双酚A 对斑马鱼胚胎发育阶段的毒性及生物蓄积

为研究胚胎在不同发育阶段对外源化合物的毒性敏感差异,以0hpf、8hpf 和8h 终止3 种暴露方式进行染毒,探讨斑马鱼胚胎在不同暴露时段对双酚A(BPA)毒性和生物蓄积特征.结果表明,0hpf 暴露时,斑马鱼胚胎内BPA 含量与暴露浓度(0,5.00,10.00,15.00mg/L)及暴露时间(0,8,24,32,48,72h)呈明显正相关关系;在相同BPA 浓度(15.00mg/L)作用下,斑马鱼胚胎在0hpf 暴露下BPA 富集量比8hpf 的大,表现为致死与亚致死毒性都大,证明斑马鱼胚胎在发育早期(受精后到原肠胚期)对BPA 污染更为敏感;8h 终止试验进一步证明原肠胚期前的斑马鱼胚胎对BPA 的这种敏感特征.     

nano-ZnO对斑马鱼鱼鳔发育毒性效应研究

根据鱼鳔发育的不同阶段,从受精后0 h(0 hours post fertilization, 0 hpf)开始对5个发育阶段(0 hpf～孵化前、0～120 hpf、0～168 hpf、孵化后～120 hpf、120～168 hpf)的斑马鱼进行nano-ZnO暴露实验,研究不同浓度的nano-ZnO对斑马鱼鱼鳔的影响.结果表明,nano-ZnO悬浮液中溶解的Zn~(2+)不是导致斑马鱼死亡和鱼鳔缺损的唯一或主要原因.nano-ZnO暴露浓度越高,斑马鱼的死亡率和鱼鳔缺损率越高,存在剂量-效应关系.15 mg·L~(-1) nano-ZnO可造成90%的斑马鱼鱼鳔缺损和死亡,10 mg·L~(-1) nano-ZnO可导致斑马鱼鱼鳔面积缩小70%.斑马鱼鱼鳔早期发育阶段(0～168 hpf)对nano-ZnO敏感度大小为:出芽阶段(0 hpf～孵化前)充气阶段(孵化后～120 hpf)充气完成阶段(120～168 hpf).斑马鱼鱼鳔的发育面积和发育时间也受nano-ZnO暴露染毒的影响而发生变化.斑马鱼死亡率和鱼鳔缺损率(鱼鳔发育)之间存在相关性(r=0.978,p0.01).这表明斑马鱼鱼鳔的发育缺损是造成斑马鱼胚胎死亡的主要原因之一,但nano-ZnO对斑马鱼鱼鳔的影响机理,尤其是对鱼鳔出芽阶段和充气阶段的致毒机理需要进一步研究.     

三氯卡班环境暴露对斑马鱼神经行为的影响

为了探究三氯卡班(TCC)对脊椎生物神经行为的干扰及机制, 以脊椎模式生物斑马鱼为研究对象, 分别评判了TCC亚致死剂量急性暴露对幼鱼的神经发育毒性及慢性暴露对成鱼神经行为的影响.结果表明: TCC亚致死剂量急性暴露造成斑马鱼胚胎及幼鱼自主运动活力减少, 对声光刺激的敏感程度下降.吖啶橙凋亡染色表明, TCC诱导幼鱼头肾、脑部和眼睛等多部位发生细胞过度凋亡.通过扫描电镜观察, 侧线神经丘形态畸形, 发育受到抑制, 毛细胞团数量减少且排列紊乱.成鱼行为学试验结果显示, TCC长期暴露的成鱼过度焦虑及恐慌, 避险和警惕能力下降, 并表现出自闭, 不愿社交, 显著影响斑马鱼的记忆力、学习能力和辨别认知功能.     

