Enhancement of Inhibition of the Pseudomonas sp. Biofilm Formation on Bacterial Cellulose-Based Wound Dressing by the Combined Action of Alginate Lyase and Gentamicin.

Bacterial biofilms generally contribute to chronic infections, including wound infections. Due to the antibiotic resistance mechanisms protecting bacteria living in the biofilm, they are a serious problem in the wound healing process. To accelerate the wound healing process and avoid bacterial infection, it is necessary to select the appropriate dressing material. In this study, the promising therapeutic properties of alginate lyase (AlgL) immobilised on BC membranes for protecting wounds from Pseudomonas aeruginosa infection were investigated. The AlgL was immobilised on never dried BC pellicles via physical adsorption. The maximum adsorption capacity of AlgL was 6.0 mg/g of dry BC, and the equilibrium was reached after 2 h. The adsorption kinetics was studied, and it has been proven that the adsorption was consistent with Langmuir isotherm. In addition, the impact of enzyme immobilisation on bacterial biofilm stability and the effect of simultaneous immobilisation of AlgL and gentamicin on the viability of bacterial cells was investigated. The obtained results showed that the AlgL immobilisation significantly reduced the amount of polysaccharides component of the P. aeruginosa biofilm. Moreover, the biofilm disruption by AlgL immobilised on BC membranes exhibited synergism with the gentamicin, resulting in 86.5% more dead P. aeruginosa PAO-1 cells.

The Simple Method of Preparation of Highly Carboxylated Bacterial Cellulose with Ni- and Mg-Ferrite-Based Versatile Magnetic Carrier for Enzyme Immobilization.

The bacterial cellulose (BC) is a versatile biopolymer of microbial origin characterized by high purity and unusual water and material properties. However, the native BC contains a low number of functional groups, which significantly limits its further application. The main goal of its effective modification is to use methods that allow the unusual properties of BC to be retained and the desired functional group to be efficiently introduced. In the present study, the new magnetic carrier based on functionalized citric acid (CA) bacterial cellulose was developed and tested to support critical industrial enzymes such as lipase B from Candida antarctica and phospholipase A from Aspergillus oryzae. The applied method allowed BC to be effectively modified by citric acid and a sufficient number of carboxylic groups to be introduced, up to 3.6 mmol of COOH per gram of dry mass of the prepared carrier. The DSC and TGA analyses revealed carrier stability at operational temperatures in the range of 20 °C to 100 °C and substantially influenced the amount of the introduced carboxyl groups on carrier properties. Both enzymes' immobilization significantly improves their thermal stability at 60 °C without a significant thermal and pH optima effect. The analyzed enzymes showed good operational stability with a significant residual activity after ten cycles of repeated uses. The new magnetic carrier based on highly carboxylated bacterial cellulose has a high application capability as matrix for immobilization the various enzymes of industrial interest.

Rotating Magnetic Field Increases ß-Lactam Antibiotic Susceptibility of Methicillin-Resistant Staphylococcus aureus Strains.

Methicillin-resistant strains of Staphylococcus aureus (MRSA) have developed resistance to most ß-lactam antibiotics and have become a global health issue. In this work, we analyzed the impact of a rotating magnetic field (RMF) of well-defined and strictly controlled characteristics coupled with ß-lactam antibiotics against a total of 28 methicillin-resistant and sensitive S. aureus strains. The results indicate that the application of RMF combined with ß-lactam antibiotics correlated with favorable changes in growth inhibition zones or in minimal inhibitory concentrations of the antibiotics compared to controls unexposed to RMF. Fluorescence microscopy indicated a drop in the relative number of cells with intact cell walls after exposure to RMF. These findings were additionally supported by the use of SEM and TEM microscopy, which revealed morphological alterations of RMF-exposed cells manifested by change of shape, drop in cell wall density and cytoplasm condensation. The obtained results indicate that the originally limited impact of ß-lactam antibiotics in MRSA is boosted by the disturbances caused by RMF in the bacterial cell walls. Taking into account the high clinical need for new therapeutic options, effective against MRSA, the data presented in this study have high developmental potential and could serve as a basis for new treatment options for MRSA infections.

Effect of AuNPs and AgNPs on the Antioxidant System and Antioxidant Activity of Lavender (Lavandula angustifolia Mill.) from In Vitro Cultures.

