Volume 23, Issue 6 (Feb - Mar 2020)                   2020, 23(6): 504-513 | Back to browse issues page

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Moosazad S, Aghaei K, Mahmoudi R, Moosavy S, Shahsavari S. Antibacterial Effects of Kumquat Peel Essential Oil on Staphylococcus Aureus and Escherichia Coli Bacteria in Comparison With Some Standard Antibiotics. Journal of Inflammatory Diseases. 2020; 23 (6) :504-513
URL: http://journal.qums.ac.ir/article-1-2816-en.html
1- Student Research Committee, Qazvin University of Medical Sciences, Qazvin, Iran.
2- Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. , r.mahmodi@yahoo.com
3- Faculty of Medicine, Qazvin University of Medical Sciences, Iran.
4- Department of Epidemiology and Biostatics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
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Extended Abstract
1. Introduction

Foodborne pathogens are a group of microorganisms that cause foodborne diseases. Finding effective drugs against these diseases is essential. Foodborne diseases have increased worldwide and have become a leading cause of death [1]. Several foodborne pathogens have been identified that affect the health and safety of humans and animals. Since the use of antibiotics for the treatment of infectious diseases, it has been observed that bacteria have resistance to them [2]. Due to the increasing desire of consumers to use natural products and food free of chemical preservatives, studies have been conducted on the plants and aromatic fruits and their extracts in terms of preservative properties and the effect on food safety. Their preservative properties are mostly because of their essential oils and metabolite components [3]. 
Kumquat is a fruit-bearing tall tree of the Rutaceae family [4]. Their fruits are eaten whole, and are used in making products such as marmalade and sauces. Dried Kumquat was often used as a traditional medicine for the treatment of respiratory tract infections such as hoarseness and cough [5]. The fruits’ color is orange. This plant is native to China, but today it is grown in many parts of the world, including northern Iran [6]. An important feature of Kumquat plant fruits is their consumable skin which is rich in a variety of metabolites and substances useful for human health [7]. In this study, we aimed to investigate the antimicrobial effect of the essential oil of kumquat peel on foodborne pathogens including Staphylococcus aureus and Escherichia coli in comparison with some standard antibiotics.
2. Materials and Methods
The kumquat fruite was purchased from market in Qazvin, Iran in January 2017. The skin of the fruit was removed with a sharp knife and after crushing, the skin was dried in the shade and at room temperature. After that, the dried skin was thoroughly powdered using an electric grinder machine [8]. The essential oil was extracted by Klevenger apparatus using steam distillation method for 3 hours. For disk diffusion test, the bacteria were first introduced into Brain Heart Infusion Broth (BHI Broth) Then, from colonies in this medium, 0.5 MacFarland was diluted and 100 ppm of pre-prepared suspension was removed and was poured into the Mueller Hinton Agar medium and spread by Pasteur Pipette. Then, paper discs were placed on an empty plate and 20 ppm of different concentrations of essential oil was poured onto them. At the same time, ampicillin, amoxicillin, tetracycline and chloramphenicol antibiotics were used to compare the effect of essential oil. In this regard, the bacteria were cultured uniformly on the Mueller-Hinton agar surface and then placed on the surface of antibiotic-impregnated discs at a specific concentration. Finally, the plates were incubated at 37 °C for 24 hours [9]. Minimum Inhibitory Concentration (MIC) measurements were carried out in a 96-well sterile microplate using broth microdilution technique. First, 100 μl from the BHI Broth medium was poured into the 96- well microplate. To the first well of each row, 100 μl of kumquat peel essential oil (dissolved in dimethyl sulfoxide solvent) was added. Then, 100 μl was taken from the first well and poured into the second well and, after being repeatedly filling and emptying by a sampler, 100μl was taken from the second well and poured into the third well. This work continued until well No. 11. Wells No. 11 of each row, as control essential oil (negative control), contained only medium and essential oil. Wells No. 12 of each row, as control bacteria (positive control) to determine the turbidity of the bacteria, contained the medium, dimethyl sulfoxide and bacteria. To determine the MIC of bacterial growth, the lowest concentration level that had no turbidity, was considered as a MIC level [10]. To measure the Minimum Bactericidal Concentration (MBC), 10 μl were taken from the turbidity-free wells (MIC concentrations and more) under very sterile conditions near the flame, and inoculated and cultured on the surface of Tryptic Soy Agar. After 24-h incubation at 37 °C, the lowest concentration that could kill 99.9% of the bacteria was considered as MBC level [11].
3. Results
The essential oil yield of kumquat peel in this study was obtained 6.66% using following formula 1: 
1. Percentage of essential oil yield=100×(dry weight of plant peel/essential oil weight)
The results of MIC and MBC measurements for essential oil of kumquat peel against the study bacteria are shown in Table 1. The results showed that Staphylococcus aureus is less sensitive to the essential oil compared to Escherichia coli. Based on independent t-test results, the mean levels of MIC and MBC aganist the study bacteria were not significantly different (P=0.089 and 0.468). Their mean levels against the Staphylococcus aureus were 0.42±0.18 and 1.04±0.3, and against the Escherichia coli, they were 0.26±0.09 and 0.42±0.18, respectively. The results of ANOVA showed a significant difference in the diameter of the growth inhibition zones between the five concentrations of extracted essential oil against the study bacteria (P<0.001). In other words, different concentrations of the kumquat peel essential oil had a significant effect on the diameter of the growth inhibition zone.
4. Discussion
Kumquat peel essential oil has significant antibacterial effect which is different depending on the species and the growth region. With high yield of Kumquat fruit, the variety of its species and the increase in its consumption in Iran, the use of this fruit as a combination of antimicrobial and natural preservative, instead of chemical preservatives, in the food and pharmaceutical industries can be beneficial.
Compliance with ethical guidelines
The present study was approved by the Ethics Committee of Qazvin University of Medical Sciences (code: 14003076).
This study was extracted from the research proposal of first author (Code: 14003076) supported by the Research Committee of Qazvin University of Medical Sciences.
Authors' contributions
Writing and investigation: Sara Moosazad; Methodology and experiments: Katayoon Aghaei, Shaghayegh Moosavy; Supervision, project administration, and funding acquisition: Razzagh Mahmoudi; Data curation and analysis: Saeed Shahsavari.
Conflicts of interest
 The authors declared no conflict of interest.
Authors appreciate Deputy of the Vice Chancellor for Research in Medical Sciences of Qazvin University.
Type of Study: Research | Subject: Microbiology

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