Nanocomposite dermal drug delivery systems based on cellulose nanofibers with grafted titania nanoparticles loaded by two antibiotic medicines from different classes, i.e. tetracycline (TC) and phosphomycin (Phos), were successfully produced by a ''green chemistry'' approach in aqueous media. The influence of a different surface binding mechanism between the drug molecule and modified cellulose nanofibers on the release of the drug and, as a result, on antimicrobial properties against common pathogens Gram-positive, Staphylococcus aureus and Gram-negative Escherichia coli was investigated. The disk diffusion method and broth culture tests using varying concentrations of drugs loaded to nanocomposites were carried out to investigate the antibacterial effects. The influence of UV irradiation on the stability of the obtained nanocomposites and their antibacterial properties after irradiation were also investigated, showing enhanced stability especially for the TC loaded materials. These findings suggest that the obtained nanocomposites are promising materials for the development of potentially useful antimicrobial patches.
Microbial degradation of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (diuron) and mineralization of 4-chloro-2-methylphenoxyacetic acid (MCPA) were studied in soil samples taken from the ballast layers of three Swedish railway embankments. The degradation of diuron followed first-order kinetics and half-lives ranged between 122-365 days. The halflives correlated strongly with microbial biomass estimated by substrate-induced respiration (SIR; R = -0.85; p<0.05) and with the amount of organic matter measured as loss on ignition (R = -0.87; p<0.05). Accumulation of the metabolites DCPMU and DCPU was observed in all samples and these were only detectably degraded in the sample with the highest SIR. Addition of ground lucerne straw to the ballast samples stimulated microbial activity and led to increased formation of metabolites, but further transformation of DCPMU and DCPU was not enhanced. Mineralization of MCPA followed growth-linked kinetics and the time for 50% mineralization was 44.5 ± 7.1 days in samples of previously untreated ballast. In samples of ballast that had been previously treated with the herbicide formulation MCPA 750, the time for 50% mineralization was reduced to 13.7 ±11.3 days. The number of MCPA degraders, quantified using an MPN technique, was clearly increased but highly variable. An average yield of 0.18 cells pg -1 of MCPA was estimated from the kinetic data. The yield estimates correlated with the amount of nitrogen in the ballast, indicating that mineralization of MCPA was nitrogen-limited in the railway embankments studied. This has practical implications for weed control using herbicides on railways.
The use of pesticides on sandy soils and on many non-agricultural areas entails a potentially high risk of water contamination. This study examined leaching of the herbicide 4-chloro-2-methylphenoxyacetic acid (MCPA) after bioaugmentation in sand with differently formulated and stored Sphingobium sp. T51 and at different soil moisture contents. Dry formulations of Sphingobium sp. T51 were achieved by either freeze drying or fluidised bed drying, with high initial cell viability of 67-85 %. Storage stability of T51 cells was related to formulation excipient/carrier and storage conditions. Bacterial viability in the fluidised bed-dried formulations stored at 25 °C under non-vacuum conditions was poor, with losses of at least 97 % within a month. The freeze-dried formulations could be stored substantially longer, with cell survival rates of 50 %, after 6 months of storage at the same temperature under partial vacuum. Formulated and long-term stored Sphingobium cells maintained their MCPA degradation efficacy and reduced MCPA leaching as efficiently as freshly cultivated cells, by at least 73 % when equal amounts of viable cells were used. The importance of soil moisture for practical field bioaugmentation techniques is discussed.
Pesticide residues and their transformation products are frequently found in groundwater and surface waters. This study examined whether adding pesticide-degrading microorganisms simultaneously with the pesticide at application could significantly reduce diffuse contamination from pesticide use. Degradation of the phenoxyacetic acid herbicides MCPA (4-chloro-2-methylphenoxyacetic acid) and 2,4-D (2,4-dichlorophenoxyacetic acid) was studied in soil microcosm experiments after simultaneous spraying of herbicide and herbicide-degrading bacteria on an agricultural soil and on a sand with low degradation potential. The latter represented pesticide use on non-agricultural soils poor in microbial activity. Degradation and possible loss of herbicidal effect were also tested in a system with plants and the amounts of bacteria needed to give satisfactory MCPA-degradation rate and the survival of degrading bacteria in formulated MCPA were determined. The results showed [80-99% degradation of 2,4-D and MCPA in soil within 1 day and [99% within 3 days after inoculation with 10 5 -10 7 herbicide-degrading bacteria g -1 dry weight of soil. Enhanced degradation of MCPA was also obtained in the presence of winter wheat and white mustard without loss of the intended herbicidal effect on white mustard. The survival of an isolated MCPA-degrading Sphingomonas sp. in three realistic concentrations of formulated MCPA was very poor, showing that in practical applications direct contact between the microorganisms and the pesticide formulation must be precluded. The applicability and economic feasibility of the method and the information needed to obtain a useable product for field use are discussed.
In this study we have compared the ability of the organic polymers Ficoll and hydroxyethylcellulose (HEC) and the disaccharides sucrose and trehalose to support cell survival during freeze-drying and subsequent storage of a gram-negative Sphingobium sp. In addition to determination of viability rates, cell integrity was evaluated using lipid peroxidation and RNA quality assays for the different storage conditions and formulation compositions. All formulations resulted in high initial cell survival rates after freeze-drying. However, the disaccharide formulations were superior to the polymer-based formulations in supporting cell survival during storage with the exception of Ficoll that upon storage under vacuum yielded bacterial survival rates equal to that of sucrose. Storage in the presence of both oxygen and moisture was detrimental for bacterial survival in all formulations tested, however, lipid peroxidation or RNA damages were not the controlling mechanisms for cell death in this system. The ability of Ficoll and HEC to support cell survival during freeze-drying show that organic polymers, expected to lack the water replacing capability of e.g. disaccharides, can successfully be used as lyoprotectants. For storage under vacuum conditions we suggest that the intracellular amount of sugars (i.e. trehalose), or other protective native cell components, is sufficient for a basic protection inside the bacteria cell and that the amorphous state is the most important aspect of the formulation excipient. However, when exposed to oxygen and moisture during storage this protection is not sufficient to prevent cell degeneration.
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