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 Table of Contents  
ORIGINAL ARTICLE
Year : 2015  |  Volume : 6  |  Issue : 1  |  Page : 44-46

Antibacterial potency of Ziziphus mauritiana (Fam-Rhamnaceae) roots


1 Department of Pharmacognosy, School of Pharmaceutical Sciences, IFTM University, Moradabad, India
2 BBS Institute of Pharmaceutical and Allied Sciences, Greater Noida, Uttar Pradesh, India

Date of Web Publication8-Jan-2015

Correspondence Address:
Sukirti Upadhyay
School of Pharmaceutical Sciences, IFTM University, Moradabad, Uttar Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2394-2002.148895

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  Abstract 

Background: Ziziphus mauritiana is a well known herb generally its fruit is used in stomach ailments or infections.The genus Ziziphus is known for its antimicrobial potency. Material and Methods: Here Z. mauritiana roots were taken for conducting experiments in various bacterial strains for evaluating its antibacterial potency by disc diffusion method. The minimum inhibitory concentration and zone of inhibition were also determined. The mode of action was determined by leakage of nucleic acid and its determination by UV spectrometry. Results: The root extract of Z.mauritiana posses excellent antibacterial activity against as the activity is well compared with standard drug ciprofloxacin. The mode of action of the extract was bactericidal. Conclusion: Effective and safe herbal antibiotic may be produced in the future by root extract of Z.mauritiana.

Keywords: Disc diffusion, minimum inhibitory concentration, Ziziphus mauritiana, zone of inhibition


How to cite this article:
Upadhyay S, Mishra A, Srivastava S, Upadhyay P, Ghosh A K, Singh V. Antibacterial potency of Ziziphus mauritiana (Fam-Rhamnaceae) roots. Drug Dev Ther 2015;6:44-6

How to cite this URL:
Upadhyay S, Mishra A, Srivastava S, Upadhyay P, Ghosh A K, Singh V. Antibacterial potency of Ziziphus mauritiana (Fam-Rhamnaceae) roots. Drug Dev Ther [serial online] 2015 [cited 2017 Aug 24];6:44-6. Available from: http://www.ddtjournal.org/text.asp?2015/6/1/44/148895


  Introduction Top


The wide use of antibiotics in the treatment of bacterial infections has led to the emergence and spread of resistant strains. Emergence of multiple drug resistance to human pathogenic organisms has necessitated a search for new antimicrobial substance from other sources including. The genus Ziziphus belongs to the family Rhamnaceae. This genus comprises of about 100 species of deciduous or evergreen trees and shrubs distributed in the tropical and subtropical regions of the world. Ziziphus species can grow either as shrublets, shrubs or trees with thorny branches and are used as a hedge to form defensive fences for animals. [1] Ziziphus mauritiana Lam. occurs on nearly every continent. Z. mauritiana have very nutritious fruits and are usually eaten fresh. [2] Root extract of Z. mauritiana possess antidiarrhoeal activity in rodents in castor oil induced diarrhoea. [3] Aqueous extract of Z. maurtiana leaf can be used for the prevention and treatment of fatty liver, atherosclerosis and other diseases associated with high levels of cholesterol and triglyceride and pretreatment was found to confer more protection than co-treatment, hence pretreatment should be preferred. [4] The methanolic extract of Z. mauritiana stem bark possess antiulcer activity. [5] Investigation of the MeOH extract revealed that isolated alkaloids exhibited potent antiplasmodial activity against the parasite Plasmodium falciparum. [6] Alkaloidal fraction of the drug also showed positive effect in increasing sexuality and fertility of male wistar albino rats. [7] Recently antimicrobial potency of Z. mauritiana leaves is explored. [8] Root generally contain antimicrobial agent so present study was undertaken to determine antibacterial activity of Z. maurtiana roots.


  Materials and Methods Top


Plant material

Dried roots were collected from deer park, Moradabad in February 2010 and identified by Dr. DV Amla, of National Botanical Research Institute (NBRI), Lucknow, Uttar Pradesh, India, and a voucher specimen (no. NBRI-SOP - 202) was preserved in NBRI for future references.

Solvent extraction

Ziziphus mauritiana was extracted with dichloromethane (DCM) in Soxhlet extractor for 48 h. The solvent extracts were concentrated under reduced pressure and the yield was 10%.

