{"id":56404,"date":"2024-03-20T11:06:08","date_gmt":"2024-03-20T11:06:08","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=56404"},"modified":"2024-04-01T19:29:04","modified_gmt":"2024-04-01T19:29:04","slug":"antibacterial-and-antibiofilm-activity-of-ethanol-extract-of-batak-onion-bulbs-allium-chinense-g-don-against-streptococcus-mutans-and-enterococcus-faecalis","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no1\/antibacterial-and-antibiofilm-activity-of-ethanol-extract-of-batak-onion-bulbs-allium-chinense-g-don-against-streptococcus-mutans-and-enterococcus-faecalis\/","title":{"rendered":"Antibacterial and Antibiofilm Activity of Ethanol Extract of Batak Onion Bulbs (Allium chinense G.Don.) against Streptococcus mutans and Enterococcus faecalis."},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Indonesia is an agricultural country with more than 30,000 plant species, and about 7,000 of them have been recorded as medicinal plants.1<\/sup> Allium genus plants are well known and are often used by the public as a spice and traditional medicine. One of the Allium genus plants that has been widely utilized by the Indonesian people, especially the Batak tribe, is Batak Onion (Allium chinense<\/em> G.Don.).2<\/sup><\/p>\n\n\n\n

Batak\nonions or chives can grow up to 50 cm in height with slender, bright green,\nthin-walled, 3-5 angled leaves (not round),  are not too stiff upright, and have\nlavender-colored flowers with long stalks and stamens sticking out. Batak\nonions grow in clumps and form many bulbs with a solid stalk and a white color.\nBatak onion bulbs have a diameter of 4-5 cm, are grayish-white to purple, are wrapped in transparent skin, and are\nwhite.3,4<\/sup><\/p>\n\n\n\n

Allium\ngenus plants\nhave known antimicrobial compounds such as allicin<\/em>,\ndialyl disulfide<\/em>, ajoene<\/em>, and 3-(allyltrisulfanyl)-2-amino-propanoic acid<\/em>.Based on\nHarborne’s phytochemical identification, 70% of Batak onion bulbs’\nethanol extract has secondary metabolite compounds of alkaloids, flavonoids, and\nsaponins.2,5<\/sup><\/p>\n\n\n\n

Dental\ncaries is one of the most common dental health problems experienced by children\nand adults.6<\/sup> This disease occurs due to the interacting processes of\nteeth, saliva (the host), microorganisms, substrate, and time. Although the\ncause of caries is multifactorial, the main trigger is the dominant bacteria Streptococcus mutans <\/em> (S.mutans<\/em>).7<\/sup> Caries\nthat are not treated immediately will cause severity, so a root canal or endodontic treatment\nis needed to handle it. In endodontic treatment, root canal infections usually\noccur. This infection is usually caused by microorganisms, especially Enterococcus<\/em> faecalis <\/em>(E.faecalis<\/em>).8<\/sup><\/p>\n\n\n\n

Chlorhexidine\nis an antimicrobial agent effective as a mouthwash in preventing the buildup of\ncaries-causing bacterial plaque and as a root canal irrigation agent in\nendodontic treatment.9,10<\/sup> Although superior, chlorhexidine has the\ndisadvantage of causing discoloration of the teeth when used continuously as a\nmouthwash and cannot be used as a single root canal irrigation material, so it\nmust be combined with other irrigation solutions. Therefore, other alternative\nmaterials are needed to replace chlorhexidine.9,11<\/sup><\/p>\n\n\n\n

The\nstudy conducted by another researcher demonstrated that the ethanol\nextract from Allium chinense exhibited significant antioxidant activity.\nAdditionally, the high-content essential oil extract of the plant significantly\ndecreased serum and hepatic levels of total cholesterol.12 <\/sup>Research shows that Batak onion bulb extract\nhas an antimicrobial response in inhibiting the growth of Escherichia coli<\/em>, Salmonella\ntyphi<\/em>, Staphylococcus aureus<\/em>, Bacillus subtilis<\/em> and Candida albicans<\/em>.2<\/sup> Other\nreports showed that\nBatak union bulb extract can inhibit the growth of Methicillin-Resistant\nStaphylococcus aureus<\/em> (MRSA).13<\/sup> Many articles are available for onion studies as\nantibacterial and antibiolfilm activities. One study that showed Onion (Allium cepa<\/em>\nL.) had potential antibacterial and antibiofilm activity against Escherichia\ncoli<\/em>, Pseudomonas aeruginosa<\/em>, Staphylococcus aureus<\/em>, and Bacillus\ncereus<\/em>. No research reports on the antibacterial and antibiofilm\nactivity possessed by Batak onion bulb extract against S.mutans<\/em> and E.faecalis<\/em>.14<\/sup>\nThus, we are interested in conducting antibacterial activity test research\nbased on minimum inhibitory concentration (MIC), minimum bactericidal\nconcentration (MBC), and antibiofilm of 70% ethanol extract of Batak onion\nbulbs against S.mutans<\/em> and E.faecalis<\/em>.<\/p>\n\n\n\n

