Relationships between quality parameters and content of glycerol galactoside and porphyra-334 in dried laver nori <Emphasis Type="Italic">Porphyra yezoensis</Emphasis>

ABSTRACT:   Glycerol galactoside (GG; floridoside + isofloridoside) and porphyra-334 (P-334) are contained in nori (Susabinori Porphyra yezoensis and Asakusanori Porphyra tenera ). Glycerol galactoside has been found to have bifidogenic growth stimulator activity and P-334 is known to have ultraviolet-absorbing activity in the UVA region of sunlight. These substances have, respectively, potential for application to pre-biotic foods and in cosmetics as a sunscreen. In the present study, to investigate the relationships between GG and P-334 contents and the quality of nori, we measured the GG and P-334 contents with other components (total protein, chlorophyll-a, β-carotene and phycobillins) that are related to the quality of nori samples produced from different production areas and with different qualities. We found that the GG content was closely negatively correlated with the contents of other components, whereas P-334 was positively correlated with the other components. From these results, it is suggested that low-quality nori is a potential source of GG, and as a source for P-334, scraps of nori produced during nori processing should be suitable.     

大黄鱼mtDNA ND5和Cytb基因的克隆与序列分析

2004年4月，将采自浙江省象山港海区网箱养殖的大黄鱼样本，提取总DNA，通过设计特异性引物对大黄鱼线粒体DNA（mtDNA）的辅酶5（ND5）和细胞色素b（Cytb）基因进行PCR扩增。扩增产物经琼脂糖电泳检测、纯化后直接测序。得到ND5的序列1839 bp和Cytb基因序列382 bp。应用primer premier5和MEGA3软件包所作的系统发育分析表明：依据大黄鱼ND5序列所作的进化树总体支持传统的分类地位，大黄鱼更接近塘鳢鱼科。而基于Cytb基因所作的分析表明，黑鳃梅童鱼是大黄鱼在石首鱼科中是遗传距离最小的。     

LHRH-A和5-HT拮抗剂的协同作用对草鱼鱼种生长激素分泌活动和生长的影响

促性腺激素释放激素(LHRH)能够促进鱼类生长激素(GH)的释放,而5-羟色胺(5-HT)抑制鱼类GH的释放。将促性腺激素释放激素类似物(D-Ala6-Pro9-NEt-LHRH,LHRH-A,5μg/g饵料)和5-HT的特异性拮抗剂盐酸米安色林(Mianserin hydrochloride MIS,0.5 mg/g饵料)拌在饵料中投喂草鱼(Ctenopharyngodon idellus)鱼种6周,可促使血清GH水平显著提高,并使食物转化率升高,生长速率加快,肥满度增加;且两者促生长的协同作用比单独使用LHRH-A或MIS好。对草鱼鱼种体质和主要生化组成的分析表明,LHRH-A和MIS的促生长作用既没有引起鱼体可食部分的蛋白质含量减少,也不会使脂肪、水分含量和内脏质量增加,因而其促进的是草鱼种苗均衡、正常的生长。由此说明在生产应用中,使用高活性的神经内分泌因子,通过拌饵方式投喂对于促进养殖鱼类苗种的生长、缩短养殖周期,是有效而且安全的。     

镜鲤脂肪酸延长酶5基因的克隆和表达分析

脂肪酸延长酶5(ELOVL5)是高不饱和脂肪酸(HUFA)合成的关键酶之一。为了研究鲤HUFA合成的能力和机制,本研究采用RT-PCR和RACE技术获得镜鲤ELOVL5全长c DNA序列。ELOVL5基因c DNA全长1 121 bp,开放阅读框为876 bp,编码291个氨基酸。序列分析显示,镜鲤ELOVL5氨基酸序列包含1个组氨酸簇(HXXHH),1个典型的内质网驻留信号,多个跨膜区域和多个保守区域(KXXEXXDT、QXXFLHXYHH、NXXXHXXMYXYY、TXXQXXQ),具有典型的脂肪酸延长酶的结构特征。氨基酸同源性分析结果显示,镜鲤ELOVL5基因与其他鱼类同源性为79.0%~93.1%,与人同源性为69.0%。通过实时荧光定量PCR(RT-q PCR)检测该基因在镜鲤不同组织中的表达量,发现脂肪酸延长酶基因在镜鲤肝中表达量最高,其次为脑,在背部肌肉中表达量最低。镜鲤ELOVL5基因的获得为进一步研究镜鲤HUFA的合成途径及调控机理奠定了基础。     

