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Adams, R. Allured, Carol Stream, Illinois, and USA. ISBN: 0——42—1 Download references. Authors would like to thank the researcher of the herbarium of Medicinal and Aromatic Plants and Traditional Medicine Research Institute for taxonomically authenticating the plants. Department of Horticulture, Faculty of Agriculture, University of Khartoum, Shambat, , Khartoum, Sudan.

Horticultural Crops Research Center, Agricultural Research Corporation, Shambat 30, Khartoum, Sudan. Medicinal and Aromatic Plants and Traditional Medicine Research Institute, National Centre for Research, , Khartoum, Sudan.

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Reprints and permissions. Mohamed, E. Antimicrobial activity, phytochemical screening of crude extracts, and essential oils constituents of two Pulicaria spp. growing in Sudan. Sci Rep 10 , Download citation. Received : 16 June Accepted : 29 September Published : 13 October Anyone you share the following link with will be able to read this content:.

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Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily. Skip to main content Thank you for visiting nature. nature scientific reports articles article. Download PDF. Subjects Microbiology Plant sciences. Abstract The search for plant extracts with highly antimicrobial activity has been increased nowadays.

Introduction The search for substances with highly antimicrobial activity has been one of the most intensive field of research to minimize the risk of infectious diseases that caused by bacteria, fungi, viruses, and parasites, which are pathogenic to humans.

Results and discussion The yield percentages of the crude methanolic extracts and the essential oils Data in Table 1 show the yield percentages of the crude methanolic extracts, and the essential oils.

Table 1 The yield percentages of the crude methanolic extracts and essential oils of P. Full size table. Table 2 Antibacterial activity of P. undulata methanolic crude extracts, and the essential oils against the bacterial microorganisms. Table 3 Antifungal activity of P. undulata methanolic crude extracts, and the essential oils against the fungal microorganisms.

Table 5 Preliminary phytochemical screening of the methanolic crude extracts of P. Table 6 The chemical composition of the essential oil of P. Table 7 Chemical composition of the essential oil of P. Conclusions This study showed that the essential oils and the methanolic crude extracts of P.

Methods This study was carried out at the Medicinal and Aromatic Plants and Traditional Medicine Research Institute, National Center for Research, Khartoum, Sudan. Plant material Plants were selected randomly followed by antimicrobial assays. Preparation of the plant materials P.

Preparation of the crude extracts P. Preparation of the essential oils P. Antimicrobial activity screening of the methanolic crude extracts, and the essential oils of P. undulata, microorganisms The antimicrobial activity of P.

Preparation of the microorganism culture All the test microorganisms were inoculated on blood agar, and nutrient agar plates. Determination of antimicrobial activity of the methanolic crude extracts, and the essential oils of P.

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article information Zahedan Journal of Research in Medical Sciences: Vol. published online: November 30, article type: Research Article.

received: December 18, revised: July 18, accepted: October 7, Navigate to. Antimicrobial Activity of Some Medicinal Plant Extracts against Multidrug Resistant Bacteria authors:. Abstract Background: Nowadays, it is necessary to discover new and efficient antifungal or antimicrobial drugs because of increasing drug resistance organisms.

Using medicinal plants for natural treatment of diseases caused by bacterial origin has mainly been considered. Objectives: In this study, the impacts of antimicrobial medicinal plants extract were compared based on four bacteria in vitro.

Methods: In this experimental study, disc diffusion assay and the minimum inhibitory concentration MIC method were used to investigate the antibacterial effects of selected plant extract elicited by two different solvent on S. Results: The hydro-alcoholic extract of Myrtus communis myrtle and water extract of Cinnamomun zeylanicum cinnamon were the most active extracts screened for antimicrobial activities against different four bacteria as tested organisms.

The diameter of inhibition zones ranged from 23 to 28 mm. Comparison of the antibacterial effect of plant extracts and commercial drug revealed that the size of inhibition zone of penicillin against Staphylococcus aureus bacterium was larger than the plant extracts.

Petroselinum crispum parsley , Nerium oleander Oleander and Glycyrihiza glabra licorice were found to have the least effect on the tested bacteria.

