Suspensions Of Bacteria Research Articles

Colloidal Structure Dictates Antimicrobial Efficacy in LL-37 Self-Assemblies With Glycerol Monooleate.

The antimicrobial peptide LL-37 is a promising alternative to conventional antibiotics to combat bacteria in suspension and biofilms. Its self-assembly with polar lipids is suggested to improve its potential for therapeutic applications with higher stability against degradation and bioavailability. This study investigates the self-assembly of LL-37 with glyceryl monooleate (GMO), establishing the link between colloidal structure and antimicrobial activity. Small-angle X-ray scattering, dynamic light scattering and cryogenic transmission electron microscopy show structural transformation from dispersions of inverse bicontinuous structure (cubosomes) to multilamellar vesicles and direct rod-like mixed-micelles upon increasing the content of LL-37 in GMO. In vitro assays against planktonic and biofilm cells demonstrate that 128 µg mL-1 of GMO cubosomes have no impact on Pseudomonas aeruginosa. Still, the cubosomes reduce the Staphylococcus aureus planktonic population by ≈ 1-log after 24 h. Cylindrical micelles formed at LL-37/GMO 9/1 and 8/2 with 128 µg mL-1 LL-37 decrease the Pseudomonas aeruginosa population by 6-log. This activity is gradually abolished when LL-37 is encapsulated in vesicles or cubosomes. They also demonstrate low antibiofilm efficacy and promote the biomass of Staphylococcus aureus biofilms. These results highlight the importance of colloidal structure for therapeutic outcomes, providing insights for advanced lipid nanocarrier designs.

Enhanced Diffusion and Non-Gaussian Displacements of Colloids in Quasi-2D Suspensions of Motile Bacteria

In the real world, active agents interact with surrounding passive objects, thus introducing additional degrees of complexity. The relative contributions of far-field hydrodynamic and near-field contact interactions to the anomalous diffusion of passive particles in suspensions of active swimmers remain a subject of ongoing debate. We constructed a quasi-two-dimensional microswimmer–colloid mixed system by taking advantage of Serratia marcescens’ tendency to become trapped at the air–water interface to investigate the origins of the enhanced diffusion and non-Gaussianity of the displacement distributions of passive colloidal tracers. Our findings reveal that the diffusion behavior of colloidal particles exhibits a strong dependence on bacterial density. At moderate densities, the collective dynamics of bacteria dominate the diffusion of tracer particles. In dilute bacterial suspensions, although there are multiple dynamic types present, near-field contact interactions such as collisions play a major role in the enhancement of colloidal transport and the emergence of non-Gaussian displacement distributions characterized by heavy exponential tails in short times. Despite the distinct types of microorganisms and their diverse self-propulsion mechanisms, a generality in the diffusion behavior of passive colloids and their underlying dynamics is observed.

Preparation of Antimicrobial Agents: From Interpolyelectrolyte Complexes to Silver-Containing Metal–Polymer Complexes and Nanocomposites

In order to control pathogenic microorganisms, three polymer compositions were prepared and tested. First, a water-soluble positively charged polycomplex was synthesized via the electrostatic binding of anionic polyacrylic acid to an excess of polyethylenimine to enhance the biocidal activity of the polycation. Second, an aqueous solution of AgNO3 was added to the polycomplex, thus forming a ternary polycation-polyanion-Ag1+ complex with an additional antimicrobial effect. Third, the resulting ternary complex was subjected to UV irradiation, which ensured the conversion of Ag1+ ions into Ag nanoparticles ranging in size mainly from 10 to 20 nm. Aqueous solutions of the polymer compositions were added to suspensions of the Gram-positive bacteria S. aureus and the Gram-negative bacteria P. aeruginosa, with the following main results: (a) Upon the addition of the binary polycomplex, 30% or more of the cells survived after 20 h. (b) The ternary complex killed S. aureus bacteria but was ineffective against P. aeruginosa bacteria. (c) When the ternary complex with Ag nanoparticles was added, the percentage of surviving cells of both types did not exceed 0.03%. The obtained results are valuable for the development of antibacterial formulations.