The aim of this study was to determine the effect of gold and silver nanoparticles on the activity of antioxidant enzymes (ascorbate peroxidase (APX), superoxide dismutase (SOD), guaiacol peroxidase (POX), and catalase (CAT)), the free radical scavenging capacity, and the total polyphenol capacity of lavender (Lavandula angustifolia Mill.) cultivar "Munstead" propagated in vitro. In the experiment, fragments of lavender plants were cultivated in vitro on medium with the addition of 1, 2, 5, 10, 20, and 50 mg∙dm-3 of AgNPs or AuNPs (particle sizes 24.2 ± 2.4 and 27.5 ± 4.8 nm, respectively). It was found that the nanoparticles increase the activity of the antioxidant enzymes APX and SOD; however, the reaction depends on the NP concentration. The highest APX activity is found in plants propagated on media with 2 and 5 mg∙dm-3 of AgNPs. AuNPs significantly increase the APX activity when added to media with a concentration of 10 mg∙dm-3. The highest SOD activity is recorded at 2 and 5 mg∙dm-3 AgNP and AuNP concentrations. The addition of higher concentrations of nanoparticles to culture media results in a decrease in the APX and SOD activity. The addition of AuNPs to culture media at concentrations from 2 to 50 mg∙dm-3 increases the POX activity in comparison to its activity when AgNPs are added to the culture media. No significant influence of NPs on the increase in CAT activity was demonstrated. AgNPs and AuNPs increased the free radical scavenging capacity (ABTS•+). The addition of NPs at concentrations of 2 and 5 mg∙dm-3 increased the production of polyphenols; however, in lower concentrations it decreased their content in lavender tissues.

Free and Immobilized Lecitase™ Ultra as the Biocatalyst in the Kinetic Resolution of (E)-4-Arylbut-3-en-2-yl Esters.

The influence of buffer type, co-solvent type, and acyl chain length was investigated for the enantioselective hydrolysis of racemic 4-arylbut-3-en-2-yl esters using Lecitase™ Ultra (LU). Immobilized preparations of the Lecitase™ Ultra enzyme had significantly higher activity and enantioselectivity than the free enzyme, particularly for 4-phenylbut-3-en-2-yl butyrate as the substrate. Moreover, the kinetic resolution with the immobilized enzyme was achieved in a much shorter time (24-48 h). Lecitase™ Ultra, immobilized on cyanogen bromide-activated agarose, was particularly effective, producing, after 24 h of reaction time in phosphate buffer (pH 7.2) with acetone as co-solvent, both (R)-alcohols and unreacted (S)-esters with good to excellent enantiomeric excesses (ee 90-99%). These conditions and enzyme were also suitable for the kinetic separation of racemic (E)-4-phenylbut-3-en-2-yl butyrate analogs containing methyl substituents on the benzene ring (4b,4c), but they did not show any enantioselectivity toward (E)-4-(4'-methoxyphenyl)but-3-en-2-yl butyrate (4d).

A Low Glycemic Index Decreases Inflammation by Increasing the Concentration of Uric Acid and the Activity of Glutathione Peroxidase (GPx3) in Patients with Polycystic Ovary Syndrome (PCOS).

INTRODUCTION: According to a review of the literature, there is a lack of data on the mechanisms that participate in the suppression of inflammation that accompanies polycystic ovary syndrome (PCOS). Additionally, the changes in oxidative status resulting from a low-calorie diet have not been studied in a group of women with PCOS, and the oxidation and reduction processes associated with PCOS have not been explained. MATERIAL AND METHODS: The study involved 49 women who were diagnosed with PCOS according to Rotterdam's criteria, and 24 women voluntarily agreed to a three-month dietary intervention. The dietary intervention was carried out for 3 months. Glutathione peroxidase (GPx3) activity, the Ferric reducing ability of plasma, and uric acid concentration were measured spectrophotometrically both before and after the intervention. Statistical analysis was performed with the Statistica 10.0 software package, and a Pearson's correlation matrix was generated. RESULTS: A lower concentration of GPx3 was observed in women with PCOS (before the dietetic intervention began) compared with the GPx3 levels in healthy women. A relationship was shown between GPx3 levels and the concentration of prolactin, insulin on fasting, and triglycerides. After the dietary intervention, increases in uric acid and GPx3 activity were noted, as well as numerous relationships between anthropometric and biochemical parameters. The ferric reducing/antioxidant power did not change significantly. CONCLUSIONS: Inhibiting the effect of prolactin (by the level of reactive oxygen species) on the activity of GPx3 could be a starting point for the increase in antioxidative stress and the development of the inflammatory state associated with PCOS pathophysiology. Following a low-calorie diet with a lower glycemic index is proposed to silence inflammation by increasing the concentration of uric acid. During GPx3 mobilization, women with PCOS have a higher demand for selenium, and its deficiencies may contribute to disordered thyroid hormone synthesis. The three-month dietary intervention did not silence redox processes in the examined group of women.

