Comet assay for the detection of genotoxicity in blood cells of flounder (Paralichthys olivaceus) exposed to sediments and polycyclic aromatic hydrocarbons     

Woo S  Kim S  Yum S  Yim UH  Lee TK 《Marine pollution bulletin》2006,52(12):1768-1775

To investigate the genotoxic effect of marine sediments on aquatic organism, sediment samples were collected from 13 sites along the coast of Gwangyang Bay (Korea). Concentrations of polycyclic aromatic hydrocarbons (PAHs) in sediments were determined and the relationship between exposure of flounder blood cells to sediment extracts and DNA single-strand breakage in the blood cells was examined using the comet assay. Levels of DNA damage were proportionally increased by exposure concentration and the highest sediment-associated DNA damage was observed at the station showing the highest PAHs contamination. DNA damage in blood cells exposed to five types of PAHs (benzo[a]pyrene, fluoranthene, anthracene, pyrene and phenanthrene) and in flounder (Paralichthys olivaceus) exposed to benzo[a]pyrene (BaP) for 0, 2 and 4 days were assessed by measuring comet tail length. The tail lengths of five PAHs-exposed groups at 50 and 100 ppb were significantly different from the non-exposed group, and the genotoxic effect of BaP correlated with both concentration and duration of exposure. Throughout the study, significant differences in DNA breakage were recorded between cells exposed to sediment extracts or PAHs and non-exposed control. This study demonstrated the comet assay as a successful tool in monitoring contamination of marine sediments and assessing genotoxicity of PAHs in marine organisms, either in vitro or in vivo.  相似文献   

牙鲆gnrh基因的表达分析     

彭壮壮  邹玉霞  刘琰  梁少帅  王雯祥  王丽娟  邹聪聪  周顺  尤锋 《海洋科学》2021,45(6):118-125

本文通过qPCR方法,研究了我国重要海水鱼类——牙鲆（Paralichthys olivaceus）雌、雄成体组织以及性腺分化期的脑、性腺中gnrh1、gnrh2和gnrh3的表达水平。结果显示,gnrh1分布于所有检测的组织：在脑、垂体、肾脏和肝脏中高表达,在性腺中表达相对较低;gnrh2则在雌、雄性的肝脏以及雄性的肠、肾脏、眼睛和头肾中;而gnrh3只在雄性的肾脏、眼睛和肌肉中检测到微弱表达。人工诱导的牙鲆雌核发育鱼苗分别用17β-雌二醇（E₂）和17α-甲基睾酮（MT）处理,使其分化为雌性或雄性个体表型。在性腺分化期的脑中,gnrh1的表达呈现上升趋势,在全长（total length,TL）4 cm鱼苗中,E₂组的表达显著高于MT组（P<0.05）;gnrh2在2 cm TL时E₂组显著高于MT组（P<0.05）,随后在E₂和对照组的表达水平均出现下降趋势;gnrh3在2 cm TL时E₂和对照组的表达均显著高于MT组（P<0.05）,自6 cm TL时MT和E₂组的表达开始下降。在性腺中,三种gnrh在性腺分化前2 cm TL表达量都相对较高,随后呈现下降趋势。综上,推测牙鲆gnrh1可能参与了性腺分化的启动、分化过程及性腺发育,gnrh2主要参与了性腺分化的启动,gnrh3主要参与了性腺分化的启动和分化过程。  相似文献   

牙鲆(Paralichthys olivaceus)与大菱鲆(Scophthalmus maximus)受精前期微管骨架的免疫荧光观察          下载免费PDF全文

朱香萍  尤锋  张培军  徐永立 《海洋与湖沼》2008,39(2):190-196

利用免疫荧光显微技术研究观察了牙鲆和大菱鲆正常二倍体受精卵从胚盘形成到4-细胞期微管骨架的动态变化过程,获得了一系列中心体、纺锤体及核的变化图像。牙鲆和大菱鲆正常发育的受精卵中微管骨架结构-纺锤体和中心体的动态变化过程基本相似,纺锤体及微管的形态结构也基本相似,大部分受精卵的微管骨架结构随卵裂呈现明显的周期性变化。在相同培育温度下,大菱鲆受精卵的胚盘形成时间要比牙鲆受精卵的胚盘形成时间晚10min左右,这与解剖镜下现场观察的结果相一致。首次利用免疫荧光显微技术从分子细胞生物学角度对牙鲆和大菱鲆进行细胞学观察,为进一步对其进行染色体操作提供分子细胞学依据。  相似文献   

