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1.
2008年7月末,我国青岛即墨地区患病的虎皮鹦鹉幼雏出现体重下降、脱羽和羽毛变形萎缩、鸟喙变形及胸腺结构变异等症状,怀疑为鹦鹉喙羽病(PBFD)。利用PCR对濒死鹦鹉进行鹦鹉喙羽病病毒(PBFDV)的检测,并对扩增的C1基因进行了测序与分析。结果表明,有两只检测为PBFDV阳性,B last分析发现,QD-CN08株与GenBank中已发表的PBFDV分离株C1基因同源性为82%~93%,进化树分析表明与日本的毒株有比较近的亲缘关系。结合临床症状、流行病学特征和实验室诊断,确诊为鹦鹉喙羽病,此病为中国大陆首次报道。 相似文献
2.
《中国兽医学报》2018,(12)
为了解2015―2017年福建省新发猪圆环病毒3型(PCV3)的流行现状、分子生物学特征和遗传演化规律,收集福建省不同地区186个猪场的267份疑似PCV3感染猪的组织病料进行分子流行病学调查,并对其Cap基因进行了遗传变异分析。结果显示,福建地区PCV3猪场总阳性率为30.11%,疑似样品总阳性率为21.72%,且呈逐年增高的趋势;17株PCV3福建分离株与其他17株国内外参考毒株Cap基因核苷酸序列及氨基酸序列同源性较高,分别为96.0%～100.0%和96.3%～100.0%,表明这些毒株可能有相同的进化来源;氨基酸多序列比对发现,Cap基因氨基酸序列最有特征性的是第168位(R→K)和173位(L→F)的氨基酸替换,该位点的突变可能是该地区流行的PCV3毒株的一个重要分子特征;同时,抗原性分析发现第173位(L→F)的突变还位于Cap蛋白的抗原表位区内。遗传进化树分析结果表明,17株PCV3福建分离株中有2株属于3a亚型,其余15株属于3b亚型,表明当前福建地区PCV3流行毒株主要为3b亚型。 相似文献
3.
鹦鹉喙羽症病毒(PBFDV)是圆环病毒科圆环病毒属的成员,可引起多个品种鹦鹉的急性死亡或者羽毛脱落和喙变形。病毒基因组中的ORF1和ORF2分别编码病毒复制相关蛋白(Rep protein)和衣壳蛋白(Cap protein)。Cap蛋白是圆环病毒的主要结构蛋白,也是其主要的免疫保护性蛋白。本试验应用PCR技术选择性扩增PBFDV Cap蛋白抗原表位集中区域的基因片段,并将其克隆入原核表达载体pGEX-6p-1中,诱导表达截短的Cap蛋白。SDS-PAGE及质谱鉴定结果显示,融合表达的CapT蛋白约45 kDa,切胶纯化后的蛋白浓度约1. 6 mg/mL。该Cap蛋白的成功获得为PBFDV治疗用阳性血清制备及亚单位疫苗的开发奠定了必要的物质基础。 相似文献
4.
鹦鹉喙羽病(PBFD)是鹦鹉目前最常见的疾病,对鹦鹉养殖业危害极其严重。根据鹦鹉喙羽病毒(PBFDV)基因片段的克隆和序列分析,设计合成1对特异性引物,以CP基因为模板,经PCR扩增获得830 bp的核苷酸DNA,并用DIG标记DNA,制备用于检测PBFDV的特异性核酸探针。用该核酸探针对疑似感染PBFDV的鹦鹉病料进行斑点杂交检测,并对鉴定为阳性的PBFDV进行全基因组扩增和测序分析。结果显示,利用PCR结合斑点杂交技术检测PBFDV,特异性强、敏感度高,具有可重复性。鉴定为阳性的2株PBFDV全基因组序列之间同源性为100%,与已报道序列的同源性为81.5%～98.9%。本研究为我国开展PBFDV感染的分子流行病学调查和临床诊断提供了一种敏感、特异的检测方法。 相似文献
5.
