玉米赤霉烯酮暴露导致的秀丽隐杆线虫生殖及发育毒性的世代间比较

利用模式生物秀丽隐杆线虫(Caenorhabditis elegans,C.elegans)对玉米赤霉烯酮(zearalenone,ZEN)暴露导致的生殖、发育缺陷在世代间的可传递性进行了研究。对亲代染毒,后代断毒恢复,实验结果显示,ZEN暴露导致的体长发育缺陷可在线虫后代中得到恢复,而生殖毒性可产生严重缺陷,且这种缺陷在后代中没有明显的恢复。通过多代持续染毒实验,发现ZEN对亲代P0、子一代F1和子二代F2线虫产生的生殖和发育毒性存在差异,其毒性作用强度为P0F2F1。线虫世代之间的发育缺陷和生殖缺陷表现出逐代修复的规律,但修复程度不同,推测与线虫自身修复的复杂应激活动有关。     

不同尺度纳米镍对秀丽线虫的发育毒性

应用模式生物-秀丽隐杆线虫研究金属纳米镍的生殖发育毒性.采用20 nm和90 nm镍分别以2.5和5.0 μg·cm2两个剂量对秀丽隐杆线虫进行染毒,并以微米镍和生理盐水作对照,采用后代数目、世代时间、形态和体长、寿命和半数致死率等生殖发育相关评价指标,对纳米镍生殖发育毒性进行评价.结果发现,与生理盐水对照组和微米镍组比较,秀丽线虫暴露于20 nm和90 nm镍的两个剂量组后,均表现出生殖和发育的异常(P)＜0.01),并有剂量依赖性.且90 nm镍暴露对秀丽线虫后代数目、世代时间、形态和体长、寿命和半数致死天数等指标的缺陷作用均显著于20 nm镍.研究表明,纳米镍可影响秀丽线虫的生殖和发育功能,这一效应与纳米粒径和暴露浓度有关.此结论可为制定纳米镍的接触限值标准提供参考.     

不同尺度纳米镍对秀丽线虫发育毒性的比较研究

应用模式生物-秀丽隐杆线虫研究金属纳米镍的生殖发育毒性。采用20nm和90nm镍分别以2.5和5.0μg·cm-2两个剂量对秀丽隐杆线虫进行染毒,并以微米镍和生理盐水作对照,采用后代数目、世代时间、形态和体长、寿命和半数致死率等生殖发育相关评价指标,对纳米镍生殖发育毒性进行评价。结果发现,与生理盐水对照组和微米镍组比较,秀丽线虫暴露于20nm和90nm镍的两个剂量组后,均表现出生殖和发育的异常(P<0.01),并有剂量依赖性。且90nm镍暴露对秀丽线虫后代数目、世代时间、形态和体长、寿命和半数致死天数等指标的缺陷作用均显著于20nm镍。研究表明,纳米镍可影响秀丽线虫的生殖和发育功能,这一效应与纳米粒径和暴露浓度有关。此结论可为制定纳米镍的接触限值标准提供参考。     

脱氧雪腐镰刀菌烯醇、黄曲霉毒素B_1和玉米赤霉烯酮对秀丽隐杆线虫的联合毒性研究

脱氧雪腐镰刀菌烯醇(DON)、黄曲霉毒素B_1(AFB_1)和玉米赤霉烯酮(ZEN)是最为常见的粮食真菌毒素,易共存于谷物产品和动物饲料中,而目前对其联合毒性的研究较少,且研究结果并不完全一致。为探究DON、AFB_1和ZEN的联合毒性作用,本论文以秀丽隐杆线虫(C.elegans)为模型,分别评估了毒素混合物AFB_1+DON、AFB_1+ZEN、DON+ZEN和AFB_1+DON+ZEN对C.elegans的生长发育(体长)和生殖能力(产卵量)的毒性作用,并用Chou-Talalay模型来判定毒素混合物的相互作用类型。研究表明,AFB_1、DON和ZEN单独染毒C.elegans时,其毒作用强弱为AFB_1ZENDON。联合染毒时,AFB_1+DON对C.elegans产生协同作用,而DON+ZEN则产生拮抗作用;AFB_1+ZEN对体长(24 h)和产卵量的毒作用随着暴露浓度的增加,由弱拮抗变为协同作用,而在毒素暴露48 h后,对线虫的生长发育呈协同作用;AFB_1+DON+ZEN除在EC50-24 h和EC75-24 h时对体长产生明显的毒性增强作用外,其他普遍表现出拮抗作用。由此表明,DON、AFB_1和ZEN对C.elegans的联合毒性作用与剂量和时间相关。     

