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ORIGINAL ARTICLE
Year : 2020  |  Volume : 9  |  Issue : 3  |  Page : 229-232

Antimicrobial activity of Cinnamomum zeylanicum aqueous extract against bacteria and fungi responsible for urinary tract infection


1 Department of Clinical Nutrition, SDNB Vaishnav College, Chennai, Tamil Nadu, India
2 Department of Home Science, Women's Christian College, Chennai, Tamil Nadu, India

Date of Submission26-Jan-2020
Date of Decision30-Apr-2020
Date of Acceptance20-May-2020
Date of Web Publication28-Jul-2020

Correspondence Address:
Dr. Sivapriya Thiyagarajan
Department of Clinical Nutrition, SDNB Vaishnav College, Chrompet, Chennai- 600 044, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijhas.IJHAS_3_20

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  Abstract 


BACKGROUND: In this modern age of multidrug-resistant pathogens, Cinnamomum zeylanicum has a promising antimicrobial activity against urinary pathogens. The study was conducted to determine the antimicrobial activity of cinnamon against the bacterial family Enterobacteriaceae strains and Fungi strains and compare it with standard antibiotics.
MATERIALS AND METHODS: Standard agar well technique was executed to obtain the zone of inhibition (ZOI). Mueller–Hinton agar was prepared and left to cool at 45°C. The agar medium was poured into the Petri plates, and the medium was preseeded with cultures using a sterile cotton swab dipped in the inoculum. Wells of 6 mm were dug and the dug wells were then filled with 50, 100, and 150 μl of aqueous extract of Cinnamon zeylanicum and incubated at 37°C for 24 h. ZOI was determined and compared with standard antibiotics.
RESULTS: Urinary pathogen, Staphylococcus aureus (ZOI-24 mm), was extremely sensitive to C. zeylanicum aqueous extract at 150 μl concentration, followed by Salmonella typhi (ZOI-22 mm) and Escherichia coli (ZOI-20 mm). Enterobacter spp. was also sensitive and demonstrated ZOI of 14 mm at 150 μl concentration.
CONCLUSION: The ZOI for pathogens with various concentrations of C. zeylanicum was comparatively significant and demonstrated its potential use as an antimicrobial agent with an efficacy that can be compared to that of the already recognized and widely used antibiotics, namely, cephalexin and amikacin.

Keywords: Antibiotics, bacteria, Cinnamon, fungi, urinary tract infection, zone of inhibition


How to cite this article:
Thiyagarajan S, John S. Antimicrobial activity of Cinnamomum zeylanicum aqueous extract against bacteria and fungi responsible for urinary tract infection. Int J Health Allied Sci 2020;9:229-32

How to cite this URL:
Thiyagarajan S, John S. Antimicrobial activity of Cinnamomum zeylanicum aqueous extract against bacteria and fungi responsible for urinary tract infection. Int J Health Allied Sci [serial online] 2020 [cited 2020 Aug 6];9:229-32. Available from: http://www.ijhas.in/text.asp?2020/9/3/229/290717




  Introduction Top


Extensive use of antibiotics in clinical medicine promotes the development of antibiotic resistance among infectious microbial strains. This development results in a very serious problem in the treatment of pathogenic microbes, which has led to the search of new antimicrobial agents.[1]

Drug-resistant pathogens are one of the key public health challenges of the 21st century. The most critical group of multidrug-resistant bacteria that pose a particular threat in hospitals, nursing homes, and among patients are various Enterobacteriaceae, including Klebsiella,  Escherichia More Details coli, Serratia, and Proteus. These bacteria have become resistant to a large number of antibiotics, including carbapenems and third-generation cephalosporins.[2]

Many hundreds of plants worldwide are used in traditional medicine as treatments for bacterial infections. Conventional drugs usually provide effective antibiotic therapy for bacterial infections, but there is an increasing problem of antibiotic resistance and a continuing need for new solutions. Although natural products are safer than synthetic antibiotics, health-care professionals should be aware of the available evidence for herbal antibiotics.[3]

