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Research Title: “Feasibilty of Saluyot Corchorus olitorius Leaves Extract as Antibacterial Dishwashing Liquid”
Student Researchers:
Katherine Jane A. Siarot (Team leader)
Luchie R. Mascariola
Sienna M. Lantaca
INTRODUCTION
People living in developing countries depends primarily on traditional plants as a source for treating diseases by killing microorganisms such as E. coli and Salmonella that causes diseases and illness in the body. One of such common plant available in developing countries like Philippines is Corchorus olitorius also known as Jute or Saluyot. C. olitorius is edible and has a mucilaginous texture. Its leaves are rich in betacarotene, iron, calcium, and vitamin C. It has an antioxidant activity with a significant ?-tocopherol equivalent vitamin E. With the use of this traditional plant, the researchers would like to test the effectiveness of Saluyot leaves extract in killing such microorganisms as mention above.

Escherichia coli are a Gram-negative, facultative anaerobic, rod-shaped, coliform bacterium of the genus Escherichia found in the lower intestine of warm-blooded organisms (endotherms). Most of the strains are harmless, but some can cause serious food poisoning. It constitutes about 0.9% of gut microbiota, and fecal-oral transmission is the major route of this bacterium cause disease. This can be grown and/or cultured easily and inexpensively in a laboratory. Salmonella is another microorganism from a genus of rod-shaped (bacillus) Gram-negative bacteria of the family Enterobacteriaceae. Species are non-spore-forming, predominantly motile enterobacteria with cell diameters of about 0.7 to 1.5 µm, and a length of 2 µm to 5µm, and peritrichous flagella. Also a facultative anaerobes, capable of generating ATP with and without oxygen. Species are intracellular pathogens and certain serotypes cause illness. It can transfer from animal-to-human and from human-to-human; and usually invade only in the gastrointestinal tract and can cause food poisoning, typhoid fever, and paratyphoid fever.

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Nowadays, washing dishes is one of the major chores that people are facing every day. The real purpose of this study is to produce a product out of Saluyot leaves extract as an antibacterial agent in an antibacterial dishwashing liquid. With these, the student researchers will be evaluating the Saluyot leaves extract as a potential antibacterial agent. This study shows how simple organic materials that are easily found elsewhere can turn into something useful as antibacterial dishwashing liquid.
MATERIALS AND METHODS
Experimental Design
In the experimental set-up, six petri dishes were used and divided into two: three petri dishes for E. coli; and three petri dishes for Salmonella. Each petri dish is divided into four quadrants; each quadrant is mark with the corresponding experimental plant concentrations and the control group. Every quadrant contains filter paper soaked in the Saluyot leaves extract.

Preparation Plant Sample
The Saluyot (C. olitorius) leaves were collected from Purok-3, Antequera, Nabunturan Compostela Valley Province. The collected leaves of (C. olitorius) were washed with running water and were air-dried for two days before homogenizing it using a blender.

Preparation of the C. olitorius Leaf Ethanolic Extract
The dried leaves were homogenized or powderized; weight was measured. The homogenized leaves were soaked in ethanol for 36 hours in a tightly closed container with a ratio of 1:2 or 500 mL of homogenized leaves and 1000 mL ethanol. After 36 hours, the soaked leaves were filtered and the filtered extract were stored in a clean container with cover.

Modified Evaporation of Ethanolic Extract
The modified evaporation process was performed after extract were being filtered; this process will separate the ethanol from the extract. The extract of C. olitorius were heated in a hot plate at 380°C for 4 hours. After this process, the filtrate extract were store in the refrigerator until further use.

Collection of Microorganism
The microorganisms E. coli and Salmonella were purchased from the Department of Science and Technology (DOST), Davao City, Philippines. The purchased microorganisms were stored in the Biotechnology Laboratory of the school campus (NNCHS) until its further use.

Preparation of the Nutrient Agar
In a beaker, pour 25g of agar in a 1L of distilled water. Mix the solution through stirring while being heated. The nutrient agar was distributed into different petri dishes equally and was set to cool down.

Sterilization of Laboratory Materials
The laboratory materials were sterilized before and after experimentation to avoid contamination of bacteria in the working place. The materials were placed in the autoclavable bag and were sterilized in the autoclave at 121°C at 15 psi.

