The Antibacterial Activity of Rhazya stricta Extracts against Klebsiella pneumoniae Isolated from Some Soil Invertebrates at High Altitudes.

Klebsiella is a common dangerous pathogen for humans and animals and is widely present in the digestive system. The genus Klebsiella is ubiquitous, as it is endemic to surface water, soil, and sewage. In this study, 70 samples were obtained from soil-dwelling invertebrates from September 2021 to March 2022 from Taif and Shafa in different altitudinal regions of Saudi Arabia. Fifteen of these samples were identified as Klebsiella spp. The Klebsiella isolates were genetically identified as Klebsiella pneumoniae using rDNA sequencing. The antimicrobial susceptibility of the Klebsiella isolates was determined. Amplification of virulence genes was performed using PCR. In this study, 16S rDNA sequencing showed a similarity from 98% to 100% with related K. pneumonia from the NCBI database, and the sequences were deposited in the NCBI GenBank under accession numbers ON077036 to ON077050. The growth inhibition properties of ethanolic and methanolic extracts of the medicinal plant Rhazya stricta's leaves against K. pneumoniae strains using the minimum inhibitory concentration (MIC) method and disc diffusion were evaluated. In addition, the biofilm inhibitory potential of these extracts was investigated using crystal violet. HPLC analysis identified 19 components divided into 6 flavonoids, 11 phenolic acids, stilbene (resveratrol), and quinol, and revealed variations in the number of components and their quantities between extracts. Both extracts demonstrated interesting antibacterial properties against K. pneumoniae isolates. The 2 extracts also showed strong biofilm inhibitory activities, with percentages of inhibition extending from 81.5% to 98.7% and from 35.1% to 85.8% for the ethanolic and methanolic extracts, respectively. Rhazya stricta leaf extract revealed powerful antibacterial and antibiofilm activities against K. pneumoniae isolates and could be a good candidate for the treatment or prevention of K. pneumonia-related infections.

GC-MS analysis, phytochemical composition of Hertia cheirifolia L. essential oil with pharmacological assessments: antioxidant, antibacterial, and antifungal activities.

The genus Hertia, which belongs to the Asteraceae family, is a flowering genus with 12 species found in Africa, North and South. Among the species present in Algeria, Hertia cheirifolia L. is distributed in the eastern regions of Algeria. The aim of this study is to evaluate its phytochemical composition with following pharmacological assessments: the antioxidant, antibacterial, and antifungal activities of Hertia cheirifolia L. essential oil (EO). GC-MS analysis was used to analyze the chemical constituents of H. cheirifolia essential oil. The antioxidant capacity was assessed using DPPH, FRAP, and H2O2 tests. The EO was also tested for its ability to inhibit six strains of microorganisms, including two Gram (+) and four Gram (-) strains. The antifungal activity was tested by analyzing the effect of the EO on the mycelial growth of Fusarium oxysporum f.sp. lycopersici (FOL) fungi. Results showed that primary volatile components were α-pinene (32.59%), 2-(1-cyclopent-1-enyl-1-methylethyl) cyclopentanone (14.62%), (-)-germacrene D (11.37%), and bakkenolide A (9.57%). H. cheirifolia EO showed inhibitory effects against DPPH, H2O2, and FRAP (IC50 = 0.34 ± 0.1, 0.053 ± 0.1, and 0.047 ± 0.01 mg mL-1, respectively). The EO also exhibited moderate antibacterial effects against Staphylococcus aureus ATCC 25923 (S. aureus), Streptococcus pneumoniae ATCC 49619 (S. pneumoniae), and Enterobacter aerogenes ATCC 13048 (E. aerogenes), as well as significant antioxidant potential and varied antifungal activity based on dosage and fungal strain. To our knowledge, no previous research has examined the antifungal capacity of H. cheirifolia oil and oil-mycelial development of the FOL relationship. To fully explore the benefits of H. cheirifolia EO, more in vivo research is necessary, along with more testing on other bacterial and fungal strains.

Efficiency of Bacillus thuringiensis and Bacillus cereus against Rhynchophorus ferrugineus.