高氯酸钠对斑马鱼胚胎的毒性效应

采用斑马鱼胚胎发育技术方法研究高氯酸钠对斑马鱼胚胎的急性毒性效应,配制一系列高氯酸钠的浓度,对受精1h的胚胎进行染毒暴露,最后对斑马鱼胚胎的心率,孵化率,死亡率,畸形率进行统计分析。研究结果显示,高氯酸钠(NaClO4)浓度的增加,斑马鱼胚胎的心率和孵化率降低、死亡率和畸形率增加。斑马鱼48hpf的胚胎心率高浓度组比对照组下降了35%;72 h的孵化率由对照组的88%降低到高浓度组的36%;144 h的死亡率由对照组的5%增加到72%;144 h的畸形率由对照组的2%增加到91%,主要畸形表现为心包囊肿(PE),脊柱弯曲(AM),鱼鳔缺失(SBD)等。结果表明,高浓度的高氯酸钠(NaClO4)对斑马鱼胚胎发育有明显的剂量效应作用。     

CdSe/ZnS量子点对斑马鱼胚胎发育的毒性效应

本研究以模式生物斑马鱼(Danio rerio)为对象,观察在不同浓度Cd Se/Zn S量子点(Cd Se/Zn S QDs)暴露下,斑马鱼胚胎形态发育、氧化应激以及金属硫蛋白MT基因和应激蛋白Hsp70基因表达变化情况.结果表明,Cd Se/Zn S QDs对斑马鱼胚胎72 hpf(hours post fertilization)的半致畸效应浓度(EC50)为316.994 nmol·L-1.QDs暴露影响了斑马鱼胚胎的死亡率、畸形率、孵化率、自主运动频率和体长,以及引起胚胎卵凝集,心包囊肿,脊椎弯曲等多种毒性效应;同时导致斑马鱼体内超氧化物歧化酶(SOD)活性变化以及丙二醛(MDA)含量增加.QDs还诱导斑马鱼MT基因和Hsp70基因表达上调,斑马鱼机体产生一系列自我保护反应来减轻QDs所造成的伤害.这表明:Cd Se/Zn S QDs对斑马鱼胚胎产生了毒性效应,其毒性可能与其核心Cd2+的释放、粒径大小以及氧化应激有关.     

五氯酚对稀有鮈鲫胚胎毒性效应研究

研究五氯酚(PCP)对稀有鮈鲫(Gobiocypris rarus)胚胎的致畸和毒性效应.以7.5,30,60,120,250μg/L5个浓度的PCP对0hpf(hpf,受精卵孵出时间)的稀有鮈鲫胚胎进行暴露染毒,同时设置空白对照组、二甲基亚砜溶剂对照组和雌二醇(EE2,2.5ng/L)阳性对照组.在立体显微镜下观察整个胚胎的发育过程,统计胚胎的孵化率、96hpf相对存活率和各时期的畸形率,并利用半定量RT-PCR检测胚胎中CYP1A基因和p53基因mRNA的表达.结果表明,PCP暴露能延迟稀有鮈鲫胚胎发育,并造成胚胎卵凝结、心包囊肿、脊柱弯曲等多种畸形甚至死亡.随着PCP暴露浓度的升高,稀有鮈鲫胚胎的孵化率和96hpf相对存活率降低,各时期的畸形率增加,并呈现一定的浓度效应.稀有鮈鲫胚胎CYP1A基因和p53基因mRNA表达被显著诱导,并随PCP浓度的升高而增加.PCP对稀有鮈鲫胚胎发育表现为显著的毒性效应.稀有鮈鲫胚胎孵化率、96hpf相对存活率、各时期畸形率及CYP1A基因和p53基因的诱导表达可以作为评价PCP毒性作用的敏感指标.     

伏杀硫磷对斑马鱼心脏发育毒性及氧化损伤机制

以斑马鱼为研究对象,评价伏杀硫磷急性暴露对幼鱼心脏发育毒性及氧化损伤作用.结果表明,1和3mg/L的伏杀硫磷导致斑马鱼胚胎孵化率分别下降10.37%和22.96%,体长分别缩短11.77%和19.40%,心率显著下降,产生心包水肿等胚胎发育畸形.抗氧化酶(SOD,CAT)活性检测显示伏杀硫磷造成斑马鱼幼鱼体内SOD活性随浓度增加先上升后下降,CAT活性显著下降;活性氧(ROS)检测表明伏杀硫磷对幼鱼的心脏部位造成氧化损伤;Tg(cmlc2:EGFP)转基因斑马鱼证明伏杀硫磷诱导幼鱼的心房和心室过度分离;吖啶橙(AO)染色显示幼鱼心脏部位发生细胞过度凋亡;心脏发育相关的基因中nppa和sox9b显著上调,gata4和vmhc显著下调;凋亡相关基因bcl2、bax、Puma、Mdm2的表达均显著下调.伏杀硫磷暴露造成的心脏发育毒性与氧化损伤和细胞凋亡平衡的破坏相关.     

