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1.
采用RT—PCR技术首次从孤雌生殖长角血蜱四川株克隆到P27／30基因,扩增序列全长670bp,包含完整的开放阅读框,编码201个氨基酸,预测蛋白相对分子质量为23．38ku。同源性分析表明孤雌生殖长角血蜱中国株与日本株P27／30基因同源性高达99．85％。经RT—PCR检测分析,该基因在孤雌生殖长角血蜱的卵、幼蜱、若蜱、饥饿成蜱和饱血成蜱这几个阶段均有表达。将该基因亚克隆后连接到pET32a（＋）原核表达载体,转化BL21（DEa）宿主菌,经IPTG诱导可成功进行表达。表达的目的蛋白大小为24ku左右,与预期大小一致;Western-blot显示兔抗长角血蜱全虫抗体能够识别该重组表达蛋白。 相似文献
2.
参照GenBank中长角血蜱致病性Okayama株肌钙蛋白P27/30基因的核苷酸序列,设计合成一对引物,从本实验室保藏的干净长角血蜱卵中快速提取总RNA,通过RT-PCR扩增出607 bp的肌钙蛋白基因,将其克隆于PGEM-T-Easy载体,对其进行序列分析,并推导出氨基酸序列,将这一序列与国内外已发表的长角血蜱肌钙蛋白基因进行比较分析。结果表明本实验室保藏的长角血蜱甘肃株与上述已发表的Okayama株核苷酸序列同源性为92.1%,氨基酸同源性为86%。长角血蜱甘肃株与Okayama株、四川孤雌生殖株的发育有一定差异。经RT-PCR分析表明,该基因在长角血蜱的卵、幼蜱、若蜱和饥饿成蜱这几个阶段均有表达。 相似文献
3.
《畜牧兽医学报》2010,(6)
本研究旨在阐明长角血蜱与嗜群血蜱间的亲缘关系。采用聚合酶链式反应(PCR)技术从长角血蜱和嗜群血蜱基因组DNA中扩增得到核糖体第二内部转录间隔区基因(ITS-2)片段、线粒体细胞色素氧化酶亚基Ⅰ基因(COⅠ)片段和线粒体细胞色素氧化酶亚基II基因(COⅡ)片段,并进行测序,进而分析其基因变异与系统发育。结果表明,长角血蜱和嗜群血蜱的ITS-2、COⅠ和COⅡ基因片段的大小存在差异,长角血蜱3种基因分别为361、479和647bp,嗜群血蜱基因分别为354、474和661bp。长角血蜱与嗜群血蜱间ITS-2、COⅠ和COⅡ基因的相似性分别为84.2%、89.0%和88.4%。以3种基因构建的NJ进化树中,长角血蜱和嗜群血蜱均聚类。因此,认为长角血蜱和嗜群血蜱间ITS-2、COI和COII基因间变异较小,它们是血蜱属中亲缘关系较近的2个有效种,支持传统形态学的分类地位。 相似文献
4.
本研究旨在阐明长角血蜱与嗜群血蜱间的亲缘关系.采用聚合酶链式反应(PCR)技术从长角血蜱和嗜群血蜱基因组DNA中扩增得到核糖体第二内部转录间隔区基因(ITS-2)片段、线粒体细胞色素氧化酶亚基Ⅰ基因(COI)片段和线粒体细胞色素氧化酶亚基Ⅱ基因(COⅡ)片段,并进行测序,进而分析其基因变异与系统发育.结果表明,长角血蜱和嗜群血蜱的ITS-2、COⅠ和COⅡ基因片段的大小存在差异,长角血蜱3种基因分别为361、479和647 bp,嗜群血蜱基因分别为354、474和661 bp.长角血蜱与嗜群血蜱间ITS-2、COⅠ和COⅡ基因的相似性分别为84.2%、89.0%和88.4%.以3种基因构建的NJ进化树中,长角血蜱和嗜群血蜱均聚类.因此,认为长角血蜱和嗜群血蜱间ITS-2、COⅠ和COⅡ基因间变异较小,它们是血蜱属中亲缘关系较近的2个有效种,支持传统形态学的分类地位. 相似文献
5.
