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The evaluation of antimicrobial and cytotoxic activity of the essential oil extracted from the aerial parts of southernwood herb (Artemisia abrotanum L.) that recently grown in Iraq

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The evaluation of antimicrobial and cytotoxic activity of the essential oil extracted from the aerial parts of southernwood herb (Artemisia abrotanum L.) that recently grown in Iraq

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Objective: This research is to study the assessment of the antimicrobial and cytotoxic activity of the essential oil extracted from the aerial parts of Artemisia abrotanum L. that recently grown in Iraq.Methods: The essential oil of A. abrotanum was extracted by hydrodistillation using Clevenger apparatus. This essential oil was tested for antimicrobial activity of five different pathogenic microorganisms (Gram-positive [Staphylococcus aureus and Bacillus subtilis] and Gram-negative [Salmonella typhi and Escherichia coli] bacterial strains) and fungi: Candida albicans using diffusion well agar method. Furthermore, this essential oil was tested for cytotoxic activity using rhabdomyosarcoma cell line, and the growth or inhibition of cancer cells was measured by MTT method.Results: The obtained results show that the antibacterial activity for A. abrotanum against S. aureus was at concentrations 40, 25, and15 μl with minimum inhibitory concentrations of 20 mm, while it showed antibacterial activity against E. coli for four different concentrations of 40, 25, 15, and10 μl with inhibition zone of 16, 12, 14, and 10 mm, respectively, and it showed antifungal activity against C. albicans at four concentrations 40, 25, 15, and10 μl with inhibition zone of 18, 24, 26, and 30 mm, respectively. The cytotoxic activity of the extracted essential oil was showed that the three concentrations of the extract (25, 50, and 100 μg/ml) were all lower significantly as compared to dimethyl sulfoxide group. A significant difference was seen for group 25 with both groups 50 and 100, but no significant difference was seen between the two later. Finally, the antimicrobial and anticancer activity of this plant could be due to its essential oil constituents: Borneol, cymene, camphor, terpineol, eucalyptol, and aromadendrene.Conclusion: The essential oil of A. abrotanum L. has a potent antimicrobial and anticancer effect against the tested microbial organisms and the cancer cells.
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Vol 10, Issue 10, 2017
Online - 2455-3891
Print - 0974-2441
THE EVALUATION OF ANTIMICROBIAL AND CYTOTOXIC ACTIVITY OF THE ESSENTIAL
OIL EXTRACTED FROM THE AERIAL PARTS OF SOUTHERNWOOD HERB (ARTEMISIA
ABROTANUM L.) THAT RECENTLY GROWN IN IRAQ
SARAH S ALMAHDAWY1*, ALI MUAFFAQ SAID2, IBRAHIM S ABBAS1, ASHOUR H DAWOOD3
1Department of Pharmacognosy and Medicinal Plants, College of Pharmacy, Al-Mustansiriya University, Baghdad, Iraq. 2Department of
Pharmacy, Al-Yarmouk University College, Baghdad, Iraq. 3Department of Pharmaceutical Chemistry, College of Pharmacy, Al-Mustansiriya
University, Baghdad, Iraq. Email: sara.saad.abd@gmail.com
Received: 02 August 2017, Revised and Accepted: 28 August 2017
ABSTRACT
Objective: This research is to study the assessment of the antimicrobial and cytotoxic activity of the essential oil extracted from the aerial parts of
Artemisia abrotanum L. that recently grown in Iraq.
Methods: The essential oil of A. abrotanum was extracted by hydrodistillation using Clevenger apparatus. This essential oil was tested for antimicrobial
activity of five different pathogenic microorganisms (Gram-positive [Staphylococcus aureus and Bacillus subtilis] and Gram-negative [Salmonella typhi
and Escherichia coli] bacterial strains) and fungi: Candida albicans using diffusion well agar method. Furthermore, this essential oil was tested for
cytotoxic activity using rhabdomyosarcoma cell line, and the growth or inhibition of cancer cells was measured by MTT method.
Results: The obtained results show that the antibacterial activity for A. abrotanum against S. aureus was at concentrations 40, 25, and15 µl with
minimum inhibitory concentrations of 20 mm, while it showed antibacterial activity against E. coli for four different concentrations of 40, 25, 15,
C. albicans at four concentrations 40, 25,

