Globally, there is a growing concern about Pseudomonas aeruginosa resistance to aminoglycosides. Enzymatic modification of these drugs is the predominant resistance mechanism. Additionally, P. aeruginosa possesses many efflux systems that enable it to resist a variety of antimicrobial agents. This study aimed to determine the resistance patterns to several antibiotics as well as the possible mechanisms of aminoglycosides resistance, including aminoglycosides modifying enzymes (AMEs), genes, and active efflux system, observed in clinical P. aeruginosa isolates recovered from patients admitted to Minia University Hospitals, Minia, Egypt. Isolates of P. aeruginosa were identified by traditional phenotypic tests and assessed for their in vitro susceptibility to various antibiotics. The minimum inhibitory concentrations (MICs) of some aminoglycosides were determined without and after the addition of carbonyl cyanide m-chlorophenylhydrazone (CCCP). Aminoglycoside resistance amplified gene sequences were detected using polymerase chain reaction (PCR). Antibiotic sensitivity testing was applied on 93 clinical isolates of P. aeruginosa. The highest rate of resistance was recorded against cefepime and ceftazidime (94.6 % each), while 35.5 % of the examined strains exhibited resistance to minimally one of the evaluated aminoglycoside antibiotics. Furthermore, 49.5 % of isolates were multidrugresistant (MDR). After CCCP addition, 24.2 % of the resistant isolates restored their sensitivity to gentamicin. According to PCR analysis, aac(3)-II was the most frequently detected gene (21.2 %) followed by aph(3′)-VI (15.2 %), and aac(6’)-IIa (3 %). Multiple drug resistance was observed among P. aeruginosa strains included in this study. Resistance of P. aeruginosa to aminoglycosides is greatly influenced by efflux pumps. Coordinated measures and further investigations are urgently needed to manage aminoglycosides resistance.

Resistance to antimicrobial agents is a developing issue that requires calls for an integrated strategy from all nations. A significant contributor to antimicrobial resistance (AMR) is wastewater that is a vital resource for bacteria which provides a medium for gene transfer. Stressors like high concentration of hydrogen ions, antibiotics, and minerals can start and spread AMR in wastewater. Antimicrobial agents are present in the environment in varying amounts depending on the antimicrobial class and their frequent use. Evolution of antibioticresistant bacteria (ARB) poses a significant public health risk; especially in healthcare facilities and hospital's wastewater. Advanced wastewater treatment technologies should be implemented for effective treatment of hospital wastewater (HWW). Standardized phenotypic approaches are used to detect AMR in bacteria. Molecular approaches are now preferred to be used in laboratories instead of phenotypic approaches as they are faster and more accurate for detection of AMR's underlying genetic mechanisms. The most common molecular approaches for assessing AMR in water include quantitative polymerase chain reaction (PCR) and whole genome sequencing (WGS). Metagenomic WGS sequencing provides more extensive genomic data and taxonomic classification than quantitative PCR, as it makes sequencing for the entire genome of microorganisms. The objectives of this review were to demonstrate the primary mechanisms through which bacteria develop resistance to antimicrobials in water, and investigate the impact of hospital effluent water on the spread AMR and ARB. Additionally, various approaches for assessing AMR levels and guidelines for preventing and managing the transmission of AMR to the environment were discussed.

Escherichia coli and Klebsiella pneumonia; along with other Gram-negative bacteria (GNB), are known for their resilience to a broad spectrum of antibiotics, including last-resort options such as colistin, which is regarded as the last line of defense in treating multidrugresistant (MDR) infections. This elevated level of resistance presents a substantial threat to the healthcare system, paving the way for a post-antibiotic era. In the present study, a checkerboard assay was employed to pinpoint the optimal combinations of Origanum majorana (OM) essential oil and colistin (CS) for the purpose of enhancing colistin's antibacterial and antibiofilm activities (FICI ≤ 0.5). The investigation focused on eight colistin-resistant (CS_R) bacterial isolates, with a particular focus on two strains carrying the mobile colistin resistance mcr-1 gene. The CS@OM combination resulted in a 2-to 16-fold improvement of colistin efficacy, resulting in a biofilm eradication rate ranging from ∼ 52 % to 82 %. Furthermore, it evinced a complete eradication of the free-floating cells of the isolates, inclusive of those harboring the mcr-1 gene. The bactericidal activity of the CS@OM combination was achieved through various mechanisms, including bacterial membrane disruption and leakage of internal bacterial constituents, leading to bacterial death. Consequently, the CS@OM combination evinced a high capacity to prevent the development of drug resistance in the examined isolates. To the best of our knowledge, this is the first study to appraise the combination of colistin and Origanum majorana essential oil against colistinresistant isolates, including those carrying mcr-1 gene with the aim of regaining colistin susceptibility.