Cytoplasmic pH pH int was carried out as described by Molina-Gutierrez et al. Briefly, EC and SA bacterial cells were cultured in nutrient broth at 37°C, rpm for 24 h, followed by subculturing at the same conditions for 3 h.

Cells were harvested by centrifugation at × g , 4°C for 5 min, washed twice using potassium phosphate PP buffer 50 Mm, pH 7. Afterward, the bacterial cells were incubated at 37°C, rpm for 30 min in the presence of 1.

The cells were centrifuged at × g , for 5 min and resuspended in PP buffer pH 7 , followed by addition of 10 mM glucose solution and incubated for an additional 30 min at 37°C. The cells were centrifuged at × g for 5 min, washed once and resuspended in PP buffer. Stained cells were then aliquoted into equal volumes for the control and treatment groups and the cell-free filtrate.

Fluorescence intensities were determined after 10 min by fluorescence spectrophotometer HITACHI , Japan , using excitation wavelengths of nm and emission wavelengths of nm. While the excitation slits widths were 5 nm and the emission slits widths were 10 nm.

For the control samples and bacteria free filtrate, 1 ml of PP buffer 50 mM, pH 7. Fluorescence for bacterial cells was determined by subtracting fluorescence of the respective cell-free filtrate from the treated or control groups.

DiBAC4 3 dye was used as described by Sanchez et al. Briefly, EC and SA bacterial cells were grown in nutrient broth at 37°C, rpm for 3 h. After harvesting by centrifugation at × g for 5 min, cells were washed using PP buffer 50 mM, pH 7.

After that, 1. Stained cells were aliquoted into control, treatment groups and cell-free extracts. For control groups, 1 ml of PP buffer was added to 1 ml of bacteria or cell-free filtrate. Fluorescence intensities were determined by fluorescence spectrophotometer HITACHI , using an excitation wavelength of nm and an emission wavelength of nm.

While the excitation slits widths were 5 nm and the emission slits widths were 10 nm, at room temperature 25°C. Background fluorescence resulting from the extracts added to the medium was determined. The data represent mean of three replicates ± standard deviation SD. The extraction yield obtained by conventional method showed low percentage yield Table 1 compared to those using ultrasound method.

The water extract of roselle Similarly, for ethanolic extract, the highest yield It has been reported previously that the water extract of different plants usually yields significantly higher amounts compared to ethanolic extracts of same plants Caleja et al. This may be due to using a high temperature for 30 min during extraction and also to the higher polarity of water Dhanani et al.

In addition, the utilization of vibrations to rupture plant cell walls, resulted in releasing of compounds and molecules into the solvent Tiwari, In this method, the thermal treatment is not applied, which helps protect the functional particles and increasing the recovered materials from the sample Altemimi et al.

It is highly recommended to use ultrasound method for the extraction of compounds from various sources and for different uses Grassino et al. The results revealed that the ethanolic and water extract of selected plants are efficiently suppressing the growth of food pathogens and spoilage microorganisms with variable potency.

As stated in Table 2 and Figure 1 , ethanolic extract of roselle had the maximum zone of inhibition against PA The ethanolic extract of rosemary exhibited inhibitory effect against four of the pathogenic strains EC, SE, BC, and SA while aqueous extract of rosemary was effective against three strains only EC, BC, and SA.

In the antifungal analysis, just ethanolic extract of clove and thyme had valuable results against CA with inhibition zone The antibacterial activities of ethanol extract from five plants against Listeria monocytogenes , SA, and SE in raw pork by counting bacterial enumeration were investigated, and the results confirmed that fewest colonies of tested bacteria were observed with clove extract Liu et al.

Burt demonstrated that the antimicrobial properties of thyme are owing to its content of thymol that could bind to membrane proteins by hydrophobic bonding and hydrogen bonding, and thus changing the permeability of the membranes. FIGURE 1.

Significant antimicrobial effects, expressed as MIC of each plant extract against test microorganism is given in Table 3.

The data revealed variability in the MIC among plant extracts, the lowest MIC values 0. As put forward by Mostafa et al. In general, the ethanolic extracts had lower MIC values than most of the corresponding aqueous extracts.

