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ORIGINAL RESEARCH Table of Contents   
Year : 2009  |  Volume : 20  |  Issue : 3  |  Page : 337-339
Screening of Quercus infectoria gall extracts as anti-bacterial agents against dental pathogens


Department of Botany and Microbiology, Gurukul Kangri University, Haridwar, India

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Date of Submission17-Jun-2008
Date of Decision01-Dec-2008
Date of Acceptance23-Apr-2009
Date of Web Publication30-Oct-2009
 

   Abstract 

Background and Objectives: A number of bacteria have now become antibiotic-resistant. This increases the importance of ayurvedic drugs. We report, here, the activity of different extracts (petroleum ether, chloroform, methanol and water) of Quercus infectoria galls against dental pathogens - Streptococcus mutans, Streptococcus salivarius, Staphylococcus aureus, Lactobacillus acidophilus (designated) and Streptococcus sanguis (isolated).
Materials and Methods: The cup-plate method was used in anti-bacterial activity of the extracts at concentration of 200 mg/ml against dental pathogens. Minimum inhibitory concentration (MIC) values of most effective extracts against the most susceptible bacteria were determined using a two-fold serial micro dilution method.
Results: Methanolic extract showed maximum anti-bacterial activity against all the bacteria. The most susceptible bacteria were S. sanguis followed by S. aureus, S. mutans, S. salivarius and L. acidophilus. The MIC values showed that methanolic extract was more effective than water extract.
Conclusion: The plant has the potential to generate herbal metabolites. The crude extracts demonstrating anti-dental caries activity could result in the discovery of new chemical classes of antibiotics. These chemical classes of antibiotics could serve as selective agents for the maintenance of human health and provide bio-chemical tools for the study of infectious diseases.

Keywords: Anti-bacterial activity, cup-plate method, minimum inhibitory concentration, Quercus infectoria

How to cite this article:
Vermani A, Navneet, Prabhat. Screening of Quercus infectoria gall extracts as anti-bacterial agents against dental pathogens. Indian J Dent Res 2009;20:337-9

How to cite this URL:
Vermani A, Navneet, Prabhat. Screening of Quercus infectoria gall extracts as anti-bacterial agents against dental pathogens. Indian J Dent Res [serial online] 2009 [cited 2014 Sep 19];20:337-9. Available from: http://www.ijdr.in/text.asp?2009/20/3/337/57380
Quercus infectoria (Fabaceae) is a small tree. The galls arise on young branches of this tree as a result of attack by the gall-wasp, Adleria gallae - tinctoria. [1] The plant is known as Mayaphal and Majufal in Hindi. Streptococcus mutans, Streptococcus sanguis,  Streptococcus salivarius Scientific Name Search  Lactobacillus acidophilus Scientific Name Search y a major role in dental plaque formation. Staphylococcus spp. also ferments many sugars and the produced product is utilized by the dental plaque bacteria. [2] A large number of Streptococcus spp. and Lactobacillus spp. are involved in root caries and periodontal diseases. [3],[4] In recent times micro- organisms developed resistance to many antibiotics due to the indiscriminate use of anti-microbial drugs in the treatment of infectious diseases. Therefore, developing of alternative antimicrobial agents is required, and local medicinal plants are considered the important sources of novel anti- microbial agents. [5] This study was carried out to investigate the anti-bacterial properties of the galls of Q. infectoria, extracted by four solvents of different polarity against dental pathogens.


   Materials and Methods Top


The galls of Q. infectoria used in this study were obtained from the local market. They were identified at the Department of Botany and Microbiology, Gurukul Kangri University, Haridwar. The galls were crushed to small pieces using pestle and mortar and then powdered in an electric grinder. The extracts were prepared by immersing 200 gm of dried powdered material in 600 ml of solvents i.e. petroleum ether, chloroform, methanol and water using the Soxhlet apparatus. Crude extracts were obtained by removing the solvent in vacuum evaporator at 30 C and stored in sterile bottles at 4 C until further use. The strains of dental infection-related bacteria used in this study were Staphylococcus aureus Scientific Name Search  MTCC-1144, S. mutans MTCC-890, S. salivarius MTCC-1938, L. acidophilus MTCC-447 and local clinical isolate of S. sanguis (Aggarwal Dental Clinic, Haridwar). The isolates were identified according to published guidelines. [6] All the bacterial strains were grown and maintained on nutrient agar slants at 4 C. The extracts were dissolved in the same solvent with which it has been extracted (petroleum ether, chloroform, methanol and water) to a final concentration of 200 mg/ml for cup-plate method. [7]

