Screening of Antibacterial Activity In vitro of Styrax officinalis L. Covers of Berries Extracts

 

Oussama Mansour¹, ManalDarwish², Ghenwaismail³, Eman Ali², Alhasan Ali²

¹Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Al AndalusUniversity,Tartous, Syria.

²Department of Pharmacognosy, Faculty of Pharmacy, Al Andalus University, Tartous , Syria.

³Department of Pharmaceutics, Faculty of Pharmacy, Al Andalus University, Tartous  , Syria.

 

ABSTRACT:

This study is conducted to evaluate the antimicrobial activity of one extract of Styrax officinalis L envelopes of berries plant belonging to the Styracae family collected from the coastal area surrounding the city of Lattakia, Syria. Many efforts have been made to discover a new antimicrobial from a variety of sources such as micro-organisms, animals and plants. One such source is Folk medicine. Systematic screening of them may result in the discovery of a new effective compound. These extracts are prepared with polar solvent (methanol) by the steeping extraction method. The antimicrobial activity of the extracts is then assessed using the disk diffusion method against gram-positive (Staphylococcus aureus), and gram-negative (Escherichia coli) bacteria. The methanolic extracts of Styrax officinalis L envelopes of berries have shown antimicrobial activities against the tested bacteria. The activity is compared with potent broad spectrum antibiotics such as Cefipime, Amikacine and Ceftriaxon.

 

 

INTRODUCTION:

Anti-microbial agents are undeniably one of the most important therapeutic discoveries of the 20th century. However, mankind is now facing a global problem of emerging resistance in virtually pathogens¹. Concern has been expressed about the rising prevalence of pathogenic microorganisms, which are resistant to the new antibiotics that have been produced in the last three decades².

 

The toxic side effects and high cost of the new generation antibiotics with limited effective span have resulted in an increase in morbidity and mortality. Therefore, there is a crucial need to look for substances from other sources with proven antimicrobial activity³. For over thousands of years, natural plants have been considered a valuable source of medicinal agents with proven potential of treating infectious diseases and with lesser side effects compared with the synthetic drug agents. The World Health Organization estimates that 80% of people in developing countries (65% of the world’s population) still rely on traditional medicine^4,20. Plants extracts have been rich sources of medicines because they produce a host of bioactive molecules, which probably evolved as chemical defenses against infection. Most active compounds in these extracts remain unidentified, and their presence is only detected by biological tests⁵. The Styracaceae contains 11 genera and approximately 160 species consisting of small trees and shrubs, mostly native to tropical and subtropical regions. This family is well-known by the genus Styrax, which is notorious due to the production of resinous material, a pathological product, harvested by making incisions into the trees bark⁶. Styrax officinalis L. (Styracaceae) is a shrub found in Central America, Mexico and the Mediterranean region including Turkey. Its resin was used by Romans, Egyptians, Phoenicians and Ionions as incense and in therapeutics⁷.

 

A lot of research has been conducted with the aim of studying the biological activities of medicinal plants and using them for the treatment of microbial infections as possible alternatives to chemically synthetic drugs to which many infectious microorganisms have become resistant⁸. This study aims at evaluating the antimicrobial property in vitro of methanolic  extract of Styrax officinalis L cover of berries.

 

MATERIAL AND METHODS:

Styrax officinalis L. was collected from the coastal surrounding, Lattakia, Syria during the fruiting stage in 2014. The study was carried out at the Departments of Pharmacognosy and Biology, Faculty of Pharmacy, Al Andalus University, Tartous. Syria.

 

Plant Extracts:

The used part of the plant material (covers of the berries) was dried in an oven at 40˚for one hour every day for a week until the  stability of weight is achieved and then grounded to a fine powder in a mechanic grinder. The powdered plant materials (30g) were then extracted with (300ml) of  methanol 70% for 72 hours by the steeping method 3,9 , the extracted fraction was (1g:10 ml). Following filtration with Whatman filter paper ( No 1), all extracts were concentrated and evaporated to dryness at room temperature. The yields from the different extracts were weighed and dissolved in dimethyl sulphoxide (DMSO) to form the basic solution (500mgl/1ml) for each extract, then two dilutions (50,100 mg/ml) were made from each basic solution. All extracts were maintained at +4˚C until being used for disk diffusion assay⁹.

