INTERNASIONAL CONFERENCE ON
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php/icmahs Antibacterial Activity of Rosella Tea Kombucha Against The Growth of Escherichia coli Based on Fermentation Time Variations Putu Diah Gayatri Purnama Dewi1.
Gusti Ayu Made Ratih2*.
Putu Ayu Suryaningsih3 1,2,3 Department of Medical Laboratory Technology.
Health Polytechnic of Denpasar *Corresponding author: iga_ratihkurada@yahoo.
Article history Posted :
Reviewed :
Received :
2023-11-03
2023-11-02
2023-10-24
ABSTRACT
Background: Public awareness has begun to experience changes in maintaining consumption patterns, such as consuming functional drinks.
Rosella Tea Kombucha is able to produce important compounds for the body such as polyphenols, organic acids, vitamin B complex, vitamin C, enzymes and antibiotics.
The use of antibiotics as a treatment for Escherichia coli can cause side effects and resistance so this Rosella Tea Kombucha can be an The aim of this research was to determine the activity of Rosella Tea Kombucha as an antibacterial against Escherichia coli bacteria.
Methods: This type of research is a true experimental with a posttest-only group Antibacterial test uses the disc diffusion method with the Ciprofloxacin antibiotic as positive control and sterile distilled water as negative control.
Results: The results showed that Rosella Tea Kombucha was able to inhibit the growth of Escherichia coli bacteria based on variations in fermentation time on 6, 8, 10, 12 and 14 days with the diameter of the inhibition zone respectively being 7.
27 mm.
73 mm.
Statistical tests using the One Way ANOVA test obtained a sig of .
< 0.
05, which means there are significant differences in the growth inhibition zone of Escherichia coli at various fermentation times of Rosella Tea Kombucha.
The largest inhibitory zone diameter was shown in the fermentation time variation of 14 days with strong inhibitory category.
Keywords: Antimicrobial Activity.
Rosella Tea Kombucha.
Fermentation Time Variations.
Putu Diah Gayatri Purnama Dewi, et all : Antibacterial Activity of Rosella Tea Kombucha Against The Growth of Escherichia coli Based on Fermentation Time Variations INTERNASIONAL CONFERENCE ON MULTIDISCIPLINARY APPROACHES IN HEALTH SCIENCE VOLUME 1 TAHUN 2023.
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php/icmahs INTRODUCTION Kombucha tea is a traditional drink that has unique characteristics, because the process to make this tea by fermenting natural ingredients or various types of tea assisted by symbiotic culture.
The symbiotic culture used has various names, one of which is Symbiotic Culture of Bacteria and Yeast (SCOBY).
The kombucha culture or SCOBY is round and pale white and has a rubbery, gel-like texture.
This follicle culture is made from cellulose resulting from the metabolism of acetic acid bacteria which is able to float and sometimes sink in the kombucha liquid.
In general, kombucha is known as tea mushroom which is fermented for 1-14 days or according to the desired acidity level.
the fermentation of kombucha tea, the growth of yeast and bacteria occurs, the sugar content increases, ethanol production increases, and organic acid production increases (Restuati.
The results of fermentation and oxidation of microorganisms in kombucha are useful for preventing cancer, improving liver function, helping control high blood pressure, preventing stroke, relieving throat pain, reducing fat and cholesterol, maintaining body stamina and maintaining skin cleanliness.
The antimicrobial content in kombucha can inhibit the growth of Shigella sonnei.
Escherichia coli, and Salmonella The combination of glucuronic acid and lactic acid in kombucha is very effective at destroying microorganisms .
acteria, viruses and fung.
Apart from that, the acetic acid contained in kombucha is known to be able to inhibit and kill a number of gram-positive and gram-negative bacteria (Naland, 2.
Kombucha tea contains various compounds that are very important for the body, such as polyphenols, organic acids .
cetic acid, gluconic acid, lactic aci.
, vitamin B complex, vitamin C, enzymes and antibiotics (Firdaus et al.
, 2.
With the various ingredients contained in kombucha, it is believed that kombucha can overcome health problems such as high blood pressure, rheumatism, obesity, arthritis, migraines, diabetes and others (Naland, 2.
Kombucha tea can be mixed and matched with various kinds of tea, for example Rosella is an annual plant with single leaves, oval-shaped and has serrations on the edge of the leaves.
In general, the length of Rosella leaves ranges from 6-15 cm and width 58 cm.
Rosella has a single flower where on each flower stalk there is one flower bud which has 811 hairy petals (Haidar, 2.
