个, 多态信息含量0. 3680~ 0. 7395, 5个位点多态信息含量为高度多态( PIC>0.5)。期望杂合度范围在0. 4154~ 0. 7936,观测杂合度普遍高于期望杂合度。个体识别率范围0.180~ 0. 620, 累积个体识别率为0. 9899, 单一微卫星位点的个体识别率普遍低于0. 8, 未出现高度多态性遗传标记。非父排除率范围在0. 2537~ 0. 6185, 累积非父排除率0. 9937, 4个位点的非父排除率高于0. 5, 属于高度多态性遗传标记。     

用微卫星标记分析湘华鲮野生群体遗传多样性

为了解湘华鲮天然种质资源现状，用38对鲤科鱼类微卫星引物对其群体进行全基因组扫描，结果筛选出15对能够获得稳定扩增条带的引物。以鲮鱼养殖群体为对照群体。在15个微卫星位点中，湘华鲮多态性位点9个，对照群体6个。通过分析湘华鲮微卫星位点PCR产物的电泳图谱，共检测到40个等位基因，每个位点的等位基因数目介于1～5之间，其平均等位基因数为2．67个，平均有效等位基因数为1．47个，平均观察杂合度为0．2289，平均期望杂合度为0．2730，平均多态信息含量（PIC）为0．2440，多数Hardy-Weinberg平衡偏离指数值偏离平衡值，与对照组无显著性差异。本研究表明湘华鲮野生种群结构不合理，遗传多样性低，种质资源处于危险状态。     

仿刺参的微卫星标记

为了评价种质资源及基础生物学研究的需要，本文开发了仿刺参的微卫星标记。NCBI数据库中共有20个含有仿刺参微卫星的序列，从中选取8个设计引物，发现6个微卫星位点有多态性。不同的引物获得的等位基因数为3～9个不等，6个位点共获得了31个等位基因，每个位点平均获得5．2个等位基因。6个位点的平均观测杂合度（Ho）为0．3611，平均期望杂合度（He）为0．6402。位点AJMS004提供的多态性信息含量值较低，为0．4862；其他5个位点均在0．5以上。另外，还尝试了红海参（Parastichopus californicus）微卫星标记在仿刺参的通用性。实验结果表明，在较高的退火温度下，5对引物均能扩增仿刺参的基因组DNA并具多态性。5个位点共获得了22个等位基因，每个位点平均获得4．4个等位基因。5个位点的平均观测杂合度（Ho）为0．1733，平均期望杂合度（He）为0．4201。其中位点Psc2的多态性信息含量值最高，为0．8500。     

凉山半细毛羊徽卫星标记多态性的研究

利用微卫星技术对凉山半细毛羊核心育种群206个个体第1、2、3、9号染色体上的18个微卫星位点进行研究，检测微卫星在凉山半细毛羊中的等位基因数，计算等位基因频率（Pi）、遗传杂合度（H）和多态信息含量（PIC），分析了凉山半细毛羊微卫星DNA的多态性。实验采用的18个微卫星标记位点在凉山半细毛羊群体中，平均等位基因数为8.2778个（3～19个），平均多态信息含量为0．591（O.253～0.833），遗传杂合度均值为0．500（0.026～0.888）。表明该品种绵羊遗传多态性丰富，群体遗传变异较大，并且所选18个微卫星基因座适用于绵羊的遗传连锁分析研究。     

湘西盲高原鳅遗传多样性的微卫星分析

利用筛选的16对微卫星标记对来自于湖南湘西龙山县乌龙山3个不同洞穴的盲高原鳅群体进行遗传多样性及遗传分化分析.通过计算多态信息含量、平均杂合度、等位基因数、遗传距离、基因流、F-统计量等参数,评估各盲高原鳅群体遗传多样性和各群体间遗传分化.16个微卫星标记在3个群体中共检测出83个等位基因.每个座位检测到3～8个等位基因不等.3个群体各个多态位点的平均观测杂合度分别为0.362 5～0.946 5,平均期望杂合度为0.538 6～0.906 5.3个群体多态微卫星位点的PIC分别为0.263 2、0.231 3、0.303 5,选取的16个微卫星位点中2个为高度多态,2个为低度多态,其余为中度多态.分子变异方差分析(AMOVA)结果表明,遗传变异大部分(92.84％)来自群体内,仅有7.16％的变异来自于群体间,数据表明3个群体处于未分化状态,遗传一致性较大.     

