狐狸体表犬啮毛虱的形态特征观察和分子鉴定

本研究旨在利用全证据法对采自湖南省长沙生态动物园狐狸体表的2只寄生虱进行准确鉴定。首先通过显微镜观察寄生虱的形态特征，初步鉴定为毛虱。然后分别提取2只寄生虱的DNA样本，再以PCR扩增寄生虱线粒体细胞色素c氧化酶I亚基（cox1）基因的部分序列，进行序列分析，并利用Mega 11程序最大似然法（ML）构建系统发育树，从分子水平进一步对寄生虱进行鉴定。PCR扩增结果显示，两个样本条带大小与预期一致，均约400 bp；序列分析显示两个样本cox1序列与GenBank收录的犬啮毛虱（Trichodectes canis，基因登录号MH001214）相似性均为99.0%；系统发育分析显示，两个样本均与犬啮毛虱位于同一分支。结果表明，采自狐狸体表的寄生虱为犬啮毛虱。本研究为湖南省不同宿主犬啮毛虱流行病学调查提供了新的分子学证据。     

新疆鹰羽虱形态鉴定初报

本文利用体视显微镜以及扫描电子显微镜(SEM),对采自新疆鹰共78只外寄生虫进行了亚微形态观察。结果显示:其光学显微结构为无翅、足三对、触角两对,细长伸出于头外,头部大于胸且后颊突出呈角状,腹部长度大于头胸之和;超微结构特点为触角分5节(第一节最大,依次减小),咀嚼式口器呈卵圆形,气孔与气门板未见典型特点,初步鉴定为长角羽虱科(philopteridae)的羽虱,可为我国野生禽类羽虱的分类提供参考。     

爪哇花蜱的形态学和分子标记特征

为了便于识别和鉴定爪哇花蜱,本试验从湖南省长沙市南郊公园穿山甲体表采集了爪哇花蜱,并借助体视显微镜超景深系统观察雌、雄成蜱的形态结构,通过分子生物学技术测定了COX1和16S rDNA基因序列。结果显示,雌性成蜱假头基呈矩形;孔区小而深;须肢前宽后窄,第2节长;盾板暗赤褐色,无珐琅斑,心脏形;眼小而扁平,不明显,仅见一痕迹;肛沟位于肛门后;气门板呈逗点形。雄性成蜱假头基、须肢、眼、肛沟和气门板特征与雌蜱相一致;盾板特点为呈宽卵圆形,无珐琅斑,周缘暗赤褐色,中部稍浅,呈黄褐色。爪哇花蜱COX1基因序列与国外花蜱属蜱种Amblyomma fimbriatum、Amblyomma triguttatum等的COX1基因序列相似度均为83%;爪哇花蜱16S rDNA基因序列与嗜龟花蜱、龟形花蜱等花蜱的16S rDNA基因序列相似度为85%～88%,相似度均较低。结果表明:爪哇花蜱具有显著、易辨识、易鉴定的形态学和分子标记特征。     

鼷鼠肉螨的形态学观察和分子鉴定

本试验旨在对患病小鼠感染的螨进行鉴别诊断，首先，使用透明胶纸法和被毛采集法在显微镜下进行形态学观察鉴定，初步确定螨的种类；随后，选定螨18S rRNA基因，建立聚合酶链式反应(PCR)快速检测方法，并进行测序比对分析，从分子水平上进一步确认螨的种类。形态学观察结果显示，小鼠感染的螨呈白色，长椭圆形，中央稍宽，前后两端稍窄，有4对足；在螨体后3对足之间有特征性的突起；尾端的尾乳突有2根特征性的后端毛，初步鉴定为鼷鼠肉螨。PCR扩增虫体18S rRNA基因和序列比对结果显示，分离螨与鼷鼠肉螨的18S rRNA基因序列核苷酸相似性达98.68%。直接镜检结合PCR检测结果，表明患病小鼠感染了鼷鼠肉螨。     