The cup-plate method was used to evaluate the anti-bacterial activity. This method depends upon the diffusion of the tested material to such an extent that growth of the added micro-organism is prevented entirely in a zone around the hole containing a solution of tested material. [8],[9] One hundred microliters of diluted inoculum of 10 5 CFU/ml [10] of 24 hours old cultures of test organisms were mixed in Mueller Hinton Agar media and shaken. Then media was poured (25-30 ml) in sterilized petridishes (20 Χ 90 mm). Wells of 8 mm diameter were punched into the agar medium and filled with 45 μl of plant extracts. All the solvents served as negative control. Each extract was assayed in triplicate and the mean values were observed. The plates were incubated at 37 C for 24 hours. The anti-bacterial activity was interpreted from the size of the diameter of zone of inhibition measured in millimeters (mm), it was observed as the clear zones surrounding the hole.

The MIC of the most effective extracts (methanolic and aqueous) was determined for S. aureus and S. sanguis. MIC was determined by using the two-fold serial micro dilution method at a final concentration ranging from 10 mg/ml to 0.0195 mg/ ml. [11] The extracts were added to sterile Mueller Hinton Broth before bacterial suspensions with final inoculum of 10 5 CFU/ml. Each extract was assayed in triplicate. The extracts in broth were used as negative control and the bacterial suspensions were used as positive control. The turbidity of the wells in the microtiter plate was interpreted as visible growth of the microorganisms. The MIC values were taken as the lowest concentration of the extracts which showed no turbidity after 24 hours of inoculation at 37 C.


   Results Top


[Table 1] represents anti-bacterial activities of all the extracts with respect to each of the test organism at concentration of 200 mg/ml. Anti-bacterial activity was found in all the extracts. The methanolic extract showed the maximum zone of inhibition against all the bacteria. The anti-bacterial activities of extracts were observed in increasing order- petroleum ether is less than chloroform which is less than water which is further less than methanol. The methanolic extract showed the maximum zone of inhibition against S. sanguis. The minimum zone of inhibition was showed by petroleum ether extract against L. acidophilus.

The MIC values of the methanolic and aqueous extracts from the galls of Q. infectoria against S. aureus and S. sanguis are shown in [Table 2]. The MIC values of the methanolic and aqueous extracts were the same (0.1563 mg/ml) against S. aureus, whereas the MIC values of methanolic and aqueous extracts against S. sanguis were 0.0781 mg/ml and 0.1563 mg/ml.


   Discussion Top


Streptococci and staphylococci ferment sugars and produce acids. These acids affect primary decalcification of enamel which leads to total destruction and the decalcification of dentin. Major end-products of fermentation are lactic acid, dextrans and levans. These products are seen to cause dental caries. [2] Present study has shown that the gall is potentially a rich source of anti-bacterial agents. All the four extracts inhibited the growth of all pathogens and methanolic extract was the most effective. The MIC value of methanol and water extracts against S. sanguis in comparison to S. aureus concludes that S. sanguis showed greater sensitivity towards the methanolic extract.

Successful prediction of botanical compounds from plant material is largely dependent on the type of solvent used in the extraction procedure. Traditional healers use primarily water as the solvent. [8] However, in the present study, we have found that plant extract in methanol provided more consistent antimicrobial activity as compared to those extracted in water. It is probably because various organic compounds can be leached more in this solvent. Ethanolic extract of Q. infectoria also demonstrated significant activity against resistant bacteria. [12],[13] Ahmad et al. [8] screened medicinal plants to detect anti-microbial activity and clearly demonstrated that alcohol is a better solvent as compared to water and petroleum ether.

Prabhat et al. [9],[14] reported good anti-bacterial activity of Achyranthes aspera (Apamarga) and Mimusops elenigi (Bakula) methanolic extracts against these dental pathogens. However, in this study, the gall extract showed better results than both the plants. The main constituents found in the galls of Q. infectoria are tannin (50-70%), gallic acid and ellagic acid. [11],[15] Tannin is a phenolic compound that is soluble in water, alcohol and acetone. It gives precipitates with protein. [16] The anti-microbial activity seemed to depend on the contents of tannin and gallic acid in the extracts. [17],[18]

In conclusion, extracts of galls of Q. infectoria have high potential as anti-bacterial agent. This finding provides an insight into the usage of the galls of Q. infectoria as traditional medicine in dental powder and in the treatment of toothache and gingivitis. [19],[20] The results of this study suggest that plant extracts possess compounds with anti-bacterial properties which can be used as anti-microbial agents in new drugs for the therapy of infectious diseases.

