Seoktae Kang

Antibacterial effects of carbon nanotubes: size does matter!

We provide the first evidence that the size (diameter) of carbon nanotubes (CNTs) is a key factor governing their antibacterial effects and that the likely main CNT-cytotoxicity mechanism is cell membrane damage by direct contact with CNTs. Experiments with well-characterized single-walled carbon nanotubes (SWNTs) and multiwalled carbon nanotubes (MWNTs) demonstrate that SWNTs are much more toxic to bacteria than MWNTs. Gene expression data show that in the presence of both MWNTs and SWNTs, Escherichia coli expresses high levels of stress-related gene products, with the quantity and magnitude of expression being much higher in the presence of SWNTs.

Electronic-Structure-Dependent Bacterial Cytotoxicity of Single-Walled Carbon Nanotubes

Single-walled carbon nanotubes (SWNTs) have been previously observed to be strong antimicrobial agents, and SWNT coatings can significantly reduce biofilm formation. However, the SWNT antimicrobial mechanism is not fully understood. Previous studies on SWNT cytotoxicity have concluded that membrane stress (i.e., direct SWNT-bacteria contact resulting in membrane perturbation and the release of intracellular contents) was the primary cause of cell death. Gene expression studies have indicated oxidative stress may be active, as well. Here, it is demonstrated for the first time how SWNT electronic structure (i.e., metallic versus semiconducting) is a key factor regulating SWNT antimicrobial activity. Experiments were performed with well-characterized SWNTs of similar length and diameter but varying fraction of metallic nanotubes. Loss of Escherichia coli viability was observed to increase with an increasing fraction of metallic SWNTs. Time-dependent cytotoxicity measurements indicated that in all cases the majority of the SWNT antimicrobial action occurs shortly after (<15 min) bacteria-SWNT contact. The SWNT toxicity mechanism was investigated by in vitro SWNT-mediated oxidation of glutathione, a common intracellular thiol that serves as an antioxidant and redox state mediator. The extent of glutathione oxidation was observed to increase with increasing fraction of metallic SWNTs, indicating an elevated role of oxidative stress. Scanning electron microscopy images of E. coli in contact with the SWNTs demonstrated electronic structure-dependent morphological changes consistent with cytotoxicity and glutathione oxidation results. A three-step SWNT antimicrobial mechanism is proposed involving (i) initial SWNT-bacteria contact, (ii) perturbation of the cell membrane, and (iii) electronic structure-dependent bacterial oxidation.

Characteristics and fates of soluble microbial products in ceramic membrane bioreactor at various sludge retention times

The formation and fate of soluble microbial products (SMP) in membrane bioreactor (MBR) was investigated at various sludge retention times (SRT) for 170 days. The SMP concentration was estimated by feeding glucose, which could be completely degraded, and by measuring the dissolved organic carbon (DOC) of the effluent from MBR. Under the conditions of SRT of 20 days, influent DOC of 112 mg/l and HRT of 6 h, the produced SMP was 4.7 mg DOC/l of which 57% was removed or retained by the membrane. DOC of MBR supernatant increased during 100 days and then gradually decreased. Specific UV absorbance showed that the accumulated compounds had a portion of larger, more aromatic, more hydrophobic and double-bond-rich organics, which originated from the decayed biomass. Molecular weight distributions of SMP in MBR supernatant showed that the acclimated microorganisms in a long SRT could decompose high molecular weight organics, it caused the shift of molecular weight distributions of SMP to a lower range. During the operation period, enumeration of active cells in the MBR showed that microbial inhibitions by accumulated SMP was not observed.

Bioinspired single bacterial cell force spectroscopy.

We developed a method for preparing live bacterial cell probes for atomic force microscopy (AFM) using a bioinspired polydopamine wet adhesive. Microscopic examinations with bacterial and yeast cells indicated that cells were successfully glued to the end of the AFM cantilevers and remained viable for the duration of the force measurements. Interaction forces measured with live single-cell microorganism probes differed markedly from those obtained with glutaraldehyde-fixed microorganism probes. Interaction forces between live cell probes and quartz surfaces involved both repulsive steric forces and multimodal weak adhesion forces, which were attributed to the soft exocellular polymeric layers and the heterogeneity of the cell membrane surfaces.

Ultrafiltration Membranes Incorporating Amphiphilic Comb Copolymer Additives Prevent Irreversible Adhesion of Bacteria

We examined the resistance to bacterial adhesion of a novel polyacrylonitrile (PAN) ultrafiltration membrane incorporating the amphiphilic comb copolymer additive, polyacrylonitrile-graft-polyethylene oxide (PAN-g-PEO). The adhesion of bacteria (E. coli K12) and the reversibility of adhered bacteria were tested with the novel membrane, and the behavior was compared to a commercial PAN ultrafiltration membrane. Under static (no flow) bacterial adhesion tests, we observed no bacterial adhesion to the PAN/PAN-g-PEO membrane at all ionic strengths tested, even with the addition of calcium ions. In contrast, significant adhesion of bacterial cells was observed on the commercial PAN membrane, with increased cell adhesion at higher ionic strengths and in the presence of calcium ions. Under crossflow filtration conditions, initial bacterial deposition rate increased with ionic strength and with addition of calcium ions for both membranes, with generally lower bacterial deposition rate with the PAN/PAN-g-PEO membrane. However, deposited bacteria were readily removed (between 97 and 100%) from the surface of the PAN/PAN-g-PEO membrane upon increasing the crossflow and eliminating the permeate flow (i.e., no applied transmembrane pressure), suggesting reversible adhesion of bacteria. In contrast, bacterial adhesion on the commercial PAN membrane was irreversible, with approximately 50% removal of adhered bacteria at moderate ionic strengths (10 and 30 mM) and less than 25% removal at high ionic strength (100 mM). The resistance to bacterial adhesion of the PAN/PAN-g-PEO membrane was further analyzed via measurement of interaction forces with atomic force microscopy (AFM). No adhesion forces were detected between a carboxylated colloid probe and the PAN/PAN-g-PEO membrane, while the probe exhibited strong adhesion to the commercial PAN membrane, consistent with the bacterial adhesion tests. The exceptional resistance of the PAN/PAN-g-PEO membrane to bacterial adhesion is attributable to steric repulsion imparted by the dense brush layer of polyethylene oxide (PEO) chains.