Entrapment of DyP-type peroxidase from Pseudomonas fluorescens Pf-5 into Ca-alginate magnetic beads.

The aim of this study was to investigate the optimal conditions for the immobilization and stabilization of DyP1B dye decolorizing peroxidases type B (DyP1B) from Pseudomonas fluorescens Pf-5 immobilized in Ca-alginate ferromagnetic beads. The immobilized DyP1B was used in the degradation of the Reactive Blue 5 (RB5) synthetic dye. The enzyme was successfully entrapped in a Ca-alginate matrix and showed an encapsulation efficiency of 94%. The concentration of DyP1B (0.8 mg mL-1 ), 2% of alginate and magnetite (10.0 mg mL-1 ) was optimal for immobilization. The immobilized DyP1B showed optimum activity at pH 7.0 and 40 °C compared with pH 5.5 and 30 °C for free peroxidase. Reusability studies showed that after five cycles, the immobilized DyP1B system retained more than 58% of its initial activity. The immobilized DyP1B was able to decolorize RB5 at concentrations of 0.1, 0.05, and 0.01% (w v-1 ) with efficiency rates of about 20, 29, and 45%, respectively. The immobilization of DyP1B in alginate beads with the addition of Fe3 O4 increased its catalytic and applicative potential.

Modified Diet Supplementation With Group B Vitamins Changes Antioxidant Defense Activity Of Brain.

The research aimed at effect of modified diet and supplementation with synthetic B vitamins on brain antioxidant status evaluation. Forty male Wistar rats were divided into 4 groups (n = 10); group I - fed Basic Diet (BD), groups II-IV - Modified Diet (MD), where wheat flour replaced 83.5 % wheat and sucrose replaced 50 % maize. Groups I-II received only water, while group III (MD + AS) was given an aqueous vitamins solution supplementing deficiency from diet modification (MD + Adequate Supplementation: B1-0.94, B2-0.48, B6-0.5, niacin-1.9 mg); group IV (MD + ES) received a solution in order to supplement deficiency resulting from the change in diet and recommended prophylactic dose of vitamins (MD + Excessive Supplementation: B1-3.1, B2-2.3, B6-2.4, niacin-6.65 mg). The experimental phase lasted 6 weeks. Blood serum was examined, to determine glucose and iron concentration; determination of FRAP in plasma, in brain tissue the activities of GST, GPx, CAT, SOD, SH content, and FRAP. It was found that MD and MD + AS did not influence cereal antioxidant status. In brain tissue, MD + ES group exhibited an increased glycaemia (7.49 mmol×l-1 p = 0.038) and increased activity of antioxidant enzymes activity (GST 0,099 U/mg protein p = 0.005; GPx 0,039 U/mg protein p = 0.007). Brain tissue of rats also exhibited larger content of protein bond SH in comparison to BD, MD and MD + AS groups (SH 414.6 mmol/g wet weight p < 0.001; p < 0.005). These findings suggest that, an excessive supplementation with vitamins B and niacin, in sucrose rich diet, may lead to enhancing activity of cellular antioxidant defense in brain tissue of rats.

Increased water content in bacterial cellulose synthesized under rotating magnetic fields.

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.

Multiorgan transplantation from a deceased donor with intravascular diffuse large B-cell lymphoma: transmission of the disease and results of treatment.

This report presents the transplantation of two kidneys and the liver from a deceased donor with suspected autoimmune encephalomeningitis (ADEM). Due to an atypical post-transplantation clinical course, the transplanted kidneys were biopsied and this disclosed diffuse large B-cell (DLBC) lymphoma of the intravascular type in each kidney. The same malignancy was found in the postmortem donor brain examination. The renal allografts from the two recipients were removed: despite every effort, one patient died, while chemotherapy was successful in the second. No malignancy was observed in the liver transplant recipient, who received prophylactic chemotherapy. These cases highlight the occasional failure of organ donor disease screening and the consequent unforeseen complications.

