Endophytic actinobacteria of medicinal plant Aloe vera: Isolation, antimicrobial, antioxidant, cytotoxicity assays and taxonomic study

Objective: To explore the new sources of novel bioactive compounds having pharmaceutical and agricultural interest and to search the endophytic actinobacteria from medicinal plants. Methods: NAF-1 an endophyte actinobacteria was isolated from leaves of medicinal plant Aloe vera collected in Marrakesh, Morocco using Bennett agar as selective medium. NAF-1 was tested for its antimicrobial activity against five pathogenic bacteria such as Staphylococcus aureus PIC 53156, Micrococcus luteus ATCC381, Bacillus subtilis ATCC 14579, Pseudomonas aeruginosa DSM 50090 and Escherichia coli ATCC 8739 and four human clinic fungi belonging to the Candida, Aspergillus and Microsporum genera. Several antioxidant activities were studied such as DPPH free radical scavenging, β-carotene and linoleic acid and reducing power assays. The total of phenol and flavonoid was also calculated. Using Artemia salina shrimp assay, the cytotoxicity of NAF-1 crude extract was determined. Results: The results revealed that the actinobacteria showed a high activity(≥20 mm) against only Gram positive bacteria but it had a moderate activity(between 13 and 15 mm) against Human clinic fungi. The isolate also exhibited a LD50 of 14.20 μg/mL in the cytotoxicity assay. The result showed that the crude extract presented an interesting free radical-scavenging activity with IC50 value of(5.58 ± 0.26) μg/mL and a high value of phenolic and flavonoid compounds with(15.41 ± 0.18) μg GAE/mg extract and(11.41± 0.06) μg QE/mg extract respectively. Moreover, the taxonomic position of our endophyte actinobacteria using the morphological and physiological criteria and using 16 S r RNA gene sequence(polyphasic approach) showed that the NAF-1 isolate was similar to Streptomyces hydrogenans which was never described as an endophyte actinobacteria. Conclusions: This isolated strain appears promising resources of bioactive agents and can be exploited to produce therapeutic agents active against pathogenic disease.

In vitro antimicrobial and synergistic effect of essential oil from the red macroalgae Centroceras clavulatum (C. Agardh) Montagne with conventional antibiotics

Objective:To study the chemical profile,antimicrobial properties,and synergistic effect with known antibiotics of essential oil extracted from the marine red macroalgae Centroceras clavulatum(C.Agardh)Montagne,collected in Morocco.Methods:The chemical composition of the oil was analyzed by gas chromatography-mass spectrometry.The oil was evaluated for antibacterial(Pseudomonas aeruginosa,Escherichia coli,Bacillus subtilis,Micrococcus luteus,Staphylococcus aureus,and Klebsiella pneumoniae),and antifungal activity(Candida albicans,Candida glabrata,Candida krusei,and Candida parapsilosis),by the disc diffusion method.The minimum inhibitory and minimum microbicidal concentrations of the oil were determined,as well as the synergistic effects of its application combined with the antibiotics ciprofloxacin and fluconazole,by the checkerboard method.Results:Thirty molecules were identified in the essential oil,comprising 96.27%of the total oil composition.Monoterpenes such as carvacrol(36.06%)were the most abundant compounds,followed by caryophyllene(14.67%),endo-borneol(9.04%),pyroterebic acid(3.23%)and caryophyllene oxide(3.13%).The oil exhibited a moderate antimicrobial activity with inhibition zone diameters ranging from 9.0 to 15.0 mm.The minimum inhibitory concentration values varied between 0.9 and 14.7 mg/mL,and Bacillus subtilis and Escherichia coli were the more sensitive bacteria with 0.9 and 1.9 mg/mL,respectively.The minimum microbicidal concentration values ranged from 0.4 to 14.7 mg/mL.A significant synergic action was observed when the oil was applied in combination with ciprofloxacin and fluconazole,with fractional inhibitory concentration index values ranging from 0.31 to 0.50.Synergy was found in 80%of the combinations and a 2 to 16-fold reduction of antibiotics MIC was observed.Conclusions:Our findings suggest that the essential oil of Centroceras clavulatum should be further appraised for its potential use in the management of multi-drug resistant microorganisms,with the purpose to restore the activity of standard antimicrobial drugs.