Microplastic biofilms promote the horizontal transfer of antibiotic resistance genes in estuarine environments

As emerging pollutants, microplastics can aggregate microorganisms on their surfaces and form biofilms, enriching antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). Consequently, microplastic biofilms have become a focal point of research. Horizontal gene transfer is one of the primary mechanisms by which bacteria acquire antibiotic resistance, with much of the research focusing on suspended bacteria. However, microplastic biofilms, as hotspots for horizontal gene transfer, also merit significant investigation. This study primarily explored and compared the frequency of ARG conjugative transfer between suspended bacteria and microplastic biofilms. The results demonstrated that, compared to suspended bacteria, microplastic biofilms enhanced the frequency of ARG conjugative transfer by 7.2–19.6 times. Among them, biofilms on polyethylene microplastics showed the strongest promotion of conjugation. After the formation of microplastic biofilms, there was a significant increase in bacterial density within the biofilms, which raised the collision frequency of donor and recipient bacteria. Then microplastic biofilms facilitated the gene expression levels of outer membrane proteins, enhanced bacterial gene transfer capabilities, promoted the synthesis of conjugative pili, accelerated the formation of conjugative pairing systems, and elevated the expression levels of genes related to DNA replication and transfer systems, thereby enhancing the conjugative transfer of ARGs within microplastic biofilms. Among different types of microplastic biofilms, polyethylene biofilms exhibited the highest bacterial density, thus showing the highest frequency of ARG conjugation. This study highlights the risks associated with ARG conjugative transfer following the formation of microplastic biofilms and provides insights into the risks of microplastic and antibiotic resistance propagation in estuarine environments.

Comparative Study of Suspended and Immobilized Bacteria Isolated from Garage Soil on Vegetable Oil Degradation

As an essential part of the ecosystem, soil helps to support life by promoting interactions between bedrock, water, air, and biota. But the growing number of oil spills has raised serious environmental issues, presenting problems similar to petroleum hydrocarbon pollution. Vegetable oils are generally thought to be safe, but because of their toxicity and persistence, they may have negative environmental impacts. Bioremediation techniques have gained popularity as a sustainable method of cleaning up hydrocarbon-contaminated locations in response to this environmental problem. This study uses immobilized microbial technology to increase the bioremediation effectiveness by examining the ability of bacteria isolated from a garage oil spill region to degrade vegetable oil. The study compares the degradation rates with naturally occurring bacteria to assess the effectiveness of the detected bacterium’s breakdown over time in both suspended and immobilized situations. The results add to our knowledge of microbial-mediated bioremediation in contaminated environments and highlight the promise of immobilized microbial technology as a workable method for reducing vegetable oil spills. After 45 days, immobilized bacteria showed a drastic reduction in hydrocarbon content compared to suspended bacteria, indicating increased efficacy. Moreover, the degradation rate of immobilized bacteria was significantly higher than that of suspended bacteria, as observed from the difference in hydrocarbon concentration between day 1 and day 45. Statistical analysis confirmed these findings, highlighting the effectiveness of immobilized bacteria in hydrocarbon degradation.

Crowding accelerates the rotation of a bacterial rotor

Understanding the propulsion of a swimmer in a large group of individuals holds the key to unravelling the intriguing dynamics of active matter collective motion. Here, we develop a two-dimensional (2-D) self-assembled rotor, powered by bacterial flagella. At a water–air interface, the average direction of rotation of a rotor is fixed. When the chiral rotor is put into a 2-D bacterial suspension, we examine the average and fluctuation of the angular velocity of the rotor. Remarkably, the average angular velocity of a rotor is found to increase up to 3 times when the density of surrounding bacterial suspension increases and the increase is nonlinear. In a dense suspension of bacteria, the existence of a rotor disrupts vortices in the surrounding active turbulence, and the acceleration of the rotor is independent of the activity level of the surrounding free bacteria. The nonlinear acceleration thus results from hydrodynamic interaction with surrounding crowdedness that can be quantitatively explained by hydrodynamic simulation. The simultaneity between the acceleration of rotor and free bacteria in active turbulence suggests that crowding-induced acceleration may promote the onset of instability. The result will inspire new active-matter-based microfluidic devices with improved transport properties.

