Presence Of Bacterial Cells Research Articles

Detection and diagnosis of bacterial pathogens in urine using laser-induced breakdown spectroscopy

The presence of bacterial cells from three species has been detected in clinical specimens of human urine using laser-induced breakdown spectroscopy (LIBS) by using a partial least squares discriminant analysis (PLS-DA) of 360 spectra obtained from 12 specimens of infected urine and 239 spectra obtained from eight specimens of sterile urine. Nominally sterile urine specimens obtained from four patients at a local hospital after being screened negative for the presence of bacterial pathogens were spiked with known aliquots of Escherichia coli, Staphylococcus aureus, and Enterobacter cloacae to simulate clinical urinary tract infections. Fifteen emission line intensities measured from the LIBS spectra and 92 ratios of those line intensities were used as 107 independent variables in the PLS-DA for discrimination between bacteria-containing specimens and sterile specimens. The PLS-DA models possessed a 98.3% sensitivity and a 97.9% specificity for the detection of pathogenic cells in urine when single-shot LIBS spectra were tested. To increase the signal to noise ratio, thirty spectra acquired from a single specimen were also averaged together and the averaged spectra were used to construct a model. When each averaged spectrum was withheld from the model individually for testing, the diagnostic test possessed a 100% sensitivity and a 100% specificity for the detection of bacterial cells in urine, although the number of test spectra was necessarily reduced.The entire LIBS spectrum from 200 nm – 590 nm was input into an artificial neural network analysis with principal component analysis pre-processing (PCA-ANN) to diagnose the bacterial species once detected. This PCA-ANN test possessed an overall sensitivity of 97.2%, an overall specificity of 98.6%, and an overall classification accuracy of 97.9% when using 80% of the data to build a model and withholding 20% for cross-validation testing. The PCA-ANN was also performed on each of the 12 bacteria-containing filters individually, using the other 11 filters to build the model. The average sensitivity of this test, calculated by averaging the sensitivities measured for each of the three bacterial species, was 70.9% and the average specificity was 85.5%. Based on these results, the average classification accuracy for the test when used to discriminate between the three microorganisms was 80.6%. These results indicate the potential usefulness of LIBS for rapidly detecting and possibly diagnosing urinary tract infections in a clinical setting.

Susceptibility of the cultured Amazonian fish, Colossoma macropomum, to experimental infection with Aeromonas species from ornamental fish

The global ornamental fish trade carries important risk factors for spreading pathogens between different countries and regions, not only for ornamental fish but also for cultured fish and even other animal species. In the current study, we reported the capacity of Aeromonas veronii and A. hydrophila isolated from ornamental fish to experimentally infect the reared Amazonian fish Colossoma macropomum. For this, those bacteria were identified, and a primary characterization was performed. Fish were inoculated with 0.1 mL of increasing concentrations of A. hydrophila or A. veronii (C1 = 1 × 102; C2 = 1.8 × 104; C3 = 2.1 × 106; C4 = 2.4 × 108 bacterial cells per mL) in the coelomic cavity. In the control group, fish received the same volume of sterile saline solution (0.9 %). Fish presented petechiae, skin suffusions, and mortality rates up to 100 % according to the inoculum concentration. Histopathologically, fish presented necrosis with karyolysis, loss of the cytoplasmic delimitation of cells of the renal tubules and hepatocytes, hemorrhage, cellular edema, and the presence of bacterial cells. The LD50–96h of A. veronii on C. macropomum was estimated at 2.4 × 106 CFU mL−1 and of A. hydrophila at 1.408 × 105 CFU mL−1. The results demonstrated that it is possible that Aeromonas species isolated from ornamental fish affect C. macropomum, causing similar clinical signs and lesions. This shows the importance of promoting risk control measures worldwide regarding the trade of ornamental fish.

