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1.
为了克隆从患肺炎犊牛和健康犊牛上分离的多杀性巴氏杆菌血红素结合受体(heme acquisition system receptor,HasR)基因并进行序列分析,分别以致犊牛肺炎和健康犊牛携带的多杀性巴氏杆菌DNA为模板,通过PCR反应扩增出血红素结合受体基因的全部序列,PCR产物纯化后克隆到pMD19-T载体上,经菌液PCR和酶切鉴定后进行序列测定。各菌株HasR基因序列比对结果发现,Pm-BS-a、Pm142-x-4、Pm142-x-3、Pm149-x、Pm-BS-d的序列之间亲源关系较近,而Pm149-xby与参考菌株Pm70的同源性较高。健康牛源的多杀性巴氏杆菌与致犊牛肺炎多杀性巴氏杆菌之间HasR基因的同源性非常高,说明致犊牛肺炎多杀性巴氏杆菌是一种条件性致病菌,其引起的犊牛肺炎可能属内源性感染。 相似文献
2.
为确定疑似禽霍乱病例病原种类及其病原基因型,本研究采用细菌分离技术对病原菌进行实验室分离培养,应用传统方法和分子生物学方法对分离细菌进行鉴定,并应用PCR扩增和基因序列分析对分离细菌进行基因分型。结果显示,分离菌具有多杀性巴氏杆菌（Pasteurella multocida,Pm）典型培养特征,菌落形态和菌体染色特征、生理生化特性均与多杀性巴氏杆菌相符;PCR扩增到457 bp的基因片段。采用5对分型引物对分离菌进行基因分型显示,仅有A型引物扩增到大小1 050 bp的目的基因片段,序列分析也显示分离菌荚膜基因与A型多杀性巴氏杆菌参考菌株荚膜特异性基因同源性高达97.6%~100.0%,系统进化与A型多杀性巴氏杆菌处于同一进化分支。结果表明,疑似禽霍乱病例病原为A型多杀性巴氏杆菌,本研究结果将为禽霍乱的防控提供参考资料。 相似文献
3.
青海省互助某羊场藏系绵羊发生肺炎疾病,为了快速准确诊断藏羊肺炎发病的致病病原微生物,及时防控和治疗,采集病死绵羊肺样品,利用PCR分子生物学方法进行鉴定与生物信息学分析。经过分子鉴定,此羊场引发肺病的病原是绵羊肺炎支原体(Mycoplasma ovipneumoniae, MO);测序结果显示样品中鉴定的为同一株病原,鉴定的菌株的16S rRNA基因与参考序列EU265779的同源性达到99.86%。同时遗传进化树显示,鉴定的菌株与GenBank中的绵羊肺炎支原体聚成一大支,其关系最近,在进化角度证实此次鉴定的确实为绵羊肺炎支原体。结果表明,此羊场引发肺炎疾病的病原是绵羊肺炎支原体,提示要对羊群进行相关的病原学研究和流行病学调查,为针对性地对细菌性病原进行对症治疗提供参考。 相似文献
4.
本研究旨在对江苏某牧场引发犊牛呼吸道综合征的多杀性巴氏杆菌进行分离鉴定，并分析其遗传进化关系及耐药情况。首先对引起犊牛呼吸道症状致犊牛死亡案例进行剖检，发现肺脏肿大，肺门淋巴结肿胀、出血，肺部形成多处结节，然后取肺门淋巴结进行细菌分离培养与鉴定，并进行16S rRNA扩增、测序分析、血清型鉴定和耐药性分析。质谱鉴定和测序分析结果显示，该菌株与A型多杀性巴氏杆菌属于同一亚群，其序列与已经发布的CQ2、CQ7菌株同源性较近。药敏结果显示，该菌株对林可霉素耐药，对其余13种常用抗生素具有高度敏感性。以上结果表明，引发该牧场犊牛肺炎的病原为A型多杀性巴氏杆菌，且与国内报道的A型多杀性巴氏杆菌同源性较高，提示A型多杀性巴氏杆菌在国内牧场具有较广泛的流行性。本研究为犊牛呼吸道综合征的病原学调查和多杀性巴氏杆菌流行病学提供数据参考。 相似文献
5.