The Red Palm Weevil (Rhynchophorus ferrugineus (Oliv.) (Coleoptera, Dryophthoridae) is a well-known palm tree pest that has caused enormous economic damage all over the globe. Insecticides are still the primary method of controlling this pest at this period. However, field populations of RPW have been shown to be resistant to pesticides. Using Bacillus spp. might be one of the options for controlling R. ferruginous. In this study, 23 species of Bacillus spp. were isolated from the rhizosphere of date palm trees in Al Ahsa Oasis, Saudi Arabia. The isolates were identified using 16S rRNA gene sequencing. R. ferrugineus larvae and adults were tested on sugarcane pieces that were treated with the B. thuringiensis strain PDC-AHSAA1 and B. cereus strains (PDC-AHSAA2, PDC-AHSA3 and PDC-AHSA4). The LC50 values for larvae and adults were quite low when they were compared with those of the other isolated strains. The B. thuringiensis strain PDC-AHSAA1 was more effective against both the larvae and adults. The determined LC50 values for B. thuringiensis ranged from 4.19 × 107-3.78 × 109. After 21 days, the data on larval mortality and body weight were evaluated. The surviving larvae that were treated with the bacterial isolates did not acquire a substantial weight. For the RPW larvae and adults, the mortality and corrected mortality death rates were increased by increasing the concentration of B. thuringiensis. In conclusion, Bacillus-treated diets negatively influenced the growth and development of the RPW. This research reported on the interaction between the RPW and the rhizosphere Bacillus spp. and highlighted the tremendous potential for the development of microbial resource-based control strategies for this pest.

Harnessing the Potential of Bacillus altitudinis MT422188 for Copper Bioremediation.

The contamination of heavy metals is a cause of environmental concern across the globe, as their increasing levels can pose a significant risk to our natural ecosystems and public health. The present study was aimed to evaluate the ability of a copper (Cu)-resistant bacterium, characterized as Bacillus altitudinis MT422188, to remove Cu from contaminated industrial wastewater. Optimum growth was observed at 37°C, pH 7, and 1 mm phosphate, respectively. Effective concentration 50 (EC50), minimum inhibitory concentration (MIC), and cross-heavy metal resistance pattern were observed at 5.56 mm, 20 mm, and Ni > Zn > Cr > Pb > Ag > Hg, respectively. Biosorption of Cu by live and dead bacterial cells in its presence and inhibitors 1 and 2 (DNP and DCCD) was suggestive of an ATP-independent efflux system. B. altitudinis MT422188 was also able to remove 73 mg/l and 82 mg/l of Cu at 4th and 8th day intervals from wastewater, respectively. The presence of Cu resulted in increased GR (0.004 ± 0.002 Ug-1FW), SOD (0.160 ± 0.005 Ug-1FW), and POX (0.061 ± 0.004 Ug-1FW) activity. Positive motility (swimming, swarming, twitching) and chemotactic behavior demonstrated Cu as a chemoattractant for the cells. Metallothionein (MT) expression in the presence of Cu was also observed by SDS-PAGE. Adsorption isotherm and pseudo-kinetic-order studies suggested Cu biosorption to follow Freundlich isotherm as well as second-order kinetic model, respectively. Thermodynamic parameters such as Gibbs free energy (∆G°), change in enthalpy (∆H° = 10.431 kJ/mol), and entropy (∆S° = 0.0006 kJ/mol/K) depicted the biosorption process to a feasible, endothermic reaction. Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Energy-Dispersive X-Ray Spectroscopy (EDX) analyses revealed the physiochemical and morphological changes in the bacterial cell after biosorption, indicating interaction of Cu ions with its functional groups. Therefore, these features suggest the potentially effective role of B. altitudinis MT422188 in Cu bioremediation.

Potential Therapeutic Candidates against Chlamydia pneumonia Discovered and Developed In Silico Using Core Proteomics and Molecular Docking and Simulation-Based Approaches.

Chlamydia pneumonia, a species of the family Chlamydiacea, is a leading cause of pneumonia. Failure to eradicate C. pneumoniae can lead to chronic infection, which is why it is also considered responsible for chronic inflammatory disorders such as asthma, arthritis, etc. There is an urgent need to tackle the major concerns arising due to persistent infections caused by C. pneumoniae as no FDA-approved drug is available against this chronic infection. In the present study, an approach named subtractive proteomics was employed to the core proteomes of five strains of C. pneumonia using various bioinformatic tools, servers, and software. However, 958 non-redundant proteins were predicted from the 4754 core proteins of the core proteome. BLASTp was used to analyze the non-redundant genes against the proteome of humans, and the number of potential genes was reduced to 681. Furthermore, based on subcellular localization prediction, 313 proteins with cytoplasmic localization were selected for metabolic pathway analysis. Upon subsequent analysis, only three cytoplasmic proteins, namely 30S ribosomal protein S4, 4-hydroxybenzoate decarboxylase subunit C, and oligopeptide binding protein, were identified, which have the potential to be novel drug target candidates. The Swiss Model server was used to predict the target proteins' three-dimensional (3D) structure. The molecular docking technique was employed using MOE software for the virtual screening of a library of 15,000 phytochemicals against the interacting residues of the target proteins. Molecular docking experiments were also evaluated using molecular dynamics simulations and the widely used MM-GBSA and MM-PBSA binding free energy techniques. The findings revealed a promising candidate as a novel target against C. pneumonia infections.