Broad-Spectrum Antimicrobial, Antioxidant, and Anticancer Studies of Leaf Extract of Simarouba glauca DC In Vitro.

The current study aimed to screen the preliminary phytochemicals in the leaf extract of the medicinal plant Simarouba glauca and to analyze its potential antimicrobial, antioxidant and anticancer properties. The phytochemical profile of the methanol extract was analyzed, and bioactive compounds were identified using chromatography, FTIR and GCMS. Antimicrobial activity and Minimum Inhibitory Concentration (MIC) were determined against 14 bacterial and 6 fungal strains. Moreover, the synergistic effect of a plant extract with commercially available antibiotics was also evaluated using the checkerboard method. The ethanolic and methanolic extracts showed exclusive activity against S. aureus and profound activity against E. coli and S. marcescens. Upon comparing breakpoints, methanolic extract demonstrated higher antimicrobial activity with a MIC value of 3.2 mg/mL against the test pathogens. Furthermore, the extracts demonstrated potential antioxidant activity; methanol extract had higher antioxidant potential compared to the ethanol extract. The major proactive bioactive compound with maximum antioxidant capacity was observed to be terpenoids. The methanol extract of S. glauca showed significant cytotoxicity against the MCF-7 breast cancer cell line with an IC50 value of 16.12 µg/mL. The overall results of our work provide significant evidence for the usage of methanolic extract of S. glauca as an efficient ethnomedicinal agent and a potential candidate for relieving many human ailments.

Comparative phenotypic characterization identifies few differences in the metabolic capacity between Escherichia coli ST131 subclones.

BACKGROUND: Extraintestinal pathogenic Escherichia coli (ExPEC) is responsible for causing many infections such as urinary tract infections (UTIs). The current dissemination of the multidrug resistant (MDR) ExPEC clone, Escherichia coli sequence type 131 (E. coli ST131), poses a real threat to public health worldwide. This study aimed to determine and compare the metabolic capacity of a collection of ExPEC isolates including ST131, non-ST131 and various ST131 subclones, and sought to assess the association between antimicrobial resistance and metabolic capacity of ST131 isolates. METHODS: The metabolic activity of forty urine E. coli isolates, collected from in-patients hospitalized at tertiary hospital in Riyadh, was tested using KB009 Hi carbohydrate kit, and then statistically analysed to assess the difference in the metabolic profiles between ST131 and non-ST131 isolates, and between ST131 subclones. RESULTS: The data of this study found almost similar metabolic profiles between ST131 and non-ST131, suggesting that ST131 is not a metabolically unique clone of ExPEC. There was also no link between antimicrobial susceptibility profiles and high metabolic capacity of ST131 isolates. Testing the biochemical activity of isolates belonging to ST131 subclones found higher activity of H30 subclone than non-H30 isolates, however it revealed few significant differences between these subclones. CONCLUSION: This study demonstrated no difference in the metabolism of ST131 and non-ST131, although it uncovered the presence of few significant differences in the metabolic capacity between ST131 subclones. Carrying out whole-genome based studies on ST131 and its main subclones is essential to elucidate the genetic factors responsible for the success of particular ST131 subclones.

Sericin-functionalized GNPs potentiate the synergistic effect of levofloxacin and balofloxacin against MDR bacteria.

