猪多杀性巴氏杆菌皮肤坏死毒素亚单位蛋白的制备及免疫原性研究     

何海  郝成武  凌晨  张飞  候凤  贺笋 《中国畜牧兽医》2019,46(11):3396-3403

为获得针对猪萎缩性鼻炎主要病原体——多杀性巴氏杆菌皮肤坏死毒素的高免疫原性抗原，本试验构建、表达并验证该毒素的亚单位蛋白抗原，将猪多杀性巴氏杆菌毒素PMT基因与pMD19-T载体进行连接、转化，经序列分析鉴定后，分别使用BamH Ⅰ、Hind Ⅲ与Blp Ⅰ限制性内切酶将其酶切为3个基因片段。将片段1（Tox1）、片段2（Tox2）和片段3（Tox3）分别亚克隆至原核表达载体pET32-b、pET32-a和pET32-b中，构建3个重组表达载体。将重组表达载体转化E.coli BL21（DE3）感受态细胞，经IPTG诱导后，分别进行SDS-PAGE与Western blotting检测，并使用小鼠与豚鼠初步研究其免疫原性。结果显示，构建的3个基因片段长度分别为776、409与410 bp，与GenBank中相关序列具有高度同源性；3个蛋白片段表达正常，表达量分别达到379.95、447.62与459.82 μg/mL，SDS-PAGE验证条带分别为75、77与53 ku；因3个片段位置不同，仅Tox3有Western blotting检测条带，与理论预测相符；使用3种表达蛋白免疫小鼠与豚鼠后，二免后14 d，血清经试剂盒检测阳性率达到100%，对二免后14 d小鼠攻毒保护率达到93%。本研究成功构建了PMT的3个亚单位活性片段，且具有较好的免疫原性。  相似文献   

鸵鸟嗜肺性巴氏杆菌病的诊治     

张志成  吴结革 《动物医学进展》2003,24(5):116-118

嗜肺性巴氏杆菌在兽医病原学中的地位并不受重视 ,对动物的致病作用也很少见报道 ,在鸵鸟病中更是罕见。文章针对一起鸵鸟慢性死亡的病例 ,经临床症状、病理剖检和实验室检验 ,诊断为嗜肺性巴氏杆菌所致 ,并提出了相应的有效防治方案  相似文献   

Comparative pharmacokinetics of florfenicol in healthy and Pasteurella multocida‐infected Gaoyou ducks     

Weixuan Lan  Xia Xiao  Yongjia Jiang  Lijie Jiang  Xu Zhao  Zhaoyang Yu  Biwen Zhu  Congcong Li  Lin Bian  Zhiqiang Wang 《Journal of veterinary pharmacology and therapeutics》2019,42(3):355-360

Pasteurella multocida is the causative agent of fowl cholera, and florfenicol (FF) has potent antibacterial activity against P. multocida and is widely used in the poultry industry. In this study, we established a P. multocida infection model in ducks and studied the pharmacokinetics of FF in serum and lung tissues after oral administration of 30 mg/kg bodyweight. The maximum concentrations reached (C_max) were lower in infected ducks (13.88 ± 2.70 μg/ml) vs. healthy control animals (17.86 ± 1.57 μg/ml). In contrast, the mean residence time (MRT: 2.35 ± 0.13 vs. 2.27 ± 0.18 hr) and elimination half‐life (T_½β: 1.63 ± 0.08 vs. 1.57 ± 0.12 hr) were similar for healthy and diseased animals, respectively. As a result, the area under the concentration curve for 0–12 hr (AUC_0–12 hr) for FF in healthy ducks was significantly greater than that in infected ducks (49.47 ± 5.31 vs. 34.52 ± 8.29 μg hr/ml). The pharmacokinetic differences of FF in lung tissues between the two groups correlated with the serum pharmacokinetic differences. The C_max and AUC_0–12 hr values of lung tissue in healthy ducks were higher than those in diseased ducks. The concentration of FF in lung tissues was approximately 1.2‐fold higher than that in serum both in infected and healthy ducks indicating that FF is effective in treating respiratory tract infections in ducks.  相似文献   

菪可轮蒲水煎液对多杀性巴氏杆菌体内外抑菌活性研究     

高艳  郝宝成  吴金措姆  格桑卓嘎  班旦  四郎玉珍  邢小勇  梁云  杨志伟  王兰芳  胡永浩  梁剑平  拉巴次旦 《动物医学进展》2019,(7):43-48