砷与农药草甘膦、敌敌畏对秀丽隐杆线虫的联合毒性

农田常遭受砷和农药污染,这2种污染会产生联合毒性,影响生物体健康。利用野生型秀丽隐杆线虫进行砷、草甘膦和敌敌畏的24h单一毒性和联合毒性实验。结果表明,砷、草甘膦和敌敌畏对秀丽隐杆线虫24h-LC50分别为130.5mg·L-1、18.5mg·L-1、3.3μL·L-1;通过等效毒性相加法实验,发现砷与草甘膦对秀丽隐杆线虫的联合毒性为协同作用,砷与敌敌畏对秀丽隐杆线虫的联合毒性为拮抗作用。     

基于彗星实验的洛克沙胂对秀丽隐杆线虫胚胎细胞DNA损伤的研究

为评价洛克沙胂的遗传毒性,采用单细胞凝胶电泳(彗星实验),研究了不同浓度的洛克沙胂对秀丽隐杆线虫胚胎细胞脱氧核醣核酸(DNA)的损伤作用。提取秀丽隐杆线虫的胚胎细胞,分别暴露于0(空白对照)、50、250、500μg·L~(-1)含洛克沙胂的溶液染毒1 h。用彗尾DNA百分比含量(TDNA%)、彗星尾长(TL)和Olive尾矩(OTM)作为DNA损伤的指标。实验结果表明,与空白对照组比较,处理组中彗尾DNA百分比含量、彗星尾长以及Olive尾矩显著增加(P0.01)。随着洛克沙胂浓度的增加,彗尾DNA百分比含量、彗星尾长以及Olive尾矩逐渐增加,其相关系数r0.99,说明在实验浓度范围内,存在极显著的浓度-效应关系。洛克沙胂对秀丽隐杆线虫胚胎细胞DNA具有损伤作用,彗星实验操作简便、快速、灵敏度高,能够反映出洛克沙胂的遗传毒性。因此,通过彗星实验建立实验室检测洛克沙胂遗传毒性的方法具有可行性。     

重组人金属硫蛋白-Ⅲα短肽毒性效应的初步研究

研究重组人金属硫蛋白Ⅲα短肽(rh-MT-Ⅲα)致HaCaT细胞、L929细胞和秀丽隐杆线虫(Caenorhabditis elegans,C.ele-gans)毒性效应,为rh-MT-Ⅲα的安全应用提供实验基础.细胞暴露于不同浓度的rh-MT-Ⅲα溶液(0～400μg·mL-1),24h后检测细胞存活率和乳酸脱氢酶(LDH)活性,观察细胞形态并检测细胞凋亡情况.结果表明,各剂量组2种细胞的存活率,与对照组相比均无显著性差异,细胞形态也未观察到显著变化.在400μg·mL-1浓度下,LDH释放率明显增加,细胞凋亡率上升.线虫实验中,将线虫培养并同步化至L1期后进行染毒,剂量为5、50和500 μg·mL-1,暴露48 h后检测其运动行为(头部摆动和身体弯曲频率)、发育(体长)及摄食行为(咽泵频率),各指标均未有明显改变.研究表明,在一定的剂量水平下,rh-MT-Ⅲα对HaCaT细胞、L929细胞和秀丽隐杆线虫无明显毒性效应.     

2种新烟碱类杀虫剂对秀丽隐杆线虫神经毒性研究

新烟碱类(neonicotinoids)是一种类似于烟碱的内吸性杀虫剂,选择性作用于昆虫的中枢神经系统。目前,有关新烟碱类杀虫剂对非靶标生物的生态风险引发了全球的广泛关注。本文采用土壤生物秀丽隐杆线虫(Caenorhabditis elegans)作为受试生物,探究2种新烟碱类杀虫剂吡虫啉(imidacloprid)和呋虫胺(dinotefuran) 24 h暴露(1μg·L~(-1)～10 mg·L~(-1))后,对秀丽线虫运动行为、摄食行为以及乙酰胆碱酶(ACh E)活性和相关基因(ace,mtl)转录水平的影响。结果表明,2种新烟碱类杀虫剂均对秀丽线虫的生理指标具有随着浓度增加而抑制的效应,其中头部摆动是最为敏感的生理指标,10μg·L~(-1)吡虫啉暴露水平和100μg·L~(-1)呋虫胺暴露水平时,线虫头摆频率相较于对照组有显著减少(P 0.01),其次是身体弯曲摄食AChE活性(吡虫啉)和摄食AChE活性身体弯曲(呋虫胺),相关基因转录水平没有发现与浓度具有相关的效应,但在环境浓度下的暴露有明显的变化。新烟碱类杀虫剂对秀丽线虫的神经毒性作用表明了其对土壤中非靶标生物具有一定的生态风险。     

