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Published on: September 3, 2016
Inhibition of Staphylococcus aureus LC554891 by Moringa oleifera Seed Extract either Singly or in Combination with
Gamal Enan1, Abdul-Raouf Al-Mohammadi2, Samir Mahgoub3
1Department of Botany and Microbiology, Faculty of Science, Zagazig University, Zagazig 44519, Egypt.
Abstract:
Bacterial outbreaks caused by Staphylococcus aureus (S. aureus) are interesting due to the existence of multidrug resistant (MDR) isolates. Therefore, there is a need to develop novel ways to control such MDR S. aureus. In this study, some natural agents such as honey bee (HB), extracts of either Moringa oleifera seeds (MSE), or leaves (MLE) and essential oils of garlic, clove, and moringa were studied for their inhibitory activity against this S. aureus pathogen. About 100 food samples including beef luncheon (n = 25), potato chips (n = 50), and corn flakes (n = 25) were investigated for possible pollution with the S. aureus bacteria. The isolated bacteria suspected to belong S. aureus that grew well onto Baird-Parker agar (Oxoid) and shiny halo zones and positive coagulase reaction were selected and identified by API-Kits; all of them that were approved belong to S. aureus (18 strains). The sensitivity of the obtained 18 S. aureus bacterial strains to 12 antibiotics were evaluated; all of them were resistant to ofloxacin; however, other antibiotics tested showed variable results. Interestingly, the S. aureus No. B3 isolated from beef luncheon was resistant to10 antibiotics out of 12 ones tested. Multiple antibiotic resistance index (MAR) of this S. aureus strain was about 83.3%. Therefore, its identification was confirmed by sequencing of a 16S rRNA gene which approved a successful biochemical identification carried out by API Kits and such strain was designated S. aureus LC 554891. The genome of such strain appeared to contain mecA gene encoding methicillin resistance; it was found to contain hla, hlb, tsst-1, and finbA that encode α-blood hemolysis, β-blood hemolysis, toxic shock syndrome gene, and fibrinogen-binding protein gene, respectively. In addition, the virulence factors viz. sea; seb; sec encoding enterotoxins were detected in the DNA extracted from S. aureus B3 strain. Aqueous extract of Moringa oleifera seeds (MSE) showed inhibitory activity against S. aureus LC 554891 better than that obtained by tetracycline, essential oils or HB. Minimum inhibitory concentration (MIC) of MSE was 20µg/mL. Instrumental analysis of MSE showed 14 bioactive chemical compounds. Combinations of both MSE and tetracycline showed distinctive inhibitory activity against S. aureus LC 554891 than that obtained by either tetracycline or MSE singly.
Insights
This study investigated natural agents against multidrug-resistant Staphylococcus aureus (S. aureus). Moringa oleifera seed extract (MSE) demonstrated significant inhibitory activity, outperforming antibiotics and other natural agents against a highly resistant S. aureus strain.
Area of Science:
- Microbiology and Infectious Diseases
- Natural Product Chemistry
- Food Safety
Background:
- Multidrug-resistant (MDR) Staphylococcus aureus (S. aureus) poses a significant threat due to limited treatment options.
- The emergence of MDR S. aureus in food products necessitates the development of novel control strategies.
- Natural agents offer potential alternatives for combating antibiotic-resistant bacterial pathogens.
Purpose of the Study:
- To evaluate the antimicrobial activity of natural agents against MDR S. aureus isolated from food samples.
- To identify and characterize a highly resistant S. aureus strain and assess its virulence factors.
- To determine the efficacy of Moringa oleifera seed extract (MSE) as a potential therapeutic agent.
Main Methods:
- Isolation and identification of S. aureus from 100 food samples using selective media and API Kits.
- Antibiotic susceptibility testing of isolated S. aureus strains, including MAR index calculation.
- Genomic analysis (16S rRNA sequencing, PCR for virulence genes like mecA, hla, hlb, tsst-1, sea, seb, sec) and instrumental analysis of MSE.
- Determination of Minimum Inhibitory Concentration (MIC) for MSE and evaluation of combined effects with tetracycline.
Main Results:
- Eighteen S. aureus strains were isolated, with one strain (S. aureus LC 554891) exhibiting resistance to 10 out of 12 antibiotics (MAR index 83.3%).
- This MDR strain possessed virulence genes including mecA (methicillin resistance), hemolytic toxins, toxic shock syndrome gene, and enterotoxin genes (sea, seb, sec).
- Aqueous MSE showed superior inhibitory activity against S. aureus LC 554891 (MIC 20µg/mL) compared to tetracycline, essential oils, and honey bee extract.
- Combinations of MSE and tetracycline exhibited enhanced inhibitory effects.
Conclusions:
- Moringa oleifera seed extract (MSE) is a promising natural agent for controlling MDR S. aureus.
- The identified MDR S. aureus strain possesses significant virulence factors, highlighting the need for effective control measures.
- Further research into MSE's bioactive compounds and synergistic effects with antibiotics could lead to novel therapeutic strategies.
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