两种喂食方式下微囊藻毒素-LR在罗非鱼肝脏和肌肉中动态变化的比较(Dynamical Response of Microcystins-LR in Liver and Muscle of Tilapia（Oreochromis niloticus）in Two Feeding Conditions of Cyanobacteria)

试验以尼罗罗非鱼作为受试动物,通过直接投喂未经处理的新鲜藻细胞和喂食相同量经超声波破碎过的藻细胞,比较了罗非鱼在摄食完整蓝藻细胞与摄食破碎的蓝藻细胞时微囊藻毒素-LR（MC-LR）在其体内的动态变化规律及代谢差异. 结果表明,无论是喂食破碎蓝藻细胞还是未破碎的蓝藻细胞均可在罗非鱼肝脏和肌肉中检出MC-LR,细胞破碎组肝脏和肌肉的富集能力高于未破碎组. 富集第3d,即可在肝脏和肌肉中检出不同程度的MC-LR,肝脏中MC-LR的含量显著高于肌肉,随着富集时间的延长,MC-LR的含量总体呈上升趋势,破碎组肝脏和肌肉分别在染毒的16d和11d达到最大值（分别为2.021μg·g-1和0.071μg·g-1）;未破碎组则分别在染毒的24d和21d达到最大值（分别为1.856μg·g-1和0.036μg·g-1）. 整个富集阶段破碎组肝脏MC-LR的平均值为1.171μg·g-1,略高于未破碎组的1.029μg·g-1;破碎组肌肉含量的平均值0.051μg·g-1,高于未破碎组的0.029μg·g-1. 将细胞未破碎组的罗非鱼进行释放阶段的试验. 结果表明,罗非鱼的肝脏能快速清除MC-LR,释放13d时达到释放阶段的最低值（为0.241μg·g-1）,与肝脏相比,肌肉对毒素的清除要缓慢得多.     

一株微囊藻毒素-LR降解菌的降解特性

研究了从某人工湖底泥中筛选出的1株能够降解微囊藻毒素-LR(MC-LR)的菌株JM13的降解特性.实验结果表明,当加入50 mg·L-1淀粉作为外加碳源时,菌株对初始浓度为0.5 mg·L-1MC-LR的降解率可达44.5%;低浓度重金属Cu2+的加入可在一定程度上提高菌株对MC-LR的降解,降解率可达到44.8%;随着时间的延长,菌株对MC-LR的降解率不断增大,到第10天达到52.6%,当投入菌龄为36 h的菌悬液时,降解率可达到55.4%.对该菌株细胞表面疏水性(CSH)的测试结果显示,在菌JM13细胞表面疏水性最大的情况下(即有机∶相水相=3∶4时),添加淀粉及低浓度(0.5 mg·L-1)Cu2+可以在一定程度上抵御降解过程中细菌表面疏水性的降低,使菌体细胞能更好地同污染物接触,从而在一定程度上提高菌株对MC-LR的降解.     

微囊藻毒素-LR通过线粒体途径诱导人支气管上皮细胞凋亡

探讨线粒体Caspase依赖性途径是否参与微囊藻毒素-LR(microcystin-LR,MC-LR)诱导人支气管上皮细胞(human bronchial epithelial cells,16HBE)凋亡过程。将处于对数生长期的16HBE分别暴露于终浓度为0(对照组)、2.5、5、10μg·m L-1的微囊藻毒素-LR和10μg·m L-1MC-LR+50μmol·L-1Caspase广谱抑制剂Z-VAD-FMK,持续24 h和48 h。检测细胞凋亡率,线粒体跨膜电位(ΔΨm),Caspase-3和Caspase-9相对表达量。结果显示,与对照组相比,各浓度染毒组细胞凋亡率和Caspase-3、Caspase-9相对表达量均升高,10μg·m L-1MC-LR染毒组线粒体膜电位降低;与10μg·m L-1MC-LR组相比,10μg·m L-1MCLR+50μmol·L-1Z-VAD-FMK组细胞凋亡率明显降低,Caspase-3和Caspase-9相对表达量降低,差异均有统计学意义(P0.05)。且随着MC-LR染毒浓度的升高或染毒时间的延长,16HBE细胞凋亡率和Caspase-3、Caspase-9相对表达量呈升高趋势。研究表明,MC-LR可以通过线粒体Caspase依赖性途径诱导16HBE细胞凋亡。     