为鉴定临床疑似鸭多杀性巴氏杆菌感染肉鸭的病原菌，本试验通过细菌分离培养、菌体形态观察、细菌生化鉴定、16S rRNA基因测序分析、细菌种特异性鉴定、荚膜分型鉴定和动物回归试验进行鉴定，并通过药敏试验和耐药基因检测进行耐药性分析。结果显示，从患病鸭肝脏组织分离到的细菌在鲜血琼脂培养基中呈现表面光滑凸起、灰白色菌落，为革兰氏阴性短小杆菌，瑞氏染色呈两极浓染；生化鉴定结果显示，分离菌能发酵葡萄糖、蔗糖和甘露醇，硫化氢、氧化酶和吲哚等试验阳性；16S rRNA基因序列系统进化树分析显示，该分离菌与多杀性巴氏杆菌聚为一支，同源性 > 99%；细菌种特异性鉴定结果与多杀性巴氏杆菌相符；荚膜分型鉴定结果仅扩增到约为1 050 bp的目的基因片段，与荚膜血清A型相符；动物回归试验显示，该分离菌有较强的致病性；药敏试验结果显示，该分离菌对羧苄西林、氨苄西林、复方新诺明和四环素等12种药物耐药；经耐药基因PCR检测显示，该分离菌携带Sul1、Sul3、tet（X）和Intl1 4种耐药基因，与药敏表型相符。本试验成功分离到1株鸭源荚膜血清A型多杀性巴氏杆菌，为鸭多杀性巴氏杆菌病的防治提供参考依据。 相似文献
6.
2018年9月广西某羊场部分山羊发生流涕、咳嗽、呼吸困难和体温升高等临床症状的疾病,为确诊发病原因并提供治疗方案,采用病原分离培养、PCR扩增鉴定的方法进行诊断,并对分离菌进行生化鉴定、致病性试验和药敏试验。结果显示,病料在血平板有圆形的小菌落生长,革兰阴性小球短杆菌,而在PPLO培养基上不生长;病料的PCR扩增结果显示绵羊肺炎支原体和多杀性巴氏杆菌均为阳性;所分离到的病原菌经生化鉴定,该菌符合多杀性巴氏杆菌的特性;用多杀性巴氏杆菌种属和D型多杀性巴氏杆菌特异性引物扩增为阳性;致病性试验显示该菌对小鼠有很强的致病性;药敏试验显示该菌对头孢他啶、头孢噻肟、氧氟沙星高度敏感,对复方新诺明、强力霉素、红霉素、青霉素为耐药。结果表明该病是由绵羊肺炎支原体和D型多杀性巴氏杆菌混合感染引起。 相似文献
7.
牛源荚膜血清A型多杀性巴氏杆菌的分离鉴定 总被引：9，自引：4，他引：5
从采集于黑龙江、天津的病牛肺组织中分离到2株病原菌,经菌落形态学观察、培养特性、生化反应和小鼠毒力试验,初步鉴定为多杀性巴氏杆菌,分别命名为Pm-HLJ和Pm-TJ.参考多杀性巴氏杆菌种特异性基因kmt1和荚膜血清型特异性基因hyaD-hyaC、bcbD、dcbF、ecbJ和fcbD,合成引物,通过多重PCR扩增2株菌的种特异性基因和荚膜血清型特异性基因.选取Pm-HLJ的目标PCR产物进行克隆、序列测定、Blast搜索同源序列并且比较分析.结果显示,Pm-HLJ的kmt1基因片段全长460 bp,与GenBank中各血清型kmt1基因同源性均在96.6%以上;荚膜血清型A菌株特异性基因同源性为99.9%;而与其他荚膜血清型B、D、E、F的型特异性基因的同源性均低于50%.由此确认,分离的2株多杀性巴氏杆菌均为荚膜血清A型,这是我国A型多杀性巴氏杆菌引发牛出血性败血症的首例报道. 相似文献
8.
