Comparison of sampling sites and detection methods for Haemophilus parasuis

Objective To improve the isolation rate and identification procedures for Haemophilus parasuis from pig tissues. Design Thirteen sampling sites and up to three methods were used to confirm the presence of H. parasuis in pigs after experimental challenge. Procedure Colostrum‐deprived, naturally farrowed pigs were challenged intratracheally with H parasuis serovar 12 or 4. Samples taken during necropsy were either inoculated onto culture plates, processed directly for PCR or enriched prior to being processed for PCR. The recovery of H parasuis from different sampling sites and using different sampling methods was compared for each serovar. Results H parasuis was recovered from several sample sites for all serovar 12 challenged pigs, while the trachea was the only positive site for all pigs following serovar 4 challenge. The method of solid medium culture of swabs, and confirmation of the identity of cultured bacteria by PCR, resulted in 38% and 14% more positive results on a site basis for serovars 12 and 4, retrospectively, than direct PCR on the swabs. This difference was significant in the serovar 12 challenge. Conclusion Conventional culture proved to be more effective in detecting H parasuis than direct PCR or PCR on enrichment broths. For subacute (serovar 4) infections, the most successful sites for culture or direct PCR were pleural fluid, peritoneal fibrin and fluid, lung and pericardial fluid. For acute (serovar 12) infections, the best sites were lung, heart blood, affected joints and brain. The methodologies and key sampling sites identified in this study will enable improved isolation of H parasuis and aid the diagnosis of Glässer's disease.     

抗氰烯菌酯假禾谷镰孢突变体的诱导及其生物学特性

为评估引起小麦茎基腐病的病原菌假禾谷镰孢Fusarium pseudograminearum对氰烯菌酯的抗性风险,对5株敏感菌株进行了室内药剂驯化,获得33株抗性突变体,突变频率为16.5%,其对氰烯菌酯的抗性水平范围为7.39～1 665.76倍,3株表现低抗,4株表现中抗,26株表现高抗;发现在myosin-5基因上存在11种抗性突变类型,其中217位的丝氨酸突变为亮氨酸(S217L)、420位的谷氨酸突变为赖氨酸(E420K)和135位的丙氨酸突变为苏氨酸(A135T)为主要突变类型,其比例分别为45.5%、15.2%和9.1%。S217L型抗性突变体的产孢量显著下降,菌丝生长速率和致病力与亲本菌株无显著差异。E420K型抗性突变体的菌丝生长速率和致病力显著下降,产孢量与亲本菌株无显著差异。A135T型抗性突变体的菌丝生长速率和产孢量与亲本菌株无显著差异。研究结果表明假禾谷镰孢在药剂选择压力下易形成氰烯菌酯的抗性群体,对氰烯菌酯存在中到高等的潜在抗性风险,其myosin-5的点突变与其对氰烯菌酯的抗性相关。     

贵州魔芋软腐病菌多基因分子鉴定及其致病力分析

为明确贵州魔芋软腐病菌种类?致病力及分布特点, 采用组织分离法对贵州主要魔芋种植区软腐病样进行病原菌分离, 对icdA, mdh, mtlD, proA, rpoS 等5个管家基因进行了扩增?序列测定, 分别用单基因和多基因联合系统发育树对病菌进行鉴定, 同时采用组织块接种方法测定了不同菌株的致病力?通过组织分离法共分离魔芋软腐病菌株47株; 采用5个管家基因进行分子鉴定, 将病菌分别鉴定为海芋果胶杆菌Pectobacterium aroidearum?胡萝卜果胶杆菌Pectobacterium carotovorum和方中达迪基氏菌Dickeya fangzhongdai 3个种, 其中海芋果胶杆菌P.aroidearum为贵州魔芋软腐病主要致病菌, 占分离菌株的70%, 广泛分布在多个地区; 其次为方中达迪基氏菌D. fangzhongdai, 占分离菌株的28%, 也普遍存在于贵州各魔芋种植区; 胡萝卜果胶杆菌P. carotovorum最少, 占分离菌株的2%?致病力测定结果表明, 菌株间致病力存在一定的差异, 其中海芋果胶杆菌不同菌株之间致病力差异较大, 低?中?高致病力菌株都有, 方中达迪基氏菌差异较小, 仅有中?高致病力菌株?本研究确定了贵州魔芋软腐病菌种类?致病力及在贵州的分布特点, 首次报道了海芋果胶杆菌?方中达迪基氏菌是贵州魔芋软腐病的主要病原菌, 进一步加深了对魔芋软腐病及其发生流行的认识, 为软腐病的科学防控提供了科学依据?     

