Pseudomonas Aeruginosa Strains Research Articles

Preventive nasal administration of flagellin restores antimicrobial effect of gentamicin and protects against a multidrug-resistant strain of Pseudomonas aeruginosa.

The increasing prevalence of multidrug-resistant Pseudomonas aeruginosa (PA) is a significant concern for chronic respiratory disease exacerbations. Host-directed drugs, such as flagellin, an agonist of toll-like receptor 5 (TLR5), have emerged as a promising solution. In this study, we evaluated the prophylactic intranasal administration of flagellin against a multidrug-resistant strain of PA (PAMDR) in mice and assessed the possible synergy with the antibiotic gentamicin (GNT). The results indicated that flagellin treatment before infection decreased bacterial load in the lungs, likely due to an increase in neutrophil recruitment, and reduced signs of inflammation, including proinflammatory cytokines. The combination of flagellin and GNT showed a synergistic effect, decreasing even more the bacterial load and increasing mice survival rates, in comparison to mice pre-treated only with flagellin. These findings suggest that preventive nasal administration of flagellin could restore the effect of GNT against MDR strains of PA, paving the way for the use of flagellin in vulnerable patients with chronic respiratory diseases.

Comparative genomic analysis of Pseudomonas aeruginosa strains susceptible and resistant to carbapenems and aztreonam isolated from patients with healthcare-associated infections in a Mexican hospital.

Pseudomonas aeruginosa (PA) is an important opportunistic pathogen that causes different infections on immunocompromised patients. Within PA accessory genome, differences in virulence, antibiotic resistance and biofilm formation have been described between strains, leading to the emergence of multidrug-resistant strains. The genome sequences of 17 strains isolated from patients with healthcare-associated infections in a Mexican hospital were genomically and phylogenetically analyzed and antibiotic resistance genes, virulence genes, and biofilm formation genes were detected. Fifteen of the 17 strains were resistant to at least two of the carbapenems meropenem, imipenem, and the monobactam aztreonam. The antibiotic resistance (mexA, mexB, and oprM) and the biofilm formation (pslA and pslD) genes were detected in all strains. Differences were found between strains in accessory genome size. The strains had different sequence types, and seven strains had sequence types associated with global high risk epidemic PA clones. All strains were represented in two groups among PA global strains. In the 17 strains, horizontally acquired resistance genes to aminoglycosides and beta-lactams were found, mainly, and between 230 and 240 genes that encode virulence factors. The strains under study were variable in terms of their accessory genome, antibiotic resistance, and virulence genes. With these characteristics, we provide information about the genomic diversity of clinically relevant PA strains.

Characterization of the Outer Membrane Vesicles of Pseudomonas aeruginosa Exhibiting Growth Inhibition against Acinetobacter baumannii.

We investigated the Pseudomonas aeruginosa (PA) outer membrane vesicles (OMVs) and their effect on Acinetobacter baumannii (AB) growth in vitro. The inhibitory effects of PA on AB were assessed using a cross-streak assay. The OMVs were extracted through high-speed centrifugation, tangential flow filtration, and ultracentrifugation and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transmission electron microscopy (TEM), and nanoparticle tracking assays (NTAs). Proteomic analysis was conducted to compare the OMVs of different PA strains. PA022 exhibited more pronounced inhibition of AB growth compared with PA ATCC 27853. TEM confirmed the presence of OMVs in both PA022 and PA ATCC 27853, revealing phospholipid bilayer structures. The NTA revealed similar sizes and concentrations. Proteomic analysis identified 623 and 538 proteins in PA022 and PA ATCC 27853 OMVs, respectively, with significant proportions of the outer membrane and extracellular proteins, respectively. Importantly, PA022 OMVs contained six known virulence factors and motility-associated proteins. This study revealed the unique characteristics of PA OMVs and their inhibitory effects on AB growth, shedding light on their role in bacterial interactions. Proteomic analysis provides valuable insights into potential pathogenic functions and therapeutic applications against bacterial infections.