Further pharmacological evaluation of refined extracts of galls of Q. infectoria are needed before they can be used as therapeutic antimicrobials.

Further, phyto-chemical studies are required to establish the types of compounds responsible for the anti-microbial effects of this medicinal plant.


   Acknowledgment Top


I am also thankful to Botanical Survey of India, Dehradun, for help during identification of plant.

 
   References Top

1.Samuelsson RG. Drugs of natural origin. A textbook of Pharmacognosy. Apote Karsociatetion, 4 th ed. Sweden: Swedish Pharmaceutical Press; 1999.  Back to cited text no. 1      
2.Kornman KS, Robertson WJ. Clinical and microbiological evaluation of therapy for Juvenila periodontitis. J Periodontol 1985;56 : 443-56.  Back to cited text no. 2      
3.Schiipbach P, Osterkwalder V, Guggenheim B. Human root caries microbiota in plaque covering sounds carious and arrested carious root surfaces. Caries Res 1995;29 : 382-95.  Back to cited text no. 3      
4.Marsh PD. Microbiological aspects of the chemical control of plaque and gingivitis. J Dent Res 1992;8:263-71.  Back to cited text no. 4      
5.Cowman MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12:564-82.  Back to cited text no. 5      
6.Burneti RW, Haber MH, Hackel E, Hanson CA, Keren DF. Lee Lewandrowski E. Clinical laboratory medicine. Philadelphia, U.S: Williams and Wilkins; 1994. p. 1113-20.  Back to cited text no. 6      
7.Perez C, Pauli M, Bazerque P. An antibiotic assay by the well agar diffusion method. Acta Biologiae Et Medicine Experiments 1990;15:113-5.  Back to cited text no. 7      
8.Ahmad I, Mehmood Z, Mohammad F. Screening of some Indian medicinal plants for their antimicrobial properties. J Ethanopharmacol 1998;62:183-93.   Back to cited text no. 8      
9.Prabhat , Navneet, Shrikrishna. Antibacterial activity of Apamarga (Achyranthes aspera). National Academy of Science Letters 2005;28:379-81.  Back to cited text no. 9      
10.Indian Pharmacopoeia. The controller of publication, Vol. 2. Delhi: (CSIR); 1996: p. A-100-7.  Back to cited text no. 10      
11.Basri DF, Fan S. The potential of aqueous and acetone extract of galls of Q. infectoria as antimicrobial agent. Indian J of Pharmacol 2005;37:26-9.  Back to cited text no. 11      
12.Voravuthikunchai SP, Kitpipit L. Activity of medicinal plant extracts against hospital isolates of methicillin-resistant S. aureus. Clin Microbiol Infect 2005;11:510-2.   Back to cited text no. 12      
13.Chusri S, Voravuthikunchai SP. Q.infectoria: A candidate for the control of methicillin-resistant S. aureus infections. Phytother Res 2008;22:560-2.   Back to cited text no. 13      
14.Prabhat , Navneet, Shrikrishna. Antibacterial activity of Bakula (Mimusops elenigi). Environmental Conservation Journal 2005;3:59-61.  Back to cited text no. 14      
15.Ikram M, Nowshad F. Constituents of Quercus infectoria. Planta Medica 1977;31:286-7.  Back to cited text no. 15  [PUBMED]  [FULLTEXT]  
16.Leach CK. The phenolic contents of some British cynipid galls. Cecidology 1986;1:10-2.  Back to cited text no. 16      
17.Irobi ON, Moo-Young M, Anderson WA, Daramola SO. Antibacterial activity of bark extracts of Bridelia ferruginea (Euphorbiaceae). J of Ethanopharmacol 1994;43:185-90.  Back to cited text no. 17      
18.Djipa CD, Delmee M, Quetin-Leclercq J. Antimicrobial activity of bark extracts of Syzygium jambos (L.) Alston (Myrtaceae). J of Ethanopharmacol 2000;71:307-13.  Back to cited text no. 18      
19.Kottakkal AV. Indian Medicinal Plants. Vol. 4. Orient Longman Ltd; 1995.   Back to cited text no. 19      
20.Bhattacharjee SK. Handbook of medicinal plants. India: Pointer Publishers; 2001.  Back to cited text no. 20      

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Correspondence Address:
Archa Vermani
Department of Botany and Microbiology, Gurukul Kangri University, Haridwar
India
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DOI: 10.4103/0970-9290.57380

PMID: 19884719

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    Tables

  [Table 1], [Table 2]

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