鲤鱼双线绦虫裂头蚴的分子鉴定和系统发育关系分析

为了鉴定天津市某养殖场鲤鱼腹腔内寄生的“面条样”虫体，本试验通过PCR方法扩增虫体的18S rRNA、COⅠ和COB基因序列，并进行分子鉴定和基因测序，分析3种基因的同源性、分子进化和系统发育关系。结果显示，寄生于鲤鱼体内的虫体的18S rRNA、COⅠ和COB基因序列分别与双线绦虫分离株MN204040.1(中国)、MN219466.1(中国)和EU241209.1(法国)的核苷酸序列同源性较高，为99.45%、98.74%和99.26%。基于18S rRNA、COⅠ和COB基因序列构建的系统发育树显示，本试验分离的绦虫裂头蚴与GenBank数据库中已鉴定的双线绦虫聚在同一分支上，且与双线绦虫中国武汉分离株的种属亲缘关系较近。本试验初步阐明获得的天津市双线绦虫分离株与其他种属之间的系统发育关系，为双线绦虫的分子鉴定和系统发育关系分析提供了数据支持。     

鸡快慢羽分子鉴定方法的建立及其应用

利用找到的鸡快羽和慢羽基因,采用双重PCR方法建立了一种高效的鸡快、慢羽分子鉴定方法,在快慢羽自别雌雄鸡群体当中验证表明准确率达100%。采用该方法和经典表型鉴定方法对同一群鸡进行快慢羽判定,结果发现,除表型鉴定为微长型的个体与分子鉴定结果符合率为94.1%外,其它羽型符合率均达100%。对该鸡群快、慢羽个体早期生长速度比较研究发现,除出壳重差异不显著、第1周差异显著(P0.05),在其后各周,快、慢羽个体间体重差异均极显著(P0.01)。     

虱螨净对鸡羽虱的治疗效果观察

本实验对伊犁河谷地区白色和黑色羽虱采用传统的杀虫药—敌百虫和新型驱虫药—虱螨净分别对自然感染羽虱的鸡进行治疗,结果表明传统方法治疗效果差,易复发。使用虱螨净进行治疗后治愈率可达到100%,且2个月后无复发现象。     

鸿雁和东方白鹳体表虱子的形态鉴定

对采自哈尔滨北方森林动物园鸿雁和东方白鹳体表的2种虱子通过形态结构观察、显微测量,并进行形态学鉴定。结果表明,寄生在鸿雁体表的虱子为鹅啮羽虱(Esthiopterum anseris),寄生在东方白鹳体表的虱子为鸭羽虱(Trinoton quequedulae)。     

鹅羽虱的防治

鹅的羽虱严重影响羽绒和裘皮的质量，同时生产力降低，生长受阻。笔者就鹅羽虱防治问题谈一点体会。     

山羊毛虱的形态结构和分子标志研究

为了解山羊毛虱形态结构和分子标志,用电子显微镜和三维立体显微镜观察了采自山羊的山羊毛虱形态结构,测定了其18SrDNA和cox1基因序列,并基于比对,揭示其序列特点。形态观察发现,触角3节,第1节短粗,第2、3节细长;中胸气门1对,位于前胸两侧;腹部有气门6对;跗节3节。序列分析显示,本样虱、山羊毛虱、具边毛虱和牛毛虱四者的18SrDNA基因序列高度相似,不适于做种间鉴定。cox1序列极不保守。     

山羊腔阔盘吸虫的形态学观察和分子鉴定

为了从形态学和分子水平对山羊腔阔盘吸虫(Eurytrema coelomaticum)的种类鉴定提供依据,对采集自广东佛山发病山羊胰脏的阔盘吸虫在形态学观察的基础上,抽提虫体基因组DNA,通过PCR方法对虫体核糖体18SrRNA基因和线粒体cox1基因进行扩增,将产物与pMD18-T载体连接,转化到大肠埃希菌DH5α感受态细胞进行克隆、测序和序列分析。结果获得虫体18SrRNA基因的大小为1 922bp,线粒体cox 1基因大小为444bp。序列分析显示,获得的18SrRNA基因与GenBank收录的腔阔盘吸虫(E.coelomaticum,登录号为DQ401035)同源性高达99%;线粒体cox1基因与GenBank收录的胰阔盘吸虫(E.pancreaticum,登录号为KC535544)的同源性仅为90%。研究结果从分子水平证明采集自广东佛山山羊体的阔盘吸虫为腔阔盘吸虫(E.coelomaticum),对山羊阔盘吸虫病的防控有指导意义。     