Pseudomonas Aeruginosa Affects the Function of Pulmonary Vascular Endothelial Cells

Objective To investigate the impact of Pseudomonas aeruginosa(PA) infection on the function of pulmonary vascular endothelial cells,and explore the mechanism of this bacterium in exacerbating lung inflammation in mice. Methods Two hours after human lung microvascular endothelial cell(HULEC-5a) were infected with the PA strain PAO1,the mRNA levels of autophagy-related gene 5(ATG5),6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3(PFKFB3),and calcium adhesion protein 5(CDH5) were determined by reverse transcription real-time fluorescent quantitative PCR(RT-qPCR).The protein levels of ATG5,PFKFB3,and vascular endothelial calcium adhesion protein(VE-cadherin) were detected by immunofluorescence.After the expression of ATG5 and PFKFB3 was respectively knocked down by small interfering RNA(siRNA),RT-qPCR was employed to measure the mRNA levels of ATG5,PFKFB3,and CDH5,and immunofluorescence to detect the protein levels of PFKFB3 and VE-cadherin.In addition,the lactate assay kit was used to determine the level of lactate in the cells.After mice were infected with PAO1,lung inflammation was assessed through histopathological section staining.Confocal microscopy was employed to capture and analyze fluorescence-labeled PFKFB3 and VE-cadherin in endothelial cells. Results Compared with the control group,the HULEC-5a cells infected with PAO1 showed up-regulated mRNA and protein levels of PFKFB3(all P<0.05),down-regulated mRNA level of CDH5(P=0.023),disrupted continuity and down-regulated protein level of VE-cadherin(P<0.001),and elevated lactate level(P=0.017).Compared with PAO1-infected HULEC-5a cells,knocking down PFKFB3 led to the up-regulated mRNA level of CDH5(P=0.043),lowered lactate level(P=0.047),and restored continuity of VE-cadherin;knocking down ATG5 led to up-regulated mRNA and protein levels of PFKFB3(P=0.013 and P=0.003),elevated lactate level(P=0.015),and down-regulated mRNA level of CDH5(P=0.020) and protein level of VE-cadherin(P=0.001).The HE staining results showed obvious red blood cell leakage,inflammatory cell infiltration,alveolar septal widening,and partial detachment of vascular endothelial cells in the alveoli of PA-infected mice.Immunofluorescence staining showed up-regulated expression of PFKFB3 and decreased fluorescence signal of VE-cadherin in endothelial cells of infected mice compared with normal mice. Conclusion PA may regulate the PFKFB3 pathway via AGT5 to disrupt the function of pulmonary vascular endothelial cells,thereby exacerbating the inflammation in the lungs of mice.

Therapeutic strategies for drug-resistant Pseudomonas aeruginosa: Metal and metal oxide nanoparticles.

Pseudomonas aeruginosa (PA) is a widely prevalent opportunistic pathogen. Multiple resistant strains of PA have emerged from excessive or inappropriate use of antibiotics, making their eradication increasingly difficult. Therefore, the search for highly efficient and secure novel antimicrobial agents is crucial. According to reports, there is an increasing exploration of nanometals for antibacterial purposes. The antibacterial mechanisms involving the nanomaterials themselves, the release of ions, and the induced oxidative stress causing leakage and damage to biomolecules are widely accepted. Additionally, the study of the cytotoxicity of metal nanoparticles is crucial for their antibacterial applications. This article summarizes the types of metal nanomaterials and metal oxide nanomaterials that can be used against PA, their respective unique antibacterial mechanisms, cytotoxicity, and efforts made to improve antibacterial performance and reduce toxicity, including combination therapy with other materials and antibiotics, as well as green synthesis approaches.

Polyclonal anti-whole cell IgY passive immunotherapy shields against P. aeruginosa-induced acute pneumonia and burn wound infections in murine models

Pseudomonas aeruginosa (PA) is a multidrug-resistant (MDR) opportunistic pathogen causing severe hospital-, and community-acquired infections worldwide. Thus, the development of effective immunotherapy-based treatments is essential to combat the MDR-PA infections. In the current study, we evaluated the protective efficacy of polyclonal avian antibodies raised against inactivated whole cells of the PAO1 strain in murine models of acute pneumonia and burn wound. The efficacy of generated antibodies was evaluated against different PA strains through several in vitro, ex vivo and in vivo experiments. The results showed that the anti-PAO1-IgY effectively reduced the motility, biofilm formation and cell internalization ability, and enhanced the opsonophagocytic killing of PA strains through the formation of immobilized bacteria and induction of increased cell surface hydrophobicity. Furthermore, immunotherapy with anti-PAO1-IgY completely protected mice against all PA strains in both acute pneumonia and burn wound murine models. It was found to reduce the bacterial loads in infected burned mice through interfering with virulence factors that play vital roles in the early stages of PA infection, such as colonization and cell internalization. The immunotherapy with anti-PAO1-IgYs could be instrumental in developing effective therapies aimed at reducing the morbidity and mortality associated with PA infections.

