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1.
本试验旨在构建一种以不同分支杆菌特异片段为目的基因的多重PCR检测方法。采用GenBank公布的结核分支杆菌RD10序列、牛分支杆菌moaB3序列、鸟分支杆菌16-23SrDNA序列设计并合成特异性引物,扩增产物条带分别为954bp、297bp、119bp。结果显示,三段目的基因都有很高的特异性,对比菌株均无扩增产物出现。对倍比稀释的模板质粒进行检测,该方法的最低检出量为103 copies/μL的DNA模板,该方法特异、敏感、重复性好。首次应用的moaB3基因所得扩增效果良好,成功区分出牛分支杆菌。此多重PCR方法可用于结核分支杆菌、牛分支杆菌、鸟分支杆菌的鉴别和牛结核病的快速临床检测。 相似文献
2.
对豫南地区所采集的76例蛋鸡病料进行鼻气管鸟杆菌分离鉴定和药物敏感性分析。经细菌分离、生化试验,鉴定出2株鼻气管鸟杆菌。利用药敏试验对2株分离株进行了14种常用抗生素的耐药性检测。结果显示:2株鼻气管鸟杆菌均对头孢克肟、丁胺卡那霉素、培氟沙星、环丙沙星、氧氟沙星、强力霉素最敏感;对氟苯尼考、阿莫西林、恩诺沙星、新霉素表现出中度敏感;对链霉素、红霉素、土霉素表现出耐药性。 相似文献
3.
本文报道了从上海动物园棕尾虹雉（Lophophorusimpejanus）体上所采集的鸟虱,经鉴定后,该虫神为头角亚目（Amblycera）短角鸟虱科（Menoponidae）短角鸟虱属（Menopon）一新种,文内对其所具有的分类特征进行了描述,并绘图。 相似文献
4.
本文报道了从上海动物园褐马鸡体上所采到的一种羽虱,根据其特征,经鉴定后,该虫种为头角亚目,短角鸟虱产、短角鸟虱属一新种,文内就其所有的特征进行描述,并绘图。 相似文献
5.
痘病毒感染的物种已经超过230个,这种感染的普遍性给人们提出了一个问题:痘病毒的感染是如何在世界范围内传播的?Gyuranecz等在近期出版的《Journal of Virology》发表了相关论文:利用DNA聚合酶基因,结合4b核心蛋白基因区序列,通过对111个鸟痘病毒自然感染病例的分析获得了痘病毒更加完善的遗传发育框架。这为鸟痘病毒基因提供了一个更加宽泛和完善的遗传分析、测序及分类的方法。 相似文献
6.
1991年在南非的受害肉鸡中分离到了一种多形性格兰氏阴性棒状杆菌 ,当时无法将该菌归入任何一类已知的细菌之中。起初 ,该菌被称为巴氏杆菌样细菌 ,或称为Taxon 2 8,但后来该菌被命名为鼻气管鸟杆菌 (Ornithobacteriumrhinotracheale ,OR)。然而 ,后已证明这并非第一个OR菌株 ;因为 ,据发现 ,分别于 1981年在德国、1983年在比利时以及1987年在匈牙利分离到的菌株都与此南非菌株完全一致。接着就证明了 ,鼻气管鸟杆菌在商品鸡群和野鸟中的存在是全世界的普遍现象。全世界都在蛋中和 1日龄雏鸟中检测到了抗OR母源抗体。一些血清学调查表明… 相似文献
7.
