& Bioch ial em b ic ro a c l i T M e c Endang et al., J Microb Biochem Technol 2017, 9:6 f h o Journal of n l o a n l DOI: 10.4172/1948-5948.1000384 o r g u y o J ISSN: 1948-5948 Microbial & Biochemical Technology Review Article Open Access Antibacterial Activity toward Streptococcus mutans and Antioxidant from Traditional Betel Chew Formulation of Indonesia Endang Verawati1, Tri Wardani Widowati2, Budi Santoso2, Siti Rusdiana Puspa Dewi2 and Rindit Pambayun2 1Post Graduate Student Student of Postgraduate School Program, Faculty of Agriculture, Sriwijaya University, Indonesia 2Lecturer of Postgraduate School Program, Faculty of Agriculture, Sriwijaya University, Indonesia Abstract The objective of this research was to determine antibacterial activity toward Streptococcus mutans and antioxidant. Non-Factorial Randomized Block Design method was used in this study. The first stage used non- factorial randomized block design which consisted of three blocks and six treatments as follows: F1 (8 g betel leaf, 2 g betel lime), F2 (8 g betel leaf, 2 g betel lime, 2 g areca nut, 1 g gambier), F3 (8 g betel leaf, 2 g betel lime, 2.5 g areca nut, 1.5 g gambier), F4 (8 g betel leaf, 2 g betel lime, 3 g areca nut, 2 g gambier), F5 (8 g betel leaf, 2 g betel lime , 3.5 g areca nut, 2.5 g gambier) and F6 (cefadroxil).The observed parameters in betel chew formulation were antibacterial activity, cellulair metabolites leakage and antioxidant. Results of chemical and microbiological analyses showed that the best treatment was found on F5 treatment (8 g betel leaf, 2 g betel lime, 3.5 g areca nut, 2.5 g gambier) antibacterial activity of 8.25 mm, with antioxidant IC50 of 2.77 mg/ml and cellulair metabolites leakage of 1.22 nm (at wave length of 260 nm) and 1.51 nm (at wave length of 280 nm), respectively. Keywords: Betel chew extract; Antibacterial; Antioxidant; Gambier can be used as growth inhibitor for bacteria of Streptococcus Streptococcus mutans mutans [9]. Betel lime has chemical formula of CaOH2 (calcium hydroxide) with calcium as its main component so that it can be used to strengthen the Introduction teeth [10]. Betel chew is mixture of substances such as betel leaf, areca nut, lime Formulation used in betel chew activity has antimicrobial and and gambier used in betel chew activity. antioxidant compounds. Antimicrobia especially bacteria found in According to Flora et al. [1], results of research done in England teeth is Streptococcus mutans. Antimicrobia is compound that capable by South Asia Immigrants showed that chewing of betel leaf and areca to inhibit the growth of microorganisms [11]. Streptococcus mutans nut gave freshness feeling, reducing stress and strengthen the teeth. is bacteria found in teeth and one of disease caused by Streptococcus Research results done by Parianti and Ariyasa [2] by using elderly mutans is dental caries [12]. Dental caries is infectional disease that respondents having age of 60 to 89 years which consisted of 37 betel destroys teeth structure (enamel). This disease can cause teeth cavity chewers at Batubulan Kangin Village, Sukawati sub district, Gianyar [13]. Antioxidant is a compound that protects cells from damaging due district, showed that the longer the betel chew activity by respondents, to unstable compound called free radical [14]. the less numbers of dental caries experienced by respondents. The Betel chew activity has very beneficial effect on health and teeth. research results in term of the effect of betel chew activity time on dental Therefore, identification of antibacterial activity toward Streptococcus caries level can be seen in Table 1. mutans as well as antioxidant was done in term of betel chew activity Mixed materials for betel chew activity are betel leaf, areca nut, lime and that has very beneficial effect on health and teeth. gambier that have different nutrient content. Betel leaf is one of traditional The main objectives of this research are as follows to determine the medicinal plants having nutrient content as follows: water content of 85.14%, addition of areca nut extract formulation that capable to inhibit the protein of 3.1%, lipid of 0.8%, carbohydrate of 6.1%, fibre of 2.4%, calcium growth Streptococcus mutans bacteria. To determine the addition of of 230 mg, phosphor of 40 mg, iron of 7 mg, iron ion of 3.5 mg, carotene areca nut extract formulation that capable to be used as antioxidant. of 96000 IU, tamin of 70 mg, riboflavin of 30 mg, nicotinate acid of 0.7 mg, vitamin C of 5 mg, tannins of 1.3%, essential oil of 45% with bethel phenol Materials and Methods as its main component [3,4]. Betel