为了弄清云南省猪圆环病毒2型(PCV-2)的流行和基因变异情况,试验采用PCR鉴定、PCV-2全基因扩增及克隆、PCV-2蛋白氨基酸比对与遗传进化分析、PCV-2 Cap蛋白氨基酸比对分析、PCV-2 Cap蛋白抗原指数预测分析等方法进行研究。结果表明,从43份样品中检测出5份阳性样品,其阳性率为11.63%;成功获得5个大小为1 768 bp的PCV-2全基因序列,分别将其命名为YN PCV2-1、YN PCV2-2、YN PCV2-3、YN PCV2-4、YN PCV2-5;获得的5株毒株与国内外毒株氨基酸之间存在差异,同源性在66.8%～99.1%之间;5株PCV-2 ORF2基因编码的Cap蛋白氨基酸与5株疫苗株也存在多处氨基酸变异;其中YN PCV2-4 Cap蛋白抗原指数与5株疫苗株的Cap蛋白抗原指数差异性不大,而YN PCV2-1、YN PCV2-2、YN PCV2-3、YN PCV2-5的Cap蛋白抗原指数与5株疫苗株的Cap蛋白抗原指数差异性较大,在5～10,25,40～50,70,172～176,210～223区域的抗原指数均不同于疫苗株。说明云南省流行的猪圆环病... 相似文献
6.
2018年山东省某养殖场发生大量幼龄鹦鹉死亡事件,疑似为病毒感染。为探寻病因,开展了该场及省内另外两个鹦鹉养殖场的流行病学调查。利用PCR技术对临床样品中提取的DNA或RNA进行检测,结果发现新城疫与禽流感病毒均呈阴性,而禽多瘤病毒(APV1)与鹦鹉喙羽病病毒(PBFDV)呈现为强阳性,由此推断此3个鹦鹉养殖场存在APV1和PBFDV感染,平均病毒检出率分别为67.9%与71.4%,共感染检出率率为58.0%。对阳性样品进化全基因分析发现:区域内的PBFDV流行毒株同源性高,与该地区早期报道的毒株亲缘关系较为接近;APV1的VP1基因同源性较高,与欧洲分离毒株亲缘关系较为接近,说明我国流行的APV1或来源于进口鹦鹉。本研究警示,需要加强鹦鹉疾病防控,严格鹦鹉进出口检疫,并制定和健全标准的鹦鹉病检疫程序。 相似文献
7.
本文旨在克隆禽波氏杆菌外膜蛋白OmpA的编码基因,并预测OmpA蛋白二级结构和B细胞抗原表位,从而探讨禽波氏杆菌外膜蛋白OmpA在免疫保护中所起的作用。作者对禽波氏杆菌的外膜蛋白ompA基因进行PCR扩增、克隆及序列测定。应用生物信息学相关软件和方法,对禽波氏杆菌OmpA蛋白的二级结构和B细胞抗原表位进行预测。禽波氏杆菌ompA基因全长597bp,编码199个氨基酸。二级结构以无规卷曲为主,有少量的α-螺旋和β-片层,少见β-转角;推测OmpA蛋白有5个B细胞优势抗原表位区域、2个糖基化位点。本研究为进一步分析禽波氏杆菌免疫机理、制备单克隆抗体和设计表位疫苗等奠定理论基础。 相似文献
8.
猪圆环病毒3型(PCV3)是一种在世界范围内分布较广泛的新型猪病毒,其发病机制尚不清楚。为系统分析PCV3 Capsid(Cap)蛋白序列和结构特点,收集Gen Bank登录的来自不同国家和地区的210条PCV3全基因组和Cap蛋白基因序列,筛选获得46株PCV3代表序列;采用生物信息学(Bio Edit和MAGE 6.0等)技术分析PCV3 Cap序列同源性、结构特点、系统进化和热点突变位点情况。结果表明:46株PCV3 Cap序列同源性为97.66%～100%,PCV3 Cap预测的8个B细胞和10个T细胞表位中均存在序列突变,其中B细胞抗原表位E (109～146位氨基酸)和T细胞抗原表位D(72～81位氨基酸)存在的突变位点数最多。研究发现26株属于PCV3a,20株属于PCV3b。PCV3与PCV2 Cap序列比较分析发现,PCV3 Cap在NLS区域内有6处突变(R11K、R12K、H26M、R27K、V31A和R34K); PCV3 Cap序列间比对分析发现,PCV3b Cap序列出现突变的几率(17/20,85%)高于PCV3a (19/26,73%)。而从临床表现有繁殖障碍和死胎的样品中检测出的PCV3序列均存在S77T和I150L突变位点;从伴有发热、肺炎的仔猪以及肺匀浆样本检测出的PCV3序列中均有R10K突变位点,这些特定的突变位点与临床症状的关系需后续深入研究。本研究结果为PCV3基因功能、进化、预防和控制等研究提供重要序列信息。 相似文献
9.
