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Synergistic evaluation of Moringa oleifera extract and ß-lactam antibiotic to restore the susceptibility of methicillin-resistant Staphylococcus aureus

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Abstract

Introduction

The antibiotic resistance has become a major threat to global health. The combinatorial use of two or more compounds to develop a new formulation may overcome the emerging cases of drug resistance. Moringa oleifera has been utilized as a strong nutritional, immunomodulator and therapeutic agent for decades. In this study, different parts of Moringa oleifera were screened for bioactive compounds that can act as a resistance modifying agent for multi-drug resistant organisms (MDROs).

Methodology

Initially, the combined effect of stem bark extracts and ampicillin was calculated by checkerboard assay. Active compounds of effective extract were assessed by High Performance Liquid Chromatography (HPLC). Minimal Inhibitory Concentration (MIC) and Fractional Inhibitory Concentration Index (FICI) were calculated to evaluate the synergistic behavior of stem bark extract with ampicillin. To study the blocking of resistance pathways of Methicillin-Resistant Staphylococcus aureus (MRSA) western blot was performed.

Results

The results revealed that stem bark has significant anti-MRSA activity. The methanolic extract of stem bark in combination with ampicillin showed the highest synergistic effect (FICI value ≤ 0.237) against MRSA. Killing kinetics and membrane potential of ampicillin alone and in combination revealed an increase in the inhibitory potential of ampicillin against MRSA. Decolourization in iodometric assay confirmed the inhibition of β-lactamase, western blot results confirmed the blocking of penicillin-binding protein (PBP2a) expression with the restoration of MRSA sensitivity against β-lactams.

Conclusion

It can be concluded that methanolic extract of Moringa oleifera stem bark has bioactive compounds and can be used as an adjuvant with antibiotics to modify the resistance of MDROs.

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Abbreviations

MRSA:

Methicillin-resistant staphylococcus aureus

MDR:

Multi-drug resistance

MDROs:

Multidrug resistant organisms

TPC:

Total phenolic content

TFC:

Total flavonoid content

MICs:

Minimal inhibitory concentrations

FICI:

Fractional inhibitory concentration index

DMSO:

Dimethyl sulfoxide

PAGE:

Polyacrylamide gel electrophoresis

PBP2a:

Penicillin binding protein 2a

HPLC:

High-performance liquid chromatography

PBS:

Phosphate buffered saline

BCIP:

5-Bromo-4-chloro-3-indolylphosphate

CFU:

Colony forming unit

SDS-PAGE:

Sodium dodecyl sulfate

NBT:

Nitro blue tetrazolium

ALP:

Alkaline phosphatase

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Acknowledgements

The authors gratefully acknowledge the KIBGE, University of Karachi, Karachi, Pakistan for providing all the laboratory facilities and financial assistance. We would like to thank Editage (www.editage.com) for English language editing.

Funding

There was no funding was available to support this project.

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Authors and Affiliations

  1. The Karachi Institute of Biotechnology & Genetic Engineering (KIBGE), University of Karachi, Karachi, 75270, Pakistan

    Soma Vankwani, Asma Ansari, Abid Azhar & Saddia Galani

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  1. Soma Vankwani
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  2. Asma Ansari
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  3. Abid Azhar
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  4. Saddia Galani
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Contributions

This study was initially conceived and designed by SV and SG. Methodology and protocols were finalized by SV, SG, AA. While experiments handling and data collection was ended by SV. Whereas data analysis and interpretation by SG, AA, AAZ. For manuscript writing the initial draft was prepared by SV that was further reviewed and edited by SG, AA, AAZ.

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Correspondence to Saddia Galani.

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Vankwani, S., Ansari, A., Azhar, A. et al. Synergistic evaluation of Moringa oleifera extract and ß-lactam antibiotic to restore the susceptibility of methicillin-resistant Staphylococcus aureus. Mol Biol Rep 49, 421–432 (2022). https://doi.org/10.1007/s11033-021-06889-7

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