Antimicrobial and Anti-Biofilm Activities of Citrus sinensis and Moringa oleifera Against the Pathogenic Pseudomonas aeruginosa and Staphylococcus aureus.

Abstract

ContextThe plant Moringa oleifera Lam (Moringaceae), generally termed as drumstick tree, and Citrus sinensis Linn (Rutaceae) fruit have the ability to treat multiple human infections. A biofilm is none other than a complicated microbial community whose nature is greatly resistant to antimicrobial elements. The development of biofilms in abiotic and biotic surfaces has a connection with higher levels of mortality and morbidity. Along with that, it is regarded as a vital element of bacterial pathogenicity.AimThe present study evaluated the inhibitory effect and anti-biofilm activity of Moringa oleifera (M. oleifera) and Citrus sinensis (C. sinensis) extracts against those of pathogenic Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus).Materials and methodsTwo plant materials were collected from the local market of Tabuk city and two human pathogenic microbial strains were used in the study: S. aureus and P. aeruginosa. Further, a series of morphological, physiological, and conventional biochemical tests were performed to identify the selected microorganisms. In addition to this, the study conducted the following tests: antibiotic sensitivity test, extended-spectrum β-lactamase (ESβL), and methicillin-resistant Staphylococcus aureus (MRSA) production, biofilm formation in 96-well microtiter plates, minimum inhibitory concentration (MIC) determination, the effect of sub-MICs of C. sinensis extract and M. oleifera extract on the viability of test bacteria, and finally, measurement of the inhibition of biofilm.ResultsA remarkable result of the research is that the peel extract of C. sinensisand the flesh extract of M. oleifera efficiently inhibited biofilm formation by the addition of sub-inhibitory concentrations of (1/16 x MIC - 1/2 x MIC) MRSA and ESBL, respectively. P. aeruginosa shows high resistance to piperacillin (85.0%). Similarly, the resistance of MRSA was also high (65%) against gentamycin and amikacin antibiotics. Regarding ESBL, 12 (60%) isolates showed confirmed positive and 45% of S. aureus showed MRSA activity. On observing the 12 ESβL-positive P. aeruginosa, it was found that five strains (PS1, PS4, PS6, PS8, and PS11) have formed strong biofilm, methicillin-resistant S. aureus while four strains showed strong biofilm activity (SA2, SA4, SA5, and SA8). The MIC of C. sinensis extract and M. oleifera extract against strong biofilm producers had a range of 50-2000 µg/ml concentration after overnight incubation. The study results revealed that the antibiofilm activity comparatively showed the extract of M. oleifera was better than C. sinensisagainst the mixed culture (PS1+SA8, PS6+SA2, and PS8+SA4). Hence, it is recommended to use M. oleifera as an option to monitor the development of microbial biofilms or as a model for looking for better medicines.ConclusionThe presence of antimicrobial activity found in M. oleifera and C. sinensis extracts offers convincing evidence of their likely action as antimicrobial metabolites against the studied microorganism. Anti-biofilm assay findings have shown that M. oleifera and C. sinensis extracts have effectively blocked MRSA and ESBL development in the biofilm matrix.