Enhancing biocementation performance in low permeability clayey soil through sand column strategy

Soil erosion is a common phenomenon which causes lots of geological and engineering disasters. Clayey soil erosion control is a hot research topic and a challenging issue for its low permeability and lack of effective infiltration of treatment solutions. In this study, a coupling microbially induced calcium carbonate precipitation (MICP)-sand column method was proposed as a promising and sustainable technique to mitigate surface clayey soil erosion with adjustable treatment depth. Seven groups of soil samples were prepared, including pure soil, MICP-treated soil, and MICP-sand column method-treated soil. A series of disintegration tests and penetration tests were conducted to investigate the method feasibility and adjusting mechanism of erosion mitigation with varying sand column heights and diameters. Compared to pure soil sample, sample treated by MICP-sand column (6 cm-height and 3 cm-diameter) method could reach a maximum reduction of 50 % in ultimate disintegration rate and a 30 % increase in the highest penetration resistance. The sample has a 7.9 cm effective treatment depth, which is 5.3 times of the MICP-treated sample. The mechanism of erosion mitigation can be attributed to that the sand column serves as a favorable path for the bacteria suspension and cementation solution in low-permeability-clayey soil and improves the effective depth of MICP treatment. Adjusting the height and diameter of sand column can change the calcium carbonate distribution, especially at the locations near the surface and around the sand column. In this study, the optimal height and diameter are 6 cm and 3 cm, respectively, and finally form a U-shape three-layer structure. The structure significantly increases the proportion of the hard crust layer and weak-cemented layer with high hydro-mechanical properties. In conclusion, the coupling MICP-sand column method with reasonable height and diameter of sand column shows the ability to control surface erosion of soil and has better long-term mitigation performance.

Carbon metabolism of a novel isolate from Lacticaseibacillus rhamnosus Probio-M9 derived through space mutant.

Carbon source is a necessary nutrient for bacterial strain growth. In industrial production, the cost of using different carbon sources varies greatly. Moreover, the complex environment in space may cause metabolic a series of changes in the strain, and this method has been successfully applied in some basic research. To date, space mutagenesis is still limited number of studies, particularly in carbon metabolism of probiotics. HG-R7970-41 was isolated from bacterium suspension (Probio-M9) after space flight, which can produce capsular polysaccharide after space mutagenesis. Phenotype Microarray (PM) was used to evaluated the metabolism of HG-R7970-41 in 190 single carbon sources. RNA sequencing and total protein identification of two strains revealed their different carbon metabolism mechanisms. PM results demonstrated the metabolism of 10 carbon sources were different between Probio-M9 and HG-R7970-41. Transcriptomic and proteomic analyses revealed that this change in carbon metabolism of HG-R7970-41 mainly related to changes in phosphorylation and the glycolysis pathway. Based on the metabolic mechanism of different carbon sources and related gene cluster analysis, we found that the final metabolic activities of HG-R7970-41 and Probio-M9 were mainly regulated by PTS-specific membrane embedded permease, carbohydrate kinase and two rate-limiting enzymes (phosphofructokinase and pyruvate kinase) in the glycolysis pathway. The expanded culture test also confirmed that HG-R7970-41 had different metabolic characteristics from original strain. These results suggested that space environment could change carbon metabolism of Probio-M9. The new isolate (HG-R7970-41) showed a different carbon metabolism pattern from the original strain mainly by the regulation of two rate-limiting enzymes.

The dual role of biochar in boosting phenol biodegradation and anti-inhibition performance against hazardous substrate

Enhancing the biodegradation of hazardous recalcitrant organic compounds from wastewater has garnered increasing attention. Herein, bamboo biochar was screened for subsequent investigation as it considerably enhanced hazardous phenol removal efficiency by 125.68 % over the biochar-free system at 700 mg/L. Besides, the influencing factors were optimized, including biochar dosage and phenol concentration. However, the phenol removal rate decreased by 75.91 % within 15 hours when utilizing biochar oxidized by H2O2, perhaps due to eliminating aromatic clusters on biochar. Combined with FTIR analysis, CO and -OH on aromatic clusters of biochar were recognized as the underlying contributors to facilitating electron transfer in phenol biodegradation. Furthermore, the ortho-disubstituted benzene ring was discovered on biochar, which may originate from catechol, an intermediate during phenol degradation, suggesting that biochar also serves as a site for phenol biodegradation. Moreover, the fabricated biochar-sodium alginate microspheres enhanced anti-inhibition to phenol over suspended bacteria by 192.15 %. The system also eliminated 99.93 % of phenol after the seven recycles at 1258.75 mg/L, exhibiting excellent reusability. This work provides valuable insights into bioaugmentation strategies for bio-degrading phenol using biochar.