 

Test Microorganisms:

The extracts inhibitory effects were tested against two microbial species including Escherichia coli, Staphylococcus aureus obtained from Al Assad Hospital, Lattakia. The bacterial culture of test organisms were maintained on Muller Hinton Agar at 4˚C, and were subcultured in petriplates (90mm) prior to use¹⁰. DMSO was used as a negative control under the same condition for the tested microorganisms. Ceftriaxone ,Cifipime and Amikcine30μg/disc were used as positive control for the tested microorganism. The tests were carried out in duplicate. Antimicrobial activity was evaluated by measuring the zone of inhibition (mm) against the tested microorganisms¹¹.

 

Determination of Antimicrobial activity:

The antimicrobial activities of the methanol extracts by the different concentration as well as the positive control Amikacine, Cefipime and Ceftriaxon were tested by means of the disk diffusion assay against two human pathogenic bacterial strains, including Gram positive (S. aureus) and Gram negative (E. coli)¹². Approximately two cultures from each bacterial stain were inoculated over the surface petri plates containing Muller Hinton Agar using sterile swab sticks. Whatman paper disk injected by 20 µl of each concentration and ceftriaxon and amikacin disks were placed on the surface of petri plates. The plates were incubated at 37˚C for 24 hours¹³. The inhibition zones were calculated by measuring the diameters (mm) of inhibition in (including disk). Experiments were performed in duplicate and inhibition zones were compared with the positive control¹⁴.

 

RESULTS:

The anti-bacterial activities of fruit extracts were evaluated in vitro against E. coli, S. aureus, which are known to cause common infectious diseases. The results of antibacterial activity were recorded as zone of inhibition in mm around the active extract against the test microorganisms compared with the standard antibiotic Ceftriaxone, Cefipime and Amikacin as positive controls¹⁵. Methanolic extract of Styrax  officinalis have shown antibacterial activity against test organisms. The diameters of zone inhibition of this extract against S. aureus were (11-14mm) compared with the diameters of zone inhibition of Cefipime (16mm) and Ceftriaxone (12 mm). The diameters of zone inhibition of the extract against E. coli were (12-13mm) compared with the diameters of zone inhibition of Amikacine (16mm) and cefepime (8mm).

 

As shown in table 1, all concentrations of methanolic extract have shown good inhibitory effects against the tested bacteria.

 

Table 1: Antibacterial activity of metanolic extract of  the fruits of Styrax officinalis

Metanolic extracts mg/ml

(-) refer to not used

 

DISCUSSION:

Styrax officinalis is known to contain saponins, polyphenols, egonolsand triterpine. Saponins are a major family of secondary metabolites that occurs in a wide range of plant species. Saponins are steroid and triterpen glycosides so named because of their soap like properties¹⁶. Saponins have potent against microorganisms as fungi and bacteria^17,21. The antibacterial properties of saponins are generally ascribed to the ability of these molecules to complex with sterols in bacteria membrane, thus casing pore formation and loss of membrane integrity¹⁸.In addition S. officinalis contain several egonols compounds which have been mentioned in many articles and research to show the antibacterial properties¹⁹ .The presence of both saponins and lignans compounds in the covers of berries extract of Styrax officinalis is the main reason for the antibacterial activity demonstrated by some covers of extract because these constituents are dissolved in polar solvents as methanol. These extracts have shown the best antibacterial activity against the tested bacteria.

 

CONCLUSION:

The extracts of Styrax officinalis fruits are investigated individually for antimicrobial activity by disc diffusion method against selected species of Escherichia coli and Staphylococcus aureus to find the inhibitory activities of microbes. The methanolic extracts show considerably good activity. This result was compared with standard antibiotic (Ceftriaxon, Cefipim and Amikacine). In conclusion, Styrax officinalis can be supplemented in the treatment of infectious diseases and fruits extracts of the plant may have the potential to be used as antibacterial agents for searching new medicines.

 

ACKNOWLEDGEMENT: 

I would like to thank Mr. Ayham Aljghami, Instructor at The Higher Institute of Languages-Tishreen University-Syria, for the language assistance provided during the writing process of this article.

 

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Received on 27.06.2015          Modified on 10.07.2015

Accepted on 23.07.2015        © RJPT All right reserved