肝片吸虫蚴虫的形态学及分子学鉴定

为了找到青海地区肝片吸虫(Fasciola hepatica)的主要中间宿主,并对椎实螺(Lymnaeidae snails)体内的蚴虫进行虫种鉴定。对青海部分地区的椎实螺进行压片镜检,结果发现椭圆萝卜螺(Radix swinhoei)内无肝片吸虫蚴虫感染。狭萝卜螺(Radix lagotis)体内发现雷蚴,雷蚴为肝片吸虫的蚴虫,但是感染率较低为10%。青海萝卜螺(Radix cucnnorica)体内发现了雷蚴和尾蚴,并在养殖青海萝卜螺的水中青菜叶子上收集到囊蚴,经初步鉴定均为肝片吸虫的蚴虫,感染率较高为41.89%。主要中间宿主螺为青海萝卜螺。提取蚴虫体内DNA,通过PCR扩增18S rRNA片段并测序。将测序用DNAMAN软件进行比对后发现,与Gen Bank中发布的大片吸虫(Fasciola gigantica)和肝片吸虫的18S rRNA的序列进行比对其相似度分别为99.12%和92.03%。可以进一步确定所收集的蚴虫为肝片吸虫的蚴虫。     

Morphological and Molecular Biology Identification of Ornithodoros lahorensis

This study was to establish a method for the identification of Ornithodoros lahorensis with molecular biology. The samples of Ornithodoros lahorensis were collected from sheep in Xinjiang region and the morphological characters were screened by stereo microscopy and electronic microscopy. Then the 16S rDNA,18S rDNA and cytochrome oxidase Ⅰ(COⅠ) of samples were amplified by PCR and subsequently sequenced. Then the analysis of genetic divergences and Neighbor-Joining (NJ) phylogenetic tree were carried out based on the sequences acquired from Ornithodoros lahorensis,combined with the reference sequences of GenBank database. In this study,the comprehensive and clear micromorphological traits of ovoid scutum,round fovea of Ornithodoros lahorensis were provided. Besides,the DNA sequences of Ornithodoros lahorensis were assembled together with sequences of Argasidae and form monophyletic clades in the 16S rDNA and COⅠ based Neighbor-Joining trees. According to the intraspecific K2P genetic distances of Ornithodoros lahorensis (0~0.3%),we determined that the molecular biology method based on 16S rDNA and COⅠ could rapidly and accurately identify the species of Ornithodoros lahorensis.     

拉合尔钝缘蜱的形态学及分子生物学鉴定

试验旨在建立拉合尔钝缘蜱的分子生物学鉴定方法。从新疆地区绵羊体表采集寄生蜱,借助体视显微镜和电子显微镜对其进行形态学特征的准确鉴别,初步筛选出疑似拉合尔钝缘蜱。经PCR扩增、测序获得其18S rDNA、16S rDNA及线粒体色素氧化酶亚基Ⅰ基因（cytochrome oxidase Ⅰ,COⅠ）序列,结合GenBank数据库中参考序列,比对分析同源性并构建系统进化树,进行遗传距离评估。本研究全面、清晰地揭示了拉合尔钝缘蜱卵圆形背板、背部圆形盘窝等显微形态特征的细节。同时,在基于16S rDNA与COⅠ序列的邻位相接系统进化树中,拉合尔钝缘蜱分布于两大进化枝之一的软蜱科,同种聚类良好,且拉合尔钝缘蜱种内K2P遗传距离为0~0.3%,本研究方法可快速、准确鉴定拉合尔钝缘蜱物种。     

Control of the cattle louse Bovicola bovis with the fungal pathogen Metarhizium anisopliae

