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      Antibacterial activities of the extracts, fractions and isolated compounds from Canarium patentinervium Miq. against bacterial clinical isolates

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          Abstract

          Background

          Canarium patentinervium leaves are used by the local indigenous people of Malaysia for wound healing. The current study is undertaken to screen the comprehensive antibacterial activity of the leaves and barks extracts, fractions and isolated compounds from this plant. Bioassay guided fractionation was also undertaken to deeply evaluate the antibacterial activity of the water fraction of the leaves extract. This is to provide preliminary scientific evidence to the ethnopharmacology usage of this plant by investigating antibacterial properties of the plant and its isolated constituents.

          Methods

          Bio-assay guided fractionation and subsequent isolation of compounds using open column chromatography. The antibacterial activity against gram positive and gram negative ATCC strain and resistant clinical strains were evaluated using microtiter broth dilution method to determine minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill assay. The chemical structure of the isolated compounds from the water fraction of the ethanol extract of leaves was elucidated using Nuclear Magnetic Resonance (NMR).

          Results

          The ethanol extract of the leaves and barks showed antimicrobial activity against all four ATCC and eight clinical isolates. The ethanol extract of the leaves and the corresponding water fraction had good activity against MRSA S. aureus. (MIC: 250 μg/ml) and had bactericidal effect on eight of the clinical strains (MSSA,MRSA, oxacillin-resistant CONS, oxacillin-sensitive CONS, Enterococcus faecalis, Klebsiela species , Kleb pneumoniae ESBL and Candida parapsilosis). Further phytochemical investigation of the water fraction of the crude ethanol extract of leaves afforded compound 7 (hyperin) and compound 8 (cynaroside) that had bactericidal activity against tested bacterial species (MIC 50 μg/ml and 100 μg/ml). The two compounds were isolated from this genus for the first time.

          Conclusions

          These results may provide a rational support for the traditional use of Canarium patentinervium Miq. in infections and wound healing, since the antimicrobial compounds isolated were also present in the leaves extract.

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          Most cited references37

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          A sensitive and quick microplate method to determine the minimal inhibitory concentration of plant extracts for bacteria.

          J Eloff (1998)
          Agar diffusion techniques are used widely to assay plant extracts for antimicrobial activity, but there are problems associated with this technique. A micro-dilution technique was developed using 96-well microplates and tetrazolium salts to indicate bacterial growth. p-Iodonitrotetrazolium violet [0.2 mg/ml] gave better results than tetrazolium red or thiazolyl blue. The method is quick, worked well with Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli and with non-aqueous extracts from many different plants. The method gave reproducible results; required only 10-25 microliters of extract to determine minimal inhibitory concentrations, distinguished between microcidal and microstatic effects, and provided a permanent record of the results. Using S. aureus, and a Combretum molle extract, the technique was 32 times more sensitive than agar diffusion techniques and was not sensitive to culture age of the test organism up to 24 hours. The S. aureus culture could be stored up to 10 days in a cold room with little effect on the assay results. This method was useful in screening plants for antimicrobial activity and for the bioassay-guided isolation of antimicrobial compounds from plants. MIC values determined for sulfisoxazole, norfloxacin, gentamicin, and nitrofuratoin were similar to values indicated in the literature but values obtained with trimethroprim and ampicillin were higher with some bacteria.
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            Distribution and Biological Activities of the Flavonoid Luteolin

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              Flavonoids are scavengers of superoxide anions.

              Seven flavonoids and three non-flavonoid antioxidants, i.e. butylated hydroxyanisole, chlorpromazine and BW 755 C, were studied as potential scavengers of oxygen free radicals. Superoxide anions were generated enzymatically in a xanthine-xanthine oxidase system and non-enzymatically in a phenazine methosulphate-NADH system, and assayed by reduction of nitro blue tetrazolium. The generation of malonaldehyde (MDA) by the ascorbate-stimulated air-oxidised boiled rat liver microsomes was considered as an index of the non-enzymatic formation of hydroxyl radicals. Flavonoids but not non-flavonoid antioxidants lowered the concentration of detectable superoxide anions in both enzymic and non-enzymic systems which generated these SOD-sensitive radicals. The most effective inhibitors of superoxide anions were quercetin, myricetin and rutin. Four out of seven investigated flavonoids seemed also to suppress the activity of xanthine oxidase as measured by a decrease in uric acid biosynthesis. All ten investigated compounds inhibited the MDA formation by rat liver microsomes. Non-flavonoid antioxidants were more potent MDA inhibitors than flavonoids. It is concluded that antioxidant properties of flavonoids are effected mainly via scavenging of superoxide anions whereas non-flavonoid antioxidants act on further links of free radical chain reactions, most likely by scavenging of hydroxyl radicals.
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                Author and article information

                Contributors
                mogana@ucsiuniversity.edu.my , rmogana76@gmail.com
                achyutraj05@gmail.com
                tzar@ppukm.ukm.edu.my
                ramliza@ppukm.ukm.edu.my
                Christophe.Wiart@nottingham.edu.my
                Journal
                BMC Complement Med Ther
                BMC Complement Med Ther
                BMC Complementary Medicine and Therapies
                BioMed Central (London )
                2662-7671
                14 February 2020
                14 February 2020
                2020
                : 20
                : 55
                Affiliations
                [1 ]GRID grid.444472.5, Department of Pharmaceutical Biology, Faculty of Pharmaceutical Sciences, , UCSI University, ; No. 1 Jln Menara Gading, UCSI Heights, 56000 Cheras, Kuala Lumpur, Malaysia
                [2 ]ISNI 0000 0001 2114 6728, GRID grid.80817.36, Central Department of Chemistry, , Tribhuvan University, ; Kritipur, Kathmandu, Nepal
                [3 ]Department of Medical Microbiology and Immunology, Hospital National University of Malaysia, Cheras, Kuala Lumpur, Malaysia
                [4 ]GRID grid.440435.2, School of Pharmacy, Faculty of Science, Center for Natural and Medicinal Products Research, , University of Nottingham (Malaysia Campus), ; Jalan Broga, 43500 Semenyih, Selangor Malaysia
                Author information
                http://orcid.org/0000-0002-5664-9293
                Article
                2837
                10.1186/s12906-020-2837-5
                7076860
                32059725
                eb8b9df4-9256-4ba4-9787-40466c14e709
                © The Author(s). 2020

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 2 August 2019
                : 30 January 2020
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2020

                canarium patentinervium miq.,bacterial clinical isolates,mic,mbc,hyperin,cynaroside

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