A gradual expansion in resistant bacterial strains against commercially available antibacterial agents is the serious concern of the given research. It poses critical problem for public health. Thus, the demand for new antimicrobial agents has increased the interest in newer technologies and innovative approaches are required to advance the diagnosis and elimination of causative organisms. In this study, the potential role of technologies based on gold nanoparticles (GNPs) has been evaluated. GNPs were synthesized by using a cysteine protease, sericin whose reducing properties were exploited to bioengineer NPs (SrGNPs) where sericin with the help of thiol groups encapsulated over the surface of GNPs. Further, SrGNPs were bioconjugated with levofloxacin (Levo) and balofloxacin (Balo) to increase the efficacy of these drugs. Here, the antibacterial action of SrGNPs and their bioconjugated counterparts comprising Levo (Levo-SrGNPs), Balo (Balo-SrGNPs), and Levo/Balo (Levo-Balo-SrGNPs) were examined against normal and multi-drug resistant (MDR) strains of E. coli and S. aureus. The minimum inhibitory concentration (MIC) of these bioconjugates against said bacteria were found less than their pure counterparts. Further, the synergistic role of SrGNPs in combination with Levo and Balo was also explained using Chou-Talalay (C-T) method. The synthesis and bioconjugation of SrGNPs were confirmed by UV-visible spectroscopy, dynamic light scattering (DLS), transmission electron microscopy (TEM), and zeta-potential.

Incidence and antimicrobial profile of extended-spectrum ß-lactamase producing gram-negative bacterial isolates: An in-vitro and statistical analysis.

OBJECTIVES: There is an extensive incidence of extended-spectrum beta-lactamases (ESBLs), principally in the hospital environment across the world. The present study was designed to discover the frequency of ESBL-production among the clinical isolates of Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. The study also focused on determining their liability to the selected antimicrobials. METHODS: Two hundred ten (210) clinical specimens were tested for the occurrence of ESBL using the double-disc synergy test. The molecular, physicochemical, absorption, distribution, metabolism, excretion, and toxicity were checked through an online server. RESULTS: Among the screened clinical isolates, E. coli (n=44), K. pneumonia (n=34) and P. aeruginosa (n=14) were ESBL markers. The ESBL producing isolates exhibited co-resistance to diverse categories of antibiotics. It was observed that all the ESBL-producing isolates were sensitive towards imipenem and faropenem with minimal proportion of resistance. CONCLUSION: The imipenem and faropenem can be recommended as the drugs of selection due to a lesser amount of resistance as compared to other antibiotics in this study.

In-vitro antibacterial, antioxidant potentials and cytotoxic activity of the leaves of Tridax procumbens.

The present study explored the phytochemicals, antibacterial, antioxidant and cytotoxic effect of Tridax procumbens leaves. The leaves were dried and extracted with various organic solvents. The leaves contained the phytochemicals such as alkaloids, carbohydrates, polyphenols and tannins respectively. Antimicrobial potentials of the extracts were determined by performing the disc diffusion techniques. Results revealed that different organic solvents extracts namely methanol, ethanol and ethyl acetate extracts documented comparatively good activity against the studied microbial strains. The methanol extract of leaves of T. procumbens showed combatively better antioxidant potential. The tested plant leaf extract showed high activity against human lung cancer cells than breast cancer cell lines. 250 µg/ml plants extract showed 84 ± 2.8% toxicity against human lung cancer cells.

Prevalence and molecular characteristics of sequence type 131 clone among clinical uropathogenic Escherichia coli isolates in Riyadh, Saudi Arabia.

BACKGROUND: The antimicrobial resistance of extraintestinal pathogenic Escherichia coli (ExPEC) has progressively been reported worldwide. This resistance has been ascribed to global dissemination of a single E. coli clone, namely E. coli sequence type 131 (E. coli ST131). The main goal of this study is to determine the prevalence and molecular traits of ST131 and its subclones among E. coli clinical urine isolates in Riyadh, Saudi Arabia. METHODS: Sixty E. coli urine isolates, of different extended spectrum ß-lactamase (ESBL) carriage, were involved in this study. Molecular characterization was carried out to determine the ST131 status, phylogenetic groups and virulence carriage of these isolates. ST131 isolates were further tested to evaluate the prevalence of different phylogenetic groups, subclones and virulence carriage. RESULTS: Group B2 was the most common phylogroup from which E. coli isolates derived. Overall, 37 of 60 (61.7%) isolates belonged to ST131 clones. Of these, 19 (31.7%) isolates were from the H30 subclone, including 10 (16.7%) H30 non-Rx and 9 (15%) H30Rx. The remaining 18 (30%) ST131 isolates belonged to other non H30 subclones. H30 subclone was significantly higher in the virulence carriage in comparison to non H30 ST131 subclones. CONCLUSION: This study reported the prevalence and traits of clinical E. coli ST131 main subclones in Saudi Arabia. It also demonstrated the high prevalence of E. coli ST131 locally, and found different virulence genotypes and antimicrobial resistance phenotypes among ST131 subclones. In the future, preforming whole genome sequence-based studies on ST131 and its subclones is crucial to elucidate factors that drive the success of these organisms.

