Microbiological and molecular studies on a multidrug-resistant Pseudomonas aeruginosa from a liver transplant patient with urinary tract infection in Egypt.

BACKGROUND: Pseudomonas aeruginosa is an opportunistic pathogen responsible for complicated UTIs and exhibits high antibiotic resistance, leading to increased mortality rates, especially in cases of multidrug-resistant strains. This study aimed to investigate the antibiotic susceptibility patterns and genomic characterization of XDR strains identified in end-stage liver disease patients who underwent liver transplants. METHODS: In this study, a number of 30 individuals who underwent liver transplants were registered. Ninety urine and 60 wound site swab samples were collected and processed for culturing, identification, and antimicrobial sensitivity. Extensively drug-resistant strain EMARA01 was confirmed through Sanger sequencing and was then processed for whole genome sequencing to characterize the genomic pattern. Sequencing data were processed for de novo assembly using various tools and databases, including genome annotation, serotype identification, virulence factor genes, and antimicrobial resistance gene. Pangenome analysis of randomly selected 147 reference strains and EMAR01 sequenced strain was performed using the Bacterial Pan Genome Analysis (BPGA) software. RESULTS: Of these total examined samples, nosocomial infection due to P. aeruginosa was detected in twelve patients' samples. AST analysis showed that P. aeruginosa strains exhibit resistance to tobramycin, erythromycin, and gentamicin, followed by piperacillin and ofloxacin, and no strains exhibit resistance to meropenem and imipenem. The CARD database identified 59 AMR genes similar to the EMAR01 strain genome and mostly belong to the family involved in the resistance-nodulation-cell division (RND) antibiotic efflux pump. Five genes; nalC, nalD, MexR, MexA, and MexB, exhibit resistance to 14 classes of antibiotics, while two AMR; CpxR, and OprM, exhibit resistance to 15 classes of drugs. Pangenome analysis revealed that the pan-genome remained open, suggesting the potential for acquiring accessory and unique genes. Notably, the genes predominantly involved in amino acid transport metabolism were identified using the KEGG database. CONCLUSIONS: This study provides valuable insights into the antimicrobial resistance profile, genetic features, and genomic evolution of P. aeruginosa strains causing UTIs in liver transplant patients. The findings emphasize the significance of comprehending AMR mechanisms and genetic diversity in P. aeruginosa for developing effective treatment strategies and infection control measures.

Antibacterial, antibiofilm, and anticancer activity of silver-nanoparticles synthesized from the cell-filtrate of Streptomyces enissocaesilis.

Silver nanoparticles (Ag-NPs) have a unique mode of action as antibacterial agents in addition to their anticancer and antioxidant properties. In this study, microbial nanotechnology is employed to synthesize Ag-NPs using the cell filtrate of Streptomyces enissocaesilis BS1. The synthesized Ag-NPs are confirmed by ultraviolet-visible (UV-Vis), Fourier transform infrared (FT-IR), X-ray diffraction (XRD), energy dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Also, the effects of different factors on Ag-NPs synthesis were evaluated to set the optimum synthesis conditions. Also, the antibacterial, antibiofilm, and anticancer activity of Ag-NPs was assessed. The X-ray diffraction (XRD) analysis confirmed the crystalline nature of the sample and validated that the crystal structure under consideration is a face-centered cubic (FCC) pattern. The TEM examination displayed the spherical particles of the Ag-NPs and their average size, which is 32.2 nm. Fourier transform infrared spectroscopy (FTIR) revealed significant changes in functionality after silver nanoparticle dispersion, which could be attributed to the potency of the cell filtrate of Streptomyces enissocaesilis BS1 to act as both a reducing agent and a capping agent. The bioactivity tests showed that our synthesized Ag-NPs exhibited remarkable antibacterial activity against different pathogenic strains. Also, when the preformed biofilms of Pseudomonas aeruginosa ATCC 9027, Salmonella typhi ATCC 12023, Escherichia coli ATCC 8739, and Staphylococcus aureus ATCC 6598 were exposed to Ag NPs 50 mg/ml for 24 hours, the biofilm biomass was reduced by 10.7, 34.6, 34.75, and 39.08%, respectively. Furthermore, the Ag-NPs showed in vitro cancer-specific sensitivity against human breast cancer MCF-7 cell lines and colon cancer cell line Caco-2, and the IC50 was 0.160 mg/mL and 0.156 mg/mL, respectively. The results of this study prove the ease and efficiency of the synthesis of Ag-NPs using actinomycetes and demonstrate the significant potential of these Ag-NPs as anticancer and antibacterial agents.

Antibacterial Potential of Bacterial Cellulose Impregnated with Green Synthesized Silver Nanoparticle Against S. aureus and P. aeruginosa.

