Anna Żywicka

Modification of bacterial cellulose through exposure to the rotating magnetic field

The aim of the study was to assess the influence of rotating magnetic field (RMF) on production rate and quality parameters of bacterial cellulose synthetized by Glucanacetobacter xylinus. Bacterial cultures were exposed to RMF (frequency f=50Hz, magnetic induction B=34mT) for 72h at 28°C. The study revealed that cellulose obtained under RMF influence displayed higher water absorption, lower density and less interassociated microfibrils comparing to unexposed control. The application of RMF significantly increased the amount of obtained wet cellulose pellicles but decreased the weight and thickness of dry cellulose. Summarizing, the exposure of cellulose-synthesizing G. xylinus to RMF alters cellulose biogenesis and may offer a new biotechnological tool to control this process. As RMF-modified cellulose displays better absorbing properties comparing to non-modified cellulose, our finding, if developed, may find application in the production of dressings for highly exudative wounds.

Increased water content in bacterial cellulose synthesized under rotating magnetic fields

ABSTRACT The current study describes properties of bacterial cellulose (BC) obtained from Komagataeibacter xylinus cultures exposed to the rotating magnetic field (RMF) of 50 Hz frequency and magnetic induction of 34 mT for controlled time during 6 days of cultivation. The experiments were carried out in the customized RMF exposure system adapted for biological studies. The obtained BC displayed an altered micro-structure, degree of porosity, and water-related parameters in comparison to the non-treated, control BC samples. The observed effects were correlated to the duration and the time of magnetic exposure during K. xylinus cultivation. The most preferred properties in terms of water-related properties were found for BC obtained in the setting, where RMF generator was switched off for the first 72 h of cultivation and switched on for the next 72 h. The described method of BC synthesis may be of special interest for the production of absorbent, antimicrobial-soaked dressings and carrier supports for the immobilization of microorganisms and proteins.

Biochemical and cellular properties of Gluconacetobacter xylinus cultures exposed to different modes of rotating magnetic field

Abstract The aim of the present study was to evaluate the impact of a rotating magnetic field (RMF) on cellular and biochemical properties of Gluconacetobacter xylinus during the process of cellulose synthesis by these bacteria. The application of the RMF during bacterial cellulose (BC) production intensified the biochemical processes in G. xylinus as compared to the RMF-unexposed cultures. Moreover, the RMF had a positive impact on the growth of cellulose-producing bacteria. Furthermore, the application of RMF did not increase the number of mutants unable to produce cellulose. In terms of BC production efficacy, the most favorable properties were found in the setting where RMF generator was switched off for the first 72 h of cultivation and switched on for the further 72 h. The results obtained can be used in subsequent studies concerning the optimization of BC production using different types of magnetic fields including RMF, especially.

Increased yield and selected properties of bacterial cellulose exposed to different modes of a rotating magnetic field

Rotating magnetic field (RMF) is an interesting alternative to conventional bacterial cellulose (BC) production methods. The BC synthesis processes may be affected by RMF, which facilitates the transfer of oxygen and nutrients from the media to the microbial cells. RMF may also directly influence the various physical and chemical properties of BC. The main aim of the present study was to evaluate the impact of the RMF on the BC in regard to its yield and material properties. The correlation between the efficiency of polymer production and the different time of exposure to the RMF was also analyzed to determine the conditions of lower energy consumption during the cellulose formation process. It was found that the Gluconacetobacter xylinus cultures exposed to the RMF for a half of the time of the entire cellulose production process (72 h), considering the results obtained in controls, synthesized BC more effectively than bacteria continuously exposed to the RMF for 144 h. Furthermore, the application of the RMF, regardless of the exposure mode, did not negatively affect the polymer material properties. It was concluded that the use of the RMF may provide a novel technique for altering cellulose biogenesis and may be used in multiple biotechnological applications.

Modification of Bacterial Cellulose with Quaternary Ammonium Compounds Based on Fatty Acids and Amino Acids and the Effect on Antimicrobial Activity

In the present work, bacterial cellulose (BC) membranes have been modified with bioactive compounds based on long chain dimer of C18 linoleic acid, referred to as the dilinoleic acid (DLA) and tyrosine (Tyr), a natural amino acid capable of forming noncovalent cation-π interactions with positively charged ethylene diamine (EDA). This new compound, [EDA][DLA-Tyr], has been synthesized by simple coupling reaction, and its chemical structure was characterized by 1H NMR and Fourier transform infrared spectroscopy. The antimicrobial activity of a new compound against S. aureus and S. epidermidis, two cocci associated with skin and wound infections, was assessed. The [EDA][DLA-Tyr] impregnated BC exhibited strong and long-term antimicrobial activity against both staphylococcal species. The results showed a 57-66% and 56-60% reduction in S. aureus and S. epidermidis viability, respectively, depending on [EDA][DLA-Tyr] concentration used. Importantly, [EDA][DLA-Tyr] molecules were released gradually from the BC pellicle, while a reference antibiotic, erythromycine (ER), did not show any antibacterial activity against S. aureus and S. epidermidis after 48 h of soaking in deionized water. Thus, a combination of [EDA][DLA-Tyr] and BC could be a promising new class of wound dressing displaying both biocompatibility and antimicrobial activity.

