孔雀石绿及其代谢物无色孔雀石绿在鲫鱼肌肉中的代谢规律

研究了孔雀石绿及其主要代谢物无色孔雀石绿在鲫鱼肌肉组织内的残留消除趋势。对鲫鱼以200 ng/mL浓度孔雀石绿进行药浴1 h,取肌肉采用高效液相色谱法测定孔雀石绿及代谢物无色孔雀石绿的残留状况。结果表明,孔雀石绿代谢相对较快。代谢产物无色孔雀石绿残留时间较长,可以作为检测的标志物。     

孔雀石绿的代谢机理及生物毒性研究进展

孔雀石绿(Malachite green,MG)是1种三苯甲烷类工业染料,曾广泛用于治疗水产动物的水霉病、烂鳃病、寄生虫病等,但由于具有高毒、高残留、“三致”等毒性性质,许多国家禁止其在经济鱼类(观赏鱼除外)的养殖过程中使用,成为药物残留监控的主要内容之一。本文对孔雀石绿的化学性质、代谢规律及机理、生物毒性等研究现状进行了综述,旨在为药残的监控工作提供理论依据。     

液相色谱法测定鱼肌肉中孔雀石绿、结晶紫及其代谢物

孔雀石绿(MG)和结晶紫(GV)曾作为渔药使用,在鱼体内可分别代谢为无色孔雀石绿(LMG)及无色结晶紫(LGV),具有潜在的致癌性,已被许多国家禁用。本文研究了同时检测孔雀石绿、结晶紫及其代谢物无色孔雀石绿和无色结晶紫的液相色谱方法,建立了检测前处理方法及色谱条件。经过液液萃取和丙基磺酸阳离子树脂纯化,采用C18分离柱后串联二氧化铅柱氧化衍生的方法,同时检测母体药物及其代谢物,回收率>70%,重复性的相对标准偏差<15%,定量限分别为2μg/kg及1μg/kg。经样品实测,在一份鲈鱼样品中检测到无色孔雀石绿残留97μg/kg。     

甲基睾丸酮在尼罗罗非鱼(Oreochromis niloticus)肌肉中的代谢和消除规律研究

在实验水温(28±2)℃条件下,以60mg/kg单剂量甲基睾丸酮对尼罗罗非鱼混饲口灌给药后,采用超高效液相色谱-串联质谱联用(UPLC-MS/MS)法测定尼罗罗非鱼肌肉中的药物水平,研究甲基睾丸酮在尼罗罗非鱼肌肉中的代谢及消除规律。结果表明,肌肉中药物浓度时间数据符合一级吸收二室开放模型,主要药代动力学参数达峰时间T(peak)为3.04h,质量浓度C(max)为33.331ng/g,消除半衰期T(1/2)β为23.073h。肌肉中甲基睾丸酮浓度24h内呈指数型下降,具有较高的消除速率,其后消除得较缓慢,消除方程符合C=46.18e0.101t。29d后罗非鱼肌肉中甲基睾丸酮水平在美国FDA的检测限0.8ng/g以下,给药120d后肌肉中检测不到甲基睾丸酮残留。     

邻苯二甲酸二乙酯(DEP)在罗非鱼(Oreochromis niloticus × O.aureus)体内代谢动力学和残留研究

采用反相高效液相色谱.紫外检测法,测定了罗非鱼血浆、皮肤、肌肉、心肌和大脑中邻苯二甲酸二乙酯(DEP)含量,并研究了DEP在罗非鱼体内代谢动力学和残留特征.将DEP以20mg/kg灌服罗非鱼后,取样测定不同时间点血浆和组织中的DEP浓度,并利用3p97药物动力学软件并结合Excel 2003软件对实验数据进行处理和分析...     