Rosella as a health drink is believed to cure various diseases such as hypertension, diabetes, diuretic.
Rosella flower petals contain active substances such as hibisci Anthocyanins are compounds that can produce a red color and are classified as flavonoids.
Anthocyanins are believed to be able to cure degenerative diseases (Djaeni et al.
, 2.
Apart from that, rosella contains catechins, vitamins C.
B1.
B2, carotenoids, organic acids, saponins and alkaloids which can damage bacterial cell proteins and cause cell death.
With this antibacterial content, it is able to kill several types of bacteria such as Pseudomonas Klebsiella Staphylococcus aureus and Escherichia coli (Arifianti, 2.
Escherichia coli is a bacteria belonging to the Enterobacteriaceae.
Escherichia coli bacteria are normal flora in the human body which can be pathogenic, causing various diseases such as Putu Diah Gayatri Purnama Dewi, et all : Antibacterial Activity of Rosella Tea Kombucha Against The Growth of Escherichia coli Based on Fermentation Time Variations INTERNASIONAL CONFERENCE ON MULTIDISCIPLINARY APPROACHES IN HEALTH SCIENCE VOLUME 1 TAHUN 2023.
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php/icmahs Escherichia coli bacteria is the cause of diarrhea with a percentage of 21.
5% apart from Shigella dysenteriae.
Salmonella spp.
Vibrio Cholerae and Campylobacter jejuni (Estri, 2.
In 2020, the coverage of diarrhea sufferers throughout Indonesia reached 44.
4% at all ages and 28, 9% in toddlers (Kementerian Kesehatan RI , 2.
The use of synthetic antibiotics as a treatment for diseases caused by Escherichia coli bacteria can cause side effects and resistance.
Alternative treatment is needed as an antibacterial using traditional medicine made from natural ingredients that are safer for daily consumption, namely Rosella Tea Kombucha.
Therefore, in this study, the antibacterial activity of Rosella Tea Kombucha was tested against Escherichia coli bacteria using the disc disk test method based on varying fermentation times of 6, 8, 10, 12 and 14 days.
METHOD
This research is true experimental research with the design used is Posttest-Only Group Design, namely by measuring the inhibition zone of five types of Rosella Tea Kombucha samples with varying fermentation The research was conducted at the Bacteriology Laboratory.
Department of Medical Laboratory Technology.
Health Polytechnic of Denpasar from February to June 2022.
The sample used in this research was Rosella which was made into kombucha.
The Rosella used is 250 grams of dried Rosella flowers obtained from Denpasar area where the parts taken are the flower petals to be made into tea.
For kombucha, we use kombucha seeds from Inidhya Kombucha in the Sesetan area.
South Denpasar.
This research is an experimental research in which the treatment used is the fermentation of Rosella Tea Kombucha with the same concentration but different fermentation times, namely samples on the 6th, 8th, 10th, 12th, 14th day, positive control .
ntibiotic ciprofloxaci.
and negative control .
terile distilled wate.
This test was 3 replication at each time variation with the same concentration, 15 treatment sample data were Positive and negative controls were repeated 2 times to obtain 4 control data.
The total data obtained was 19 data.
The instruments used in this research were a digital balance, analytical balance (RADWAG AS220.
, cover glass, stir bar, gas stove, beaker (IWAKI TE-.
, disposable Petri disk (ONEMED), vernier caliper, hot plate (JISICO), magnetic stirrer, measuring pipette (IWAKI), ball pipette, test tube (IWAKI), tube rack, tube, autoclave (TOMY SX-.
Erlenmeyer (IWAKI), dropper pipette, incubator (ESCO Isother.
, spirit lamp .
Mc Farland Densitometer .
iosan DEN-1B).
Biosafety Cabinet (BSC-1800 II B2-X).
The materials used in this research were dried rosella.
SCOBY, sugar, mineral water, blank disk (OXOID).
Escherichia coli bacteria, kombucha starter solution, distilled water.
MHA media (OXOID).
Ciprofloxacin disk (OXOID), physiological NaCl solution 0, 85%, 70% alcohol, sterile cotton cap, aluminum foil.
Mc Farland standard 0.
Procedure for making Rosella Tea Kombucha Boil 250 grams of dried Rosella in 2500 ml of water until it turns red.
10% sugar from the total volume is added to the roselle tea solution then stirred until homogeneous then waited until the solution cools .
emperature below 50oC).
