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首页> 外文期刊>Veterinary Microbiology >Ultrastructural and fluorochromatic changes of Anaplasma marginale exposed to oxytetracycline, imidocarb and enrofloxacin in short-term erythrocyte cultures
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Ultrastructural and fluorochromatic changes of Anaplasma marginale exposed to oxytetracycline, imidocarb and enrofloxacin in short-term erythrocyte cultures

机译：短期红细胞培养中暴露于土霉素，亚氨基脲和恩诺沙星的无性无浆体的超微结构和荧光变化

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Anaplasma marginale causes mild to severe hemoparasitic disease resulting in significant morbidity and mortality in cattle worldwide. In the absence of universally efficacious vaccines, antimicrobial therapy combined with biocontainment and biosecurity strategies are critical to control anaplasmosis. Herein, we compared the effect of oxytetracycline, imidocarb and enrofloxacin on A. marginale isolates in short-term erythrocyte cultures. Electron micrographs detailing antimicrobial-induced changes in rickettsial morphology were scored (0-4) based on ultrastructural changes. These were compared to fluorochromatic changes detected by flow cytometry (FACS) using conversion of hydroethidine (HE) to ethidium bromide (EB) by living organisms to assess viability. A. marginale infectivity in selected cultures was confirmed by subinoculation into susceptible calves. Morphology scores were analyzed using Chi-squared tests and compared to FACS data by ANOVA with isolate, drug and concentration as co-variates in the model. Only the Virginia and Oklahoma isolates exposed to 1.0mug/ml imidocarb and the Oklahoma isolate exposed to 4.0mug/ml enrofloxacin were sterilized following antimicrobial exposure. Rickettsia with morphology scores of 0 had significantly more EB positive cells than inclusions with morphology scores of 4 (p=0.039). There was also a significant association between ultrastructural changes and infectivity (p=0.0047). Furthermore, the percent EB positive cells in the antimicrobial exposed cultures was highly predictive of the probability of infectivity (p=0.0026). This is the first study describing ultrastructural changes in A. marginale following exposure to enrofloxacin and imidocarb. These findings demonstrate that FACS and electron microscopy are useful tools to screen new antimicrobials for the use in anaplasmosis chemotherapy.
机译：边缘无浆膜引起轻度至重度的血液寄生虫病，导致全世界牛的发病率和死亡率显着升高。在缺乏普遍有效的疫苗的情况下，抗微生物治疗与生物控制和生物安全策略相结合对于控制肛门菌病至关重要。在这里，我们比较了土霉素，氨基咪唑和恩诺沙星对短期红细胞培养中的边缘农杆菌的影响。根据超微结构的变化，详细记录了抗菌素引起的立克次体形态变化的电子显微照片（0-4）。将这些与通过流式细胞术（FACS）检测到的荧光色变化进行比较，使用活体将氢乙啶（HE）转化为溴乙锭（EB）来评估生存力。通过将亚接种到易感牛犊中，可以确认所选培养物中的边缘甲虫感染性。使用卡方检验分析形态学得分，并通过ANOVA与分离物，药物和浓度作为模型中的协变量的方差分析与FACS数据进行比较。暴露于1.0mug / ml咪唑威克的弗吉尼亚州和俄克拉荷马州分离株以及暴露于4.0mug / ml恩诺沙星的俄克拉荷马州分离株在暴露于抗菌剂后进行灭菌。形态分数为0的立克次体具有比形态分数为4的包涵体明显更多的EB阳性细胞（p = 0.039）。超微结构变化和感染性之间也存在显着关联（p = 0.0047）。此外，在暴露于抗菌剂的培养物中，EB阳性细胞百分率可高度预测感染的可能性（p = 0.0026）。这是第一个描述暴露于恩诺沙星和咪唑威克后边缘农杆菌超微结构变化的研究。这些发现表明，FACS和电子显微镜是筛选用于厌氧菌化学疗法的新抗菌药物的有用工具。

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