叶绿素作用下微囊藻毒素-LR的光降解

研究了蓝藻中含量最多的色素--叶绿索在微囊藻毒素-LR(MC-LR)光催化降解中的作用,考察了波长和光强等对叶绿素催化的MC-LR光降解的影响.结果表明,叶绿素能导致MC-LR在日光照射下光降解,而且MC-LR的降解与其浓度呈正相关.在叶绿素浓度为0.1mg·ml-1的条件下,日光照射30 minMC-LR的去除率即可达到95%以上,而叶绿素浓度为0.05 mg·ml-1和0.01 mg·ml-1时,照射30 min后MC-LR的去除率分别为65%和56%.日光中紫外区(200-300 nm)的光,在光催化降解MC-LR中起主要作用,与体系最大吸收光谱范围一致的240 nm的光激发效果最好,光催化降解作用也最强.光照强度是影响MC-LR降解的重要因素,光强越大,MC-LR的降解率和降解速度越大.     

微囊藻毒素-LR对罗非鱼鳃组织活性氧自由基含量及相关抗氧化酶活性的影响

应用腹腔注射的方式,研究了微囊藻毒素MC-LR对罗非鱼（Oreochromis niloticus）鳃组织活性氧自由基含量（ROS）及过氧化氢酶（CAT）、超氧化物歧化酶（SOD）活性的影响。结果表明：在MC-LR的胁迫下,罗非鱼鳃组织产生了大量的ROS,并引起了SOD和CAT 2种抗氧化酶活性的变化。SOD和CAT在清除过量的ROS时表现出的动态变化过程不同,SOD的变化曲线是多阶段的,而CAT则表现为先诱导后恢复的趋势。SOD和CAT在变化趋势上有所不同,但均表现为明显的时间与效应的正相关关系和剂量与效应的正相关关系。这也为将鳃组织中SOD和CAT作为抗氧化生物标志物来监测微囊藻毒素对罗非鱼胁迫作用提供了一定的参考。试验为研究微囊藻毒素对罗非鱼的生态毒理学作用提供相应的资料,并为罗非鱼质量安全问题的研究打下一定的基础。     

微囊藻毒素-LR对鸡肝脏的氧化损伤影响

蓝藻虽然能产生有毒的生物毒素,但是也含有较高的蛋白质。为探索蓝藻饲料化利用的可能性,本文通过腹腔注射的方式研究了微囊藻毒素-LR(MC-LR)对崇仁麻鸡的半数致死剂量(LD_(50))及其对肝脏的氧化损伤。实验设计了4个剂量组(对照组、5、10和20μg·kg~(-1)MC-LR),并在1、3、12、24和48 h检测了谷胱甘肽(GSH)含量以及谷胱甘肽S-转移酶(GST)、谷胱甘肽过氧化物酶(GPX)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性变化。结果表明,MC-LR对崇仁麻鸡的LD_(50)值为34.67μg·kg~(-1)体重(bw),95%的置信限为33.51～35.83μg·kg~(-1)bw。在MC-LR的作用下,鸡肝出现了氧化应激现象。3个染毒组鸡肝中GSH含量呈现先下降而后上升恢复至正常水平的趋势,GST酶活力表现为先上升而后下降至正常值的趋势,这说明GSH和GST共同参与了鸡肝中MC-LR的解毒;鸡肝中GPX酶活性在前3小时先保持不变(除了1 h的高剂量组),随即显著上升,这说明GPX和GSH共同参与了鸡肝中过量活性氧自由基(ROS)的清除,GPX可以作为监测MC-LR引起鸡毒性作用的生物标志物。CAT酶活力表现为先显著下降(P<0.05)而后快速上升至正常值的趋势,SOD酶在整个实验期间几乎保持稳定,这可能与SOD酶活性较高所致。     

微囊藻毒素-LR致小鼠肝细胞DNA-蛋白质交联作用的研究

为探讨微囊藻毒素-LR致小鼠肝细胞的DNA-蛋白质交联作用,将20只昆明雄性小鼠随机分为4组:1个对照组和3个染毒组,采用腹腔注射进行染毒7d,染毒剂量分别为3.0、6.0和12.0μg·kg-1,检测小鼠肝细胞DNA-蛋白质交联程度.结果显示,3.0、6.0和12.0μg·kg-1微囊藻毒素-LR均可导致小鼠肝细胞显著的DNA-蛋白质交联作用(与对照组相比,p<0.01,p<0.01,p<0.05),当微囊藻毒素为6.0μg·kg-1时,这种作用最明显.     