10.
《中国预防兽医学报》2016,(7)
为了解四川地区腹泻仔猪群中猪圆环病毒Ⅱ型(PCV2)的感染情况,本实验采用PCR方法对病料样品进行PCV2检测,并对分离的病毒DNA进行测序及生物信息学分析。结果显示PCV2感染率为71.6%(312/436),阳性结果中肠系膜淋巴结检出率为100%(312/312)。将扩增并测序的19条PCV2序列与Gene Bank中23株参考序列比较分析,构建了系统进化树,结果表明四川省腹泻仔猪群感染PCV2基因型主要为PCV2b、PCV2d基因型。针对PCV2高频突变ORF2基因编码的Cap蛋白氨基酸序列进行生物信息学分析,结果显示主要抗原表位氨基酸位点存在替换,进而引起抗原漂移,这可能促使PCV2免疫原性发生改变。本研究为四川地区PCV2的防制奠定了基础。 相似文献
11.
B W Ritchie F D Niagro K S Latimer W L Steffens D Pesti R P Campagnoli P D Lukert 《American journal of veterinary research》1992,53(9):1512-1518
Adult umbrella cockatoos, Moluccan cockatoos, African grey parrots, and a yellow-headed Amazon parrot were inoculated IM or SC with beta-propiolactone-treated psittacine beak and feather disease (PBFD) virus. Thirty- to 45-day-old African grey parrot, umbrella cockatoo, and sulphur-crested cockatoo chicks also were vaccinated with the same inoculum. The hemagglutination inhibition (HI) and agar-gel diffusion tests were used to assay for post-vaccination development of anti-PBFD virus antibodies. All adult vaccinates seroconverted and had increases in HI and precipitating antibodies. The vaccinated chicks had increased concentrations of HI antibodies, but precipitating antibodies could not be detected. To demonstrate that chicks from vaccinated hens are protected from PBFD virus challenge, 3 African grey parrot chicks and 2 umbrella cockatoo chicks from vaccinated hens and 1 African grey parrot chick and 1 umbrella cockatoo chick from nonvaccinated hens were exposed to purified PBFD virus. Chicks from the vaccinated hens remained clinically normal during the 50-day test period. Chicks from the nonvaccinated hens developed clinical and histologic lesions of PBFD. Infected tissues from these birds were confirmed to contain viral antigen, using immunohistochemical staining techniques. The PBFD virus was recovered from the affected birds. These findings indicate that adult and 30- to 45-day-old psittacine birds will seroconvert following vaccination with beta-propiolactone-treated PBFD virus. Also, hens inoculated with beta-propiolactone-treated PBFD virus produce chicks that are, at least temporarily, resistant to virus challenge. 相似文献
12.