Effects of Nano-Titanium Dioxide on the Horizontal Transfer of Antibiotic Resistance Genes in Microplastic Biofilms

Emerging pollutants such as microplastics in water environments readily accumulate microorganisms on their surfaces, forming biofilms and concentrating antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). Consequently, microplastic biofilms have attracted the attention of researchers. Horizontal gene transfer (HGT) of ARGs is one of the primary ways that bacteria acquire antibiotic resistance. Most studies focus on the effects of nanomaterials on suspended bacteria, but microplastic biofilms as hotspots for horizontal gene transfer also warrant significant investigation. This study primarily explored and compared the effects of nano-titanium dioxide on the conjugation transfer frequency of ARGs in suspended bacteria and microplastic biofilms. Nano-titanium dioxide could promote ARG conjugation in both suspended bacteria and microplastic biofilms, with a greater effect on the former. The mechanism involved nano-titanium dioxide promoting the production of reactive oxygen species (ROS) in suspended and biofilm bacteria, increasing the synthesis of outer membrane proteins, enhancing the cell membrane permeability, and elevating the expression levels of conjugation-related genes, thereby facilitating the conjugation transfer of ARGs. Biofilm bacteria, being heavily encased and protected by extracellular polymeric substances (EPS), exhibit greater resistance to external environmental pressure, resulting in the weaker impact of nano-titanium dioxide on biofilm bacteria compared to suspended bacteria. This study reveals the risk of ARG conjugation transfer within microplastic biofilms induced by nanomaterials, providing valuable insights into the risks of microplastic and antibiotic resistance dissemination in water environments.

A New Guanidine-Core Small-Molecule Compound as a Potential Antimicrobial Agent against Resistant Bacterial Strains.

The guanidine core has been one of the most studied functional groups in medicinal chemistry, and guanylation reactions are powerful tools for synthesizing this kind of compound. In this study, a series of five guanidine-core small molecules were obtained through guanylation reactions. These compounds were then evaluated against three different strains of Escherichia coli, one collection strain from the American Type Culture Collection (ATCC) of E. coli ATCC 35218, and two clinical extended-spectrum beta-lactamase (ESBL)-producing E. coli isolates (ESBL1 and ESBL2). Moreover, three different strains of Pseudomonas aeruginosa were studied, one collection strain of P. aeruginosa ATCC 27853, and two clinical multidrug-resistant isolates (PA24 and PA35). Among Gram-positive strains, three different strains of Staphylococcus aureus, one collection strain of S. aureus ATCC 29213, and two clinical methicillin-resistant S. aureus (MRSA1 and MRSA2) were evaluated. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) experiments were reported, and the drop plate (DP) method was used to determine the number of viable suspended bacteria in a known beaker volume. The results from this assessment suggest that guanidine-core small molecules hold promise as therapeutic alternatives for treating infections caused by clinical Gram-negative and Gram-positive bacteria, highlighting the need for further studies to explore their potential. The results from this assessment suggest that the chemical structure of CAPP4 might serve as the basis for designing more active guanidine-based antimicrobial compounds, highlighting the need for further studies to explore their potential.

Thinning by cluster breaking: Active matter and shear flows share thinning mechanisms

Among many unexpected phenomena of active matter is the recently observed superfluid-like thinning (viscosity drop) behavior of bacteria suspensions. Understanding this peculiar self-propelled thinning by active matter is of theoretical and practical importance. Here, we find that, although distinct in driving mechanisms, active matter and shear flows exhibit similar thinning behaviors upon the increase of self-propulsion and shear forces, respectively. Our structural characterizations reveal that they actually share the same cluster-breaking mechanism of thinning. How fast and how shattered the cluster is broken determines the (dis)continuity of the thinning. This explains why adding active particles to Newtonian fluids can cause thinning, in which rotation of active particles play a key role in breaking clusters. Our work proposes a mechanism of self-propelled thinning and further establishes the underlying connections between active matter and shear flows.

In vitro Antibacterial Effect Study of Plasma-Activated Saline on Mycobacterium Tuberculosis.