Materials and Methods<\/strong><\/p>\n\n\n\n

This\nlaboratory experimental study has a post-test only with a controlled\ngroup design. The research samples used in this study were Streptococcus mutans<\/em> ATCC\u00ae 25175TM and Enterococcus faecalis<\/em> ATCC\u00ae 29212TM, which had been subcultured and\nisolated at the Universitas Sumatera Utara Hospital Clinical Microbiology Unit,\n70% ethanol of Batak onion bulbs with concentrations of 50%; 25%; 12.5%; 6.25%;\n3.125%; 1.56%; and 0.78%, and a control group consisting of chlorhexidine 0.2%\nas a positive control and Dimethyl Sulfoxide (DMSO) as a negative control. All\nsamples were treated three times, so the sample size of this study was 54\nsamples consisting of 27 samples for S.mutans<\/em>\nand 27 samples for E.faecalis<\/em>.<\/p>\n\n\n\n

Extract preparation<\/strong><\/p>\n\n\n\n

Preparation\nof the extract begins with making simplisia from 1 kg of Batak onions from\nBrastagi District. Batak onion bulbs are cleaned in running water, cut, sliced\nwith a slicing knife, and dried in an oven at 25\u00b0 for 25 hours until completely\ndry, with characteristic until the water content is less than 10% and easily broken\nif kneaded.15<\/sup> The dried bulbs were crushed with a porcelain pestle\nand pulverized with a blender (Cosmos CB 282P\u00ae<\/sup>), weighted and\nexpected to produce a fine powder of as much as 150 grams, then put in an airtight container. <\/p>\n\n\n\n

The\nextraction of simplisia is done by maceration. 150 grams of simplisia that have\nbeen finely added with 1 liter of 70% ethanol in the vessel then stirred manually\nwith a stick under atmospheric pressure for the first 6 hours and allowed to\nstand for 18 hours with the vessel closed while occasionally stirring. In the\nnext step, the marinade is filtered to obtain filtrate (Maserat I), and the\ndregs are collected and accommodated in a white plastic bottle container. Did the\nsame extraction of the dregs in 500 ml of 70% at room temperature for 24 hours while\nstirring several times, and then filtered and collected them (Maserat II). All\nfiltrates obtained were combined and evaporated with a rotary evaporator RE-2010 at 40\u00b0C with 80 rpm\nuntil a thick extract was obtained.<\/p>\n\n\n\n

Before\nthe antibacterial test, screening was carried out for the phytochemicals in the\n70% ethanol extract of Batak onion bulbs. Furthermore, the concentration of the\ntest solution was made by diluting DMSO against the thick extract of Batak\nonion bulbs to obtain extract concentrations of 50%, 25%, 12.5%, 6.25%, 3.125%,\n1.56%,\nand 0.78%.<\/p>\n\n\n\n

MIC Testing<\/strong><\/p>\n\n\n\n

Mueller\nHinton Agar media (Himedia<\/em>) is made\nby dissolving 38 grams of powder with 1 liter of distilled water and then\nsterilized in an autoclave at 121\u00b0C for 15 minutes and then poured into petri\ndishes. Mueller Hinton Broth media (Himedia) is made by dissolving 21 grams of\npowder with 1 liter of distilled water, then sterilized in an autoclave at\n121\u00b0C for 15 minutes, and then poured into Petri dishes.<\/p>\n\n\n\n

Making\nbacterial suspensions is done by taking one dose of S.mutans<\/em> and E.faecalis<\/em>\ncolonies, then each is dissolved in a test tube containing 10 ml of 0.45% NaCl,\nhomogenized with a vortex mixer, and incubated for 1-2 hours so that a\nMcFarland Standard turbidity of 0.5 is obtained. Perform dilutions of each\nbacterium by mixing 0.1 ml of suspension and 9.9 ml of MHB and homogenizing\nagain with a vortex mixer.<\/p>\n\n\n\n

Measurement\nof antibacterial activity based on the diameter of the inhibition zone was\ncarried out by the Kirby-Baurer disc diffusion method. Paper discs dripped with\nextracts according to the total sample were placed on Petri dishes containing\nMHA and bacteria that had been homogenized and then incubated for 24 hours.\nAfter incubation, observations were made by measuring the clear zone using\ncalipers. The smallest concentration that has a clear zone is designated as the\nMIC.<\/p>\n\n\n\n

MBC Testing<\/strong><\/p>\n\n\n\n

The\nDetermination of MBC is done by counting the number of bacterial colonies. The\nclear zone of MIC results from each bacterium was streaked with a sterile\ncotton swab and then dipped in a test tube containing MHB and allowed to stand\nfor 10 minutes. 1 ml of MHB was taken from the test tube, dripped on sterile\npetri, and added to PCA. It was then homogenized and incubated in an incubator\nat 37\u00b0C for 24 hours, after which bacterial colonies were counted with a colony\ncounter. The concentration that can reduce bacteria by 98-99% of the initial\nnumber of colonies is determined as MBC.<\/p>\n\n\n\n