The Impact of Pseudomonas aeruginosa Infection in Adult Cystic Fibrosis Patients-A Single Polish Centre Study.

Pseudomonas aeruginosa (PA) is one of the most predominant pathogens of lung infections, often causing exacerbations in adult patients with cystic fibrosis (CF). Microbiological characterization of 74 PA isolates and to evaluate the correlations between the bacterial features and 44 adult Polish CF cohort clinical parameters. The most common variant in the CF transmembrane conductance regulator (CFTR) gene was F508del (76.3%), followed by 3849+10kbC>T (26.3%). A total of 39.4% of the PA isolates showed multiple resistances. In patients with parameters pointing to a decline in lung function, there was a statistically significant moderate correlation with β-lactam resistance and a weak correlation between hospital frequency and colistin resistance. The mucoidity did not correlate with the biofilm formation ability, which showed 41.9% of the isolates. Proteolytic activity, observed in 60.8% of the clinical isolates, was weakly associated with motility detected in 78.4% of the strains. The genetic profiles of the PA were highly heterogeneous, and a weak positive correlation was established between cluster group and biofilm formation. The findings suggest that there is a high variety in P. aeruginosa populations in adult CF patients. There is a need to monitor PA strains in groups of patients with cystic fibrosis, in particular, in terms of the occurrence of antibiotic resistance related to a decline in lung function.

Экспериментальное лечение эндофтальмита, вызванного синегнойной инфекцией с множественной лекарственной устойчивостью, с помощью конъюгатов на основе наночастиц

Relevance. Pseudomonas aeruginosa (PA) is an extremely dangerous microorganism whose elimination from the host organism is very difficult due to the presence of multidrug resistance (MDR) factors in PA. The outcome of endophthalmitis (EP) mediated by PA in 50% of cases will be evisceration of the eyeball, and more than 95% of patients receive visual acuity – hand movements near the face and below. In this regard, studies of quantum dots (QDs), which have shown high anti-infective activity in laboratory conditions, are of particular relevance. Purpose. Evaluation of the effectiveness of the treatment of EP caused by PA with MDR using QDs-based conjugates in the experiment. Material and methods. The object of the study was laboratory rabbits of the Chinchilla breed (n=27), male, aged from 3 to 3.5 months. As an EP inducer, all laboratory animals underwent intravitreal injection (IVI) of a multidrug-resistant PA strain in a volume of 0.05 ml in the right eye. QDs of the Ag (10%):InP/ZnS MPA type were taken as the basis for the creation of conjugates with antibiotics (AB). After the manifestation of EP, the animals underwent intravitreal injections (IVI) of dosage forms in accordance with the distribution into the following subgroups, 3 rabbits in each. Subgroups No. 1–3 – IVI 2.25 mg/0.1 ml Ceftazidime. Subgroups No. 4–6 – IVI 0.1 ml of QDs solution at concentrations of 10%, 0.1%, 0.01%, respectively. Subgroups No. 7–9 – IVI solution of QDs 0.05 ml (at concentrations of 10%, 0.1%, 0.01%, respectively) in combination with 2.25 mg/0.05 ml of Ceftazidime (conjugates). Efficacy was evaluated based on clinical parameters: the dynamics of hypopyon resorption, the presence of inflammatory cells in the anterior chamber, the dynamics of regression of pathological suspension in the vitreal cavity, the dynamics of pathological neovascularization of the iris, the dynamics of the cornea. Results. Rabbits from subgroups No. 1–3 (1st group) were characterized by the least dynamics in relation to the regression of symptoms during the 7-day observation period. Rabbits from subgroups No. 4–6 (2nd group) were characterized by moderate positive dynamics in relation to the regression of symptoms. Rabbits from subgroups No. 7–9 (3rd group) were characterized by the best positive dynamics in relation to the regression of symptoms. Conclusion. It was possible to prove the effectiveness of the treatment of experimental antibiotic-resistant EP caused by a strain of PA with MDR by means of QDs Ag(10%):InP/ZnS MPA at concentrations of 10%, 0.1% and 0.01%, especially in combination with 2.25 mg/0.05 ml of Ceftazidime in as a conjugate. Based on the results obtained, it can be assumed that active research and the use of QDs as conjugates with modern ABs can solve the overall problem of combating MDR strains. Key words: Pseudomonas Aeruginosa, quantum dots, endophthalmitis, multidrug resistance, antibiotic resistance, conjugates

