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African Journal of Agricultural Marketing ISSN 2375-1061 Vol. 8 (9), pp. 001-004, September, 2020. Available online 
at www.internationalscholarsjournals.org © International Scholars Journals 

 

Author(s) retain the copyright of this article. 
 
 
 
 

Full Length Research Paper 

 

Antimicrobial activity evaluation of the oleoresin 

oil of Pistacia vera L. 

 
Bachir Raho Ghalem* and Benali Mohamed 

 
Faculty of Biotoxicology Laboratory Science, Institute of Biology, Djillali Liabès University of Sidi Bel Abbès, Algeria. 

 
Accepted 05 May, 2020 

 
The hydrodistilled essential oils from the exudates of Pistacia vera L. stems have been tested against 

three bacteria, Escherichia coli, Staphylococcus aureus and Proteus using three methods: agar disc 
diffusion method, determination of MIC (minimal inhibitory concentration) and in the liquid phase, by 
Maruzella method. The results obtained showed that E. coli was the greatest inhibitor of all the strains 

tested, and that Gram negative bacteria (E. coli and Proteus spp.) showed more inhibition than that 
observed on Gram positive bacteria (S. aureus) by the essential oil tested. 

 
Key words: Essential oils, oleoresin of Pistacia vera L., strains tested, antimicrobial activity determination. 

 
 
INTRODUCTION 

 
Among the aromatic plants belonging to the family of 
Anacardiaceae, the genus Pistacia is noteworthy for its 
numerous species and varieties of wild-growing plants. 
Many of these species are typical of the Mediterranean 
area. Pistacia has an economic value as it is the source 
of traditional medicinal agent "gum" mastic, an oleoresin 
exudates from the stem of this plant (Dogan et al., 2003). 
It is a traditional natural remedy used in very ancient 
civilizations in the Mediterranean like Greek and Egyptian 
(Pellecuer et al., 1980; Langenheim, 2003; Peachey, 
1995). In Algeria, it is found in four species, namely 
Pistacia lentiscus, Pistacia terebinthus, Pistacia atlantica 
and Pistacia vera. According to ecology, the true 
pistachio (P. vera) is characterized by a large tolerance to 
climatic variations; it can grow under slices rainfall quite 
low and can cope in soils. In Algerian folk medicine, 
Pistacia has been used as an astringent, expectorant and 
cicatrisant agent (Benhammou et al., 2008). 

About the other genus of this plant, investigations have 

shown some pharmacological effects such as reducing 

blood pressure (Villar and Paya, 1987), anti- inflammatory  
 
 

 
*Corresponding author. E-mail:bachir_raho@yahoo.fr. Tel: 
00213771063841. 

 
 
 
 

 
(Giner et al., 2001; Giner et al., 2000) and antimicrobial 

action (Ali-Shtayeh and Abu, 1999; Magiatis et al., 1999). The 

antiseptic activity of P. lentiscus essential oils and its resin on 

different microorganisms has been reported by several 

researchers (Tassou and Nychas, 1995; Iauk et al., 1996; 

Ali-Shtayeh and Abu, 1999; Marone et al., 2001; 

Benhammou et al., 2008; Douissa et al., 2005) but the 

antimicrobial effect of Pistacia vera extracts precisely its 

oleoresin oils have not been studied so far (Duru et al., 

2003; Kordali et al., 2003; Özçelika et al., 2005) . In this 

study, we aimed to detect the inhibitory effect of the oils 

extracts from exudates of oleoresin from the P. vera stem on 

the growth of Escherichia coli, Staphylococcus aureus and 

Proteus tested by using three methods: agar disc diffusion 

method, determination of MIC (minimal inhibitory 

concentrations) and in the liquid phase, by Maruzella 

method. 
 
 
MATERIALS AND METHODS 
 
Plant material and extraction of the essential oil 
 
The essential oil of P. vera was extracted from the stem oleoresin 
by water distillation. The mastic gum was collected from the 

Technological Institute of fruit trees –T.I.F.T- of Tighennif (Wilaya of 
Mascara) situated in the northwest of Algeria, in the months of April, 

file:///C:\Users\user\Documents\REPUBLICATION\AGRICULTURAL%20SCIENCES\AppData\Local\Temp\www.internationalscholarsjournals.org


  
 
 

 
Table 1. Antimicrobial activity evaluation of the essential oil resin of P. vera with agar disc diffusion method.  