Schoemaker NJ Dorrestein GM Latimer KS Lumeij JT Kik MJ van der Hage MH Campagnoli RP 《Avian diseases》2000,44(2):470-478
This paper describes the signs, clinical pathology, and postmortem findings in 14 young African grey parrots (Psittacus erithacus erithacus) that were naturally infected with psittacine beak and feather disease (PBFD) virus (psittacine circovirus). All but two of the parrots had severe leukopenia at clinical presentation. Two other parrots also had severe anemia. All birds died within 3 wk after presentation. Postmortem examination documented liver necrosis in 11 of 14 birds and secondary bacterial or fungal infections in 9 of 14 birds. Tests for Chlamydia psittaci, polyomavirus, and Salmonella sp. were negative. PBFD viral infection could be demonstrated in all birds by polymerase chain reaction. Supporting evidence of PBFD viral infection was gathered by histologic examination of the bursa of Fabricius, electron microscopy, and DNA in situ hybridization. Electron microscopic examination of both the bursa of Fabricius and liver revealed virus particles resembling circovirus. DNA in situ hybridization of six liver tissue samples confirmed the presence of PBFD virus and excluded the presence of avian polyomavirus. Our findings suggest that a specific presentation of peracute PBFD viral infection, characterized by severe leukopenia, anemia, or pancytopenia and liver necrosis in the absence of feather and beak abnormalities, may occur in young African grey parrots. 相似文献
13.
Psittacine beak and feather disease (PBFD) is a common viral disease of wild and captive psittacine birds characterized by symmetric feather loss and beak deformities. The causative agent, beak and feather disease virus (BFDV), is a small, circular single-stranded DNA virus that belongs to the genus Circovirus. BFDV can be detected by PCR or the use of haemagglutination (HA) and haemagglutination inhibition (HI) assays that detect antigen and antibodies respectively. Erythrocytes from a limited number of psittacine species of Australian origin can be used in these tests. In South Africa, the high cost of these birds makes them difficult to obtain for experimental purposes. Investigation into the use of erythrocytes from African Grey parrots and Brown-headed parrots yielded positive results showing the haemagglutinating activity of their erythrocytes with purified BFDV obtained from confirmed clinical cases of the disease. The HA activity was further confirmed by the demonstration of HI using BFDV antiserum from three different African Grey parrots previously exposed to the virus and not showing clinical signs of the disease. 相似文献
14.
B W Ritchie F D Niagro K S Latimer W L Steffens D Pesti J Ancona P D Lukert 《American journal of veterinary research》1991,52(11):1804-1809
Psittacine beak and feather disease (PBFD) virus was recovered from the feces and crop washings from various species of psittacine birds diagnosed with PBFD. High concentrations of the virus also could be demonstrated in feather dust collection from a room where 22 birds with active cases of PBFD were being housed. The virions recovered from the feces, crop, and feather dust were confirmed to be PBFD virus by ultrastructural, physical, or antigenic characteristics. Virus recovered from the feather dust and feces hemagglutinated cockatoo erythrocytes. The specificity of the agglutination was confirmed by hemagglutination inhibition, using rabbit antibodies against PBFD virus. During the test period, 26% (8 of 31) of the birds screened were found to be excreting PBFD virus in their feces, and 21% (3 of 14) of crop washings were positive for PBFD virus. Some birds in the sample group had active cases of diarrhea, whereas others had normal-appearing feces. Diarrhea was found to be the only significant indicator of whether a bird was likely to be excreting virus from the digestive tract. These findings suggest that exposure of susceptible birds to PBFD virus may occur from contact with contaminated feather dust, feces, or crop secretions. Viral particles that were morphologically similar to parvovirus (20- to 24 nm-icosahedral nonenveloped virions) also were recovered from feces of some of the birds. 相似文献
15.
Rahaus M Wolff MH 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2003,50(8):368-371
Psittacine beak and feather disease (PBFD) is the most common viral disease of wild and captive psittacine birds. Here, we designed the first survey to investigate the existence of subclinical infections and the distribution of the causative agent named beak and feather disease virus (BFDV) inside the population of captive psittacine birds in Germany. DNA was isolated from feathers of 146 symptom-free birds from 19 different genera (all psittaformes) taken from 32 independent breeders from all over Germany. The presence of BFDV was analysed by performing polymerase chain reaction assays. Fifty-eight (39.2%) samples were found to be positive for BFDV. As expected, there was no significant predominance of one sex to be infected with BFDV. 相似文献
16.