Development of Quinazolinone Derivatives as Modulators of Virulence Factors of Pseudomonas aeruginosa Cystic Fibrosis Strains.

Pseudomonas aeruginosa (PA), one of the ESKAPE pathogens, is an opportunistic Gram-negative bacterium responsible for nosocomial infections in humans but also for infections in patients affected by AIDS, cancer, or cystic fibrosis (CF). Treatment of PA infections in CF patients is a global healthcare problem due to the ability of PA to gain antibiotic tolerance through biofilm formation. Anti-virulence compounds represent a promising approach as adjuvant therapy, which could reduce or eliminate the pathogenicity of PA without impacting its growth. Pyocyanin is one of the virulence factors whose production is modulated by the Pseudomonas quinolone signal (PQS) through its receptor PqsR. Different PqsR modulators have been synthesized over the years, highlighting this new powerful therapeutic strategy. Based on the promising structure of quinazolin-4(3H)-one, we developed compounds 7a-d, 8a,b, 9, 10, and 11a-f able to reduce biofilm formation and the production of virulence factors (pyocyanin and pyoverdine) at 50 µM in two PA strains responsible for CF acute and chronic infections. The developed compounds did not reduce the cell viability of IB3-1 bronchial CF cells, and computational studies confirmed the potential ability of novel compounds to act as potential Pqs system modulators.

Antimicrobial Resistance in Pseudomonas aeruginosa before and during the COVID-19 Pandemic.

Pseudomonas aeruginosa (PA) is a major Gram-negative opportunistic pathogen causing several serious acute and chronic infections in the nosocomial and community settings. PA eradication has become increasingly difficult due to its remarkable ability to evade antibiotics. Therefore, epidemiological studies are needed to limit the infection and aim for the correct treatment. The present retrospective study focused on PA presence among samples collected at the San Giovanni di Dio and Ruggi D'Aragona University Hospital in Salerno, Italy; its resistance profile and relative variations over the eight years were analyzed. Bacterial identification and antibiotic susceptibility tests were performed by VITEK® 2. In the 2015-2019 and 2020-2022 timeframes, respectively, 1739 and 1307 isolates of PA were obtained from respiratory samples, wound swabs, urine cultures, cultural swabs, blood, liquor, catheter cultures, vaginal swabs, and others. During 2015-2019, PA strains exhibited low resistance against amikacin (17.2%), gentamicin (25.2%), and cefepime (28.3%); moderate resistance against ceftazidime (34.4%), imipenem (34.6%), and piperacillin/tazobactam (37.7%); and high resistance against ciprofloxacin (42.4%) and levofloxacin (50.6%). Conversely, during the 2020-2022 era, PA showed 11.7, 21.1, 26.9, 32.6, 33.1, 38.7, and 39.8% resistance to amikacin, tobramycin, cefepime, imipenem, ceftazidime, ciprofloxacin, and piperacillin/tazobactam, respectively. An overall resistance-decreasing trend was observed for imipenem and gentamicin during 2015-2019. Instead, a significant increase in resistance was recorded for cefepime, ceftazidime, and imipenem in the second set of years investigated. Monitoring sentinel germs represents a key factor in optimizing empirical therapy to minimize the spread of antimicrobial resistance.

