Browsing by Author "AbdulAzeez, Sayed"

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Entomopathogenic fungi and their biological control of Tetranychus urticae: Two-spotted spider mites
(2023) Khamis Al-Zahrani, Jawaher; Amira Alabdalall; Aly Osman, Mohamed; Aldakheel, Lena A.; Faisal AlAhmady, Nada; Aldakeel, Sumayh A.; J. Francis Borgio; Francis Borgio, J.; ElNaggar, Medhat A.; Alabdallah, Nadiyah M.; Almustafa, Mona M.
The two-spotted spider mite (TSSM) Tetranychus urticae, is regarded as one of the most dangerous pests responsible for great losses in most of agricultural crops. It is a persistent pest in Saudi Arabia, especially in greenhouses where T. urticae is primarily controlled by chemical pesticides. The main problem for the two-spotted spider mites is its high resistance to pesticides and high fertility rate. In the long term, chemical pesticides cause health problems and economic losses, so it was necessary to search for a safe alternative method for human health and the environment. One of these alternative methods was the selection of plant varieties resistant to the TSSM, in addition to biological control that includes mites or predatory insects and entomopathogenic fungi. The growth, reproduction, and life-table parameters of T. urticae were examined in a laboratory setting with a 16L:8D photoperiod at 28 ± 1 °C and 65 ± 5% RH, in the presence of three major members of Family: Solanaceae tomato, eggplant, and pepper. Pepper was shown to be less conducive to T. urticae growth and reproduction compared to eggplant and tomato. Tetranychus urticae proceeded through all five stages of its life cycle (egg, larva, protonymph, deutonymph, and adult) on tested solanaceous plants, and these plants significantly influenced its growth, reproduction, and Life-table parameters. Additionally, entomopathogenic fungi have been used against insects that have proven highly effective in controlling and reducing the density of two-spotted spider mites. Eight fungi were isolated from 80 insect and mite samples collected from several Saudi Arabia regions. Analysis of the 18S rRNA sequences revealed that the fungal strains identified as Beauveria bassiana, Fusarium sp. F. equiseti, F. oxysporium, Scopulariopsis brevicaulis1, S. brevicaulis2, Aspergillus sclerotiorum, and Penicillium citrinum. The ability of isolated fungi to secrete enzymes degrading the two-spotted spider mite cuticle, namely lipase, protease, and chitinase, were studied.
Genomic Landscape of Multidrug Resistance and Virulence in Enterococcus faecalis IRMC827A from a Long-Term Patient
(2023) Rahaf Khalid Al-Quwaie; AlJindan, Reem; Razan Aldahhan; Alquwaie, Rahaf; J. Francis Borgio; Reem AlJindan; AlEraky, Doaa M.; Noor Barak Almandil; Sayed AbdulAzeez; Doaa Mostafa AlEraky
We report on a highly virulent, multidrug-resistant strain of Enterococcus faecalis IRMC827A that was found colonizing a long-term male patient at a tertiary hospital in Khobar, Saudi Arabia. The E. faecalis IRMC827A strain carries several antimicrobial drug resistance genes and harbours mobile genetic elements such as Tn6009, which is an integrative conjugative element that can transfer resistance genes between bacteria and ISS1N via an insertion sequence. Whole-genome-sequencing-based antimicrobial susceptibility testing on strains from faecal samples revealed that the isolate E. faecalis IRMC827A is highly resistant to a variety of antibiotics, including tetracycline, doxycycline, minocycline, dalfopristin, virginiamycin, pristinamycin, chloramphenicol, streptomycin, clindamycin, lincomycin, trimethoprim, nalidixic acid and ciprofloxacin. The isolate IRMC827A carries several virulence factors that are significantly associated with adherence, biofilm formation, sortase-assembled pili, manganese uptake, antiphagocytosis, and spreading factor of multidrug resistance. The isolate also encompasses two mutations (G2576T and G2505A) in the 23S rRNA gene associated with linezolid resistance and three more mutations (gyrA p.S83Y, gyrA p.D759N and parC p.S80I) of the antimicrobial resistance phenotype. The findings through next-generation sequencing on the resistome, mobilome and virulome of the isolate in the study highlight the significance of monitoring multidrug-resistant E. faecalis colonization and infection in hospitalized patients. As multidrug-resistant E. faecalis is a serious pathogen, it is particularly difficult to treat and can cause fatal infections. It is important to have quick and accurate diagnostic tests for multidrug-resistant E. faecalis, to track the spread of multidrug-resistant E. faecalis in healthcare settings, and to improve targeted interventions to stop its spread. Further research is necessary to develop novel antibiotics and treatment strategies for multidrug-resistant E. faecalis infections.