空气负离子暴露对秀丽隐杆线虫（Caenorhabditis elegans）的生物学效应

利用模式生物秀丽隐杆线虫(Caenorhabditis elegans)研究了电晕法空气负离子(Electrically generated negative air ions,ENIs)暴露的生物学效应.将成年野生型和mev-1突变型线虫及新鲜虫卵持续暴露于空气负离子中(空气负离子剂量1×105～1.5×105ions·cm-3),观察空气负离子暴露对线虫发育周期、寿命、繁殖性能和性腺细胞凋亡的影响.结果表明:空气负离子暴露可显著加速野生型和mev-1突变型线虫发育(p<0.05),缩短寿命(p<0.05),与野生型相比,空气负离子暴露对mev-1突变线虫寿命影响更大(p<0.05);空气负离子暴露可显著降低线虫后代数量(p<0.05),原因可能在于空气负离子暴露显著诱导性腺细胞凋亡的增加(p<0.05).以上结果表明,持续空气负离子暴露对线虫具有损伤作用,mev-1基因突变导致线虫对空气负离子暴露更加敏感.     

农药对秀丽隐杆线虫毒性效应及其机制的研究进展

随着大量农药被广泛的使用并最终汇聚到环境介质中,对生态环境和人体健康产生潜在影响。秀丽隐杆线虫(Caenorhabditis elegans)是土壤中最丰富的后生动物,在土壤生态系统中具有重要地位,并作为一种重要的模式生物广泛应用于环境毒理学研究。本文从秀丽隐杆线虫常用的毒理学研究方法入手,以致死率、生长发育、运动行为、繁殖、活性氧自由基(ROS)水平、细胞凋亡水平、相关基因表达量和蛋白水平等作为测试指标,归纳总结农药对秀丽隐杆线虫的衰老性、神经及生殖系统的毒性效应,分析探索其毒性机理,并展望了未来的研究重点。     

环境浓度下壬基酚对秀丽隐杆线虫时间依赖性的毒性兴奋效应

为研究暴露时间对壬基酚生态毒理学效应的影响,选取秀丽隐杆线虫(Caenorhabditis elegans)作为受试生物,采用急性(24 h和72 h)、慢性(10 d) 3个不同时间点,进行壬基酚环境浓度下(0、0.0001、0.001、0.01、0.1、0.2 mg·L~(-1))的暴露实验,以秀丽隐杆线虫的生理指标(体长、运动行为)、生化指标(氧化应激、细胞凋亡、脂褐素)为终点进行评估。结果表明:急性暴露24 h后,活性氧自由基(ROS)在0.01 mg·L~(-1)时即出现了明显的下降,其他生理生化指标无统计学意义上的明显变化。72 h暴露条件下,体长随壬基酚浓度的增加呈现倒U型趋势,在0.0001 mg·L-1时表现出最大刺激效应,比对照组高出26.4%(P0.01);头部摆动频率和身体弯曲频率在2个较高浓度(0.1和0.2 mg·L~(-1))时表现出明显的刺激效应;细胞凋亡水平在0.0001 mg·L~(-1)时显著性下降,呈现负剂量-效应关系(P0.05),在0.2 mg·L~(-1)时表现出最大刺激效应,比对照组降低了45.5%; ROS及脂褐素在壬基酚暴露浓度范围内呈正相关性增加。慢性暴露条件下,脂褐素在0.001 mg·L~(-1)时表现出最大效应,和对照组相比下降了65.4%,并且随着壬基酚浓度的增加呈现U型趋势;其他生理生化指标随着壬基酚浓度的增加表现出明显的负面效应。研究揭示了环境浓度水平的壬基酚对秀丽隐杆线虫的生态毒理效应是时间依赖性的,急性暴露以引起刺激作用为主,表现为毒性兴奋效应,而长期暴露后壬基酚对生理生化指标的负面效应更为明显。上述结果为进行壬基酚的毒性评价及更好地理解其毒性作用机理提供基础数据。     

