Antibacterial activities of Cuminum cyminum Linn. Essential Oil Against Multi-Drug resistant Escherichia coli

AUTHORS

Mohammad Bokaeian 1 , Yasub Shiri 2 , Saphora Bazi 3 , Saeide Saeidi ORCID 4 , * , Zahra Sahi 4

1 Infectious Disease and Tropical Medicine Research Center, Zahedan University of Medical Scienes, Zahedan, IR Iran

2 Department of Agronomy and plant Breeding, Agriculture Research Center, Zabol University, Zabol, IR Iran

3 Department of Biology, Faculty of Sciences, Payam nour University, Zabol, IR Iran

4 Department of Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University, Kerman, IR Iran

How to Cite: Bokaeian M, Shiri Y, Bazi S, Saeidi S, Sahi Z. Antibacterial activities of Cuminum cyminum Linn. Essential Oil Against Multi-Drug resistant Escherichia coli, Int J Infect. 2014 ; 1(1):e18739. doi: 10.17795/iji-18739.

ARTICLE INFORMATION

International Journal of Infection: 1 (1); e18739
Published Online: March 31, 2014
Article Type: Research Article
Received: December 15, 2013
Revised: February 26, 2014
Accepted: March 4, 2014
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Abstract

Background: Development of antibiotic resistance among pathogenic bacteria motivates the researchers to search for newer antimicrobial agents.

Objectives: In the present study, antibacterial effects of Cuminum cyminum Linn. essential oil against multidrug resistant (MDR) Escherichia colistrains isolated from urinary tract infections were studied, using microdilution method.

Materials and Methods: A total of 12 E. colistrains were isolated from urine cultures of hospitalized patients (Zabol, southeastern Iran) suffering from urinary tract infection during 2011-2012. After bacteriological confirmatory tests, minimum inhibitory concentrations (MICs) of the essential oil of C. cyminum Linn. were determined using microdilution method. Essential oil of C. cyminum Linn. was obtained by hydro-distillation and the MICs were investigated to characterize the oil antimicrobial activities.

Results: All of E. coli isolates were resistant to four of the antibiotics including ceftazidime (50%), cefixime (41.6%), tetracycline (75%) and erythromycin (58.3%). The highest MIC value (250 ppm) was observed against two antibiotics and the lowest (10 ppm) against one antibiotic.

Conclusions: C. cyminum Linn essential oil has a potent antimicrobial activity against E. coli MDR strains. Our study confirms the use of this essential oil as an antibacterial agent. However, further research will be required before its therapeutic application.

Keywords

Escherichia coli Oils, Volatile Cuminum

Copyright © 2014, Infectious Diseases and Tropical Medicine Research Center. 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.

1. Background

The currently used feed additives such as antibiotics, probiotics and prebiotics in broiler diets to enhance nutrient utilization, play important roles in antibiotic resistance development among pathogens and saprophyte bacteria (1). The search for components with antimicrobial activities has recently gained increasing importance, due to the growing worldwide concern about the alarming increase in the rate of infections by antibiotic-resistant microorganisms. Due to their antibacterial activities against bacterial pathogens, medicinal plants are very important in human health. Cumin (Cuminum cyminum L.) originates from Egypt and Ethiopia and is much cultivated in Arabia, Malta, Sicily, India and China. Cumin seeds are used to flavor foods and liquors and its oil is utilized in perfumes and cosmetics. C. cyminum Linn. is an annual plant of the Umbelliferae family with antioxidant, anticholesterol and antimicrobial properties. Urinary tract infections (UTIs) are the second most common type of infections in the body and E. coli is the most common bacterial pathogen causing UTI (2). Different antimicrobial agents with high levels of activity against Gram-negative bacilli, including amikacin, ciprofloxacin, fosfomycin, gentamicin and nitrofurantoin, have shown acceptable levels of activities against this bacterium. Unfortunately, rapid appearance and development of drug resistance among these bacteria have caused a lot of difficulties in the modern world.

2. Objectives

In the present study, antibacterial activities of C. cyminum Linn. essential oil against multidrug resistant (MDR) E. coli strains isolated from UTIs were determined, using microdilution method.

3. Materials and Methods

3.1. Isolation of Bacteria

A total of 12 strains of E. coli were isolated from urine cultures of hospitalized patients (Zabol, southeastern Iran) suffering from UTIs, during 2011-2012. The isolated bacteria were identified and evaluated by Gram staining and standard biochemical tests (3).

3.2. Antibiotic Susceptibility Test

Susceptibility to all antibiotics was tested using standard disc diffusion method, as recommended by Clinical and Laboratory Standards Institute (CLSI) (4). Briefly, a colony suspension was prepared using sterile normal saline, equivalent to the 0.5 McFarland standard, and spread over the Mueller Hinton agar plate. Afterwards, the antibiotic discs were transferred aseptically to the surfaces of the inoculated media plates. Antibiotics and their concentrations were as follows: ceftazidim: 30 μg, tetracycline: 30 μg, erythromycin: 15 μg and ceftazidime: 30 μg. E. coli ATTCC 25922 was used as the control strain.

