Comparison of Ultrastructure, Extracellular Matrix, and Drug Susceptibility in M. avium subs. hominissuis Biofilms.
Comparison of Ultrastructure, Extracellular Matrix, and Drug Susceptibility in M. avium subs. hominissuis Biofilms.
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DOI：
10.3390/pathogens12121427
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发表时间：
2023-12-08
期刊：
影响因子：
--
通讯作者：
Schorey JS
中科院分区：
文献类型：
--
作者：
McManus WR;Schorey JS
鸟分枝杆菌肺部感染发生在易感个体暴露于环境中的该细菌之后，这种细菌在环境中常常存在于生物被膜中。已经使用了许多方法来生成鸟分枝杆菌的生物被膜，并且不清楚不同的方法是否会产生相似的结构和细胞表型。为了对体外生物被膜超微结构、细胞外基质（ECM）组成以及生物被膜内细菌的药物敏感性进行平行比较，我们使用了两种已发表的方法来生成鸟分枝杆菌生物被膜：在M63培养基中培养四周或用二硫苏糖醇（DTT）处理24小时。扫描电子显微镜显示这两种方法生成的生物被膜超微结构存在差异，包括细胞外基质物质的外观和内部细胞形态的变化，而光学显微镜和用钙荧光白染色表明两种生物被膜都含有纤维素特有的多糖。对生物被膜被酶降解的敏感性进行测量表明，在结构上重要的细胞外基质分子存在差异，DTT生物被膜具有重要的蛋白质成分，在较小程度上还有纤维素成分，而M63生物被膜具有适量的蛋白质、纤维素和DNA成分。两种生物被膜都对阿米卡星和克拉霉素的杀菌作用具有抗性，在几乎所有浓度下，内部细胞的被杀灭速率都比浮游细胞低10倍以上。这些比较表明鸟分枝杆菌在不同条件下生物被膜反应存在差异，但也提示了生物被膜形成的共同特征，包括纤维素产生和抗微生物抗性。
Pulmonary infections with Mycobacterium avium occur in susceptible individuals following exposure to the bacterium in the environment, where it often persists in biofilms. Many methods have been used to generate biofilms of M. avium, and it is unknown whether different approaches generate similar structures and cell phenotypes. To make a parallel comparison of in vitro biofilm ultrastructure, extracellular matrix (ECM) composition, and the drug susceptibility of biofilm resident bacteria, we used two published methods to generate M. avium biofilms: four-week incubation in M63 medium or 24 h exposure to dithiothreitol (DTT). Scanning electron microscopy revealed differences in the biofilm ultrastructure between the two methods, including variation in the appearance of ECM materials and morphology of resident cells, while light microscopy and staining with calcofluor white indicated that both biofilms contained polysaccharides characteristic of cellulose. Measuring the susceptibility of biofilms to degradation by enzymes suggested differences in structurally important ECM molecules, with DTT biofilms having important protein and, to a lesser extent, cellulose components, and M63 biofilms having moderate protein, cellulose, and DNA components. Both biofilms conferred resistance to the bactericidal effects of amikacin and clarithromycin, with resident cells being killed at greater than 10-fold lower rates than planktonic cells at almost all concentrations. These comparisons indicate differences in biofilm responses by M. avium under differing conditions, but also suggest common features of biofilm formation, including cellulose production and antimicrobial resistance.
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影响因子：
3
作者：
Carter, G;Wu, M;Bermudez, LE
通讯作者：
Bermudez, LE
影响因子：
11.8
作者：
Mangione, E J;Huitt, G;Lenaway, D;Beebe, J;Bailey, A;Figoski, M;Rau, M P;Albrecht, K D;Yakrus, M A
通讯作者：
Yakrus, M A
影响因子：
2.2
作者：
Nishiuchi Y
通讯作者：
Nishiuchi Y
影响因子：
4.8
作者：
Ojha, Anil K.;Trivelli, Xavier;Hatfull, Graham F.
通讯作者：
Hatfull, Graham F.
影响因子：
--
作者：
Alhede, Morten;Qvortrup, Klaus;Bjarnsholt, Thomas
通讯作者：
Bjarnsholt, Thomas