鸡鼻气管炎鸟杆菌属于革兰氏阴性菌,单纯的鼻气管炎鸟杆菌感染往往不引起明显的呼吸道症状,但并发大肠杆菌感染时,可加重鼻气管炎鸟杆菌的感染,并表现为明显的呼吸道症状。2011年12月,河南省信阳市平桥区一养殖场蛋鸡发病,导致大批鸡死亡,造成严重的经济损失,经诊断为鸡鼻气管炎鸟杆菌合并大肠杆菌病,现将诊 相似文献
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为确定云南省易门县某养鸡场大量肉鸡出现流鼻涕、打喷嚏、面部肿胀、食欲下降的原因，采集病死肉鸡鼻腔黏液及肺、气管等组织，处理后提取病原核酸，通过RT-PCR/PCR方法分别检测禽流感病毒（AIV）、传染性喉气管炎病毒（ILTV）、鼻气管鸟杆菌（ORT）、鸡副嗜血杆菌（ICV）。结果显示，仅ORT为核酸阳性，其他病原均为阴性。对检出的ORT阳性样品16S rDNA基因进行序列分析发现，阳性样品与伊朗分离株和土耳其/匈牙利分离株的同源性均在99.7%以上，都处于同一分支内，与国内2014年的报道一致，说明云南省流行的ORT变异较少，仅具有相对短的进化史。本研究对于指导该地区鸡群细菌病防控具有参考意义。 相似文献
9.
鸡鼻气管鸟杆菌病是由鼻气管鸟杆菌引起的鸡和火鸡的一种急性、高度接触性呼吸道传染病。虽然本病在 2 0世纪 90年代才由Ｈｉｍ和ＨａｆｅＴ定性 ,但迄今为止 ,世界许多国家和地区 ,如南美、南非、美国、德国、荷兰、以色列、匈牙利和日本等均已发现本病。鼻气管鸟杆菌不仅能引起鸡和火鸡发生严重的呼吸道疾病 ,而且还造成患病鸡的生产性能明显下降 ,经济损失巨大。目前 ,我国已有病菌分离报道 ,因此 ,应注意本病的防范。1 病原自 1990年以来 ,在许多国家的鸡群中发现了一种当时未知的 ,与鸡和火鸡呼吸道疾病有关的细菌 ,最初入们把这种菌… 相似文献
10.
一种新的禽病—鼻气管鸟杆菌病 总被引：4，自引：0，他引：4
鼻气管鸟杆菌病是近年来发现的一种感染家禽和野乌的呼吸道细菌病，临床上以气囊炎和肺炎为特征，可通过空气水平传播，也可经蛋垂直传播，商品化的鉴定系统及ＰＣＲ可用于该菌的鉴定，谝力至少可分为１２个血清型，适当佐剂制备的灭活苗可减轻该病的临床症状。 相似文献
11.
为探索MAP0862蛋白在分枝杆菌感染鉴别诊断中的作用,研究构建了MAP0862的原核表达map0862-pET32a(+)质粒,并对其进行了诱导表达。以副结核分枝杆菌P10基因组DNA为模板,经PCR方法扩增副结核分枝杆菌map0862基因片段,将其克隆到原核表达载体pET32a(+)中。将重组子转化到大肠杆菌BL21(DE3),在E.coli中诱导表达带组氨酸标签的map0862-pET32a(+)的融合蛋白。结果显示,经IPTG诱导表达出约55 kD的融合蛋白,用副结核阳性血清进行Western blot检测,证明MAP0862重组蛋白具有良好的免疫原性。 相似文献
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13.
Whittington RJ Reddacliff LA Marsh I McAllister S Saunders V 《Australian veterinary journal》2000,78(1):34-37
OBJECTIVES: To determine the frequency of excretion of Mycobacterium avium subsp paratuberculosis in Merino sheep with Johne's disease and to quantify excretion in a group of Merino sheep. DESIGN: A pen and laboratory experiment. PROCEDURE: Seven sheep selected from an affected flock on the basis of acid-fast bacilli in the sheep's faeces were housed and total daily faecal output was collected, weighed and subjected to culture for M avium subsp paratuberculosis. An end-point titration method was used to enumerate viable M avium subsp paratuberculosis in a 15 day pooled sample from five sheep that had acid-fast bacilli in their faeces while housed. RESULTS: Four sheep with subclinical multibacillary Johne's disease excreted M avium subsp paratuberculosis each day for 11 days of cultural observation. A further three sheep were intermittent excreters but lacked other evidence of infection with M avium subsp paratuberculosis. The average number of viable bacteria excreted was 1.09 x 10(8) per gram of faeces while total daily excretion was 8.36 x 10(10) viable M avium subsp paratuberculosis per sheep. Examination of faecal smears stained with Ziehl Neelsen was an unreliable means of assessing daily excretion in individual animals except in those with severe lesions. CONCLUSION: Excretion of M avium subsp paratuberculosis in Merino sheep with multibacillary Johne's disease occurred daily, proving that environmental contamination can be continuous on farms with endemic ovine Johne's disease. Faecal culture is a useful method for detecting infection as it does not appear to be affected by the timing of collection of a sample from sheep with multibacillary disease however, to maximise the sensitivity of disease surveillance using faecal culture, sampling rates should be adjusted to take account of the proportions of multibacillary and paucibacillary cases. 相似文献
14.
