Adhesion heterogeneity of individual bacterial cells in an axenic culture studied by atomic force microscopy.

The evaluation of bacterial adhesive properties at a single-cell level is critical for under standing the role of phenotypic heterogeneity in bacterial attachment and community formation. Bacterial population exhibits a wide variety of adhesive properties at the single-cell level, suggesting that bacterial adhesion is a rather complex process and some bacteria are prone to phenotypic heterogeneity. This heterogeneity was more pronounced for Escherichia coli, where two subpopulations were detected. Subpopulations exhibiting higher adhesion forces may be better adapted to colonize a new surface, especially during sudden changes in environmental conditions. Escherichia coli was characterized by a higher adhesion force, a stronger ability to form biofilm and larger heterogeneity index calculated in comparison with Bacillus subtilis. Higher adhesion forces are associated with a more efficient attachment of bacteria observed in an adhesion assay and might provide a basis for successful colonization, survival and multiplications in changing environment. The atomic force microscopy provides a platform for investigation of the adhesion heterogeneity of individual cells within a population, which may be expected to underpin further elucidation of the adaptive significance of phenotypic heterogeneity in a natural environment.

Controlling an Industrial Robot Using a Graphic Tablet in Offline and Online Mode.

The article presents the possibility of using a graphics tablet to control an industrial robot. The paper presents elements of software development for offline and online control of a robot. The program for the graphic tablet and the operator interface was developed in C# language in Visual Studio environment, while the program controlling the industrial robot was developed in RAPID language in the RobotStudio environment. Thanks to the development of a digital twin of the real robotic workstation, tests were carried out on the correct functioning of the application in offline mode (without using the real robot). The obtained results were verified in online mode (on a real production station). The developed computer programmes have a modular structure, which makes it possible to easily adapt them to one's needs. The application allows for changing the parameters of the robot and the parameters of the path drawing. Tests were carried out on the influence of the sampling frequency and the tool diameter on the quality of the reconstructed trajectory of the industrial robot. The results confirmed the correctness of the application. Thanks to the new method of robot programming, it is possible to quickly modify the path by the operator, without the knowledge of robot programming languages. Further research will focus on analyzing the influence of screen resolution and layout scale on the accuracy of trajectory generation.

Selection and Optimization of the Parameters of the Robotized Packaging Process of One Type of Product.

The article presents the results of computer simulations related to the selection and optimization of the parameters of robotic packing process of one type of product. Taking the required performance of the robotic production line as a basis, we proposed its configuration using the RobotStudio environment for offline robot programming and virtual controller technology. Next, a methodology for the validation of the adopted assumptions was developed, based on a wide range of input data and a precise representation of the applicable conditions in the packaging process of one type of product. This methodology included test scenarios repeated an appropriate number of times in order to obtain the result data with the desired reliability and repeatability. The main element of the research was a computer simulation of the station based on the Picking PowerPac package. It was assumed that the products on the technological line are generated pseudo-randomly, thus reflecting the real working conditions. The result of the conducted works is the optimal operating speed of industrial robots and conveyors. The developed methodology allows for multifaceted analyses of the key parameters of the technological process (e.g., the number of active robots and their load, speed of conveyors, and station efficiency). We paid special attention to the occurrence of anomalies, i.e., emergency situations in the form of "halting" the operation of chosen robots and their impact on the obtained quality of the industrial process. As a result of the simulations, numerical values were obtained, maximum efficiency, with regard to maximum overflow of items of 5%, for LB algorithm was equal to 1188 completed containers per hour, with conveyors speeds of 270 mm/s and 165 mm/s. This efficiency was possible at robot speeds R1 = 6450 mm/s, R2 = 7500 mm/s, R3 = 6500 mm/s, R4 = 6375 mm/s, R5 = 5500 mm/s, R6 = 7200 mm/s. The ATC algorithm reached efficiency of 1332 containers per hour with less than 5% overflown items, with conveyor speeds of 310 mm/s and 185 mm/s. This efficiency was possible at robot speeds R1 = 7500 mm/s, R2 = 7500 mm/s, R3 = 7200 mm/s, R4 = 7000 mm/s, R5 = 6450 mm/s, R6 = 6300 mm/s. Tests carried out for emergency situations showed that the LB algorithm does not allow for automatic continuation of the process, while the ATC algorithm assured production efficiency of 94% to 98% of the maximum station efficiency.

Clove Oil (Syzygium aromaticum L.) Activity against Alicyclobacillus acidoterrestris Biofilm on Technical Surfaces.

