Croton macrostachyus. Euphorbiaceae. Hochst. ex Ferret et Galinier. C. macrostachys: A representative tree at. Kakuzi Ranch, Kenya. (Paul K.A. Konuche). Croton macrostachyus Hochst. ex Delile is a species of the genus Croton L., Euphorbiaceae family, commonly known as the spurge family. In Kenya, leaves and roots from Croton macrostachyus are used as a traditional medicine for infectious diseases such as typhoid and measles.
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In Kenya, leaves and roots from Croton macrostachyus are used as a traditional medicine for infectious diseases such as typhoid and measles, but reports on possible antimicrobial activity of stem bark do not exist.
In this study, the antibacterial and antifungal effects of methanol, ethyl acetate and butanol extracts, and purified lupeol of C. The most promising broad scale antimicrobial activity against all the studied pathogens was shown by the ethyl acetate extract.
The ethyl acetate extract induced the macfostachyus of inhibition between The antibiotic controls amoxicillin, ciprofloxacin, ampicillin, benzylpenicillin, crotpn, and cefotaxime showed antimicrobial activity with zones of inhibition within The present results give scientific evidence and support the traditional use of C.
We show that C. In many developing countries antibiotic resistance, adverse drug reactions, and the high costs of antimicrobials have made management of infectious diseases ineffective macrostcahyus 23 ]. Natural products of higher plants may be a source of new antimicrobial agents with possibly novel mechanisms of action [ crogon4 ].
The plant Croton macrostachyus has been traditionally used to treat diseases in Kenya [ 5 ].
Stem barks and twigs of Croton spp. There are both negative and positive reports on the antibacterial activity of methanol extracts from C.
Contrasting results could be attributed to the locality of plant species, parts used, time of collection, storage conditions, and methods of analysis [ 11 ]. In the present study we tested the antimicrobial activity of methanol, ethyl acetate, and isobutanol extracts from C.
The ethyl acetate extract was submitted for further purification using column chromatography fractionation and the purified compound was identified by NMR spectroscopy. Agar well diffusion method [ 12 ] was used to determine the zones of inhibition and the MIC values. The fresh stem bark was cut into small pieces using a pen knife. The cut bark was air-dried in a shaded area for three weeks. The air-dried bark was powdered using a mechanized hand grinder.
The soaked extract was separated from the plant residue using a Buchner funnel. Thin layer chromatography was used to detect the presence of polar compounds. In order to isolate pure compounds from the ethyl acetate extract silica gel column chromatography was employed. The column was washed with hexane and toluene and the sample eluted and collected with ethyl acetate. The eluted samples were dried in air and crystallized. The crystals were analyzed by NMR spectroscopy.
All experiments were referenced to the solvent peak. The following bacterial strains were used for the study: The microbial freeze-dried strains were stored in the deposit of the University of Baraton and regenerated in brain heart infusion broth HiMedia Laboratories Pvt. Antimicrobial activity of the extracts was analyzed using agar well diffusion assay according to the technique described by Taye et al.
The selection of antibiotics and antifungal agents was based on the general knowledge of the typical susceptibilities of the chosen indicator microorganisms, and the expected sensitivities were confirmed during the actual experiments. The zones of inhibition mm were recorded from measurements of the clear zones around the agar wells. The minimum inhibitory concentrations were determined using the tube dilution method. Minimal inhibitory concentration was the lowest concentration showing clear zone of inhibition.
For each solvent extract and each organism, three different experiments were conducted, giving nine readings per extract for each organism. Positive antibiotic controls were included in the assays at their recommended effective concentrations.
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Results of the growth inhibition experiments are shown in Table 1. The isobutanol extract showed activity against S. The methanol extract was active against E. The methanol extract was not active against L. All the extracts had quite equal activity against E. The antibiotic controls were constantly high. The DMSO negative control had no inhibitory activity.
Zone of inhibition of the different C. The experiments were carried out using triplicate samples. For the active extracts MIC values were determined Table 2. The methanol extract against E. Lupeol had the lowest MIC for K. NMR analysis of the fractionated and crystallized ethyl macrlstachyus extract was carried out. The chemical shifts of the carbon atoms were compared to the literature information Table 3 [ 13 ].
The chemical shifts showed that the purified compound was triterpene lupeol Figure crotoon. NMR analysis of the ethyl acetate extract from C. Chemical shifts of the numbered carbon atoms are shown.
Croton macrostachyus Hochst. ex Delile
Numbering of the carbon atoms used is that of triterpene framework. When testing methanol extracts of C. In the present study the extracts from C. The results achieved in the present study are in macrostacyhus with previous results carried out by Suffredini et al. In the present study the antibiotic controls were constantly high and some extracts induced inhibition which almost reached the antibiotic macrostavhyus, when measured as zone of inhibition.
Macrostachus results show that between the bacterial strains there is variation in susceptibility to extracts. The antimicrobial effect of the extract depends on the bacterial strain and the extraction solvent.
The broadest effect against all the studied bacteria in a dose-dependent manner was achieved by the ethyl acetate extract. The main components of C.
Because of their solubility properties one could conclude that the isobutanol and methanol extracts contained mixtures of these compounds. The active components of the isobutanol and methanol extracts remain here unknown.
Lupeol isolated from other plants has been reported to inhibit the growth of several types of bacteria, fungi, and viral species [ 14 — 20 ]. In the present study lupeol was isolated from C.
The previous reports support our finding on antimicrobial activity of lupeol. In our study, it was the first time when lupeol was extracted from the stem bark of C. The present study gives scientific evidence for the use of the extracts and especially lupeol from C. They could be utilized as preventive agents in disinfectants or submitted to further process for drug or dietary supplement development.
The provision of macrostacnyus and effective traditional medicines could offer increasing access to health care [ 1 ]. Various studies have established that herbal kacrostachyus can be developed as safe, effective, and less costly alternatives to the current medicines to treat certain bacterial infections [ 21 ]. National Center for Biotechnology InformationU. Journal List J Pathog v. Published online May Author information Article notes Copyright and License information Disclaimer.
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Abstract In Kenya, leaves and roots from Croton macrostachyus are used as a traditional medicine for infectious diseases such as typhoid and measles, but reports on possible antimicrobial activity of stem bark do not exist.
Materials and Methods 2. Plant Collection and Extraction C. Preparation of the Bark Extract The fresh stem bark was cut into small pieces using a pen knife. Bacterial Strains and Culture Conditions The following bacterial strains were used for the study: Agar Well Diffusion Method Antimicrobial activity of the extracts was analyzed using agar well diffusion assay according to the technique described by Taye et al.
Growth Inhibition Results of the growth inhibition experiments are shown in Table 1. Table 1 Zone of inhibition of the different C. Open in a separate window.
In negative control DMSO no inhibition was found. Table 3 NMR analysis of the ethyl acetate extract from C. Carbon number Sample ppm Lupeol ppm 28 Crotn When testing methanol extracts of C.
Acknowledgments This study was funded by the Academy of Finland Grant no. Nuclear magnetic resonance CDCl 3: Competing Interests The authors declare that they have no competing interests. World Health Organization; WHO traditional medicine strategy — The challenge of antibiotic resistance; need to contemplate. Indian Journal of Medical Research. Screening of Tanzanian medicinal plants for anti- Candida activity.
Evaluation of antibacterial properties of Iranian medicinal-plants against Micrococcus luteus, Serratia marcescens, Klebsiella pneumoniae and Bordetella bronchoseptica.