液相色谱-三重四级杆复合线性离子阱质谱法分析恩诺沙星在鲤鱼(Cyprinus carpio)中的代谢产物

采用液相色谱-三重四级杆复合线性离子阱质谱(HPLC/Q-TRAP MS),在正离子检测模式下,对鲤鱼(Cyprinus carpio)腹腔注射恩诺沙星给药后肝脏中的代谢产物进行分析。根据保留时间和各色谱峰质谱裂解规律,推测了恩诺沙星在鲤鱼肝脏中的代谢产物,同时,根据二级质谱碎片离子推测代谢途径及代谢产物结构。结果显示,恩诺沙星进入鲤鱼肝脏后,除以恩诺沙星原形药物(M0)和N-去乙基代谢产物环丙沙星(M2)形式存在外,还有少量恩诺沙星脱羧代谢产物(M1)和恩诺沙星羟基化代谢产物(M3-1和M3-2)。该研究可为深入了解恩诺沙星在水生动物体内代谢产物及代谢机理提供理论基础,为在水产养殖生产中科学、合理地使用恩诺沙星提供参考。     

液相色谱-三重四级杆复合线性离子阱质谱法分析恩诺沙星在鲤鱼(Cyprinus carpio)中的代谢产物

采用液相色谱-三重四级杆复合线性离子阱质谱(HPLC/Q-TRAP MS),在正离子检测模式下,对鲤鱼(Cyprinus carpio)腹腔注射恩诺沙星给药后肝脏中的代谢产物进行分析.根据保留时间和各色谱峰质谱裂解规律,推测了恩诺沙星在鲤鱼肝脏中的代谢产物,同时,根据二级质谱碎片离子推测代谢途径及代谢产物结构.结果显示,恩诺沙星进入鲤鱼肝脏后,除以恩诺沙星原形药物(M0)和N-去乙基代谢产物环丙沙星(M2)形式存在外,还有少量恩诺沙星脱羧代谢产物(M1)和恩诺沙星羟基化代谢产物(M3-1和M3-2).该研究可为深入了解恩诺沙星在水生动物体内代谢产物及代谢机理提供理论基础,为在水产养殖生产中科学、合理地使用恩诺沙星提供参考.     

高效液相紫外法检测河鲀毒素

采用高效液相色谱法，用紫外检测器来检测TTX，有快速、简单和精确的优点。主要色谱条件为：Waters pico Tag^TM FAA色谱柱，0．01mol／L(NH4)3PO4溶液为流动相，紫外检测波长为205nm。该法在0．01～100μg／mL范围内，峰响应值与其浓度呈良好的线性关系，Y=11977x 11406，R^2=0．9997。最低检测限为6ng／mL。     

诺氟沙星对溶藻弧茵和恩诺沙星对迟缓爱德华菌的抗生素后效应

采用试管二倍稀释法测定诺氟沙星对溶藻弧菌s622、恩诺沙星对迟缓爱德华菌751的最小抑菌浓度(MIC)；采用菌落计数法测定诺氟沙星对溶藻弧菌、恩诺沙星对迟缓爱德华菌的抗生素后效应(PAE)。结果显示，诺氟沙星对溶藻弧菌s622在1／2MIC、1MIC、2MIC和4MIC浓度时的PAE分别为：0．47h、0．76h、1．26h和2．05h；恩诺沙星对迟缓爱德华菌751在1／2MIC、1MIC、2MIC和4MIC浓度时的PAE分别为：0．29h，0．48h，1．97h，2．30h。结果表明，诺氟沙星和恩诺沙星具有明显的PAE；该结果提示，在制定给药方案时可以适当延长给药间隔时间，减少给药次数，仍能维持抗菌效果。     

洛美沙星在鲤鱼体内的药物动力学及残留

用高效液相色谱 (HPLC)测定方法研究洛美沙星在鲤鱼 (Cyprinuscarpio)体内的药物动力学和残留。洛美沙星以 2 0mg/kg体重单剂量口灌鲤鱼后 ,测定其血浆药物浓度 ,最低检测限为 0 0 5 μg/mL ,动力学模型符合开放性有吸收二室模型 ,药物动力学参数 :表观分布容积 (Vd/F)、吸收半衰期 (tl/ 2kα)、分布半衰期 (t1/ 2α)以及消除半衰期 (t1/ 2 β)分别为 5 2 1L/kg、 0 31h、 1 0 2h、 2 0 0 3h ,其药时曲线下面积(AUC)为 110 92 μg·h/mL。以相同剂量单次口灌鲤鱼后 ,在不同的天数测定了鲤鱼皮肤、肌肉和肝胰脏中洛美沙星的残留 ,在给药 12天后 ,洛美沙星在各组织中均能检测到且残留均低于 0 0 8μg/g ,结果表明洛美沙星在鲤鱼体内消除缓慢、残留期较长 ,休药期不低于 12天。     