leaf contains phenol that has role as toxic Design experiment for microbia by inhibiting enzyme activity from disturber microbial [5]. Nutrient content of areca nut are carbohydrate, lipid, fibre, polyphenol and Design experiment used in this research was Non-Factorial alkaloid 0.3 to 0.6%, tanin of 15% and fat of 14% [6]. According to Mamonto et al. [7], areca nut can be used as antioxidant. Nutrient content of gambier are catechin of 7 to 33%, catechu tannic acid of 20 to 55%, pyrocatecol of 20 *Corresponding author: Endang V, Postgraduate school program, Faculty of to 30%, fluorescence gambier of 1to 3%, red catechu of 3 to 5%, quersetin agriculture, Sriwijaya University Palembang, South Sumatra, Indonesia, Tel: of 2 to 4%, fixed oil of 1 to 2%, wax of 1% to 2% and alkaloid <1% [8]. 62711580068; Fax: 62711 580068; E-mail: [email protected] Received November 23, 2017; Accepted December 21, 2017; Published December 28, 2017 Betel chew Category of dental caries Citation: Endang V, Tri Wardani W, Budi S, Siti Rusdiaana Puspa D, Rindit P (2017) activity time Very low Low Medium High Total Antibacterial Activity toward Streptococcus mutans and Antioxidant from Traditional 1 to 5 years 0 1 14 3 18 Betel Chew Formulation of Indonesia. J Microb Biochem Technol 9: 316-320. doi: 6 to 10 years 5 1 1 3 10 10.4172/1948-5948.1000384 >10 years 9 0 0 0 9 Copyright: © 2017 Endang V, et al. This is an open-access article distributed under Total 14 2 15 6 37 the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and Table 1: The effect of betel chews activity time on dental caries level. source are credited. J Microb Biochem Technol, an open access journal ISSN: 1948-5948 Volume 9(6): 316-320 (2017) - 316 Citation: Endang V, Tri Wardani W, Budi S, Siti Rusdiaana Puspa D, Rindit P (2017) Antibacterial Activity toward Streptococcus mutans and Antioxidant from Traditional Betel Chew Formulation of Indonesia. J Microb Biochem Technol 9: 316-320. doi: 10.4172/1948-5948.1000384 Materials F1 F2 F3 F4 F5 F6 volume of 10 Ml; Betel leaf (g) 8 8 8 8 8 0 2. Five dilution series are made from this sample which added Betel lime (g) 2 2 2 2 2 0 with methanol until the volume is 5 mL, i.e., dilution of 0x, 0.2x, Areca nut (g) 0 2 2.5 3 3.5 0 0.4x, 0.6x, 0.8x and 1x; Gambier (g) 0 1 1.5 2 2.5 0 Cefadroxil (g) 0 0 0 0 0 0.5 3. Diluted sample of 2 mL is taken and added with 2 mL DPPH solution; Table 2: Formulation of betel chew extract. 4. Sample is subjected to vortex treatment until homogenous; Randomized Block Design. Betel chews formulation development process. Treatments level can be seen in Table 2. Betel chew formulation 5. Sample is incubated at 37°C for 30 min; is obtained from survey results related to people who used to do betel 6. Absorbency of sample is determined by using spectrophotometer chew activity. Treatment for betel chew formulation was increased four- at λ=517 nm; fold from the survey results value because this formulation was not indicating antibacterial capability during preliminary study. Inhibition Percentage (%)= x100% Research procedure Remarks: This research consisted of jelly candy processing procedure Ablank: Result of spectrophotometer reading at 0 min. according to Rahmi et al. that had been modified as follows [15]: Asample: Result of spectrophotometer reading at 30 min. 1. Gambier is mashed and weighed according to treatments, i.e., After inhibition percentage value is obtained, then IC 50 value 1 g, 1.5 g, 2 g, 2.5 g. can be calculated by developing the linear equation from dilution 2. Areca nut seed is mashed and weighed according to treatments, concentration and inhibition percentage. Nilai IC50 value is obtained i.e., 2 g, 2.5 g, 3 g, 3.5 g. from X values of the above linear equation and Y value is 50. The linear equation of dilution concentration (as absica values) and inhibition 3. Supplementary ingredients for betel chew are weighed, i.e., 2g percentage (as ordinate values) can be written as follows: betel lime and 8 g betel leaves. y=ax+b 4. All weighed ingredients are subsequently blended and water is added up to 40 mL until homogenous and had dark red color. Test of cellular metabolite leakage 5. The blended ingredients are subsequently macerated for 24 h at The test of cellular metabolite leakage according to Chia et al. is room temperature. After 24 h, solution is separated (filtrated) done by using spectrophotometer and absorbency measurement is by using saring paper. conducted at 260 nm wave length (for nitrogen content of nucleate acid) and 280 nm wave length (for determination of nitrogen content Test of antibacterial activity of protein) [19].