黄芩苷在中国对虾体内的代谢及残留消除规律研究

在(24±1)℃下,中国对虾连续7 d给予100 mg/kg黄芩苷药饵,用高效液相色谱法(HPLC)测定对虾血液及其它组织中的黄芩苷含量,建立适合中国对虾体内黄芩苷含量的测定方法,研究黄芩苷在中国对虾不同组织中的残留消除规律.结果表明,HPLC法黄芩苷与杂质分离良好,4个浓度水平的黄芩苷在不同组织中的平均回收率在80.57%～96.25%之间,日内精密度在3.26%～5.85%之间,日间精密度在3.88%～5.73%之间.与其它给药方式相比,黄芩苷药饵给药吸收较慢,在血液、肝胰腺、肌肉、鳃中的达峰时间(Tmax)依次为6,3,6,3 h;黄芩苷在不同组织中的分布为肝胰腺>鳃>血液>肌肉.黄芩苷在肝胰腺、鳃、肌肉、血液中的消除半衰期(t1/2β)分别为29.62,27.50,25.20,23.73 h,消除速率从快到慢依次为血液>肌肉>鳃>肝胰腺;黄芩苷残留较少,在肌肉和血液中5 d已降到检测限以下,肝胰腺及鳃7 d降到检测限以下.本试验为含黄芩苷的药物的应用提供参考.     

磺胺甲噁唑在黑鲪肌肉中药代动力学及残留研究

在13℃条件下,采用单剂量灌服的方法给黑鲪100mg/kg体重的磺胺甲噁唑(SMZ),以HPLC法测定黑鲪肌肉中的药物浓度,用DAS2.0药物代谢动力学分析软件处理药时数据,结果表明:在13℃条件下,黑鲪单次灌服SMZ的药动学房室模型符合无时滞的二房室模型,分布半衰期t_(1/2α)为5.947h,消除半衰期t_(1/2β)为12.882h,达峰时间T_(max)为8h,峰值C_(max)为40.880μg/g.经过71d的代谢,肌肉中的药物含量下降到无公害食品中所规定的＜0.1mg/kg鱼体重.     

HPLC法研究磺胺类药物在水产动物体内的代谢和残留

目前国外获准用于防治水产动物疾病的药物有十几种,常使用的有磺胺嘧啶、磺胺－２,６－二甲氧嘧陡、磺胺异　唑、磺胺间甲氧嘧啶、磺胺噻唑、磺胺眯、甲氧苄氨嘧啶等。关于该类药物的药效学、代谢动力学及临床疗效方面的研究内容国内外已有不少报道,但国内在水产养殖方面的报道还不多,且主要偏重于药效和临床应用。随着世界经济一体化的发展,为保障食用者的健康,国际贸易对水产养殖产品安全卫生质量的要求越来越高,ＦＡＯ、欧美等要求对水产养殖全过程实行质量管理,提高产品质量卫生水平,我国也将全面落实以ＨＡＣＣＰ等为核心的科…     

海水实验围隔中放养罗非鱼的生态学效应

围隔实验和在围隔内进行的其他实验结果表明,台湾红罗非鱼(简称罗非鱼)对食物的选择性及其生态学效应与食物资源状况有关。放养罗非鱼的围隔中浮游植物生物量和生产力往往较高,但罗非鱼密度超过8000尾/hm2后可明显压制海洋原甲藻等大型鞭毛藻类。当罗非鱼食物丰富时其对围隔中桡足类的压制作用相对较弱,当其他食物资源缺乏时往往选择捕食个体较大的桡足类。放养罗非鱼对围隔水中氮浓度和COD的影响较小,但加快了磷的循环,罗非鱼密度超过8000尾/hm2的围隔中PO₄-P浓度明显高于对照围隔。     