3.3. Plant Materials

Seeds of C. cyminum Linn. were collected from suburban areas of Kerman (southeastern Iran) and dried at room temperature in Kerman Azad University herbarium. Afterwards, the samples were crashed, transferred into a glass container, and preserved until the extraction process was performed in the laboratory.

3.4. Distillation of Essential Oil

The seeds were ground prior to the operation, and then, 300 g of the ground powder was submitted to water distillation for four hours, using a Clevenger apparatus. The distilled essential oil was dried over anhydrous sodium sulfate, filtered, and stored at 4ºC.

3.4. Minimum Inhibitory Concentration of Essential Oil

The broth microdilution method was used to determine the minimum inhibitory concentration (MIC). Briefly, serial double dilutions of the extract in Mueller Hinton broth, containing 0.5% (V/V) tween 80 over the ranges of 250, 100, 50 and 10 ppm, were prepared and added to a 96-well microtiter plate. To each well, 10 μL of indicator solution and 10 μL of Mueller Hinton broth were added. Finally, 10 μL of bacterial suspension (106 CFU/mL) was added to each well to achieve a concentration of 104 CFU/mL of the bacteria. The plates were wrapped loosely with cling film to prevent the bacteria dehydration. The plates were prepared in triplicates, and placed in an incubator at 37°C for 18–24 hours. MIC was defined as the lowest concentration of the essential oil, at which the microorganism did not demonstrate any visible growth. Average of the three values was calculated to provide the MIC values for the tested extract.

4. Results

4.1. Antibiotic Susceptibility

Antibiotic full resistance profile of the E. coli isolates was as follows: tetracycline (75%), erythromycin (58.4%), ceftazidime (50%) and cefixime (41.7%). Moderate resistance was observed in only 25% and 8.3% of the isolates against erythromycin and cefixime, respectively (Table 1). About 25% of the E. coli isolates showed resistance to all the antibiotics, whereas 8.3% and 25% showed resistance to three and two antibiotics, respectively (Table 2).

Table 1. Antimicrobial Susceptibility of Escherichiacoli isolates a
CeftazidimeErythromycinCefiximeTetracycline
Sensitive6 (50)2 (16.6)6 (50)3 (25)
Intermediate03 (25)1 (8.3)0
Resistant6 (50)7 (58.4)5 (41.7)9 (75)

a Data are presented in No. (%).

Table 2. Antimicrobial Susceptibility and Minimum Inhibitory Concentration of the Essential Oil for Escherichiacolia, b
Bacterial CodeAntibiotic SusceptibilityMIC for Essential Oil, ppmResistance Pattern
A1A2A3A4
1RRRR100A1, A2, A3, A4
2RIRS250A1, A3
3SRRRNOA2, A3, A4
4RRRR100A1, A2, A3, A4
5RRRR50A1, A2, A3, A4
6SSIS50-
7SSSS10-
8ISSR100A4
9RSSR250A1, A4
10RSIR100A1, A4
11SSSRNOA4
12RRRR100A1, A2, A3, A4

a Abbreviations: I, intermediate; MIC, minimum inhibitory concentration; NO, Any inhibition; R, resistant; S, sensitive.

b A1, erythromycin; A2, cefixime; A3, ceftazidime; A4, tetracycline.

4.2. Minimum Inhibitory Concentration Assessment for Essential Oil

Different inhibitory effects of essential oil against most E. coli isolates were demonstrated in Table 2. The essential oil had inhibitory effects against most of the isolates. About 8.3% and 16.6% of the E. coli isolates showed the lowest MICs (10 and 50 ppm, respectively), while moderate (100 ppm) and highest (250 ppm) MIC values were seen in 41.6% and 16.6% of the isolates, respectively.