根据已发表的长角血蜱16SrRNA序列及斑点热群立克次体外膜蛋白A（OmpA）基因序列设计2对特异性引物,对唐山地区采集的长角血蜱进行PCR检测,并对阳性样本进行测序和序列分析,抽检样本建立分子系统进化树。结果表明,在315份蜱DNA样本中检测出25份阳性样本,阳性率为7.94%;序列分析结果显示唐山地区长角血蜱携带立克次体同处于一个分支,与日本株立克次体同源性最高（93.30%）,其次是福建株立克次体（92.11%）,黑龙江立克次体绥芬株（90.45%）、虎林株（90.42%）。结论得出唐山地区蜱传斑点热感染较严重,分子进化分析结果显示唐山地区蜱传斑点热群立克次体可能为一新种。 相似文献
6.
本试验从镰形扇头蜱半饱血雌蜱唾液腺cDNA文库的EST中筛选了1个含polyA尾的基因序列,经5LRACE方法得到该基因全长序列.经同源性比较,该基因预测的氨基酸序列与长角血蜱肌钙蛋白I(GI14041807)的同源性为84.47%,肌动蛋白结合位点位于147-167氨基酸处,且与长角血蜱肌钙蛋白I肌动蛋白结合位点完全相同,表明该基因是镰形扇头蜱肌钙蛋白I基因.以镰形扇头蜱基因组DNA为模板扩增到编码肌钙蛋白I的基因组序列.序列分析表明该序列不含内含子.RT-PCR分析表明该基因在镰形扇头蜱的卵及其幼蜱、若蜱、成蜱的壳、唾液腺和肠均有表达. 相似文献
7.
牛的巴贝斯虫18S rRNA基因序列比较研究 总被引：10，自引：2，他引：10
对中国已报道的8株牛的巴贝斯虫（包括1株牛巴贝斯虫、1株双芽巴贝斯虫、1株大巴贝斯虫、3株卵形巴贝斯虫和2株巴贝斯虫未定种）的18S rRNA基因序列进行了测定与比较。自感染动物的血液中纯化虫体，提取基因组DNA，PCR扩增靶基因，然后将其连接到pGEM—T Easy载体上，进行克隆测序。研究结果显示：牛的巴贝斯虫18S rRNA基因大小在1653～1699bp之间；用所测得的和自GenBank下载的各种动物的巴贝斯虫18S rRNA基因序列构建了系统发生树，发现由刻点血蜱传播的大巴贝斯虫伊犁株与由长角血蜱传播的3株卵形巴贝斯虫存在明显差别，应属于2个独立种；由小亚璃眼蜱传播的牛巴贝斯虫未定种不同于目前已报道的任何种类，在中国应为一个新种。因而，中国存在5种牛的巴贝斯虫，即：牛巴贝斯虫，双芽巴贝斯虫、大巴贝斯虫，卵形巴贝斯虫和巴贝斯虫未定种。 相似文献
8.
为了对陕西省和辽宁省部分地区羊源蜱类及其携带病原体的流行情况进行分析,本试验采集了288份羊源蜱样本,采用形态学观察鉴定蜱种;采用分子生物学方法扩增巴贝斯虫18S rRNA、伯氏疏螺旋体16S rRNA、斑点热群立克次体ompA和无形体16S rRNA的基因序列,并结合核苷酸序列分析,确定所收集蜱样本中各类病原体的流行情况。结果显示,经鉴定288份羊源蜱样本均为长角血蜱。陕西省144份长角血蜱中携带巴贝斯虫、斑点热群立克次体和无形体,感染率分别为0.69%、21.53%和77.78%,且巴贝斯虫和斑点热群立克次体均与无形体复合感染;其中长角血蜱中携带Babesia microti、Candidatus Rickettsia longicornis、Uncultured Rickettsia sp. QH-122、Rickettsia endosymbiont of Haemaphysalis longicornis、Uncultured Anaplasma sp. ZJ06/2009、Anaplasma marginale、Anaplasma capra共7种病原体。辽宁省144份长角... 相似文献
9.
