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\title{Argan Oil as A Novel Anti-Methicillin Resistance}
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             \author[1]{Dr. Hussein O. M. Al-Dahmoshi Al-Enezi  }

             \affil[1]{  Babylon University, College of Medicine, Microbiology Dept.}

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\date{\small \em Received: 10 December 2013 Accepted: 31 December 2013 Published: 15 January 2014}

\maketitle


\begin{abstract}
        


Methicillin-resistant Staphylococcusaureus (MRSA) infections are most medically threat, hard to treat infection among medical care units. MRSA isolates emerged from Staphylococcusaureus strain how carry mecA gene which confer their resistance to methicillin. The standard drug of choice to resolve MRSA infections is vancomycin which late replaced by teicoplanin due to their renal toxicity. During this study 20 MRSA isolates were obtained from the Central Laboratory of Babylon Health Directorate and previously diagnosed as MRSA using both phenotypic (by VITEK 2 compact system) and genotypic (mecA gene detection) diagnostics methods. Teicoplanin disk diffusion were achieved acc -ording to CLSI 2012.Well diffusion method was used to test the effects of mixture of Argan Oil and 1.5 % H2O2 with ratios: (1:1, 1:2, and 2:1 Argan oil: 1.5% H2O2). The effect of the mentioned mixtures in addition to the effect of 1.5% H2O2 alone and Argan oil alone were tested using well diffusion method according to CLSI 2012. The results of inhibition zone for mixture were recorded and compared with the inhibition of teicoplanin. The results revealed ability of argan oil: H2O2 mixture with ration 2:1 to inhibit growth 80% of MRSA isolates and give inhibition zone similar to those of teicoplanin ? 14mm. This study concludes the satisfactoriness of use Argan oil as anti-MRSA medical oil with high level of safety and low level of uncomfortable effects.