抗菌药物在鱼、虾体内的残留及代谢动力学研究

随着人民生活水平的提高及水产品出口规模的扩大,我国水产养殖业发展非常迅速。但是在水产养殖中药物的滥用和非法使用问题严重,药物在水产品中大量积累,严重威胁消费者身体健康。伴随中国加入WTO。欧盟、美国和日本等国家对中国水产品中抗菌药物的残留提出了更高要求,中国水产品出口在短时间内多次遭遇“绿色壁垒”。自2002年1月25日欧盟宣布对中国出口的动物源性食品实行禁运后,中国水产品出口受到了极大的损失。对水产品中抗菌药物的残留研究已经迫在眉睫,国外学者开展这方面的研究较早,也已经取得了一定的进展．国内这方面的研究较少。本文就水产养殖中各类常用的抗菌药物在鱼、虾体内的残留研究现状、药物代谢动力学及检测方法等进行综述,以期能够对中国水产养殖中药物的应用及相关研究起促进作用。     

Degradation of malachite green dye by Tenacibaculum sp. HMG1isolated from Pacific deep-sea sediments

A deep-sea bacterium from the Pacific Ocean identified as Tenacibaculum sp. HMG1 was found to have strong malachite green(MG) degradation activity. The MG tolerance and decolorizing activities of strain HMG1 were confirmed by bacterial growth and high-performance liquid chromatography(HPLC) analyses. Strain HMG1 was capable of removing 98.8% of the MG in cultures within 12 h and was able to grow vigorously at 20 mg/L MG. A peroxidase gene detected in the genome of strain HMG1 was found to be involved in the MG biodegradation process. The corresponding recombinant peroxidase(r POD) demonstrated high degradative activity at 1 000 mg/L MG. Based on the common candidate intermediates, strain HMG1 was inferred to have one primary MG degradation pathway containing r POD. In addition, five other candidate intermediates of the r POD-MG degradative process were detected. The optimal conditions for MG degradation were determined and showed that strain HMG1 and the r POD enzyme could maintain high bioactivity at a low temperature(20°C), variable p H values(6.0–9.0), higher salinities(100 mmol/L) and other factors, such as multiple metal ions, H_2O_2 and EDTA.MG-tolerant strain Tenacibaculum sp. HMG1 and its peroxidase have prospective applications as environmental amendments for MG degradation during coastal remediation.     

EDTA对罗非鱼嗅觉的影响及其解毒作用

将５种浓度的ＥＤＴＡ和９种含有不同比例的重金属离子、ＥＤＴＡ混合液分别灌注罗非鱼嗅囊,观测这些溶液中甲硫氨酸引起的嗅电图的影响,研究ＥＤＴＡ对重金属染毒钱瓣可行性和最佳配方,并探讨了其可能机理,结果如下：（１）ＥＤＴＡ对ＥＯＧ有抑制和促进两方面影响。当浓度低于３７２ｍｇ／ｄｍ＾３时,ＥＤＴＡ抑制ＥＯＧ反应、浓度越低,抑制作用越明显。当浓度大于３７２ｍｇ／ｄｍ＾３时,则促进ＥＯＧ反应,浓度越高,促进     

太平洋深海沉积物来源的菌株Tenacibaculum sp.HMG1对孔雀石绿降解特性的研究

本文在太平洋深海沉积物中分离得到一株孔雀石绿降解菌株,鉴定命名为Tenacibaculum sp.HMG1。通过菌株生长实验和高效液相色谱的研究表明, HMG1菌株可以在20 mg/L的孔雀石绿中维持较快的生长速率,并且在12 h内可降解98.8%的孔雀石绿,这证明该菌株具有很高的孔雀石绿耐受能力和降解活性。通过基因组测序在HMG1菌株发现一条过氧化物酶基因可能参与了孔雀石绿的降解,随后利用原核表达获得了相应的重组蛋白。实验表明,该重组过氧化物酶具有极强的活性,可在1000 mg/L的孔雀石绿中发挥降解功能。本文利用液相色谱-质谱联用（LC-MS）技术对孔雀石绿的菌株降解产物和重组酶降解产物进行鉴定,并基于鉴定结果推测了两种降解途径。结果发现两种降解方式存在共同的降解途径。此外,孔雀石绿降解条件的实验结果证明重组过氧化物酶可以在低温（20℃）、复杂的pH值（6.0–9.0）、高盐度（100 mmol/L）、金属离子和EDTA等反应条件下依旧维持很高的孔雀石绿降解活性。以上实验结果表明,HMG1菌株和重组过氧化物酶均在孔雀石绿污染生物修复方面具有很大潜力。     