The effects of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) were evaluated against the common louse parasite of cattle, Bovicola bovis (Piaget) (Trichodectidae: Ischnocera). Two different concentrations and formulations of conidial suspensions were applied to contained populations of adult female lice. In vitro, lice immersed in suspensions of M. anisopliae formulated in 0.03% Tween 80 developed infections; at the highest concentration (1x10(8)conidia ml-1) a mean of 71% (+/-11.52%, 95% C.I.) of lice became infected. Lice exposed to the Tween 80 only in vitro, showed high levels of survival and zero infection. In vivo, fungal conidia were applied to louse populations contained in 7 cm diameter circular arenas glued to the backs of Holstein cattle, maintained in controlled climate conditions. Conidia were formulated in either Tween 80 or silicone oil. The treatment with M. anisopliae resulted in high levels of infection and there was no overall difference between the two formulations in the number of infections observed. At the highest concentration (1x10(8)conidia ml-1) a mean of 73% (+/-15.57%, 95% C.I.) lice became infected. It is concluded that the strategic seasonal use of a fungal pathogen on cattle, applied in early winter, may be of value in suppressing the winter increase in abundance, preventing the population increasing to clinically significant levels.     

Geographical variation of the optimum temperature for in vitro rearing of Bovicola bovis

The indigenous biting louse of cattle, Bovicola bovis, has been satisfactorily bred in in vitro culture in Ireland. The optimum temperature was found to be 33 ± 0.5°C at a relative humidity of 75%. This temperature range is at variance with the optimun temperature ranges recorded by two authors in the United States. It is possible that geographical genetic plasticity occurs in Bovicola bovis, thus accounting for these differences.     

Persistent efficacy of doramectin topical solution against induced infestations of Bovicola bovis and Solenopotes capillatus.

A study was conducted in two locations, Wyoming and Wisconsin, USA, to evaluate the persistent efficacy of doramectin topical solution at a dose rate of 500 microg/kg body weight against artificially induced infestations of Bovicola bovis and Solenopotes capillatus on cattle. At each location, lice-free beef calves were individually housed and randomly allocated to treatment. Both B. bovis and S. capillatus were transferred from untreated donor animals to doramectin-treated cattle at the end of 35, 63, 91 or 126 day post-treatment periods. Cattle treated with a saline pour-on served as the control. Based on the geometric means of lice counts 2 weeks following transfer, the persistent efficacy of a single treatment with doramectin topical solution against induced infestations of B. bovis was 100.0, 100.0, 99.5, and 100.0% at post-treatment days of 35, 63, 91, and 126, respectively. Persistent efficacy against induced infestations of S. capillatus, for the same intervals, were 100.0, 94.9, 86.3, and 74.9%.     

广州桑园桑树根结线虫的形态学与分子鉴定

桑树根结线虫病是华南蚕区危害严重的土传病害之一.针对华南蚕区桑园根结线虫危害普遍发生的现状,对采自华南蚕区的桑园桑根结线虫病样进行了病原线虫的分离与形态学观察,并对其rDNA、CO Ⅰ基因分别进行了测序和系统发育分析,同时在基于桑根结线虫特异分子检测基础上进行了线粒体全基因组的测序比较.结果 表明,广州桑园桑根结线虫病样分离的根结线虫具有典型的象耳豆根结线虫(Meloidogyne enterolobii)的形态特征;其rDNA与CO Ⅰ基因序列与已报道的象耳豆根结线虫序列一致性均高达99％以上;rDNA与CO Ⅰ的进化树分析发现其与象耳豆根结线虫同源序列均聚在同一枝;PCR的特异性检测结果表明,只有象耳豆根结线虫的特异性引物扩增,获得236 bp的目的 条带;线粒体全基因组序列全长为17080 bp(GenBank收录号:MW186714),与GenBank唯一报道的象耳豆根结线虫M.enterolobii(KP202351.1)一致性达99.7％.基于上述的生物学观察和分子生物学研究结果,认为广州桑园的桑根结线虫病病原为象耳豆根结线虫M.enterolobii,这为桑树桑根结线虫病的发生和防控提供了实验参考.