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  1. Al-Kafaween MA, Nagi Al-Jamal HA
    Iran J Microbiol, 2022 Apr;14(2):238-251.
    PMID: 35765547 DOI: 10.18502/ijm.v14i2.9193
    BACKGROUND AND OBJECTIVES: Honey has excellent antibacterial properties against various microorganisms of several different species. To date, there is no comparative evaluation of the antibacterial activity of Jarrah honey (JH), Kelulut Madu honey (KMH), Gelam honey (GH), and Acacia honey (AH) with that of Manuka honey (MH). The purpose of this study was to conduct such study and to compare the antibacterial activity of JH, KMH, GH, and AH with that of MH against Pseudomonas aeruginosa and Streptococcus pyogenes.

    MATERIALS AND METHODS: Activity was assessed using broth microdilution, time kill viability, microtiter plate, scanning electron microscope (SEM) and Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR).

    RESULTS: The susceptibility tests revealed promising antibacterial activities of all honeys against both bacteria. The MICs of JH, KMH, GH, and AH ranged from 20% to 25% compared to MH (12.5%) against both bacteria. The MBCs of JH, KMH, GH, and AH ranged from 20% to 50% compared to MH (20%) against both bacteria. Treatment of both bacteria with 2× MIC (Minimum inhibitory concentration) of MH, JH, KMH, GH, and AH for 9 hours resulted in reduction in colony-forming unit (CFU/ml). SEM images showed that the morphological changes, cell destruction, cell lysis and biofilm disruption in both bacteria after exposure to all honeys. RT-qPCR analysis revealed that the expression of all genes in both bacteria were downregulated following treatment with all honeys. Among the all-tested honeys, MH showed the highest total antibacterial and antivirulence activities.

    CONCLUSION: Our results indicate that all honeys activity included inhibition of both bacteria due to a decrease in expression of essential genes associated with both bacteria, suggesting that all honeys could potentially be used as an alternative therapeutic agent against certain microorganisms particularly against P. aeruginosa and S. pyogenes.

  2. Al-Kafaween MA, Al-Groom RM, Hilmi ABM
    Iran J Microbiol, 2023 Feb;15(1):89-101.
    PMID: 37069905 DOI: 10.18502/ijm.v15i1.11923
    BACKGROUND AND OBJECTIVES: Honey is one of the oldest traditional remedies that has been widely utilized to cure a variety of human ailments. The objective of this research was to test and compare the antibacterial activity of Sidr honey (SH) and Tualang honey (TH) to that of Manuka honey (MH) against Staphylococcus aureus.

    MATERIALS AND METHODS: The antibacterial activity of MH, SH and TH against S. aureus was investigated by agar well diffusion, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), time-kill curve, microtiter plate and RT-qPCR analysis.

    RESULTS: Agar inhibition assay showed that MH possess highest total antibacterial activity against S. aureus with an inhibition zone 25.1 mm compared with that of SH (22.2 mm) and TH (21.3 mm). The findings showed that when compared to SH and TH (MIC: 25% and MBC: 50%), MH honey had the lowest MIC (12.5%) and MBC (25%). After S. aureus was exposed to MH, SH, and TH, there was a decrease in colony-forming unit as seen by the time-kill curve. The lowest concentration 20% of MH, SH and TH was significantly found to inhibit S. aureus biofilm. The RT-qPCR results revealed that all the selected genes in S. aureus were downregulated in gene expression following exposure to each of the tested honeys. Comparing the total antibacterial, antibiofilm, and antivirulence activities of all the tested honeys, MH demonstrated the greatest levels of these properties.

    CONCLUSION: According to this study, various types of each evaluated honey have the capacity to effectively suppress and modify the virulence of S. aureus via a variety of molecular targets.