 
was seen for group 25 with both groups 50 and 100, but no significant difference was seen between the two later. Finally, the antimicrobial and
anticancer activity of this plant could be due to its essential oil constituents: Borneol, cymene, camphor, terpineol, eucalyptol, and aromadendrene.
Conclusion: The essential oil of A. abrotanum L. has a potent antimicrobial and anticancer effect against the tested microbial organisms and the
cancer cells.
Keywords: Southernwood, Artemisia abrotanum        
bromide).
INTRODUCTION
The genus Artemisia L. is among the largest and most widely spread
          
        
       Artemisia abrotanum

feet, it consists of yellowish-white flowers, and has a fragrance similar
        
compounds for the development of new chemotherapeutic agents. In
vitro evaluation of plants for the antimicrobial property is the first step
toward achieving the aim for developing eco-friendly management of
infectious diseases of humans by searching for new biomolecules of

Essential oils play an important role in the biological activity of the plant;
they were mostly composed of terpenes and are obtained by means of
steam distillation, hydro distillation, or solvent extraction of different
parts of the aromatic plants [4]. A. abrotanum was traditionally used
as an antiseptic, astringent, emmenagogue, antidiabetic, expectorant,
febrifuge, stomachic, antimalarial, anti-inflammatory, vermifuge, and
spasmolytic and used for treating upper respiratory tract disease,
antibacterial, antifungal, cancer, cough, and fever [5]. In Iraq, there is no
study about A. abrotanum essential oil biological activity because this
plant was introduced recently to Iraq for decorative purposes.
METHODS
Plant collection
The plant aerial parts of A. abrotanum were collected from Iraq at
          
Mustansiriya University. The plant was authenticated by National Iraqi
       
dried in the shade at room temperature for 7 days until crisp and then
were grinded by mechanical mills and weighed (Fig. 1).
Isolation procedure
The essential oil content of A. abrotanum was extracted by hydro
distillation method by the use of Clevenger apparatus, 100 g of the plant
material were hydrodistilled by adding 500 ml of distilled water in

oil was collected after observing that there is no increase in volatile oil
 was added to the essential oil in the
cylinder to remove any water left. Then, it was kept into tightly closed,
dark, small, glass containers and kept into the refrigerator and stored at
4°C. The average percentage of volatile oil content of the plant sample

Antimicrobial activity of A. abrotanum
The essential oil of A. abrotanum was tested for antimicrobial activity of
five different pathogenic microorganisms gram positive: [Staphylococcus


Research Article

Asian J Pharm Clin Res, Vol 10, Issue 10, 2017, 384-387
Almahdawy et al.
aureus (S.aureus), Bacillus subtilis(B.subtilis)], gram negative: [Salmonella
typhi (S.typhi), Escherichia coli (E.coli)] bacterial strains and fungi:
Candida albicans (C.albicans)      
then, the growth or inhibition of bacterial cells was determined by
          


dish and allowed to solidify in laminar air flow chamber, and using sterile

plate technique, and the gel puncture method done using the sterile cork
   

of bacterial suspension was added in each plate. Beside of this, positive
control (medium with bacterium with extract) also was prepared. For
determination of minimum inhibitory concentrations (MIC), serial
dilutions of the essential oil of both species of artemisia were done using



 
by diameter.
Cytotoxic activity of A. abrotanum L. in cancer tissue culture
         
concentrations of plant extracts (essential oil) during the log phase of
growth and the effect determined after recovery time [7], all solutions
     
cell seeding. Cell suspension was prepared using 25 cm tissue culture
   
2 after detachment of the cells from the
flask surface single cell suspension by gently taping of the flask followed
        

bottom microtiter plate using automatic micropipette containing 1×105

 2   
the well for RD cell line, and the second stage (exposure stage), the cells
were then exposed to three different concentrations (25, 50, and 100 µl
        
six wells of each concentration, the cells that incubated with vehicle
or methotrexate represented the negative control and positive control,