K S Latimer P M Rakich I M Kircher B W Ritchie F D Niagro W L Steffens P D Lukert 《Journal of veterinary diagnostic investigation》1990,2(3):204-207
Thirty-five birds that died with naturally acquired psittacine beak and feather disease (PBFD) were necropsied to identify extracutaneous viral inclusions. Inclusions were found in various tissue sections from 34 of 35 birds. By immunoperoxidase staining, intranuclear and intracytoplasmic inclusion bodies were shown to contain PBFD viral antigen. Inclusion-bearing lesions were widely disseminated but often closely associated with the alimentary tract. Lesions within the palate, esophagus, crop, intestine, bursa of Fabricius, and liver probably serve as sources for viral shedding into the feces. 相似文献
17.
Kiatipattanasakul-Banlunara W Tantileartcharoen R Katayama K Suzuki K Lekdumrogsak T Nakayama H Doi K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(6):527-529
Three sulphur-crested cockatoos (Cacatua galerita) were diagnosed as psittacine beak and feather disease (PBFD). Histopathology of the feather pulp and follicles showed intracytoplasmic botryoid clusters or granular inclusion bodies in epithelial cells and macrophages. Electron microscopy revealed multiple cytoplasmic clusters of electron dense viral particles corresponding to the inclusions. PBFD virus (circovirus) DNA-specific product was detected from formalin-fixed paraffin-embedded feathers by nested polymerase chain reaction (PCR) method. 相似文献
18.
SUMMARY Simple and sensitive haemagglutination and haemagglutination Inhibition assays were developed for psittacine beak and feather disease (PBFD) virus and serum antibody, respectively. The assays were used in the examination of samples from 73 birds clinically affected with PBFD. High antigen titres (log2 9 to log2 12) were detected In feathers, faeces and cloacal contents of PBFD-affected birds. Antigen was not detected In either faecal or feather samples from 20 normal galahs (Eolophus roselcapillus) and 9 normal sulphur crested cockatoos (Cacatua galerita). After kaolin treatment and haemadsorption of serum, haemagglutination inhibition (HI) antibody titres could not be detected in serum from 42 PBFD-affected birds, whereas serum HI titres from 64 normal psittacine birds ranged from less than log2 1 to log2 8. Serum and yolk HI antibody responses of 6 PBFD virus-inoculated layer hens were measured. Pre-inoculation chicken sera contained high concentrations of non-specific haemagglutination inhibitors (not detected in chloroform-extracted yolk), which were removed by kaolin treatment and haemadsorption. 相似文献
19.
K S Latimer P M Rakich W L Steffens I M Kircher B W Ritchie F D Niagro P D Lukert 《Veterinary pathology》1991,28(4):300-304
The nature of feather inclusions was characterized in 32 psittacine birds (30 cockatoos, one peach-faced lovebird (Agapornis roseicollis), and one red-lored Amazon parrot (Amazona autumnalis autumnalis] with naturally-acquired psittacine beak and feather disease. Intranuclear inclusions within feather epithelial cells and intracytoplasmic inclusions within macrophages in the feather epithelium and pulp cavity contained psittacine beak and feather disease viral antigen when stained by the avidin-biotin complex immunoperoxidase technique. Ultrastructurally, inclusions were observed primarily within macrophages and to a lesser extent within epithelial cell nuclei. Macrophage inclusions appeared as paracrystalline arrays of viral particles. Intranuclear inclusions were less well defined, although scattered viral particles were present. Intracytoplasmic and intranuclear particles in ultrastructural preparations were identified by colloidal gold labeling as psittacine beak and feather disease virus. Feather epithelium was more frequently and severely involved in the disease process than was adjacent follicular epithelium. Plucked feathers with an intact epidermal collar and feather epithelium were preferred to follicular biopsies for histopathologic examination. 相似文献
20.
Y Sanada N Sanada M Kubo 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1999,61(9):1063-1065
Psittacine beak and feather disease (PBFD) was diagnosed in an umbrella cockatoo (Cacatua alba) with severe feather dystrophy and loss. Electron microscopically, the intranuclear and intracytoplasmic inclusion bodies observed by light microscopy were composed of viral particles forming paracrystalline arrays, whorls, semicircles or concentric circles. Recovered viral particles from the skin and feather follicle tracts were icosahedral and 15 to 20 nm in diameter. 相似文献