Antibiofilm Testing<\/strong><\/p>\n\n\n\n

Antibiofilm\ntesting used the Static Microtiter Plate Assay method by culturing each\nbacterial suspension in Trypticase Soy Broth (TSB) + 1% Glucose. 100\u00b5L of a 70%\nethanol extract solution of various concentrations of Batak onion bulbs and\n100\u00b5L of each bacterial suspension was put into a 96-well microplate and\nincubated at 37\u00b0C for 48 hours. After incubation, the contents of the\nmicroplate were removed, rinsed with Phosphate Buffered Saline (PBS) and fixed\nwith a bunsen. Crystal violet solution (1% w\/v) was given to each microplate\ncontainer of 200 \u00b5L, incubated for 15 minutes at room temperature, and then\nrinsed with PBS. After drying, enter 200 \u00b5L of 96% ethanol in each wheel and\nincubate again. The microplate is then inserted into the microplate reader with\na wavelength of 600nm. Repeat three times, and then the percentage is\ncalculated according to Costa et al. (2016):12 <\/sup><\/p>\n\n\n\n

Test Analysis<\/strong><\/p>\n\n\n\n

The collected data were statistically\nexamined With the aid of Statistical Product Service Solutions (SPSS). The Shapiro-Wilk method was used\nto evaluate the normality of the data, and one-way ANOVA and LSD post hoc tests\nwith a significance threshold of p<0.05 were used to continue the data\nanalysis.<\/p>\n\n\n\n

Result<\/strong><\/p>\n\n\n\n

According\nto the results of phytochemical screening, the secondary metabolites contained\nin the 70% ethanol extract of Batak onion bulbs are flavonoids, alkaloids,\nsaponins and glycosides.<\/p>\n\n\n\n

Antibacterial\ntesting based on the diameter of the inhibition zone obtained MIC at a\nconcentration of 0.78% with an average inhibition zone of S.mutans<\/em> 9.00\u00b10.43 mm and E.<\/em>\nfaecalis<\/em> 8.06\u00b10.20 mm (Table 1).  <\/p>\n\n\n\n

From\nthe measurement of bacterial colonies, no MBC was found because all test\nconcentrations could not reduce bacteria by 98-99%. The largest percentage of\nbacterial reduction produced was in the 50% extract with a percentage reduction\nof 94.67% in S.mutans<\/em> and 84.19% in E.faecalis<\/em> (Table 2). <\/p>\n\n\n\n

Antibiofilm\nmeasurements obtained the results that 70% ethanol extract of Batak onion bulbs\nat a concentration of 0.78% was able to form antibiofilm against S.mutans<\/em> and E.faecalis<\/em> with a percentage of 84.55% and 85.73% (Table 3). This\nantibiofilm increased as the concentration increased to 12.5% and decreased\nantibiofilm reduction at 25% and 50% concentrations.<\/p>\n\n\n\n

In\nTable 4, the results from the Post Hoc Test (LSD) show a comparison of\nconcentrations that have significant differences from 70% ethanol extract of\nBatak onion bulbs against S.mutans<\/em>\n(p<0.05). Post hoc inhibition of S.mutans<\/em>\ngroup (I) all have significant differences except at concentrations of 12.5%\nagainst control (+), 6.25% against control (+), 3.125% against 1.56%, and 1.56%\nagainst 0.78%. Almost all post-hoc\ntest results of MBC (II) had significant differences, except at concentrations\nof 50% against 25% and 50% against the control (+). The antibiofilm group (III)\nalmost all had significant differences, except at concentrations of 50% against\n25%,\n50% against 0.78%, 25% against 1.56%; 25% against 0.78%; 12.5% against 6.25%;\n12.5% against 3.125%; 12.5% against 1.56%; 6.25% against 3.125%; 6.25% against\n1.56%; 3.125%against 1.56%: and 1.56% against 0.78%.<\/p>\n\n\n\n

In Table 5, the results from the post hoc test (LSD) show a concentration comparison that has a significant difference from 70% ethanol extract of Batak onion bulbs against E.faecalis<\/em> (p<0.05). Post hoc group inhibition zone (I) against E.faecalis<\/em> mostly has significant differences, except at concentrations of 6.25% against 3.125% and 3.125% against 1.56%. In the post hoc test of MBC (II), all of them have significantly different concentration comparisons (p<0.05). Almost all concentration comparisons in the post hoc antibiofilm group (III) have significant differences except in the concentration group against the control group.<\/p>\n\n\n\n

Table 1: Antibacterial activity based on the inhibition zone diameter of 70% ethanol extract of Batak onion bulbs against Streptococcus mutans<\/em> and Enterococcus<\/em> faecalis<\/em><\/strong>.<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Treatment Group<\/strong><\/p>\n<\/td>\n

\n

Streptococcus mutans<\/em><\/strong><\/p>\n<\/td>\n

\n

Enterococcus faecalis<\/em><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Mean diameter of inhibition zone (mm)<\/strong><\/p>\n<\/td>\n