草鱼C5a、C5aR和FⅡ多克隆抗体的制备与表达特性

为研究草鱼补体蛋白5a (complement component 5a,C5a)、补体蛋白5a受体(C5a receptor,C5aR)和凝血因子Ⅱ(blood coagulation factorⅡ,FⅡ)在感染草鱼呼肠孤病毒(grass carp reovirus,GCRV)时的蛋白表达和相互作用,针对草鱼C5a和FⅡ蛋白构建了原核表达体系、针对草鱼C5aR蛋白构建C5aR-KLH偶联物,纯化蛋白,免疫日本大耳白兔制备3种蛋白的多克隆抗体。用蛋白质印迹(Western blot)、免疫共沉淀(Co-IP)和拉下(pull down)实验检测3种蛋白表达与互作关系,Western blot结果显示,C5a和C5aR在健康草鱼的肝脏、脾脏、肾脏、头肾、肠、鳃和肌肉中均有蛋白表达,FⅡ在肝脏、脾脏和肠中表达,而在肾脏、头肾、鳃和肌肉中不表达;在感染GCRV的草鱼肝脏组织中C5a、C5aR和FⅡ蛋白均随病程进展呈现上升趋势。Co-IP结果显示,在GCRV处理后,C5a、C5aR和FⅡ蛋白具有相互作用关系。pull down结果显示,C5a pull down共鉴定得到C3、RIG-I等2...     

凡纳滨对虾养殖塘主要有机污染指标相关性分析

2009年4—9月期间,对上海市奉贤区某凡纳滨对虾(Litopenaeus vannamei)养殖场池塘两个养殖周期中总有机碳(TOC)、高锰酸盐指数(CODMn)、和五日生化需氧量(BOD5)进行分析检测。结果显示:试验的两个周期中(4—7月、7—9月),14个池塘BOD5分别为(8.62±3.08)mg/L和(10.47±3.87)mg/L,CODMn为(13.09±3.98)mg/L和(16.16±6.07)mg/L,TOC为(17.60±5.91)mg/L和(20.32±6.07)mg/L。TOC/CODMn分别为(1.35±0.22)和(1.32±0.30);TOC/BOD5为2.10±0.44和2.08±0.63;BOD5/CODMn为0.66±0.13和0.65±0.11。TOC、CODMn和BOD5两两之间呈显著线性正相关,第一个养殖周期中:BOD5=0.4174TOC+1.2777,r=0.8022;CODMn=0.5616TOC+3.2091,r=0.8342;BOD5=0.6264CODMn+0.4209,r=0.8106。第二个养殖周期中:BOD5=0.4764TOC+0.7902,r=0.7480;CODMn=0.7941TOC+0.0237,r=0.7962;BOD5=0.568CODMn+1.2912,r=0.8920。结果表明,利用建立的TOC、CODMn和BOD5之间的相关方程,可进行各指标之间的相互换算,从而与不同的水质标准或其他相关研究进行对比,为凡纳滨对虾养殖池塘的水质管理和健康养殖提供指导。     

鹅源H5N1亚型禽流感病毒新疆分离株A/Goose/XJYL/10/2003HA基因的克隆和序列测定与分析

根据已发表的H5N1亚型禽流感病毒的HA基因全序列,设计了1对引物,对禽流感病毒新疆株A/Goose/XJYL/10/2003(H5N1)提取RNA,经RT-PCR扩增后将其克隆到pMD18-T载体上,获得长为1758bp的HA全基因序列。序列测定后,分析结果表明,本试验株与广东、香港和东南亚等不同地区的H5N1禽流感病毒HA的核甘酸序列的同源率很高,其氨基酸切割位点附近具有高致病性禽流感病毒的特征。系统进化树分析表明,新疆株为独立分支,属于欧亚种系。     

水声学探测在江河鱼类资源评估中的技术分析

水声学探测已成为鱼类资源评估的重要手段之一。介绍了水声学调查的方案设计和鱼类资源量估算方法,并结合2009年冬季三峡库区水声学调查,着重阐述EY60回声探测系统的正确使用方法和Sonar5分析软件的应用,探讨水声学技术用于江河鱼类资源调查存在的不足以及应用展望,旨在为水声学设备及分析软件的正确使用提供技术参考。     

Measurement of 5-day biochemical oxygen demand without sample dilution or bacterial and nutrient enhancement