Asymptomatic bacteriuria in pregnant Nigerian women at the Lagos University Teaching Hospital, south west Nigeria

Background: Dipstick urinalysis is a handy and cost effective method for the early detection of asymptomatic bacteriuria. Asymptomatic bacteriuria occurs in 2-10% of pregnancies and has been associated with low birth weight and preterm delivery in pregnant women if untreated. Objective: To determine the incidence of asymptomatic bacteriuria in pregnancy, the distribution of infecting organisms and the antibiotic sensitivity pattern in pregnant Nigerian women attending the antenatal clinic of the Lagos University Teaching Hospital, Idi- Araba, Lagos. Methodology: This was a prospective study. The study population were 201 consecutive asymptomatic pregnant women, who attended the antenatal clinic of LUTH for booking between 1st July and 31st December, 2021. Two consecutive urine samples were collected and analysed for microscopy, culture and sensitivity and urinalysis. The outcomes of interest were prevalence of asymptomatic bacteriuria at LUTH, bacterial isolates, and their antibiotic sensitivity pattern. Results: The mean age of the population studied was 31years ± 4.56SD. The mean gestational age was 23.8 ± 5.39 SD with a range of 11-34 weeks; and majority of the participants (65.7%) were recruited at gestational ages between 14 and 26 weeks. The prevalence of asymptomatic bacteriuria in this study was 13.9%. The commonest isolated organism was klebsiella (46.4%) followed by Escherichia coli (21.4%). Ceftazidime, ofloxacin, and gentamycin showed good overall sensitivity with 92.9%, 89.3%, and 67.9%% respectively. From the results of urinalysis, nitrites showed a sensitivity of 48.3% and specificity of 98.2% with a positive predictive value of 83.3% while presence of bacterial cells on urine microscopy showed sensitivity of 31.2%, specificity of 85% and a positive predictive value of 27%. Conclusion: The incidence of asymptomatic bacteriuria in this study was considerably high and therefore, there may be need to consider introduction of routine urine culture at booking or in the second trimester.

A novel combination of immobilized Enterococcus casseliflavus sp. nov. with silver nanoparticles into a reusable matrix of Ca-Alg beads as a new strategy for biotreatment of Disperse Blue 183: Insights into metabolic characterization, biotoxicity, and mutagenic properties

Recent advances in immobilized biologic systems for decolorizing azo dyes are gaining great attention due to microorganisms like bacteria and nanoparticles that could stimulate decolorization. Enhanced decolorization performance was observed in this study, indicating the great potential of the immobilized complex of bacterial cells and AgNPs as an alternative to the traditional biological processes to improve the performance of biological systems. The biodegradation and decolorization of Disperse Blue183 (DB 183) were investigated utilizing a novel combination of Enterococcus casseliflavus strain A2 mediated by silver nanoparticles synthesized by Marinospirillum alkaliphilum strain N in three different conditions. Ⅰ: free bacterial strain A2 (100% dye removal in 72 h), Ⅱ: immobilized bacterial strain A2 in Ca-Alg beads (100% dye removal in 15 h), and Ⅲ: immobilized bacterial strain A2 with silver nanoparticles (AgNPs) as support in Ca-Alg beads (100% dye removal in 9 h). The presence of bacterial cells and nanoparticles in Ca-Alg beads was assessed and proved by scanning electron microscope (SEM) and X-ray energy diffraction (EDX) analysis. Moreover, DB 183 and its decolorization metabolites were evaluated by applying UV–Vis, infrared spectroscopy (FTIR), and GC/MS, and the results showed that the dye was degraded. The antimicrobial effect, brine shrimp toxicity (BST) test, and mutagenicity assay in the presence and absence of metabolic activation (+S9/-S9) were run to assess DB 183 and metabolite obtained from biodegradation. The antimicrobial activity of DB 183 disappeared after treatment. Further, the results of the BST test determined that the dye has moderate biotoxicity (LC50:0.064 mg/mL), and the after-treatment product was not toxic. According to the Ames test, DB 183 had mutagenicity effect (69–84%), and the metabolic activation increased the mutagenicity of the dye) 12–25%). However, the percentage mutagenicity of decolorization products decreased, ranging from 50 to 80% without activation (-S9) and 83–96% in present activation (+S9). This work used the immobilized bacterial cells and AgNPs Ca-Alg gel beads for the first time to introduce this kind of system as a suitable technique for rapid decolorization. Using this application enables a remarkable reduction in the time dedicated to the bioremediation of dyeing wastewater.