微囊藻毒素在鲋鱼体内生物富集及其体内的抗氧化反应

为了解微囊藻毒索在鲋鱼Carassius auratus L.体内生物富集作用,用LC/MS监测不同时间的鲋鱼肝脏、肌肉,以及饲养用水中痕量的微囊藻毒素.结果显示,肌肉组织中MC-RR和MC-LR的含量在18 d时达到峰值,分别为7.87 ng·g~(-1)和2.18 ng·g~(-1);而肝脏组织中MC-RR和MC-LR的含量在鲋鱼暴露9天时达到最高值,分别为25.30 ng·g~(-1)和33.27ng·g~(-1).研究结果支持肝脏组织是MCs的主要靶向器官,并且表明肝脏组织对MC-LR的富集量远大于MC-RR,而肌肉组织更易于积累MC-RR.文章还研究了鲋鱼体内的抗氧化酶(SOD、CAT、GST、GR酶)的活性变化,对MCs介导的氧化胁迫进行了评估.通过分别测定暴露不同时间点(3、9、18 d)肝脏和肌肉组织中的抗氧化酶的活性,发现它们的活性与组织中MCs的含量基本呈正相关,可能对MCs介导的氧化胁迫有缓解作用.以上表明,MCs能在鱼体内积累,抗氧化系统虽可缓解,但不能完全降解.因此食用被MCs污染的鱼类存在潜在的食品安全风险.     

微囊藻毒素-LR对雄性黑斑蛙精巢中CYP46A1,CYP2H2和CYP2G1 mRNA表达的影响

微囊藻毒素-LR(microcystin-LR,MC-LR)是分布最广泛和毒性最强的一种微囊藻毒素,对水生动物造成潜在的健康威胁。大量科学研究证实,动物体中的细胞色素P450酶(CYP)参与内源性物质及外源毒性物质的代谢过程。为探究两栖动物生殖器官中的CYP酶对低剂量MC-LR生殖毒效应的调节作用,选择雄性黑斑蛙(Rana nigromaculata)为受试动物,采用静态置换法和体内暴露方法,分别暴露于0、0.1、1和10μg·L~(-1)MC-LR溶液0、7和14 d;用实时荧光定量PCR方法检测精巢中CYP46A1、CYP2H2和CYP2G1的mRNA表达水平。结果表明,暴露于0.1、1和10μg·L~(-1)MC-LR 14 d后,CYP46A1在mRNA水平分别上调了1.86、1.65和1.22倍,CYP2H2在mRNA水平分别上调了4.62、1.80和1.04倍,CYP2G1的mRNA水平分别上调了2.63、2.16和1.56倍。MC-LR在1μg·L~(-1)剂量下暴露7 d后,CYP46A1、CYP2H2和CYP2G1 mRNA水平均出现显著上调。上述研究表明,微囊藻毒素对黑斑蛙精巢3种CYP基因在mRNA水平上都存在低剂量刺激效应。低剂量MC-LR能诱导黑斑蛙精巢中CYP46A1转录水平变化,促进胆固醇转化为24S-羟化胆固醇,潜在破坏雄性黑斑蛙精巢中胆固醇水平的平衡; MC-LR也能够诱导精巢中CYP2H2和CYP2G1转录水平的变化,潜在调节CYP2H2和CYP2G1转录水平,进而影响MC-LR的代谢作用。     

