ANTIBACTERIAL ACTIVITY OF ANNONA SQUAMOSA L. AND ANNONA RETICULATA L. AGAINST CLINICAL ISOLATES OF MUTANS STREPTOCOCCI THE CAUSATIVE AGENTS OF DENTAL CARIES
Objective: Evaluation of antibacterial activity of Annona squamosa L and Annona reticulata L against Streptococcus mutans and Streptococcus sobrinus
(mutans streptococci MS) the causative agent of dental caries.
Methods: Leaves and bark of A. squamosa and A. reticulata were collected and extracted by methanol solvent using soxhlet apparatus. Clinical samples
were isolated from dental caries subjects, and identified by 16S rDNA sequencing. The isolates were tested to study the antibacterial activity of
A. squamosa and A. reticulata leaves and bark by disc diffusion method. 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, and
100 mg concentration of the extract were used to study the antibacteial activity of S. mutans and S. sobrinus. Ampicillin (AMP) 10 Î¼g was used as a
positive control in all the species.
Results: The isolates were identified based on 16S rDNA sequencing as two Streptococcus mutans and two Streptococcus sobrinus. A. squamosa bark
showed antibacterial activity against MS species, while, at 100 mg concentration leaves extract of A. squamosa did not show any inhibition against
MS species. However, no inhibitory activity being shown in A. reticulata (leaves and bark) against MS species. The result indicates that S. sobrinus is
more susceptible to the A. squamosa bark extract than S. mutans. Not much difference showed between the species in regards of different concentration.
Conclusion: A. squamosa bark showed antibacterial activity against MS species, further investigation of the A. squamosa bark should be studied to
determine the anti-caries properties.
Keywords: Annona squamosa, Annona reticulata, Plant extract, Dental caries, Mutans Streptococci, Antibacterial susceptibility testing,
16S rDNA sequencing.
1. Ramos-Gomez FJ, Weintraub JA, Gansky SA, Hoover CI, Featherstone JD.
Bacterial, behavioral and environmental factors associated with early childhood caries. J Clin Pediatr Dent 2002;26(2):165-73.
2. Loesche WI. Role of Streptococcus mutans in human dental decay. Microbiol Rev 1986;50(4):353-80.
3. Hirose H, Hirose K, Isogai E, Miura H, Ueda I. Close association between Streptococcus sobrinus in the saliva of young children and smooth-surface caries increment. Caries Res 1993;27(4):292-7.
4. World Intellectual Property Organization. Herbal Formulation Capable of Preventing Alcohol-induced Hangover, Methods of Preparing the Same and use Thereof. Wo 2006;131932.
5. Morton J, editor. Sugar apple. Fruits of Warm Climate. Miami: Greensboro Media; 1987. p. 69-72.
6. Nirmal SA, Gaikwad SB, Dhasade VV, Dhikale RS, Kotkar PV, Dighe SS. Anthelmintic activity of Annona reticulata leaves. Res J Pharm Biol Chem Sci 2010;1(1):115-8.
7. Bhalke RD, Chavan MJ. Analgesic and CNS depressant activities of extracts of Annona reticulate Linn. bark. Phytopharmacology 2011;1(5):160-5.
8. Rahman SM, Rashedul MI, Rahman S, Mosaiab T, Ahmed R, Khatun F, et al. Antihyperglycemic studies with methanol extract of Annona reticulata L. (Annonaceae) and Carissa carandas L. (Apocynaceae) leaves in Swiss albino mice. Adv Nat Appl Sci 2011;5(2):218-22.
9. Shirwaikar A, Rajendran K, Kumar CD. In vitro antioxidant studies of Annona squamosa Linn leaves. Indian J Exp Biol 2004;42(8):803-7.
10. Vohora SB, Kumar I, Naqvi SA. Phytochemical, pharmacological, antibacterial and anti-ovulatory studies on Annona squamosa. Planta Med 1975;28(1):97-100.
11. Asolkar LV, Kakkar KK, Chakre OJ. Glossary of Indian Medicinal Plants with Active Principles. New Delhi: Publication and Information Directorate; 1992. p. 72-3.
12. Yoganarashimhan SN. Medicinal Plants of India. Vol II. TamilNadu, Bangalore: Cyber Media; 2000. p. 71.
13. Patel JD, Kumar V. Annona squamosa L.: Phytochemical analysis and antimicrobial screening. J Pharm Res 2008;1(1):34-8.
14. Saha R. Pharmacognosy and pharmacology of Annona squamosa: A review. Int J Pharm Life Sci 2011;2(10):1183-9.
15. Chavan MJ, Shinde DB, Nirmal SA. Major volatile constituent of Annona squamosa L. bark. Nat Prod Res 2006;20(8):754-7.
16. Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem 1975;250(10):4007-12.
17. Pardhasaradhi BV, Reddy M, Ali AM, Kumari AL, Khar A. Differential cytotoxic effects of Annona squamosa seed extracts on human tumour cell linesole of reactive oxygen species and glutathione. J Biosci 2005;30(2):237-44.
18. Kachhawa JB, Sharma N, Tyagi S, Sharma KK. Screening of stem bark methanol extract of Annona squamosa for antibacterial activity. Int J Curr Pharm Res 2012;4(1):48-50.
19. Padhi LP, Panda SK, Satapathy SN, Dutta SK. In vitro evaluation of antibacterial potential of Annona squamosa L. and Annona reticulata L. from similipal biosphere reserve, Orissa, India. J Agric Technol 2011;7(1):133-42.
20. Dhamodhar P, Sreenivasa Murthy, Channarayappa, Shanthakumar SS, Indiresha HN. Prevalence, characterization and heterogeneity studies on Streptococcus mutans isolated from Bangalore urban population. Int J Pharm Bio Sci, 2014; 5(3):122-8.
21. Ito K, Nakamura Y, Tokunaga T, Iijima D, Fukushima K. Anti-cariogenic properties of a water-soluble extract from cacao. Biosci Biotechnol Biochem 2003;67(12):2567-73.
22. Hwang JK, Chung JY, Baek NI, Park JH. Isopanduratin A from Kaempferia pandurata as an active antibacterial agent against cariogenic Streptococcus mutans. Int J Antimicrob Agents 2004;23(4):377-81.
23. Moreira M, Noschang J, Neiva IF, Carvalho Y, Vicente VA. Methodological variations in the isolation of genomic DNA from Streptococcus bacteria. Braz Arch Biol Technol 2010;53(4):845-9.
24. Lim SH, Seo JS, Yoon YJ, Kim KW, Yoo SY, Kim HS, et al. Effect of leaf-extract from Camellia sinensis and seed-extract from Casia tora on the viability of MS isolated from the interface between orthodontic brackets and tooth surfaces. Korean J Orthod 2003;33:381-9.
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