This study aimed to investigate the antibacterial effects of plasma-activated saline (PAS) on My-cobacterium tuberculosis (Mtb). We conducted a growth assay on 3 strains of Mtb and an antibiotic sensitivity test on 4 strains of Mtb. Both tests included groups treated with normal saline (NS), PAS, and hydrochloric acid (HCl). The test of antibiotic sensitivity consisted of parallel tests with two concentrations of bacteria suspension: 10-2 and 10-4. The selected antibiotics were rifampicin (RIF), isoniazid (INH), ethambutol (EMB), and streptomycin (SM). The number of bacteria was determined after one month of culture under different conditions. The Kruskal-Wallis test was used to analyze the differences in grouping factors at representative time points. The growth assay indicated that PAS significantly inhibited the growth of 3 strains of Mtb compared with NS and HCl treatment groups. Furthermore, except for the initial observation time point, the remaining three observation time points consistently demonstrate no significant differences between the NS group and the HCl group. The antibiotic sensitivity test of INH, SM, and RIF indicated that PAS could inhibit the growth of antibiotic-resistant Mtb, and the antibiotic sensitivity test of INH and SM with bacterial suspension concentration of 10-2 and SM with bacterial suspension concentration of 10-4 showed statistically different results. The antibiotic sensitivity test of EMB indicated that the growth of Mtb in PAS was slower than that in NS and HCl in both antibiotic-resistant and sensitive Mtb, but there was no statistical difference. The study indicates that PAS contains a significant amount of active substances and exhibits high oxidizability and an acidic pH state. The unique physicochemical properties of PAS significantly delayed the growth of Mtb, compared to the NS and the HCl. PAS not only inhibited the growth of drug-sensitive strains but also significantly enhanced the sensitivity of drug-resistant strains to anti-tuberculosis drugs, which may provide a new therapeutic strategy for the treatment of tuberculosis.

Enhanced antimicrobial efficacy and energy efficiency of low irradiance 405-nm light for bacterial decontamination

Due to its increased safety over ultraviolet light, there is interest in the development of antimicrobial violet-blue light technologies for infection control applications. To ensure compatibility with exposed materials and tissue, the light irradiances and dose regimes used must be suitable for the target application. This study investigates the antimicrobial dose responses and germicidal efficiency of 405 nm violet-blue light when applied at a range of irradiance levels, for inactivation of surface-seeded and suspended bacteria. Bacteria were seeded onto agar surfaces (101–108 CFUplate−1) or suspended in PBS (103–109 CFUmL−1) and exposed to increasing doses of 405-nm light (≤ 288 Jcm−2) using various irradiances (0.5–150 mWcm−2), with susceptibility at equivalent light doses compared. Bacterial reductions ≥ 96% were demonstrated in all cases for lower irradiance (≤ 5 mWcm−2) exposures. Comparisons indicated, on a per unit dose basis, that significantly lower doses were required for significant reductions of all species when exposed at lower irradiances: 3–30 Jcm−2/0.5 mWcm−2 compared to 9–75 Jcm−2/50 mWcm−2 for low cell density (102 CFUplate−1) surface exposures and 22.5 Jcm−2/5 mWcm−2 compared to 67.5 Jcm−2/150 mWcm−2 for low density (103 CFUmL−1) liquid exposures (P ≤ 0.05). Similar patterns were observed at higher densities, excluding S. aureus exposed at 109 CFUmL−1, suggesting bacterial density at predictable levels has minimal influence on decontamination efficacy. This study provides fundamental evidence of the greater energy efficacy of 405-nm light for inactivation of clinically-significant pathogens when lower irradiances are employed, further supporting its relevance for practical decontamination applications.