A direct method for measuring the 5-day biochemical oxygen demand (BOD₅) of aquaculture samples that does not require sample dilution or bacterial and nutrient enrichment was evaluated. The regression coefficient (R²) between the direct method and the standard method for the analyses of 32 samples from catfish ponds was 0.996. The slope of the regression line did not differ from 1.0 or the Y-intercept from 0.0 at P = 0.05. Thus, there was almost perfect agreement between the two methods. The control limits (three standard deviations of the mean) for a standard solution containing 15 mg/L each of glutamic acid and glucose were 17.4 and 20.4 mg/L. The precision of the two methods, based on eight replicate analyses of four pond water samples did not differ at P = 0.05.     

Characterization of mevalonate metabolism in the sea bass (Dicentrarchus labrax L) liver

The activities of mevalonate kinase, mevalonate 5-phosphate kinase and mevalonate 5-pyrophosphate decarboxylase, were examined in sea bass (Dicentrarchus labrax L) liver. The activities of the three enzymes were studiedin vitro in relation to the influence of protein content, time of incubation, pH, temperature, mevalonate, ATP and Mg⁺⁺ concentration. Protein content in the assay medium affected the three enzymes differently. Mevalonate kinase, mevalonate 5-phosphate kinase, and mevalonate 5-pyrophosphate decarboxylase activities were linear up to 0.2, 0.4 and 0.8 mg protein, respectively. With respect to the time course studies, the enzymes also behaved differently. Mevalonate kinase activity increased over forty minutes, reaching a plateau thereafter, while mevalonate 5-phosphate kinase and decarboxylase increased over the entire assay period. All the three enzymes showed a maximum in activity at pH 7.5. The effect of reaction temperature showed that phosphorylation increased to maximum around 35°C for mevalonate kinase and 30°C for mevalonate 5-phosphate kinase while decarboxylation rates remained constant well until 30°C temperature decreasing afterwards. The enzymes behaved differently as a function of mevalonate concentration. Mevalonate 5-phosphate formed was maximal when the initial mevalonate concentration was 272 M, whereas mevalonate 5-pyrophosphate and CO₂ were formed maximally at mevalonate concentrations of 136 M and 68M, respectively. Optimal ATP concentration in the medium was 3 mM for decarboxylase and 6 mM for kinases, and Mg⁺⁺ requirements varied from 4 mM for decarboxylase to 6 mM for kinases.     

克氏原螯虾不同发育时期5-HT与DA在卵巢和肝胰腺中的分布

为了研究5-HT与DA阳性物质在克氏原螯虾(Procambrus clarkii)不同发育时期的卵巢细胞和肝胰腺细胞中的分布特征,运用免疫组化法对卵巢发育4个不同时期的卵巢与肝胰腺进行观察。结果表明,各期卵巢卵细胞和肝胰腺细胞中均存在5-HT与DA阳性物质。Ⅰ期卵巢5-HT阳性物质分布于卵母细胞胞核,胞质未出现阳性;Ⅱ期～Ⅳ期5-HT阳性物质主要集中在卵母细胞胞质以及胞核。在发育各期的肝胰腺中5-HT的免疫物质均呈阳性。Ⅰ～Ⅳ期卵巢中,DA阳性物质主要分布于卵母细胞的胞核以及肝胰腺细胞,其强度呈递减趋势。结果提示5-HT具有促进卵巢发育作用,而DA则抑制卵巢发育。     

不同规格瘤背石磺胃盲囊5-羟色胺和P物质分布与数量的初步研究

将不同规格野生瘤背石磺,按8g以下、8-18g和18g以上分为三组,组织切片观察胃盲囊组织结构,并采用免疫组化方法,观察5-羟色胺和P物质在胃盲囊中分布及数量变化规律。结果表明：瘤背石磺胃盲囊主要由非纺锤型和纺锤型两种类型绒毛组成,这两种绒毛所占比例在瘤背石磺不同发育阶段明显不同,在18g以上组中以纺锤型为主,此时纺锤型所占比例(66%)约为非纺锤型(34%)的2倍。各组瘤背石磺胃盲囊中均有大量5-HT和P物质阳性细胞的分布,这些细胞主要分布于上皮粘膜层,且主要分布在非纺锤型绒毛中,5-羟色胺和P物质阳性非纺锤型绒毛比例均匀约为纺锤型绒毛的10倍。5-羟色胺和P物质阳性细胞数在瘤背石磺胃盲囊的分布规律,均有随瘤背石磺不断发育重量不断增加而增加的趋势,其中18g以上组5-羟色胺阳性细胞数（112.82个·cm-2）约为8g以下组（34.8个·cm-2）的3倍,而18g以上组P物质阳性细胞数（177.48个·cm-2）约为8g以下组（51.08个·cm-2）的3.5倍。在各组中P物质阳性细胞的数量明显高于5-羟色胺,且在不同发育段其增加趋势更为明显。结论：瘤背石磺胃盲囊5-羟色胺和P物质分布均有随体重增加而增加的趋势。这些细胞主要分布于非纺锤绒毛的粘膜上皮。其中P物质阳性细胞数明显高于5-羟色胺。     