\n

p<\/strong><\/p>\n<\/td>\n

\n

Mean diameter of inhibition zone (mm)<\/strong><\/p>\n<\/td>\n

\n

p<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

50%<\/p>\n<\/td>\n

\n

24.66\u00b10.75<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

13.86\u00b10.55<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

25%<\/p>\n<\/td>\n

\n

20.83\u00b11.36<\/p>\n<\/td>\n

\n

12.56\u00b10.41<\/p>\n<\/td>\n<\/tr>\n

\n

12.5%<\/p>\n<\/td>\n

\n

13.53\u00b10.50<\/p>\n<\/td>\n

\n

11.16\u00b10.87<\/p>\n<\/td>\n<\/tr>\n

\n

6.25%<\/p>\n<\/td>\n

\n

12.40\u00b10.36<\/p>\n<\/td>\n

\n

10.03\u00b10.60<\/p>\n<\/td>\n<\/tr>\n

\n

3.125%<\/p>\n<\/td>\n

\n

10.13\u00b10.30<\/p>\n<\/td>\n

\n

9.53\u00b10.35<\/p>\n<\/td>\n<\/tr>\n

\n

1.56%<\/p>\n<\/td>\n

\n

9.76\u00b10.47<\/p>\n<\/td>\n

\n

8.96\u00b10.32<\/p>\n<\/td>\n<\/tr>\n

\n

0.78%<\/p>\n<\/td>\n

\n

9.00\u00b10.43<\/p>\n<\/td>\n

\n

8.06\u00b10.20<\/p>\n<\/td>\n<\/tr>\n

\n

Chlorhexidine <\/em>0.2%<\/p>\n<\/td>\n

\n

12.8\u00b10.36<\/p>\n<\/td>\n

\n

17.90\u00b10.30<\/p>\n<\/td>\n<\/tr>\n

\n

DMSO<\/p>\n<\/td>\n

\n

0.00\u00b10.00<\/p>\n<\/td>\n

\n

0.00\u00b10.00<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

     *Statistically significant (p<0.05)<\/p>\n\n\n

Table 2: Antibacterial activity based on bactericidal activity of 70% ethanol extract of Batak   onion bulbs against Streptococcus mutans<\/em> and Enterococcus<\/em> faecalis<\/em><\/strong>.<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Treatment Group<\/strong><\/p>\n<\/td>\n

\n

Streptococcus mutans<\/em><\/strong><\/p>\n<\/td>\n

\n

Enterococcus facealis<\/em><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Number of colonies (X\u0305\u00b1SD)<\/strong><\/p>\n