微囊藻毒素对几种淡水微藻的生长和光合活性的影响

微囊藻毒素(MC)是富营养化淡水水体中最常见的藻类毒素,而MC对藻类生长效应的影响却鲜见报道.通过模拟培养实验,研究了不同质量浓度的MC-RR对淡水藻类的生长和光合效能的影响.结果显示,100 μg·L-1以下的MC-RR对产毒铜绿微囊藻Ds(Microcystis aeruginosa Ds)作用并不明显;相反,100 μg·L-1 MC-RR对铜绿微囊藻无毒株854(Microcystis aeruginosa 854)有显著的杀藻效应,表明MC可能改变浮游植物种群中产毒与非产毒微囊藻的比例.MC-RR对其它藻类的作用因种类不同而效果各异.100 μg·L-1 MC-RR可显著抑制细长聚球藻(Synechococcus elongatus)的生长,并诱使水华束丝藻(Aphanizomenon flos-aquae)发生溶藻效应;100 mg.L-1显著降低了聚球藻和束丝藻的光合活性,表明微囊藻毒素对藻类生长的抑制与其对光合活性的抑制有关.100 μg·L-1以下的 MC-RR对蛋白核小球藻(Chlorella pyrenoidosa)、斜生栅藻(Scenedesmus obliquus)、水华鱼腥藻(Anabaena flos-aquae)的生长没有明显影响;1 000 μg·L-1 MC-RR 则可促进这三种藻的生长,但对小球藻和鱼腥藻的光合效能没有明显影响.以上结果说明MC参与浮游植物的种间相互作用与种群调节.     

亚硝态氮胁迫下罗非鱼对海豚链球菌的易感性及血清超氧化物歧化酶的响应

为探索亚硝态氮(NO-2-N)胁迫与罗非鱼(Oreochromis niloticus)海豚链球菌(Streptococcus iniae)病爆发的关系,揭示罗非鱼海豚链球菌病爆发的环境机制,研究了不同质量浓度NO-2-N(<0.01、0.75、1.50、3.00和5.00mg.L-1)胁迫下罗非鱼对海豚链球菌的易感性及罗非鱼血清超氧化物歧化酶(SOD)活性的响应。结果表明,感染海豚链球菌的罗非鱼死亡率随NO-2-N浓度的升高而升高,并与暴露时间呈显著正相关(P<0.05),在暴露12～13 d时死亡率达到稳定状态,<0.01 mg.L-1浓度组在暴露13 d时,0.75、1.50、3.00和5.00 mg.L-1浓度组在暴露12 d时累积死亡率分别为20.00%、23.33%、30.00%、43.33%和46.67%。罗非鱼血清SOD活性随NO-2-N浓度的升高而降低,试验期间≤1.50 mg.L-1NO-2-N处理组血清SOD活性与对照组相比差异均始终未达显著水平(P>0.05),试验48、96和120 h时≥3.00 mg.L-1NO-2-N处理组血清SOD活性显著低于对照组(P<0.05)。认为高质量浓度(≥3.00 mg.L-1)NO-2-N能够显著降低罗非鱼的免疫力,增加其对海豚链球菌的易感性。     

Effects of daily nickel intake on the bio-accumulation,body weight and length in tilapia (Oreochromis niloticus)

Nickel pollution is a serious environmental problem, and its effects may provoke alterations in the ecosystem and in organism of animals and humans. Dermatitis, eczema, and asthma are some illnesses caused by Ni²⁺ poisoning. In this work, fish fed either Ni²⁺-enriched pellets or commercial pellets were studied. The amount of Ni²⁺ in fish were measurements by adsorptive stripping voltammetry (AdSV) with dimethylglyoxime as a complexing agent. The analysis of Ni²⁺ in fish by AdSV established that its accumulation occurs principally in viscera (670.86 ± 5.82 µg g^?1), in the head (697.12 ± 2.77 µg g^?1) and in the muscle (405.82 ± 3.26 µg g^?1), both after 12 months of experiments. Ni²⁺ adsorbs preferentially in organs such as the stomach, the intestine, and the kidneys and acts in the central nervous system as well. Tilapia growth and mass were significantly affected by Ni²⁺ poisoning. From statistical analysis, observed that the results for lengths, weights and metal concentration were different for each sampling at significance level of p < 0.05. The Ni²⁺ concentration in tilapia was enough to cause the death of tilapias; however, it did not occur because the presence of Zn²⁺ might act as protective agent of heavy metals.     