Antimicrobial biomaterials based on carbon nanotubes dispersed in poly(lactic-co-glycolic acid)

Biomaterials that inactivate microbes are needed to eliminate medical device infections. We investigate here the antimicrobial nature of single-walled carbon nanotubes (SWNTs) incorporated within the biomedical polymer poly(lactic-co-glycolic acid) (PLGA). We find Escherichia coli and Staphylococcus epidermidis viability and metabolic activity to be significantly diminished in the presence of SWNT-PLGA, and to correlate with SWNT length and concentration (<2% by weight). Up to 98% of bacteria die within one hour on SWNT-PLGA versus 15-20% on pure PLGA. Shorter SWNTs are more toxic, possibly due to increased density of open tube ends. This study demonstrates the potential usefulness of SWNT-PLGA as an antimicrobial biomaterial.

SWNT-MWNT hybrid filter attains high viral removal and bacterial inactivation.

We describe the concept and demonstrate the efficacy of a novel SWNT-MWNT hybrid filter for the removal and inactivation of microbial pathogens from water. The filter is composed of a thin SWNT layer (0.05 mg cm(-2)) on top of a thicker MWNT layer (0.27 mg cm(-2)) supported by a microporous support membrane. The SWNT-MWNT filter exhibits high log removal of several model viruses (MS2, PRD1, and T4 bacteriophages) by depth filtration, which predominantly takes place in the thicker and more uniform MWNT layer. The filter removes all bacteria by a sieving mechanism, with the top SWNT layer providing high levels of inactivation of model bacteria (Escherichia coli K12 and Staphylococcus epidermidis), as well as microbes from river water and treated wastewater effluent. The dual-layer SWNT-MWNT filter lays the framework for new possibilities in point-of-use water filtration.

Effect of Membrane Surface Properties During the Fast Evaluation of Cell Attachment

Abstract The biofouling potential is one of the important factors to design and to select membranes for water and wastewater treatment. In this investigation, the effect of membrane surface properties during the attachment of S. cerevisiae cells was examined using a laboratory‐scale membrane filtration cell enabling direct microscopic observation of microbial cell deposition. The experimental results from 6 commercially available membranes showed that the initial adhesion rate, k d , was affected by the zeta potentials, hydrophobicity, and roughness of membrane surfaces. The k d value was significantly lower at the membrane which had more negative, hydrophilic, and smooth surfaces. The results will be helpful to minimize the time for selecting membranes in different situations, and for testing the performance of newly designed membranes.

Influence of shear on the production of extracellular polymeric substances in membrane bioreactors

Shear is used to control fouling in membrane bioreactor (MBR) systems. However, shear also influences the physicochemical and biological properties of MBR biomass. The current study examines the relationship between the level of shear and extracellular polymeric substance (EPS) production in MBRs. Two identical MBRs were operated in parallel where the biomass in one reactor was exposed to seven times greater shear forces. The concentrations of floc-associated and soluble EPS were monitored for the duration of the experiment. The stickiness of extracted floc-associated EPS from each reactor was also characterized using atomic force microscopy. A mathematical model of floc-associated and soluble EPS production was applied to quantitatively describe changes in EPS production with shear. Biomass grown in a high shear environment has lower floc-associated EPS production compared to biomass grown in a lower shear environment. This decrease in floc-associated EPS production also corresponds to a decrease in soluble EPS production, which can be explained by both the lower concentration of floc-associated EPS and the production of stickier floc-associated EPS that is more erosion resistant in the high shear reactor. This research suggests that mechanical stresses can have a significant impact on the production rates of floc-associated and soluble EPS-key parameters governing membrane fouling in MBRs.

Impact of conditioning films on the initial adhesion of Burkholderia cepacia

Bacterial initial adhesion to inert surfaces in aquatic environments is mainly governed by the surface properties of the substratum, which can be altered significantly by the formation of conditioning films. Bacteria were tested for ability to adhere to bare glass slides and to slides coated with alginate, bovine serum albumin (BSA), or Suwannee River natural organic matter (SR-NOM). Three Burkholderia cepacia strains with different extracellular polymeric substance (EPS) secretion capacities were tested. The surface roughness of the slides was measured by atomic force microscopy (AFM), but its effect on bacterial initial adhesion was not significant. Our results showed the degree (number of cells per cm(2)) of initial adhesion among the three strains of B. cepacia was not significantly different, indicating that B. cepacia surface EPS did not impact adhesive capacity in the conditions tested. Depending on the conditioning film types and ionic strength conditions, conditioning film coatings can either enhance or reduce bacterial initial adhesion. Bacterial adhesion to bare slides and to alginate or SR-NOM coated slides increased with increasing ionic strength; however, a similar trend was not observed on BSA coated slides. Although BSA coated slides were the most hydrophobic and had the lowest negative surface charge among the surfaces tested, bacterial adhesion was not enhanced by the BSA coating. The extended Derjaguin-Landau-Verwey-Overbeek (DLVO) theory was applied to explain bacterial adhesion to solid surfaces.