Purification and kinetics of the PHB depolymerase of Microbacterium paraoxydans RZS6 isolated from a dumping yard.

Poly-ß-hydroxybutyrate (PHB) depolymerase is known to decompose PHB, biodegradable polymers and therefore has great commercial significance in the bioplastic sector. However, reports on PHB depolymerases from isolates obtained from plastic-contaminated sites that reflect the potential of the source organism is scarce. In this study, we evaluated the production of extracellular PHB depolymerase from Microbacterium paraoxydans RZS6 isolated from the plastic-contaminated site in the municipal area of Shahada, Maharashtra, India, for the first time. The isolate was identified using 16S rRNA gene sequencing, gas chromatographic analysis of fatty acid methyl esters (GC-FAME), and BIOLOG method. Ithydrolyzed PHB on minimal salt medium (MSM) containing PHB as the only source of carbon. The isolate produced PHB depolymerase at 45°C during 48 h of incubation. The enzyme was purified most efficiently using octyl-sepharose CL-4B column, with the highest purification yield of 6.675 Umg-1mL-1. The activity of the enzyme was enhanced in the presence of Ca2+ and Mg2+ ions but inhibited by Fe2+ (1 mM) ions and mercaptoethanol (1000 rpm). the nzyme kinetic analysis revealed that the enzyme was a metalloenzyme; requiring Mg2+ ions, that showed optimum enzyme activity at 30°C (mesophilic) and under neutrophilic (pH 7) conditions. Scale-up from the shake-flask level to a laboratory-scale bioreactor further enhanced the enzyme yield by 0.809 UmL-1. The molecular weight of the enzyme (40 kDa), as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, closely resembled the PHB depolymerase of Aureobacterium saperdae. Our findings highlighted the applicability of M. paraoxydans as a producer of extracellular PHB depolymerase having potential of degrading PHB under diverse conditions.

Screening & analysis of anionic peptides from Foeniculum vulgare Mill by mass spectroscopy.

Fennel (Foeniculum vulgare Mill.) member from the family Umbelliferae (Apiaceae) and has been used in Saudi Arabia as an medicine as of the from the tradition. Our previous work with seed extracts of this plant generated DEAE-ion exchange purified proteins that exhibited antibacterial properties. The current study moves this work forward by using 2-D gel separation and MALDI TOF/TOF to identify proteins in this active extract. Fourteen protein spots were excised, digested, and identified. Several putative functions were identified, including: a copper-trans locating ATPase PAA1 chloroplastic-like isoform X1; a cytosolic enolase; a putative pentatricopeptide repeat-containing protein; an NADP-requiring isocitrate dehydrogenase; two proteins annotated as being encoded downstream from Son-like proteins; three probable nuclear proteins 5-1; and four predicted/ unidentified proteins. Future efforts will further characterize their relevant antimicrobial properties with the aim of cloning and high throughput synthesis of the antimicrobial element(s).

Prevalence of Multidrug Resistance and Extended-Spectrum ß-Lactamase Carriage of Clinical Uropathogenic Escherichia coli Isolates in Riyadh, Saudi Arabia.

