两种铜(Ⅱ)-司帕沙星配合物与DNA作用及生物毒性比较

选择铜(Ⅱ)-司帕沙星-邻菲啰啉Cu(sf)phenCl(sf代表司帕沙星负离子,phen代表邻菲啰啉)和铜(Ⅱ)-司帕沙星Cu(sf)22种配合物,通过电子吸收光谱法、荧光分析法及琼脂糖凝胶电泳法研究了其与DNA的作用,首次以斑马鱼胚胎为模式生物,研究了它们的生物毒性,并对2种配合物的研究结果进行了比较。光谱实验结果表明:Cu(sf)phenCl与DNA的作用强于Cu(sf)2;琼脂糖凝胶电泳实验结果显示:在抗坏血酸存在条件下,Cu(sf)phenCl对pBR322质粒DNA的切割作用更强;同时,斑马鱼胚胎毒性实验显示Cu(sf)phenCl对斑马鱼胚胎毒性也更大。     

斑马鱼(Danio rerio)胚胎免疫反应: 补体基因在早期胚胎以及LPS 刺激胚胎中的表达

以模式生物斑马鱼(Danio rerio)为研究对象, 使用原位杂交及实时荧光定量PCR 方法, 对 斑马鱼补体基因(C3, C4, C9, Bf1, Bf2, Bf3)及其调节因子(RCA2.1, RCA2.2)在早期胚胎中的时空表达 模式及LPS 刺激后表达量的变化进行了研究。结果表明, 上述基因在胚胎早期(卵裂期至体节期)均 为泛表达, 从受精后24h 至幼鱼阶段特异性表达于肝脏及消化道等器官。对胚胎显微注射LPS 后, C3、C9、Bf1、Bf2、Bf3 基因在早期胚胎中表达上调, RCA2.1 基因表达下调, 这与斑马鱼成鱼受到 革兰氏阴性菌感染后补体分子(C3, C9, B 因子等)的反应类似, 表明其参与补体激活途径、溶膜途径 及补体调节, 提示斑马鱼可在胚胎发育早期构建补体系统, 参与急性期反应等免疫反应。     

贻贝发育早期酯酶和过氧化物酶活性

采用组织化学和分光光度技术对贻贝(Mytilus edulis)发育早期酯酶(Esterase,EST)和过氧化物酶(Per-oxidase,POD)活性进行了研究,以探讨贝类早期发育过程中的免疫防御机制。EST和POD均存在于贻贝的卵母细胞、胚胎和幼虫中,但在各个发育时期的分布和含量不同。组织化学定位结果表明,从卵母细胞到D型面盘幼虫各个发育时期的胚胎和幼虫均呈EST强阳性和POD阳性;卵母细胞核区较大且颜色较浅;桑椹期开始出现不同细胞阳性的强弱差异;原肠胚、担轮幼虫和D型面盘幼虫细胞间阳性强弱的差异逐步增大。分光光度测定结果表明,α-醋酸酯酶活力,卵母细胞最低,受精卵略有提高,卵裂期胚胎提高较大且达最高峰,囊胚、原肠胚和担轮幼虫又逐渐下降,D型面盘幼虫下降较大;α-丁酸酯酶活力,卵母细胞较高,受精卵略有提高且达到最高峰,卵裂期胚胎到D型面盘幼虫各个时期逐渐下降,D型面盘幼虫最低。不依赖卤素POD活力,卵母细胞最低,受精卵提高较大,囊胚达到最高峰,原肠胚、担轮幼虫和D型面盘幼虫逐渐降低。依赖卤素POD活力与不依赖卤素POD活力的变化规律相似,但受精卵提高较小,而卵裂期提高较大。EST和POD可能在贻贝的发育早期抵抗病原微生物侵染方面发挥重要作用。     

Effects of salinity on embryonic and larval development of Chinese mitten crab Eriocheir sinensis(Decapoda: Brachyura) and salinity-induced physiological changes

To investigate the effects of salinity on early development of Chinese mitten crab ( Eriocheir sinensis ), and the salinity tolerance mechanism of embryos, different developmental stages of embryos (gastrula, eyespot and pre-hatching stage), and hatched stage I zoea and megalopa, were exposed to a range of salinities (1, 5, 10, 15 (control), 20, 25, 30, 35 and 40). Hatching, survival and molting were monitored. Effects of 24-hour hypersaline (35) and hyposaline (1) stress on egg diameter, water content, Na +/K +-ATPase (NKA) activity, and crustacean hyperglycemic hormone (CHH) gene mRNA expression in embryos and megalopa, are reported. Embryos are more tolerant of low (≤ 5) than high (≥25) salinities, with optimum ranges for gastrula and pre-hatching stage embryos being 5-20, and for eyespot embryo and stage I zoea, 10-20. Most megalopa can molt to the first juvenile instar by day 5 at salinities between 1 and 40, whereas molting of megalopa stages was delayed at 40. Hypersaline conditions resulted in a loss of moisture, reduction of egg volume, and a signifi cant increase in NKA activity and CHH mRNA expression at some developmental stages. Hyposaline conditions did not affect moisture content or egg volume, but resulted in decreased NKA activity and CHH mRNA expression in embryos. For megalopa stages, NKA activity was significantly upregulated following both hypo- and hypersaline stress. Our results suggest high salinity will inhibit development and hatching of E. sinensis embryos, and low salinity will affect the survival of their stage I zoea. Increased NKA activity in embryos and megalopa stages might indicate a hyporegulation response under hypersaline conditions. These findings provide useful information for spawning ground protection of indigenous E . sinensis and enrich the knowledge of embryonic tolerance mechanisms of hyperregulating crustaceans following osmotic stress.     

