Effect of Fractioning on Antibacterial Activity of Enantia chlorantha Oliver (Annonaceae) Methanol Extract and Mode of Action
Effect of Fractioning on Antibacterial Activity of Enantia chlorantha Oliver (Annonaceae) Methanol Extract and Mode of Action
Rebeca Ebelle Etame,1 Raymond Simplice Mouokeu,2 Cedric Laurel Cidjeu Pouaha,3 Igor Voukeng Kenfack,4 Raphael Tchientcheu,3 Jean Paul Assam Assam,3 Frank Stève Monthe Poundeu,3 Alembert Tchinda Tiabou,1 François Xavier Etoa,5 Jules Roger Kuiate,4 and Rosalie Anne Ngono Ngane3
1Institute of Medical Research and Medicinal Plant Studies (IMPM), P.O. Box 6163, Yaoundé, Cameroon
2Institute of Fisheries and Aquatic Sciences, University of Douala, P.O. Box 7236, Douala, Cameroon
3Faculty of Sciences, University of Douala, P.O. Box 24157, Douala, Cameroon
4Faculty of Sciences, University of Dschang, P.O. Box 67, Dschang, Cameroon
5Faculty of Sciences, University of Yaoundé I, P.O. Box 812, Yaoundé, Cameroon
Correspondence should be addressed to Raymond Simplice Mouokeu; rf.oohay@isyarom
Received 28 November 2017; Revised 19 March 2018; Accepted 1 April 2018; Published 29 April 2018
Academic Editor: Nativ Dudai
Copyright © 2018 Rebeca Ebelle Etame et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Infectious diseases caused by bacteria constitute the main cause of morbidity and mortality throughout the world and mainly in developing countries. In this work, the influence of fractioning and the mode of action of stem barks methanol extract of Enantia chlorantha were investigated. The aim was to optimize the antibacterial activity of the methanol extract. The extract was prepared by maceration of barks powder in methanol. Fractioning was done using increasing solvents polarity. Standard phytochemical methods were used for phytochemical screening. Minimum Inhibitory Concentrations (MIC) and Minimum Bactericidal Concentration (MBC) of the methanol extract and fractions were determined using broth microdilution method. The studied mode of action of both methanol extract and n-butanol fraction included antibiofilm activity, H+-ATPase-mediated proton pumping assay, salt tolerance, and cells cycle. The methanol extract of E. chlorantha stem barks was found to be active on all the bacteria tested (32 ≤ MIC ≤ 512 μg/mL), its activity being significant (MIC < 100 μg/ml) out of 5 of the 28 clinical isolates used. Salmonella enterica serovar paratyphi A was the most sensitive (32 μg/mL). Compared to the extract and other fractions, the n-butanol fraction was found to be more active (32 ≤ MIC ≤ 256). Significant antibacterial activity of this fraction was observed out of 10 of the 28 bacterial isolates and 3 out of 7 bacterial strains. Lowest MIC values (32 μg/ml) of this fraction were obtained with Escherichia coli (136), Pseudomonas aeruginosa (CIP 76110), and Salmonella enterica serovar typhi 9. The methanol extract of E. chlorantha and its n-butanol fraction revealed several modes of action including the prolongation of the latency phase of the bacterial growth, the inhibition of the pump with protons H+ - ATPases bacterial, the loss of the salt tolerance of the Staphylococcus aureus, and inhibition of the formation of the bacterial biofilm. The present results showed that the n-butanol fraction of the methanol stem barks extract of E. chlorantha possess the essential antibacterial components and could best be used to fight against bacterial infections as compared to methanol extract.
1. Introduction
Infectious diseases caused by bacteria constitute a public health problem all over the world and particularly in Africa where living conditions are precarious [1, 2]. The discovery of antibiotics has been a real relief for humanity because they have greatly reduced the incidence of infectious diseases [3]. The often inappropriate prescription of antibiotics and their inappropriate use by the population among others have led to selection pressure by bacteria [4]. This has led, for several decades, to the emergence of resistant bacteria, which is the main cause of increased frequency of therapeutic failures, mortality, and high costs of treatment [5]. Alternative remediation could include new antimicrobial substances that are effective, available, and low in toxicity. Medicinal plants are a credible research pathway. Indeed, they are rich in molecules of an exceptional chemical variability making them a vast reservoir of substances that can act through various modes of action [6]. Many approaches have so far been used to demonstrate the antibacterial activity of plants. They include crude extract preparation using variety of solvents [7], purification of active compounds from extracts [8–11], successive extraction with solvents with increasing polarity [12], and distillation. Solvent extraction has been the most used method but recent studies established increased activity with fractionation of crude extract [13, 14]. In ad (...truncated)