Urinary tract infection is the most common infection among diabetes patients. The bacterial family Enterobacteriaceae are the most frequent pathogens detected, causing 84.3% of urinary tract infection. Among the pathogens causing urinary tract infection, E. coli are the predominant pathogens (80%), followed by Staphylococcus (10%–15%). The pathogens traditionally associated with urinary tract infection change many of their features because of antimicrobial resistance.[4]

Cinnamon has the most potential bactericidal properties.[5] Cinnamaldehyde in cinnamon has been shown to destroy the cytoplasmic membrane of both Gram-positive and Gram-negative bacteria and induce depletion of the intracellular adenosine triphosphate (ATP) concentration.[6] Furthermore, cinnamaldehyde inhibits amino acid decarboxylation activity within the cells. This inhibitory action leads to energy deprivation within the microbial cell and death.[7]

The antifungal effects of over 1300 species have been studied and it was concluded that cinnamon exhibited the highest antifungal property. The antimicrobial substances contained in the extract including alkaloids, terpenoids, and flavonoids serve as a source of drugs against urinary pathogens. Providing its low toxicity, utilization of this compound may be expected as an antifungal agent.[8]

Currently available synthetic antifungal drugs act by inhibition of macromolecule synthesis of fungi (flucytosine), impairment of membrane barrier function (polyenes), inhibition of ergosterol biosynthesis (allylamines, thiocarbamates, azole derivatives, and morpholines), interaction with microtubules (griseofulvin), and inhibition of b-1,3-glucan synthesis (echinocandins). Unfortunately, they have major side effects on host tissues; amphotericin B is used to treat disseminated candidiasis, but it showed nephrotoxicity, reduction in renal blood flow, nausea, vomiting, and anorexia.[9] Hence, to combat the effects of drug-resistant pathogens, the objective of the present study was framed to determine and compare the antimicrobial activity of Cinnamomum zeylanicum, against urinary pathogens.


  Materials and Methods Top


Agar well diffusion method for Enterobacteriaceae

The study was conducted in the biotechnology of Women's Christian College and the study was commenced after getting ethics approval from the independent institutional ethics committee. Dried aqueous extract powder of C. zeylanicum was purchased with certificate of analysis from Nava Chetna Kendra, New Delhi, and the aqueous extract was prepared by boiling 1 g of powder in 10 ml of water and filtering it. The extract 0.1 g/ml was used for determining antimicrobial activity.

As most number of bacteria isolated from urine of patients with urinary tract infection belong to the family of Enterobacteriaceae, the family Enterobacteriaceae was selected for the study. The bacterial strains investigated were from the family Enterobacteriaceae that included E. coli, Klebsiella pneumoniae, Staphylococcus, Enterobacter spp., Pseudomonas aeruginosa,  Salmonella More Details typhi, and Shigella flexneri. The cultures were grown on Tryptone soya broth at 37°C for 24 h in test tubes in an incubator. The turbidity was measured by adjusting to 0.5 McFarland standards (108 CFU/ml).

Standard agar well technique was executed to obtain the zone of inhibition (ZOI). Mueller–Hinton agar was prepared and left to cool at 45°C. The agar medium was then poured into the Petri plates, and the medium was preseeded with different cultures using a sterile cotton swab dipped in the inoculum. The well puncture was dipped in ethanol and sterilized by flaming. Wells of 6 mm were dug and the dug wells were then filled with 50, 100, and 150 μl of aqueous extract of C. zeylanicum (AECZ). The plates were incubated at 37°C for 24 h. Antibiogram was performed using antibiotics, namely cephalexin and amikacin, to compare the ZOI with those of sample extract.[10]