 

Bacterial strain 
    Resin concentration (µg/ml)    

 

 

Dilution 10
-1

 10
-2

 10
-3

 
  

10
-4

 
 

Standard 
 

     
 

 10
-1

 10 9 8.5   7 7 
 

E.coli 10-2 11 10.5 9   8.5 7 
 

 10
-3

 11.5 10.9 9.5   9 6.5 
 

 10
-1

 9 8.5 8   7 6.5 
 

S.aureus 10
-2

 10 9 8.5   7.5 7 
 

 10
-3

 11 10.5 10   8.5 6.5 
 

 10
-1

 9 8 8   7.5 7 
 

Proteus spp. 10
-2

 11 10.5 10.5   9 7.5 
 

  10
-3

 11.5  11.5  11.5   10.5  7 
 

 
 

 
May and June, which corresponds to the period of the oleoresin 

formation. 

 

Bacterial strains 
 
All bacterial strains (E. coli, Proteus spp and S. aureus) were 
provided by the Laboratory of Medical Analysis-–located in Dr. 
Yessaâd Khaled Hospital (YKH) of Mascara City, situated in the 
west of Algeria for patients suffering from certain infectious 
diseases. S. aureus was isolated from the pus of a patient, E. coli 
from blood specimens while Proteus was taken from a coproculture, 
and then confirmed by biochemical tests and morphological studies 
in Microbiology Laboratory of Biology Institute in the Mascara 
University (Euzéby, 1998; Marchal et al., 1982). 

 

Antimicrobial activity determination 
 
We used three methods to determine the antibacterial activity; agar 
disc diffusion method, determination of MIC (Minimal Inhibition 
Concentrations) and in the liquid phase, by Maruzella method. The 
agar disc diffusion method was employed to determine the anti-
microbial activities of the essential oils in question. A suspension of 

each sample tested micro organism - diluted prior to 10
-1

, 10
-2

 and 

10
-3

 - (1 ml of 10
8
 cells/ml), was spread on the solid media plates. 

Filter paper discs (6 mm in diameter) were soaked in 13 µl of the 
resin oil and placed on the inoculated plates and, after drying for 15 
min, were incubated at 37°C for 24 h .The diameters of the 
inhibition zones were measured in millimetres (Tepe et al., 2004).  

Minimal inhibitory concentration (MIC) was taken from the 

concentration of the lowest dosed test tube showing visually no growth. 

10 µl from each visually no grown test tube was subcultured on Mueller-
Hinton agar (Pauli and Kubeczka, 1996) . Each strain from the three S. 

aureus, Proteus spp and E. coli was diluted at 10
-1

, 10
-2

 and 10
-3

 (1 ml 

of 10
8
 cells/ml); a standard was prepared with ethanol and oleoresin oil 

of P. vera. From each dilution a slick strain was spread over the surface 

of the Petri dish containing Mueller-Hinton agar medium liquid which 
was dried at 37°C for 15 min. Four discs were placed on agar containing 

the following quantities of the oleoresin oil dilution: 0.5, 1, 1.5, 2 and 2.5 
l. Ethanol was added to the standard disc, placed in the Petri dish 

center, and was incubated toward the end at 37°C for 24 h. The 
principal technique of Maruzella method is to act in the liquid phase of 

increasing concentrations of oleoresin essential oil, after emulsifier 

addition 

 

 

(Singh et al., 2000; Larrondo et al., 1995). Serial dilutions (10
-1

, 10
-
 

2 and 10
-3

 µml) were prepared from this essential oil solution. 1 ml 
of each dilution and 0.5 ml of tested culture strains were added to 8 
ml of nutrient broth, maintained in a Bain Marie to 37°C under 
agitation for 24 h and then seeded by streaking the surface of agar 
medium and incubated at 37°C for 24 h.

 

 

 

RESULTS AND DISCUSSION 

 

As indicated in Table 1, all strain bacteria were inhibited 
by essential oil resin of P. vera. A more significant 
inhibition was seen with a higher oleoresin oil con-
centration. At low concentrations, a very limited inhibitory 
effect was observed on the growth of microorganisms in 
comparison with those of the standard. With increasing 
essential oil resin of P. vera concentration, an obvious 
inhibitory effect on growth of, E. coli, Proteus spp and S. 
aureus was significantly increased.  