我国小麦茎基腐病菌致病力比较分析

为明确我国不同种、地理来源和毒素化学型小麦茎基腐病菌的致病力分化情况,采用纸塔法对来自全国9个省市80个采样点分离的224株小麦茎基腐病菌进行致病力分析。结果表明,不同种镰刀菌的致病力不同,黄色镰刀菌Fusarium culmorum,禾谷镰刀菌F.graminearum,假禾谷镰刀菌F.pseudograminearum及亚洲镰刀菌F.asiaticum致病力强于其他种。F.culmorum致病力显著高于F.pseudograminearum和F.asiaticum,而F.pseudograminearum,F.graminearum及F.asiaticum三者之间无显著性差异。中华镰刀菌F.sinensis,木贼镰刀菌F.equiseti,锐顶镰刀菌F.acuminatum致病力较弱,三者间苗期致病力无显著性差异;多数省份F.pseudograminearum群体间致病力无显著差异,仅山东F.pseudograminearum群体的致病性显著低于河南群体;此外,产毒类型为3ADON的F.pseudograminearum群体致病力显著高于15ADON群体。     

苹果轮纹病菌对吡唑醚菌酯的敏感性及Cytb基因序列分析

为了明确苹果轮纹病菌Botryosphaeria dothidea对吡唑醚菌酯的敏感性和吡唑醚菌酯的靶标基因序列, 采用菌丝生长速率法测定了水杨肟酸对B. dothidea 菌丝生长的作用, 探讨了添加或不添加水杨肟酸的情况下病原菌对吡唑醚菌酯敏感性的变化; 并测定了不同产区的80株B. dothidea 对吡唑醚菌酯的敏感性以及440株B. dothidea 对吡唑醚菌酯的抗性; 然后, 扩增并分析了具有不同敏感性的菌株的细胞色素b基因 (Cytb) 序列?结果表明:不同浓度水杨肟酸对菌丝生长的抑制作用不同?添加40 μg/mL水杨肟酸不影响吡唑醚菌酯的EC50?菌株的敏感性频率符合近似正态分布, 各产区菌株对吡唑醚菌酯的敏感性没有显著差异, 吡唑醚菌酯平均EC50为(2.95±2.11) μg/mL, 没有检测到抗性菌株?靶标基因序列分析表明, Cytb基因在F129?G137和G143位密码子上没有产生点突变, 首次发现在143位密码子后有内含子插入?     

Susceptibility pattern of bacterial isolates in equine ulcerative keratitis: Implications for empirical treatment at a university teaching hospital in Sydney

Corneal ulceration is a common ophthalmic condition in horses. It is frequently caused by trauma to the corneal surface, followed by secondary infection by commensal or pathogenic organisms including Streptococcus equi subspecies zooepidemicus, Pseudomonas aeruginosa and Staphylococcus spp. Emerging antimicrobial resistance amongst these organisms has raised the need for appropriate antimicrobial therapy selection, to optimise treatment efficacy while minimising further antimicrobial resistance. Medical records of 38 horses presented at the University Veterinary Teaching Hospital Camden for ulcerative keratitis between 2010 and 2020 were reviewed to identify those with positive bacterial cultures and antimicrobial susceptibility profiles (13/38). Common susceptibility patterns were identified and used to guide the empirical treatment of equine bacterial corneal ulcers. Pseudomonas spp. (64.3%), Streptococcus equi subspecies zooepidemicus (14.3%) and Actinobacillus spp. (14.3%) were most commonly identified. Susceptibility to amikacin, gentamicin and ciprofloxacin was observed in 100%, 66.7% and 85.7% Pseudomonas spp. isolates respectively. Resistance to polymyxin B and neomycin occurred in 85.7% and 71.4% of Pseudomonas spp., respectively. All Streptococcus equi subspecies zooepidemicus organisms in this study were susceptible to ceftiofur, cephalexin, penicillin and ampicillin, while they were all resistant to gentamicin, neomycin, enrofloxacin and marbofloxacin. Predominating in this study, Pseudomonas spp. maintained overall aminoglycoside susceptibility despite some emerging resistance, and good fluoroquinolone susceptibility. High resistance to Polymyxin B could have arisen from its common use as first-line therapy for bacterial corneal ulcers. Although further research is required, these new findings about predominant bacteria in equine corneal ulceration in the Camden region and their antimicrobial susceptibility patterns can be used to guide the empirical treatment of bacterial corneal ulcers in horses.