研究菪可轮蒲水煎液对多杀性巴氏杆菌的体内外抑菌活性作用。用传统水煎法制备菪可轮蒲水煎液,用HPLC法测定菪可轮蒲水煎液中龙胆苦苷浓度,并用龙胆苦苷浓度标定复方药物浓度,分别采用牛津杯法和宏量肉汤稀释法测定菪可轮蒲水煎液对多杀性巴氏杆菌的抑菌圈直径和最小抑菌浓度(MIC),通过建立死亡率为75%的多杀性巴氏杆菌体内感染小鼠模型,测定浓度为0.0232mg/mL的菪可轮蒲水煎液,在感染前和感染后给药方式下,分别给予0.1、0.3、0.5mL药物时对小鼠多杀性巴氏杆菌感染的预防和治疗作用。结果显示,菪可轮蒲水煎液浓度为0.0464mg/mL时,对多杀性巴氏杆菌的抑菌圈平均直径为13.5mm,MIC值为0.0029mg/mL,经病理剖检、细菌镜检、全自动微生物鉴定及药敏系统(VITEK2COMPACT)细菌鉴定,表明死亡率为75%的多杀性巴氏杆菌体内感染小鼠模型建立成功,且预防组各组和治疗组各组小鼠死亡率依次为58.3%、41.7%、83.3%、66.7%、58.3%和91.7%。综合以上试验结果,菪可轮蒲水煎液对多杀性巴氏杆菌具有良好的体内外抑菌活性,且其预防作用较治疗作用明显。  相似文献   

牛荚膜A型多杀性巴氏杆菌强菌株rpoE蛋白的原核表达及其免疫保护效力研究     

嵇少泽  高云航  勾长龙  张喜庆  王羽  田佳琪  李晨 《中国预防兽医学报》2019,(7):740-743

为研究牛荚膜A型多杀性巴氏杆菌(PM)强菌株rpoE蛋白的免疫保护性,本研究利用PCR方法扩增牛荚膜A型PM的rpoE基因,构建pET-28a-rpoE重组表达质粒,经诱导表达后SDS-PAGE检测结果显示,表达的rpoE蛋白相对分子量约为20 ku,与理论值一致;western blot结果显示该蛋白具有很好的特异性和反应原性;纯化的rpoE蛋白经皮下免疫BALB/c小鼠,间接ELISA检测融合蛋白的免疫原性,结果显示rpoE蛋白可以诱导小鼠产生较高水平的特异性抗体;PM攻毒后,免疫组保护率为70%。本研究获得的牛荚膜A型PM重组rpoE蛋白具有良好的免疫原性,免疫小鼠后能够提供一定的保护,可以作为研发PM亚单位疫苗的候选抗原。  相似文献   

Invasive Pasteurella multocida Infections – Report of Five Cases at a Minnesota Hospital, 2014          下载免费PDF全文

P. Talley  P. Snippes‐Vagnone  K. Smith 《Zoonoses and public health》2016,63(6):431-435

During October 2014, the Minnesota Department of Health was notified of five Hospital A patients with Pasteurella multocida bacteraemia; three had died. Human soft tissue infection with P. multocida typically results from cat or dog bites or scratches. Invasive infection, defined as a P. multocida isolate from a usually sterile site, is rare. We evaluated P. multocida isolations at Hospital A, compared with other Minnesota hospitals to understand invasive infection trends. A case was defined as clinically confirmed P. multocida in a Minnesota resident during 2012–2014. All hospital laboratories were queried; Fisher's exact test was used for comparison. Medical charts were reviewed for 2014 Hospital A patients with P. multocida infections. The Minnesota clinical laboratories survey response rate was 79% (63/80). At Hospital A, proportion of P. multocida isolates from usually sterile sites increased from 0% (0/2) during 2012 to 11% (1/9) during 2013, and to 86% (5/6) during 2014. The proportion of patients with P. multocida isolated from sterile sites was 35% (6/17) at Hospital A compared with 10% (58/583) statewide during 2012–2014 combined (P < 0.05). Among 2014 Hospital A patients with invasive P. multocida infection, all five were men; median age was 70 (range: 44–78) years. Four were temporally clustered within a 33‐day period; three of those had bacteraemia on admission, making hospital acquisition possible in only one. Among five bacteraemia patients, four had cirrhosis and/or skin ulcerations, and three died. The proportion of invasive P. multocida cases was substantially higher at Hospital A during 2014. No epidemiologic links between patients were found. Three had known pet exposure. Collaborative educational efforts of chronically ill pet owners by physicians and veterinarians can acknowledge the health benefits of pet ownership, while minimizing risk for serious invasive zoonotic infections, including those caused by P. multocida.  相似文献   