Sniffing Out Pseudomonas Aeruginosa Wound Infections with Vitamin B12: A Simple and Selective Colorimetric Biosensor for Early Diagnosis

Abstract Background The vacuum-assisted closure (VAC) system is a useful tool to promote wound healing. However, due to its occlusive system, the wound is often prone to Pseudomonas aeruginosa (PA) infections. Early detection and initiation of treatment is essential to limit the damage (e.g., saving a skin graft), but the black sponge occludes the wound and prevents its visual assessment leading to delayed diagnosis. Fortunately, each bacterium emits a spectrum of specific volatile organic compounds that can be used for detection. For PA, the signature compound is hydrogen cyanide. Aims We aimed to develop a colorimetric biosensor for “naked-eye” detection of PA that takes advantage of the negative pressure created by the VAC to collect hydrogen cyanide. Methods We used aquocyanocobinamide, a derivative of vitamin B12, as a colorimetric chemosensor of biological cyanide. By binding cyanide, the color of the sensor changes from orange to violet, which can be quantified with UV-VIS spectroscopy. The sensor was delivered onto the carrier material polyvinyl acetate. Its sensitivity was assessed using laboratory strains and clinical isolates of PA in-vitro and on an ex vivo skin model. Results UV-VIS spectroscopy readings of the sensor were calibrated for the cyanide concentration 0-250ppm. Cyanide produced by 30 laboratory and 20 clinical PA isolates were quantified based on the colorimetric changes of the sensor. While the sensor detected all PA strains, it did not falsely react to other wound pathogens such as Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli and Enterococci, suggesting a high specificity. In a proof-of-principle experiment, the colorimetric change of the sensor was detected by eye after 3 hours incubation with a PA culture. Conclusions The presence of Pseudomonas aeruginosa can be quickly and reliably detected by our vitamin B12-based colorimetric biosensor. Its incorporation inside the VAC-system can allow physicians to detect PA wound infections early on.

The efficacy of biosynthesized silver nanoparticles against Pseudomonas aeruginosa isolates from cystic fibrosis patients

The high antibiotic resistance of Pseudomonas aeruginosa (PA) makes it critical to develop alternative antimicrobial agents that are effective and affordable. One of the many applications of silver nanoparticles (Ag NPs) is their use as an antimicrobial agent against bacteria resistant to common antibiotics. The key purpose of this research was to assess the antibacterial and antibiofilm effectiveness of biosynthesized Ag NPs against six biofilm-forming clinically isolated strains of PA and one reference strain (ATCC 27853). Ag NPs were biosynthesized using a seed extract of Peganum harmala as a reducing agent. Ag NPs were characterized by Ultraviolet–visible (UV–Vis) spectroscopy and scanning transmission electron microscopy (STEM). The effect of Ag NPs on biofilm formation and eradication was examined through micro-titer plate assays, and the minimal inhibitory (MIC) and minimum bactericidal (MBC) concentrations determined. In addition, real-time polymerase chain reactions (RT-PCR) were performed to examine the effects of Ag NPs on the expression of seven PA biofilm-encoding genes (LasR, LasI, LssB, rhIR, rhII, pqsA and pqsR). The biosynthesized Ag NPs were spherically-shaped with a mean diameter of 11 nm. The MIC for each PA strain was 15.6 µg/ml, while the MBC was 31.25 µg/ml. All PA strains exposed to Ag NPs at sub-inhibitory concentrations (0.22–7.5 µg/ml) showed significant inhibitory effects on growth and biofilm formation. Biomass and biofilm metabolism were reduced dependent on Ag NP concentration. The expression of the quorum-sensing genes of all strains were significantly reduced at an Ag NP concentration of 7.5 µg/ml. The results demonstrate the extensive in-vitro antibacterial and antibiofilm performance of Ag NPs and their potential in the treatment of PA infection. It is recommended that future studies examine the possible synergy between Ag NPs and antibiotics.

Degree of Gelatination on Ag-Nanoparticles to Inactivate Multi-drug Resistant Bacterial Biofilm Isolated from Sewage Treatment Plant.