The addition of mastic gum oil in broth culture 
inoculated with S. aureus, E. coli and Proteus spp inhi-
bited the growth of these organisms .The rate of inhibition 
was greater, on Gram negative bacteria (E.coli , Proteus 
spp), than that observed on Gram positive bacteria (S. 
aureus). In most cases the size of inoculum and the con-
centration of mastic gum oil affect the growth/survival of 
the organisms. These results are almost similar to those 
shown in other work on the antimicrobial activity of oil 
mastic gum of P. vera as well as those of similar species 
(Iauk et al., 1996; Koutsoudaki et al., 2005; Kamrani et 
al., 2007; Benhammou et al., 2008 and Özçelik et al., 
2005). Minimal inhibitory concentration (MIC) values were 
defined as the lowest concentration of oils that completely 
inhibited microbial growth. The results were expressed in 
micrograms per millilitre. The results for the MIC are 
shown in Table 2. The MIC values regarding the 
antimicrobial activity of oil mastic gum of P. vera against 
Gram negative bacteria (E. coli and Proteus spp.) and S. 
aureus (Gram positive bacteria) were determined to be 



 
 
 

 
Table 2. MIC evaluation of the essential oil resin of P. atlantica.  

 

Bacterial strain 
     Resin concentration (µg/ml)    

 

 

Dilution 0.5 1 1.5 2 
  

2.5 
 

Standard 
 

     
 

 10
-1

 6.5 6.5 9 9.5   10 6 
 

E.coli 10
-2

 7 7 9.5 10   11 6.5 
 

 10
-3

 6.5 8 10 10.5   11 7 
 

 10
-1

 6 7 8 10   11 6 
 

S. aureus 10
-2

 6.5 7 9 10   11.5 6.5 
 

 10
-3

 6.5 7.5 9.5 10.5   12 6.5 
 

 10
-1

 6.5 8 9 10   10.5 6 
 

Proteus spp. 10
-2

 7 9 9.5 11   11 6 
 

  10
-3

  7  8.5 10.5  11.5   12  6.5 
 

 

 
Table 3. MIC evaluation essential oil resin of P. atlantica with the three bacterial strains.  

 
 

Bacterial strain 
 Essential oil (µg/ml)  

 

 

Standard «0» 10
-1

 ,10
-2

 10
-3

, 10
-4

 10
-5

  

  
 

 E.coli + ++ + ++ +++ 
 

 S. aureus +++ + ++ +++ 
 

 Proteus spp +++ + +++ +++ 
 

 
++: Comparable growth with that witness. 
+: Slow growth. 

 

 

1.5 and 2.0 µg/ml, respectively. The results indicated that 

the oil mastic gum of P. vera showed antibacterial activity, 

according to Alma et al. (2004), and Özçelik et al. (2005), 

mainly against the Gram-negative bacteria (E. coli and 
Proteus spp). The oil mastic gum also exhibited an effect 

against the Gram-positive bacteria (S. aureus). However, this 

effect was less efficient than that presented against the Gram-

negative bacteria, since a higher MIC value was obtained with 

the Gram-positive bacteria. Differences in MIC values of 

bacteria may be related to differential susceptibility  
of bacterial cell wall, which is the functional barrier to  
minor differences present in outer membrane in the cell wall 

composition (Zhao et al., 2001). Like previous tests, the 

application of the liquid phase method confirmed( by its results 

showed in Table 3) the important antibacterial activity of the oil 

mastic gum of P. vera on these three microbial strains, as it 

seems that Proteus spp is more sensitive than the other two. 

The change of inhibitory effect depends on the natural 

substance concentration. 

 

Conclusion 
 
The results of the antimicrobial activity tests indicate that 

 
 

 

essential oil of mastic gum P. Vera exhibited higher 

activity against the tested strains and confirm its 
traditional uses. However, oil mastic gum was found to 
inhibit both gram-positive and gram-negative bacteria. 
We believe that the present investigation together with 
previous studies provide a support to the antibacterial 
properties of this essential oil. It can be used as an 
antibacterial supplement in developing countries towards 
the development of new therapeutic agents to treat 
several infectious diseases caused by these pathogens. 
 

 
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