Singapore Piaget Academy No. 20 Jalan Jlopo Solo Baru Indonesia

Rodenticide from Pong Pong (Cerbera odollam) Fruits

Cherry , Jonathan , Albert & Alfian

Introduction Rat infestation pose a significant health risk to humans and other animals. In In-

donesia, the population of rats has been increasing and this has caused the spread-ing of diseases like Salmonella, Weil’s disease, E.coli and TB in some areas of the country. They also carry fleas, mites and ticks and cause acute allergic reactions.

The damage to rice plants by rats is by far the greatest agricultural problem in the country. The ricefield rat was ranked as the most important non-weed pest between 1986 and 2000, followed by the rice stem borer, rice leaf folder and brown planthop-per (Source: Directorate of Food Crop Protection, Government of Indonesia). A re-cent conservative estimate is that typically between 5 and 15% of the world rice crop is lost because of rodents. In Southeast Asia, the reported impact of rodents is highest in Indonesia, where Geddes (1992) attributed approximately 17% of rice production as lost to the ricefield rat

With these, the researchers have thought of using Pong-Pong fruits (Cerbera odollam) as potent rodenticide to kill or ward off rats at home and some agricultural areas in the country. If this type of plant which is endemic in India and certain parts of Southeast Asia will be proven potent, it will provide a very practical and cost-effective rodenticide for all. Statement of the Problem

This study primarily aims to develop rodenticide from Pong Pong fruits and seeds.

Specifically, this seeks to answer the following questions. 1.What is the active constituent present in Pong Pong fruits? 2.What is the level of toxicity of Cerbera odollam seeds? 3.What is the LD50 (lethal dose) of the rodenticide from Pong Pong ? 4.What are the effects of Pong Pong rodenticide on rats? 5.Is there significant difference between rodenticide from Pong Pong and commer-

cial rodenticides?

Hypothesis Ho = There is no significant difference between rodenticides from pong pong and

commercial rodenticides. Ha = There is significant difference between rodenticides from pong pong

Methodology A. Gathering of Materials The researchers gathered the Pong-Pong fruits from the school vicinity and labor-atory apparatuses and equipment were taken from the school laboratory. B. Preparation of Pong Pong Rodenticide The researchers separated the skin and seeds of the Pong Pong fruits and 3 kil-ograms of the plant material were obtained. The plant material was placed in a blender and 3 liters of 1:1 ratio of methanol/water were used to facilitate the meth-anol extraction. The extracted juice from Pong Pong was separated from its residue and was dried under the sun for several hours to evaporate the solvent. After few hours the solution turned black. The Pong Pong extract was further subjected to water bath for complete evaporation of methanol and to recover the crude extract containing the active components. It was then mixed with leftover rice and fish, made into little pellets and sun-dried before it was administered to laboratory mice. C. Testing the Pong Pong and Commercial Rodenticides to Mice Five grams of pong pong rodenticides were administered daily to each rat in the experimental group and another five grams of com-mercial rodenticides were administered to each rat in the control group. The mice in the experimental group and control group were provided water daily. Symptoms were observed and recorded from each rat in the experimental and control group based on body weaknesses, vomiting, diarrhea, and other symptoms.

Results and Discussion A. Active Constituent of Ponpong The kernels of C. odollam contain cerberin, a digoxin-type cardenolide and car-diac glycoside toxin that blocks the calcium ion channels in heart muscle, causing disruption of the heart beat, most often fatal. The difficulty in detecting cerberin in autopsies and its ability of strong spices to mask its taste makes it an agent of homi-cide and suicide in India: in 2004, a team led by scientist Yvan Gaillard documented more than 500 cases of fatal Cerbera poisoning between 1989 and 1999 in the southwest Indian state of Kerala alone.

B. The LD50 Analysis of Rodenticide from Pong Pong Based on the results gathered from the experimental and control group, the researchers found out that there was no siginificant difference between the rodenticide from pong pong made up of cerberin glycoxide toxin and commercial rodenticide with permethrin. The LD50 (lethal dose) of the rodenticide from the experimental and control groups is 323 mg/kg. This is the amount of the active ingredients from both rodenticides that is enough to kill 50% of the test specimens C. Test on the Effectiveness of Pong Pong Rodenticide Against Commercial

Rodenticide on Rats

The table above shows the test of effectiveness of Pong Pong rodenticide against commercial rodenticide. The specimen 1 in the experimental group took 10 days to see the toxic effect of the rodenticide form pong pong while the specimen in the control group which received the commercial rodenticide took 7 days. On the other hand , the specimen 2 in the experimental group took 6 days while in the control group took 5 days. Finally, the last specimen in the experimental group took 7 days while the specimen in the control group took 6 days before they die after the administration of pong pong and commercial rodenticides respectively. The mean days for the Experimental group was 7.67 while the Control group was 6. The standard deviation for the experimental group was 1.7 and the control group’s standard deviation was 0.82. There was a difference in the mean of 1.67 and a difference of 0.88 in the standard deviation. Further analysis revealed a computed t value of 1.53 which is less than the tabular value of 2.776 at degrees of freedom 4 and 0.05 significance level. Since the computed t value is less than the tabular value, the researchers accepted the null hypothesis which states that there is no significant difference between Pong Pong rodenticide made of cerberin and commercial rodenticide made of permethrin. Summary, Conclusion and Recommendation The researchers prepared the pong pong fruits by peeling and separating the skin and the seed. The obtained sample was placed in the blender and methanol was added to facilitate the extraction procedure. The extract was brought to water bath and obtained the pong pong crude extract. The crude extract from pong pong and the commercial rodenticide were added with left over rice and shredded fish and was administred equally to the laboratory rats in the experimental and control group. Prior to the feeding of the rats, the researchers made sure that the specimens are of similar in age, gender and weight and each cage had an ample supply of potable water. Based on the results, it was proven that the pong pong rodenticide administered to laboratory rats took an average of 7 days. The commercial rodenticides on the other hand took 6 days for the rats to die after the administration of the rodenticide. This proves that the effect and level of effectiveness of the rodenticides from Pong Pong and commercial rodenticides was the same as it only took a mean difference of 1 day. Therefore, people should propagate this plant as its cerberin content is comparable to expensive commercial rodenticide. .