GO通过氧化损伤激活秀丽线虫未折叠蛋白反应

以秀丽线虫为受试生物,探讨氧化石墨烯(GO)对秀丽线虫未折叠蛋白应答的激活及其与氧化应激的关系。将L4幼虫暴露于GO中24 h,通过检测活性氧(ROS)水平、乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等指标评估GO对秀丽线虫所致氧化损伤;以hsp-4和hsp-6分别作为秀丽线虫内质网和线粒体未折叠蛋白反应(UPR)的报告基因评价GO对秀丽线虫UPR应答的激活;同时以谷胱甘肽(GSH)作为抗氧化剂探讨GO所致UPR应答与氧化应激的关系。研究结果显示,GO暴露后秀丽线虫体内ROS、LDH和MDA水平升高,抗氧化酶活性上升;内质网和线粒体UPR反应增强,GSH预处理可以降低内质网和线粒体UPR应答。研究表明,GO短期暴露可以诱导秀丽线虫机体氧化应激进而激活UPR应答,抗氧化保护可以降低UPR应答反应。     

纳米二氧化硅对秀丽线虫的毒性作用研究

为探讨二氧化硅纳米颗粒(SiNP)的毒性作用.以微米SiO2和空白做对照,将尺寸为20和60 nm的SiNP分别以0.2,1.0和5.0 g·L-13个浓度对模式生物秀丽隐杆线虫进行染毒.通过第一代和第二代线虫体长、身体弯曲频率、头部摆动频率以及第一代线虫后代数目、世代时间等发育、运动和生殖的相关评价指标,对SiNP毒...     

Effects of temperature and food ration on gonad growth and oogenesis of the green sea urchin, Strongylocentrotus droebachiensis

To assess the effects of both temperature and food ration on gonad growth and oogenesis of the green sea urchin, Strongylocentrotus droebachiensis (O.F. Müller), individuals collected December 1996 (winter experiment) and June 1997 (summer experiment) were maintained for 3 months in one of four experimental treatments: (1) 3 °C and fed ad libitum (high ration), (2) 3 °C and fed one-seventh of the maximum ration (low ration), and (3) 12 °C and fed the high ration; (4) 12 °C and fed the low ration. All individuals were fed an artificial diet and exposed to only 1 h of light every day. At the end of both experiments, mean gonad indices of sea urchins fed the high ration had increased significantly (11–24% and 6–19% in the winter and summer experiments, respectively), while the gonad indices of individuals fed the low ration did not change. At the high ration (both experiments), the increase in gonad index of sea urchins occurred primarily as the result of a significant increase in the mass of nutritive phagocytes, as revealed by histological analyses. Primary oocytes were significantly larger in individuals held at 3 °C than at 12 °C throughout the winter experiment, regardless of food ration; during the summer experiment, primary oocytes were significantly larger in individuals receiving the high ration, regardless of the temperature at which they were held. These results suggest that: (1) food availability is the most important factor regulating energy storage and the relative size of gonads throughout the year, (2) temperature affects the rate of growth and maturation of primary oocytes during the later stages of oogenesis, and (3) once gametogenesis has been initiated, mature ova will be produced, even under conditions of low food availability. Conditions of high food availability in summer and low temperature in winter would thus favor reproductive output in sea urchin populations. Received: 1 March 2000 / Accepted: 4 October 2000     

Sexual reproduction of the solitary sunset cup coral Leptopsammia pruvoti (Scleractinia, Dendrophylliidae) in the Mediterranean. 2. Quantitative aspects of the annual reproductive cycle

There are few data on the reproductive biology of coral species living in temperate zones, particularly in the Mediterranean Sea. Leptopsammia pruvoti is a solitary coral that is commonly found in sea caves and under overhangs throughout the Mediterranean basin and along European coasts from Portugal to southern England. In this paper, we describe its annual reproductive cycle in the eastern Ligurian Sea near the city of Leghorn (Tuscany, Italy). Polyps were sexually mature at 3 mm in length (maximum diameter of the oral disc), were dioecious with a sex ratio of 1:1, and brooded their larvae. The maturation of spermaries took 12 months and oocytes 24 months. The rate of gonad development increased significantly from November to January, fertilization occurred from January to April and planulation during May and June. Seasonal variations in water temperature and photoperiod may have played an important role in regulating reproductive events. The amount of energy devoted to male gametogenesis (quantified by gonad index) was significantly higher in the sex separated species L. pruvoti than in the hermaphroditic dendrophylliid Balanophyllia europaea, whose reproduction has been studied in a previous work. We hypothesize that this difference is due to the contrasting sexuality or fertilization biology of these two species (cross-fertilization in the sex separated L. pruvoti versus possibly self-fertilization in the hermaphroditic B. europaea). Greater male sexual allocation in reproductive strategies characterized by dioecism or cross-fertilization when compared to those characterized by hermaphroditism or self-fertilization is common in plant mating systems. In relation to other solitary dendrophylliids, L. pruvoti presents an ‘r’-reproductive strategy.     