双酚AF暴露对胚胎期和幼鱼期斑马鱼的毒性效应

为明确不同发育阶段的斑马鱼对BPAF(双酚AF)暴露的易感性,采用暴露试验法初步研究了BPAF对胚胎期和幼鱼期斑马鱼的发育毒性. 结果表明:①BPAF暴露可延缓胚胎期斑马鱼的发育和孵化,使其出现心包水肿、卵黄囊异常、心率下降、心脏搏动停止等症状. ②暴露96 h后,ρ(BPAF)为2.0、2.5、3.0 mg/L暴露组胚胎期斑马鱼的畸形率高达100%,暴露24 h致畸的EC₅₀(半数效应浓度)为2.00 mg/L,暴露96 h的LC₅₀(半数致死浓度)为1.84 mg/L. ③暴露72 h后,对照组幼鱼期斑马鱼鱼鳔发育缺陷率为0;除ρ(BPAF)为1.0 mg/L暴露组外,其余BPAF暴露组幼鱼期斑马鱼鱼鳔发育缺陷率为100%. ④随着暴露时间的延长以及中毒程度的加深,幼鱼期斑马鱼出现的中毒症状依次表现为心包水肿、卵黄囊水肿、背脊弯曲、心跳停止,暴露48和72 h时其心包水肿的EC₅₀分别为1.76、1.56 mg/L,暴露96 h的LC₅₀为1.77 mg/L. 胚胎期和幼鱼期斑马鱼对BPAF暴露响应的差异分析显示,幼鱼期斑马鱼对BPAF暴露的反应更为敏感,幼鱼期斑马鱼的心包水肿症状可作为BPAF毒性响应的最佳指标之一.      

氯苯和间甲酚对斑马鱼胚胎和仔鱼联合毒理效应研究

方法:采用斑马鱼胚胎和仔鱼发育技术,分别用不同浓度的氯苯和间甲酚混合液对斑马鱼胚胎和仔鱼进行暴露试验。结果:两种受试物联合对斑马鱼胚胎和仔鱼发育表现不同的联合毒性。0hpf染毒主要表现为:对24 h胚胎尾部延展为拮抗作用,48 h水肿、72 h孵化和畸形均为协同作用;48 h对仔鱼毒性为由相加作用向拮抗作用的转变,在24 h、72 h和96 h联合毒性均为协同作用。结论:氯苯和间甲酚联合作用对斑马鱼胚胎和仔鱼存在毒性。     

2,4-二氯酚和2,4,6-三氯酚对斑马鱼胚胎发育的影响

为评价2,4-二氯酚(2,4-DCP)和2,4,6三氯酚(2,4,6-TCP)对鱼类早期生命阶段的影响,以斑马鱼胚胎为研究对象,观察2,4-DCP和2,4,6-TCP对96 h内斑马鱼胚胎发育的影响。结果表明,两种氯酚对斑马鱼胚胎发育均有抑制作用,可以造成胚胎孵化延迟,降低体长及心率,幼鱼畸形甚至死亡;2,4-DCP、2,4,6-TCP的NOEC分别为1.12 mg/L和0.50 mg/L。两种氯酚对斑马鱼胚胎的发育毒性相比,不同观察指标,毒性不尽相同:斑马鱼胚胎48 h心率指标对2,4-DCP更加敏感,而斑马鱼胚胎96 h孵化率、畸形率、体长对2,4,6-TCP更加敏感。     

氯苯对斑马鱼胚胎发育和仔鱼的毒性效应研究

选取健康的雌雄斑马鱼按1:1或1:2的比例进行交配产卵。采用静态法,以丙酮为助溶剂,分别用不同浓度的CB进行斑马鱼受精卵和刚平衡游动仔鱼的暴露试验,记录胚胎孵化数和胚胎、仔鱼畸形数以及仔鱼死亡数。不同浓度的CB可导致斑马鱼胚胎孵化率降低及孵出的仔鱼畸形率增加,且毒性呈现剂量-效应关系。当CB浓度高于0.88μg/L时,斑马鱼胚胎及其孵出的仔鱼畸形率开始增加;当CB为108.9μg/L时,暴露24 h的胚胎出现全致畸效应。能平衡游动的斑马仔鱼对CB的毒性也很敏感,24 h、72 h和96 h的LC50值分别为95.35、80.51和62.28μg/L;110.0μg/L CB使处理24 h的仔鱼全部死亡。较低浓度的CB就使斑马鱼胚胎发育畸形;其对仔鱼的毒性也很显著,达到一定浓度可使其死亡。     