Photodynamic inactivation of Staphylococcus aureus in the presence of aggregation-prone photosensitizers based on BODIPY used at submicromolar concentrations.

Two new brominated BODIPYs (1 and 2) bearing amino acid-based chains (l-valine for 1, and dimethyl-l-lysine for 2) were synthesized and characterized. In organic solvents, 1 and 2 were fully soluble and showed the photophysical properties expected for brominated BODIPY dyes, including efficient generation of singlet oxygen (1O2), upon irradiation. In contrast, in aqueous media, both compounds were prone to aggregation and the photo-induced generation of 1O2 was halted. Despite the lack of generation of this reactive species in aqueous media (in cuvette), both 1 and 2 have positive antimicrobial Photodynamic Inactivation (aPDI) effect. The activity against gram-positive Staphylococcus aureus and gram-negative Escherichia coli was determined through the inactivation curves, with a total energy dose of 5.3J/cm2 (white light LED used as an energy source). Compound 2 was highly active against both gram-positive and gram-negative bacteria (3 log CFU/mL reduction was obtained at 0.16μM for S. aureus and 2.5-5.0μM for E. coli), whereas 1 was less effective to kill S. aureus (3 log CFU/mL at 0.32μM) and ineffective for E. coli. The higher efficiency of 2, as compared to 1, to reduce the population of bacteria, can reside in the presence of a protonatable residue in 2, allowing a more effective interaction of this molecule with the cell walls of the microorganisms. In order to explain the lack of reactivity in pure aqueous media (in cuvette) and the contrasting good activity in the presence of bacterial cells it can be hypothesized that upon interaction with the walls of the microorganisms, the aggregated photosensitizers suffer a disaggregation process restoring the ability to generate 1O2, and hence leading to efficient photodynamic activity against these pathogenic microorganisms, in agreement with the similar effect observed recently for porphyrinoid photosensitizers.

Rapid Detection of Gram-Positive and -Negative Bacteria in Water Samples Using Mannan-Binding Lectin-Based Visual Biosensor.

Waterborne bacterial infection is a health threat worldwide, making accurate and timely bacteria detection crucial to prevent waterborne disease outbreaks. Inspired by the intrinsic capability of mannan-binding lectin (MBL) in recognizing the pathogen-associated molecular patterns (PAMPs), a visual biosensor is developed here for the on-site detection of both Gram-positive and -negative bacteria. The biosensor was synthesized by immobilization of the MBL protein onto the blue carboxyl-functionalized polystyrene microparticles (PSM), which is then used in a two-step assay to detect bacterial cells in water samples. The first step involved a 20 min incubation following the MBL-PSM and calcium chloride solution addition to the samples. The second step was to add ethanol to the resultant blue mixture and observe the color change with the naked eye after 15 min. The biosensor had a binary (all-or-none) response, which in the presence of bacterial cells kept its blue color, while in their absence the color changed from blue to colorless. Testing the water samples spiked with four Gram-negative bacteria including Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa and two Gram-positive bacteria of Enterococcus faecalis and Staphylococcus aureus showed that the biosensor could detect all tested bacteria with a concentration as low as 101.5 CFU/ml. The performance of biosensor using the water samples from a water treatment plant also confirmed its capability to detect the pathogens in real-life water samples without the need for instrumentation.

Detonation nanodiamonds as enhancers of E. coli photodynamic inactivation by phthalocyanines in a high molarity buffer solution.