(CFU\/ml)<\/strong><\/p>\n<\/td>\n

\n

% Reduction<\/strong><\/p>\n<\/td>\n

\n

p<\/strong><\/p>\n<\/td>\n

\n

Number of colonies (X\u0305\u00b1SD)<\/strong><\/p>\n

(CFU\/ml)<\/strong><\/p>\n<\/td>\n

\n

% Reduction<\/strong><\/p>\n<\/td>\n

\n

p<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

50%<\/p>\n<\/td>\n

\n

67\u00b112.85<\/p>\n<\/td>\n

\n

94.67%<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

182\u00b13.78<\/p>\n<\/td>\n

\n

84.19%<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

25%<\/p>\n<\/td>\n

\n

117\u00b14.72<\/p>\n<\/td>\n

\n

90.67%<\/p>\n<\/td>\n

\n

290\u00b16.55<\/p>\n<\/td>\n

\n

74.80%<\/p>\n<\/td>\n<\/tr>\n

\n

12.5%<\/p>\n<\/td>\n

\n

213\u00b113.11<\/p>\n<\/td>\n

\n

83.05%<\/p>\n<\/td>\n

\n

371\u00b14.50<\/p>\n<\/td>\n

\n

67.77%<\/p>\n<\/td>\n<\/tr>\n

\n

6.25%<\/p>\n<\/td>\n

\n

309\u00b14.16<\/p>\n<\/td>\n

\n

75.42%<\/p>\n<\/td>\n

\n

484\u00b18.02<\/p>\n<\/td>\n

\n

57.95%<\/p>\n<\/td>\n<\/tr>\n

\n

3.125%<\/p>\n<\/td>\n

\n

406\u00b18.14<\/p>\n<\/td>\n

\n

67.70%<\/p>\n<\/td>\n

\n

571\u00b18.18<\/p>\n<\/td>\n

\n

50.39%<\/p>\n<\/td>\n<\/tr>\n

\n

1.56%<\/p>\n<\/td>\n

\n

522\u00b15.56<\/p>\n<\/td>\n

\n

58.47%<\/p>\n<\/td>\n

\n

662\u00b122.00<\/p>\n<\/td>\n

\n

42.48%<\/p>\n<\/td>\n<\/tr>\n

\n

0.78%<\/p>\n<\/td>\n

\n

641\u00b112.50<\/p>\n<\/td>\n

\n

49.00%<\/p>\n<\/td>\n

\n

755\u00b17.63<\/p>\n<\/td>\n

\n

34.40%<\/p>\n<\/td>\n<\/tr>\n

\n

Chlorhexidine <\/em>0.2%<\/p>\n<\/td>\n

\n

0\u00b10.00<\/p>\n<\/td>\n

\n

100.00%<\/p>\n<\/td>\n

\n

0\u00b10.00<\/p>\n<\/td>\n

\n

100%<\/p>\n<\/td>\n<\/tr>\n

\n

DMSO<\/em><\/p>\n<\/td>\n

\n

1257\u00b1156.61<\/p>\n<\/td>\n

\n

0%<\/p>\n<\/td>\n

\n

1151\u00b146.51<\/p>\n<\/td>\n

\n

0%<\/p>\n<\/td>\n<\/tr>\n<\/thead>\n<\/table>\n

*Statistically significant (p<0.05)<\/p>\n\n\n

Table 3: Antibiofilm activity of 70% ethanol extract of Batak onion bulbs against Streptococcus mutans<\/em> and Enterococcus<\/em> faecalis<\/em><\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Treatment Group<\/strong><\/p>\n<\/td>\n

\n

Streptococcus mutans<\/em><\/strong><\/p>\n<\/td>\n

\n

Enterococcus faecalis<\/em><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Average Optical Density <\/em>(OD)<\/strong><\/p>\n<\/td>\n

\n

% Reduction<\/strong><\/p>\n<\/td>\n

\n

p<\/strong><\/p>\n<\/td>\n

\n

Average Optical Density <\/em>(OD)<\/strong><\/p>\n<\/td>\n

\n

% Reduction<\/strong><\/p>\n<\/td>\n

\n

p<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

50%<\/p>\n<\/td>\n

\n

0.142\u00b10.009<\/p>\n<\/td>\n

\n

83.99%<\/p>\n<\/td>\n

\n

0.001*<\/p>\n<\/td>\n

\n

0.098\u00b10.006<\/p>\n<\/td>\n

\n

84.55%<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

25%<\/p>\n<\/td>\n

\n

0.136\u00b10.017<\/p>\n<\/td>\n

\n

84.66%<\/p>\n<\/td>\n

\n

0.091\u00b10.003<\/p>\n<\/td>\n

\n

86.02%<\/p>\n<\/td>\n<\/tr>\n

\n

12.5%<\/p>\n<\/td>\n

\n

0.116\u00b10.004<\/p>\n<\/td>\n

\n

86.92%<\/p>\n<\/td>\n

\n

0.086\u00b10.001<\/p>\n<\/td>\n

\n

86.60%<\/p>\n<\/td>\n<\/tr>\n

\n

6.25%<\/p>\n<\/td>\n

\n

0.118\u00b10.001<\/p>\n<\/td>\n

\n

86.69%<\/p>\n<\/td>\n

\n

0.088\u00b10.002<\/p>\n<\/td>\n

\n

87.19%<\/p>\n<\/td>\n<\/tr>\n

\n

3.125%<\/p>\n<\/td>\n

\n

0.121\u00b10.003<\/p>\n<\/td>\n

\n

86.35%<\/p>\n<\/td>\n

\n

0.092\u00b10.002<\/p>\n<\/td>\n

\n

87.48%<\/p>\n<\/td>\n<\/tr>\n

\n

1.56%<\/p>\n<\/td>\n

\n

0.124\u00b10.001<\/p>\n<\/td>\n

\n

86.02%<\/p>\n<\/td>\n

\n

0.097\u00b10.003<\/p>\n<\/td>\n

\n

86.75%<\/p>\n<\/td>\n<\/tr>\n

\n

0.78%<\/p>\n<\/td>\n

\n

0.137\u00b10.007<\/p>\n<\/td>\n

\n

84.55%<\/p>\n<\/td>\n

\n

0.105\u00b10.001<\/p>\n<\/td>\n

\n

85.73%<\/p>\n<\/td>\n<\/tr>\n

\n

Chlorhexidine <\/em>0.2%<\/p>\n<\/td>\n

\n

0.000\u00b10.000<\/p>\n<\/td>\n

\n

100,00%<\/p>\n<\/td>\n

\n

0,0000\u00b10,000<\/p>\n<\/td>\n

\n

100.00%<\/p>\n<\/td>\n<\/tr>\n

\n

DMSO<\/em><\/p>\n<\/td>\n

\n

0.887\u00b10.007<\/p>\n<\/td>\n

\n

0%<\/p>\n<\/td>\n

\n

0,6544\u00b10,063<\/p>\n<\/td>\n

\n

0%<\/p>\n<\/td>\n<\/tr>\n<\/thead>\n<\/table>\n

* Statistically significant (p<0.05)<\/p>\n\n\n

Table 4: LSD post hoc test results of differences in inhibition zone diameter, bactericidal, and antibiofilm activity test of 70% ethanol extract of Batak onion bulbs against Streptococcus mutans<\/em> <\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Concentration<\/strong><\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n<\/td>\n