RP-HPLC同时测定中华绒螯蟹肝脏中诺氟沙星、环丙沙星和恩诺沙星残留

应用反相高效液相色谱法（RP—HPLC）建立测定中华绒螯蟹（Eriocheir sinensis）肝脏中诺氟沙星（NOR）、环丙沙星（CIP）和恩诺沙星（ENR）残留量的方法。考察了3种喹诺酮类药物残留的分离与流动相组成、pH值及乙腈含量的关系，优化了色谱条件。中华绒螯蟹肝脏成分复杂，脂肪类化合物含量高，色素等干扰物质多，极易干扰目标物的检测。通过改进前处理过程，采用振荡萃取-浓缩-高速离心萃取的前处理技术，有效减少了干扰物质对检测结果的影响。结果表明，该方法线性关系和重复性良好，样品中3种喹诺酮类药物残留的回收率在73．26％～85．82％，最低检测限为10μg／k。用该方法对肌肉注射给药后72h内中华绒螯蟹肝脏组织进行检测，其体内主要代谢物环丙沙星和恩诺沙星都可以检测到，说明本方法能够满足中华绒螯蟹体内诺氟沙星、环丙沙星和恩诺沙星残留量检测的技术要求。     

固相萃取-液相色谱串联质谱法同时测定养殖海水中17种喹诺酮类药物

建立了固相萃取-液相色谱串联质谱法(SPE-LC-MS/MS)同时检测养殖海水中17种喹诺酮类药物残留的分析方法。海水经酸化处理后，采用HLB固相萃取柱富集、净化目标化合物，通过对比水样在不同上样pH、淋洗液与洗脱液等条件下的回收率，以此对前处理方法进行优化。收集到的洗脱液经氮气吹干后用流动相定容至1 ml，待测。色谱流动相A相为0.1%甲酸水溶液，B相为乙腈，所有药物经梯度洗脱进行分离，在LC-MS/MS正离子模式的多反应监测模式下进行定性定量分析。17种化合物可以在10 min中内得到较好的分离，线性范围为1–200 ng/ml，线性相关系数均大于0.999，检出限均在2–10 ng/L范围内，定量限均在5–20 ng/L范围内。以空白海水为基质，在20、100、200 ng/L三个不同添加水平下采用内标法定量的加标回收率均在71.3%–125.0%范围内，相对标准偏差(RSD)为2.44%–12.27% (n=5)。采用该方法对黄海灵山湾近岸4个养殖场进行海水采集并检测，共检测出4种喹诺酮类药物，分别是恩诺沙星、氧氟沙星、诺氟沙星和环丙沙星，其中，恩诺沙星浓度最高。研究表明，该方法快速、可靠，适用于养殖海水中喹诺酮类药物的检测。     

水产品中噁喹酸残留量的检测技术研究

采用高效液相色谱荧光检测器研究了牙鲆、罗非鱼和对虾肌肉组织中噁喹酸残留的检测技术。样品前处理方法为：以无水乙酸乙酯作为提取液，样品经均质、离心后，将提取液旋转蒸发浓缩，用流动相溶解定容，正己烷除去脂肪。高效液相色谱条件为：色谱柱采用反相C18柱，荧光检测器激发波长为325nm，发射波长为369nm，流动相为0．02mol／L磷酸＋乙腈＋四氢呋喃（69：16：15），流速为0．8ml／min。噁喹酸标准曲线线性范围在10～200ng／ml。不同浓度水平下，噁喹酸的回收率为79％～93％，变异系数小于10％，检测限为10μg／kg。     

氟苯尼考对六种海洋致病弧菌的体外抗菌活性研究

测定了氟苯尼考等4种抗菌药对6种常见海洋致病弧菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。测试结果显示，氟苯尼考对6株弧菌的MIC值范围为2～4μg／mL，氯霉素为1～32μg／mL，土霉素为4～128μg／mL，恩诺沙星为0．125～1μg／mL；对多数弧菌氟苯尼考与氯霉素的抗菌活性相似。氟苯尼考对6株弧菌的MBC／MIC值范围为2-4，而氯霉素为2～16，表明氟苯尼考的杀菌效果优于氯霉素。对照药物恩诺沙星、土霉素的MBC／MIC值分别为1-2和2-8，恩诺沙星显示出优良的抗菌和杀菌活性。随着接种菌量增加，所测3种抗菌药的MIC随之升高；当接种量达到10^7ufu／mL时．药物对多数弧菌显示出微弱的作用。     