The Effect of Bacillus Pumilus on the Free Swell and Atterberg Limits of Black Cotton Soil for use in Road Subgrades

The study evaluated the potential utilisation of Bacillus pumilus (B. pumilus) induced calcite precipitate for the improvement of the engineering properties of black cotton soil in road construction which is an eco-friendly technique and forms part of what is referred to as green engineering. The effect of B. pumilus on the free swell and Atterberg limits of black cotton soil for use in road subgrades was studied using microbial induced calcite precipitation (MICP) method. Five stepped B. pumilus suspension density of 0, 1.5 x 108, 6.0 x 108, 1.2 x 109, 1.8 x 109 and 2.4 x 109 cells/ml, respectively, were utilized for the treated specimens, while only cementitious reagent acted as the control specimen. Three (3) mix ratios of % Bacteria suspension (B) - %Cementitious reagent (C) (i.e., 25B - 75C, 50B - 50C and 75B - 25C) were adopted. It was observed that free swell treated with 25B - 75C decreased from 70 % and 50 % for the natural and control specimens, respectively, to minimum value of 46 %, with increasing B. pumilus suspension density at 2.4 × 109 cells/ml. Atterberg limits results showed liquid limit values decreased from 53 % and 62 % for the natural and control samples, respectively, to 43 % for samples treated with 25B - 75C at B. pumilus suspension density of 2.4 x 109 cells/ml, while the plastic limit increased from 28.2 % for the control soil sample to 30 % for the 75B - 25C mix ratio at B. pumilus suspension density of 2.4 x 109 cells/ml. Tests results indicated a considerable improvement with 25B - 75C mix ratio at B. pumilus suspension density of 2.4 × 109 cells/ml which significantly reduced the free swell, while the Atterberg limits fell below Nigerian General Specifications requirements of not greater than 35% Liquid limit, 12% Plasticity index and 35% passing No.200 sieve for utilization as road subgrades. However, it could be used as a subgrade material in low-volume roads.

Lethal puncturing of planktonic Gram-positive and Gram-negative bacteria by magnetically-rotated silica hexapods

Planktonic bacterial presence in many industrial and environmental applications and personal health-care products is generally countered using antimicrobials. However, antimicrobial chemicals present an environmental threat, while emerging resistance reduces their efficacy. Suspended bacteria have no defense against mechanical attack. Therefore, we synthesized silica hexapods on an α-Fe2O3 core that can be magnetically-rotated to inflict lethal cell-wall-damage to planktonic Gram-negative and Gram-positive bacteria. Hexapods possessed 600 nm long nano-spikes, composed of SiO2, as shown by FTIR and XPS. Fluorescence staining revealed cell wall damage caused by rotating hexapods. This damage was accompanied by DNA/protein release and bacterial death that increased with increasing rotational frequency up to 500 rpm. Lethal puncturing was more extensive on Gram-negative bacteria than on Gram-positive bacteria, which have a thicker peptidoglycan layer with a higher Young's modulus. Simulations confirmed that cell-wall-puncturing occurs at lower nano-spike penetration levels in the cell walls of Gram-negative bacteria. This approach offers a new way to kill bacteria in suspension, not based on antimicrobial chemicals.

Utilizing fab fragment-conjugated surface plasmon resonance-based biosensor for detection of Salmonella Enteritidis.

Although antibodies, a key element of biorecognition, are frequently used as biosensor probes, the use of these large molecules can lead to adverse effects. Fab fragments can be reduced to allow proper antigen-binding orientation via thiol groups containing Fab sites that can directly penetrate Au sites chemically. In this study, the ability of the surface plasmon resonance (SPR) sensor to detect Salmonella was studied. Tris(2-carboxyethyl)phosphine was used as a reducing agent to obtain half antibody fragments. Sensor surface was immobilized with antibody, and bacteria suspensions were injected from low to high concentrations. Response units were changed by binding first reduced antibody fragments, then bacteria. The biosensor was able to determine the bacterial concentrations between 103 and 108 CFU/mL. Based on these results, the half antibody fragmentation method can be generalized for faster, label-free, sensitive, and selective detection of other bacteria species.

A novel rapid bioluminescence-based antimicrobial susceptibility testing method based on adenosine triphosphate consumption.