犊牛肺炎多杀性巴氏杆菌的分离与鉴定 总被引：3，自引：0，他引：3
新疆石河子6个规模化奶牛场相继出现出生1～9周龄的犊牛发生体温升高、呼吸困难以肺部感染为主,部分犊牛发生腹泻的疾病,造成90余头犊牛因肺炎死亡.从其中2个发病牛场死亡犊牛采集痛料,经涂片镜检、细菌分离培养、生化鉴定及动物试验,初步鉴定为多杀性巴氏杆菌,分别命名为Pm142-x-3、Pm142-x-4、Pm149-xby、Pm149-x,参考多杀性巴氏杆菌种特异性基因kmtl和荚膜血清型特异性基因hyaD-hyaC、bcbD、dcbF、ecbJ和fcbD序列,合成引物,进行PCR扩增.选取Pm149-x-3的目标PCR产物进行序列测定并分析比较,结果表明,Pm149-x-3的kmtl基因片段全长为460 bp,与GenBank中各血清型kmtl基因核苷酸同源性为99.4%;荚膜血清型A菌株特异性基因核苷酸同源性为97%;而扩增其他荚膜血清型B、D、E、F的型特异性基因均未获得目的条带.由此确认,分离的4株多杀性巴氏杆菌均为荚膜血清A型.药敏试验表明分离的多杀性巴氏杆菌对氧氟沙星、庆大霉素敏感. 相似文献
9.
为了确定石河子地区规模化羊场出现呼吸道症状死亡羊的细菌性病原,采用常规细菌分离鉴定方法、细菌16SrRNA序列分析以及多杀性巴氏杆菌特异性基因kmt,从病变肺组织分离鉴定细菌,利用5个荚膜血清型特异性基因确定其血清型,扩增分离株的16个毒力相关基因,分析分离菌致病性和对常用抗菌药物的耐药性。结果表明,从病羊的病变肺组织中分离鉴定到一株血清D型多杀性巴氏杆菌,具有较强的致病性;携带8个毒力相关基因,其片段序列与NCBI上己公布的多杀性巴氏杆菌参考株同源性在99%以上;分离株对青霉素、林可霉素、庆大霉素、复方新诺明耐药,对其他23种药物敏感。 相似文献
10.
为查明2021年四川省甘孜州某藏绵羊养殖场呼吸道疾病的发病病因,本研究采用PCR方法对送检的5份深部鼻腔棉拭子进行了“山羊副流感病毒3型、牛病毒性腹泻-黏膜病病毒、牛冠状病毒、多杀性巴氏杆菌、溶血性曼氏杆菌、绵羊肺炎支原体、丝状支原体簇成员”等常见呼吸道病原的检测。结果显示：多杀性巴氏杆菌和溶血性曼氏杆菌为阳性,其他病原为阴性;通过细菌分离培养成功获得5株溶血性曼氏杆菌和5株多杀性巴氏杆菌;分型结果显示5株溶血性曼氏杆菌为A2型,5株多杀性巴氏杆菌为D型。结合临床症状,确诊该场藏绵羊的呼吸道疾病是由A2型溶血性曼氏杆菌和D型多杀性巴氏杆菌混合感染所致。 相似文献
11.