\end{abstract}


\keywords{argan oil, mrsa, teicoplanin, tocopherol.}

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\let\tabcellsep& 	 	 		 
\section[{I. Introduction}]{I. Introduction}\par
ethicillin-resistant Staphy Lococcus Aureus (MR -SA) is a bacterium accountable for several hard to treat infections among human. Any isolate of S. aureus that become resistant to ? -lactam antibiotics, like penicillins (methicillin, dicloxacillin, nafcillin and oxa -cillin) and cephalosporins, called MRSA; while those isolates that incapable to resist these antibiotics are classified as methicillin-sensitive Staphy Lococcus Aureus, or MSSA [1]. Staphy Lococcus Aureus is one of the most frequent bacterial causes of MRSA is particularlyworrying in hospitals and nursing homes, where patients with open wounds, invasive de -vices, and weakened immune systems are at greater risk of infection than the general public \hyperref[b1]{[2]}. 
\section[{Staphy Lococcus Aureus mainly colonize}]{Staphy Lococcus Aureus mainly colonize}\par
nostrils; open wounds, intravenous catheters, and the urinary tract are also potential sites for infection. MRSA are often classified as Hospital-Associated MRSA (HA-MRSA) or Community-Associated MRSA (CA-MRSA), dependent upon the situations of disease acquisition \hyperref[b3]{[3]}.\par
Medical oils, essential oils and Plant extracts have importance to use for different purposes. Essential oils have been searched for their antibacterial, antifungal, antiviral, insecticidal, anticancer and antioxidant properties \hyperref[b4]{[4]}. All these plant oils were proved to have antibacterial activity which is mainly attributed to the presence of two plant phenolic compounds, thymol and carvacrol \hyperref[b5]{[5]}. The anti-MRSA effects of many essential oils like thymus and eucalyptus oils were studied and the results revealed high sensitivity of MRSA to these oils using both disk and well diffusion methods and these effects attributed to the presence of thymol and to eucalyptol in thymus and eucalyptus respectively \hyperref[b6]{[6]}.\par
Argan oil is a plant oil produced from the kernels of the Argan tree (Argania spinosa L.) that is endemic to Morocco. The fruits of the Argan tree are nut-sizes and may be round, oval or conical in shape. The fruits are covered by a thick peel which covers the fleshy pulp. The pulp surrounds a hard-shelled nut which repr-esents approximately 25\% of the weight of the fresh fruit. Contained within the nut are one to three Argan oil-rich kernels. Argan oil is extracted from the kernels, with yields varying from 30\% to 55\% depending on the extraction method used \hyperref[b8]{[7]}. Argan oil consist of 42.8\% oleic, 36.8\% linoleic, 12\% palmitic, 6\% stearic and <0.5\% Linolenic. Argan oil contains tocopherols (vitamin E), phenols, carotenes, squalene, and fatty acids, (80\% unsaturated fatty acids). The main natural phenols in  The percentage of sensitivity of MRSA isolates were 0.00\%, 0.00\%, 45.00\%, 80.00\% and 70.00\% for (Argan oil, 1.5\% H2O2, 1:1, 2:1 and 1:2 ratio of Argan oil:H2O2 respectively) (figure2). All MRSA isolates show no sensitivity to argan oil alone and to H2O2 alone while 45\%, 80\% and 70\% of isolates were sensitive to 1:1, 2:1 and 1:2 of Argan oil: H2O2 respectively.  
\section[{Mixture Ratio}]{Mixture Ratio} 
\section[{Mean of Inhibition Zone (mm)}]{Mean of Inhibition Zone (mm)}\par
argan oil are caffeic acid, oleuropein, vanillic acid, tyrosol, catechol, resorcinol, epicatechin and catechin.\par
Depending on the extraction method, argan oil may be more resistant to oxidation than olive oil \hyperref[b9]{[8]}.\par
Until yet few studies concern antibacterial effect of Argan oil published. The positive antipseudomonal effect of Argan oil against pseudomonasaeruginosa isolated from burn was stated in Iraq previously \hyperref[b10]{[9]}. While the antibacterial effect of argan oil against MRSA not stated otherwise, the current study aimed to study the effect of argan oil against MRSA using well diffusion method in comparison with first choice antibiotics to get rid MRSA infection, Teicoplanin. 
\section[{II. Material and Methods}]{II. Material and Methods} 
\section[{a) Bacterial Isolates}]{a) Bacterial Isolates}\par
Twenty MRSA isolates were used in this study. All isolates obtained from the Central Laboratory of Babylon Health Directorate and previously diagnosed as MRSA using both phenotypic and genotypic diagnostics methods. For more confirmation all isolates were rediagnosed using VITEK 2 compact system. 
\section[{b) Well Diffusion Method}]{b) Well Diffusion Method}\par
The virgin stock argan oils bring from Morocco and the mixture of Argan Oil and 1.5 \% H2O2 were prepared with following ratios: (1:1, 1:2, and 2:1 Argan oil:1.5\% H2O2). The effect of the mentioned mixtures in addition to the effect of 1.5\% H2O2 alone and argan oil alone were tested using well diffusion method. According to CLSI 2012 \hyperref[b11]{[10]}, the MRSA isolate suspension was standardized with 0.5 McFarland and the streaked on Muller Hinton agar plates and the wells made using cork poorer after inoculation of each plate with MRSA isolate. Each plate contains five wells for 1:1, 1:2, and 2:1 Argan oil: 1.5\% H2O2 I addition to 1.5\% H2O2 alone and Argan oil alone. The results recorder as diameter of inhibition zone and then comparing the results with the diameter of inhibition zone of commonly used antibiotic for Staphy Lococcus Aureus treatment. 