Antimicrobial therapy, especially inactivation of multi-antibiotic-resistant strains, requires creating new approaches for drug action and targeted delivery in different environmental conditions. In this work, detonation nanodiamonds (DNDs) were used to deliver polycationic zinc phthalocyanines to E. coli cells. It is shown that in aqueous solutions, zinc phthalocyanines with cholinyl peripheral substituents form complexes with negatively charged DND based on electrostatic interactions. About 40-70 phthalocyanine molecules can bind to a single DND particle, depending on the number of charged groups of the dye molecule. During the complex formation, quenching of phthalocyanine fluorescence and a decrease in its ability to generate reactive oxygen species were observed. In the presence of bacterial cells, phthalocyanine left the complex and induced a photodynamic effect, the magnitude of which depended on the phthalocyanine charge, the molarity of the buffer solution, and the stoichiometry of the phthalocyanine-DND complex. It was found that at physiological values of the ionic strength of the solution, the photodynamic effect of phthalocyanine with a charge of 8+ in combination with a DND is higher than that of the initial phthalocyanine. Thus, nanodiamonds are a promising platform for the delivery of photosensitizers in antimicrobial therapy.

A NOVEL NOX/PHOX-CD38-NAADP-TFEB AXIS IMPORTANT FOR MACROPHAGE ACTIVATION DURING BACTERIAL PHAGOCYTOSIS

ABSTRACT Macrophage activation in the presence of bacterial cells and molecules entails complex programs of gene expression. How such triggers elicit specific gene expression programs is incompletely understood. We previously discovered that TFEB (transcription factor EB) is a key contributor to macrophage activation during bacterial phagocytosis. However, the mechanism linking phagocytosis of bacterial cells to TFEB activation and downstream pro-inflammatory cytokine induction remained unknown. We found that macrophages lacking both TFEB and TFE3 (transcription factor E3) were unable to mount a pro-inflammatory phenotype in response to bacterial infection. The NOX/PHOX (NADPH oxidase)-dependent oxidative burst was required for nuclear translocation of TFEB during phagocytosis of Gram-positive or -negative bacteria, and reactive oxygen species (ROS) were sufficient to trigger TFEB activation in a CD38- and NAADP (nicotinic acid adenine dinucleotide phosphate)-dependent manner. Consistent with the Ca2+-releasing activity of NAADP, intracellular Ca2+ chelation and PPP3/calcineurin inhibition prevented TFEB activation by phagocytosis and ROS (reactive oxygen species), impairing the induction of pro-inflammatory cytokines such as IL6 and TNF/TNFα. Therefore, here we describe a previously unknown pathway that links phagocytosis with macrophage pro-inflammatory polarization via TFEB and related transcription factor TFE3. These findings reveal that activation of TFEB and TFE3 is a key regulatory event for the activation of macrophages, and have important implications for infections, inflammation, cancer, obesity, and atherosclerosis.

The biotechnology approach for sustainable concrete material – a review

Biomineralisation is a process of formation of calcite from supersaturated solution due to the presence of bacterial cells and biochemical activities. In this article, a biological approach towards the development of sustainable construction materials is studied. Evidence of bacterial precipitation of calcite has led several research groups to explore the possibility of adopting this process in the area of construction materials. The goal of this review paper is to highlight the application of biomineralisation in concrete. The effect of a bio-based agent and bacterial solution on various properties of concrete is discussed. Additionally, the factors in the choice of bacteria for the biomineralisation process are also elaborated. This green biotechnology concept is a promising, environmentally safe substitute for conventional and current remediation methods to mitigate environmental problems in multidisciplinary fields.

A study of corrosion processes in a demineralized aqueous medium in the presence of bacterial cells