Phytochemical analysis of plant extracts for active components

The plant extract was analysed for phytoconstituents group using standard procedures. [9]

Bacterial strains

Gram-positive bacteria, Bacillus cereus (National collection of industrial microbiology [NCIM]-2016), Staphylococus aureus (NCIM-2079) and Gram-negative bacteria, Escherichia coli (NCIM-2089) and Pseudomonas aeruginosa (NCIM 2200) strains NCIM, National Chemical Laboratory, Pune, India.

Maintenance of test microorganism for antimicrobial studies

The bacteria were maintained on nutrient broth (NB) at 37°C.

Preparation of inoculum

The Gram-positive (Bacillus subtilis and S. aureus) and Gram-negative bacteria (E. coli, P. aerigonasae were precultured in NB.

Antibacterial activity

The root extract of Z. mauritiana, was tested by the disc diffusion method. Different concentration of the extracts was prepared by reconstituting with dimethylsulphoxide (DMSO).

Disc diffusion method

The in vitro antibacterial activity of the DCM extract of Z. mauritiana roots was carried out by disc diffusion method. [10] Actively growing cultures were inoculated in agar - nutrient media in different petridishes. The whatman paper disc of impregnated with various extract concentrations of extract, DMSO alone (control), and an antibiotic ciprofloxacin (standard). Then they were incubated for 24 h at 37°C. [11]

Minimum inhibitory concentration

The minimum inhibitory concentration (MIC). [12] was determined using tube dilution technique. Varying concentrations of the extracts were prepared and 1 ml introduced into 9 ml of NB in test tubes. About 0.1 ml of the culture diluted to 10 5 CFU/ml was added and incubated accordingly. The least concentration of the extract that did not permit turbidity in the broth was taken as the MIC.

Mode of action

The method of Heipieper et al. [13] was followed to determine the leakage of ultraviolet (UV) 260 and UV 280 absorbing material. The bacterial suspension was prepared. P. aeruginosae cells were grown overnight with continuous shaking in NB at 37°C, harvested, washed with 10 mm ethylenediaminetetraacetic acid and then twice in distilled water by centrifugation each time 6000 rpm for 15 min at 4°C and resuspended such that absorbance of the final suspension was 2.0 at A450. After incubation for 30 min at room temperature, the test compound was added to suspension at MIC. At regular interval of 15 min regular aliquots of the sample were drawn centrifuged and UV 260 and UV 280 absorbing material in the suspension was measured.


  Results Top


Phytochemical analysis shows that extract contain alkaloids, phenolics, glycosides saponin and steroids.

The antibacterial activity of the extract was performed using four bacterial strains by disc diffusion method as compared to standard drug ciprofloxacin extract possess excellent inhibitory activity against both Gram-positive and negative bacteria in 0.1 μL [Table 1] concentration although it is somewhat less than standard drug ciprofloxacin but being natural alternative it is more preferable.
Table 1: The antibacterial activity of the extract


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Minimum inhibitory concentration

The extract shows minimum inhibitory concentration at value of 1 ppm for Gram-negative bacteria and 2 ppm for Gram-positive bacteria [Table 2].
Table 2: Bacterial strain and MIC


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Mode of action

Ultraviolet spectophotometry showed increase in concentration of nucleic material as increase in absorbance at 260 nm with time. This happens due to increase in nucleic material in the aliquots with time due to bactericidal effect of extract by leakage of nucleic material of bacteria [Table 3].
Table 3: Time versus leakage of nucleic acid


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  Discussion Top


Extract of Z. mauritiana roots shows excellent activity against both Gram-positive (B. subtilis and S. aureus) and Gram-negative bacteria (E. coli, P. aerigonasae) and the extract exhibit its effect in dose dependent manner genus Ziziphus possess antimicrobial property. Here in this extract presence of phenolic content also indicate its antimicrobial potency. It shows half MIC for Gram-negative bacteria than Gram-positive bacteria. Firstly, if the floral extract disrupts lipopolysaccharide of bacterial strain, it may help restoring a porins channel thus facilitating the flow of ciprofloxacin to target sites. Secondly, it may cause negative effects on the efflux mechanism and let the sufficient concentration of ciprofloxacin to remain in bacterium thus helping in its increased antimicrobial.

Activity [14],[15],[16] thirdly, Cryptocoryne auriculata flower extract may be inhibiting protein synthesis as while acting alone it inhibits the growth of bacterial strains tested. It shows increase leakage of nucleic acid from cell death from increasing time.