离子对高效液相色谱法测定日本囊对虾中喹诺酮类药物残留

采用离子对反相高效液相色谱法检测日本囊对虾中的喹诺酮类药物残留,使用Zorbax SB-C18柱(150mm×4.6mm,5μm)分离,流动相为乙腈∶水(0.01mol/L四丁基溴化铵溶液)=8∶92,流速为1.0ml/min,荧光检测器检测,激发波长为280nm,发射波长为480nm,柱温25℃。试验结果表明,每一种药物在一定的进样量范围内均有良好的线性关系,相关系数r均大于0.9998(n=6);氧氟沙星、环丙沙星和恩诺沙星的加标回收率均在80%以上,相对标准偏差小于10%。     

抑制性消减杂交技术在养殖鱼类免疫基因克隆中的应用

抑制性消减杂交技术(SSH)是近年来兴起的一种分离、克隆差异基因的新技术,它结合了消减杂交和抑制PCR的优点,具有操作简便、特异性强、背景低及重复性好的优点。目前已经用SSH技术鉴定出了鱼的许多免疫相关基因,如白细胞介素、趋化因子、肿瘤坏死因子、溶菌酶、NKEF、补体、干扰素及急性期蛋白基因等,对它们的结构和功能进行了较为深入的研究。本文对SSH技术在养殖鱼类中克隆的免疫基因进行了归纳与总结,旨为全面了解鱼类的抗病免疫基因提供基础资料。     

Dynamic properties of a flexible net sheet in waves and current—A numerical approach

The dynamic properties of a flexible net sheet exposed to waves and current were investigated by using a numerical model. The net was modelled by dividing it into super elements, and the structural and hydrodynamic forces were calculated for each element. The movement of the net was found from time integrating the equation of motion at each node. The model was used to study the influence of different parameters on the behaviour of the net. Through six different cases the impact of the following five parameters were investigated: (i) floater movement, (ii) wave period/height, (iii) current velocity, (iv) net solidity and (v) bottom weight.     

Dynamic properties of a flexible net sheet in waves and current—A numerical approach

The dynamic properties of a flexible net sheet exposed to waves and current were investigated by using a numerical model. The net was modelled by dividing it into super elements, and the structural and hydrodynamic forces were calculated for each element. The movement of the net was found from time integrating the equation of motion at each node. The model was used to study the influence of different parameters on the behaviour of the net. Through six different cases the impact of the following five parameters were investigated: (i) floater movement, (ii) wave period/height, (iii) current velocity, (iv) net solidity and (v) bottom weight.     

Evidence of homing of black rockfish Sebastes inermis using biotelemetry

ABSTRACT: The black rockfish Sebastes inermis is one of the most important fishery species along the coast from southern Hokkaido to Kyushu, Japan and is often found in rocks and Zostera areas. We conducted biotelemetry using coded ultrasonic transmitters to clarify the movement of the black rockfish that inhabited the seawall of the Kansai International Airport. We released 25 black rockfish at two points. One was the airport seawall and the other was side shallows off the Sensyu district. Seventeen black rockfish returned to their capture site after release. We used the V -test to determine whether the direction of movement was random or orientated. The black rockfish moved at random along the seawall within some hours after release ( P  > 0.05). Four hours after release, they moved to their home site intentionally ( P  < 0.0025).     

The quality of release habitat for reared juvenile flounder, Platichthys flesus, with respect to salinity and depth

Abstract  One prerequisite for a successful stocking programme is the choice of an appropriate release site, which would ensure good growth, survival and recruitment to the local fishery. The influence of different salinity regimes on habitat quality for juvenile flounder, Platichthys flesus (L.), was examined in Danish inshore waters using enclosures to study growth and survival. Three locations were chosen: Virksund (V) – constant low salinity at 10‰; Harre Vig (HV) – constant high salinity at 24‰; and Hjerk Nor – variable salinity, 0–25‰. Fish movement was examined, using a diffusion model, at the first two sites. At HV the fish had unrestricted dispersal opportunity resulting in average individual movement of 45 m day⁻¹. Fish at V were restricted in dispersal in one direction because of a dam, about which they would concentrate under certain hydrographical conditions. Growth was highest at the locations with stable salinity and mortality highest at the location with variable salinity.     