牙鲆(Paralichthys olivaceus)外周血单个核细胞的离体培养和电生理特征研究          下载免费PDF全文

王彬  孙金生 《海洋与湖沼》2008,39(3):217-221

利用细胞分层液(比重为1.083)进行养殖牙鲆外周血单个核细胞(包括淋巴细胞和单核细胞)的分离、纯化及培养,并采用膜片钳技术观察了单核细胞膜的电生理特征.结果表明,牙鲆外周血单个核细胞在添加PHA(30(u)g/ml)的M199(20?S)培养液中,于25clC塑料培养(Costar,美国)中能够贴壁生长传至6代,存活30天以上.单核细胞呈不规则圆形,细胞长径为(12.30±0.62)μm,短径为(10.29±1.05)μm,并具有较强的吞噬活性,吞噬率(PP)和吞噬指数(PI)分别达到(61.9士3.05)%和1.58±0.51;淋巴细胞多呈圆形或椭圆形,细胞长径为(8.50±0.82)μm,短径为(7.46±1.07)μm,吞噬率和吞噬指数分别达到(42.1±2.33)%和3.17±1.57.膜片钳研究结果表明,牙鲆单核细胞适宜进行膜片钳研究,记录的细胞均表达电压门控钠、钾、钙离子通道.  相似文献   

Genetic variation of natural and cultured stocks of Paralichthys olivaceus by allozyme and RAPD     

尤锋  张培军  王可玲  相建海 《中国海洋湖沼学报》2007,25(1):78-84

Population genetics of the left-eyed flounder, Paralichthys olivaceus, including natural and cultured stocks distributed in the coastal waters near Qingdao of eastern maritime China, was analyzed in allozyme and RAPD. The results showed that among total 29 gene loci of 15 isozymes, 9 and 7 were po- lymorphic in natural and cultured stocks, respectively. The status of genetic diversity in P. olivaceus is low in terms of polymorphic loci in chi-square test and genetic departure index of Hardy-Weinberg equi- librium. More alleles in IDHP, CAT, GDH and Ldh-C allozymes were found in the fish, which could be used as markers in assortive breeding and distinguishing stock, population or species evolution. Total 88 and 86 RAPD bands ranging from 200 to 2 500 bp were recognized individually in average of 7.8–8.0 bands per primer. The genetic diversity in cultured stock is lower than that in natural ones showing an ob- viously decreasing genetic divergence. Therefore, effective countermeasures must be taken to protect ge- netic resources of marine cultured fishes. The 2 markers have their own pros and cons. Combining the 2 markers to investigate the genetic variation of populations is suggested. The results provide basic data of this flounder and they are useful for studying genetic improvement and genetic resources of the fish.  相似文献   

Carbon Budget of Bastard Halibut Paralichthys Oliva-eus in Relation to Body Weight and Temperature     

线薇薇  刘瑞玉  朱鑫华 《中国海洋湖沼学报》2003,21(2)

The effects of body weight and temperature on the carbon budget of the juvenile bastard halibut, Paralichthys olivaceus , were studied at temperature 13.5, 18, 21.5 and 24 ℃, respectively. The carbon intake, faecal and growth carbon were measured, and the carbon respiration was calculated using the carbon budget equation(CC=GC+FC+RC). The combined relationship between different components of the carbon budget, body weight and temperature could be described by regression equations: CC =1.0206 W 0.8126 e 0.1483 T; G=0.0042 W 1.4096 (-5.11 T 3+285.90 T2-5173.72 T +30314.03); FC =0.0485 W0.7711e 0.1624 T ; UC = 1.4333 W 0.6715e 0.1487 T. Body weight had no significant ffect on the carbon absorption efficiency and the conversion efficiency.  相似文献   

长山列岛邻近海域褐牙鲆资源丰度的时空分布及其与环境因子的关系     

徐晓卉  纪毓鹏  薛莹  张崇良  刘晓慧  任一平  徐宾铎 《海洋学报》2022,44(2):39-45

本文根据2016?2017年在长山列岛邻近海域开展的渔业资源和环境季度调查数据,利用两阶段广义可加模型分析了该海域褐牙鲆资源丰度的时空分布及其与相关因子之间的关系.结果表明,长山列岛邻近海域褐牙鲆资源丰度具有明显的季节变化,春、秋季资源丰度高于冬、夏季;褐牙鲆春、秋季主要分布在120.5°～122.5°E水域,夏、冬季...  相似文献   

静水压法人工诱导牙鲆(♀)×圆斑星鲽(♂)杂交三倍体的探索     

刘金相  王旭波  刘聪辉  高金宁  王忠凯  王文基  王志刚  于海洋  齐洁 《中国海洋大学学报(自然科学版)》2014,44(2)