The prevalence of extended-spectrum ß-lactamase-producing Escherichia coli (ESBL-producing E. coli) has recently increased worldwide. This study aims at determining the antimicrobial susceptibility patterns of a collection of clinical E. coli urine isolates and evaluating the ESBL carriage of these isolates at phenotypic and genotypic levels. A total of 100 E. coli urine isolates were collected at a tertiary healthcare centre in Riyadh from January 2018 to March 2018. Antimicrobial susceptibility testing was carried out for all isolates. ESBL production was characterized at phenotypic and genotypic levels using double-disc synergy test and polymerase chain reaction, respectively. Detection of different ESBL variants was performed using DNA sequencing. Of 100 E. coli isolates, 67 were associated with multidrug resistance (MDR) phenotype. All isolates showed variable resistance levels to all antibiotics used here expect to imipenem, where they were all imipenem-sensitive. 33 out of 100 E. coli isolates were positive for ESBLs by phenotypic and genotypic methods. Among all ESBL-positive E. coli isolates, the CTX-M was the most prevalent ESBL type (31/33 isolates; 93.94%). CTX-M-15 variant was detected in all isolates associated with CTX-M carriage. Multiple ESBL gene carriage was detected in 15/33 isolates (45.45%), where 11 (33.33%) isolates produced two different ESBL types while 4 isolates (12.12%) associated with carrying three different ESBL types. Our study documented the high antimicrobial resistance of ESBL-producing E. coli to many front-line antibiotics currently used to treat UTI patients, and this implies the need to continuously revise the local guidelines used for optimal empirical therapy for UTI patients. It also showed the high prevalence of ESBL carriage in E. coli urine isolates, with CTX-M-15 being the most predominant CTX-M variant.

Comparative genome analysis identifies few traits unique to the Escherichia coli ST131 H30Rx clade and extensive mosaicism at the capsule locus.

BACKGROUND: E.coli ST131 is a globally disseminated clone of multi-drug resistant E. coli responsible for that vast majority of global extra-intestinal E. coli infections. Recent global genomic epidemiological studies have highlighted the highly clonal nature of this group of bacteria, however there appears to be inconsistency in some phenotypes associated with the clone, in particular capsule types as determined by K-antigen testing both biochemically and by PCR. RESULTS: We performed improved quality assemblies on ten ST131 genomes previously sequenced by our group and compared them to a new reference genome sequence JJ1886 to identify the capsule loci across the drug-resistant clone H30Rx. Our data shows considerable genetic diversity within the capsule locus of H30Rx clone strains which is mirrored by classical K antigen testing. The varying capsule locus types appear to be randomly distributed across the H30Rx phylogeny suggesting multiple recombination events at this locus, but that this capsule heterogeneity has little to no effect on virulence associated phenotypes in vitro. CONCLUSIONS: Our data provides a framework for determining the capsular genetics of E. coli ST131 and further beyond to ExPEC strains, and highlights how capsular mosaicism may be an important strategy in becoming a successful globally disseminated human pathogen.

Phenotypic microarrays suggest Escherichia coli ST131 is not a metabolically distinct lineage of extra-intestinal pathogenic E. coli.

Extraintestinal pathogenic E. coli (ExPEC) are the major aetiological agent of urinary tract infections (UTIs) in humans. The emergence of the CTX-M producing clone E. coli ST131 represents a major challenge to public health worldwide. A recent study on the metabolic potential of E. coli isolates demonstrated an association between the E. coli ST131 clone and enhanced utilisation of a panel of metabolic substrates. The studies presented here investigated the metabolic potential of ST131 and other major ExPEC ST isolates using 120 API test reagents and found that ST131 isolates demonstrated a lower metabolic activity for 5 of 120 biochemical tests in comparison to non-ST131 ExPEC isolates. Furthermore, comparative phenotypic microarray analysis showed a lack of specific metabolic profile for ST131 isolates countering the suggestion that these bacteria are metabolically fitter and therefore more successful human pathogens.