5. Discussion

In the present study, E. coli strains were resistant to four of the agents, including tetracycline (75%), erythromycin (58.3%), ceftazidime (50%), cefixime (41.6%). Different results were reported by other investigators in different geographical areas. For example, Shayan et al. reported antibiotic susceptibility of the AmpC-producing E. coli isolates as follows: erythromycin (92.3%), tetracycline (92.2%) nalidixic acid (84.6%), cefixime (84.6%), difloxacin (84.6%) azithromycin (76.9%), amoxicillin (76.9%), trimethoprim-sulfamethoxazole (76.9%) and gentamicin (76.9%) (5). Madani et al. reported antimicrobial resistance to ampicillin (91.4%), cotrimoxazole (61.1%), cefixime (46.8%), gentamicin (43.3%), ceftazidime (38.8%) and nalidixic acid (38.5%) (6). In the study of Heidari-Soureshjani et al. the highest resistance was reported to ampicillin (85.71%), nalidixic acid (78.78%), and ciprofloxacin (46.51%) (7). In the recent years, essential oils of plants have been in high demand from the manufacturers of foods flavoring, fragrance, cosmetics, and pharmaceutical industries, due to the growing interest of consumers to ingredients from natural sources. In our study, 8.3% and 16.6% of the E. coli isolates showed the lowest MICs (10 and 50 ppm respectively), while moderate (100 ppm) and highest (250 ppm) MIC values were seen in 41.6% and 16.6% of the isolates, respectively. Inhibitory effects of Cumin extract on E. coli 0:157 has been demonstrated previously (8). Other authors have also shown the antimicrobial activities of hexane extract and volatile components (9), water extracts or juices (10), and methanolic extracts of C. cyminum against different bacterial strains. For example, in the study of Vaishnavi et al. Cumin seeds were effective at lower concentration against Salmonella typhi and E. coli O:157 isolates (11). Soniya et al. reported the largest diameter of inhibition zone related to methanol extracts of C. cyminum against Bacillus subtilis, E. coli and Proteus sp. (12). As the study of Steffanini et al. reported, essential oil of C. cyminum was active against different Gram-negative bacteria, including E. coli, Pseudomonas. aeruginosaandSalmonella sp. with inhibitory zones of 18 mm, 10 mm and 23 mm, respectively (13). Con et al. reported that Cumin had inhibitory effect against Staphylococcus. aureus and Micrococcus luteus (14). Akgul and Kivanc reported that Cumin exhibited an inhibitory effect against S. aureus, Klebsiella pneumonia and P. aeruginosa (15). essential oil of C. cyminum can be used for protection against some bacteria.

Acknowledgements

Footnotes

References

  • 1.

    Fuller R. Probiotics in man and animals. J Appl Bacteriol. 1989; 66(5) : 365 -78 [DOI][PubMed]

  • 2.

    Iroha IR, Adikwu MU, Esimone CO, Aibinu I, Amadi ES. Extended spectrum beta lactamase (ESBL) in E. coli isolated from a tertiary hospital in Enugu State, Nigeria. Pak J Med Sci. 2009; 25(2) : 279 -82

  • 3.

    Forbes BA, Sahm DF, Weissfeld AS. Bailey & Scott`s diagnostic microbiology. 2007; : 323 -33

  • 4.

    Reller LB, Weinstein M, Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clin Infect Dis. 2009; 49(11) : 1749 -55

  • 5.

    Shayan S, Bokaeian M, Shahraki S, Saeidi S. Prevalence of AmpC and ESBL Producing E. coli and Antibacterial Effect of Allim sativum on Clinical Isolates Collected from Zahedan Hospitals. Zahedan J Res Med Sci. 2014; 16(4) : 6 -10

  • 6.

    Madani SH, Khazaee S, Kanani M, Shahi M. Antibiotic resistance pattern of E. coli isolated from urine culture in Imam Reza Hospital Kermanshah-2006. J Kermanshah Univ Med Sci. 2008; 12(3) : 287 -95

  • 7.

    Heidari-Soureshjani E, Heidari M, Doosti A. Epidemiology of urinary tract infection and antibiotic resistance pattern of E. coli in patients referred to Imam Ali hospital in Farokhshahr, Chaharmahal va Bakhtiari, Iran. J Shahrekord Univ Med Sci. 2013; 15(2) : 9 -15

  • 8.

    Sagdic O, kuscu A, Ozcan M, Ozcelik S. Effects of Turkish spice extracts at variousconcentrations on the growth of Escherichia coli O157:H7. Food Microbiol. 2002; 19 : 473 -80 [DOI]

  • 9.

    Agnihotri S, Vaidya AD. A novel approach to study antibacterial properties of volatile components of selected Indian medicinal herbs. Indian J Exp Biol. 1996; 34(7) : 712 -5 [PubMed]

  • 10.

    Kumral A, Sahin T. Effects of some spice extracts on Escherichia coli,Salmonella typhimurium, Lister iamonocytogenes, Yersinia enterocolitica and Enterobacter aerogenes. Ann Microbiol. 2003; 53 : 427 -35

  • 11.

    Vaishnavi C, Kaur S, Kaur M. Bactericidal activity of kitchen spices and condiments on enteropathogens. Nat Prod Radiance. 2007; 6(1) : 40 -5

  • 12.

    Soniya M, Kuberan T, Anitha S, Sankareswari P. In vitro antibacterial activity of plant extracts against Gram positive and Gram negative pathogenic bacteria. Int J Microbiol Immunol. 2013; 2 : 1 -5

  • 13.

    Antimicrobial activity of the essential oils of some spice herbs. International Conference on Medicinal and Aromatic Plants (Part II) 597. : 215 -6

  • 14.

    Con AH, Ayar A, Gokalp HY. Antimicrobial activity of the essentialoils extracted from some spices. Food. 1998; 23 : 171 -5

  • 15.

    AkguL A, Kivanc M. Growth of Staphylococcus Aureus in Köfte, a Turkish Ground Meat Product, Containing Laser Trilobum Spice. J Food Safety. 1989; 10(1) : 11 -9 [DOI]

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