《动物医学进展》2015,(12)
为了解山东省沂源县境内硬蜱携带无形体的流行及菌株变异情况,于2015年5月~7月从该地区山羊体表共分离到54份126只不同生活史阶段的长角血蜱,分组研磨提取DNA后,利用套式PCR扩增无形体的16SrRNA(ribosomal RNA)基因片段并测序及进行序列的遗传进化分析。结果显示,54份样品PCR阳性扩增率为48.1%;序列分析表明,该地区无形体存在6个变异株,分为4类,其中3类与该地区以往报道的嗜吞噬细胞无形体序列保持密切的遗传关系,还有1类与日本长角血蜱体内检出的牛无形体序列遗传关系较近。由此可见,山东沂源地区长角血蜱存在较高的嗜吞噬细胞无形体感染率,并且变异株较多;应该加强该地区对人粒细胞无形体病的宣传力度,防止该病大规模暴发。 相似文献
10.
镰形扇头蜱肌钙蛋白Ⅱ基因的克隆及其分布 总被引：1，自引：0，他引：1
本试验从镰形扇头蜱半饱血雌蜱唾液腺cDNA文库的EST中筛选了1个含polyA尾的基因序列,经5′-RACE方法得到该基因全长序列。经同源性比较,该基因预测的氨基酸序列与长角血蜱肌钙蛋白（GI：14041807）的同源性为84.47%,肌动蛋白结合位点位于147～167氨基酸处,且与长角血蜱肌钙蛋白肌动蛋白结合位点完全相同,表明该基因是镰形扇头蜱肌钙蛋白基因。以镰形扇头蜱基因组DNA为模板扩增到编码肌钙蛋白的基因组序列,序列分析表明该序列不含内含子。RT-PCR分析表明该基因在镰形扇头蜱的卵及其幼蜱、若蜱、成蜱的壳、唾液腺和肠均有表达。 相似文献
11.
The reproductive mechanism of Haemaphysalis longicornis is quite different from many other animal species. In this article, several characteristics of parthenogenetic and bisexual populations of H. longicornis were analyzed, including some important micro-structures, synchronized life cycle feature and sequences of mitochondrial 16S rRNA gene. The results suggested even though many observations of the two populations were similar to each other, some important differences also existed. The genital apron of parthenogenetic females was wider than that of bisexual females. Parthenogenetic individuals (except engorged females) were significantly larger in weight than bisexual individuals (p<0.01; unfed nymph p<0.05); the difference of nymphal premoulting, female feeding and preoviposition, and egg incubation periods between the two populations were significant (p<0.01); hatch percentage of parthenogenesis was lower than that of bisexual population (69% and 73%, respectively); parthenogenetic individuals had a slightly longer development cycle than bisexual individuals (134 and 129days, respectively). Hybridization attempts failed between them. Comparing to the two strains of bisexual H. longicornis, parthenogenetic strain of H. longicornis inserted two nucleotides of thymine, though the genetic distance of 16S rDNA between the bisexual and parthenogenetic populations was 0. In order to clarify the relationships of the two reproductive populations of H. longicornis, the sequences of 16S rDNA of four strains of two other species were also analyzed. The divergence between Hebei and Xinjiang strains of Rhipicephalus sanguineus was 0, and the sequences were totally identical between them. The divergence between the two subspecies of Hyalomma asiaticum was 0.01. The results indicated that the relationship between bisexual and parthenogenetic H. longicornis was certainly closer than that between subspecies, but farther than that between the same reproductive populations of conspecies. 相似文献
12.
N Sasaki K Takahashi S Kawamoto T Kurosawa M Sonoda S Kawai 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(3):495-501
The parthenogenetic Haemaphysalis longicornis larvae engorged on cattle naturally infected with Theileria sergenti were reared at 24 degrees C. The resultant nymphal ticks were incubated at 37 degrees C to clear the effect of incubation on the development and maturation of sporozoites. The sporozoites in the salivary glands of the nymphal ticks exposed to 37 degrees C for 16 days were observed by the methyl green pyronin staining method. The ticks exposed to 37 degrees C were ground up in a mortar and the supernatant of the tick suspension in PBS was inoculated into cattle. The cattle showed parasitemia and specific antibody response 18 days after inoculation. Consequently, the parasites in the tick salivary glands became infective to cattle by incubating infected. H. longicornis nymphs at 37 degrees C. 相似文献
13.