\n

25%<\/strong><\/p>\n<\/td>\n

\n

12.5%<\/strong><\/p>\n<\/td>\n

\n

6.25%<\/strong><\/p>\n<\/td>\n

\n

3.125%<\/strong><\/p>\n<\/td>\n

\n

1.56%<\/strong><\/p>\n<\/td>\n

\n

0.78%<\/strong><\/p>\n<\/td>\n

\n

C(+)<\/strong><\/p>\n<\/td>\n

\n

C(-)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

50%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

0.265<\/p>\n<\/td>\n

\n

0.003*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.134<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

0.305<\/p>\n<\/td>\n

\n

0.001<\/p>\n<\/td>\n

\n

0.001*<\/p>\n<\/td>\n

\n

0.004<\/p>\n<\/td>\n

\n

0.010*<\/p>\n<\/td>\n

\n

0.389<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

25%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.040<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.014*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.007<\/p>\n<\/td>\n

\n

0.015<\/p>\n<\/td>\n

\n

0.042<\/p>\n<\/td>\n

\n

0.083<\/p>\n<\/td>\n

\n

0.866<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

12.5%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.038<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.164<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.038<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.743<\/p>\n<\/td>\n

\n

0.419<\/p>\n<\/td>\n

\n

0.251<\/p>\n<\/td>\n

\n

0.005*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

6.25%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.440<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.037<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.628<\/p>\n<\/td>\n

\n

0.405<\/p>\n<\/td>\n

\n

0.010*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

3.125%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.478<\/p>\n<\/td>\n

\n

0.038<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.016*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.724<\/p>\n<\/td>\n

\n

0.029<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

1.56%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.147<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.013<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.060<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

0.78%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

* Statistically significant (p<0.05)<\/p>\n

Description   I  : Inhibition zone diameter<\/p>\n

II : bactericidal activity<\/p>\n

III : Antibiofilm activity<\/p>\n\n\n

Table 5: LSD post hoc test results of differences in inhibition zone diameter, bactericidal, and antibiofilm activity test of 70% ethanol extract of Batak onion bulbs against Enterococcus<\/em> faecalis<\/em> <\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Concentration<\/strong><\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n<\/td>\n

\n

25%<\/strong><\/p>\n<\/td>\n

\n

12.5%<\/strong><\/p>\n<\/td>\n

\n

6.25%<\/strong><\/p>\n<\/td>\n

\n

3.125%<\/strong><\/p>\n<\/td>\n

\n

1.56%<\/strong><\/p>\n<\/td>\n

\n

0.78%<\/strong><\/p>\n<\/td>\n

\n

C(+)<\/strong><\/p>\n<\/td>\n

\n

C(-)<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

50%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

0.003*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

0.610<\/p>\n<\/td>\n

\n

0.403<\/p>\n<\/td>\n

\n

0.501<\/p>\n<\/td>\n

\n

0.691<\/p>\n<\/td>\n

\n

0.925<\/p>\n<\/td>\n

\n

0.627<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

25%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.002*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.741<\/p>\n<\/td>\n

\n

0.869<\/p>\n<\/td>\n

\n

0.909<\/p>\n<\/td>\n

\n

0.676<\/p>\n<\/td>\n

\n

0.324<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

12.5%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.008*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.867<\/p>\n<\/td>\n

\n

0.657<\/p>\n<\/td>\n

\n

0.457<\/p>\n<\/td>\n

\n

0.194<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

6.25%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.206<\/p>\n<\/td>\n

\n

0.012*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.780<\/p>\n<\/td>\n

\n

0.562<\/p>\n<\/td>\n

\n

0.253<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

3.125%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.155<\/p>\n<\/td>\n

\n

0.001*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.761<\/p>\n<\/td>\n

\n

0.381<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n

\n

0.000*<\/p>\n<\/td>\n<\/tr>\n

\n

1.56%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.030<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.563<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n<\/tr>\n

\n

0.78%<\/strong><\/p>\n<\/td>\n

\n

I<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n<\/tr>\n

\n

II<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n<\/tr>\n

\n

III<\/strong><\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n

\n

0.000<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

* Statistically significant (p<0.05) ;<\/p>\n

Description   I  : Inhibition zone diameter<\/p>\n

II : bactericidal activity<\/p>\n

III : Antibiofilm activity<\/p>\n\n\n

Discussion<\/strong><\/p>\n\n\n\n

70%\nethanol as a solvent was determined because ethanol is a universal solvent that\ncan dissolve all secondary metabolite compounds contained in simplicia and has\nlower toxicity than another organic solvent. 70% ethanol can dissolve phytochemical compounds more\noptimally because 70% ethanol still contains quite a lot of water (30%), which\nhelps the extraction process so that some of these compounds can\nbe attracted to ethanol, and some are attracted to water.16,17<\/sup><\/p>\n\n\n\n