Time Dependent Influence of Rotating Magnetic Field on Bacterial Cellulose

The aim of the study was to assess the influence of rotating magnetic field (RMF) on the morphology, physicochemical properties, and the water holding capacity of bacterial cellulose (BC) synthetized by Gluconacetobacter xylinus. The cultures of G. xylinus were exposed to RMF of frequency that equals 50 Hz and magnetic induction 34 mT for 3, 5, and 7 days during cultivation at 28°C in the customized RMF exposure system. It was revealed that BC exposed for 3 days to RMF exhibited the highest water retention capacity as compared to the samples exposed for 5 and 7 days. The observation was confirmed for both the control and RMF exposed BC. It was proved that the BC exposed samples showed up to 26% higher water retention capacity as compared to the control samples. These samples also required the highest temperature to release the water molecules. Such findings agreed with the observation via SEM examination which revealed that the structure of BC synthesized for 7 days was more compacted than the sample exposed to RMF for 3 days. Furthermore, the analysis of 2D correlation of Fourier transform infrared spectra demonstrated the impact of RMF exposure on the dynamics of BC microfibers crystallinity formation.

Bacterial cellulose yield increased over 500% by supplementation of medium with vegetable oil

Bacterial cellulose (BC), produced by Komagataeibacter xylinus, has numerous applications to medicine and industry. A major limitation of BC use is relatively low production rates and high culturing media costs. By supplementing culture media with 1% vegetable oil, we achieved BC yield exceeding 500% over the yield obtained in standard media. BC properties were similar to cellulose cultured in standard methods with regard to cytotoxicity but displayed significantly higher water swelling capacity and mechanical strength. As we demonstrated herein, this significantly increased BC yield is the result of microscopic and macroscopic physiochemical processes reflecting a complex interaction between K. xylinus biophysiology, chemical processes of BC synthesis, and physiochemical forces between BC membranes, oil and culturing vessel walls. Our findings have significant translational implications to biomedical and clinical settings and can be transformative for the cellulose biopolymer industry.

A.D.A.M. test (Antibiofilm Dressing's Activity Measurement) — Simple method for evaluating anti-biofilm activity of drug-saturated dressings against wound pathogens

In the present article, we propose a simple Antibiofilm Dressings Activity Measurement (A.D.A.M.) test that allows to check in vitro a dressings suitability against biofilm-related wound infections. To perform the test, three agar discs are covered with biofilm formed by the tested pathogen after which they are assembled one over another in the form of an agar plug and placed in the well of a 24-well plate. The top disc is covered with the analyzed dressing and the entire set is incubated for 24h. During this time, the investigated antimicrobial substance is released from the dressing and penetrates to subsequent biofilm-covered agar discs. Biofilm reduction is measured using 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) spectrometric assay and the results are compared to untreated control samples (agar plug covered with biofilm and without the dressing/or with a passive dressing placed on the top disc). Furthermore, in order to standardize the differences in penetrability of the drugs released from active dressings the results can be expressed as a dimensionless value referred to as the Penetrability Index. In summary, A.D.A.M. test is simple, cheap, can be performed practically in every clinical laboratory and takes no more time than routine microbiological diagnostics. Apart from measuring the released drugs activity, the A.D.A.M. test allows to assess drug penetrability (across three agar discs), reflecting real wound conditions, where microbes are frequently hidden under the necrotic tissue or cloth. In conclusion, the A.D.A.M. test produces a high volume of data that, when analyzed, can provide a researcher with a valuable hint concerning the applicability of active dressings against specific biofilm pathogens in a particular setting.

Influence of milk, milk fractions and milk proteins on the growth and viability of mastitis-causing Staphylococcus aureus strain

Abstract The aim of this study was to investigate the impact of milk and milk fractions (cell-reduced, skim and whey) obtained from different cows on the growth rate of mastitis-causing Staphylococcus aureus strain at low inoculum density, simulating the early phase of intramammary infection. The association of the selected milk proteins, including α-lactalbumin, β-lactalbumin, lactoferrin, bovine serum albumin, γ-globulin and casein with the bacterial growth was also analysed. Twelve Polish Holstein-Friesian cows having no history of mastitis during the previous and current lactation were selected for this study. The S. aureus strain used in this study was isolated from a cow with clinical mastitis and was characterised by confirmed ability to spread among cows within a herd. Linear regression coefficients were calculated for associations between milk constituents and bacterial counts in whole milk as dependent variables. The comparison of bacterial growth between whole milk, cell reduced, skim and whey fractions was determined by a one-way analysis of variance (ANOVA). The results of the present study showed that the growth of mastitis-causing S. aureus was less stimulated by whole milk samples and their individual fractions in comparison to the nutrient microbiological medium. The strongest inhibition of bacterial growth was observed for whey fraction. Lactoferrin was the only protein causing a slight decrease in the growth of S. aureus. It was concluded that, depending on its growth medium and antimicrobial properties, milk may be among the factors of key importance for the incidence of this disease among individual cows.

Effect of Gluconacetobacter xylinus cultivation conditions on the selected properties of bacterial cellulose

Abstract The aim of the study was to analyze the changes in the parameters of bacterial cultures and bacterial cellulose (BC) synthesized by four reference strains of Gluconacetobacter xylinus during 31-day cultivation in stationary conditions. The study showed that the most visible changes in the analyzed parameters of BC, regardless of the bacterial strain used for their synthesis, were observed in the first 10–14 days of the experiment. It was also revealed, that among parameters showing dependence associated with the particular bacterial strain were the rate and period of BC synthesis, the growth rate of bacteria anchored to the cellulose fibrils, the capacity to absorb water and the water release rate. The results presented in this work may be useful in the selection of optimum culturing conditions and period from the point of view of good efficiency of the cellulose synthesis process.