牛分枝杆菌(M.bovis)是引起牛结核病的常见病原,而鸟分枝杆菌(M.avium)2型则通常交叉感染牛,但不会导致严重的病变.为鉴定M.bovis和M.avium的免疫交叉反应情况,本研究分别以M.bovis和M.avium 2型的基因组为模板,扩增ag85b基因,分别构建了M.bovis和M.avium的原核和真核表达重组质粒进行表达.以原核表达纯化的两种重组蛋白Ag85b (rAg85b)分别作为包被抗原,交叉检测两种真核重组质粒免疫豚鼠制备的抗血清.结果表明,原核表达的M.bovis和M.avium rAg85b与真核重组质粒免疫豚鼠制备的两种抗血清之间存在较强的免疫交叉反应.研究结果揭示M.bovis和M.avium的Ag85b蛋白存在很强的血清学交叉反应,这将严重干扰M.bovis Ag85b作为候选疫苗的免疫监测. 相似文献
15.
The Mycobacterium avium complex (MAC) encompasses important pathogens in both animals and humans, yet little information is available on the factors required for MAC virulence. An animal isolate, M. avium strain 724 was found to be considerably more virulent in Balb/c mice than a human isolate, M. avium strain A5. To identify the genetic basis of this difference subtractive hybridization was applied, which resulted in the isolation of six DNA fragments unique to strain 724. BLAST searches showed that three sequences belonged to a large gene cluster responsible for biosynthesis of M. avium glycopeptidolipids (GPLs). To reveal the nature of variation between strains in the GPL cluster 27.5kb of a clone containing the A5 serotype-specific GPL (ssGPL) cluster was isolated, sequenced and compared to the corresponding region in other M. avium strains. The ssGPL cluster was highly conserved in the 5' region between all strains and serotypes tested; the 3' region reflects extensive divergence among serotypes including whole gene deletions and insertions of sequences containing open reading frames but lacking identity to any known genes. 相似文献
16.
Basagoudanavar SH Goswami PP Tiwari V Pandey AK Singh N 《Veterinary research communications》2004,28(3):209-224
The full-length open reading frame coding for a potentially immunogenic 35 kDa protein of Mycobacterium avium paratuberculosis was generated using polymerase chain reaction technology. The gene was inserted in-frame into Escherichia coli expression plasmid pQE32. The resulting recombinant plasmid pPMP35 was transformed into E. coli M15. Analysis of the E. coli induced with isopropyl-beta-D-thiogalactopyranoside revealed that the protein accumulated into the cytoplasm as insoluble inclusion bodies. The level of expression of the recombinant 35 kDa protein (P35) was more than 30% of the total protein of E. coli cells. Expression of the recombinant protein was confirmed by immunoblotting. The P35 reacted with a rabbit antiserum raised against a sonicate of M. a. paratuberculosis. The protein was also recognized by serum from a goat with clinical paratuberculosis. Further, a polyclonal antiserum against P35 recognized a 35 kDa band in a membrane fraction of M. a. paratuberculosis. Also, the protein provoked a significant skin reaction in outbred guinea pigs sensitized with M. a. paratuberculosis, as well as in those sensitized with Mycobacterium avium. The results indicate that the 35 kDa protein of M. a. paratuberculosis is a membrane protein, having a role in the cellular immune response. 相似文献
17.