The\nresults of the antibacterial activity test based on the diameter of the\ninhibition zone of 70% ethanol extract of Batak onion bulbs show that Batak\nonion bulbs have antibacterial activity (p<0.05) against S.mutans<\/em> and E.faecalis<\/em> bacteria. The observation of the inhibition zone and the\nformation of a clear zone around the disc evidence this. There are research results stated that the\nhigher the extract concentration, the greater the inhibition zone\nformed due to the greater content of antibacterial active compounds.18<\/sup>\nThe research is in line with the results of this study with the increasing\ninhibition zone value from the smallest to the largest concentration against S.mutans<\/em> and E.faecalis<\/em> (Table\n1).<\/p>\n\n\n\n

The\nresults of antibacterial research based on MBC in this study did not find MBC\nconcentrations in S.mutans<\/em> and E.faecalis<\/em> except in the positive\ncontrol group (chlorhexidine 0.2%). According to research by Nasri et al., a\ngood MBC value comes from the lowest extract concentration, which can reduce\nbacteria by 98%-99%.19<\/sup> The highest extract concentration in this\nstudy was at a concentration of 50%, reducing the number of bacterial colonies\n94.67% in S.mutans<\/em> and 84.19% in E.faecalis<\/em>. This shows that the 50% Batak\nonion bulb extract concentration can still not fulfill the MBC value requirements. Still, it does not rule out the\npossibility that at concentrations above 50% Batak onion bulb extract can kill S.mutans<\/em> and E.faecalis<\/em>. <\/p>\n\n\n\n

This\nstudy is in line with research regarding antibacterial extracts of Batak onion\nbulbs with 70% ethanol solvent at concentrations of 12.5%, 25%, 50%, and 70%\nagainst Escherichia coli<\/em>, which reported\nthat the highest inhibition zone diameter value was found in the 70%\nconcentration with a value of 8.70 mm. In comparison, the smallest inhibition\nzone diameter was found in the 12.5% concentration treatment with a value of\n6.53 mm.20<\/sup> This study is also consistent with earlier research on\nantibacterial extracts of Batak onion with 96% ethanol solvent at\nconcentrations of 100%, 50%, 25%, 12,5%, and 6,25% against Methicilin-Resistant\nStaphylococcus aureus<\/em> (MRSA) with inhibition of bacterial growth occurring\nat concentration\nlevels of 50% and 100% with inhibition zone diameters of 8,695 mm and 10,545 mm,\nrespectively. In contrast, extracts with 25%, 12,5%, and 6,25% concentration levels showed\nno inhibition.13<\/sup> In this study, an extract of Batak onion bulbs\nusing 70% ethanol solvent with different bacteria, namely S.mutans,<\/em> at a concentration of 0.78%, providing a better inhibition zone\nwith an inhibition zone diameter value of 9.00 mm. In comparison, the 0.78%\nextract concentration provided an inhibition zone of 8.06 mm against E.faecalis<\/em>.<\/p>\n\n\n\n

Research\non Batak onion bulb extracts against bacteria and different solvents has also\npreviously been carried out by Rubiatik et al., who reported that methanol extracts\nof Batak onion bulbs with concentrations of 0%, 1%, 5%, 10%, 15%, 20%, and 25%\nbegan to show inhibitory capability against Staphylococcus aureus<\/em> at a concentration of 1% with an inhibition\nzone diameter of 5 mm.21<\/sup> A research stated that a 96% ethanol extract from\nBatak onion bulbs with a concentration of 2.5% inhibited the growth of Escherichia coli<\/em>.2 <\/sup> Other research stated MRSA stopped growing when 96%\nethanol extract from Batak onion bulbs was added at a concentration of 50%.13<\/sup> T. Rhetso also stated that extract of Batak\nonion bulbs with Gas Chromatography-Mass Spectrometry (GC-MS) analysis at a\nconcentration of 1 \u03bcg\/ml could inhibit the growth of Staphylococcus aureus<\/em>,\nPseudomonas aeruginosa, and Aspergillus niger<\/em>.22<\/sup> All the research\nresults still show that the extract of Batak onion bulbs (Allium chinense<\/em> G.Don.) has antibacterial activity that can\ninhibit bacterial growth, even though the test bacteria and solvents differ. <\/p>\n\n\n\n

The\nresults of the phytochemical screening test in this study showed that a 70%\nethanol extract of Batak onion bulbs contained secondary metabolite compounds\nin the form of flavonoids, alkaloids, glycosides and saponins. Antibacterial\nand antibiofilm activity occurs due to the content of active compounds in the 70% ethanol extract of Batak onion\nbulbs that function as antibacterials. Flavonoids have a mechanism of action in\ninhibiting bacterial cell membrane function by forming complex compounds\nagainst extracellular proteins. Then, bacterial cell proteins will be\ndenatured, causing the cell membrane to be damaged and intracellular compounds\nto come out. Alkaloids can interfere with the components of the preparation of\nthe cell wall and bacterial DNA, causing the bacterial cell wall layer to not\nform intact and experience the cell’s death. Saponins can block the formation\nor transportation of each component to the cell wall and release cell contents,\nultimately inhibiting or killing bacterial cell growth.23-25<\/sup><\/p>\n\n\n\n