2

drugs solutions were added, and the plate was further incubated for
2 atmosphere. Then, 50 µl of MTT
          
      2
atmosphere. The MTT is a colorimetric assay to assess cell counts and
   
         
      
   
culture medium was removed from wells, and the converted dye was
        
supplemented with 0.04N HCl). The absorbency of the wells was
measured with a microculture plate reader at 570 nm. The optical

Statistical analysis
         
         
means±standard deviation (SD). The level of significance (p<0.05) was
used for analysis of results presented in this study.
RESULTS AND DISCUSSION
The antimicrobial effects of A. abrotanum L.
The results showed in Table 1 indicated that the antibacterial activity
for A. abrotanum against S. aureus was at concentration 40, 25, and

E. coli for four different concentrations of 40, 25, 15, and 10 µl with
     
could be explained the results for A. abrotanum in Table 1 of our
work, although there are limited studies available, the plant was also
showed antimicrobial activity against both types of bacteria, in which
is compatible with the previous studies [11,12], could be due to that it
contains borneol, cymene, camphor, terpineol, and eucalyptol that have
antibacterial activity.
Furthermore, the results showed that this plant has antifungal
activity against C. albicans at four concentrations (40, 25, 15, and
   
antibacterial activity may be attributed to the essential oil constituents
such as: camphor which shows a very potent antimicrobial activity
against S.aureus and C. albicans     
activity against different types of gram positive and gram negative
organisms [14], cymene that exhibit antibacterial activity against
S. aureus, E. coli [15], Bisabolene that demonstrate the antibacterial
activity against S. aureus, E.coli, Pseudomonas aeruginosa 
aromadendrene that appear antibacterial activity against multidrug-
resistant bacterial pathogens [17], most of the essential oils that contain
        
studies are available for the antifungal effects that could explained our
results, and for A. abrotanum, the plant showed that activity against
C. albicans      
that in the essential oil of the plant which they have an antifungal action
such as borneol [20] and camphor [21], which may be explained the
results in this work.
The cytotoxic activity of A. abrotanum L.
          
below [22].
The effects of A. abrotanum on RD cell viability:
 
 
    
the three concentrations of each extract and with both negative and
positive controls.
The results showed that the group of methotrexate decreased
          
         

as compared to methotrexate group. The three concentrations of the


groups 50 and 100, but no significant difference was seen between the
two later. The results obtained in this work were compatible with the
previous studies before. The plant artemisia was showed anticancer
effects for several different species.
This anticancer activity may be due to the active constituents present in
  
Fig. 1: The effects of A. abrotanum on RD cell viability (statistical
analysis)

Asian J Pharm Clin Res, Vol 10, Issue 10, 2017, 384-387
Almahdawy et al.
        
(MCF-7) cells [24], Borneol that exhibit cytotoxic activity against
(HepG2, Caco-2 and VH10) cell lines [25], Bisabolene that demonstrate

      
      

 
for the treatment of many forms of cancer. In many situations, the actual
compound isolated from the plant may not serve as the drug but leads to
the development of potential novel agents. The ability to attach agents
to carrier molecules directed to specific tumors holds promise for the
effective targeting of highly cytotoxic natural products to the tumors

