Surface water contamination remains a major worldwide public health concern and may contribute to the dissemination of antibiotic-resistant bacteria. The Al-Hillah River in the city of Babylon Province, Iraq, diverts flows from the Euphrates River. Because of its importance in irrigation and population density, it faces several forced and unforced changes due to anthropogenic activities. To evaluate water quality, water samples were collected from three sites with different anthropogenic pressures along the Al-Hillah River. These samples were subjected to bacteriological analyses, i.e., total coliforms, Escherichia coli, and faecal enterococci. The phylogenetic groups of the E. coli isolates (n = 61) were typed by rapid PCR-based analyses. Representatives of each isolate were tested phenotypically for resistance to six classes of antibiotics and characterized according to their phylogenetic groups. The results demonstrated the highest resistance levels were to β-lactam antibiotics, followed by fosfomycin and aminoglycosides. Escherichia coli isolates belonging to phylogenetic groups A and B2 were the most common and were characterized by a higher prevalence of antibiotic resistance. This study is important for understanding the current conditions of the Al-Hillah River, as the data reveal a high prevalence of multiresistance among E. coli isolates circulating at the three sampling sites.
The present study aimed to isolate and identify constituents with antibacterial activity from the Methanol extract of Melastoma malabathricum leaves (MMML) through bioassay guided fractionation. Furthermore, the study scoped to evaluate the cytotoxic effects of M. malabathricum leaves fractions for possible protective effect of antoxidative constituents present in the fractions. The MMML extract was fractionated by Vacuum Liquid Chromatography (VLC) to afford M. malabathricum leaves fractions (ML1-ML6) of increasing polarities. Antibacterial activity of plant extract, six fractions and two bioactive constituents against Staphylococcus aureus reference strain, methicillin-resistant Staphylococcus aureus reference strain (MRSA), eleven clinical MRSA isolates, three clinical Pseudomonas aeruginosa isolates and P. aeruginosa reference strain was evaluated using Minimum Inhibitory Concentration (MIC). The bioactive constituents responsible for antibacterial activity of MMML, fractions were detected by direct TLC-bioautography. The data obtained from MIC assay showed that ML5 effectively inhibited growth of all test bacterial pathogens. Direct TLC-bioautography revealed that ML5 had the highest number of antibacterial compounds. Following bioassay-guided fractionation, Kaempferol-3-O-(2â, 6â-di-O-p-trans-coumaroyl)-β-glucopyranoside and Kaempferol were isolated from ML5. The structures of bioactive compounds were elucidated using Nuclear Magnetic Resonance (NMR) spectroscopy from 1HNMR and 13CNMR. In addition, the cytotoxic effect of M. malabathricum leaves fractions (ML1-ML6) revealed that ML5 had the highest Cytotoxicity Concentration (CC50) at 0.75 mg mL-1, an observation traceable to the presence of flavonoids constituents with antioxidant properties. Therefore, the results clearly indicate that MMML, fractions and bioactive constituents have profound antibacterial activity. Furthermore, antioxidative compounds from ML5 fraction exert their effects by enhancing the level of CC50
Pseudomonas aeruginosa is one of the main causes of nosocomial infections and is frequently associated with opportunistic infections among hospitalized patients. Kaempferol-3-O-(2′,6′-di-O-trans-p-coumaroyl)-β-D glucopyranoside (KF) is an antipseudomonal compound isolated from the leaves of the native medicinal plant Melastoma malabathricum. Herein, an RNA-seq transcriptomic approach was employed to study the effect of KF treatment on P. aeruginosa and to elucidate the molecular mechanisms underlying the response to KF at two time points (6 h and 24 h incubation). Quantitative real-time PCR (qRT-PCR) was performed for four genes (uvrD, sodM, fumC1, and rpsL) to assess the reliability of the RNA-seq results. The RNA-seq transcriptomic analysis revealed that KF increases the expression of genes involved in the electron transport chain (NADH-I), resulting in the induction of ATP synthesis. Furthermore, KF also increased the expression of genes associated with ATP-binding cassette transporters, flagella, type III secretion system proteins, and DNA replication and repair, which may further influence nutrient uptake, motility, and growth. The results also revealed that KF decreased the expression of a broad range of virulence factors associated with LPS biosynthesis, iron homeostasis, cytotoxic pigment pyocyanin production, and motility and adhesion that are representative of an acute P. aeruginosa infection profile. In addition, P. aeruginosa pathways for amino acid synthesis and membrane lipid composition were modified to adapt to KF treatment. Overall, the present research provides a detailed view of P. aeruginosa adaptation and behaviour in response to KF and highlights the possible therapeutic approach of using plants to combat P. aeruginosa infections.