  3. Abu-Sini M, Al-Kafaween MA, Al-Groom RM, Hilmi ABM
    AIMS Microbiol, 2023;9(2):313-331.
    PMID: 37091817 DOI: 10.3934/microbiol.2023017
    P. aeruginosa is an opportunistic pathogen that is commonly found in nosocomial infections. The purpose of this study was to investigate the effects of seven antibiotics on P. aeruginosa planktonic growth, biofilm formation, and the expression of virulence factors. These antibiotics included Ciprofloxacin (CP), Amikacin (AMK), Vancomycin (VAN), Tetracycline (TET), Gentamicin (GEN), Erythromycin (Ery), and Clindamycin (CLI). Antibiotic susceptibility testing, Minimum Bactericidal Concentration (MBC), Minimum Inhibitory Concentration (MIC), growth curve, time-kill curve, biofilm inhibition and reduction assay, and RT-qPCR were used to assess the effects of these antibiotics on P. aeruginosa planktonic and biofilm. The clear zones of inhibition against P. aeruginosa for the CP, AMK, VAN, TET, GEN, Ery, and CLI were 26 mm, 20 mm, 21 mm, 22 mm, 20 mm, 25 mm and 23 mm, respectively. The MIC values for CP, AMK, VAN, TET, GEN, Ery and CLI against P. aeruginosa ranged from 0.25 to 1 µg/mL while the MBC values ranged from 1 and 0.5 to 2 µg/mL respectively. The growth, total viable counts (TVCs), bacterial adhesion and biofilm formation of P. aeruginosa were reduced after exposure to all the tested antibiotics in a dose-dependent manner. The RT-qPCR analysis showed that all the tested antibiotics share a similar overall pattern of gene expression, with a trend toward reduced expression of the virulence genes of interest (lasR, lasI, fleN, fleQ and fleR, oprB and oprC) in P. aeruginosa. The results indicate that all of the tested antibiotics possess antimicrobial and anti-biofilm activities, and that they may be multiple inhibitors and moderators of P. aeruginosa virulence via a variety of molecular targets. This deduction requires to be investigated in vivo.
  4. Al-Kafaween MA, Al-Jamal HAN, Hilmi ABM, Elsahoryi NA, Jaffar N, Zahri MK
    Iran J Microbiol, 2020 Dec;12(6):565-576.
    PMID: 33613911 DOI: 10.18502/ijm.v12i6.5031
    BACKGROUND AND OBJECTIVES: Tualang honey (TH) is a Malaysian multifloral jungle honey. In recent years, there has been a marked increase in the number of studies published in medical databases regarding its potential health benefits. The study aimed to investigate the effect of TH against Pseudomonas aeruginosa and Streptococcus pyogenes.

    MATERIALS AND METHODS: The effect of TH on both bacteria was investigated using MIC, MBC, growth curve, time-kill curve, scanning electron microscopy (SEM) and RT-qPCR.

    RESULTS: The MIC of TH against P. aeruginosa and S. pyogenes was 18.5% (w/v) and 13% (w/v) respectively and MBC was 25% (w/v) for both bacteria. Spectrophotometric readings of at least 90% inhibition yielded MIC90 values of TH, 18.5% (w/v) and 15% (w/v) for P. aeruginosa and S. pyogenes respectively. A time-kill curve demonstrated a bactericidal with a 4-log reduction estimated within 8 hours. Using SEM, loss of structural integrity and marked changes in cell shape were observed. RT-qPCR analysis showed that TH reduced the pattern of gene expression in both bacteria, with a trend toward reduced expression of the virulence genes of interest.

    CONCLUSION: This study suggests that TH could potentially be used as an alternative therapeutic agent for microbial infection particularly against these two organisms.

  5. Khleifat KM, Al-Tawarah NM, Al-Kafaween MA, Al-Ksasbeh W, Qaralleh H, Alqaraleh M, et al.
    Curr Alzheimer Res, 2023;20(3):190-201.
    PMID: 37317907 DOI: 10.2174/1567205020666230614143027
    BACKGROUND/OBJECTIVE: Alzheimer's disease (AD) is mainly characterized by amnesia that affects millions of people worldwide. This study aims to explore the effectiveness capacities of bee venom (BV) for the enhancement of the memory process in a rat model with amnesia-like AD.

    METHODS: The study protocol contains two successive phases, nootropic and therapeutic, in which two BV doses (D1; 0.25 and D2: 0.5 mg/kg i.p.) were used. In the nootropic phase, treatment groups were compared statistically with a normal group. Meanwhile, in the therapeutic phase, BV was administered to scopolamine (1mg/kg) to induce amnesia-like AD in a rat model in which therapeutic groups were compared with a positive group (donepezil; 1mg/kg i.p.). Behavioral analysis was performed after each phase by Working Memory (WM) and Long-Term Memory (LTM) assessments using radial arm maze (RAM) and passive avoidance tests (PAT). Neurogenic factors; Brain-derived neurotrophic factor (BDNF), and Doublecortin (DCX) were measured in plasma using ELISA and Immunohistochemistry analysis of hippocampal tissues, respectively.

    RESULTS: During the nootropic phase, treatment groups demonstrated a significant (P < 0.05) reduction in RAM latency times, spatial WM errors, and spatial reference errors compared with the normal group. In addition, the PA test revealed a significant (P < 0.05) enhancement of LTM after 72 hours in both treatment groups; D1 and D2. In the therapeutic phase, treatment groups reflected a significant (P < 0.05) potent enhancement in the memory process compared with the positive group; less spatial WM errors, spatial reference errors, and latency time during the RAM test, and more latency time after 72 hours in the light room. Moreover, results presented a marked increase in the plasma level of BDNF, as well as increased hippocampal DCX-positive data in the sub-granular zone within the D1 and D2 groups compared with the negative group (P < 0.05) in a dose-dependent manner.

    CONCLUSION: This study revealed that injecting BV enhances and increases the performance of both WM and LTM. Conclusively, BV has a potential nootropic and therapeutic activity that enhances hippocampal growth and plasticity, which in turn improves WM and LTM. Given that this research was conducted using scopolamine-induced amnesia-like AD in rats, it suggests that BV has a potential therapeutic activity for the enhancement of memory in AD patients in a dose-dependent manner but further investigations are needed.

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