Gioffré A Caimi K Zumárraga MJ Meikle V Morsella C Bigi F Alito A Santángelo MP Paolicchi F Romano MI Cataldi A 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2006,53(1):34-41
A Mycobacterium avium subsp. paratuberculosis expression library in lambda ZAP was screened with immunized mice sera. One clone was selected, sequenced and further characterized. The sequence analysis of the hypothetical open-reading frame (ORF) predicts a protein of 20.8 kDa with a probable signal sequence compatible with Cys-acylation at Cys24, characteristic of lipoproteins. In consequence, the protein was termed Lpp34. Recombinant expression of Lpp34 was achieved by cloning the lpp34 gene into the histidine-tag expression vector pRSET-A. Western blot analysis showed a protein band with a molecular weight of 34 kDa. The native protein was localized in the membrane fraction of M. avium subsp. paratuberculosis and extracted in the detergent phase of Triton X-114. Southern blot and polymerase chain reaction showed that the gene is absent from all the non-M. avium complex mycobacterial genomes tested. Humoral reactivity using bovine sera demonstrated that this protein is widely recognized by both the infected and non-infected animals. This could partly be due to the conserved sequence in close-related environmental bacteria such as M. avium subsp. avium and to the presence of a conserved epitope in other bacteria such as Escherichia coli. In conclusion, these findings show that Lpp34 is a membrane protein and a putative lipoprotein present in M. avium complex mycobacteria and absent in the M. tuberculosis complex. 相似文献
18.
通过PCR扩增分离保存的8株禽波氏杆菌、3株兔支气管败血波氏杆菌及1株猪支气管败血波氏杆菌菌株的23S rRNA基因的片段(710bp),分别克隆到载体pMD18-T后测序。利用BLAST工具进行同源性搜索;用DNAStar分析软件进行同源核苷酸序列的多重比较分析并构建禽波氏杆菌菌株的系统生物进化树。结果显示,8株禽波氏杆菌核苷酸序列之间同源性为99.2%～99.7%,与GenBank中收录的NC_010645株禽波氏杆菌核苷酸序列同源性为92.4%～92.5%,与兔支气管败血波氏杆菌和猪支气管败血波氏杆菌的核苷酸序列同源性均为98.5%～99.2%。国内分离的禽波氏杆菌菌株和支气管败血波氏杆菌菌株均与国外分离菌株在遗传基因上有一定的差异。结果表明,23S rRNA序列分析可以作为鉴定禽波氏杆菌的一种快速简便的方法。 相似文献
19.
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
20.
Schrenzel M Nicolas M Witte C Papendick R Tucker T Keener L Sutherland-Smith M Lamberski N Orndorff D Heckard D Witman P Mace M Rimlinger D Reed S Rideout B 《Veterinary microbiology》2008,126(1-3):122-131
Mycobacterium avium subsp. avium and Mycobacterium intracellulare are primary causes of mycobacteriosis in captive birds throughout the world, but little is known about how they are transmitted. To define the local epidemiology of infection, we strain-typed 70 M. avium subsp. avium and 15 M. intracellulare culture isolates obtained over a 4-year period from captive birds. Typing was performed using randomly amplified polymorphic DNA (RAPD) PCR, amplified fragment length polymorphic (AFLP) fragment analyses, and for a subset of isolates, DNA sequencing of a segment of the 16S-23S rRNA internal transcribed spacer region. Six strain clusters comprising 43 M. avium subsp. avium, isolates were identified; 42 isolates had unique typing patterns, including all M. intracellulare isolates. Phylo-geographical analyses using RAPD and AFLP fingerprints and animal confinement histories showed no correlation between housing of infected birds and mycobacterial strain-type, except for two animals. The diversity of M. avium subsp. avium and M. intracellulare isolates and minimal evidence for bird-to-bird transmission suggest that environmental reservoirs may be important sources of infection in captivity. 相似文献