五个斑点叉尾群体的微卫星遗传多样性分析

用8对微卫星引物对1997-2004年引进的5个斑点叉尾群体进行遗传多样性分析,计算并统计等位基因数、多态信息含量（PIC）、杂合度、遗传相似性系数、遗传距离等参数。实验显示8个微卫星位点在5个斑点叉尾群体中共检测到42个等位基因,平均期望杂合度为0.6338～0.7320,表明其遗传多样性程度处于中等偏上水平。平均多态信息含量为0.5756～0.6869,说明基因座为高度多态基因座（PIC〉0.5）。群体间遗传相似系数为0.7504～0.9203。聚类分析显示,04群体与其他4个群体的亲缘关系较远。结果表明：引进的5个斑点叉尾群体均具有较高的遗传多样性,遗传信息丰富,遗传变异大,为良好的育种材料。     

牙鲆4个选择性繁育后代群体遗传结构的微卫星分析

为了检测由3个牙鲆基础群体组合交配建立的4个选择性繁育后代群体的遗传多样性水平，本研究利用10对微卫星引物分析了其遗传结构信息。计算结果表明，等位基因数3～10个，平均等位基因数5．7个，有效等位基因数1．3571～4．5979，平均有效等位基因数2．9832，各个位点的平均观测杂合度0．2083～1．0000，平均期望杂合度0．2081～0．7956，多态信息含量0．1671～0．7501，其中中度多态位点两个，高度多态位点7个，各群体的多态信息含量从大到小依次为日本亲鱼与抗病亲鱼杂交后代群体、抗病亲鱼自繁后代群体、抗病亲鱼与黄海野生亲鱼繁殖后代群体、日本亲鱼自繁后代群体。分析结果表明，4个群体的遗传多态水平均较高，对4个选择性繁育后代群体的平均观测杂合度值进行X。检验表明，群体间遗传多样性差异不显著。日本亲鱼自繁后代群体与中国海域亲鱼繁育后代群体（抗病亲鱼自繁后代群体和抗病亲鱼与黄海野生亲鱼繁殖后代群体）的遗传距离较远，分别为0．2724和0．3310。根据群体间的遗传距离利用NJ法构建系统树，抗病亲鱼自繁后代群体和抗病亲鱼与黄海野生亲鱼繁殖后代群体聚为一支，日本亲鱼自繁后代群体和日本亲鱼与抗病亲鱼杂交后代群体聚为一支，对遗传偏离指数的分析表明群体间的遗传平衡差异很大。     

河鲈微卫星引物筛选

利用GenBank中获得的近缘物种微卫星序列设计的20对引物,对河鲈养殖群体20个个体基因组DNA进行扩增,发现8对引物能扩增出特异性谱带,获得32个等位基因,5个位点的等位基因数大于或等于4个,多态信息含量0.3680～0.7395,5个位点多态信息含量为高度多态(PIC＞0.5).期望杂合度范围在0.4154～0.7936,观测杂合度普遍高于期望杂合度.个体识别率范围在0.180～0.620,累积个体识别率为0.9899,单一微卫星位点的个体识别率普遍低于0.8,未出现高度多态性遗传标记.非父排除率范围在0.2537～0.6185,累积非父排除率0.9937,4个位点的非父排除率高于0.5,属于高度多态性遗传标记.     

Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean

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.     

Growth performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

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.     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.     

Cardiac,ventilatory and metabolic responses of two ecologically dissimilar species of fish to waterborne cyanide