Disk diffusion method for antifungal assay

The antifungal activity of AECZ was tested on Aspergillus niger and Candida albicans by disk diffusion technique and compared with standard ketoconazole. The ZOI formed for the AECZ and ketoconazole against pathogens was calculated. The method followed by Ahmad et al., 2013, was used to determine the antifungal activity. Potato dextrose agar (PDA) was used for fungal cultures. The culture medium was inoculated with the fungal strains separately suspended in potato dextrose broth and this suspension was poured on 15 ml solidified PDA plates. The plates were allowed to cool and solidify. The wells of diameter 6 mm were punched into the agar. The test extracts and the antibiotic ketoconazole 30 μg were added to these wells. The plates were kept for incubation at 37°C for 48–72 h. The fungal growth was observed and the results were interpreted.


  Results Top


Test pathogens in the present investigation included E. coli, K. pneumoniae, P. aeruginosa, Staphylococcus aureus, S. typhi, S. flexneri, and Enterobacter spp. These pathogens cause urinary tract infection among diabetes individuals. The results were observed after incubating the Petri plates at 37°C for 24 h.

ZOI is the area where growth of the microorganism is inhibited by the antimicrobial agent within a lawn of microorganism on a solid medium. A larger ZOI around the antimicrobial agent indicates that the microorganisms are more sensitive to the antibiotic. The sensitivity of the cinnamon extract, classified by the diameter of the inhibition zone, as per the procedure of Castro and Lima and Srivastava,[11],[12] is presented in [Table 1].
Table 1: Comparison of the zone of inhibition exhibited by AECZ and antibiotics against urinary pathogens

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Comparison of ZOI between cinnamon, cephalexin and amikacin by agar well diffusion method.

Zone of inhibition exhibited by aqueous extract of Cinnamomum zeylanicum

Urinary pathogen, S. aureus (ZOI-24 mm), was extremely sensitive to C. zeylanicum aqueous extract at 150 μl concentration, followed by S. typhi (ZOI-22 mm) and E. coli (ZOI-20 mm). Enterobacter spp. was sensitive and demonstrated ZOI of 14 mm at 150 μl concentration.

Zone of inhibition exhibited by antibiotic cephalexin

Urinary pathogens, E. coli (ZOI-24 mm), S. typhi (ZOI-24 mm), and Enterobacter spp. (ZOI-24 mm), were extremely sensitive to C. zeylanicum aqueous extract at 150 μl concentration. P. aeruginosa was sensitive and exhibited ZOI of 13 mm.

Zone of inhibition exhibited by antibiotic amikacin

Enterobacter spp. (ZOI-29.5 mm), followed by Klebsiella pneumonia (ZOI-29 mm), were extremely sensitive to antibiotic amikacin at 150 μl concentration; S. flexneri was sensitive and exhibited ZOI of 12 mm.

The ZOI for pathogens with various concentrations of C. zeylanicum was comparatively significant and demonstrated the potential use of it as an antimicrobial agent with an efficacy that can be compared to that of the already recognized and widely used antibiotics, namely, cephalexin and amikacin.

Antifungal activity of aqueous extract of Cinnamomum zeylanicum

The opportunistic yeast pathogens, C. albicans and Aspergillus spp., cause life-threatening infectious diseases in immunocompromised patients, leading to a raise in the mortality rate. The existing antifungal drugs demonstrate high toxicity to host tissues causing undesirable effects. Many metabolites of terrestrial and aquatic plants contain a rich source of unexplored novel leads which can be used to treat various diseases. Such natural products are less expensive and have lower toxicity to host tissues.[13]

From [Table 2], it can be interpreted that aqueous extract of C. zeylanicum mildly inhibited the growth of pathogenic fungi. ZOI was found to be 10 mm for C. albicans and 9 mm for A. niger in 200 μl of AECZ. The antibiotic, ketoconazole, inhibited the pathogens even at smaller doses of 50 μl and the ZOI was found to be 25 mm.
Table 2: Comparison of zone of inhibition of AECZ and Ketoconazole against fungi

