Metabolomics Australia, Bio21 Molecular Science and Biotechnology Institute, Faculty of Science, University of Melbourne, 3052, Parkville, VIC, Australia; Land and Water, CSIRO, PO Box 56, Highett, VIC 3190, Australia.
Land and Water, CSIRO, PO Box 56, Highett, VIC 3190, Australia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Sep 1;966:179-86. doi: 10.1016/j.jchromb.2014.02.058. Epub 2014 Mar 12.
Water supply biofilms have the potential to harbour waterborne diseases, accelerate corrosion, and contribute to the formation of tuberculation in metallic pipes. One particular species of bacteria known to be found in the water supply networks is Pseudomonas sp., with the presence of Pseudomonas putida being isolated to iron pipe tubercles. Current methods for detecting and analysis pipe biofilms are time consuming and expensive. The application of metabolomics techniques could provide an alternative method for assessing biofilm risk more efficiently based on bacterial activity. As such, this paper investigates the application of metabolomic techniques and provides a proof-of-concept application using liquid chromatography coupled with time-of-flight mass spectrometry (LC-ToF-MS) to three biologically independent P. putida samples, across five different growth conditions exposed to solid and soluble iron (Fe). Analysis of the samples in +ESI and -ESI mode yielded 887 and 1789 metabolite features, respectively. Chemometric analysis of the +ESI and -ESI data identified 34 and 39 significant metabolite features, respectively, where features were considered significant if the fold change was greater than 2 and obtained a p-value less than 0.05. Metabolite features were subsequently identified according to the Metabolomics Standard Initiative (MSI) Chemical Analysis Workgroup using analytical standards and standard online LC-MS databases. Possible markers for P. putida growth, with and without being exposed to solid and soluble Fe, were identified from a diverse range of different chemical classes of metabolites including nucleobases, nucleosides, dipeptides, tripeptides, amino acids, fatty acids, sugars, and phospholipids.
供水生物膜有可能藏匿水源性疾病、加速腐蚀，并促成金属管道中结核的形成。一种已知存在于供水管网中的特殊细菌是假单胞菌属，假单胞菌属的绿脓假单胞菌被分离到铁管结核中。目前检测和分析管道生物膜的方法既耗时又昂贵。代谢组学技术的应用可以提供一种替代方法，根据细菌活性更有效地评估生物膜风险。因此，本文研究了代谢组学技术的应用，并使用液相色谱-飞行时间质谱联用仪（LC-ToF-MS）对三个生物学独立的绿脓假单胞菌样本在五个不同的生长条件下暴露于固铁和可溶铁（Fe）的情况进行了概念验证应用。正、负离子模式下的样品分析分别产生了 887 和 1789 个代谢物特征。正、负离子模式数据的化学计量学分析分别鉴定出 34 和 39 个显著的代谢物特征，如果变化倍数大于 2 且获得的 p 值小于 0.05，则认为特征显著。根据代谢组学标准倡议（MSI）化学分析工作组，使用分析标准品和标准在线 LC-MS 数据库，对代谢物特征进行了鉴定。从核苷、核苷、二肽、三肽、氨基酸、脂肪酸、糖和磷脂等不同化学类别的代谢物中，鉴定出了在有无固铁和可溶铁暴露情况下，绿脓假单胞菌生长的可能标志物。