Aquatic environment contamination remains a foremost global public health hazards, and symbolizes a significant reservoir of releasing antibiotic resistant bacteria. The survival of Escherichia coli in aquatic environments serves as a potential reservoir of antibiotic resistance, encompassing but not restricted to a plasmid-mediated quinolone resistance (PMQR) mechanism. The current study aimed to detect the presence of the PMQR-qnrA gene in quinolone-resistant E. coli isolates. Sixty-one waterborne E. coli with known phylogroups/subgroups isolated from the Al-Hillah River in Babylon Province, Iraq, were screened for the phenotypic resistance to third-generation quinolones (levofloxacin and ofloxacin) and were further analysed for the presence of the qnrA gene using polymerase chain reaction (PCR). Fifty-seven (93.4%) of 61 E. coli isolates were levofloxacin-resistant, and 55 (90.2%) were ofloxacin-resistant. Among the 57 quinolone-resistant E. coli, 40 (65.57%) isolates were found to carry the PMQR-qnrA gene. Among the 40 qnrA-positive E. coli, 22 (36.1%) isolates were in phylogroup B2, followed by 8 (13.1%) isolates in phylogroup D, 6 (9.8%) isolates in phylogroup B1, and 4 (6.6%) isolates in phylogroup A. The presence of the PMQR-qnrA gene in E. coli belonging to phylogroup B2 and D reflects the need for routine monitoring of antibiotic resistance genes (ARGs) in the Al-Hillah River.
Extended-spectrum ß-lactamases (ESBL) are a major source of concern. ESBL have been recorded around the world. Globally, the number of people infected with Enterobacteriaceae that produce extended-spectrum beta-lactamase (ESBL) is on the rise. It has been a rise in resistance to ß-lactam antibiotics among them. In this study, the objective was to collect Escherichia coli isolates from Urinary tract infection patients using selective medium, determine the prevalence of ESBL-producing E. coli, phylogenetic groupings of isolates, ESBL production, and biofilm formation among the isolates of E. coli isolates. The study included 250 E. coli samples from male and female subjects and grown on a selective medium. The isolated bacteria were submitted to different tests, including the detection of biofilm development and testing of the phylogenetic grouping of the E. coli isolate using triplex-PCR analysis. Representatives of each isolate were phenotypically evaluated for antibiotic resistance and classified into phylogenetic groupings. The results of extended-spectrum ß -lactams antibiotics showed the greatest resistance levels. There were 100% resistance rates for Ceftazidime-Clavulantae (CZC) and Cefotaxime-Clavulantae (CTC), 78.7% for Ceftazidime (CAZ), 86.7% for Cefotaxime CTX, 84% for Aztreonam (ATM), 87.3% for Ceftriaxone (CRO) and 83.3% for Cefpodoxime (CPD). E. coli isolates belonging to phylogroup B2 (91, 91%), and subtyping B23 (75, 75%) were the most common among UTI patients. ESBL-producing E. coli isolates were prevalent in individuals with UTIs. Most E. coli isolates from UTI patients at Al-Hillah hospitals belonged to phylogroup B2, followed by D, B1, and A. B2 was the most prevalent group in the study. This study examined the dissemination of ESBL genes in phylogenetic groups of the E. coli isolates from UTIs patients in the Al-Hillah, Iraq.
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