不同价态无机砷对罗非鱼肝脏砷积累及GSH/GST解毒代谢的影响

低剂量中长期暴露下的氧化胁迫是砷对水生生物致毒的重要机制之一。本文通过对罗非鱼进行32 d的食物相砷暴露,测定不同时间点罗非鱼肝脏中谷胱甘肽(glutathione,GSH)含量和谷胱甘肽巯基转移酶(glutathione S-transferase,GST)活性,揭示不同价态无机砷对罗非鱼肝脏中GSH/GST的影响机制。经三价砷(As(III))暴露后,砷含量在2 d内显著增加而在随后的30 d内无显著性差异; 0～2 d内GSH含量显著增加,后降低,13 d后GSH含量均低于空白组; 0～6 d GST活性均大于空白组,6～8 d GST活性降低,8 d后活性高于空白组,且32 d达到最大值。经五价砷(As(V))暴露后,罗非鱼肝脏中砷含量逐渐增加,在20 d时达到最大值而后无显著性差异; 0～2 d时GSH含量降低,随后逐渐增加,在16 d达到最大值,16 d后GSH含量均低于空白组; 0～8 d时GST被大量诱导合成,8～20 d时GST合成被抑制,20 d后活性增加,在32 d达到最大值。As(III)和As(V)对罗非鱼GSH/GST的不同影响与其在罗非鱼体内的积累量有关。As(III)暴露后各时间点罗非鱼肝脏中的砷含量与GSH含量呈统计学正相关,而As(V)暴露无明显相关性。这是因为As(V)进入罗非鱼肝脏后会还原为As(III),进而GSH作为可提供巯基的还原剂而被大量消耗。另外,As(III)暴露后各时间点罗非鱼肝脏中的砷含量与GST活性呈显著负相关,而As(V)暴露却呈现出很强的滞后性,这是由于进入生物体内的As(V)需转化为As(III)后,才可直接作用于酶系统。可见,不同形态砷对水生生物的致毒机制需进一步深入研究。     

Protective action of zinc against lead poisoning in tilapia Oreochromis niloticus

The accumulation of lead (Pb) in tilapias Oreochromis niloticus, acclimatized in cement tanks, and fed once a day with pellets of commercial food with lead nitrate, was investigated by square wave anodic stripping voltammetry (SWV). The head (including head bones, proteins, and brain) was the main target of lead uptake by tilapias, with a small amount in muscle tissue. A high concentration of Pb was found in feces which indicated low absorption by the organism when fed with high Pb concentration. The protective effects of zinc (Zn) against the Pb poisoning, the accumulation of Pb in the heads of the fishes of 107.8?±?3.1?mg?kg^?1 and in the viscera of 57.1?±?2.9?mg?kg^?1 with consequent decrease of calcium (Ca) concentration, were observed. In spite of the experimental time of 16 months and higher Pb concentration in the dietary food of 320?mg?kg^?1, no death of tilapias was observed due to the protective action of Zn.     

贫营养型水库中罗非鱼对浮游植物的影响:围隔实验

于2009年9～11月在贫营养型的大型深水水库(流溪河水库)中进行了围隔(体积约为85 m3)实验,分析了罗非鱼对浮游植物的影响.根据水库中现有鱼类的生物量,设置加鱼处理组(2 g/m3)和无鱼对照组两组.每组有3个重复,实验开始后每周采样一次,测定水质与采集浮游生物定性和定量样品.实验结果表明,加鱼处理组和对照组的总氮、可溶性无机氮没有明显差别,实验后期的总磷与正磷有一定差别,处理组略高于对照组.加鱼处理组浮游植物丰度及生物量、叶绿素a的含量显著高于对照组(P<0.05),且透明度低于对照组,个体小于30μm的浮游植物种类的丰度与生物量较明显增加,浮游动物丰度显著低于对照组(P<0.05),但是生物量差异不显著(P>0.05).总的来说,在贫营养条件下,低生物量的罗非鱼对水库浮游植物群落结构仍有着明显的影响,罗非鱼排泄产生的上行效应大于摄食产生的下行效应.     