Wild boar (Sus scrofa) as a bioindicator of organochlorine compound contamination in terrestrial ecosystems of West Pomerania Province, NW Poland.

The aim of this study was to detect the presence and determine the residue levels of DDT, lindane, endrin and polychlorinated biphenyls (PCBs) in the liver of wild boars from the area of West Pomerania, NW Poland; to determine the activity of glutathione S-transferase (GST) as a biomarker of biological response and to assess the toxicological risk for consumers of the wild boar offal. The presence of pesticide residues and PCBs was found in all examined liver samples. The highest concentration was observed for endrin, and then, the descending order was PCBs >DDTs >lindane >dl-PCBs. The mean hepatic concentrations of endrin, PCBs, DDTs and lindane were 117.28, 78.59, 67.95 and 7.24 ng/g lipid weight, respectively. Among the dioxin-like PCB congeners, 118 and 156 were dominant in liver samples. The mean toxic equivalent (TEQ) level calculated for dl-PCBs was 2.10 ± 1.11 pg WHO-PCB-TEQ/g. There was a statistically significant (p < 0.05) negative correlation between the concentration of lindane, DDTs and PCBs (as a sum of indicator congeners) in the liver and in the activity of GST. However, GST activities showed no significant correlation with any of the dl-PCBs. In five boar liver samples, the levels of certain organochlorine compounds exceeded the maximum residue levels (MRLs). In one sample, the MRLs were exceeded simultaneously for PCBs, endrin and DDTs and in another one-for endrin and DDTs. In the remaining three samples, only PCB levels were exceeded.

Systemic analysis of a manufacturing process based on a small scale bakery.

The main aim of the article is to present two new innovative concepts of reliability of a functioning manufacturing system in the process of making bread in small-scale bakeries. Reliability is understood as one of the representations of an operator acting on specific streams in time to to t. One of these represents the global reliability of a system as a function of parallel action of all the streams of the system in time to to t and is denoted as Pg (t). The second representation of reliability is a scalar value, Pss It shows a new function of global reliability of a manufacturing process as a product of system stream reliability. In order to plot the flow of the manufacturing process's global reliability function, we need to perform detailed calculations, computations, and analysis of the differences of individual values in real time, as well as plan an algorithm of the flow of system streams. This needs a lot of effort, translating however, to a detailed picture of the process. In the analysed example, measurements and research revealed an important increase of the value of reliability in a transition from a traditional to a robotised bakery. The article also presents a new concept of the reliability of a technological process, based on the analysis of relations of elements of the following streams: energy, matter, information, time, and finances. It shows the method of specifying streams and the method for defining the reliability of important and supportive relations. Important relations between stream elements are defined as having the reliability value of one in time. Supportive relations bear the reliability within a continuum between zero and one. Important relations are designated based on research, experience, and knowledge. Stream systemic reliability Pss is a scalar value, i.e. a number from the continuum between zero and one. The Pss value characterises failure-free operation of the whole system. Its average value in the normative time tn expresses the efficiency of the manufacturing system. The value Pss is a quotient of the number of important relation and the sum of important and supportive relations. The formula for Pss shows the method of optimising the process through the increasing of the number of important relations between the input stream components. The concept has been applied to study the efficiency of operation of a small-scale bakery. Systemic analysis of a bakery allows for important increase in the reliability of baking bread if robotisation has been implemented. The concept of systemic-stream reliability Pss may be applied to analyse the efficiency of any technological process and optimisation of any manufacturing process.

Evaluation of new L-amino acids triethanolammonium salts usability for controlling protease activity.