鮸状黄姑鱼的早期发育研究

本文对状黄姑鱼的胚胎和仔、稚鱼各发育阶段的形态特征进行了描述。绝状黄姑鱼的受精卵为圆球形浮性卵,无色透明。在水温20～23℃,盐度26～30的条件下,经24h35min陆续孵出仔鱼。胚胎发育分为26期。初孵仔鱼至孵化后4日龄为仔鱼前期;孵化后5～22日龄为仔鱼后期;孵化后23～32日龄为雅鱼期;孵化后33日龄进入幼鱼早期。仔、稚鱼培育水温18～28C,盐度20～30。仔鱼2日龄稍开口,3～4日龄为混合营养阶段,是仔鱼的主要危险期。     

温度、盐度对半滑舌鳎胚胎发育的影响

温度及盐度是鱼类苗种生产当中2个重要的因子。研究了在不同水温、盐度条件下,半滑舌鳎胚胎发育的情况。通过对孵化时间、孵化率、初孵仔鱼畸形率、活力的比较,发现温度对受精卵的孵化影响显著。受精卵在18℃以下及28℃以上均不能孵化出仔鱼,在18~28℃水温条件下,受精卵可以孵化,其适宜的孵化水温为22~23℃,且随着温度升高受精卵孵化时间逐渐缩短。在水温23℃下,受精卵在盐度16以下及48以上均不能孵化出仔鱼,在盐度18~46的范围内,受精卵均可以孵化出仔鱼,盐度38以上均为畸形,且孵化时间不受盐度的影响,其适宜孵化盐度为28~32。     

泥螺的繁殖生物学研究

研究了泥螺的繁殖方式，繁殖季节，繁殖力，受精卵的孵化，幼虫发育及变态和稚，幼螺的发育过程，泥螺在浙江沿海每年的繁殖期为3-11月，5-6月和9-10月为繁殖高峰。繁殖期内亲螺有一次交配多次产卵现象，平均每个亲螺产卵3-4次，每次产卵群1个，每个卵群平均含有受精卵6282个，个体繁殖力18848.4-25128.0粒，在水温28.5℃条件下，面盘幼虫孵化时间为72h，孵化率为93.8%，幼虫经过2-8d浮游生活后进入匍匐生活，28-30d后发育变态成为稚螺，50d后成为幼螺。     

重金属对波部东风螺早期发育的毒性及EDTA的解毒效果

研究了重金属Cu2 、Zn2 、Cd2 、Pb2 对波部东风螺(Babyloniaformosaehabei)胚胎和面盘幼体的毒性效应及EDTA对幼体Cu2 、Zn2 中毒的解毒效果。结果表明,Cu2 、Zn2 、Cd2 、Pb2 在胚胎发育中最高允许浓度分别为0.01、0.05、0.10、0.50mg·L-1;对幼体的48hLC50值分别为33.251、86.352、1524.191、4751.042μg·L-1,96hLC50值分别为21.772、59.418、1072.182、3031.019μg·L-1;安全浓度分别为2.177、5.942、107.218、303.102μg·L-1;4种重金属离子对胚胎和幼体的毒性顺序均为:Cu2 >Zn2 >Cd2 >Pb2 。Cu2 、Zn2 浓度分别为0.2、0.5mg·L-1时,EDTA对面盘幼体的最佳解毒浓度分别为1—2和2—3mg·L-1。     

苜蓿组织培养体细胞胚发生的细胞学观察

苜蓿无菌苗下胚轴在含２ｍｇ／Ｌ２．４－Ｄ和０．５ｍｇ／Ｌ６ＢＡ的ＭＳ培养基中诱导出愈伤组织，愈伤组织在含有３ｍｇ／ＬＮＡＡ和１．５ｍｇ／ＬＫＴ的ＳＨ培养基中形成胚性愈伤组织，并伴随有体细胞胚的发生。显微观察表明：体细胞胚起源于单个的胚性细胞，它既能在愈伤组织内部发生，又可在其表面发生。体细胞胚经过球形、心形、鱼雷形及子叶胚各阶段可形成再生植株，这为在细胞水平上对苜蓿进行遗传操作及苜蓿品种的改良成为可能。     

A field method for the concurrent measurement of fine sediment content and embryo survival in artificial salmonid redds

The infiltration cube method presented in this paper allows concurrent field measurement of fine sediment content and embryo survival in an incubating environment reproducing, as closely as possible, the morphology and grain size composition of a natural salmonid redd. An infiltration cube is made up of a rectangular (30 cm square and 20 cm high) steel frame with no walls, which has a folded plastic bag attached to its base. Each infiltration cube is buried in the substrate using a procedure aimed at reproducing, as closely as possible, the construction and morpho‐sedimentological structure of a natural salmonid redd. Using a tripod installed on the river bed or at the surface of the ice, the infiltration cube method provides a quick, easy and robust way to pull relatively large samples (approximately 65 kg) out of underwater substrate even during tough winter field conditions. The absence of walls on the cube also precludes bias in infiltration rates by both allowing lateral sediment transport within substrate and preventing loss of fine particles during retrieval from underwater. Copyright © 2006 John Wiley & Sons, Ltd.