Effect of feeding and fasting on plasma tryptophan and tryptophan to large neutral amino acid ratio,and on brain serotonin turnover in rainbow trout,Oncorhynchus mykiss

Two time-course experiments were conducted to determine the effect of feeding and fasting on the plasma ratio of tryptophan (trp) to the large neutral amino acids (LNAA), (trp/LNAA ratio) and brain serotonin (5-HT) turnover in rainbow trout,Oncorhynchus mykiss. Trout were fasted overnight or for 3 days and were then either fed or continued to be fasted for up to a further 3 days. Changes in plasma trp, plasma trp/LNAA ratio, brain trp, brain 5-HT, brain 5-hydroxyindoleacetic acid (5-HIAA) and brain 5-HIAA/5-HT ratio were determined over time. Feeding decreased the plasma trp/LNAA ratio, brain trp and the brain 5-HIAA/5-HT ratio. In addition, in fish sampled over 3 days, there appeared to be a rhythm in plasma trp and the brain 5-HIAA/5-HT ratio which was independent of feeding. These results indicate that in rainbow trout, feeding is a sufficient physiological event to decrease brain 5-HT turnover. Furthermore, feeding-independent changes in the brain 5-HIAA/5-HT ratio, which were evident in fasted fish sampled over 3 days, also suggest an additional, non-feeding-related modulator(s) of brain 5-HT turnover in rainbow trout.     

水库网箱养鱼与水环境保护关系的研究

通过连续2年进行不投铒网箱养鱼试验，并每2个月对网箱养殖区及上、下游进行水体理化监测，结果表明，网箱养鱼能改善水体透明度，降低BOD5、CODMn含量，并对降低TP也有一定作用。     

基于ND4和ND5基因序列研究八种臂尾轮虫的系统关系和分类地位

通过对尾突臂尾轮虫、裂足臂尾轮虫、剪形臂尾轮虫、方形臂尾轮虫、红臂尾轮虫、镰形臂尾轮虫、矩形臂尾轮虫和十指臂尾轮虫8种臂尾轮虫的ND4和ND5基因序列进行扩增、测序,结合Gen Bank数据库中的褶皱臂尾轮虫、萼花臂尾轮虫、转轮虫和旋轮虫的ND4和ND5基因序列,使用MEGA6.0软件构建这12种轮虫系统发生树(NJ树、ME树、UPGMA树),探讨了8种臂尾轮虫之间的系统关系。结果显示,本研究所涉及的轮虫ND4和ND5基因序列差异百分比均值为40.3%,可作为分子标记应用于轮虫属内种间系统关系研究;系统树所显示亲缘关系与形态学研究基本一致,均支持将十指臂尾轮虫、裂足臂尾轮虫归入臂尾轮属。     

三疣梭子蟹Drosha和Exportin 5基因克隆及其在性腺发育过程中的表达分析

本研究利用RACE技术克隆了三疣梭子蟹(Portunus trituberculatus)Drosha和Exportin 5基因的cDNA全长序列,Drosha基因长度为3443 bp,编码1038个氨基酸,预测蛋白质包含2个相邻的RNA酶Ⅲ结构域(RⅢDa和RⅢDb)和1个双链RNA结合结构域(dsRBD)。Exportin 5基因全长为5000 bp,编码1208个氨基酸,预测蛋白质包含1个Importin-βN-末端结构域(IBN-N)和1个核输出蛋白结构域(XPO-1)。同源分析显示,三疣梭子蟹Drosha氨基酸序列与其他物种高度相似,与日本对虾(Marsupenaeus japonicus)相似度最高,达94%。qRT-PCR分析结果显示,Drosha和Exportin 5基因在三疣梭子蟹各组织中均有表达,其在卵巢和肝胰腺中的表达量最高(P0.05);在精巢不同发育时期,2个基因表达量的变化趋势一致,均在Ⅱ期(精母细胞增殖、分化期)表达量最高;在卵巢发育过程中,2个基因表达量的变化趋势也大致相同,随着卵巢发育(Ⅱ~Ⅴ期)逐渐升高。研究表明,Drosha和Exportin 5基因可能通过调控miRNA的合成来影响三疣梭子蟹性腺发育过程,为深入解析三疣梭子蟹性腺发育调控机制提供了重要参考。     