Grazing effects by Nereis diversicolor on development and growth of green algal mats

Nereis diversicolor is generally considered to be a predator and deposit feeder, but have also been found to graze on benthic algae in shallow coastal areas. In this study we investigated the grazing effects on the development and growth of green algae, Ulva spp. Algal growth was studied in an experiment including two levels of sediment thickness; 100 mm sediment including macrofauna and 5 mm sediment without macrofauna, and three treatments of varying algal biomass; sediment with propagules, sediment with low algal biomass (120 g dry weight (dwt) m^− 2) and sediment with high algal biomass (240 g dwt m^− 2). In the 100 mm sediment, with a natural population of macrofauna, N. diversicolor was the dominating (60% of total biomass) species. After three weeks of experimentation the result showed that N. diversicolor was able to prevent initial algal growth, affect growth capacity and also partly reduce full-grown algal mats. The weight of N. diversicolor was significantly higher for polychaetes in treatments with algae added compared to non-algal treatments. There were also indications that a rich nutrient supply per algae biomass counteracted the grazing capacity of N. diversicolor.     

O,p'-DDT induction of vitellogenesis and its inhibition by tamoxifen in Nile tilapia (Oreochromis niloticus)

In order to investigate the mechanism by which o,p'-DDT disrupts endocrine functioning of Nile tilapia in vivo, the estrogenicity of o,p'-DDT was investigated in conjunction with 17beta-estradiol (E2) and tamoxifen. Mature, male tilapia were treated intraperitoneally with o,p'-DDT (60 mg/kg, one dose) or E2 (5 mg/kg, four doses) in the presence or absence of tamoxifen (5 mg/kg, six doses) for 12 days and then plasma vitellogenin (Vtg) (measured as alkaline-labile phosphorous), E2, and testosterone (T) were measured. Vtg levels were increased dramatically by E2 (1,744 +/- 171 microg/ml) and moderately by o,p'-DDT (82 +/- 15 microg/ml) compared with controls (23 +/- 3.5 microg/ml). Tamoxifen alone had no effect on Vtg production, but inhibited both E2 and o,p'-DDT stimulated vitellogenesis. T levels were reduced with E2 administration (1,688 +/- 383 pg/ml) and declined further with the combined treatment of E2 and tamoxifen (281 +/- 70 pg/ml), compared with controls (6,558 +/- 1,438 pg/ml). Tamoxifen or o,p'-DDT alone did not affect T levels, but their combined treatment did (2,069 +/- 647 pg/ml). The results of this study suggest that o,p'-DDT is weakly estrogenic in male tilapia, and that this activity may be mediated through the estrogen receptor.     

西太平洋帕劳砗磲高分辨率氧同位素记录及其指示的气候环境变化

砗磲是海洋中最大的双壳类贝壳,其碳酸盐壳体通常具有年纹层和天纹层,是一种理想的高分辨率古气候研究载体.氧同位素是砗磲古气候研究中最常用的指标之一,但在将其应用于古气候重建之前,通常需要对其现代地球化学过程进行准确的校准.帕劳群岛位于西太平洋暖池西北边缘,其珊瑚礁盘具有丰富的砗磲壳体资源,为开展古气候研究提供了丰富的材料...     