Acidotermophilic bacteria Alicyclobacillus acidoterrestris is one of the main contaminants in the fruit industry forming biofilms which are difficult to remove from the production line by conventional methods. An alternative approach aims for the use of essential oils to prevent Alicyclobacillus biofilm development. The effect of clove essential oil on A. acidoterrestris biofilms on glass and polyvinyl chloride surfaces under static and agitated culture conditions was investigated by atomic force microscopy and the plate count method. The medium-flow and the type of technical surface significantly influenced A. acidoterrestris biofilm. The PVC was colonized in a greater extent comparing to glass. Clove essential oil in 0.05% (v/v) caused 25.1-65.0% reduction of biofilms on the technical surfaces along with substantial changes in their morphology by a decrease in the biofilm: height, surface roughness, and surface area difference. The oil also induced alteration in individual bacterial cells length and visible increase of their roughness. Clove essential oil seems to release EPS from biofilm and thus induce detachment of bacteria from the surface. Due to anti-A. acidoterrestris biofilm activity, the clove oil may be used in the juice industry to hinder a development of A. acidoterrestris biofilms on production surfaces.

Molecular Mechanisms of Leonurus Cardiaca L. Extract Activity in Prevention of Staphylococcal Endocarditis-Study on in Vitro and ex Vivo Models.

Better understanding the mechanisms of Leonurus cardiaca L. extract (LCE) activity is necessary to prepare recommendations for the use of LCE-based herbal products for preventive/supportive purposes in case of infective endocarditis (IE) and other staphylococcal invasive infections. The aim of the study was to analyze molecular mechanisms of LCE effect on Staphylococcus aureus and blood platelets in the context of their interactions playing a pivotal role in such disorders. Using atomic force microscopy, we demonstrated that adhesion forces of S. aureus were markedly reduced after exposure to LCE at subinhibitory concentrations. The effect resulted from the impact of LCE on S. aureus cell morphology and the composition of phospholipids and fatty acids in bacterial membranes (assessed by HPLC), which modulated their stabilization, hydrophobicity, and charge. Moreover, using FACS we showed also that LCE significantly reduced GP IIb/IIIa expression on blood platelets, thus the disruption of platelet-fibrinogen interactions seems to explain antiplatelet effect of LCE. The obtained results prove the usefulness of LCE in the prevention of S. aureus adhesion, platelet activation, and vegetations development, however, also pointed out the necessity of excluding the cationic antibiotics from the treatment of S. aureus-associated IE and other invasive diseases, when motherwort herb is used simultaneously as an addition to the daily diet.

Effect of ampicillin on adhesive properties of bacteria examined by atomic force microscopy.

Discovery of new antibacterial agents requires the development of novel techniques for bacteria surface characterization after treatment with antibiotics. In this study, we investigate the effect of ampicillin at MICs levels on adhesive properties of Gram-positive and Gram-negative bacteria, using atomic force microscopy (AFM). Our results revealed that the treatment leads to changes of bacterial surface properties, especially cell surface roughness. A nanomechanical alteration of the cells led to an increase of adhesive forces and rupture lengths. Changes in adhesive properties are determined not only by the modification of physicochemical cell properties but also by an increase in roughness, leading to an increase of the contact area with a cantilever tip. We discovered that the contribution of non-specific physicochemical interactions in the bacteria attachment to a substrate is not negligible and was significantly influenced by the presence of antibiotic. Ampicillin caused much greater change in the adhesion properties of Bacillus subtilis than Escherichia coli due to the mode of action of ß-lactam antibiotic. Adhesion measurements may by a new way to investigate subtle changes of the bacterial surface properties caused by antibiotic, especially those targeting the bacterial cell wall. In contrast to nanoindentation assays, they provide information on adhesive properties of the bacteria surface.

Antimicrobial activity of biosilver nanoparticles produced by a novel Streptacidiphilus durhamensis strain.

BACKGROUND/PURPOSE: In this study, an acidophilic actinobacteria strain was used as a novel reducing agent for a single-step synthesis of nanostructure silver particles. We used a Streptacidiphilus durhamensis HGG16n isolate for efficient synthesis of bioactive silver nanoparticles [bio(AgNPs)] in an inexpensive, eco-friendly, and nontoxic manner. The obtained bio(AgNPs) exhibited unique physicochemical and biochemical properties. METHODS: Structural, morphological, and optical properties of the synthesized biocolloids were characterized by spectroscopy, dynamic light scattering, and electron microscopy approaches. The antimicrobial activity was evaluated using the well- and disc-diffusion methods. RESULTS: The obtained crystalline structure and stable biosynthesized silver nanoparticles ranged in size from 8 nm to 48 nm and were mostly spherical in shape. Antimicrobial assays of the silver nanoparticles against pathogenic bacteria showed the highest antimicrobial activity against Pseudomonas aeruginosa, Staphylococcus aureus, and Proteus mirabilis, followed by Escherichia coli, Klebsiella pneumoniae, and Bacillus subtilis. Moreover, the synergistic effect of bio(AgNPs) with various commercially available antibiotics was also evaluated. CONCLUSION: These results provide insight into the development of new antimicrobial agents along with synergistic enhancement of the antibacterial mechanism against clinical bacteria.