High prevalence of “Candidatus Rickettsia amblyommii” in Amblyomma ticks from a Spotted Fever Endemic Region in North Argentina

Ticks from an endemic Spotted Fever region in Argentina were analysed by PCR for Spotted Fever Group Rickettsiae. DNA of “Candidatus Rickettsia amblyommii” was found in 21.3% of Amblyomma hadanii and in 44.0% of A. neumanni. Amblyomma sculptum (formerly A. cajennense) and Haemaphysalis juxtakochi were negative for rickettsial DNA. DNA of Rickettsia rickettsii, the etiological agent of the clinical cases reported within the studied region was not detected in the analysed sample.     

营养因素对比目鱼形变的影响

营养因素是影响比目鱼形变的主要因素。本文介绍了比目鱼的形变并分析了轮虫和桡足动物的主要营养成分的差异,并着重介绍了甲状腺激素、维生素A、脂肪酸和能量对比目鱼着色和眼睛移位的影响。     

Effects of plastic mulches on root zone temperature and on the manifestation of tomato spotted wilt symptoms and yield of tomato

Tomato spotted wilt (TSW) disease is a serious constraint to tomato production in various regions of the world. The effect of TSW on tomato yield is largely influenced by time of infection. Early infection usually results in severe stunting of the seedling and even death of the plant. Plastic film mulches affect both the incidence of TSW, and plant growth and yield of tomato. The objective of the present study was to determine the effect of root zone temperature (RZT) as affected by plastic film mulch on the manifestation of symptoms of TSW, and growth and yield of tomato plants either artificially inoculated with tomato spotted wilt virus (TSWV) or under natural TSW infection. In artificially inoculated plants as well as in plants under natural TSW infection, vegetative top fresh weight (FW) and fruit FW both increased with the length of time after transplanting that the plants remained free from TSW symptoms. The root zone temperature was highest under black mulch (seasonal mean = 27.5 °C), followed by gray (27.0 °C), silver (25.8 °C), and white (24.8 °C) mulches. The plants grown on black mulch showed the earliest appearance of TSW symptoms, and had significantly reduced vegetative growth and fruit yields compared to plants on the other mulches. In conclusion, utilization of plastic mulches that created conditions of high RZT stress resulted in reduced plant growth and yield and predisposed the plants to earlier expression of TSW symptoms compared to plants grown at RZTs more favorable to tomato plant growth (optimal RZT = 26.1 °C [Díaz-Pérez, J.C., Batal, K.D., Granberry, D., Bertrand, D., Giddings, D., Pappu, H., 2003. Vegetative top growth and yield of tomato grown on plastic film mulches as affected by the appearance of symptoms of Tomato spotted wilt virus. HortScience 38, 395–399]). Since these plant responses to TSW under heat stress occurred in artificially inoculated plants as well as in plants under natural TSW infection, high RZTs probably affected the plants directly, independently of any possible effects on the thrips vectors.     

Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction

Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL⁻¹ of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g⁻¹ tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.     

牙鲆血清免疫球蛋白的分离纯化及部分特性分析

运用Sephacryl S-200凝胶层析和HiTrap rProtein ASepharose亲和层析2种方法对牙鲆(Paralichthys olivaceus)血清免疫球蛋白进行分离纯化,结果表明,牙鲆免疫球蛋白分布于33%~50%的硫酸铵饱和溶液中,其中45%的分离效果最好。凝胶层析和亲和层析样品均出现2个蛋白峰,用还原SDS-PAGE检测确定牙鲆免疫球蛋白存在于第2个蛋白峰中。牙鲆免疫球蛋白重链分子量约为75.4 kD,轻链分子量约为29.9 kD和28.2 kD,推测牙鲆血清免疫球蛋白的分子量为836 kD。制备了兔抗牙鲆免疫球蛋白多克隆抗体,免疫双扩散法检测多克隆抗体效价为1∶32,免疫斑点法检测多克隆抗体效价至少为1∶1 600。运用免疫印迹法(Western-bloting)检测了兔抗牙鲆免疫球蛋白多克隆抗体的特异性,实验证明该抗体与牙鲆全血清中免疫球蛋白重链、轻链反应均成阳性。     