鸭源多杀性巴氏杆菌对磺胺类药物的耐药性检测及耐药基因分析     

陈红梅  程龙飞  邓辉  刘荣昌  傅光华  施少华  傅秋玲  万春和  郑庆礼  黄瑜  黄小红 《中国畜牧兽医》2019,46(1):223-230

为明确鸭源多杀性巴氏杆菌对磺胺类药物的耐药性及耐药基因的分布特征,试验采用琼脂二倍稀释法对分离自福建、广东和江西等地的89株鸭源多杀性巴氏杆菌进行甲氧苄啶/磺胺甲噁唑的耐受性检测,用PCR法检测各耐药菌株的磺胺类耐药基因(sul1、sul2和sul3),并利用多位点序列分型(MLST)分析耐药菌株间的亲缘关系。结果显示,33株鸭源多杀性巴氏杆菌对甲氧苄啶/磺胺甲噁唑耐受,耐药率为37.1%(33/89);在耐药菌株中,sul1、sul2和sul3基因检出率分别为100%(33/33)、97.0%(32/33)和21.2%(7/33),其中sul1和sul2基因同时检出率为97.0%(32/33),sul1、sul2和sul3基因同时检出率为21.2%(7/33);所有耐药菌株均属于ST129型。以上结果表明,鸭源多杀性巴氏杆菌对磺胺类药物具有一定的耐药性,其中的耐药菌株都属于同一序列型ST129,均携带有sul1耐药基因,且绝大多数耐药菌株同时携带sul1与sul2两种耐药基因,提示在福建、广东和江西等地需慎用磺胺类药物来防治该细菌的感染。  相似文献   

鸭源多杀性巴氏杆菌的分离鉴定及耐药性分析     

陈国权  张旭  阎朝华  周碧君  王开功  程振涛  文明 《中国畜牧兽医》2020,47(2):620-628

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   

致脑膜炎多杀性巴氏杆菌的分离鉴定及全基因组重测序     

王斐  杨洁  吕庆杰  王米雪  刘鹏  张若愚  史聪聪  王雪莹  林琳  华琳  宋文博  梁婉  陈焕春  吴斌  彭忠 《畜牧兽医学报》2022,53(12):4346-4355

旨在了解致脑膜炎多杀性巴氏杆菌的生物学特性及其引起脑部感染的生物学基础。本研究从有神经症状的病死猪脑组织中分离鉴定1株多杀性巴氏杆菌SD001,对其开展血清杀菌试验、胞内存活试验、小鼠感染试验,并利用纳米孔测序技术对其进行全基因组测序。SD001相对于猪其他器官中分离的多杀性巴氏杆菌而言表现出更强的抗血清杀菌的能力,其在小鼠巨噬细胞内存活的能力也较强。小鼠感染试验显示,相同剂量的细菌经血液感染后,SD001在小鼠血液和脑组织中的载菌量显著高于猪肺炎型多杀性巴氏杆菌(P ≤ 0.01),并且SD001造成了小鼠脑组织发生炎症等病理损伤,提示SD001可能造成血流感染并引起小鼠脑膜炎。纳米孔测序显示,SD001的全基因组大小约为2.45 Mb,平均GC含量为40.25%,编码2 281个蛋白以及77个RNA;SD001基因组中含有2条CRISPR结构、5个基因岛、7条前噬菌体序列;毒力基因预测结果显示,SD001含有233个毒力基因;基因分型结果显示,SD001为荚膜：LPS：MLST基因型A:L6:ST10。SD001感染可以引起脑膜炎,其较强的抗血清杀菌能力以及能形成血流感染的能力可能是SD001引起脑部感染的重要原因,在兽医临床关于中枢神经系统感染的诊断中应该纳入多杀性巴氏杆菌感染的可能性。  相似文献   

Protective effect following intranasal exposure of goats to live Pasteurella multocida B:2     

Zamri-Saad M  Ernie ZA  Sabri MY 《Tropical animal health and production》2006,38(7-8):541-546

This study aimed to determine the effect of intranasal exposure to low doses of Pasteurella multocida B:2 on survival of goats challenged with high doses of the same organism. Eighteen goats were selected and divided into three groups. Goats of group 1 were exposed intranasally twice, with a two-week interval, to 7× 10⁶ cfu/ml of live P. multocida B:2. Goats of group 2 were not exposed to P. multocida B:2 but were kept together with the exposed group 1. Goats of group 3 remained as unexposed controls and were kept separated from the other two groups. Serum samples were collected at weekly intervals to determine the antibody levels. At week 5 post exposure, all goats were challenged subcutaneously with 3.7× 10¹⁰ cfu/ml of live P. multocida B:2. Following challenge exposure, 8 (67%) goats (4 goats from each of groups 1 and 2) were killed owing to haemorrhagic septicaemia. Four goats were killed peracutely within 48 h post challenge, while the other four goats were killed acutely between 2 and 4 days post challenge. None of the goats of group 3 were killed for haemorrhagic septicaemia. Goats of groups 1 and 2 showed significantly (p<0.05) higher antibody levels following the first intranasal exposure to P. multocida B:2. However, only group 1 retained the significantly (p<0.05) high antibody levels following a second intranasal exposure, and remained significantly (p<0.05) higher than groups 2 and 3 at the time of challenge. P. multocida B:2 was successfully isolated from various organs of goats that were killed between 1 and 4 days post challenge.  相似文献