太湖似刺鳊(鱼句)染色体组型分析及细胞核DNA含量

采用PHA、秋水仙素碱腹腔或背部肌肉注射,活体培养法,用空气干燥法制片,运用Micromeasure version 3.3染色体分析软件和Photoshop 7.0对似刺鳊(鱼句)染色体数目和核型进行了分析;同时,取似刺鳊鱼句血细胞、尾鳍、鳃、肌肉、性腺和肝脏为材料,以鸡血细胞为DNA标准(2.50 pg/2c),使用EPICS-XL型流式细胞仪测定了似刺鳊(鱼句)细胞核DNA含量.结果显示:染色体众数2n=50,染色体大小的绝对值为0.86～2.32 μm,平均长度为1.63 μm,未观察到次缢痕及异配型染色体,亦未发现有随体,核型公式为18m+20sm+8st+4t,NF=88.6个组织细胞核DNA含量分别为:尾鳍3.779 pg/2c,鳃4.007 pg/2c,肌肉3.819 pg/2c,性腺(卵巢4.242 pg/2c、精巢1.842 pg/2c),肝脏3.905 pg/2c.尾鳍、鳃、肌肉和肝脏的细胞核DNA含量不存在显著差异(P＞0.05),但都极显著高于血细胞DNA含量(P<0.01);卵巢细胞核DNA含量约为精巢的2倍.     

白乌鳢染色体核型和DNA含量分析

通过向活体白乌鳢(Opniocepnalus argus)腹腔注射植物血凝素(PHA)和秋水仙素进行短期药物处理后,制备染色体标本并对其核型进行分析。以鸡血细胞DNA含量为标准(2.50 pg),用流式细胞仪测定了白乌鳢外周血细胞的DNA含量。结果表明:(1)白乌鳢的染色体组为2 n=48,核型公式为4 m+22 st+22 t,总臂数NF=52,无性染色体。(2)白乌鳢的DNA含量为鸡血对照的0.59倍,其DNA含量为(1.48±0.04)pg/N。白乌鳢的染色体数和DNA含量均显示其具有二倍体特征。依据实验结果推论白乌鳢为乌鳢的白化变异个体。     

银鲳亲鱼不同组织的氨基酸及其随性腺发育的变化

测定了海捕银鲳亲鱼肌肉,肝脏和性腺组织的氨基酸,并分析了不同性腺发育阶段的氨基酸变化。结果表明,雄性V期亲鱼精巢中的牛磺酸含量最高（27.8mg/g）,显著高于肌肉和肝脏组织,而雄性V期亲鱼氨基酸总量在肌肉中最高（631.4 mg/g）,其次是精巢（544.8 mg/g）和肝脏（468.0 mg/g）组织。雌性亲鱼肝脏中的牛磺酸含量（13.7 mg/g）显著高于卵巢和肌肉组织。在卵巢从III期发育到V期过程中,雌性亲鱼肌肉和肝脏中的氨基酸总量在V期下降显著,而卵巢氨基酸总量维持稳定。在总氨基酸的组成上,肌肉中含量高的氨基酸为Glu,Lys, Asp,Leu和Arg;肝脏中含量高的为Glu,Lys, Asp,Val和Leu;精巢中含量高为Glu,Asp, Lys, Arg和Val;卵巢含量高中为Glu,Lys,Leu,Val和Asp。性别和性腺发育阶段对亲鱼肌肉总氨基酸组成无影响。游离氨基酸含量在III 、IV和 V期卵巢中的含量分别为17.7 mg/g ,41.5 mg/g和29.6 mg/g,且其组成随性腺发育阶段的不同而有显著差异。     

杂交鲌“先锋1号”染色体组型分析和DNA含量测定

给体质量22～88 g的杂交鲌"先锋1号"[翘嘴红鲌(♀)×黑尾近红鲌(♂)杂交子一代(F_1)]幼鱼胸鳍基部注射6μg/g的植物血球凝集素,12～14 h后再注射8μg/g秋水仙素,2 h后尾静脉采血,经常规空气干燥、Giemsa染色、显微拍照及Photoshop CS 6软件分析等步骤研究"先锋1号"F_1的染色体数目、核型等;用公鸡血细胞DNA绝对含量(2.50 pg/2C)作对照,用流式细胞仪测定"先锋1号"F_1血细胞DNA含量,以丰富其细胞遗传学特征。试验结果显示,杂交鲌"先锋1号"的染色体数目为2n=48,核型公式为28m+18sm+2st,染色体臂数NF=94,未发现次缢痕和随体,也未发现与性别有关的异型性染色体;杂交鲌"先锋1号"的DNA含量为(3.16±0.04) pg/2C。杂交鲌"先锋1号"的染色体数目与父母本相同,核型与父本黑尾近红鲌更接近,DNA含量与父母本无显著性差异。     

中国胭脂鱼的细胞遗传学分析

采用植物血凝素、秋水仙素活体注射和肾细胞直接制片法分析了中国胭脂鱼(Myxiocyprinus asiaticus)的染色体核型和Ag-NORs显带;并以中国胭脂鱼血细胞为样本,鸡血细胞DNA含量为标准(2.50 pg),用流式细胞仪测定了中国胭脂鱼的 DNA 含量.结果表明:(1)中国胭脂鱼染色体数目2n=100,核型公式为10 m+4 sm+12 st+74 t, NF=114,未观察到次缢痕及随体,亦未发现有性染色体.(2)中国胭脂鱼染色体具有1对Ag-NORs,位于第1对染色体短臂上,间期核中Ag-NORs的数目为1~2个,未见Ag-NORs的联合现象.(3)PI和DAPI作为荧光染料,得出中国胭脂鱼DNA含量分别为5.50±0.24 pg和6.16±0.03 pg.(4)中国胭脂鱼为四倍体,在特定的分类阶元中属于较原始的鱼类.     