In this study, bacterial cellulose (BC) impregnated with green synthesized silver nanoparticles (AgNPs) is evaluated as an antimicrobial membrane for wound-healing treatment. Green synthesized silver nanoparticles using Moringa oleifera leaf extract were characterized using UV‒visible spectroscopy, FTIR, X-ray diffraction, and transmission electron microscopy. The results confirmed that the resulted particles were Ag2O and metallic Ag in nanoscale with an average size ranged from 24 to 40 nm. The green synthesized nanoparticles incorporated within both bacterial cellulose and filter paper discs showed excellent antibacterial activities against Staphylococcus aureus ATCC 6538 and Pseudomonas aeruginosa ATCC 9027. There was no significant difference noticed between bacterial cellulose and filter paper holding capacity to nanoparticles and there was lack of interaction between bacterial cellulose and impregnated nanoparticles as elaborated by Fourier transform infrared spectral analyses. Scanning electron microscopy investigation showed major distortions effects of green synthesized silver nanoparticles on bacterial cell morphology.

Evaluation of a new antimicrobial agent production (RSMM C3) by using metagenomics approaches from Egyptian marine biota.

Diseases and epidemics in the current days need new types of antibiotics in order to be able to eliminate them. The goal of this research is to use metagenomics to identify isolated utilitarian gene (s) as antimicrobial specialists. Collection of diverse locations from sea sediment samples from Alexandria and extraction of total DNA, restriction enzyme fragmentation, cloning into pUC19 vector, and expression of the isolated gene(s) in E. coli DH5α were all part of the process. Characterization of Antimicrobial agent was done for the best clone for antimicrobial agent's production to detect efficiency, optimum pH, thermal stability, pH stability, effect of different compounds on antimicrobial activity, and residual activity of product after preservation in room temperature. Amino acid sequence of RSMM C3 gene (1250 bp) was 72% identity with Herbaspirillum sp. The ideal temperature level of the RSMM C3 antimicrobial agent production was 36 °C. The antimicrobial agent RSMM C3's stability was stable at -20 °Celsius for up to two months without thawing. The antibacterial agent RSMM C3 was stable at 4 °C for 14 days without loss in activity. The ideal pH level of the RSMM C3 antimicrobial agent was 6. Remain activity was gradually decreased at pH 5, 6, 6.5 and 7 (86.1, 96.9, 97.2 and 94.9%, respectively). On the other hand, residual activity was (92 and 84%) at (pH 7.5 and 8) for 8 days. The tested antimicrobial RSMM C3 was stable against 1 mM of different compounds (DMSO, Glycerol, NaCl, CaCl2, MgCl2, ZnCl2, FeSO4, MnSO4 and CuSO4). The research provides for the Metagenomics technique that has the ability for the production of novel antimicrobial agents produced by clone RSMM C3 which has a wide spectrum activity towards different microorganisms comparing to other antibiotics as Ampicillin and Tetracycline.

Antibacterial activities of hexadecanoic acid methyl ester and green-synthesized silver nanoparticles against multidrug-resistant bacteria.

Antibacterial drug resistance is considered one of the biggest threats to human health worldwide, and the overuse of antibiotics accelerates this problem. Multidrug-resistant (MDR) bacteria are becoming harder to treat as the antibiotics used to treat them become less effective. Therefore, it is necessary to evaluate novel methods to control MDR bacteria. In this study, 40 bacterial isolates were collected from diabetic patients. The sensitivity of 40 bacterial isolates to seven antibiotics was evaluated. Four bacterial isolates were resistant to all antibiotic groups. The MDR pathogenic bacteria were selected and identified morphologically and biochemically and confirmed by VITEK® 2 system as follows: Staphylococcus aureus W35, Pseudomonas aeruginosa D31, Klebsiella pneumoniae DF30, and K. pneumoniae B40. Identification of the most resistant P. aeruginosa D31 was confirmed by the sequencing of a 16S ribosomal RNA gene with an accession number (MW241596). The inhibitory activity of eight types of native grown plant extracts against MDR bacteria was studied. Clove alcoholic extract (CAE) showed the highest inhibitory activity against MDR bacteria. Gas chromatography-mass spectrometry analysis of partially purified CAE at 0.9 Rf detected by thin-layer chromatography showed an active compound named hexadecenoic acid methyl ester with the highest antimicrobial effect against clinical pathogenic bacteria. The formation of silver nanoparticles (AgNPs) by CAE was studied. Evaluation of AgNPs was investigated by X-ray diffraction, UV-Vis, and transmission electron microscopy. The antibacterial effect of AgNPs after 2, 4, and 6 days in light and dark conditions was evaluated. Finally, the AgNPs synthesized using CAE possess good inhibition activity against the tested pathogenic bacteria. As a result, the bactericidal components listed above were promising in reducing MDR bacteria and can be used for treatments of bacterial infection and in the development of safe products with a natural base.