On the aetiopathogenesis of Mycoplasma pneumonia in calf 总被引：1，自引：0，他引：1
A pure culture of Mycoplasma (M.) bovis was isolated from calves with respiratory disease, exhibiting the picture of lymphohistiocytic proliferative pneumonia with presence of eosinophil plasmatic cells. A mixed infection of M. bovis and Pasteurella (P.) multocida was demonstrated in calves with exudative pneumonia. Both M. bovis and Haemophilus (H.) somnus were recovered from calves with necrotic pneumonia. All 3 organisms--M. bovis, P. multocida, and H. somnus--were present in cases of exudative-necrotic pneumonia. It was also shown that M. bovis played a primary role in the aetiopathogenesis of respiratory diseases caused by mixed infections. 相似文献
12.
Opsonic monoclonal antibodies against lipopolysaccharide (LPS) antigens of Pasteurella multocida and the role of LPS in immunity. 总被引：5，自引：0，他引：5
B RamdaniAdler
A panel of six monoclonal antibodies (MAbs) produced from mice immunized with Pasteurella multocida (M1404) (Heddleston serotype 2) reacted with homologous lipopolysaccharide, as indicated by enzyme immunoassay and immunoblotting. All six MAbs reacted with serotypes 2 and 5 of the 16 Heddleston serotypes. The reactive epitopes were localized on the bacterial cell surface by immunogold labelling. The antibodies could agglutinate P. multocida only if cells were first treated with 1 N HCl. All six MAbs opsonized P. multocida for phagocytosis by mouse macrophages but were not bactericidal in the presence of complement. They afforded only partial protection against infection in mice. The results, together with those of active immunization experiments with LPS, suggest a subordinate role for LPS in protection from experimental infection in mice. 相似文献
13.
The aim of the study was to examine the ability of Haemophilus somnus and Mycoplasma dispar to induce pneumonia in healthy calves under conditions closely resembling the supposed natural way of infection, viz. by inhalation of aerosol droplets containing the microorganisms. The infections were investigated by recording clinical data, cytokine expression of peripheral blood cells and pathology. Twelve calves were included in the study: Three animals were exposed to H. somnus only, and two to M. dispar only, whereas five were challenged to M. dispar followed by exposure to H. somnus 11-14 days later. Also, one calf was exposed to M. dispar followed by exposure to a sterile saline solution 11 days later, and one calf was only exposed to a sterile saline solution. Just one animal, only challenged with H. somnus, developed a focal necrotizing pneumonia, from which H. somnus was isolated. Thus, the ability of H. somnus and M. dispar to act as primary pathogens under these conditions were minimal and inconsistent.However, a transient rise in body temperature, a marked granulocytosis and increased levels of interleukin-8 in peripheral blood after inoculation with H. somnus indicated a clear systemic response, probably as a consequence of the natural non-specific local and systemic defence mechanisms acting in healthy calves. 相似文献
14.
Palzer A Ritzmann M Majzoub M Wolf G Hermanns W Heinritzi K 《Berliner und Münchener tier?rztliche Wochenschrift》2007,120(11-12):483-489
The aim of this study was to investigate the incidence and influence of different causative organisms involved in the development of pneumonia and bronchopneumonia in pigs. Bronchoalveolar lavage fluid (BALF) from 339 pigs was examined molecular-biologically and culturally. The evaluation considered the following pathogens: Mycoplasma (M.) hyopneumoniae, Mycoplasma (M.) hyorhinis, PRRSV (US-Type), PRRSV (EU-Type), PCV-2, Influenzavirus Type A, alpha-haemolytic Streptococci, beta-haemolytic Streptococci, Pasteurella (P.) multocida, Bordetella (B.) bronchiseptica, Haemophilus (H.) parasuis and Actinobacillus (A.) pleuropneumoniae. This was followed by a pathological-anatomical examination in 168 pigs. Pathological-anatomical examination was evaluated for possible interstitial pneumonia, catarrhal-purulent bronchopneumonia and pleuritis. alpha-haemolytic Streptococci, PCV-2, H. parasuis, M. hyorhinis, M. hyopneumoniae, B. bronchiseptica, PRRSV (US-Type), P. multocida, PRRSV (EU-Type) and Influenzavirus Type A were evident in descending incidence in the BALF. A. pleuropneumoniae were only isolated culturally in two cases in the BALF. The frequency and semiquantitative detection rate in the bacteriological culture of alpha-haemolytic Steptococci, M. hyopneumoniae, P. multocida and B. bronchiseptica correlated significantly with the ascertained clinical evaluation score and the pathological-anatomical examination score. M. hyorhinis and Influenzavirus Type A only correlated with the severity degree of clinical appearance, while PCV-2 and PRRSV (US-Type) correlated with the frequency of pathological-anatomical changes. The higher the clinical score, the higher was the number of animals ascertained with five or more different pathogens. The more different causal agents were isolated in the BALF of one animal, the higher was the average pathological-anatomical examination score. For the diagnosis of pneumonia, especially when analysing facultative pathogens or secondary pathogens, a useful interpretation of analysis results is only possible in connection with a clinical and pathological evaluation. 相似文献
15.
