We have developed a new type of surface enhanced Raman scattering (SERS) substrate with thiolated graphene oxide (tGO) nanosheets sandwiched between two layers of closely packed plasmonic nanoparticles. The trilayered substrate is built up through alternative loading of interfacially assembled plasmonic nanoparticle arrays and tGO nanosheets, followed by coating the nanoparticle surfaces with poly(ethylene glycol) (PEG). Here tGO plays multifunctional roles as a 2D scaffold to immobilized interfacially assembled plasmonic nanoparticles, a nanospacer to create SERS-active nanogaps between two layers of nanoparticle arrays, and a molecule harvester to enrich molecules of interest via π–π interaction. In particular, the molecule harvesting capability of the tGO nanospacer and the stealth properties of PEG coating on the plasmonic nanoparticles collectively lead to preferential positioning of selective targets such as aromatic molecules and single-stranded DNA at the SERS-active nanogap hotspots. We have demonstrated that an SERS assay based on the PEGylated trilayered substrate, in combination with magnetic separation, allows for sensitive, multiplexed “signal-off” detection of DNA sequences of bacterial pathogens.

Two toluene-degrading strains, T103 and T104, were isolated from rock surface biomass in a freshwater stream contaminated with toluene. The strains exhibit different capacities for degradation of toluene and other aromatic compounds and have characteristics of the genus Mycobacterium. Both are aerobic, rod-shaped, gram-positive, nonmotile, and acid-alcohol fast and produce yellow pigments. They have mainly straight-chain saturated and monounsaturated fatty acids with 10 to 20 carbon atoms and large amounts of tuberculostearic acid that are typical of mycobacteria. Fatty acid analyses indicate that T103 and T104 are different mycobacterial strains that are related at the subspecies level. Their identical 16S rDNA sequences are most similar to Mycobacterium aurum and Mycobacterium komossense, and they constitute a new species of fast-growing mycobacteria. Ecological studies reveal that toluene contamination has enriched for toluene-degrading bacteria in the epilithic microbial community. Strains T103 and T104 play only a small role in toluene degradation in the stream, although they are present in the habitat and can degrade toluene. Other microorganisms are consequently implicated in the biodegradation.