Based\non the Indonesian Pharmacopoeia VI edition, the diameter of the inhibition zone\nformed from an extract against bacteria is considered satisfactory if the\ndiameter produced is greater than 14 mm.26 <\/sup>In this study, the\ndiameter of the inhibition zone formed was greater than 14 mm against S.mutans<\/em> at concentrations of 25% (20.83\n\u00b1 1.36 mm) and 50% (24.66\u00b10.75 mm), while E.faecalis<\/em>\nbacteria were only found in the positive control (chlorhexidine 0.2%). However,\nif the results of the diameter of the inhibition zone are classified according\nto David and Stout, namely:23<\/sup> a). the category of no inhibition\nzone; b). weak inhibition is the zone of less than 5 mm; c). moderate is the\ninhibition zone of 5-10 mm; d). strong is the inhibition zone of 10-20 mm; e).\nvery strong, namely the inhibition zone of 20-30 mm, then the antibacterial\nextract of Batak onion bulbs against S.mutans<\/em>\nconcentrations of 0.78%, 1.56%, and 3.125% are in the category of\nmoderate inhibition, concentrations of 6.25% and 12.5% are in the category of\nstrong inhibition. Extracts with concentrations of 25% and 50% are in the\ncategory of very strong. On E.faecalis<\/em>\nobtained, extract concentrations of 0.78%, 1.56%, and 3.125% were included in\nthe moderate inhibition category, and concentrations of 6.25%, 12.5%, 25%, and\n50% were included in the strong inhibition category.<\/p>\n\n\n\n

In\nthis study, the extract of Batak onion bulbs with a concentration of 6.25% is\nthe optimal extract concentration because the concentration of 6.25% is the\nfirst extract concentration with a strong inhibition category and is the\nsmallest concentration significantly different in inhibition with a\nconcentration of 1.56% in the LSD post hoc test against E.faecalis<\/em> (p <0.05). In S.mutans<\/em>,\nit was also found that the concentration of 6.25% had a strong inhibition\ncategory and was the smallest concentration significantly different in\ninhibition against S.mutans<\/em> bacteria\nagainst the concentration of 3.125% (has moderate inhibition) based on the LSD\npost hoc test (p<0.05) (Table 5 and Table 6). <\/p>\n\n\n\n

The\naccumulation of biofilm occurs in two stages. The first stage is cell growth\nand forming Extracellular Polymeric Substances (EPS), so biofilm cells can accumulate.\nThe second stage is detachment or reattachment, using extracellular organs and\nproteins to sense and attach to the surface. Therefore, inhibiting or killing\ncells from increasing and preventing the formation of EPS is a way to prevent\nbiofilm growth through antibacterial substances contained in an extract.27<\/sup>\nThe presence of antibiofilm activity in this study is due to the 70% ethanol\nextract of Batak onion bulbs, which contains flavonoid, alkaloid, and saponin\nphytochemical compounds that work synergistically to damage biofilms and then\nkill planktonic bacteria that protect biofilms so that they can effectively inhibit\nbiofilm formation.28<\/sup><\/p>\n\n\n\n

The results of the antibiofilm test showed that 70% ethanol extract of Batak onion bulbs was able to inhibit biofilm formation at a concentration of 0.78% by 84.55% against S.mutans<\/em> and 84.71% against E.faecalis<\/em> and continued to increase until a concentration of 12.5%. The antibiofilm properties decreased at 25% and 50%, which is not in line with the greater the concentration, the higher the antibacterial effect. This can occur because the more chemical content in an extract (high concentration at a certain limit), the higher the antimicrobial effect, but if the content of compounds dissolved in the extract is too high (too high concentration), it can cause saturation, thus affecting the antimicrobial power of the extract. These results follow the hyperbolic curve theory, namely regarding the relationship between concentration and drug effects, which states that the higher the concentration of the drug, the higher the drug effect, but if it reaches the maximum effect at a certain concentration (optimal concentration), then the drug effect will decrease at a concentration higher than the optimal concentration.29<\/sup><\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

The MIC of 70% ethanol extract of Batak onion (Allium chinense<\/em> G.Don.) against the growth of S.Mutans and E.faecealis is at a concentration of 0,78% with an average inhibition zone diameter of  9.00\u00b10.43 mm and 8.06\u00b10.20 mm. The MBC and the Antibiofilm activity with 98-99% reduction in this study has not been found among the concentrations of the tested extracts. The highest concentration of 70% ethanol extract of Batak onion in this study is 50%, which killed and reduced the biofilm of S.Mutans at  94.67% and 83.99% and killed and reduced the biofilm of F.Faecalis<\/em> at 84.19% and 84.55%. Further research is required to prove that the concentration of 6.25%-50% of 70% ethanol extract of Batak onion is antibacterial against S.mutans<\/em> and 50% against E.faecalis<\/em>.<\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

None<\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

The author declares no conflict of interest.<\/p>\n\n\n\n

Funding Source<\/strong>s<\/p>\n\n\n\n

There are no funding sources.<\/p>\n\n\n\n

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