Hybrids as potential mediators spreading non-native genes: Comparison of survival,growth, and movement among native,introduced and their hybrid salmonids

Comparing multiple fitness components and potential movement of wild hybrids with their parental species is necessary to fully understand the consequences of human-mediated introgression, but studies tracking both parental species and their hybrids at the individual-level are limited. Here, we compared growth, survival and movement of sympatric introduced brook trout (BT: Salvelinus fontinalis) and native white-spotted charr (WSC: S. leucomaenis) with their hybrids (HYB) in a northern Japanese stream, using mark-recapture data (1,087 marked individuals) collected over 4 years (2013–2016). The mark-recapture data with a single cohort showed that HYB had a comparable or even higher growth rate to BT and WSC. In addition, there is no evidence that hybrid survival was lower than both parental species throughout the entire study period. Furthermore, HYB showed high mobility equivalent to WSC, while BT showed the lowest mobility. Although our previous studies have documented the reduction of BT distribution and lowered reproductive success of HYB, non-native genes can pose a threat to native WSC via relatively high survival, growth and/or mobility of HYB.     

Hatchery production of diploid and triploid clams, Tapes dorsatus (Lamarck 1818): a potential new species for aquaculture

To assess the potential for hatchery production of the venerid clam, Tapes dorsatus, diploid and triploid clams were produced and ongrown until the spat reached 5–8 mm in size. Triploidy percentages at metamorphosis that ranged from 56 to 85% were induced using a 15-min exposure to 1 mg/l cytochalasin B. No differences were observed in the growth rate of sibling diploid and triploid T. dorsatus larvae. Survival of diploid clams to pediveliger stage was however higher. Post-metamorphic growth and survival of diploid and triploid clams remained the same until the clams were removed from the hatchery to outdoor nurseries. Throughout the larval and early spat phases of production, T. dorsatus growth was similar to that reported for the Manila clam, Tapes philippinarum. Based upon its ease of culture, rapid growth and marketability, T. dorsatus are thought to have considerable aquaculture potential.     

Numerical modeling of chaotic behavior for small-scale movements of demersal fishes in coastal water

ABSTRACT:   The presented model involves the application of chaos theory to generate fish movements resulting from environmental stimuli. The model uses three steps within a model neural network such as input stimuli, central decision making and response output resulting in fish movements. The stimuli in the first step include the main abiotic and biotic factors, which could be quantified as an intensity parameter that was then normalized as a ratio between 0 and 1. The decision-making process can be generated using chaos dynamics with the stimuli parameters. The response of fish movements from the output signal representing movement speed and direction of fish can be re-regulated as main movement pattern depending on physiological state or life cycle by third response filtering. The simulation results seen as a movement pattern for sea bream and flounder using this neural chaotic model fitted very well to the observations of fish tracked in the sea by ultrasonic tracking methods. It was also revealed that the fish movement components generated as movement velocity and direction when in tidal flow had similar patterns to those patterns seen in field observations with similar irregular and chaotic variations with time.     

Disentangling individual movement between populations from effective dispersal in the facultative anadromous Salmo trutta L.

Gathering information on both individual movement and gene flow is rarely possible when studying dispersal among populations in fish species. It is, however, possible to assess both at a reasonable cost in Salmo trutta L. on the Atlantic coast of Europe where the facultative anadromous species is composed of discrete populations of brown trout residents occupying distinct river systems, but exchanging phenotypically distinguishable sea trout migrants. We performed two kinds of genetic analyses using individual microsatellite genotypes: the stock identification of sea trout entering each corridor and the estimates of effective dispersal through each corridor. We observed that individual movement (nonlocal individuals of each source population ranging from 4% to 35% of the sea trout run) never translates into effective dispersal except in one of four migratory corridors examined. The likely origin of this uniquely detected gene flow event is discussed in the light of well‐documented migratory fish management actions undertaken in the past in the studied area.     

Home range and seasonal movement of taimen,Hucho taimen,in Mongolia

Gilroy DJ, Jensen OP, Allen BC, Chandra S, Ganzorig B, Hogan Z, Maxted JT, Vander Zanden MJ. Home range and seasonal movement of taimen, Hucho taimen, in Mongolia.
Ecology of Freshwater Fish 2010: 19: 545–554. © 2010 John Wiley & Sons A/S Abstract –  Taimen, Hucho taimen, is the world’s largest salmonid and a prized sport fish. We used radio and acoustic telemetry to characterise movements of adult taimen in an extensive river system, the Eg–Uur, in north‐central Mongolia. Forty‐six taimen were tagged with transmitters (27 radio, 17 acoustic and 2 radio‐acoustic combined) and tracked from 2004 to 2008 using mobile surveys and fixed receivers. The mean home range of individual taimen tracked for an average of 2.4 years was 23 km (N = 41, range = 0.5–93.2 km). Of the fish with over 10 relocations (N = 16), 90% remained within a range of 38 km. Four distinct movement patterns were observed: (i) restricted core home range, (ii) core range with seasonal departures, (iii) core range with separate seasonal range and (iv) home range transfer. Movement was greatest in May and June (spawning and postspawning period) with another peak period of movement in September and October (water temperature cooling).