Twenty new triethanolammonium amino acid salts (TEA AA) have been prepared from triethanolammonium hydroxide and L-amino acids. The physicochemical properties of TEA AA depended on the applied amino acid. Five of the synthesised salts, i.e. mono- and bis-salts of L-glutamic acid, L-aspartic acid, and TEA salt of l-glutamine were solids with melting points between 127.32 °C to 171.51 °C. The other TEA AA exhibited glass transition temperatures from -68.45 °C for TEA Ser to -6.27 °C for TEA Trp and were assigned as amino acid ionic liquids (AAILs). The TEA His was characterised by the highest thermal stability, with an average temperature of 5 % weight loss at 186.4 °C, whereas the lowest stability was determined for TEA Asp (107.5 °C). The developed salts were tested as reaction medium additives for proteolytic enzymes (papain, subtilisin, bromelain). Most AAILs showed an inhibitory effect on tested proteases but with different mechanisms related to the enzyme substrate specificity and structural diversity. The TEA Ser was the most effective competitive inhibitor (Ki = 0.24 10-4 mol/L) for bromelain, while TEA Val uncompetitive inhibitor for papain (Ki = 0.25 10-4 mol/L). The developed TEA AA salts exhibit potential as enzyme-controlling agents for use in industrial processes.

Selenium and mercury concentrations, Se:Hg molar ratios and their effect on the antioxidant system in wild mammals.

The aim of this study was to a) evaluate the concentration of Se and Hg and their relationship in the tissues of 4 species of wild mammals, including Se:Hg molar ratios, and b) evaluate the effect of the analysed elements and their mutual proportions expressed as Se:Hg molar ratio, on the antioxidant system in the tissues of the tested animals. The study was performed on 31 animals belonging to four species: wild boar, red fox, roe deer, brown hare. Determination of Hg in liver, kidney and muscle of animals was performed using an AMA 254 mercury analyser. Total Se concentrations were determined using the spectrofluorometric method. In omnivores demonstrated higher Se concentrations in all analysed organs compared to the herbivores. The highest concentration of Hg was found in the kidneys of the tested animals, and the lowest in the muscles. High and moderate correlation between Se and Hg was observed in the liver of omnivorous, while in herbivores this correlation was weak. In all analysed samples, the Se:Hg molar ratios were above 1 (min: liver 5.9, max: kidney 110). Generally, the highest Se:Hg ratio values were found in kidney and the lowest in liver of tested animals. No significant correlation was found between GPx, GST and SOD activity and Se or Hg concentration in analysed organs. But it was observed that Se:Hg molar was negatively correlated with CAT activity in the most samples. The obtained results may suggest that omnivorous animals demonstrate greater Hg sequestration in the liver than herbivores, which has been proposed as one of the mechanisms of Se antagonistic action towards Hg. The ratio between Se and Hg, rather than the concentration of these elements in organs, affected the antioxidant status in the animal organism, specifically the CAT activity.

Argon plasma-modified bacterial cellulose filters for protection against respiratory pathogens.

In this work, we present novel, sustainable filters based on bacterial cellulose (BC) functionalized with low-pressure argon plasma (LPP-Ar). The "green" production process involved BC biosynthesis by Komagataeibacter xylinus, followed by simple purification, homogenization, lyophilization, and finally LPP-Ar treatment. The obtained LPP-Ar-functionalized BC-based material (LPP-Ar-BC-bM) showed excellent antimicrobial and antiviral properties against both Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria, and an enveloped bacteriophage phage Φ6, with no cytotoxicity versus murine fibroblasts in vitro. Further, filters consisting of three layers of LPP-Ar-BC-bM had >99 % bacterial and viral filtration efficiency, while maintaining sufficiently low airflow resistance (6 mbar at an airflow of 95 L/min). Finally, as a proof-of-concept, we were able to prepare 80 masks with LPP-Ar-BC-bM filter and ~85 % of volunteer medical staff assessed them as "good" or "very good" in terms of comfort. We conclude that our novel sustainable, biobased, biodegradable filters are suitable for respiratory personal protective equipment (PPE), such as surgical masks and respirators.

Acceleration of lactose hydrolysis using beta-galactosidase and deep eutectic solvents.

A series of new deep eutectic solvents based on choline levulinate and various hydrogen bond donors were prepared and characterized by1H NMR, FT-IR, TG, and DSC. In particular, their physicochemical properties (density, viscosity, conductivity, and thermal stability) were determined and their usability as an enhancing additive to the enzymatic reaction mixture, for the enzyme was checked. It has been shown, that prepared DES, exhibits low viscosity (at 40 °C within the range 0.1-0.8 Pa·s), high thermal stability (in almost all cases above 150 °C), and density within the range 1.1-1.17 g cm-3. Also, it has been shown, that obtained mixtures can stabilize the enzymes, and positively influence on its activity. The addition of up to 15% (v/v) of DES mixture composed of choline levulinate: ethylene glycol, enhanced more than threefold lactose hydrolysis yield by ß-galactosidase. The present study shows the relevance of the newly designed DES series for improving enzymes properties with the potential to apply in the effective conversion of food processing origin substrates.