Our earlier results indicate the feasibility of considering the microbiological contamination of closed and reversible cooling and water supply systems, given the effect of the microbial cells on the corrosion processes taking place there. The results show a possibility of transition metal ions acting as regulators and modifiers of microorganisms’ growth processes under conditions of an osmotic stress and starvation.The aim of the present work is to study the effect of transition d-metal ions (Zn2+ and Cr3+), both separately and in the presence of known antiskalants, oxyethylenediphosphonic acid (OEDFh) and nitrilotrimethylphosphonic acid (NTMFh), on the effectiveness of the corrosion protection and the course of the biological corrosion in a demineralized environment.Microorganisms belonging to different systematic affiliations were selected for the study of the biocorrosion processes: Bacillus sp. and Pseudomonas sp. These microorganisms belonged to different physiological groups of bacteria and had opposite types of life strategies. They differred significantly in their ability to survive under the influence of various factors. They could exist in closed systems of water consumption (with sharp temperature changes, oxygen starvation, a presence of toxic substances used for water conditioning). Bacillius and Pseudomonas strains were cultured on commonly used agar media, after which a cell suspension was prepared. For all bacterial strains used, the seeding dose was 108 cells‧cm-3. It was determined using turbidometric control methods and bacterial density standards. The effect of the microorganisms on the corrosion processes was followed in stationary bioreactors of a volume of 125 cm3. A solution of 0.1 % NaCl was added as a model medium to which corrosion inhibitors (Zn2+, Cr3+, OEDPh and NTMPh) were added at a concentration of 5 mg‧dm-3. At each stage of preparation, the suspension was thoroughly mixed, after which the metal samples were introduced to the reactor in the form of electrodes so that the entire surface of the electrode was completely immersed in the water. The metal samples were made of St 20 carbon steel. Each study of model water with metal plates was of 3 weeks duration. The intensity of the corrosion process was evaluated once a week. The rate of the biological corrosion was determined by the gravimetric method. The inhibition factor (j) and the degree of the corrosion protection (Z, %) of the samples in presence of corrosion inhibitors and bacteria were calculated. The negative value of this indicator indicated a change of the direction of the process, i.e. an activation of the corrosion process.The obtained data show that the presence of microorganisms significantly modifies the anti-corrosion effect of the heavy metal ions. The range offluctuations of the degree of the corrosion protection in presence of bacteria of the genus Pseudomonas is greater than that of the genus Bacillus. The dynamics of the protective effect in presence of both strains is characterized by significant changes (a decrease and a subsequent gradual increase of the degree of the corrosion protection). The highest intensity of the corrosion process is observed during the first week of the interaction in a model environment containing bacterial cells and anticorrosive agents. Then, the direction of the process changed and there was shown a slight protective effect at the 5-15 up to 36 % level. It is also demonstrated that the anti-corrosion compositions containing NTMPh provide a greater increase of the corrosion process rate in presence of microorganisms.The study of the presence of microorganisms on the corrosion proceeding demonstrates their ability to provide a significant effect. The bacteria can activate the corrosion process, which complicates the inhibitors behavior.

Urinary Biomarkers: Diagnostic Tools for Monitoring Athletes' Health Status.

Acute or intense exercise is sometimes related to infections of the urinary tract. It can also lead to incorrect hydration as well as incorrect glomerular filtration due to the presence of high-molecular-weight proteins that cause damage to the kidneys. In this context, our study lays the foundations for the use of a urine test in a team of twelve male basketball players as a means of monitoring numerous biochemical parameters, including pH, specific weight, color, appearance, presence of bacterial cells, presence of squamous cells, leukocytes, erythrocytes, proteins, glucose, ketones, bilirubin, hemoglobin, nitrite, and leukocyte esterase, to prevent and/or treat the onset of pathologies, prescribe personalized treatments for each athlete, and monitor the athletes’ health status.

Probing extracellular reduction mechanisms of Bacillus subtilis and Escherichia coli with nitroaromatic compounds