Test Specimen Experimental Group (Using Experimental Group

T1 10 days 10 days

T2 6 days 6 days

T3 7 days 7 days

Mean X1=7.67 X1=7.67

t-test = 1.53 tabular value= 2.766 df= 4

Introduction

PONG PONG, also known scientifically as Cerbera odollam, is a plant species in the Family Apocynaceae. It yields a potent poison which has been used for suicide and mur-der. Using this plant, we want to create an antibiotic which can treat infections and ill-nesses caused by different types of bacteria. Such illnesses as syphilis, tuberculosis, sal-monella, and some forms of meningitis are caused by bacteria.

Antibiotics usually are made from bacteria, fungi and a bit of chemistry. However, the method is quite expensive which can lead to high price of medicine. It is for these reasons that the researchers are trying to find a cheaper substitute for commercial anti-biotics in the market.

Statement of the Problem

This study primarily aims to develop an antibiotic from the seeds and skin of Pong Pong Cerebra Odollam. Specifically, this seeks to answer the following questions:

1.What are the active constituents present in Pong Pong Cerbera odollam seeds and skin?

2.What is the toxicity level of the active constituents of the Pong Pong Cerbera odollam seeds and skin?

3.What is the effectiveness of the crude extract from Pong Pong Cerbera odollam seeds and skin against selected gram negative and gram positive bacteria?

4. What is the zone of inhibition of the crude extract from Pong Pong Cerbera odollam seeds and skin?

5. Is there significant difference between the antibiotics crude extract from Pong Pong Cerbera odollam seeds and skin and commercial antibiotics?

Hypothesis

Ho = There is no significant difference between the crude extract from Pong Pong seeds and skin and commercial antibiotics

Ha = There is significant difference between the crude extract from Pong Pong seeds and skin and commercial antibiotics

Significance of the Study

Antibiotics have saved more lives since the early 1900’s than almost any other medi-cation. It is essential that we prevent bacteria becoming resistant to antibiotics, because in a world without antibiotics, many, many millions of people would die of infections that we were able to cure when the antibiotic still worked. Good personal hygiene will minimize spread of these bacteria in the community. If the product is proven to be suc-cessful, firms can start plantations, which will provide employment. It increases compe-tition between other antibiotics that are produced globally. New Cerbera odollam plan-tations may provide more employment, which will be beneficial for everyone.

Scope and limitation

The scope of this study is on the preparation of crude extract from Pong Pong seeds and skins as source of antibiotics for gram positive and negative bacteria. This study is limited and will not concentrate on capsulation and administration to humans.

Review of Related Literature

Note that review is omitted to make the banner presentation vey concise. The re-viewed literature was submitted fully to the Science teacher.

Methodology A. Gathering of Materials

Pong-Pong fruits were taken from the school vicinity and the chemicals used for crude extraction were taken from the school laboratory.

B. Methanol Extraction and Waterbath of Pong Pong Fruits Six kilograms of the skin of Pongpong fruits were chopped into smaller pieces and soaked in 1:1 ratio of methanol and water for 15 days. The plant sample was grinded using the blender and the liquid extract was obtained by filtration. The filtrate was collected and subjected to waterbath to recover the crude extract. C. High-Performance Liquid Chromatography

It is a technique in analytical chemistry used to separate, identify, and quantify each component in a mixture. It relies on pumps to pass a pressurized liquid sol-vent containing the sample mixture through a column filled with a solid adsorbent ma-terial. Each component in the sample interacts slightly differently with the adsorbent material, causing different flow rates for the different components and leading to the separation of the components as they flow out the column.

D. Microbiological Assay of Pong Pong Crude Extract Against Gram Positive and Gram

Negative Bacteria An agar plate was prepared from 4.6 g agar and 200 ml of water. The mixture was stirred boiled for 1 minute. The melted agar was poured into the petri dish and covered. The plates were prepared by sterilizing the loop before touching the collection area con-taining the staphylococcus aureus and pseudomonas aeruginosa. The loop was glided over the surface of the agar in zig-zag lines and the cover was returned immediately in-to the plate prior to incubation. After three days the zone of inhibition was taken in each petri dish.

Results and Discussion Based from the results of the microbiological assay, the crude extract from pong pong fruits gave a positive results against staphylococcus aureus and pseudomonas aerugino-sa. The crude extrat gave mean zone of inhibitions of 6.3 cm and 6.09 cm respectively. On the other hand, the commercial antibiotic registered mean zone of inhibitions of 19.6 cm and 5.88 cm for the two samples of bacteria. Although, the crude extract did not outlast the commercial antiobiotic in terms of its efficacy and effectiveness, the crude extract from Pong Pong fruits revealed a feasible source of antibiotic.

Antimicrobial Property of Pong Pong (Cerbera odollam) Fruits Against Gram Positive and Gram Negative Bacteria

James, Darren, Nadya, Diksha

SPA to Pongpong: “Together, we can make a difference for Mother Earth.”