Overuse and improper dosage of antibiotics have generated antimicrobial resistance (AMR) worldwide. Pseudomonas aeruginosa (PA), a well-known bacterial strain can establish MDR leading to a variety of infections in humans. Furthermore, these PA strains hold the ability to form biofilms by generating extracellular polymeric substances on the surface of medical tools and critical care units. To supersede the infectious effect of MDR organisms, silver nanoparticles have been known to be the choice. Hence, the present study concentrates on the engineering of varying concentrations of gelatin-based polymeric hydrogel embedded with silver nanoparticles (G-AgNPs) for controlled bactericidal activity against MDR PA biofilms. Biofilms formation by MDR PA was confirmed microscopically and spectroscopy was taken as a tool to characterize and analyze the efficacy at every stage of experiments. When MDR PA biofilms were treated with G-AgNPs prepared with 5 % gelatin concentration (AgNP3), they exhibited superior bactericidal activity. Furthermore, a dose-dependent study showed that 800 nM of AgNP3 could inhibit the growth of MDR PA. Hence it can be concluded that silver nanoparticles synthesized in the presence of 5% gelatin can act as a bactericidal agent in the inactivation of MDR PA biofilms, thereby controlling the infections caused by these biofilms.

Strong immune responses and protection of PcrV and OprF-I mRNA vaccine candidates against Pseudomonas aeruginosa

Pseudomonas aeruginosa (PA) is a leading cause of hospital-acquired and ventilator-associated pneumonia. The multidrug-resistance (MDR) rate of PA is increasing making the management of PA a global challenge. Messenger RNA (mRNA) vaccines represent the most promising alternative to conventional vaccines and are widely studied for viral infection and cancer immunotherapy while rarely studied for bacterial infections. In this study, two mRNA vaccines encoding PcrV– the key component of the type III secretion system in Pseudomonas and the fusion protein OprF-I comprising outer membrane proteins OprF and OprI were constructed. The mice were immunized with either one of these mRNA vaccines or with the combination of both. Additionally, mice were vaccinated with PcrV, OprF, or the combination of these two proteins. Immunization with either mRNA-PcrV or mRNA-OprF-I elicited a Th1/Th2 mixed or slighted Th1-biased immune response, conferred broad protection, and reduced bacterial burden and inflammation in burn and systemic infection models. mRNA-PcrV induced significantly stronger antigen-specific humoral and cellular immune responses and higher survival rate compared with the OprF-I after challenging with all the PA strains tested. The combined mRNA vaccine demonstrated the best survival rate. Moreover, the mRNA vaccines showed the superiority over protein vaccines. These results suggest that mRNA-PcrV as well as the mixture of mRNA-PcrV and mRNA-OprF-I are promising vaccine candidates for the prevention of PA infection.

The impact of biosynthesized ZnO nanoparticles from Olea europaea (Common Olive) on Pseudomonas aeruginosa growth and biofilm formation

There is a limitation in the range of effectual antibiotics due to the Pseudomonas aeruginosa (PA) infection due to its innate antimicrobial resistance. Researchers have therefore been concentrating their efforts to discover advanced and cost effective antibacterial agents among the ever-increasing PA bacterial resistance strains. It has been discovered that various nanoparticles can be employed as antimicrobial agents. Here, we evaluated the antibacterial properties of the Zinc Oxide nanoparticles (ZnO NPs), which was biosynthesized, being examined on six hospital strains of PA alongside a reference strain (ATCC 27853). A chemical approach was applied to biosynthesize the ZnO NPs from Olea europaea was performed, and confirmed by using X-ray diffraction and Scanning Electron Microscopes. The nanoparticles then applied their antibacterial properties to examine them against six clinically isolated PA strains alongside the reference strain. This process tested for the results of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The Growth, biofilm formation and eradication were analyzed. The influence of the differentiating degrees ZnO NPs in regard to Quorom sensing gene expression were further examined. The ZnO NPs exhibited a crystalline size and diameter (Dc) of 40–60 nm and both the MIC and MBC tests revealed positive outcomes of concentrations of 3 and 6 mg/ml for each PA strain, respectively. At sub inhibitory concentration, The ZnO NPs were found to significantly inhibit the growth and biofilm formation of all PA strains and decreases in the biomass and metabolic behavior of PA established biofilms; these decreases varied depending on the dosage. At ZnO NPs concentrations of 900 µg/ml, the expression of majority of quorum sensing genes of all strains were significantly reduced, at ZnO NPs concentrations of 300 µg/ml, few genes were significantly impacted. In conclusion, the treatment of PA and could be other antibiotic resistant bacteria can therefore be approached by using ZnO NPs as it has been uncovered that they withhold advanced antibacterial properties.