荧光淬灭剂TrueBlack Lipofuscin对珊瑚自体荧光的消除及其在免疫荧光标记中的应用

自体荧光是生物体内部组织成分在吸收光时自然发出的荧光,会影响人工标记荧光的有效识别,严重干扰对抗体标记的目的蛋白荧光的观察及数据分析,在珊瑚冰冻切片免疫荧光标记研究中尤为突显。为更有效地去除珊瑚组织切片中的自体荧光,本研究以共生模式种花伞软珊瑚（Xenia sp.）为研究对象,首次尝试在花伞软珊瑚冰冻切片免疫荧光染色中,使用一种在小鼠和人体组织切片中常用的TrueBlack Lipofuscin自体荧光淬灭剂,并与已有研究中的酒精梯度脱水法进行比较。结果表明,使用荧光淬灭剂TrueBlack Lipofuscin后,花伞软珊瑚组织切片中的自体荧光强度是传统的酒精梯度脱水法的52.4%,且在Cy5通道下效果最佳。同时,通过珊瑚免疫荧光标记实验表明使用TrueBlack Lipofuscin的实验组特异性荧光信号强度为对照组的205.3%,背景荧光和自体荧光基本去除。     

Seasonal Blooms of the HAB Dinoflagellate Alexandrium taylori Balech in a New Mediterranean Area (Vulcano, Aeolian Islands)

Abstract. High-biomass blooms of A. taylori Balech have recently been spreading over new Mediterranean areas, with evident adverse effects on the marine ecosystem. In 1999–2000, a new Mediterranean locality was affected by blooms of A. taylori: the West Bay of Vulcano (Aeolian Islands, Tyrrhenian Sea), with maximum cell densities of 1.2 × 10⁷ cells 1⁻¹ in August 1999 and 4.0 × 10⁶ cells 1⁻¹ in August 2000 observed together with yellowish water discoloration. The seawater samples contained high concentrations of nutrients as DIN (Dissolved Inorganic Nitrogen), especially NH₃-N with values of 14.4 μM and TOT-P (Total Phosphorus) with values of 3.2 μM due to the anthropic presence and discharge of untreated sewage. The climatic conditions also seem to influence the occurrence and spreading of the A. taylori blooms in the Vulcano Bay. Clonal cultures of A. taylori , established from Italian and Spanish seawater samples, were used for the sequence analyses of the 5.8S rDNA gene and ITS regions in order to study the genetic variability of different geographical populations of Alexandrium species in the Mediterranean area and to further develop the molecular markers for HAB key-species.     

罗非鱼、南美白对虾对重金属富集的研究

对广东省养殖的罗非鱼Oreochromis niloticus及南美白对虾Litopeneaus vannamei体内Pb、Cd、Hg、As 4种重金属富集及分布状况进行研究。比较了不同规格罗非鱼及南美白对虾体内重金属差异。分析重金属在罗非鱼及南美白对虾不同部位的分布状况。结果表明,罗非鱼及南美白对虾体内Pb、Cd、Hg、As均有不同程度的检出,但含量均低于国家限定标准(NY5125-2002);其中Pb的含量最高,罗非鱼和南美白对虾体内Pb含量最高值分别为限量的39.32%和42.18%;重金属含量因其规格大小及部位而异,趋势为大规格样品重金属含量相对较高,重金属含量在罗非鱼及南美白对虾体内的增长情况与其体重具有正相关性;罗非鱼内脏重金属含量明显高于肌肉组织,虾头为虾重金属富集的主要部位。     

小角毛藻对球形棕囊藻囊体形成的影响

球形棕囊藻具有异型的生活史,能够在单细胞和囊体两种不同形态之间转换。球形棕囊藻藻华现场发现,囊体有时附着在角毛藻上。为研究角毛藻对囊体形成的影响和机制,进行球形棕囊藻和小角毛藻的共培养实验,观察球形棕囊藻囊体的形成和生长状况,研究小角毛藻对球形棕囊藻囊体形成的影响。结果证明:混合培养下,小角毛藻和球形棕囊藻的囊体附着在一起,在较短的生存周期内,囊体的密度明显高于单独培养时的密度;而单独培养体系下,棕囊藻单细胞密度和囊体体积均高于混合培养,囊体的存在时间较长。小角毛藻的存在对囊体的形成具有促进效应,这对于球形棕囊藻囊体具有重要的生态意义。硅藻的存在更能体现球形棕囊藻竞争优势。