\section[{c) Disk diffusion Method}]{c) Disk diffusion Method}\par
This method used to check the sensitivity of all MRSA isolates to teicoplanin according to CLSI 2012. 
\section[{III. Results}]{III. Results}\par
The effects of Argan oil on MRSA not studied in Iraq and most of Arabian countries yet. Effect of this oil on MRSA isolates were recorded as inhibition zone (mm) and the results revealed no effect of 1.5\% H2O2 alone and Argan oil alone. The effect of Argan oil: An H2O2 mixture was positive and gives same results using different ratio (1:1, 1:2 and 2:1 of Argan oil: H2O2). The mean of inhibition zone were 13.2mm, 14mm and 14.4mm respectively (figure \hyperref[fig_2]{1}). To increase the acceptability of Argan oil as anti-MRSA, the comparison of their effect with common antibiotics used to treat MRSA infections must be achieved. Both teicoplanin and vancomycin are effective against MRSA,but there is an apprehension that vancomycin may be more toxic, especially for the kidneys and so the comparison is with effect of teicop -lanin and argan oil. The results display that all isolates (100\%) were sensitive to (inhibition zone ?14mm) while 6(30\%), 16(80\%) and 11(55\%) of isolates were sensitive (give same results for teicoplanin (inhibition zone ?14mm)) to 1:1, 2:1 and 1:2 of Argan oil: H2O2 respectively table \hyperref[b0]{(1)}. The results show the similar effects of argan oil and teicoplanin especially for 2:1 Argan oil: H2O2 (high argan oil concentration mixture).  
\section[{Volume XIV Issue III Version}]{Volume XIV Issue III Version} 
\section[{IV. Discussion}]{IV. Discussion}\par
The central dogmas of using Argan oil as anti-MRSA agents that, herbal oil with great medical value, safe, cheap and available. The real needs to use medical or herbal oil emerged from the fact that there is little or no resistance along with reducing their disadvantageous side effects of the routine antibiotics \hyperref[b12]{[11]}. There is no value for H2O2 as anti-MRSA and be used as solubilizer to facilitate dissemination and dispersion of Argan oil through the Muller Hinton agar plate and have the same effect as tween 20 \hyperref[b6]{[6]}.\par
The glycopeptides antibiotics vancomycin and teicoplanin remain the standards for treating most MRSA infections but the toxicity of vancomycin push the teicoplanin to be first choice against MRSA infections and the teicoplanin wassuperior in terms of antibacterial effects \hyperref[b13]{[12]}. the great findings that the argan oil has similar in vitro effect of teicoplanin and can be used as safely alternative medication to get rid such infections.\par
Argan oil consists of 42.8\% oleic acid, 36.8\% linoleic acid, 12\% palmitic acid, 6\% stearic acid and 0.5\% Linolenic acid. It is also contain Unsaponifiable matter,Such as carotenes, tocopherols, triterpene alcohols, sterols, and xanthophylls. Specific health benefits of argan oils are attributed to its composition of unsaponifiable matter and high tocopherol content \hyperref[b14]{[13]}.Gamma-Tocopherol composes 69\% of Argan oil total tocopherol content. Because tocopherols and sterols can act synergistically, the specific combination of molecules found in the unsaponifiable matter is theorized to contribute to the therapeutic aspects of Argan oil \hyperref[b15]{[14]}. Previous studied revealed great effect of Argan oil on Pseudomonasaeruginosa isolates in Iraq \hyperref[b10]{[9]}. 
\section[{V. Conclusion}]{V. Conclusion}\par
Current study reveal the beneficially effect of argan oil to treat MRSA infections beside their safety for human use and accessibility in addition to their role as antioxidants. microenvironments and interspecific interactions influence nasal microbiota complexity and S.aureus carriage". Cell Host Microbe 14 (6): 631-40.\begin{figure}[htbp]
\noindent\textbf{}\includegraphics[]{image-2.png}
\caption{\label{fig_0}}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{}\includegraphics[]{image-3.png}
\caption{\label{fig_1}}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{1}\includegraphics[]{image-4.png}
\caption{\label{fig_2}Figure 1 :}\end{figure}
  \begin{figure}[htbp]
\noindent\textbf{1} \par 
\begin{longtable}{P{0.4407407407407407\textwidth}P{0.20148148148148148\textwidth}P{0.1385185185185185\textwidth}P{0.06925925925925926\textwidth}}
Agent\tabcellsep Inhibition zone (mm)\tabcellsep Sensitivity No.\tabcellsep \%\\
Teicoplanin\tabcellsep ?14\tabcellsep 20\tabcellsep 100\\
1:1 Argan oil:H2O2\tabcellsep ?14\tabcellsep 6\tabcellsep 30\\
2:1 Argan oil:H2O2\tabcellsep ?14\tabcellsep 16\tabcellsep 80\\
1:2 Argan oil:H2O2\tabcellsep ?14\tabcellsep 11\tabcellsep 55\end{longtable} \par
 
\caption{\label{tab_0}Table 1 :}\end{figure}
 			\footnote{Argan Oil as A Novel Anti-Methicillin Resistance Staphylococcus Aureus (MRSA), Iraq} 		 		\backmatter  			  				\begin{bibitemlist}{1}
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\end{document}