The aim of this study was to evaluate the associations between different pathogens in the development of pneumonia and bronchopneumonia in pigs. Samples of bronchoalveolar lavage fluid from 100 pigs showing no clinical signs and 239 pigs with clinical signs of respiratory disease were examined for Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, US-type porcine respiratory and reproductive syndrome virus (PRRSV), EU-type PRRSV, porcine circovirus type 2 (pcv-2), influenza virus type A, alpha-haemolytic Streptococcus species, beta-haemolytic Streptococcus species, Pasteurella multocida, Bordetella bronchiseptica, Haemophilus parasuis and Actinobacillus pleuropneumoniae. These potential pathogens were detected more frequently in the pigs with respiratory problems than in the pigs with no clinical signs. pcv-2 and alpha-haemolytic streptococci were the pathogens most frequently detected; A pleuropneumoniae was isolated in only two cases. There were more often associations between the organisms in the pigs with clinical signs than in the healthy pigs. In particular, alpha-haemolytic streptococci and M hyopneumoniae were both associated with the presence of M hyorhinis, EU-type PRRSV, P multocida and B bronchiseptica, and alpha-haemolytic streptococci also occurred more often in pigs that were already infected with other pathogens. P multocida and B bronchiseptica were both significantly associated with M hyopneumoniae, alpha-haemolytic streptococci, EU-type PRRSV and US-type PRRSV. 相似文献
16.
Tegtmeier C Uttenthal A Friis NF Jensen NE Jensen HE 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1999,46(10):693-700
During 1 year, the association between microbiological and pathological findings in 72 lungs from calves submitted to the Danish Veterinary Laboratory for diagnostic purposes was studied. All cases were evaluated pathologically and bacteriologically, whereas only 68 cases were examined for the presence of bovine respiratory syncytial virus (BRSV), parainfluenza-3 virus (PI-3 virus) and bovine coronavirus, 62 cases for bovine viral diarrhoea virus (BVD), 45 cases for bovine adenovirus and 51 cases for mycoplasmas. Based on histopathological examination, the cases were diagnosed as fibrinous and/or necrotizing bronchopneumonia, suppurative bronchopneumonia, embolic pneumonia and others. The diagnoses were based on the dominating and most severe lesions in each lung. Haemophilus somnus, Pasteurella multocida, Actinomyces pyogenes, P. haemolytica and BRSV were the most commonly found bacterial and viral lung pathogens, respectively. Pasteurella spp. and H. somnus were often associated with the more severe fibrinonecrotizing type of bronchopneumonia, whereas BRSV was primarily detected in cases of suppurative bronchopneumonia. Mycoplasma bovis was isolated from one case only, whereas M. dispar, M. bovirhinis and Ureaplasma diversum were present, often concomitantly, in the majority of cases. Aspergillus fumigatus was isolated from one case. 相似文献
17.