Redox transformations of organic contaminants by bacterial extracellular polymeric substances (EPS) and the associated electron transfer mechanisms are rarely reported. Here we show that a nitroaromatic compound (1,3-dinitrobenzene) can be readily reduced to 3-hydroxylaminonitrobenzene and 3-nitroaniline in aqueous suspension of common bacteria (E. coli or B. subtilis) or in aqueous dissolved EPS extracted from the bacteria. The loss ratio of 1,3-dinitrobenzene by E. coli was unaffected after knocking out the nfsA gene encoding nitroreductase, but was suppressed by removing EPS attached to cells. In contrast, the loss ratio was enhanced by adding aqueous dissolved EPS to E. coli or B. subtilis suspension. The residual 1,3-dinitrobenzene and products formed after reduction were only presented outside the bacterial cells. Thus, bacterial reduction of 1,3-dinitrobenzene was mediated by nonenzymatic extracellular reduction. This was further corroborated by the observation that the stoichiometric demand of electrons in 1,3-dinitrobenzene reduction was nearly equal to the quantity of electrons donated by bacterial cells in the electrochemical cell experiment. Inhibition on the reduction of 1,3-dinitrobenzene by chemical probes combined with fluorescence detection demonstrated that reducing sugars in EPS might act as electron donors, while cytochromes and some low-molecular weight molecules (flavins and quinones) were involved as electron transfer mediators. Linear relationships were observed between the reduction kinetics and the one-electron reduction potentials for a series of substituted dinitrobenzenes in the presence of bacterial cells or dissolved EPS. Their close linear regression slope values suggest that the extracellular matrix and the exfoliated EPS utilized the same reducing agents (likely hydroquinones and reduced flavins) as terminal electron donors to reduce NACs. These results reveal a previously unrecognized mechanism for nonenzymatic extracellular reduction of NACs by common bacteria. CapsuleThe extracellular matrix of E. coli or B. subtilis supplies both electron donors and electron transfer mediators to efficiently reduce nitroaromatic compounds.

Molecular Mechanisms of Lead Binding to Ferrihydrite-Bacteria Composites: ITC, XAFS, and μ-XRF Investigations.

Binding of Pb(II) to ferrihydrite-Bacillus subtilis composites formed in the presence of bacterial cells were investigated through macroscopic and microscopic techniques. Diffuse layer model (DLM) fitting and isothermal titration calorimetry (ITC) analysis indicated that the hydroxyl group played a key role in Pb(II) sorption onto composites by masking reactive sites, such as carboxyl and phosphoryl groups of bacterial cells. Negative enthalpy (from -39.29 to -57.87 kJ mol-1) and positive entropy (from 135.61 to 193.47 kJ mol-1) of Pb(II) sorption onto composites revealed that inner-sphere complexes formed through exothermic reactions and was driven by both entropy and enthalpy. Spatial distribution of these inner-sphere species at varied Pb(II) loading demonstrated that interactions between Pb(II) and bacterial cells preceded that of mineral components in composites, using microfocus X-ray fluorescence spectroscopy (μ-XRF) maps and microfocus X-ray absorption near edge structure (μ-XANES) spectra. Combined with bulk Pb LIII-edge X-ray absorption fine structure (XAFS) spectrum, we inferred that mononuclear bidentate edge-sharing hydroxyl-Pb complexes, monodentate mononuclear carboxyl-Pb and phosphoryl-Pb complexes predominantly contributed to Pb(II) inner-sphere binding with mineral and bacterial fractions in composites, respectively. The molecular binding mechanisms obtained in this study provide further insight into the sequestration and migration of toxic metals in natural environments.

Investigation of the Influence of Aging on the Lubricity of Metalworking Fluids by Means of Design of Experiment

The influence of complex aging processes in water-miscible metalworking fluids on process performance is of high relevance for the metalworking industry. Because of the highly dynamic interactions in the complex “metalworking fluid” ecosystem, a distinct correlation between the aging process and the performance of the fluid in metalworking processes is hardly possible. Consequences of the aging process on physical, chemical, and biological properties of the fluid include aspects such as the decrease of the pH value, the increase of the droplet size in emulsions, the presence of bacterial cells, or the modification of the metalworking fluid composition. In the presented work, the influences of these aging aspects on the lubricity of metalworking fluids were investigated individually. A test series has been carried out, which was planned with a design of experiments method, to investigate interactions between the aging aspects regarding lubricity. In addition, the results enabled the development of an empirical regression model, which allowed an integrated description of the influence of the relevant aging aspects.

The dialogue between protozoa and bacteria in a microfluidic device.

