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      Antimicrobial, antioxidant, toxicity and phytochemical assessment of extracts from A cmella uliginosa, a leafy-vegetable consumed in Bénin, West Africa

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          Abstract

          Background

          Acmella uliginosa (Asteraceae) is a flowering plant whose leaves are consumed as a vegetable in Benin. They are also traditionally used as an antibiotic in the treatment of infectious diseases. To evaluate the therapeutic potential and toxicity effect of this leafy-vegetable, the antibacterial, antifungal, antioxidant activities and, toxicity and phytochemical constituents were investigated.

          Methods

          Dichloromethane, methanol and aqueous extracts of Acmella uliginosa were evaluated for their antimicrobial activity against six bacterial and six fungi strains. Antibacterial and antifungal activities were investigated by microdilution method and agar diffusion method respectively. Antioxidant activity was assessed using the 2,2-diphenyl-1-picryl-hydrazyl assay and phytochemical screening was carried out using standard procedures. Finally, oral acute toxicity at a dose of 2000 mg/kg was done according to the Organization for Economic Co-operation and Development guideline n° 423.

          Results

          The antibacterial activity was broad spectrum, inhibiting both Gram-positive and Gram-negative bacteria. The minimum inhibitory concentration ranged from 0.625 to 5 mg/ml. The antifungal evaluation show that all the extracts inhibited mycelial growth and sporulation of fungi with percentages of inhibition ranging from 9.39 to 75.67 % and 22.04 to 99.77 %, respectively. In DPPH radical scavenging assay, the effect on reducing free radicals increased in a dose dependent manner. The percentage of inhibition of DPPH ranged from 0.94 to 73.07 %. Phytochemical screening revealed the presence of coumarin, flavonoid, naphtoquinone, anthracene derivative, saponin, lignan, triterpene and tannin. The dichloromethane and methanol extracts showed the best biological activities; they were also shown as the best extraction solvents of phytochemicals. In the acute toxicity evaluation, all animals were physically active and no deaths of rats were observed during the test. However, the aqueous extract promoted biochemical, hematological and histopathological alterations of treated rats at 2000 mg/kg body weight.

          Conclusion

          A. uliginosa extracts contains antimicrobial, antioxidant agents and was not lethal for rats when ingested. However, according to the results obtained for biochemical, hematological, and histopathological analysis, caution is required regarding its consumption.

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          Most cited references43

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          Antimicrobial activity of flavonoids

          Flavonoids are ubiquitous in photosynthesising cells and are commonly found in fruit, vegetables, nuts, seeds, stems, flowers, tea, wine, propolis and honey. For centuries, preparations containing these compounds as the principal physiologically active constituents have been used to treat human diseases. Increasingly, this class of natural products is becoming the subject of anti-infective research, and many groups have isolated and identified the structures of flavonoids possessing antifungal, antiviral and antibacterial activity. Moreover, several groups have demonstrated synergy between active flavonoids as well as between flavonoids and existing chemotherapeutics. Reports of activity in the field of antibacterial flavonoid research are widely conflicting, probably owing to inter- and intra-assay variation in susceptibility testing. However, several high-quality investigations have examined the relationship between flavonoid structure and antibacterial activity and these are in close agreement. In addition, numerous research groups have sought to elucidate the antibacterial mechanisms of action of selected flavonoids. The activity of quercetin, for example, has been at least partially attributed to inhibition of DNA gyrase. It has also been proposed that sophoraflavone G and (−)-epigallocatechin gallate inhibit cytoplasmic membrane function, and that licochalcones A and C inhibit energy metabolism. Other flavonoids whose mechanisms of action have been investigated include robinetin, myricetin, apigenin, rutin, galangin, 2,4,2′-trihydroxy-5′-methylchalcone and lonchocarpol A. These compounds represent novel leads, and future studies may allow the development of a pharmacologically acceptable antimicrobial agent or class of agents.
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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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              Potential of Cameroonian plants and derived products against microbial infections: a review.

              In Cameroon, infectious diseases are amongst the most commonly notified diseases and largest cause of mortality. Many plants are used locally in traditional medicine for their treatment. The aim of the present review is to summarize currently available evidence and knowledge concerning Cameroonian plants used to treat bacterial and fungal infections, and the efficacy of plant-derived extracts and compounds. The traditional uses of plants in the treatment of infectious diseases have been collected and tabulated. The antimicrobial activity of the extracts and the chemical constituents of most of these plants are summarized in this report. Plants used traditionally in Cameroonian medicine, with laboratory work on any part or products, have been documented. Numerous extracts and compounds have been tested for antimycobacterial, antibacterial and antifungal efficacy and some of them were significantly active. Most of the bioactive compounds isolated were phenolics and alkaloids. In conclusion, many plant species are used in traditional medicine in Cameroon to treat infectious diseases, and several interesting openings have originated for further inquiry following IN VITRO antimicrobial activity evaluation. However, much work is still to be done to standardize methods and cut-off points for describing the antimicrobial activity, and on the study of the mechanisms of action. © Georg Thieme Verlag KG Stuttgart · New York.
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                Author and article information

                Contributors
                llagnika@gmail.com
                chifmadjid@gmail.com
                rafatouadjileye@gmail.com
                laleyec@yahoo.fr
                ambaliou.sanni@gmail.com
                Journal
                BMC Complement Altern Med
                BMC Complement Altern Med
                BMC Complementary and Alternative Medicine
                BioMed Central (London )
                1472-6882
                27 January 2016
                27 January 2016
                2015
                : 16
                : 34
                Affiliations
                [ ]Unité de Biochimie et Biologie Moléculaire, Equipe de Biochimie et Substances Naturelles Bioactives, Faculté des Sciences et Techniques, Université d’Abomey-Calavi, Cotonou, 04 BP 0320 Bénin
                [ ]Unité de Biologie Humaine, Laboratoire de Cytogénétique et de Biologie Moléculaire, Faculté des Sciences de Santé, Université d’Abomey-Calavi, Cotonou, Bénin
                Article
                1014
                10.1186/s12906-016-1014-3
                4728828
                26817601
                b08a3f6d-4c85-4b62-88ce-7d1985beee61
                © Lagnika et al. 2016

                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
                : 22 September 2015
                : 22 January 2016
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100004413, International Foundation for Science;
                Award ID: F/5673-1
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2016

                Complementary & Alternative medicine
                acmella uliginosa,antifungal,antibacterial,antioxidant,toxicity,phytochemical

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