Isolation, Screening, and Identification of Alkaline Protease-Producing Bacteria and Application of the Most Potent Enzyme from Bacillus sp. Mar64
(2023) Kotb, Essam; Essam Kotb; Alsayed, Mariam A.; Azzah Ibrahim Alghamdi; Alkhaldi, Eida; AbdulAzeez, Sayed; Sayed Abdul Azeez
In this study, thirty-seven alkaline protease-producing bacteria were recovered from different regions of Saudi Arabia. The proteolytic strain with the highest productivity was identified as Bacillus sp. Mar64. Maximum productivity of Mar64P alkaline protease was reached at 60 h, pH 9.0, and 45 °C using 1% tyrosine and 0.5% maltose as nitrogen and carbon supplies, respectively. Specific activity was intensified to 8.5-fold with a recovery of 12.4% and SDS—PAGE revealed one band at 28 kDa after enzyme purification. Mar64P was maximally active at 55 °C and pH 11.0 with thermal stability up to 70 °C and pH stability at 7.0–12.0 for 1 h. It was inhibited by EDTA and unaffected by PMSF, therefore tentatively classified as metalloprotease-type. Storage efficacy was effective for up to eight weeks and it was durable in presence of organic solvents (20%, v/v) such as acetonitrile, acetone, and isopropanol upto to 15 days. The enzyme was compatible with dry detergents at both low and high temperature, in addition, was successful in removing various stains such as blood, egg yolk, chocolate, tea, coffee, and sweat. Furthermore, it was successful in removing skin hairs and hydrolyzing gelatin of waste X-ray films. Collectively, due to these unique properties, Mar64P could be considered an environmentally friendly candidate in both detergent and leather industries.
Mining the nanotube-forming Bacillus amyloliquefaciens MR14M3 genome for determining anti-Candida auris and anti-Candida albicans potential by pathogenicity and comparative genomics analysis
(2023) Borgio, J. Francis; Alhujaily, Rahaf; Eman saleh Alhasani; Alabdullah, Maryam Jawad; AlHasani, Eman; Alothman, Wojod; Alaqeel, Rawan Khalid; Rahaf Alhujaily; Kaabi, Ayidah; Alhur, Norah F.; Akhtar, Sultan; Almandil"; Almofty, Sarah; Almofty"; AbdulAzeez, Sayed; "Sayed; AbdulAzeez"; Reem AlJindan
There is a global health concern associated with the emergence of the multidrug-resistant (MDR) fungus Candida auris, which has significant mortality rates. Finding innovative and distinctive anti-Candida compounds is essential for treating infections caused by MDR C. auris. A bacterial strain with anti-Candida activity was isolated and identified using 16 S rRNA gene sequencing. The whole genome was sequenced to identify biosynthesis-related gene clusters. The pathogenicity and cytotoxicity of the isolate were analyzed in Candida and HFF-1 cell lines, respectively. This study set out to show that whole-genome sequencing, cytotoxicity testing, and pathogenicity analysis combined with genome mining and comparative genomics can successfully identify biosynthesis-related gene clusters in native bacterial isolates that encode antifungal natural compounds active against Candida albicans and C. auris. The native isolate MR14M3 has the ability to inhibit C. auris (zone of inhibition 25 mm) and C. albicans (zone of inhibition 25 mm). The 16 S rRNA gene sequence of MR14M3 aligned with Bacillus amyloliquefaciens with similarity (100%). Bacillus amyloliquefaciens MR14M3 establishes bridges of