闽东海域银鲳亲鱼性腺发育后期脂类及脂肪酸蓄积特点

脂肪和脂肪酸是海水鱼类早期生长发育的重要结构物质和能量来源。亲鱼的脂肪和脂肪酸储备影响其繁殖性能和早期仔鱼的发育。为了获知银鲳亲鱼性腺发育后期脂类及脂肪酸蓄积特点,本研究采用氯仿甲醇法及气相色谱法定量检测了繁殖季节闽东海域野生银鲳亲鱼不同组织的总脂肪及脂肪酸含量。结果表明:亲鱼卵巢、精巢、肝脏和肌肉的总脂肪含量差异显著。卵巢、精巢、肝脏和肌肉的总脂含量分别为:35.76%,15.11%,22.07%和22.14%(占组织干重)。极性脂肪占总脂肪的比例在精巢中最高,其次为肝脏和卵巢,在肌肉中最低。性腺从Ⅳ期发育到Ⅴ期,雌鱼卵巢总脂肪和中性脂肪含量显著增加,雄鱼肌肉极性脂肪含量显著降低。中性脂肪中卵巢的20∶5n-3(EPA,2.25~3.87 mg/g)、22∶6n-3(DHA,6.71~13.03mg/g)和高不饱和脂肪酸(HUFAs,17.20~29.64 mg/g)含量最高,极性脂肪中精巢的EPA(0.38~0.27 mg/g)和DHA(3.12~3.59 mg/g)含量最高。性腺中n-3/n-6比值显著高于肝脏和肌肉中。随着卵巢发育,DHA等必需脂肪酸在雌鱼不同组织及同一组织不同脂肪类别之间存在转移现象。研究表明,银鲳亲鱼各组织的总脂肪含量、总脂肪组成及脂肪酸绝对含量(mg/g干物质)具有组织特异性,随着性腺发育,必需脂肪酸总体上表现为由肌肉和肝脏向性腺中转移,且性腺中脂肪酸的变化主要发生在中性脂肪中。     

小体鲟染色体核型及倍性的细胞遗传学研究

采用体内注射PHA和秋水仙素,肾细胞短期培养,常规空气干燥法制备小体鲟(Acipenser dabryanus Dumerii)的染色体,对肾细胞染色体数目统计分析表明,小体鲟染色体组是由116±4条染色体组成,核型公式为40m+34sm+26st+16t±mc,NF 190±;采用德国Partec pasⅢ型流式细胞分析仪,以鸡红细胞为标准DNA(含量为2.3pg/N),测定小体鲟体细胞DNA含量为6.06 pg/N,与染色体分析结果比较倍性一致.综合以上两项结果可以得出,小体鲟为四倍体鲟鱼.     

中华绒螯蟹不同组织中基因组DNA含量比较

用改进后的柱式回收法提取了中华绒螯蟹不同组织中的基因组DNA,并通过紫外吸收光谱法初步分析了附肢肌肉组织、鳃、性腺组织和肝脏组织中基因组DNA的含量,初步得出同质量不同组织中基因组DNA含量大小关系趋向于:鳃>肝脏>性腺>附肢肌肉.     

青蛤染色体制备及核型分析

以青蛤成体鳃组织和性成熟的雌、雄青蛤性腺为材料,采用常规制片方法,分别制备青蛤二倍体染色体和单倍体染色体,统计染色体数目。结果显示,青蛤二倍体与单倍体染色体数目分别为2n=38和n=19;核型分析发现,青蛤核型组成为11m+6sm+2st,NF=76,未发现性染色体和随体染色体。本研究以青蛤成熟性腺为制备材料,成功制备单倍体染色体,与青蛤二倍体染色体互相印证,弥补了以往研究的不足,进一步丰富了对青蛤染色体数目及核型组成的认知。同时,也为青蛤染色体水平全基因组测序组装提供更加科学的基础资料。     

团头鲂核型与DNA含量分析研究

采用PHA体内培养法，取鱼体肾细胞用空气干燥法制备团头鲂染色体标本。经核型分析，团头鲂的核型为2n=48,26m 18sm 4st，臂数（NF）为92。用流式细胞仪测量外周血红血球细胞核的DNA含量为2．66pg/N。实验结果分析表明团头舫染色体数及DNA含量是稳定的，其核型公式与有关文献的报道略有差异。     

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.     

Metabolic responses of matrinxã, Brycon amazonicus (Spix & Agassiz, 1829), exposed to environmental nitrite

Nitrite is usually found in aquatic environments where nitrification process occurs. This ion can cause several injuries to aquatic species, particularly fish. Nitrite reacts with haemoglobin yielding the non‐functional methaemoglobin, which leads to many physiological consequences such as functional anaemia and supposed hypoxia. Metabolism of the freshwater teleost matrinxã, Brycon amazonicus, exposed to environmental 0.6 mg L^?1 of nitrite N‐NO₂^? for 96 h was studied, and the fermentative/oxidative preference was gauged. Concentrations of glycogen, glucose, lactate, pyruvate and ammonia, plus the activities of lactate, glutamate and malate dehydrogenase were assayed. The exposure resulted in a metabolic profile that allowed inferring the continuity of oxidative metabolism. Catabolism of amino acids prevailed or was apparently exacerbated by inferred branchial injury and consequent impairment of nitrogen excretion. Moreover, the studied enzymes glutamate dehydrogenase, lactate dehydrogenase and malate dehydrogenase from brain and heart were little affected by nitrite. The expected fermentative metabolism due to the high methaemoglobin formation was not observed.