氟吡菌胺对斑马鱼的毒性效应

采用斑马鱼胚胎发育技术和传统毒理学方法, 研究了氟吡菌胺对斑马鱼胚胎、成鱼及仔鱼的毒性效应.结果发现,氟吡菌胺对3个阶段斑马鱼均具有致死能力,对仔鱼LC50(48h)值为0.204mg/L,成鱼LC50(96h)为0.286mg/L,幼鱼LC50(96h)为1.489mg/L.研究表明,0.0596mg/L以上浓度的氟吡菌胺对斑马鱼胚胎均有一定程度的致死效应,高浓度处理组胚胎出现心包囊肿、卵黄囊不吸收、黑色素沉积少及鱼体弯曲等症状;氟吡菌胺对斑马鱼成鱼进行14d暴露后,处理组成鱼体重下降,产卵量减少,有效卵量降低; 0.0298mg/L处理组144h子代仔鱼存活率虽然比较高,但存活鱼大部分出现心包囊肿、体弯曲等畸形.上述结果说明,水体中残留的氟吡菌胺对于斑马鱼各生命阶段的生长发育均具有潜在的危害.     

Pheromonal regulation of starvation resistance in honey bee workers (<Emphasis Type="Italic">Apis mellifera</Emphasis>)

Most animals can modulate nutrient storage pathways according to changing environmental conditions, but in honey bees nutrient storage is also modulated according to changing behavioral tasks within a colony. Specifically, bees involved in brood care (nurses) have higher lipid stores in their abdominal fat bodies than forager bees. Pheromone communication plays an important role in regulating honey bee behavior and physiology. In particular, queen mandibular pheromone (QMP) slows the transition from nursing to foraging. We tested the effects of QMP exposure on starvation resistance, lipid storage, and gene expression in the fat bodies of worker bees. We found that indeed QMP-treated bees survived much longer compared to control bees when starved and also had higher lipid levels. Expression of vitellogenin RNA, which encodes a yolk protein that is found at higher levels in nurses than foragers, was also higher in the fat bodies of QMP-treated bees. No differences were observed in expression of genes involved in insulin signaling pathways, which are associated with nutrient storage and metabolism in a variety of species; thus, other mechanisms may be involved in increasing the lipid stores. These studies demonstrate that pheromone exposure can modify nutrient storage pathways and fat body gene expression in honey bees and suggest that chemical communication and social interactions play an important role in altering metabolic pathways.     

Toxic effects of perfluorononanoic acid on the development of Zebrafish (Danio rerio) embryos

Perfluorononanoic acid (PFNA) is a nine-carbon perfluoroalkyl acid widely used in industrial and domestic products. It is a persistent organic pollutant found in the environment as well as in the tissues of humans and wildlife. There is a concern that this chemical might be a developmental toxicant and teratogen in various ecosystems. In the present study, the toxic effects of PFNA were evaluated in zebrafish (Danio rerio) embryos. One hour post-fertilization embryos were treated with 0, 25, 50, 100, 200, 300, 350, and 400 μmol/L PFNA for 96 hr in 6-well plates. Developmental phenotypes and hatching rates were observed and recorded. Nineteen genes related to oxidative stress and lipid metabolism were examined using Quantitative RT-PCR and confirmed by whole mount in situ hybridization (WISH). Results showed that PFNA delayed the development of zebrafish embryos, reduced the hatching rate, and caused ventricular edema and malformation of the spine. In addition, the amount of reactive oxygen species in the embryo bodies increased significantly after exposure to PFNA compared with that of the control group. The Quantitative RT-PCR and WISH experiments demonstrated that mRNA expression of the lfabp and ucp2 genes increased significantly while that of sod1 and mt-nd1 decreased significantly after PFNA exposure. The mRNA expression levels of gpx1 and mt-atp6 decreased significantly in the high concentration group. However, the mRNA expression levels of both ppara and pparg did not show any significant variation after exposure. These findings suggest that PFNA affected the development of zebrafish embryos at relatively low concentrations.