Submitted: 26 Feb 2017
Revised: 07 May 2017
Accepted: 17 May 2017
First published online: 20 May 2017
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Antibacterial Properties of Essential Oil of <i>Heracleum persicum</i> (Golpar) and Foodborne Pathogens

Int J Enteric Pathog, 5(2), 41-44; DOI:10.15171/ijep.2017.10

Original Article

Antibacterial Properties of Essential Oil of Heracleum persicum (Golpar) and Foodborne Pathogens

Nabi shariatifar1, Toktam Mostaghim2, Afshin Afshar3, Issa Mohammadpourfard4 ,*, Mehran Sayadi5, Mohammad Rezaei4

1 Department of Environmental Health, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
2 Department of Food Science and Technology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran
3 Departement of Food and Drug, Alborz University of Medical Sciences, Karaj, Iran
4 Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
5 Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

*Corresponding Author: Issa Mohammadpourfard, Tel: +989149080532; Email: Isamohammadpour@yahoo.com

Abstract

Background: Antimicrobial resistance is a serious health threat to human society. Antibiotics and preservatives are also the most common modalities to increase the shelf life of foods.
Objectives: Antibacterial activity of essential oil of Heracleum persicum (Golpar) fruit against some of the main foodborne bacteria was determined.
Materials and Methods: Antibacterial activity of essential oil was evaluated against the bacteria (Staphylococcus aureus, Escherichia coli, Salmonella typhi, Vibrio cholera, and Yersinia enterocolitica) using disc diffusion method. Broth micro-dilution method was used to determine their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The data was expressed as mean and standard deviation and analyzed using analysis of variance (ANOVA) in SPSS software (P < 0.05).
Results: Comparing with S. typhi strains (inhibition zone = 18 ± 0.46 mm), essential oil was found to be more effective against V. cholera strains (inhibition zone = 34 ± 0.2 mm). The MIC value (%) of the essential oil against V. cholera and S. typhi were 8 and 32 respectively. Essential oil of H. persicum (Golpar) showed a good antimicrobial activity against foodborne pathogens.
Conclusion: The results revealed that the essential oil of H. persicum can be used in food preservation systems to inhibit the growth of V. cholera strains and improve food quality and safety.

Keywords: Antimicrobial activity, Essential oil, Food Quality, Heracleum persicum, Pathogenic bacteria

Copyright © 2017 The Author(s); Published by Alborz University of Medical Sciences. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

Background

Foodborne pathogens belong to a group of micro-organisms that cause foodborne diseases and it is essential to find effective drugs against the infection.1 Prevalence of foodborne diseases has increased worldwide and become a major cause of mortality in individuals with impaired immune systems in the developing countries.2 There is an urgent need to monitor antimicrobial resistance by improving the ways the antibiotics may be used and reduce hospital cross-infection. New antibiotics need to be developed continuously to maintain the effectiveness of antimicrobial therapy.3-5 The recent decades have demonstrated an increase in interests in the use of herbal products.6 Plants play an important role in human health. It is estimated that 25% of modern medicines are derived directly or indirectly from herbs.7,8 Many plant compounds have been proven to be effective insecticidal, antifungal, anti-bacterial, anti-viral, anti-parasitic, and anti-oxidant agents.3,9 Aromatic essential oils are often used in food industry to add flavor and prevent growth of microbial and mold contaminants.10Heracleum, which is used for medicinal purposes, is an annual herbaceous plant. It is a genus of about 60 species, which mainly grows in Iran and Europe.11,12Heracleum persicum is known as Heracleum in Farsi and as tromso palm in Norwegian. Fruits of this plant are used as spices and as anti-infection and analgesic in traditional medicine and the stems are used in pickles.13 Many studies have been conducted on the antibacterial effect of Heracleum and most of them have been on the leaves and stems; still, there has been no study on antibacterial effect of essential oil of its fruits. Literature review showed a paucity of studies on the antibacterial effects of chemical composition of the oil of H. persicum fruit on foodborne pathogens.

Objectives

This study aimed at determining the antibacterial effects of essential oil of Heracleum fruit on some of the foodborne pathogens.

Materials and Methods

Plant Materials

Heracleum fruits were collected in May 2013 from the heights in Shemiranat-Tehran and then identified and confirmed in Herbarium Center of School of Pharmacy, Tehran University of Medical Sciences. The collected plants were dried in the shade and then the fruits were separated from the stems and ground by mechanical means.