intercellular nanotubes (L 258.56 ± 35.83 nm; W 25.32 ± 6.09 nm) connecting neighboring cells. Candida cell size was reduced significantly, and crushed phenotypes were observed upon treatment with the defused metabolites of B. amyloliquefaciens MR14M3. Furthermore, the pathogenicity of B. amyloliquefaciens MR14M3 on Candida cells was observed through cell membrane disruption and lysed yeast cells. The whole-genome alignment of the MR14M3 genome (3981,643 bp) using 100 genes confirmed its affiliation with Bacillus amyloliquefaciens. Genome mining analysis revealed that MR14M3-coded secondary metabolites are involved in the biosynthesis of polyketides (PKs) and nonribosomal peptide synthases (NRPSs), including 11 biosynthesis-related gene clusters with one hundred percent similarity. Highly conserved biosynthesis-related gene clusters with anti-C. albicans and anti-C. auris potentials and cytotoxic-free activity of B. amyloliquefaciens MR14M3 proposes the utilization of Bacillus amyloliquefaciens MR14M3 as a biofactory for an anti-Candida auris and anti-C. albicans compound synthesizer.
Therapeutic Intervention for Various Hospital Setting Strains of Biofilm Forming Candida auris with Multiple Drug Resistance Mutations Using Nanomaterial Ag-Silicalite-1 Zeolite
(2022) Aldossary, Hanan A.; Ebtesam Abdullah Al-Suhaimi; Jermy, B. Rabindran; AlJindan, Reem; Aldayel, Afra; AbdulAzeez, Sayed; Akhtar, Sultan; Khan, Firdos Alam; Borgio, J. Francis; Al-Suhaimi, Ebtesam Abdullah
Candida auris (C. auris), an emerging multidrug-resistant microorganism, with limited therapeutical options, is one of the leading causes of nosocomial infections. The current study includes 19 C. auris strains collected from King Fahd Hospital of the University and King Fahad Specialist Hospital in Dammam, identified by 18S rRNA gene and ITS region sequencing. Drug-resistance-associated mutations in ERG11, TAC1B and FUR1 genes were screened to gain insight into the pattern of drug resistance. Molecular identification was successfully achieved using 18S rRNA gene and ITS region and 5 drug-resistance-associated missense variants identified in the ERG11 (F132Y and K143R) and TAC1B (H608Y, P611S and A640V) genes of C. auris strains, grouped into 3 clades. The prophylactic and therapeutic application of hydrothermally synthesized Ag-silicalite-1 (Si/Ag ratio 25) nanomaterial was tested against the 3 clades of clinical C. auris strains. 4wt%Ag/TiZSM-5 prepared using conventional impregnation technique was used for comparative study, and nano formulations were characterized using different techniques. The antibiofilm activity of nanomaterials was tested by cell kill assay, scanning electron microscopy (SEM) and light microscopy. Across all the clades of C. auris strains, 4 wt%Ag/TiZSM-5 and Ag-silicalite-1 demonstrated a significant (p = 1.1102 × 10−16) inhibitory effect on the biofilm’s survival rate: the lowest inhibition value was (10%) with Ag-silicalite-1 at 24 and 48 h incubation. A profound change in morphogenesis in addition to the reduction in the number of C. auris cells was shown by SEM and light microscopy. The presence of a high surface area and the uniform dispersion of nanosized Ag species displays enhanced anti-Candida activity, and therefore it has great potential against the emerging multidrug-resistant C. auris.