Enhanced green luminescence properties of cu doped ZnO nano-flowers and their improved antibacterial activities

The copper doped ZnO (CuxZn1-xO, where x = 0, 0.01, 0.02, 0.04, 0.05) nanoflowers were produced by using co-precipitation process. Hexagonal wurtzite nanocrystalline phase of both pure and Cu-doped samples was confirmed by X-ray diffraction analysis. Field emission scanning electron microscopy results confirmed the emergence of fully dispersed nanoflower like morphologies in the Cu-doped ZnO and uniform dispersion of particles. Band gap energies of the prepared samples were calculated using the Tauc?s plot. All of the generated samples? room temperature photoluminescent spectra show two emission peaks, one excitonic peak in the UV region and another broad defect level peak in the visible area. The yellow and green luminescence characteristics of both pure and doped samples were analysed via the PL spectra in visible light region, making them appropriate for optoelectronic applications. The antibacterial potential of the produced nanostructures against the bacterial strains of Enterococcus Faecalis (gram positive) and Pseudomonas Aeruginosa (gram negative) were assessed. When compared to the pure ZnO nanostructures, the doped samples exhibit enhanced antibacterial activity. For the samples with 5 at.%Cu, the greatest zone of inhibition against Pseudomonas Aeruginosa was noted to be 25mm. For the samples doped with 5 at.% Cu, the largest zone of inhibition against Enterococcus Faecalis was 21mm. The findings indicate that the obtained samples have higher antibacterial potential against gram negative than gram positive bacteria. The prepared doped ZnO nanoflowers were therefore recommended as excellent green nanophosphors for the production of white LEDs and effective nanoantibiotics against gram-negative bacteria.

Iron Acquisition Proteins of Pseudomonas aeruginosa as Potential Vaccine Targets: In Silico Analysis and In Vivo Evaluation of Protective Efficacy of the Hemophore HasAp.

Pseudomonas aeruginosa (PA) is a Gram-negative pathogen responsible for fatal nosocomial infections worldwide. Iron is essential for Gram-negative bacteria to establish an infection. Therefore, iron acquisition proteins (IAPs) of bacteria are attractive vaccine targets. A "Reverse Vaccinology" approach was employed in the current study. Expression levels of 37 IAPs in various types of PA infections were analyzed in seven previously published studies. The IAP vaccine candidate was selected based on multiple criteria, including a high level of expression, high antigenicity, solubility, and conservation among PA strains, utilizing suitable bioinformatics analysis tools. The selected IAP candidate was recombinantly expressed in Escherichia coli and purified using metal affinity chromatography. It was further evaluated in vivo for protection efficacy. The novel immune adjuvant, naloxone (NAL), was used. HasAp antigen met all the in silico selection criteria, being highly antigenic, soluble, and conserved. In addition, it was the most highly expressed IAP in terms of average fold change compared to control. Although HasAp did excel in the in silico evaluation, subcutaneous immunization with recombinant HasAp alone or recombinant HasAp plus NAL (HasAP-NAL) did not provide the expected protection compared to controls. Immunized mice showed a low IgG2a/IgG1 ratio, indicating a T-helper type 2 (Th2)-oriented immune response that is suboptimal for protection against PA infections. Surprisingly, the bacterial count in livers of both NAL- and HasAp-NAL-immunized mice was significantly lower than the count in the HasAp and saline groups. The same trend was observed in kidneys and lungs obtained from these groups, although the difference was not significant. Such protection could be attributed to the enhancement of innate immunity by NAL. We provided a detailed in silico analysis of IAPs of PA followed by in vivo evaluation of the best IAP, HasAp. Despite the promising in silico results, HasAp did not provide the anticipated vaccine efficacy. HasAp should be further evaluated as a vaccine candidate through varying the immunization regimens, models of infection, and immunoadjuvants. Combination with other IAPs might also improve vaccination efficacy. We also shed light on several highly expressed promising IAPs whose efficacy as vaccine candidates is worthy of further investigation.