圆斑星鲽线粒体基因组全序列结构及其进化

利用圆斑星鲽(Verasper variegatus Temminck et Schlegel)和相关鱼类的部分线粒体基因序列,设计出6对扩增引物,通过PCR扩增产物直接测序和引物行走(Primer walking)法测定圆斑星鲽线粒体基因组全序列,并对其进行结构与进化分析。圆斑星鲽线粒体基因组序列长17273 bp,其基因序列及构成都与其他硬骨鱼基本相同,包括37个基因(2个rRNA基因、22个tRNA基因和13个蛋白质编码基因)和2个非编码区(控制区和WANCY区)。在22个tRNA基因中,tRNAGln、tRNAAla、tRNAAsn、tRNACys、tRNATyr、tRNASer(第2个)、tRNAGlu和tRNAPro的编码基因位于L链上,其余则位于H链上。在13个蛋白质编码基因中,除了COⅠ基因的起始密码子为GTG外,其余均以ATG为起始密码子。蛋白质编码基因包括具有完整TAA终止密码子的ND1,COⅠ,ATP8,ND4L,ND5,而其他蛋白编码基因则具有不完全终止密码子。在L链上,ND6是仅有的一个蛋白质编码基因。圆斑星蝶的控制区包含1个终止相关序列区(ETAS)、6个中央保守序列区(CSB-A、B、C、D、E、F)和3个保守序列区(CSB-1、2、3)以及长串联重复区(Tandemly repeat sequence)。用NJ法和MP法对5个目22种鱼mtDNA的13个蛋白质编码基因的氨基酸序列进行系统分析,结果显示,圆斑星蝶与石鲽关系最近,鲽形目的鲆、鲽类与鲈形目的科(Carangidae)、鲷科(Sparidae)鱼类亲缘关系较近,而同属于鲽形目的塞内加尔鳎(Solea senegalensis)没有和任何目聚在一起,成为一个独立的分支。圆斑星蝶的基因组全序列序列已提交到GenBank,登录号为DQ403797,控制区单元型序列的GenBank登录号为DQ834444~7。     

牙鲆5个养殖群体的遗传多样性分析

利用16对微卫星分子标记对牙鲆（Paralichthys olivaceus）的5个养殖群体进行遗传多样性分析。结果表明，等位基因数为2-9个，平均等位基因数为6．0625；有效等位基因数1．2596-5．5161，平均有效等位基因数3．692；各位点的杂合度观测值（Ho）0．2200-0．8000；杂合度期望值（He）0．2061-0．8187。各群体之间无偏倚杂合度期望值从小到大依次为丹东、北戴河、威海、青岛、荣成。Kruskal-Wallis检验结果（H=0．672，df=4，P=0．955）则说明，5个群体遗传多样性差异不显著。群体间基因分化系数（Gs，）为0．0991，各群体之间存在中度遗传分化。采用UPGMA法对5个群体进行聚类，可分3类：丹东和北戴河各为一类，威海、荣成、青岛为一类，其中威海与荣成群体的分化最小。结合Hardy-Weinberg平衡，拟合度检验和遗传偏离指数（动的分析结果表明，各群体的遗传平衡状况存在很大差别。遗传变异的分析结果说明5群体遗传多样性比较丰富。     

Cloning, characterization and expression of cDNA containing major histocompatibility complex class I, IIα and IIβ genes of Japanese flounder Paralichthys olivaceus

ABSTRACT:   The nucleotide sequences of Japanese flounder Paralichthys olivaceus , major histocompatibility complex (MHC) cDNA, classical MHC class Iα, non-classical MHC class Iβ, MHC class IIα and IIβ, were determined. The domain structures and antigen binding motifs of vertebrate MHC are conserved in the Japanese flounder MHC. A phylogenetic analysis supports the classification of these genes into class I and class II MHC. Classical MHC class Iα was ubiquitously expressed, whereas the non-classical MHC class Iβ was expressed mainly in lymphoid organs, gills, intestine and stomach. The MHC classes IIα and IIβ were also ubiquitously expressed.