Bifidobacterium longum Suppresses Murine Colorectal Cancer through the Modulation of oncomiRs and Tumor Suppressor miRNAs.

The modulatory role of the Bifidobacterium longum (BL), isolated from women breast milk, on some oncogenic and tumor suppressor miRNAs as well as IL-1ß and IL6 targeted-miRNAs was investigated using murine colorectal cancer (CRC) induced on the top of inflammatory ulcerative colitis model. The investigation of the oncomiRs miR-21a and miR-155, which regulate IL-6 and IL-1ß expression, indicated that both was depressed by BL-administration in healthy and in CRC-mice. BL-administration induced the tumor suppressor miRNAs (miR-145 and miR-15a) expression in both of the healthy and in CRC-mice. The miR-146a expression, which regulates both of IL-1ß and IL-6 expression, was decreased after the BL-administration in both of the healthy and in CRC-mice. In CRC-mice, NF-Kb concentration was elevated, however this NF-Kb induction was diminished after the treatment with BL. BL highly enhanced the IL-1ß and IL-6 mRNA and protein concentrations in healthy mice. The administration of BL to CRC-mice resulted in a dramatic increase in IL-1ß mRNA and IL-1ß concentration, which in contrast was accompanied with a decrease in the IL-6 mRNA and IL-6 concentration. BL-administration resulted in a drop in the aberrant crypt foci number in CRC-mice and increased necrosis and fibrosis of the colon cells. The modulatory influence of B. longum on microRNAs may provide an important therapeutic impact in CRC through inhibition of the proliferation, invasion, apoptosis, and cell cycle of tumor cells.

Macroalgal activity against multiple drug resistant Aeromonas hydrophila: A novel treatment study towards enhancement of fish growth performance.

OBJECTIVE: The aim of this study was to evaluate the efficiency of macroalgal extracts as antibacterial agent against multidrug-resistant (MDR) bacteria isolated from Nile tilapia (Oreochromis niloticus) as well as to enhance the fish growth performance by macroalgae diet application. METHODS: A total of 50 swabs were collected from the diseased organs of tilapia fish including gills, skin, spleen, intestine, liver, kidney and muscle. The isolated bacteria were identified and then confirmed by using VITEK 2. Eight macroalgal species were collected from Abu-Qir, Alexandria coast, Egypt. After determination of their biomass, three solvents were used to prepare algal extracts. The antibacterial activities of different macroalgal extracts were measured against MDR Aeromonas hydrophila 6 (MDRAH6) using well-diffusion method. The mechanism by which macroalgal extract affects MDR bacteria was conducted by using transmission electron microscope (TEM). To evaluate the safety of the promising algal extract, GC-MS was performed to detect the composition of S. vulgare extract. In addition, growth performance was measured as an application of algal extracts into fish feed. RESULTS: Between eight collected macroalgal species, Sargassum vulgare showed the highest biomass production (53.4 g m-2). In addition, its ethanolic extract showed the highest significant antibacterial activity with MIC value of 250 µg ml-1. TEM examination showed distinctive changes in the treated MDRAH6 cells including rupture of the cell wall, leakage of cytoplasmic contents, alterations in the cytoplasm density in addition to totally cell deformation. In addition, GC-MS analysis revealed eleven identified components in S. vulgare ethanolic extract, in which 9,12-octadecadienoyl chloride and hexadecanoic acid methyl ester were dominant (46.6 and 19.7 %, respectively). Furthermore, dietary replacement of fish meal with S. vulgare ethanolic extract significantly enhanced the growth performance and survival of Nile tilapia with a significant reduction in the total bacterial count. CONCLUSION: Ethanol extract of the brown macroalga S. vulgare could be a promising antibacterial and a new active agent against MDR A. hydrophila, which could be a major causative agent of Nile tilapia fish diseases. In addition, this study recommended S. vulgare as a natural and effective source to enhance the growth performance of Nile tilapia. In fact, isolation and examination of the individual antibacterial active compounds of the S. vulgar ethanolic extract are under investigation.

Antimicrobial activity of new 2,4-disubstituted thiazolidinone derivatives.

A number of new disubstituted 2,5-thiazolidinone derivatives were synthesized and tested for their antimicrobial activity against Bacillus subtilis (Gram-positive), Pseudomonas aeruginosa (Gram-negative), and Streptomyces species (Actinomycetes). They displayed different degrees of antimicrobial activities or inhibitory actions.