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C. albicans are the most pathogenic candida species and responsible for the significant increase in the number of infections. Candida strains were sensitive to the essential oil of C. zeylanicum with minimum inhibition values ranging between 312.5 and 625 μg/ml. Cinnamaldehyde in fat-soluble extract of C. zeylanicum was effective against all the five selected species of Aspergillus.[14]


  Discussion Top


The results of this study reveal that cinnamon can be used as an alternative medicine in the treatment of urinary tract infection. Previous studies consider that the antibacterial effect of cinnamon is probably due to its major compound, cinnamaldehyde. Cinnamon has the most potent bactericidal action. Cinnamaldehyde damages the cytoplasmic membrane of both Gram-positive and Gram-negative bacteria and induce the depletion of intracellular ATP concentration.[15] Extracts of spices have aroused the interest to be used as an alternative remedy for the treatment of many infectious diseases. C. zeylanicum extracts have been shown to possess antibacterial, antifungal, antiviral, insecticidal, germicidal, and antioxidant property.[16]

It has been proposed that cinnamaldehyde and eugenol inhibit production of an essential enzyme by the bacteria and/or cause damage to the cell wall of the bacteria.[17] Therefore, the high antimicrobial activity of cinnamon oil is due to the presence of the high amount of cinnamaldehyde.[18] Cinnamaldehyde may be a potential lead compound for the development of antifungal drugs through the control β-glucan and chitin synthesis in yeasts and molds.[15]


  Conclusion Top


Cinnamon has an enormous potential as an antimicrobial agent and is a powerful antioxidant. This study showed that cinnamon is a more potent antimicrobial agent than any extract and it has the potential for further research in drug development and as a food preservative when compared with standard antibiotics; cinnamon extract revealed commendable results and can be consumed as a natural antibiotic.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

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Dugoua J, Seely D, Perri D, Cooley K, Forelli T, Mills E, et al. From type 2 diabetes to antioxidant activity: A systemic review on cinnamon bark. Can J Physiolpharmacol 2007;85:837-47.  Back to cited text no. 5
    
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Rajeshkumar R, Sundararaman M. Emergence of Candida spp. and exploration of natural bioactive molecules for anticandidal therapy – Status quo. Mycoses 2012;55:e60-73.  Back to cited text no. 9
    
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Ponce AG, Fritz R, Delvalle CR. Antimicrobial activity of essential oils on the native microflora of organic Swiss chard. J Agrl Food Chem 2003;36:679-84.  Back to cited text no. 10
    
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Castro RD, Lima EO. Anti-Candida activity and chemical composition of Cinnamomum zeylanicum blume essential oil Braz. Arch Biol Technol 2013;56:749-55.  Back to cited text no. 13
    
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Srivastava N. In vitro assesment of fungitoxicity of Cinnamon bark oil against Aspergillus spp. International Journal of Pharma and Bio Sciences 2015;6:B656-63·  Back to cited text no. 14
    
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Nuryastuti T, Mei HC, Busscher HJ, Iravati S, Aman AT, Korn BP. Effect of cinnamon oil on IcaA expression and biofilm formation by Staphylococcus epidermidis. Appl Environ Microbiol 2009;75:6850-5.  Back to cited text no. 15
    
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Seenivasan S, Jayakumar M, Ignacimuthu S. In vitro antibacterial activity of some plant essential oils. BMC Complement Altern Med 2006;6:39.  Back to cited text no. 16
    
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Boyko EJ, Fihn SD, Scholes D, Abraham L, Monsey B. Risk of urinary tract infection and asymptomatic bacteriuria among diabetic and nondiabetic postmenopausal women. Am J Epidemiol 2005;161:557-64.  Back to cited text no. 17
    
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Inns T, Millership S, Teare L, Rice W, Reacher M. Service evaluation of selected risk factors for extended spectrum beta lactamase Eschericia coli urinary tract infections: A case control study. J Hosp Infect 2014;47:197-203.  Back to cited text no. 18
    



 
 
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