Standard, phenotypic antimicrobial susceptibility testing (AST) methods require 16-20 h of incubation and are considered as the bottleneck in providing timely input for appropriate antimicrobial treatment. In this study, a novel adenosine triphosphate (ATP)-bioluminescence-based method which allows rapid AST within 3 h was described. Standard AST was performed for 56 Enterobacterales isolates using EUCAST disk diffusion (DD) methodology. For the bioluminescence-based rapid AST, suspensions of bacteria were prepared using Mueller-Hinton broth to obtain a turbidity of 0.5 McFarland. The suspensions were distributed into 96-well microtiter plates. ATP (20 mM) and fixed concentrations of different antibiotics were added. Following incubation at 37°C for 1 h, a luminescent reaction mixture, including the substrate luciferin and luciferase enzyme solutions, was added. The chemiluminescence was monitored using an imaging system. Light production demonstrated the presence of ATP, indicating that the isolate was susceptible to the antibiotic in the well. Absence or decrease of light intensity, compared with the growth control well, indicated the use of ATP as an indirect measure of bacterial growth, and therefore resistance to the antibiotic in the well. The novel AST method was tested using a total of 348 test wells. Concordance was achieved for 290 (83.3%) of the tests, whereas 52 (14.9%) and 6 (1.7%) tests caused minor and major errors, respectively. In this study, a bioluminescence-based rapid AST was developed based on the consumption of ATP by bacteria. Our method's uniqueness relies on determining ATP consumption by microorganisms in the presence or absence of an antibiotic. The novel AST method described in this study lays the groundwork for obtaining rapid results, which should be considered as a proof of concept. With further optimization studies, this novel method can provide higher accuracy and be introduced into clinical practice as a routine AST method.

Induced Partial Saturation Using Pseudomonas stutzeri Biogas for Mitigate Structure Settlement

Induced partial saturation (IPS) is a new foundation treatment method for mitigating soil liquefaction using biogas. A series of laboratory tests were performed to demonstrate the influencing factors of IPS using Pseudomonas stutzeri biogas. On the basis of the optimal biogas production conditions, the intervention effect of Pseudomonas stutzeri biogas on the foundation deformation under buildings was investigated based on shaking table tests. The test results showed that the best carbon source in the denitrification process of Pseudomonas stutzeri biogas is sodium citrate. The most effective initial value of optical density-based concentration was 0.1. The carbon–nitrogen ratio (C/N) of the bacterium suspension was used as the index to control the saturation. The degree of saturation reduction showed a good linear correlation with the C/N. The optimum temperature of this method was between 20 °C and 30 °C. The most suitable pH value was between 7 and 9. The environmental factors had minimal influence on the degree of saturation reduction but had a significant effect on the average rate of gas generation and the period of initial stagnation. After Pseudomonas stutzeri biogas desaturation, the settlement of the building was greatly reduced. The settlement of saturation of 92.5% sand foundation reached 17.1 mm, and the 85% saturation was only 10.6 mm. These results provide a good foundation for the feasibility of utilizing Pseudomonas stutzeri biogas mitigation of the liquefaction hazard of sand.

Microbiome Structure of Activated Sludge after Adaptation to Landfill Leachate Treatment in a Lab-Scale Sequencing Batch Reactor

During adaptation to waters that are rich in xenobiotics, biological systems pass through multiple stages. The first one is related to the restructuring of communities, pronounced destruction of the structure, and multiplication of active biodegradants. The purpose of the present research was to describe the microbiome restructuring that occurs during the adaptation stage in landfill leachate treatment. In a model SBR (sequencing batch reactor), a 21-day purification process of landfill leachate was simulated. Wastewater was fed in increasing concentrations. When undiluted leachate entered, the activated sludge structure disintegrated (Sludge Volume Index—4.6 mL/g). The Chemical Oxygen Demand and ammonium nitrogen concentration remained at high values in the influent (2321.11 mgO2/L and 573.20 mg/L, respectively). A significant amount of free-swimming cells was found, and the number of aerobic heterotrophs and bacteria of the genera Pseudomonas and Acinetobacter increased by up to 125 times. The Azoarcus-Thauera cluster (27%) and Pseudomonas spp. (16%) were registered as the main bacterial groups in the activated sludge. In the changed structure of the microbial community, Gammaproteobacteria, family Rhizobiaceae, class Saccharimonadia were predominantly represented. Among the suspended bacteria, Microbactericeae and Burkholderiaceae, which are known for their ability to degrade xenobiotics, were present in larger quantities. The enzymological analysis demonstrated that the ortho-pathway of cleavage of aromatic structures was active in the community. The described changes in the leachate-purifying microbial community appear to be destructive at the technological level. At the microbiological level, however, trends of initial adaptation were clearly outlined, which, if continued, could provide a highly efficient biodegradation community.