The Antibiotic Resistance Patterns of Pseudomonas Aeruginosa Strains Isolated from Microbiological Specimens Experiences of A Single Center on the Pseudomonas Aeruginosa Strains

Purpose To determine the antibiotic resistance rates of Pseudomonas aeruginosa strains isolated from samples collected by the microbiology laboratory of a single center for four years. Methods The samples of wound, blood, tracheal aspirate, abscess, vagina, cerebrospinal fluid, sputum, and urine culture isolated from 789 patients who were hospitalized in the service, intensive care and outpatient services between 2017-2021 were evaluated retrospectively for Pseudomonas species. Results Most of culture were urine (42.7%) and sputum cultures (20.4%). Most patients applied to the service were from department of chest diseases (38.6%) or from department of urology (14.3%) or palliative care unit (12.5%). The highest resistances were against cefuroxime, levofloxacin and netilmicin; lowest resistance was against amikacin. The aztreonam, cefepime and gentamicin resistances were significantly reduced by years (P=0.0321, 0.0038 and 0.0004, respectively) while colistin and levofloxacin resistance considerably increased (P&lt;0.0001 and P=0.0407, respectively). Significant decreases were observed in resistance against cefepime, ceftazidime and ciprofloxacin over years (P=0.0321, 0.0038 and 0.0004, respectively). A significant decrease in resistance of strains isolated from urine culture was only observed forcefepime over years (P=0.0003). The resistance of strains isolated from cultures of sputum, urine and respiratory secretions against levofloxacin significantly increased in 2019 while those of wound culture increased in 2020 (P=0.0145). Conclusion Alterations in the antibiotic resistance profile were detected in patients over years due to frequently varied use of antimicrobials. To select effective treatment protocols, it is necessary to regularly determine the microorganisms and their antibiotic resistance for each hospital.

Distantly Related Homologue of UhpT in Pseudomonas aeruginosa

Pseudomonas aeruginosa (PA) is an opportunistic Gram-negative bacteria that affects patients in intensive care units and chronic respiratory disease patients. Compared to other bacteria, it has a wide genome (around 6.3-Mb) that supports its metabolic versatility and antimicrobial resistance. Fosfomycin (FF) is primarily used as an oral treatment for urinary tract infections (UTIs). FF diffuses inside the cell via glycerol-3-phosphate transporter (GlpT) PA, as well as in other bacteria. In other bacteria, such as E. coli, glucose-6-phosphate transporter (UhpT) functions as FF transporter. Since mutant GlpT leads to FF resistant PA, it is assumed that GlpT is the only FF transporter. However, it is also assumed that PA uses glucose-6-phosphate and, thus, homologous proteins of UhpT may be present in its genome. Here, we present an attempt to find a distant related homologue of UhpT in PA. A Hidden Markov Model (HMM) was created to seek for Major facilitator family (MFS) domain in 21 PA genomes of 14 CF patients annotated with prokka and the statistical analysis was performed (MCC: 0.84, ACC: 0.99). Then, the HMM was applied to PA genomes. Besides the actual GlpT, annotated as glpt_1, one more GlpT protein was found in 21 out of 21 genomes, annotated as glpt_2. Since glpt_2 clusters closer to UhpT than GlpT, glpt_2 was selected to build a model. Computing a structural superimposition, the model and the template of UhpT have 0.6 Å of RMSD. The model of glpt_2 has some characteristics that are fundamental to UhpT functions. The binding site, consisting of 2 arginines (Arg46 and Arg275) and Lys45, is totally conserved, as well as the topology of the structure. Asp90 is also conserved in glpt_2 model. No studies aimed at searching for distant related homologous of UhpT. Since the high genetic exchange and high mutational rate in bacteria, it is likely that PA has a UhpT-like protein in the PA genome. The binding site is superimposable to UhpT protein as well as the overall topology. In fact, the 12 TMs are completely comparable, suggesting a well-defined folding of the protein across the bilayer lipid membrane. To enforce our hypothesis, in all 21 PA genomes, we also found a protein annotated as membrane sensor protein UhpC, important for expression and function of UhpT in E. coli. Since PA strains are wild-type, we can assume that most of the PA have proteins like this. The presence of a homologue of UhpT suggests that this protein is conserved in PA genome.

Synergistic Effect of Probiotic Bacteria and Antibiotics on Antibiotic-resistant Strains of Pseudomonas Aeruginosa Isolated from Patients with Burn Wounds

Synergistic Effect of Probiotic Bacteria and Antibiotics on Antibiotic-resistant Strains of Pseudomonas Aeruginosa Isolated from Patients with Burn Wounds