Chemicals and Reagents

All chemicals and standards were obtained from Merck (Darmstadt, Germany).

Extraction of Essential Oil

The essential oils of the Heracleum were obtained using hydro distillation and a Clevenger apparatus; 300 g of fruits was placed in the Clevenger with sufficient distilled water to cover the material. Extraction continued for 3 consecutive hours when the water started to boil.14

Antimicrobial Activity Test

Disk Diffusion Test

The essential oils were tested against Staphylococcus aureus ATCC 25913, Escherichia coli ATCC 8739, Salmonella enterica PTCC 1709, Vibrio cholera PTCC 1611 and Yersinia enterocolitica PTCC 1477. The microorganisms were cultured in BHI (brain heart infusion) for 18 hours at 37ºC, compared with 0.5 McFarland standard (5 × 108 CFU/mL), and inoculated directly in boards with Müller-Hinton agar (Merck). After inoculation of each microorganism, the diffusion was carried out; so that 10 μL of the essential oil was placed on paper discs (6 mm of diameter) at 37ºC for 24 hours. Afterward, the zone of inhibition was measured.3

Minimum Inhibitory Concentration and Minimum Bactericide Concentration

To obtain minimum inhibitory concentration (MIC) and minimum bactericide concentration (MBC), quantitative tests were performed using 1%-9% concentrations of the essential oil in Müller-Hinton broth medium. The concentrations were added by 1×106 CFU/mL of the bacteria. Positive control (cultured along with the bacterium without the essential oil) and negative control (cultured without the bacterium) were also prepared. Then, the samples were placed in an incubator (37ºC, 24 hours). Before reading the results, the last concentration without opaqueness (no growth) was adopted as MIC for each sample. The samples without turbidity were cultured in Müller-Hinton broth medium. After incubation (24 hours, 37ºC), the last concentration suitable enough to be used as bactericide was adopted as MBC.

Statistical Analysis

Each experiment, from sample preparation to analysis, was repeated for three times, and the data were analyzed using one-way analysis of variance (ANOVA) and Tukey tests in SPSS (v. 16).

Results

The results of disc diffusion method showed antimicrobial activity of essential oil of H. persicum against all of the bacterial strains; including S. aureus (inhibitory zone: 32.16 ± 0.76 mm), S. enterica (inhibitory zone: 18 ± 0.46 mm), E. coli (inhibitory zone: 19 ± 1.32 mm), V. cholera (inhibitory zone: 34 ± 2 mm), and Y. enterocolitica (inhibitory zone: 22.16 ± 0.76 mm) (Table 1).

Table 1. Average Inhibitory Halo Diameter (mm) of Various Bacterial Strains for Heracleum persicum Essential Oil
Bacterial Strain Samples Range Mean ± SD
Staphylococcus aureus ATCC 25913 3 32.5-33 32.16 ± 0.76a
Escherichia coliATCC 8739 3 17.5-20 19 ± 1.32b
Salmonella enterica PTCC 1709 3 18-19.5 18 ± 0.46b
Vibrio choleraPTCC 1611 3 32.5-34 34 ± 2a
Yersinia enterocolitica PTCC 1477 3 22-22.5 22.16 ± 0.76a

The results showed that yield of essential oil of Heracleum fruit was about 11%. Impact of the essential oil on each bacterium (Staphylococcus, E. coli, Yersinia, Vibrio, and Salmonella) is listed in Table1. The highest and lowest zones of growth inhibition at concentration of 100% were observed in Vibrio and in Salmonella respectively. In addition, minimum and maximum MIC against bacteria were in‏ Vibrio and Yersinia respectively. As listed in Table 2, Yersinia and Salmonella (64%) and Staphylococcus (16%) have the highest and lowest values of MBC. There was no significant difference (P < 0.05) between the bacterial strains in terms of inhibitory halo diameter‏ except for‏ E. coli and Salmonella.