Genetic identification and phylogenetic inferences in different Mugilidae species using 5S rDNA markers

A DNA methodology was developed to discriminate fry of six Mugilidae species found in the Mediterranean, namely Mugil cephalus, Mugil so‐iuy, Chelon labrosus, Liza aurata, Liza ramada and Liza saliens. Polymerase chain reaction (PCR) amplification of the 5S rDNA gene was used for the identification of the above six species. PCR products of two species showed different patterns on EtBr‐stained agarose gels; M. so‐iuy gave a pattern of three bands, while L. saliens gave a pattern of one band. Mugil cephalus, C. labrosus, L. aurata and L. ramada gave a pattern of two bands. Subsequent sequencing analyses revealed unique haplotypes for each of the remaining four species. This is a genetic technique that could be applied in hatcheries, for identification of fry mullet's species. Furthermore, 5S rDNA sequences of each of five of the studied species (i.e. M. cephalus, C. labrosus, L. aurata, L. ramada and L. saliens), as well as a sequence of Oedalechilus labeo that belongs to the Mugilidae family, were aligned and used for the examination of the phylogenetic relationships among them. Phylogenetic trees produced in our study are in general agreement with those presented in the literature. This is the first study to use a nuclear marker to elucidate the phylogeny of Mugilidae species.     

核糖体蛋白S5(RPS5)基因研究进展

核糖体蛋白S5(ribosomal protein S5,RPS5)是真核生物核糖体的重要组成部分,目前对RPS5功能的研究主要侧重于以下几方面:RPS5的晶体结构及其在核糖体结构上的分布特点;RPS5与蛋白质的翻译;RPS5具有DNA连接蛋白Ⅱ的功能;RPS5具有转录延伸因子3(EF-3)的功能;RPS5与物种进化;RPS5突变体的研究。本文对RPS5基因的研究概况及功能进行综述。     

中国对虾ATG5基因的克隆及其在pH、碳酸盐碱度胁迫下的表达分析

采用cDNA末端快速扩增(RACE)技术获得中国对虾(Fenneropenaeus chinensis) ATG5基因cDNA全长,命名为FcATG5。利用实时荧光定量技术分析了FcATG5的组织表达及其在pH和碳酸盐碱度胁迫下的表达特征,并利用RNAi技术验证其功能。基因分析显示,FcATG5 cDNA全长为2225 bp,开放阅读框为810 bp,编码269个氨基酸,预测其编码的蛋白质分子量为31.103 kDa,理论等电点为5.59,为疏水性蛋白,包含1个APG5自噬相关蛋白结构域,无跨膜结构,不包含信号肽。同源性和系统进化分析显示,FcATG5具有高度保守性,与凡纳滨对虾(Litopenaeus vannamei)同源性最高(98.14%)。组织表达分析显示,FcATG5在中国对虾各组织中均有表达,肌肉中表达量最高(P<0.05),血淋巴细胞中最低(P<0.05)。pH胁迫后48 h,FcATG5在鳃中的表达量最低,为对照组的1.68倍;胁迫后96 h最高,为对照组的2.67倍。碳酸盐碱度胁迫后12 h,FcATG5在鳃中表达量最高,为对照组的2.77倍;胁迫后96 h最低,为对照组的1.30倍。干扰实验结果显示,pH和碳酸盐碱度胁迫下,沉默FcATG5基因会使中国对虾死亡率显著增高(P<0.05),表明该基因的表达量越高,越有利于中国对虾存活。实时定量结果表明,FcATG5在pH、碳酸盐胁迫下的表达量均显著升高(P<0.05),推测自噬可能参与中国对虾应对非生物胁迫的调控。本研究结果对水生动物特别是甲壳动物中细胞自噬研究具有重要的借鉴意义,有助于推进中国对虾盐碱水养殖的研究进程。