Nancharaiah Yarlagadda V, Joshi Hiren M, Hausner Martina, Venugopalan Vayalam P
Biofouling and Biofilm Processes Section, Water and Steam Chemistry Division, BARC Facilities, Kalpakkam 603 102, Tamil Nadu, India.
Chemosphere. 2008 Mar;71(1):30-5. doi: 10.1016/j.chemosphere.2007.10.062.
This paper describes results of a successful bioaugmentation experiment on aerobic granular sludge using Pseudomonas putida KT2442 cells bearing the TOL (pWWO) plasmid. The methodology was designed to monitor incorporation of the added donor cells into pre-existent microbial granules and the subsequent plasmid transfer to the autochthonous microbial community using shake flask microcosms. Expression of reporter proteins (GFP and DsRed) allowed in situ monitoring of donor cell attachment and plasmid transfer to the recipient cells using confocal laser scanning microscopy. Concomitant with donor integration and transconjugant proliferation in the granules, a significant increase in degradation of benzyl alcohol (used as sole substrate) was observed in the augmented microcosms. In contrast, control microcosms (with non-augmented granules) did not show any noticeable increase in the degradation of the substrate. This study shows that bioaugmentation of aerobic granular sludge via donor colonization and plasmid transfer is feasible for enhanced biodegradation.
本文描述了一项成功的生物强化实验结果，该实验使用携带TOL（pWWO）质粒的恶臭假单胞菌KT2442细胞对好氧颗粒污泥进行处理。该方法旨在通过摇瓶微宇宙监测添加的供体细胞融入预先存在的微生物颗粒的情况，以及随后质粒向本地微生物群落的转移。报告蛋白（绿色荧光蛋白和红色荧光蛋白）的表达使得利用共聚焦激光扫描显微镜能够原位监测供体细胞的附着情况以及质粒向受体细胞的转移。随着供体在颗粒中的整合和转接合子的增殖，在强化的微宇宙中观察到作为唯一底物的苯甲醇的降解显著增加。相比之下，对照微宇宙（含有未强化的颗粒）在底物降解方面没有显示出任何明显的增加。这项研究表明，通过供体定殖和质粒转移对好氧颗粒污泥进行生物强化以增强生物降解是可行的。