Various species of ixodid ticks, attached to domestic cats in Japan, were identified in spring (April to June) and autumn (September to November). In the spring, a total of 282 ticks, including 61 larvae, 70 nymphs, 127 females and 24 males were collected from 126 cats. Of these, 264 were identified up to the species level. In the spring, Haemaphysalis longicornis was the most frequently (39.7%, 50/126) found tick species on feline hosts, followed by Ixodes ovatus (35.0%, 44/126), Ixodes nipponensis (15.9%, 20/126) and Haemaphysalis flava (9.5%, 12/126). Small numbers of Haemaphysalis megaspinosa, Haemaphysalis japonica, Ixodes persulcatus, Ixodes granulatus and Amblyomma testudinarium were also recovered. H. longicornis was the most frequently found tick species on cats around riversides or river basins, while I. ovatus and I. nipponensis were more frequently found on cats kept near woodland or related areas. I. nipponensis was more frequently found on castrated males. No major statistical differences in the frequency of tick attachment among sex, age or hair length for the three major tick species were found. Of 205 ticks including 173 (84.4%) larvae, 27 (13.2%) nymphs, 4 (2.0%) females and 1 (0.5%) male recovered from 62 cats in autumn, only 32 (15.6%) were identified. Most of the larvae were fully- or partly-engorged Haemaphysalis spp., and it was difficult to identify them further by morphological characterization. 相似文献
14.
Andrea Egizi Leslie Bulaga‐Seraphin Erika Alt Waheed I. Bajwa Joshua Bernick Matthew Bickerton Scott R. Campbell Neeta Connally Kandai Doi Richard C. Falco David N. Gaines Telleasha L. Greay Vanessa L. Harper Allen C.G. Heath Ju Jiang Terry A. Klein Lauren Maestas Thomas N. Mather James L. Occi Charlotte L. Oskam Jennifer Pendleton Marissa Teator Alec T. Thompson Danielle M. Tufts Rika Umemiya‐Shirafuji Meredith C. VanAcker Michael J. Yabsley Dina M. Fonseca 《Zoonoses and public health》2020,67(6):637-650
Established populations of Asian longhorned ticks (ALT), Haemaphysalis longicornis, were first identified in the United States (US) in 2017 by sequencing the mitochondrial cytochrome c oxidase subunit I (cox1) ‘barcoding’ locus followed by morphological confirmation. Subsequent investigations detected ALT infestations in 12, mostly eastern, US states. To gain information on the origin and spread of US ALT, we (1) sequenced cox1 from ALT populations across 9 US states and (2) obtained cox1 sequences from potential source populations [China, Japan and Republic of Korea (ROK) as well as Australia, New Zealand and the Kingdom of Tonga (KOT)] both by sequencing and by downloading publicly available sequences in NCBI GenBank. Additionally, we conducted epidemiological investigations of properties near its initial detection locale in Hunterdon County, NJ, as well as a broader risk analysis for importation of ectoparasites into the area. In eastern Asian populations (China/Japan/ROK), we detected 35 cox1 haplotypes that neatly clustered into two clades with known bisexual versus parthenogenetic phenotypes. In Australia/New Zealand/KOT, we detected 10 cox1 haplotypes all falling within the parthenogenetic cluster. In the United States, we detected three differentially distributed cox1 haplotypes from the parthenogenetic cluster, supporting phenotypic evidence that US ALT are parthenogenetic. While none of the source populations examined had all three US cox1 haplotypes, a phylogeographic network analysis supports a northeast Asian source for the US populations. Within the United States, epidemiological investigations indicate ALT can be moved long distances by human transport of animals, such as horses and dogs, with smaller scale movements on wildlife. These results have relevant implications for efforts aimed at minimizing the spread of ALT in the United States and preventing additional exotic tick introductions. 相似文献
15.