[目的]牛支原体肺炎是严重危害国内外肉牛养殖业的一种重要疾病,病原混合感染将加剧病情,增加临床治疗难度。本研究通过阐明我国牛支原体肺炎混合感染情况,为探讨更加有效的防控手段提供参考依据。[方法]近4年来采用门诊和出诊的方式,收集了全国范围内35个临床初诊为牛支原体肺炎的牛场病牛样本,进行牛支原体及混合感染细菌的分离和分型鉴定。[结果]确定了这类疾病的细菌学感染特征,表现为牛支原体感染占绝对优势,混合感染模式主要以牛支原体合并多杀性巴氏杆菌A型感染、牛支原体合并和化脓隐秘杆菌感染为主。[结论]经实验室检测证实临床初诊病例绝大部分为牛支原体肺炎,但混合感染很普遍。 相似文献
18.
Lungs from 191 slaughter pigs with gross lesions indicative of enzootic pneumonia of pigs (EPP) and 80 grossly normal lungs, all originating from 9 different herds, were subjected to microbiological and pathological examinations. The microbiological studies included both bacterial and mycoplasmal culture and also testing for Mycoplasma hyopneumoniae antigen in tissue by indirect immunofluorescent technique. M. hyopneumoniae, Pasteurella multocida and Mycoplasma hyorhinis were detected in 83%, 43% and 37% of the pneumonic lungs, respectively. Mycoplasma flocculare was the most frequently isolated organism in the non-pneumonic lungs. The greatest amounts of macroscopic pneumonia (25.2%) were recorded in lungs with all the three agents M. hyopneumoniae, P. multocida and M. hyorhinis present. The amounts of pneumonia in lungs with M. hyopneumoniae alone and in concurrence with P. multocida, were 9.3% and 15.6%, respectively. M. hyorhinis was also, in this study, associated with higher frequency of diffuse pleuritis. These findings indicate that M. hyorhinis might be involved in the pathogenesis of pneumonia in slaughter pigs. Ninety-six per cent of the isolates of P. multocida from pneumonic lungs could be characterized as type A. In the herds which had the most severe pneumonia problems, toxin production was detected in 83% of the P. multocida strains while only 28% were toxigenic in herds with subclinical to moderate pneumonia problems. 相似文献
19.
Pósa R Donkó T Bogner P Kovács M Repa I Magyar T 《Canadian journal of veterinary research》2011,75(3):176-182
The interaction of Bordetella bronchiseptica, toxigenic Pasteurella multocida serotype D, and the mycotoxin fumonisin B(1) (FB(1)) was studied. On day 0 of the experiment, 28 artificially reared 3-day-old piglets were divided into 4 groups (n = 7 each): a control group (A), a group fed FB(1) toxin (B), a group infected with the 2 pathogens (C), and a group infected with the 2 pathogens and fed FB(1) toxin (D). The B. bronchiseptica infection [with 10(6) colony-forming units (CFU)/mL] was performed on day 4 and the P. multocida infection (with 10(8) CFU/mL) on day 16. From day 16 a Fusarium verticillioides fungal culture (dietary FB(1) toxin content 10 mg/kg) was mixed into the feed of groups B and D. In groups C and D, clinical signs including mild serous nasal discharge, sneezing, panting, and hoarseness appeared from day 4, and then from day 16 some piglets had coughing and dyspnea as well. Computed tomography (CT) performed on day 16 demonstrated lung lesions attributable to colonization by B. bronchiseptica in the infected groups. By day 25 the number of piglets exhibiting lesions had increased, and the lesions appeared as well-circumscribed, focal changes characterized by a strong density increase in the affected areas of the lungs. The gross pathological findings confirmed the results obtained by CT. These results indicate that, when combined with dual infection by B. bronchiseptica and P. multocida, dietary exposure of pigs to FB(1) toxin raises the risk of pneumonia and increases the extent and severity of the pathological changes. 相似文献