Annual reproductive cycle of the chiton Acanthopleura japonica

Annual reproductive cycle of the chiton Acanthopleura japonica (Lischke) was studied by observing both gonad index and histological features from October 1980 to October 1981. No significant changes in each factor were observed from October to April. Increased values of gonad index and spermatogenesis or vitellogenesis in gondas were observed from May to September. In the same time, gonad index decreased and sperms or oocytes in gonalds disappeared in the days around spring tide, and they were recovered around neap tide. Suggested spawning around spring tide from May to September is also supported by the occurrence of released eggs in the days around the new and full moons.     

Reproductive development of the barnacle Chthamalus malayensis in Hong Kong: implications for the life-history patterns of barnacles on seasonal, tropical shores

Hong Kong, lying just below the Tropic of Cancer (22°17′N, 114°09′E), experiences a strongly seasonal environment, with a cool almost temperate winter and a hot, tropical, summer. Histological sectioning of the gonads of the high-shore barnacle, Chthamalus malayensis Pilsbry, showed a seasonal trend in the development of its reproductive organs. Four stages of female gonad development were identified according to the cell types present: post-spawning, resting, growth and mature stages. The female gonad was mature from April to November, which was related to seawater temperatures, and entered a resting phase from December to March. Although the male gonad showed a seasonal developmental trend and reached maximum maturity in summer, the seminal vesicles were full of spermatozoa and functional throughout the year. The reproductive season of this species is therefore solely dependant upon the maturity of the female gonad. The estimated maximum number of broods per year was up to 10 and the maximum number of eggs produced per brood can reach 3,000 eggs. The minimum size for female gonad maturity was 6 mm rostro-carinal diameter (RCD) at which size, the barnacles were ~6-month old. Sperm production occurred at a smaller size (2 mm=2-month old). Compared with Chthamalus montagui and Chthamalus stellatus from temperate regions, C. malayensis produced a greater number of broods per year, had a longer reproductive period and faster gonad development. Chthamaloid barnacles in tropical regions may, therefore, invest more energy per year in reproduction during their life span. Contrary to the seasonal gonad developmental pattern of C. malayensis in the present study, however, C. malayensis in Singapore (which experiences only slight seasonal variation) had mature female and male gonads throughout the year, further supporting the strong role of climatic conditions effecting the reproductive biology of barnacles.     

Observations on the reproductive cycles of Monodonta lineata,Gibbula umbilicalis and G. cineraria

Monthly samples of gonads of 3 common species of intertidal trochids, Monodonta lineata (da Costa), Gibbula umbilicalis (da Costa) and G. cineraria (L.) were fixed for examination by light microscopy. Numbers and proportions of immature and mature oocytes in sections of ovaries were determined. In M. lineata and G. umbilicalis, the annual reproductive cycles were very similar, the testis and ovary of both species being empty brown structures from September to January/February, when gametogenesis began and the gonads began to swell and turn green in both sexes. As ripening continued, the ovary remained green in G. umbilicalis, but in M. lineata turned greygreen by the end of May. The testis of both species changed, to yellowish in G. umbilicalis and creamy-pink in M. lineata. Spawning occurred throughout summer and early autumn, and the gonads reverted to their former empty state. These changes were matched by the increase and subsequent decrease in proportion of ripe oocytes in the gonad. Some ripe oocytes are not shed during the spawning season, but no evidence of cytolysis of such oocytes was found. In G. cineraria, in contrast, the ovary remained green and swollen and the testis remained creamy-white and packed with sperm throughout the year. The proportion of ripe to unripe oocytes in the ovary fluctuated for the entire sampling period, and did not indicate the time of spawning. In the testis of all three species, spermatogonia were rarely seen, and spermatid stages were uncommon.