Tabara K Kawabata H Arai S Itagaki A Yamauchi T Katayama T Fujita H Takada N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(4):507-510
Endemic spotted fever group rickettsiosis was reported in Shimane Prefecture, Japan. From an analysis of 14 clinical cases found in the endemic area, the infectious agent of spotted fever group rickettsiosis was identified as Rickettsia japonica. In this study, we also found that Rickettsia japonica was highly infected with the vector tick, Haemaphysalis longicornis, in the endemic area. These findings suggest that the high incidence of rickettsiosis in Shimane Prefecture can be explained by the high prevalence of Rickettsia japonica among Haemaphysalis longicornis ticks. 相似文献
16.
每隔2周用长角血蜱成蜱定量感染家兔,成功构建了具有不同抵抗力的家兔免疫模型。ELISA检测结果表明,家兔血清中的抗体效价从初次叮咬后第3周开始呈阳性,随着叮咬次数的增加抗体水平逐渐上升,第11周时达到高峰。具有一定抵抗力的家兔对长角血蜱成蜱的吸血和生长发育影响显著,其抗体水平与长角血蜱成蜱在兔体的吸血周期、死亡率成正相关,与雌蜱的饱血脱落率、饱血体重成负相关。结果进一步证明,蜱的唾液腺是其主要的免疫器官之一。家兔抗体水平的持续期足以使长角血蜱完成一个世代,可以满足试验需求,是抗蜱免疫研究理想的阳性对照模型。同时,血清中特异性抗体的效价也可以作为判断动物对长角血蜱抵抗力强弱的重要指标之一。 相似文献
17.
Miyoshi T Tsuji N Islam MK Kamio T Fujisaki K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(11):1471-1473
RNA interference (RNAi) has been recently exploited to determine gene function by degrading specific mRNAs in several eukaryotic organisms. We constructed a double stranded RNA (dsRNA) from a previously cloned Haemaphysalis longicornis serine proteinase (HlSP) gene to test the importance of the function of the HlSP gene product during blood-feeding. Growth of unfed ticks treated with HlSP dsRNA was significantly inhibited compared to that of PBS-treated ticks. This inhibition was supported by the level of HlSP mRNA. HlSP may play a crucial role for blood-feeding in these ticks. This is the first report on gene silencing of a functional serine proteinase in hard ticks. 相似文献
18.
对采自中国东洋区贵州省、云南省、浙江省、江苏省的“二棘血蜱”和采自中国古北区吉林省、河南省、河北省、辽宁省的长角血蜱的生活周期及其幼虫、若虫和成虫的形态特征作了比较观察。结果,与 Hoogstraal提出的二棘血?的鉴别点相对照,除相当一部分“二棘血蜱”成虫盾板上的刻点比较细这一点符合Hoogtsraal所提出的二棘血蜱的待征外,所观察的“二棘血蜱”的幼虫,若虫和成虫反比长角血蜱大;二者若虫的齿式均为3/3,须肢第3节背后缘中央均无突起,二者成虫的齿式均为5/5,盾板上的刻点均稠密、分布均匀;通过扫描电镜观察,二者哈氏器结构也无明显的区别。因此认为,采自中国东洋区的所谓“二棘血蜱”实际是长角血蜱。然而分布于中国东洋区和古北区的长角血蜱间,形态上存有一定程度变异,具体表现在个体大小、幼虫肩突尖端间距离、若虫和成虫须肢第2节外角的突出程度、成虫盾板上刻点的粗细以及若虫哈氏器远端缝孔的有无等方面。 相似文献
19.
为深入研究肠道细菌在蜱的生理学和传播病原上的作用,本研究进行了长角血蜱肠道细菌的分离鉴定。按照细菌传统分离方法并结合16S rDNA测序鉴定,获得7个分离株,这些菌株分别属于显核菌属(Caryophanon sp.)、考克氏菌属(Kocuria sp.)、葡萄球菌属(Staphylococcus sp.)、微杆菌属(Microbacterium sp.)和短杆菌属(Brevibacterium sp.)5个菌属。 相似文献