Plant growth promoting rhizobacteria (PGPR) have evolved to thrive in challenging environments and provide the plants with defense against the harmful impacts of the environmental stressors. The present study aimed to isolate, screen, and identify PGPB obtained from the rhizosphere soil of some field crops at different sites in Ismailia, Egypt. Isolation process was carried out from drought-suffered locations to obtain efficient promising strains adapted to carry out vital processes under an irrigation water shortage. Eight bacterial isolates were selected and identified by phenotypic properties, and were subjected to screening procedures to assess their growth capabilities and evaluate their potential as PGPB. The screening process involved investigating various PGPB features, such as phosphate solubilization, formation of indole-3-acetic acid (IAA), ammonia, and hydrogen cyanide (HCN) production. Among the eight isolates, two isolates only (MW3 and AB3) gave positive results for all the tested plant growth promoting traits. The promising isolates were identified by sequencing of their16S rRNA as Arthrobacter globiformis (MW3) and Micrococcus luteus (AB3). A field trial was conducted to evaluate the activity of the two PGPB and their mixture to act as biofertilizers for maize under deficit irrigation 0.75 from crop evapotranspiration (ETc). All tested inoculants significantly increased yield components of maize, NPK uptake by plants, availability of N and P in soil, activity of some soil enzymes, and total bacterial counts compared to the un-inoculated control. Utilization of stress adapting PGPR showed great potential in overcoming the challenges of sustainable agriculture under environmental stress conditions.

This research aimed to assess the metabolic activities of Azotobacter vinelandii DT1 strain by assaying its alginate synthesizing enzymes and modifying genes, synthesize alginate from the agricultural residues, and eradicate the pathogenicity and risk associated with Pseudomonas aeruginosa alginate. Preliminary screening for alginate production revealed the presence of different alginate products. High-performance liquid chromatography (HPLC) confirmed the existence of varying alginate concentrations, such as sodium alginate (2.42754×10-1 g/ 100 ml), calcium alginate (1.09597×10-1 g/ 100 ml), acid alginate (1.39420×10-2 g/ 100 ml), alginate oligosaccharide (8.20576×10-2 g/100 ml), and potassium alginate (9.78836×10-2 g/ 100 ml). These were accompanied with the corresponding alginate synthesizing enzymes; mainly GDP-Mannose dehydrogenase (23.77± 0.13 U/ ml); glycosyltransferase (9.68± 0.53 U/ ml), phosphomannomutase (266.09± 0.16 U/ ml), mannose phosphate isomerase (95.87± 0.51 U/ ml), alginate lyase (24.50± 0.95 U/ ml), and mannuronan epimerase (49.93± 0.82 U/ ml). In this study, the expression of alginate-modifying genes such as alginate lyase and GDP-Mannose dehydrogenase amplicons of Azotobacter vinelandii DT1 strain corresponding to 766 bp, 43 ng and 600 bp, 33 ng molecular weights of the fast DNA marker; justified the synthesis of different alginate products. Azotobacter alginate synthesis using a low-cost substrate (i.e., corn cobs) and completely non-pathogenic bacteria (Azotobacter vinelandii DT1) may be desirable compared to the pathogenicity risks and poor jellying qualities linked to Pseudomonas alginate biosynthesis, its expensive production costs, and the adverse environmental effects of seaweed harvesting and processing.

The escalating incidence of nosocomial infections stemming from ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) bacterial pathogens presents a formidable clinical challenge globally, affecting both the developed and developing nations. These pathogens, which are distinguished by their robust antibiotic resistance mechanisms, pose a significant threat to the public health. Their ability to evade traditional antimicrobial treatments underscores the urgent need for novel therapeutic stratifies or alternative approaches to mitigate their negative impact. Understanding the intricate mechanisms underpinning antibacterial resistance in ESKAPE bacteria is paramount for developing effective interventions. Enhanced insights into these mechanisms will facilitate the prediction of resistance patterns among the multidrug-resistant pathogens, thereby guiding the development of targeted therapies and preventive measures. Consequently, comprehensive efforts are needed aiming at unraveling the intricacies of antibacterial resistance in ESKAPE pathogens that are imperative to safeguarding the public health. The aim of the present review was to highlight the patho-mechanisms of ESKAPE bacteria towards the different antibiotics and genes involved in multi-drug resistance.

Bacillus paralicheniformis is a new identified species, which was distinguished from Bacillus licheniformis in 2015 through extensive phylogenomic and phylogenetic analyses. In this context, this study aimed to achieve a clear identification of the active plant-growth promoting rhizobacteria (PGPR) B. paralicheniformis isolates among the closely related B. licheniformis through molecular typing, helping for the development of clearly-identified PGPR isolates to be used as biofertilizers. A total of 15 rhizobacteria were isolated from the olive rhizosphere soils. These bacterial isolates exhibited various proprieties in terms of abiotic stress tolerance, biofilm formation under stress conditions, and enzyme activities (i.e., lipases, cellulases, and proteases). In addition, several PGP traits such as phosphate solubilization and the production of siderophores and indol-3 acetic acid were also observed. Molecular identification through 16S rRNA sequencing initially identified all the isolates as Bacillus spp. The multi-locus sequence typing (MLST) scheme, using six housekeeping genes (adk, ccpA, recF, rpoB, sucC, and spo0A) had unveiled distinct allelic profiles resulting in 13 unique sequence types (ST). Notably, a comprehensive analysis indicated no exact allele matches existed between the examined isolates and those documented in the PubMLST database. Among the six housekeeping genes, we noticed that rpoB gene (RNA polymerase, subunit beta) had multiple polymorphic sites that were bordered by conserved sequences.