Table 2. MIC and MBC Value (%) for Heracleum persicum Essential Oil Against Pathogenic Bacterial Strains
Staphylococcus aureus Vibrio cholera Yersinia enterocolitica Escherichia coli Salmonella
MIC 11 8 18 30 32
MBC 16 16 64 32 64

Discussion

Many antimicrobial drugs have been found and there are still more drugs in the nature not found yet. In light of this, the past decade has witnessed an increase in research on plants as medicine source.15,16

Essential oils are potential sources of novel antimicrobial compounds especially against bacterial pathogens. Essential oil plants may also be applied for the treatment of infectious diseases, thanks to its antimicrobial activity.15,16 Results from in vitro studies in this paper showed that the essential oils can inhibit bacterial growth; although, their effectiveness varied.

Nazemi et al indicated that aqueous extract of Heracleum did not have any antibacterial effect; however, Heracleum methanol extract had significant effect on the bacteria Bacillus polymyxa, Bacillus subtilis, Enterococcus faecalis, Nocardia, and S. aureus.17,18

Kousha and Bayat indicated that extracts of leaves and flowers of H. persicum had antimicrobial activity against 4 gram-negative and 4 gram-positive bacteria.19

Manikandan et al20 reported that the methanolic extract of leaves of different plants were more active than the aqueous extract in presence of Bacillus subtilis, Bacillus pumilus, Micrococcus luteus and S. aureus.

Habibi et al21 showed that the highest inhibitory effect of H. rechingeri species was against gram-positive bacteria and B. subtilis in particular. Consistent results were obtained at concentration level of 50% (P < 0.05).

Hemati et al indicated that essential oil and hydro-alcoholic extract of H. persicum had analgesic, anti-inflammatory, and antioxidant effects.22

Inhibition effects of H. persicum (57.16%) on Agrobacterium tumefaciens was reported by Dehghan Noudeh et al.23 They reported that H. persicum (57.16%) had inhibiting effect on all tested gram-positive and gram-negative strains. Our results were consistent with the results of that study.

Frey and Meyers24 indicated that the traditional medicines were capable of creating inhibition zone against the bacteria S. typhimurium, S. aurous, E. coli and Streptococcus lactis. Moreover, their data supported the hypothesis that the selection and use of these plants for disease treatment was not random.

Comparison of our results with those obtained by Dehghan Nodeh et al indicated that 4 fractions from Heracleum had antimicrobial activity against A .tumefaciens, E. coli, S. aureus, B. subtilis and P. aeruginosa and the highest and lowest MIC were observed in E. coli and B. subtilis respectively.23

Several studies have been conducted on antibacterial activity of herbs, which are rich in monoterpenoids. Using drop diffusion method, these studies have shown significant inhibition zones for all microorganisms under study such as gram-positive bacteria (S. aureus) and fungus (Candida albicans).25,26 Results of the present study were consistent with other studies as to gram-positive (S. aureus) and gram-negative bacteria (E. coli, S. enterica, V. cholera, and Y. enterocolitica).

Inconsistencies between the present study and other works might be because of different genotypes of H. persicum utilized, growth ecosystem (habitat, temperature, height), and setting of the experiments (pH and temperature). Several factors influence growth and performance of the plants in an ecosystem including type, habitat, soil, height, and geographical position. Some of these factors have notable effects on the results quantitatively and qualitatively.

Medical herbs as immunostimulants have been widely used as food additives for years and the procedures have been recorded by several research works. Moreover, in vivo, in vitro, and ex vivo surveys of the effects of Heracleum sp. used as an immunostimulant agent in terrestrial and aquatic animals or human require deeper investigations. In addition, more studies are needed on antiviral effects of this family.

Given the information obtained from these studies and the present study, anti-bacterial properties of Heracleum against the pathogens transmitted by food is supported.

Conclusion

Notable antimicrobial activity against foodborne pathogenic bacteria was demonstrated by the essential oil of Heracleum. The essential oil may be effective on other gram-positive and gram-negative bacteria. The practical application of essential oils is to increase shelf life of food. Therefore, it can be a good alternative for artificial preservatives used in the food industry today.

Authors’ Contributions

This article was extracted from the MSc project of IM where NS supervised these project and suggested the problem; TM, AA and MS participated in determination of sample points, sample preparation procedure and extraction of essential oil; MR wrote and edited the manuscript. All authors read and approved the final manuscript.

Ethical Approval

This study was approved by Ethics Committee of Tehran University of Medical Sciences.

Conflict of Interest Disclosures

The authors declare that they have no conflict of interests.

Financial Support

This study was supported by Research Committee of Tehran University of Medical Sciences.

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