Observations on metal concentrations in commercial landings of two species of tilapia (Oreochromis mossambicus and Oreochromis niloticus) from reservoirs in six river basins in Sri Lanka

Samples of the muscle of two species of tilapia (Oreochromis mossambicus and O. niloticus; 17–20 cm length) were obtained from at least one reservoir in each of the six river basins (Aruvi Aru, Kala Oya, Kirindi Oya, Ma Oya, Mahaweli, and Walawe Ganga catchments) in Sri Lanka. The metals Ca, Cu, Fe, K, Mg, Mn, Na, and Zn were consistently detected in the muscle tissue. Overall, there were few differences in the concentration of metals between the two species of fish, although there were also some statistically significant differences (p < 0.05) in the concentrations of some metals in fish obtained from some of the reservoirs. Aruvi Aru stands out as a river basin in which the two fish species have significantly lower concentration of metals when compared to other river basins. The concentration of the metals studied were below WHO and FSANZ guideline values for fish, suggesting that the consumption of the metals found in tilapia from these reservoirs poses little risk to human health.     

Toxic effects of dietary of Al3+ ions in tilapias (Oreochromis niloticus) and protective effect of Zn2+ ion

The lethal effects of aluminum ion (Al³⁺) in tilapia (Oreochromis niloticus) raised in concrete tanks were investigated. Tilapias were fed daily with commercial feed enriched with known concentrations of Al³⁺ and analyzed by differential pulse anodic stripping voltammetry (DPASV). The concentrations of Al³⁺ in feces, water, muscle tissue, viscera, and heads were determined every 3 months for a period of 365 days. The Tilapia head was the most affected tissue by Al³⁺. In general, Al³⁺ bioaccumulation reached the lethal dose (LD₅₀) after 335 days of experiment as follows: 34.9?mg?kg^?1 (muscle tissue), 88.2?mg?kg^?1 (viscera), and 126.9?mg?kg^?1 (head without gills). After determining Cu²⁺, Zn²⁺, and Ca²⁺ by absorption spectrometry, a decrease in the Ca²⁺ concentration was noted in the head during the experimental period. These observations were associated with the occurrence of a decalcification in the bone tissue in the presence of Al³⁺. In contrast, it was found that Zn²⁺ ions may act as a protective agent against Al³⁺-induced contamination.     

Effects of cadmium on metallothionein and histology in gills of tilapia (Oreochromis niloticus) at different salinities

The objectives of this study were to evaluate the effects of sublethal cadmium concentrations on the levels of cadmium, metallothionein (MT) and histological changes in gills of East Java strain tilapia (Oreochromis niloticus) at different salinity levels. The levels of cadmium in control gills were not significantly different at 0, 5 and 10 practical salinity unit (PSU). The cadmium concentrations in gills of cadmium-exposed fish were significantly higher at 0 PSU than at 5, 10 and 15 PSU. The MT concentrations in control gills were not significantly different at 0, 5, and 10 PSU. The MT concentrations of cadmium-exposed fish were significantly higher than those in respective control groups at 0, 5 and 10 PSU. Significant gill damage occurred in fish exposed to cadmium at lower salinity. The epithelial lifting was noted at gills of fish exposed to 2.5 mg/L of cadmium at 0 PSU, and telangiectasis was observed at gills exposed to 5 mg/L of cadmium at 0 PSU. The level of gill damage decreased with increasing salinity of media. The increased MT and histological changes in gills of our findings could be a protective response of animals to toxic effect of cadmium.     

饲料中铜暴露对吉富罗非鱼幼鱼血红细胞微核和组织中铜蓄积的影响

为了研究饲料中铜暴露对吉富罗非鱼幼鱼的毒性作用,将1 080条罗非鱼幼鱼暴露于6个浓度梯度的高铜日粮中,通过60 d的暴露试验,测定罗非鱼血红细胞微核变化和组织中铜的蓄积.结果表明,铜在各组织中积累量的顺序依次为:肝脏＞脾脏＞肠道＞鳃＞肌肉,与对照组相比,其中肝脏、脾脏、肠道中铜的含量随着时间和日粮中铜浓度的升高而逐渐升高,而鳃和肌肉中铜含量无显著差异;罗非鱼幼鱼血红细胞微核率、核异常率、总核异常率与日粮中铜的浓度存在一定的剂量—效应关系,并且核异常率高于微核率.