Drechslera graminea is the causal agent of leaf-stripe disease in barley leading to substantial crop losses. The current work aimed to investigate the antagonistic activity of Trichoderma harzianum against D. graminea. Pathogen was isolated from the naturally infected barely leaves and molecularly identified. The identified isolate was deposited in GenBank with an accession number of OR827023. In vitro dual culture assay showed a decrease in D. graminea radial growth with a growth inhibition percentage of 77.8 %. T. harzianum culture filtrate (TCF) (30×10 6 spore/ ml) was efficiently applied in the greenhouse causing a significant reduction in final disease severity (FDS %) by 77 %, compared to the positive control. Moreover, application of TCF against the natural infection in the open field reduced FDS, the area under the disease progress curve (AUDPC), and the average coefficient of infection (ACI). Furthermore, TCF significantly improved yield components compared to the untreated control. To assess the plant response on a molecular basis, the protein profile was analyzed showing a wide range of variability. Five polymorphic bands were recognized; mainly 66, 48, 35, 29, and 16 kDa. The band with a molecular weight of 66 kDa had completely disappeared in the negative control, but was observed in the positive control and upon application of TCF. Meanwhile, the bands with molecular weights of 35, 29, and 16 kDa were recorded only in the positive control but were completely vanished in the negative control and upon application of TCF. Additionally, upon application of TCF under the infection stress, phenylalanine ammonia lyase (PAL) mRNA transcript accumulation had increased significantly in a way that exceeded two-fold of the non-infected control. Finally, it was concluded that T. harzianum was able to reduce leaf-stripe infection damage and promoted the barley-induced resistance in the susceptible cultivars; with the improvement of vegetative growth parameters, which may be a real prospect for leaf-stripe disease management in the future.

The aim of this study was to evaluate the genetic diversity of some Zucchini yellow mosaic virus (ZYMV)-infected cucurbits compared to the healthy plants based on three molecular tools. When sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis was used to determine the genetic variability of four cucurbits infected with ZYMV compared to the healthy ones, a total of 15 storable protein bands were recorded. The experimental results showed that DNA polymorphisms included 38 polymorphic DNA fragments that were generated using the five inter simple sequence repeat (ISSR-PCR) used primers. The maximum identity (%) among the ZYMV-infected and healthy samples was 59, 56, 45, 40, 30, and 29 % for the plant samples of watermelon, squash, cantaloupe, qethaa, cucumber, and luffa samples, respectively. Phylogenetic tree of DNA polymorphisms confirmed the genetic relationship between each of the healthy samples of watermelon and cucumber, healthy samples of qethaa and cucumber, ZYMV-infected samples of watermelon and squash, and ZYMV-infected samples of kantaloupe and qethaa. Numbers of nine start codon targeted (SCoT) primers were used to determine the DNA fingerprinting of six ZYMinfected cucurbit species compared to the healthy plants of the same species. A total of 88 polymorphic DNA fragments were distributed as follows: 13,

The present study aimed to evaluate the concepts of epidemiology, uropathogenic, diagnosis, prevention, and treatment of urinary tract infections (UTI) in humans. One of the most frequent infections that affect people is UTIs. During childhood they are equally common in boys and girls, and after that, they are more common in girls. In both of the general population and hospital environment, women frequently experienced at least one UTI in their lifetime. The existence of bacteriuria and pyuria are the 2 most significant signs of UTIs. Frequent urination, pain during urination, and soreness in the side or lower back are among the main symptoms of UTI. These infections are classified into 3 primary types: asymptomatic bacteriuria, lower UTI (cystitis), and febrile upper UTI (acute pyelonephritis), because such classification supports understanding of the infection"s etiology. UTI can be diagnosed through a combination of positive urine tests and/or culture and symptoms. Dipstick urinalysis is widely used due to its ease of availability and utility; however, the outcomes must be evaluated by considering the patient"s pretest probability depending on the characteristics and symptoms. Most UTIs can be treated with antibiotics such as Amoxicillin, cephalexin, and doxycycline. But the Gram-positive bacterium (Enterobacter faecalis) exhibited great resistance to erythromycin, while the Gram-negative bacterium (Escherichia coli) displayed great resistance to ampicillin. So, there is an urgent need for a combination between organic treatments and antibiotics to treat the complicated UTI. For example, the plant-based treatments, such as cranberry juice, are efficient in treating the UTI and can be used as an alternative to combating the bacteria that cause UTI.