In nature, protozoa play a major role in controlling bacterial populations. This paper proposes a microfluidic device for the study of protozoa behaviors change due to their chemotactic response in the presence of bacterial cells. A three-channel microfluidic device was designed using a nitrocellulose membrane into which channels were cut using a laser cutter. The membrane was sandwiched between two glass slides; a Euglena suspension was then allowed to flow through the central channel. The two side channels were filled with either, 0.1% peptone as a negative control, or a Listeria suspension respectively. The membrane design prevented direct interaction but allowed Euglena cells to detect Listeria cells as secretions diffused through the nitrocellulose membrane. A significant number of Euglena cells migrated toward the chambers near the bacterial cells, indicating a positive chemotactic response of Euglena toward chemical cues released from Listeria cells. Filtrates collected from Listeria suspension with a series of molecular weight cutoffs (3k, 10k and 100k) were examined in Euglena chemotaxis tests. Euglena cells were attracted to all filtrates collected from the membrane filtration with different molecular weight cutoffs, suggesting small molecules from Listeria might be the chemical cues to attract protozoa. Headspace volatile organic compounds (VOC) released from Listeria were collected, spiked to 0.1% peptone and tested as the chemotactic effectors. It was discovered that the Euglena cells responded quickly to Listeria VOCs including decanal, 3,5- dimethylbenzaldehyde, ethyl acetate, indicating bacterial VOCs were used by Euglena to track the location of bacteria.

Photodynamic efficacy of toluidine blue O against mono species and dual species bacterial biofilm.

The purpose of this study was to investigate how Enterococcus faecalis and Streptococcus mutans behave in mono and dual species biofilm after photodynamic treatment. Antimicrobial photodynamic therapy (aPDT) leads to the generation of reactive oxygen species (ROS) that destroys bacterial cells in presence of a photosensitizer, visible light, and oxygen. We have taken Enterococcus faecalis and Streptococcus mutans as monospecies culture and their dualspecies culture biofilm. Antibacterial effect was evaluated by colony forming unit while antibiofilm action by crystal violet and congored binding assays. We found that dual species biofilm are more resistant than monospecies biofilm and S. mutans shows dominance over E. faecalis in dual species biofilm, it inhibited the growth of E. faecalis in dual species biofilm. Antibiofilm efficacy of TBO also validated that dualspecies show less inhibition than monospecies biofilm this may be due to different EPS constitution in dualspecies biofilm, hence less inhibition was observed in EPS production of dualspecies biofilm than monospecies biofilm. Reactive oxygen species and singlet oxygen yield was found to be light dose dependent and antimicrobial photodynamic efficiency is directly related to the ROS production. We conclude that dual species biofilm shows resistance over monospecies biofilm and S. mutans in dual species inhibits the growth of E. faecalis.

In Situ Real-Time Study on Dynamics of Microbially Induced Calcium Carbonate Precipitation at a Single-Cell Level.

Ureolytic microbially induced calcium carbonate precipitation (MICP) is a promising green technique for addressing a variety of environmental and architectural concerns. However, the dynamics of MICP especially at the microscopic level remains relatively unexplored. In this work, by applying a bacterial tracking technique, the growth dynamics of micrometer-sized calcium carbonate precipitates induced by Sporosarcina pasteurii were studied at a single-cell resolution. The growth of micrometer-scale precipitates and the occurrence and dissolution of many unstable submicrometer calcium carbonate particles were observed in the precipitation process. More interestingly, we observed that micrometer-sized precipitated crystals did not grow on negatively charged cell surfaces nor on other tested polystyrene microspheres with different negatively charged surface modifications, indicating that a negatively charged surface was not a sufficient property for nucleating the growth of precipitates in the MICP process under the conditions used in this study. Our observations imply that the frequently cited model of bacterial cell surfaces as nucleation sites for precipitates during MICP is oversimplified. In addition, additional growth of calcium carbonates was observed on old precipitates collected from previous runs. The presence of bacterial cells was also shown to affect both morphologies and crystalline structures of precipitates, and both calcite and vaterite precipitates were found when cells physically coexisted with precipitates. This study provides new insights into the regulation of MICP through dynamic control of precipitation.

