Browsing by Author "Hattingh, Melanie"
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- ItemCiprofloxacin-eluting nanofibers inhibits biofilm formation by pseudomonas aeruginosa and a Methicillin-resistant Staphylococcus aureus(Public Library of Science, 2015-04) Ahire, Jayesh J.; Neveling, Deon P.; Hattingh, Melanie; Dicks, Leon Milner Theodore, 1961-; MicrobiologyPseudomonas aeruginosa and Staphylococcus aureus are commonly associated with hospital- acquired infections and are known to form biofilms. Ciprofloxacin (CIP), which is normally used to treat these infections, is seldom effective in killing cells in a biofilm. This is mostly due to slow or weak penetration of CIP to the core of biofilms. The problem is accentuated by the release of CIP below MIC (minimal inhibitory concentration) levels following a rapid (burst) release. The aim of this study was to develop a drug carrier that would keep CIP above MIC levels for an extended period. Ciprofloxacin was suspended into poly(D,Llactide) (PDLLA) and poly(ethylene oxide) (PEO), and electrospun into nanofibers (CIP-F). All of the CIP was released from the nanofibers within 2 h, which is typical of a burst release. However, 99% of P. aeruginosa PA01 cells and 91% of S. aureus Xen 30 cells (a methicillinresistant strain) in biofilms were killed when exposed to CIP-F. CIP levels remained above MIC for 5 days, as shown by growth inhibition of the cells in vitro. The nanofibers were smooth in texture with no bead formation, as revealed by scanning electron and atomic force microscopy. A single vibration peak at 1632 cm-1, recorded with Fourier transform infrared spectroscopy, indicated that CIP remained in crystal form when incorporated into PDLLA: PEO. No abnormalities in the histology of MCF-12A breast epithelial cells were observed when exposed to CIP-F. This is the first report of the inhibition of biofilm formation by CIP released from PDLLA: PEO nanofibers.
- ItemCopper-containing anti-biofilm nanofiber scaffolds as a wound dressing material(Public Library of Science, 2016) Ahire, Jayesh J.; Hattingh, Melanie; Neveling, Deon P.; Dicks, Leon Milner Theodore, 1961-Copper particles were incorporated into nanofibers during the electrospinning of poly-D,L-lactide (PDLLA) and poly(ethylene oxide) (PEO). The ability of the nanofibers to prevent Pseudomonas aeruginosa PA01 and Staphylococcus aureus (strain Xen 30) to form biofilms was tested. Nanofibers containing copper particles (Cu-F) were thinner (326 ± 149 nm in diameter), compared to nanofibers without copper (CF; 445 ± 93 nm in diameter). The crystalline structure of the copper particles in Cu-F was confirmed by X-ray diffraction (XRD). Copper crystals were encapsulated, but also attached to the surface of Cu-F, as shown scanning transmission electron microscopy (STEM) and transmission electron microscopy (TEM), respectively. The copper particles had no effect on the thermal degradation and thermal behaviour of Cu-F, as shown by thermogravimetric analysis (TGA) and differential scanning calorimeter (DSC). After 48 h in the presence of Cu-F, biofilm formation by P. aeruginosa PA01 and S. aureus Xen 30 was reduced by 41% and 50%, respectively. Reduction in biofilm formation was ascribed to copper released from the nanofibers. Copper-containing nanofibers may be incorporated into wound dressings.
- ItemThe effect of lactic acid bacteria and fungi on the malting of barley(Stellenbosch : Stellenbosch University, 2013-03) Hattingh, Melanie; Dicks, Leon Milner Theodore; Stellenbosch University. Faculty of Science. Dept. of Microbiology.ENGLISH ABSTRACT: Barley malt is the predominant raw material for beer brewing world-wide. To meet consumer demand, a constant high quality malt product is required. Malt quality is determined by the degree of substrate hydrolysis during germination and mashing which serves as fermentable substrates for alcoholic fermentation during brewing. It is often difficult to sustain malt of high quality due to inconsistent malt batches and poor germination capacities of dormant barley. External additives such as chemicals and gibberellic acid have been used to overcome these difficulties but are unwanted in the beverage industry. Maltsters are consequently always in search of alternative solutions. Microbes produce diverse enzymes which can contribute to substrate hydrolysis during germination. The development of such starter cultures might provide a natural and economically feasible alternative to augment barley germination. Starter culture technology has been employed in the malting industry, although the main focus has been to improve the microbial stability of malt. The exploitation of cultures with hydrolytic capabilities to augment barley germination is consequently largely unexplored. The aim of this study was to develop a starter culture which can contribute to the enzymatic degradation of barley polymers. Geotrichum spp. and Lactobacillus plantarum were isolated from substrates rich in polymers present in barley and screened for enzymatic capabilities. Geotrichum spp. produced cellulase, xylanase, protease and β-glucanase activities, while L. plantarum harboured cell-bound and extracellular α-amylase activities. These cultures were added in different combinations during the malting of Erica and SSG 564 cultivars, but did not enhance germination significantly. Improved malt parameters did not correlate with microbial enzyme activities and the data were not repeatable. Preliminary plate assays could thus not be used to predict enzyme production in a malting environment. Cell-free supernatants with known enzyme activities of Aspergillus sp., Trichoderma reesei and Rhizopus sp. significantly enhanced malt quality. To our knowledge, the use of fungal supernatant to augment malt modification is a novel concept. Supernatant is more convenient than starter cultures and will aid to deliver more constant malt products than live cultures, as known enzyme levels are added.