The cooled Rosella tea is divided into 5 different containers, amounting to 500ml per Putu Diah Gayatri Purnama Dewi, et all : Antibacterial Activity of Rosella Tea Kombucha Against The Growth of Escherichia coli Based on Fermentation Time Variations INTERNASIONAL CONFERENCE ON MULTIDISCIPLINARY APPROACHES IN HEALTH SCIENCE VOLUME 1 TAHUN 2023.
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php/icmahs container, then added SCOBY and 10% starter of the total volume per container, then covered with gauze.
Each container is labeled according to the fermentation day, namely days 6, 8, 10, 12 Kombucha tea that has been labeled is fermented according to the specified time and taken according to the time period stated on the Antimicriobial test against Escherichia coli Empty discs were saturated with kombucha tea according to the day of fermentation, then placed on an empty plate.
The sterile cotton swab that has been prepared is then dipped into the prepared bacterial suspension, then lifted and squeezed by attaching the cotton swab to the wall of the Bacteria are inoculated on the surface of the MHA media until they are evenly distributed over the entire surface of the media and the media is closed again.
The media that has been inoculated with the suspension is left for 15 minutes so that the bacterial suspension can seep into the media.
Discs that had been saturated from each kombucha tea according to the day of fermentation were attached to the inoculated MHA medium using tweezers.
Discs that have been attached to the media surface must not be moved or shifted.
Positive controls and negative controls were attached to MHA media that had been inoculated which was different from the kombucha tea media.
The media to which the disc was attached was incubated at 37AC for 24 hours in an inverted Data was collected through laboratory examination by measuring the diameter of the inhibition zone using a caliper.
Measurements were carried out on the growth of Escherichia coli bacteria formed from various samples, namely fermentation results on the 6th, 8th, 10th, 12th and 14th days expressed in millimeters .
The data analysis used in this research is quantitative analysis, carried out using the One Way ANOVA statistical test to determine the differences between various variations in the fermentation time of Rosella Tea Kombucha in the growth inhibition zone of Escherichia coli.
To determine the significant differences in the inhibition zone between each of the variations in the fermentation time of Rosella Tea Kombucha which can inhibit the growth of Escherichia coli, the Least Significant Deference (LSD) test was used.
RESULTS
Results of the Rosella Tea Kombucha inhibition zone test on the growth of Escherichia coli bacteria with five treatments, namely fermentation days 6, 8, 10, 12 and 14 with 3 replication using the disc diffusion method.
The results showed inhibition of the growth of Escherichia characterized by the formation of a clear zone around the disc that had been given the test substance, namely Rosella Tea Kombucha on MHA media.
Table 1.
Results of Antibacterial Activity of Rosella Tea Kombucha Fermentation Time Inhibition zone.
(Day.
Mean of Inhibition Zone Inhibition Zone Category .
i 7,60 7,70 6,50 7,27 Medium
7,90
8,40
7,60
7,97
Medium
9,00
8,50
7,80
8,43
Medium
9,50
9,90
9,30
9,57
Medium
11,60
13,10
13,50
12,73
Strong Putu Diah Gayatri Purnama Dewi, et all : Antibacterial Activity of Rosella Tea Kombucha Against The Growth of Escherichia coli Based on Fermentation Time Variations INTERNASIONAL CONFERENCE ON MULTIDISCIPLINARY APPROACHES IN HEALTH SCIENCE VOLUME 1 TAHUN 2023.
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php/icmahs Based on table 1, there are differences in diameter results from the first to the third The diameter category of Rosella Tea Kombucha's inhibitory zone on the growth of Escherichia coli bacteria was the largest produced on the 14th day of fermentation .
50 m.
with strong category, while the smallest diameter was produced on the 6th day of fermentation .
50 m.
with medium Meanwhile, the results obtained from control group .
95 m.
in the sensitive category and in the negative group there was no inhibition zone produced.
6 Days 8 Days 10 Days 12 Days LSD
ANOVA
Fermentation Time Significant Time (Day.
(Day.
Sig.
In this study, a normality test was carried out using the Shapiro-Wilk test.
In this test the data is said to be normally distributed .
because all the data gets a Sig > 0.
Based on the results of the One Way ANOVA test, a sig of .
< 0.
05 was obtained, which means that there were differences in the growth inhibition zones of Escherichia coli at various times of rosella tea kombucha fermentation.
Furthermore, the LSD results from the five fermentation time variations showed that there were significant differences in the inhibition zone on the 14th day of fermentation compared to the other time Picture 1.
Rosella Tea Kombucha Table 2.
One Way ANOVA and LSD Statistical Test Results 14 Days Inhibition Zone .
Picture 2.
Antibacterial Activity Graph of Rosella
Tea Kombucha
DISCUSSION