The Activities of Plant Extracts Heritage of Melayu Culture-Riau Archipelago on Bacteria Causing Diarrhea

Diarrhea is one of the major health problems in Indonesia. Lingga Malay ethnic in Riau archipelago use alternative medicines in a form of a obat pahit potion to treat diarrhea. This research aimed to determine the antibacterial activity of obat pahit extraction against Escherichia coli ATCC 11775, Salmonella ATCC 14028 and Shigella flexneri ATCC 12022. Antibacterial activity can be seen with the formation of inhibitory zone using diffusion method. The extract concentration used were 100%, 75%, 50% and 25%. Based on the antibacterial activity test, the largest inhibitory zone against E. coli was found at 100% Cnestis palala extract of 8.29 mm and the smallest inhibitory zone was found at 25% traditional medicine practioner (TMP) 2 SP4 of 5.73 mm. The more over, the largest inhibitory zone to S. was found at 100% Bauhinia semibifida of 8.81 mm; and the smallest inhibitory zone was found at 50% TMP 2 SP4 of 6.30 mm. Antibacterial activity with the largest inhibitory zone against S. flexneri at 75% C. palala of 6.21 mm and the smallest inhibitory zone was found at 50% TMP 2 SP4 of 5.62 mm.


Introduction
Diarrhea is one of the main health problems in developing countries including Indonesia. The disease can last for days and cause dehydration. Severe dehydration lead to weakness, shock and even death, especially in infants and children (Ganong, 1999). Diarrhea occurs due to several factors such as sanitation, nutrition, hygiene, malabsorption, chemical poisoning, allergies, viruses, parasites and bacterial infections (Suharyono, 2008 In an effort to deal with infection by microorganisms, it requires optimal working power of medicines with minor side effects. Current use of antibiotics is very high, but infection is still one of the global problems due to bacterial resistance to antibiotics. Thus, it is necessary to develop traditional medicines to support the improvement of public health (Ardiansyah, 2002).
The World Conservation Monitoring Center reports that the territory of Indonesia is an area with various types of medicinal plants that have been utilized to reach 2,518 species (Galingging and Bhermana, 2010). The knowledge of local people in the use of medicinal plants is different, namely in a simple and complex manner. In simple terms, the community uses one type of plant while in a complex way the community utilizes various medicinal plants or mixed ingredients called herbs to cure various disease complaints (Hanadari, 2014). This herb is made based on ancestral recipes, customs, beliefs, habits and traditional knowledge and experience (Dewoto, 2007

Materials and Methods
This research was carried out by making five extracts, namely three bitter medicinal herbs and two single herbs ((Kangkang Katup (Bauhinia semibifida) and Seven Lapis Root (Cnetis palala). Samples were obtained from Kalan Village, SP4 Village and Linau Village in Lingga District, Regency Lingga, Riau Islands and test bacteria were obtained from the UPT of the Health and Environment Laboratory of the Riau Province Health office.The five samples were made with a decoction extract with concentrations of 100%, 75%, 50% and 25%, as a negative control of 100 mL aquadest and positive control of chloramphenicol 30 µg The test bacteria suspension was prepared as much as 10 8 CFU/ml Test was carried out on MHA medium Repeated treatment was 3 replications and antibacterial activity of the extract was observed to inhibit the growth of diarrhea-causing bacteria through the inhibitory zone formed during incubation for 24 hours.

Plant Extracts Preparation
Bitter medicine and single herbs (simplicia) as much as 100 g in 1 L distilled water (infundation) in a container (cauldron) heated on a bath ± 15 minutes counted when the water began to boil, carried out for 3 consecutive days, after the first boiling (day 1), the sample is left in the container, to be boiled again on the second and third days (Nurhalimah et al. 2015) The results of the decoction were filtered using filter paper, 200 ml of extract was used and considered as 100% concentration, dilution was carried out to obtain concentration variations of 100%, 75%, 50% and 25% (Rahmawati and Siti 2014).

Nutrient Agar (NA) Medium
This media NA is used to breed bacteria, making it by weighing 20 g of NA dissolved in 1 L of distilled water using a stirring rod and heated on a hot plate. Sterilized using an autoclave with a pressure of 15 psi at 121 o C for 15 minutes (Aswarita 2013).

Mueller Hinton Agar (MHA) Medium
MHA media was used for testing antibacterial activity, making it with MHA weighing 38 g dissolved in 1 L distilled water using a stirring rod and heated on a hot plate. Sterilized using an autoclave with a pressure of 15 psi at 121 o C for 15 minutes.

Preparation of Test Bacterial Suspension
The test bacteria were inoculated as much as one ounce into 100 ml MHB from the inclined NA media and incubated 18-24 hours in a shaker incubator (150 rpm, 37 o C). The number of bacterial colonies grown in petri dishes was calculated using the total plate count (TPC) method to obtain 108 CFU/ml colonies (Sutton, 2011)

Antibacterial Activity Test (Disc Diffusion Method)
Test bacteria (108 CFU / ml) of 1 ml were inoculated into petri dishes, then poured 15 ml of MHA (pour plate) and allowed to solidify (Oktavia et al. 2013). Each extract was dripped on 100 µL sterile disc paper, allowed to stand for ± 15 minutes (Atikah 2013). After condensing media positioned on the surface of the media: herb extracts and medicinal plants with various concentrations (100%, 75%, 50% and 25%), sterile distilled water as much as 100 µL (negative control) and chloramphenicol 30 µg (positive control). After being incubated for 24 hours, antibacterial activity was observed by measuring the diameter of the inhibitory zone (mm) using a calipse.  (2016), the diameter of the inhibitory zone was <5 mm, the activity was categorized as weak, the diameter of the inhibition zone was 6-10 mm categorized as moderate, the diameter of the inhibition zone was 11-20 mm categorized as strong and the diameter of the inhibitory zone> 21 mm was categorized as very strong.

Result and Discussion
In this study to obtain extracts, the infundation method is used, namely the method of extraction of heat using water solvents. In Table 1 Rahmawati and Siti (2014), that the higher the concentration, the greater the antibacterial activity.On the herb extract of POT 1 Kalan, the largest inhibition zone was obtained from the concentration of 75% (6.58 mm) on the other hand at a concentration of 100% inhibitory zone, namely 6 Based on the antibacterial activity, it was found that extract activity was not directly proportional to the level of concentration. It was thought that the extract did not work stably in inhibiting the growth of S. typhi, indicated by the unequal activity between levels of concentration that can be seen from the inhibition zone formed. Negative control and positive control were used as a comparison in determining antibacterial activity from extracts of bitter and single herbs. Negative control was used by distilled water to see the effect of solvent on the extraction stage on the resulting inhibition zone. Positive control uses chloramphenicol, a broad-spectrum antibiotic. According to Pelczar & Chan (2008) chloramphenicol is relatively non-toxic when used in mammals therapeutically.

Conclussion
Based on the research conducted, the following conclusions were obtained: 1. The biggest inhibitory zone against E. coli ATCC 11775 was obtained from C. palala extract which was 8.29 mm at a concentration of 100% and the smallest inhibition zone of 5.73 mm from POT 2 SP4 at a concentration 25%.
2. The biggest inhibitory zone against S. typhi ATCC 14028 was obtained from B. semibifida extract which was 8.81 mm at a concentration of 100% and the smallest inhibition zone of 6.11 mm from POT 3 Linau at a concentration of 25%.