Inoculation of the Bacteria
Each petri dish with the nutrient agar were divided into four quadrants labelled as 35%, 75%, 50%, and control group. The bacteria E. coli and Salmonella were placed evenly in the petri dish using an inoculating loop. The Saluyot leaves extracts were also divided into different concentrations 35%, 75%, and 50% respectively. Filter papers with 6mm in diameter were soaked in the different concentrations of Saluyot leaves extracts and were placed in the centre of each quadrant. The experimental set-up were placed in the laminar flow hood for about 24 hours, and result were observed and were recorded.

Procedures in Making Dishwashing Liquid
The raw materials in making the dishwashing liquid are distilled water, MC – Gel, bubble enhancer, degreaser, industrial salt, scent, colorant, preservative, and antibacterial agent. The following raw materials mentioned except from antibacterial agent were purchased from the nearest local market that has availability of these materials. The antibacterial agent is the observed effective concentration of Saluyot leaves extracts as antibacterial against E. coli and Salmonella.

The formulation in making the dishwashing liquid are 15 litres of distilled water, 1.8 kg. of MC – Gel, 45 mL of colorant and scent, 375 g of industrial salt (divide into 2 halves), 600 mL of bubble enhancer (CD – Amide), 150 mL of Degreaser (BC – 06), 19.5 mL of antibacterial agent and preservative.

In container, put a little amount of water and dissolve the other half of the salt, and mix well. In another a plastic container, mix MC – Gel and one – half of the salt completely, and add water slowly while stirring the mixture. Add a desired colorant, degreaser, and bubble enhancer one at a time, and then mix the solution thoroughly. Next, add the desired scent, antibacterial agent, and preservative, and add the salt that is dissolved in water. Set aside for 24 – 36 hours before storing it in airtight bottle.

https://www.slideshare.net/mobile/jaycastillo/how-to-make-dishwashing-liquid-29700489
Decontamination Process
The materials used during the experiment were decontaminated, were wrapped, and were placed in the autoclavable bag. The bag were placed inside the autoclave for 15 minutes and the materials were washed thoroughly after.
Data Analysis
The statistical tool that were used in the study is the Analysis of Variance (ANOVA) to determine the significant difference of the mortality of bacteria in the experimental groups in comparison with the control groups. The Mean were also used in determining the average zone of inhibition of E. coli and Salmonella subjected to each concentrations of the extracts.

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References
1Y. Evale, M.A.C. Marmito, L.D. Perlas (January 2018). THE EFFICACY OF SANTOL
(Sandoricum koetjape) LEAVES EXTRACT AS AN ANTISEPTIC GEL AGAINST
Staphylococcus aureus AND Staphylococcus epidermis.

2H.D. Kusumaningrum, M.M. van Putten, F.M. Rombouts, and R.R. Beumer (2002).

EFFECTS OF ANTIBACTERIAL DISHWASHING LIQUID ON FOODBORNE
PATHOGENS AND COMPETITIVE MICROORGANISMS IN KITCHEN
SPONGES. Retrieved from:
https://www.ncbi.nlm.nih.gov/m/pubmed/11811157/3HOW TO MAKE DISHWASHING LIQUID. Retrieved from:
https://www.slideshare.net/mobile/jaycastillo/how-to-make-dishwashing-liquid-29700489
4www.healthycleaning101.org/information_about_soaps_and_detergents/5 www.health.state.mn.us/handhygiene/how/bestsoap.html6 https://medlineplus.gov/ecoliinfections.html7https://www.foodsafety.gov/poisoning/causes/bacteriaviruses/ecoli/8https://www.niaid.nih.gov/diseases-conditions/e-coli9https://www.mayoclinic.org/diseases-conditions/e-coli/symptoms-causes/syc-2037205810https://www.cdc.gov/ecoli/index.html11https://ecdc.europa.eu/en/escherichia-coli-ecoli/facts12K. Sen, A. Goel, S. Rawal, N. Mahajan, S. Baboo, S. Alok, (January 2010).

ANTIMICROBIAL ACTIVITY OF BASELLA RUBRA LEAVES. Retrieved from:
https://www.researchgate.net/publication/238722045_ANTIMICROBIAL_ACTIVITY_OF-BASELLA_RUBRA_LEAVES

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