开展了牙鲆(Paralichthys olivaceus)(♀)和圆斑星鲽(Verasper variegatus)(♂)杂交,通过静水压抑制受精卵第二极体的释放来诱导杂交三倍体,设计单因素分析实验,筛选出当前试验条件下的最佳诱导条件。结果表明:静水压法可成功诱导出杂交三倍体,诱导率主要受到受精后处理起始时刻、处理压强和压强处理时间3个因素的影响。静水压法的最佳诱导条件为:海水温度(15.0±0.2)℃,受精后3 min起,以70 MPa压力处理6 min。此时相对孵化率为(24.06±1.10)%,利用倍性分析仪对其进行倍性鉴定,三倍体率为(26.67±6.67)%。染色体分析表明,三倍体胚胎的染色体数目为71条,对照组二倍体胚胎的染色体数目为47条。还发现有少量染色体数目为24条的单倍体胚胎。研究结果为鲆鲽鱼类异源多倍体的培育提供了参考。  相似文献   

温度、光照及仔鱼个体大小对稚海蜇(Rhopilema esculentum)捕食褐牙鲆(Paralichthys olivaceus) 仔鱼的影响     

曹亮  于鑫  刘金虎  南鸥  窦硕增 《海洋与湖沼》2012,43(3):520-526

利用实验生态学方法,研究温度(15、19和23℃)、光照(明和暗)、仔鱼大小[全长(3.01±0.08)mm初孵仔鱼和(3.79±0.13)mm孵化后5天仔鱼]对稚海蜇[伞径(21.1±0.4)mm]捕食褐牙鲆仔鱼的影响。结果表明,稚海蜇对初孵仔鱼的捕食率随着水温升高而显著升高。光照条件在各仔鱼密度(10,30,50和80ind/L)下均不显著影响稚海蜇对初孵仔鱼的捕食率,说明光条件并不显著改善稚海蜇捕食仔鱼的能力。在高仔鱼密度(50和80ind/L)下,稚海蜇对孵化后5天仔鱼的捕食率显著低于对初孵仔鱼的捕食率;但在低密度(10和30ind/L)下,稚海蜇对两个不同发育阶段仔鱼的捕食率之间无显著差异。在低仔鱼密度下,稚海蜇对仔鱼的捕食率较低,这会降低仔鱼个体大小对稚海蜇捕食率作用的显著性。这既表明仔鱼逃避被稚海蜇捕食的能力随其发育生长而提高,也反映了稚海蜇对不同个体大小仔鱼捕食作用受仔鱼密度制约。  相似文献   

海豚链球菌3种免疫原对牙鲆血细胞吞噬及SOD、POD和ACP活力的影响     

赵娜  唐小千  绳秀珍  战文斌 《中国海洋大学学报(自然科学版)》2012,42(4):33-40

本文制备海豚链球菌(Streptococcus iniae)强毒株NUF849的福尔马林灭活全菌(FKC)、胞外产物(ECPs)以及全菌与胞外产物的混合物(FKC+ ECPs)3种免疫原,以其免疫牙鲆(Paralichthys olivaceus),并在免疫牙鲆后第42天分别以海豚链球菌NUF849和NUF812攻毒.在免疫牙鲆后第7、14、21、28、35、42天及攻毒后第7天分别取样,测定血细胞吞噬能力及血清中超氧化物歧化酶(SOD)、过氧化物酶(POD)和酸性磷酸酶(ACP)的活力,以比较3种免疫原对这些免疫指标的影响.结果显示,免疫后尤其攻毒后,红细胞、淋巴细胞、单核细胞、嗜中性粒细胞等的吞噬率与吞噬指数均上升,FKC+ ECPs组显著高于其他组(P＜0.05);各免疫组SOD活力随时间延长而下降并趋于稳定,攻毒后各组数值有不同程度的升高,但FKC+ ECPs组数值最低;免疫组POD活力先升高,第28天起下降,攻毒后各组均下降,FKC+ ECPs组始终高于其他免疫组(P＜0.05);免疫组ACP活力也在免疫后第28天达到峰值,以某株NUF849攻毒后FKC+ ECPs组水平上升(相对于免疫后第42天),其他组显著下降;FKC、ECPs、FKC+ ECPs对菌株NUF849的相对免疫保护宰分别为50％、40％和90％,对NUF812的相对免疫保护率分别为60％、30％和80％,FKC+ ECPs组显著高于其他组(P＜0.05).以上研究表明,各免疫组均诱导了牙鲆的非特异性免疫应答,具有一定的免疫保护效果,其中FKC+ECPs组效果最佳.  相似文献