IN VITRO PHARMACOLOGICAL ACTIVITY TEST OF TELANG FLOWER KOMBUCHA AS ANTIBACTERIAL Vibrio cholerae AND Shigella dysenteriae THROUGH FERMENTATION

Diarrhea that occurs in the digestive tract is a big problem for human intestinal health. This can be caused by the activity of pathogenic bacteria in the form of Vibrio cholera and Shigella dysinteriae . In this study, kombucha has been shown to have pharmacological activity in vitro in inhibiting the growth of Vibrio cholera and Shigella dysinteriae bacteria at all sugar concentrations. The purpose of this study was to determine the in vitro pharmacological activity of the butterfly pea flower kombucha in inhibiting the growth of the two test bacteria consisting of a sugar concentration of 20%, 30% and 40%. The method for testing the inhibition of both bacterial growth tests is disc diffusion. One way ANOVA and post hoc analysis are one of the quantitative test methods used in this study of the average diameter of the inhibition zone of teal flower kombucha from various sugar concentrations. The results of this study have proven based on a one-way ANOVA with each P value> 0.05 and then continued through post hoc analysis concluded that the telang flower kombucha at a concentration of 40% is the best treatment in inhibiting the growth of both test bacteria where the concentration is significantly different from sugar concentration 20% and 30%.

Telang flower kombucha, as has been described in the results of previous studies, has provided a lot of health effects that are quite good in fighting diseases and infections caused by the activity of pathogenic bacteria [15]  fermentation is to cover the glass jar using a cloth cover so that the kombucha fermentation process runs statically for 2 weeks at room temperature conditions [28].

Data Analysis
The

Shigella dysenteriae
The Shapiro-Wilk test 0,66 The fourth step in the butterfly pea flower kombucha fermentation is adding the sugar concentration according to the treatment which includes 20%, 30%, and 40%.The fifth step in the butterfly pea flower kombucha fermentation is to heat the sugar until it boils within 10 minutes and then put it in a glass jar for each sugar concentration treatment.The sixth step in the butterfly pea flower kombucha fermentation is to put the cooking water on the inside of the glass jar where sugar has been added based on the concentration of each.The seventh step in the fermentation of Firman Rezaldi, et.al.134 butterfly pea flower kombucha is to cool the cooking water at 25 0 C then add the initial kombucha culture with 1 week of age as much as 8% (v/v) in each treatment.The eighth step in the butterfly pea flower kombucha steps in testing the inhibition of Vibrio cholera and Shigella dysintreae growth were preparing 24 petri dishes to be poured into Muller Hinton Agar (MHA) media in each 15 mL petri dish.Silence the media until it solidifies.Dipping a sterile cotton swab into the suspension of Vibrio cholerae and Shigella dysenteriae bacteria.Wipe the MHA media until it is completely covered on the surface.Attaching the disk that has been soaked to the preparation in the form of a solution of fermented eggplant kombucha at various concentrations, namely the first petri dish containing the eggplant kombucha at a sugar concentration of 20%.The second petri dish contains tethera kombucha at a sugar concentration of 30%.The third petri dish contains tethera flower kombucha at a sugar concentration of 40%.The fourth petri dish contains kombucha fermented black tea as a positive control and the fifth petri dish contains sterile distilled water as a negative control.Each treatment was repeated 3 times.Incubate the media containing the antibacterial agent and the test bacteria for 1 day.Measuring the average diameter of the inhibition zone from each concentration of sugar in telang

Butterfly pea flower kombucha used as an
active substance in this study has been shown to have pharmacological activity in vitro to inhibit the growth of both test bacteria.This is because telang flower kombucha contains several secondary metabolites consisting of alkaloids, flavonoids, and saponins which have the potential to inhibit pathogenic bacteria both from gram-positive and gram-negative bacteria [36].The flavonoid group contained in kombucha butterfly pea flowers has a mechanism of action as an antibacterial including inhibiting nucleic acid synthesis, cell membrane function, energy metabolism which can cause permeability damage to the bacterial cell wall, microsomes, lysosomes which are the result of interactions with bacterial plasmids.The formation of complex compounds in extracellular proteins capable of bacterial cell membranes is one of the working mechanisms of flavonoids as an antibacterial source in inhibiting the function of cell membranes of pathogenic bacteria [36].The alkaloid group contained in telang flower kombucha has a mechanism of action as an antibacterial source, including by interfering with the peptidoglycan component in bacterial cells, so that the cell wall layer is not completely formed and causes lysis and even cell death in pathogenic bacteria.The class of secondary metabolite compounds in the form of saponins contained in telang flower kombucha as an antibacterial source, among others, is by causing a leak in the protein and enzyme parts found in the cell walls of pathogenic bacteria.Saponins are also able to reduce surface tension which can lead to increased permeability and cell leakage as a result of released intracellular compounds [37].Saponins have the potential to diffuse through the outer membrane and cell wall which are prone to bind to the cytoplasmic membrane until disturbance or decrease in the stability of the cell membrane of pathogenic bacteria occurs.This causes the cytoplasm to leak, resulting in cell death for the pathogenic bacteria themselves [31].CONCLUSION From this study it can be concluded that the butterfly pea flower kombucha has been shown to have pharmacological activity in vitro in inhibiting the growth of both test bacteria and a sugar concentration of 40% in the butterfly pea flower kombucha is the best treatment in inhibiting the growth of both test bacteria compared to the butterfly pea flower kombucha at the sugar concentration.by 20% and 30% as well as a positive control.

Preparation of Butterfly Pea (Clitoria ternatea L.)
butterfly pea flower kombucha fermentation is to weigh 7.2% of the water until 2.4% of the water remains.

Table 1 .
Mean Measurement Results of the Inhibition Zone Diameter of Muller Hinton Agar (MHA) Media

Table 2 .
Data Normality Test Through the Shapiro Wilk Test