  Conclusion Top


This study concluded that root extract of Z. mauritiana possess antibacterial activity may be due to phenolic content although somewhat less then synthetic standard drug so formulations may be prepared with this extract as alternative to synthetic drugs especially for children and weak persons.

 
  References Top

1.
www. ayurvedicmedicinalplants. Com.  Back to cited text no. 1
    
2.
Upadhyay S, Upadhyay P, Ghosh AK, Singh V. Ziziphus mauritiana : A review on pharmacological potential of this underutilized plant. International Journal of Current Research and Review 2012; 04:141-44.  Back to cited text no. 2
    
3.
Dahiru D, Sini JM. and John A L. Antidiarrhoeal activity of Ziziphus maurtiana root extract in rodents. African Journal of Biotechnology 2006; 5:941-945.  Back to cited text no. 3
    
4.
Dahiru D, Obidoa O, Effect of aqueous extract of Ziziphus maurtiana leaf on cholesterol and triglycerides level in serum & liver of rats administered alcohol. 2009, Pakistan j of nutrition; 2009; 1884-1888.  Back to cited text no. 4
    
5.
Panchal S, Panchal K, Vyas N, Modi K, Patel V, Bharadia P, Pundarikakshudu K. Antiulcer Activity of Methanolic Extract of Z. mauritiana stem Bark. International Journal of Pharmacognosy and Phytochemical Research; 2010; 2:6-9:1884-1888.  Back to cited text no. 5
    
6.
Panomwan P, Kanlaya L., Samran P, Palangpon K et al. Antiplasmodial and antimycobacterial cyclopeptide alkaloids from the root of Ziziphus mauritiana, Phytochemistry 2011; 72:909-915  Back to cited text no. 6
    
7.
Upadhyay S, Upadhyay P, Ghosh AK, Singh V. Effect of ethanol extract and alkaloidal fraction of Ziziphus mauritiana (Fam-Rhamnaceae) roots on fertility and sexual behavior of male wistar albino rats 2013;2:12-18.  Back to cited text no. 7
    
8.
Abalaka M E, Daniyan1 S Y, Mann A Evaluation of the antimicrobial activities of two Ziziphus species (Ziziphus mauritiana L. and Ziziphus spinachristi L.) on some microbial pathogens African Journal of Pharmacy and Pharmacology April 2010; 4:135-139.  Back to cited text no. 8
    
9.
Kokate CK. Practical Pharmacognosy, 4 th edition, Vallabh Prakashan, New Delhi, 1994; p 4-29.  Back to cited text no. 9
    
10.
Elgayyar M, Draughon FA, Golden DA, Mount JR. Antimicrobial activity of essential oils from plants against selected pathogenic and saprophytic microorganisms. J Food Prot 2001; 64:1019-24.  Back to cited text no. 10
    
11.
Lakshmi B O, Karsha P V. Antibacterial activity of Black pepper(Piper nigrum Linn) with special reference to its mode of action on bacteria. Indian j of nat prod and resourses, June 2010; 1:213-215.  Back to cited text no. 11
    
12.
Stokes EJ, Clinical bacteriology, Edward Arnold ltd, London; 1975; p 208.  Back to cited text no. 12
    
13.
Heipieper HJ, Diefenbach R, Keweloh H. Conversion of cis unsaturated fatty acids to trans, a possible mechanism for the protection of phenol-degrading Pseudomonas putida P8 from substrate toxicity. Appl Environ Microbiol 1992;58:1847-52.  Back to cited text no. 13
    
14.
Stermitz FR, Lorenz P, Tawara JN, Zenewicz LA, Lewis K. Synergy in a medicinal plant: Antimicrobial action of berberine potentiated by 5'-methoxyhydnocarpin, a multidrug pump inhibitor. Proc Natl Acad Sci U S A 2000; 97:1433-7.  Back to cited text no. 14
    
15.
Gibbons S, Udo EE. The effect of reserpine, a modulator of multidrug efflux pumps, on the in vitro activity of tetracycline against clinical isolates of methicillin resistant Staphylococcus aureus (MRSA) possessing the tet(K) determinant. Phytother Res 2000; 14:139-40.  Back to cited text no. 15
    
16.
Van Bambeke F, Balzi E, Tulkens PM. Antibiotic efflux pumps. Biochem Pharmacol 2000; 60:457-70.  Back to cited text no. 16
    



 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

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Abstract
Introduction
Materials and Me...
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