Effect of Acylation on the Antimicrobial Activity of Temporin B Analogues.

New peptides derived from the natural antimicrobial temporin B were obtained. The design, antimicrobial activity, and characterization of the secondary structure of peptides in the presence of bacterial cells is described herein. TB_KKG6K (KKLLPIVKNLLKSLL) has been identified as the most active analogue against Gram-positive and -negative bacteria, compared with natural temporin B (LLPIVGNLLKSLL) and TB_KKG6A (KKLLPIVANLLKSLL). Acylation with hydrophobic moieties generally led to reduced activity; however, acylation at the 6-position of TB_KKG6K led to retained sub-micromolar activity against Staphylococcus epidermidis.

A Static Microfluidic Device for Investigating the Chemotaxis Response to Stable, Non-linear Gradients.

Microfluidic technology allows fast and precise measurement of chemotaxis responses to both attractant and repellent signals. One of the major drawbacks of current microfluidic chemotaxis assays is the presence of bacterial cells within the concentration gradient flow field, which has the potential for flow effects masking the chemotaxis response. This chapter describes a new microfluidic device for producing stable concentration gradients and measuring the response of cells to the gradient without exposing them to any flow. Unlike other methods described in the literature, this method is capable of producing gradients of any shape, almost instantaneously, allowing the measurement of time-dependent response of cells to a variety of signals.

Immunobiological and morphofunctional indicators with dysbacteriosis of the intestine of rabbits

The article presents results of research of dynamics of morphological, biochemical, haematological and immunological parameters when changing the quantity and species of microorganisms of intestinal microbiocenosis of the rabbits. With the dominance of pathogenic bacteria in rabbit intestinal microbiocenoses, the titer of bifidumbacteria was 10-5-10-4, a decrease in the number of lactobacilli, an increase in the number of enterobacteria, a correlation between the colonization index and the degree of adhesion of bacteria was revealed; an increase in the total number of leukocytes, lymphocytes, erythrocytes, hematocrit, alanine aminotransferase level, aspartate aminotransferase, lactate dehydrogenase, total bilirubin, -amylase, albumin, alkaline phosphatase, T-killer level, NK cells, concentration of C-reactive protein. We observed a decrease in the concentration of hemoglobin, monocytes, total protein level, T-lymphocytes, T-helpers, B-lymphocytes, phagocytic index, phagocytic activity of leukocytes, NST-spontaneous, NST-stimulated. Dynamics of morphological indicators was characterized by a disorder of the circulation of organs, a predominance of dystrophic and necrotic processes in the tissues and organs of the cardiovascular, respiratory, digestive, excretory system. Infiltration with mononuclear and segmented leukocytes was noted, diffuse lymphoid proliferation of the mucous membranes of the organs, hyperemia, edema, multiple hemorrhages, accumulation of hemorrhagic exudate in the lumen of the respiratory, digestive and urogenital tracts. Observed catarrhal-hemorrhagic inflammation of the gastrointestinal tract; mucous membrane of the stomach, small and large intestines edematic, hyperemic, with hemorrhages, covered with mucus, fibrinous films, with necrotic areas. The signs of acute diffuse interstitial nephritis, characterized by proliferation of connective tissue cells with a predominance of lipoblast and epithelioid cells, are revealed. In the presence of bacterial cells in the lumen of the tubules, the parenchyma of the cortical and medullar layers was marked by microabscesses. Accidental transformation of the thymus was observed, characterized by signs of a circulatory disorder, a decrease in the area of the cortex, the formation of cysts; signs of diffuse hyperplasia of the spleen with necrobiotic changes in the foci of proliferation, reduction of the area of lymphoid follicles, expansion of the area occupied by the red pulp of follicles, hyperplasia of follicles of white pulp with foci of necrosis. Identified multiple bacterial embolism of blood vessels, developed signs of hemorrhagic Diathesis, watched the development of sign infiltrative itch processes.