An effect of choline lactate based low transition temperature mixtures on the lipase catalytic properties.

A new series of low transition temperature mixures (LTTM) based on choline lactate quaternary ammonium salt and various hydrogen bond donors was prepared and characterized towards their physicochemical properties and usability as an enzymatic reaction mixture for lipase-catalyzed transesterification reactions. Studies of low transition temperature mixtures have shown a long-term stabilizing effect for lipase as well as a positive influence on lipase thermal stability. In the case of Ch[Lac]:Gly: EthGly increasing the stability of lipase by 8 °C (up to 55.2 °C) compared to the control sample. Conducted transesterification reactions were characterized by high yields - up to 98% - and high purity of the obtained products.

Silicone polyether surfactant enhances bacterial cellulose synthesis and water holding capacity.

The versatility and unique properties of bacterial cellulose (BC) motivate research into enhancing its synthesis. Here a silicone polyether surfactant (SPS) was synthesized and tested as a non-nutritional additive to the cultivation media of Komagataeibacter xylinus. The addition of SPS to the Hestrin-Schramm (HS) medium resulted in a concentration-dependent decrease in surface tension from 59.57 ± 0.37 mN/m to 30.05 ± 0.41 mN/m (for 0.1% addition) that was correlated with an increased yield of BC, up to 37% wet mass for surfactant concentration close to its critical micelle concentration (0.008%). Physicochemical characterization of bacterial cellulose obtained in presence of SPS, showed that surfactant is not incorporated into BC structure and has a moderate effect on its crystallinity, thermal stability. Moreover, the water holding capacity was enhanced by over 40%. Importantly, obtained BC did not affect L929 murine fibroblast cell viability. We conclude that SPS provides an eco-friendly approach to increasing BC yield in static culture, enabling more widespread industrial and biomedical applications.

[Legal and medico-legal assessment of medical errors in obstetrics]. / Prawna i sadowo-lekarska ocena bledów medycznych w poloznictwie.

The authors review the doctrine of criminal law and the jurisdiction of the Supreme Court in search of a starting point for the legal protection of human life and health. In cases of medical errors in obstetrics concerning a fetus, an act of a perpetrator can be classified as manslaughter or exposure to direct danger of loss of life or great bodily injury depending on whether the fetus is recognized as "a human being". The authors criticize the doctrinal criteria of the beginning of legal protection: spatial, physiological, obstetric, and developmental, pointing to the possibilities of decriminalization of medical errors of omission. A solution to this situation is the presented evolution of the jurisdiction of the Supreme Court. It moves towards establishing the beginning of legal and criminal protection of life and health at the "moment of emergence of objective grounds to necessitate delivery".

Preparation of Komagataeibacter xylinus Inoculum for Bacterial Cellulose Biosynthesis Using Magnetically Assisted External-Loop Airlift Bioreactor.

The aim of this study was to demonstrate the applicability of a novel magnetically assisted external-loop airlift bioreactor (EL-ALB), equipped with rotating magnetic field (RMF) generators for the preparation of Komagataeibacterxylinus inoculum during three-cycle repeated fed-batch cultures, further used for bacterial cellulose (BC) production. The fermentation carried out in the RMF-assisted EL-ALB allowed to obtain an inoculum of more than 200× higher cellular density compared to classical methods of inoculum preparation. The inoculum obtained in the RMF-assisted EL-ALB was characterized by a high and stable metabolic activity during repeated batch fermentation process. The application of the RMF-assisted EL-ALB for K. xylinus inoculum production did not induce the formation of cellulose-deficient mutants. It was also confirmed that the ability of K. xylinus to produce BC was at the same level (7.26 g/L of dry mass), regardless of inoculum age. Additionally, the BC obtained from the inoculum produced in the RMF-assisted EL-ALB was characterized by reproducible water-related properties, mechanical strength, nano-fibrillar structure and total crystallinity index. The lack of any negative impact of inoculum preparation method using RMF-assisted EL-ALB on BC properties is of paramount value for its future applications, including use as a biomaterial in tissue engineering, wound healing, and drug delivery, where especially BC liquid capacity, nanostructure, crystallinity, and mechanical properties play essential roles.