This lends support to previous findings in the literature that alcoholic extracts display higher antimicrobial activity than aqueous extracts Al-Hashimi, Moreover, it has been reported that large number of different chemical compounds such as phenolic compounds and its derivative compounds, the esters of weak acid, fatty acid, terpenes, and others are presented in ethanolic extracts of spice, and thus these chemical components can affect multiple target sites against the bacterial cells Burt, ; Oonmetta-aree et al.

Similar observations for MIC values, with minor variations, were observed in other studies Thuille et al. TABLE 3. The Minimum Inhibitory Concentration MIC of plant extracts against the test microorganisms. It is fundamental to develop a better understanding of the antimicrobial mechanism of plant crude extracts on the spoilage and pathogenic microorganisms.

Therefore, the effect of plant extracts on cytoplasmic pH int and membrane potential of Gram-positive SA and Gram-negative EC strains were determined.

Ethanolic extracts of clove and water extracts of thyme had the most substantial effect on both SA and EC strains; however, the water extracts of rosemary had the lowest impact compared to other extracts.

The changes that occurred in the pH int indicate damage to the bacterial cell membrane Sanchez et al. These results are consistent with Lambert et al.

Taken as a whole, the fluorescence intensity of the dye inside cells is dependent on the cytoplasmic pH of the cells where low pH results into low fluorescence intensity Aono et al. FIGURE 2. Effects of aqueous and ethanolic extracts of Roselle, Clove, Rosemary, and Thyme on the cytoplasmic pH pH in x t of Escherichia coli EC and Staphylococcus aureus SA.

Bars represent the standard deviation. On the other hand, the changes in the membrane potential of SA and EC were determined after treatment with plant extracts using DiBAC 4 3 a fluorescent dye. Suzuki et al. Our results showed a decrease in the fluorescence intensity of stained cells, which indicated displayed cell membrane hyperpolarization Figure 3.

Hyperpolarization has been suggested to be one of the primary indicators of membrane damage of bacteria cells Vanhauteghem et al. These findings match with Sanchez et al. Being an anionic oxonol dye, DiBAC 4 3 has higher permeability and accumulation in cells with polarized membrane potential due to the presence of higher positive charges inside the cytoplasmic membrane Joux and Lebaron, On the other side, hyperpolarization results in poor uptake and thus reduce the accumulation of the dye inside the cytoplasmic membrane of the bacteria leading to low fluorescence intensity Sanchez et al.

FIGURE 3. Effects of aqueous and ethanolic extracts of Roselle, Clove, Rosemary, and Thyme on the membrane potentials of Escherichia coli EC and Staphylococcus aureus SA. Based on the results, it can be concluded that using ultrasound method during extraction effectively improve the extraction yield.

Overall, water and ethanolic extracts from selected plants possess antimicrobial activity as they could inhibit the growth of tested food pathogens and spoilage microorganisms.

The ethanolic and aqueous extracts of roselle and clove had antimicrobial activity against all tested microorganisms except for the CA that was affected only by ethanolic extracts of clove and thyme.

A decrease in cytoplasmic pH pH int and cell wall disruption was observed in cells treated with plant extracts, suggesting a possible mechanism of antibacterial action.

These findings indicate that the plant extracts tested in this study could be used as natural preservative agents in food to eliminate or control the growth of spoilage and pathogenic microorganisms.

FG is a student and was responsible for conducting the antimicrobial assays and contributed to the manuscript revisions mainly materials and methods. JL contributed to the development of the experimental design, data analysis, the manuscript draft, and proofreading. WM contributed to the experimental design, plants extraction, interpreted the results, as well as data analysis, and interpretation.

JX and FC carried out the antimicrobial mechanism experiments, helped to write the manuscript. MC and SH were the lead investigators, designed the study, conducted the experiments, supervised the students, drafted the manuscript, and final proofreading.

This work was funded by the projects granted by National Natural Science Foundation of China NSFC and NSFC , Zhejiang Province Science Funding LQ15C and the Science Plan Project of Food and Drug Supervision System in Zhejiang Province BH The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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