Changes in heart rate, ventilatory activity and oxygen consumption were determined in trout (Salmo gairdneri) and brown bullhead catfish (Ictalurus nebulosus) during exposure to a steadily increasing concentration of waterborne cyanide selected to produce death in 8–9 hours for each species. The lethal cyanide concentration for the bullheads was an order of magnitude higher than for trout. Trout developed an immediate and gradually increasing bradycardia throughout the exposure period. Cyanide produced tachycardia in the bullhead followed by a gradual onset of bradycardia as the concentration of cyanide was raised. Pericardial injection of atropine (a muscarinic cholinergic antagonist) indicated that bradycardia in the trout was due initially to increased vagal tone but later due to the direct effect of cyanide on the heart. Hyperventilation in the trout persisted throughout the exposure period, although the rate and amplitude fluctuated and was variable between individual fish. During the last hour of exposure (highest cyanide concentration), ventilation was characterized by rapid, shallow breaths followed by a sudden respiratory arrest. The bullheads exhibited hyperventilation during the first 3 hours of exposure followed by a gradual, linear drop in ventilation rate and amplitude until death occurred. Cardiac and ventilatory responses in both species were attributed to stimulation of central and peripheral chemoreceptors by cyanide. Evidence is presented which suggests the initial response in the bullheads was due, at least in part, to gustatory stimulation by the cyanide. Oxygen consumption of the trout remained above pre-exposure levels for the majority of the test period. Oxygen consumption in the bullhead paralleled the changes in heart and ventilatory rates. Whole-body lactate levels of fingerlings of both species during cyanide exposure were measured to estimate the extent of anaerobiosis. Whole-body lactate levels were much greater in the bullheads than the trout, indicating a higher capacity for anaerobiosis, possibly due to a greater fuel supply. Overall, the trout responded to cyanide in a manner similar to that produced by environmental hypoxia whereas the bullheads experienced a gustatory stimulus which masked the hypoxia-like response.     

An overview of stress physiology of fish transport: changes in water quality as a function of transport duration

This study brings an integrated analysis about the relationship between water deterioration and its physiological consequences in live fish transport. The analysis was focused on the transport water and its deterioration, and physiological challenges imposed on the fish. Usual commercial handling procedures employed to mitigate fish stress during transport were discussed. Future topics of research for the establishment of safer fish transport protocols were proposed. Transport was classified into short (≤8 h) or long transport (>8 h). The main issue in short transports should be the prevention of water pH reduction, while in long transports it is the increase in ammonia. Plasma cortisol is the most employed marker for stress and is acutely elevated upon short episodes of transport, but remains elevated even in long‐transport events. Plasma glucose is perhaps a better marker for handling stress. Plasma lactate, pH, osmolality CO₂ and ions should be more often evaluated. Plasma Na⁺ and Cl^‐ are very useful markers of acidosis, due to their respective exchange for H⁺ and , for acid–base regulation. The establishment of species‐specific transport protocols should be preceded by such combined analyses of water and physiological parameters.     

Are hatchery‐reared abalone naïve of predators? Comparing the behaviours of wild and hatchery‐reared northern abalone,Haliotis kamtschatkana (Jonas, 1845)

Abalone populations have declined worldwide, generating interest in enhancement using hatchery‐reared individuals. In many cases, such restoration efforts have met with limited success due to high predator‐induced mortality rates. Furthermore, the mortality rates of outplanted hatchery abalone are often considerably higher than for wild individuals. This study uses northern abalone (Haliotis kamtschatkana) as a case study to determine whether hatchery‐reared abalone behave differently than their wild counterparts. In the field, outplanted hatchery‐reared abalone were significantly less responsive than wild abalone, in terms of number of abalone responding and intensity of response, to nearby movement and to physical contact with an inert probe. Also, when encountering a cue to which all abalone responded (a seastar predator), hatchery‐reared individuals remained subdued. Anti‐predator behavioural deficits in hatchery‐reared abalone were more pronounced in 4‐year‐old individuals than in 1‐year‐old individuals, suggesting an influence of either age or amount of time spent in the hatchery environment. These behavioural differences are expected to increase the vulnerability of hatchery‐reared abalone to predators, and are likely a major cause of their elevated predator‐induced mortality when outplanted.     

Effects of cadmium on the kinetics of calcium uptake in developing tilapia larvae, Oreochromis mossambicus