CONCLUSION
The essential oil of A. abrotanum that recently grows in Iraq extracted
by hydro distillation has a potent antimicrobial and anticancer effect
against the tested microbial organisms and the cancer cells. The
essential oil from this plant shows several biological activities, and
more researches are required to study the biologic activities for each
active constituents that obtain in this study. The essential oil shows
several biological activities, more researches are required to study the
biologic activities for each active constituents that obtain in this study,
and finally, the essential oil in this study showed marked anticancer
activity, more researches are required for further assessment of this
effect against other different types of cancers, both in vitro and in vivo.
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Table 1: Antibacterial activity of A. abrotanum
Concentrations
of essential oil of
A. abrotanum (µl)
S. aureus
(mm)
E. coli
(mm)
C. albicans
(mm)
40 20  
 - - -
25 24 12 24
20 - - -
15  14 
10 - 10 
A. abrotanum: Artemisia abrotanum, S. aureus: Staphylococcus aureus,
E. coli: Escherichia coli, C. albicans: Candida albicans
Graph 1: The effects of A. abrotanum on RD cell viability
(statistical analysis)
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... Moreover, A. abrotanum is included in homeopathic medicine according to the French Pharmacopoeia. These preparations are recommended for the treatment of the inflammation of the colon, rosacea, frostbite, inflammation of the lymph nodes, mucous membranes, and anxiety [166][167][168]. ...
... Antitumor activity was confirmed in A. abrotanum leaf extracts and essential oil components [20,168]. Flavonoids from A. abrotanum are reported to relieve the symptoms of allergic rhinitis [117]. ...
... Essential oils or extracts of Artemisia ssp. discussed in this review have antibacterial, antifungal, and antioxidant activities [14,20,38,39,58,84,85,87,88,[91][92][93]122,168,201,212,217,[232][233][234][235][236][237][238]. ...
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Artemisia species play a vital role in traditional and contemporary medicine. Among them, Artemisia abrotanum, Artemisia absinthium, Artemisia annua, Artemisia dracunculus, and Artemisia vulgaris are the most popular. The chemical composition and bioactivity of these species have been extensively studied. Studies on these species have confirmed their traditional applications and documented new pharmacological directions and their valuable and potential applications in cosmetology. Artemisia ssp. primarily contain sesquiterpenoid lactones, coumarins, flavonoids, and phenolic acids. Essential oils obtained from these species are of great biological importance. Extracts from Artemisia ssp. have been scientifically proven to exhibit, among others, hepatoprotective, neuroprotective, antidepressant, cytotoxic, and digestion-stimulating activities. In addition, their application in cosmetic products is currently the subject of several studies. Essential oils or extracts from different parts of Artemisia ssp. have been characterized by antibacterial, antifungal, and antioxidant activities. Products with Artemisia extracts, essential oils, or individual compounds can be used on skin, hair, and nails. Artemisia products are also used as ingredients in skincare cosmetics, such as creams, shampoos, essences, serums, masks, lotions, and tonics. This review focuses especially on elucidating the importance of the most popular/important species of the Artemisia genus in the cosmetic industry.
... These preparations are indicated for the treatment of inflammation of the colon, rosacea, frostbite, and inflammation of the lymph nodes and mucous membranes [37,43]. Homeopathic medicines from A. abrotanum are also indicated for people suffering from depression and anxiety [63]. The European Medicines Agency (EMA) has also authorized the use of homeopathic preparations from A. abrotanum in farm animals used for food production [11]. ...
... The use of 15, 25 or 40 μL of the essential oil was shown to inhibit the growth of S. aureus, while the use of 10, 15, 25 or 40 μL of the essential oil inhibited the growth of E. coli and C. albicans. The authors of the study indicate that the antimicrobial effect of A. abrotanum can be attributed to the compounds contained in the plant, such as borneol, cymene, camphor, terpineol, 1,8-cineole, and aromadendrene [63]. ...
... The negative control with DMSO and the positive control with methotrexate showed 32.179% and 18.205% survival of RD cells, respectively. The results of the study prove that the essential oil of A. abrotanum, especially in concentrations of 50 and 100 μg/mL, has an antitumour effect, probably due to the presence of compounds such as: borneol, cymene, camphor, terpineol, 1,8-cineole, and aromadendrene [63]. ...
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