The toxic effects of Cd²⁺ on Ca²⁺ influx kinetics in developing tilapia (Oreochromis mossambicus) larvae were evaluated. Addition of 20 µg l^-1 of Cd²⁺ to the environment of 0 and 3 day-old larvae competitively inhibited the Ca²⁺ uptake within 4h resulting in a great increase in K_m values for Ca²⁺ influx (19.3 and 17.4 fold, respectively) as compared with their respective controls. Consequently, the actual Ca²⁺ influx of larvae in solutions of 0.2 mM Ca²⁺ are suppressed by 32–45%. Also, 3 day-old larvae were more sensitive to internally accumulated Cd²⁺ than 0 day-old larvae. Although the Ca²⁺ influx in 0 and 3 day-old larvae may be restored to the levels of their respective controls with 24h of being transferred to a 20 µg l^-1 Cd²⁺ solution, total body Ca²⁺ content was significantly reduced in 3 day-old larvae. Increased Ca²⁺ uptake efficiency ensures sufficient Ca²⁺ for normal growth. However, rapid increase in Ca²⁺ influx after hatching also leads to higher Cd²⁺ uptake. Exposure to Cd²⁺ will lead to a drop in body Ca²⁺ content resulting in retardation of larval growth. Therefore, we conclude that if Ca²⁺ uptake is interfered with at this critical stage of development, larvae will not be able to maintain normal levels of body Ca²⁺ and will show signs of Cd²⁺ poisoning.     

Migratory dynamics of stream-spawning longnose gar (Lepisosteus osseus)

Abstract– Literature evidence suggests that lake-dwelling longnose gar (Lepisosteus osseus) enter tributary streams to spawn, Until the present study, the dynamics of this breeding migration had never been investigated quantitatively. During the summers of 1991 and 1992, longnose gar were captured as they entered Weaubleau Creek, Missouri, a tributary of Harry S. Truman Reservoir. The in-stream spawning migration began in early April and ended in late May, and was positively correlated with stream flow and water level, and negatively correlated with water temperature. In-stream residence times ranged from 15 to 94 days, with males exhibiting longer residence times than females. Once in-stream, longnose gar travelled as far as 10 km upstream and occupied certain pools at greater relative frequencies. Although the reason for this preferential utilization is not completely understood, it may relate to pool depth and riffle proximity. Longnose gar disperse from the spawning stream great distances, with gar captured in Weaubleau Creek being recaptured up to 48 km away. This information should provide fisheries biologists the means to consider the reproductive ecology of this species in their conservation and management decisions.     

Nutritional regulation of hepatocyte fatty acid desaturation and polyunsaturated fatty acid composition in zebrafish (Danio rerio) and tilapia (Oreochromis niloticus)

The desaturation and elongation of [1-¹⁴C]18:3n-3 was investigated in hepatocytes of the tropical warm freshwater species, zebrafish (Danio rerio) and Nile tilapia (Oreochromis niloticus). The hepatocyte fatty acid desaturation/elongation pathway was assayed before and after the fish were fed two experimental diets, a control diet containing fish oil (FO) and a diet containing vegetable oil (VO; a blend of olive, linseed and high oleic acid sunflower oils) for 10 weeks. The VO diet was formulated to provide 1% each of 18:2n-6 and 18:3n-3, and so satisfy the possible EFA requirements of zebrafish and tilapia. At the end of the dietary trial, the lipid and fatty acid composition was determined in whole zebrafish, and liver, white muscle and brain of tilapia. Both zebrafish and tilapia expressed a hepatocyte fatty acid desaturation/elongation pattern consistent with them being freshwater and planktonivorous fish. The data also showed that hepatic fatty acid desaturation/elongation was nutritionally regulated with the activities being higher in fish fed the VO diet compared to fish fed the FO diet. In zebrafish, the main effect of the VO diet was increased fatty acid Δ6 desaturase activity resulting in the production of significantly more 18:4n-3 compared to fish fed the FO diet. In tilapia, all activities in the pathway were greater in fish fed the VO diet resulting in increased amounts of all fatty acids in the pathway, but primarily eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3). However, the fatty acid compositional data indicated that despite increased activity, desaturation of 18:3n-3 was insufficient to maintain tissue proportions of EPA and DHA in fish fed the VO diet at the same level as in fish fed the FO diet. Practically, these results indicate that manipulation of tilapia diets in commercial culture in response to the declining global fish oil market would have important consequences for fish fatty acid composition and the health of consumers. Scientifically, zebrafish and tilapia, both the subject of active genome mapping projects, could be useful models for studies of lipid and fatty acid metabolism at a molecular biological and genetic level. This revised version was published online in August 2006 with corrections to the Cover Date.     

Protein and amino acid composition from the mantle of juvenile O ctopus vulgaris exposed to prolonged starvation

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.