production of germicidal soap

PRODUCTION OF GERMICIDAL SOAP WITH TUBA-TUBA (JATROPHA curcas) LEAF STALK EXTRACT

S.Y. 2008-2009

PRODUCTION OF GERMICIDAL SOAP WITH TUBA-TUBA (Jatropha curcas) LEAF STALK EXTRACT

Gretchen Abenoja Dominique Germo Mark Pearl Masing Kathy Junne Matalaba Clark Rustum Sala Dionna Sala Arjay Tolibas

A Research Paper Presented to the Faculty of the Natural Science Area, Arts and Sciences Department SLSU – Tomas Oppus, Southern Leyte In Partial Fulfillment of the Subject Requirements in Science Research

S.Y. 2008-2009

ABSTRACT

The Tuba-tuba (Jatropha curcas) leaf stalk extract was utilized for the production of Germicidal soap. The germicidal soaps were analyzed for sensitivity test. The Kirby-Bauer was used to determine the germicidal soap’s capability in destroying microorganism (bacteria) as indicated in the diameter of the zones of inhibition. Four kinds of bacteria were used for the said test namely. Bacillus subtilis, Staphylococcus aureus, E. coli and Salmonella spp. Three treatments such as 70% Tuba-tuba, and 50% Tuba-tuba, and 30% Tuba-tuba, lather soap and the controls (positive: penicillin, Streptomycin; negative: H 2O) were used to determine if the amount of Tuba-tuba added can affect its potency against the bacteria. The result showed that 70% Tuba-tuba leaf stalk extract was the best germicidal soap in killing Staphylococcus aureus and Bascillus subtilis only not in E. coli and Salmonella spp. And streptomycin is better in killing E. coli and Salmonella spp.

INTRODUCTION

Nature and Importance of the Study Tuba – tuba (Jatropha curcas) also known as Tubang Bakod in Tagalog, Physic Nut in English or interchangeably Tuba – tuba or Jatropha is one of the most promising source of bio – fuel today. About 30% of Tuba – tuba nut is composed of oil. The Jatropha plant’s average height is about three meters, so harvesting is easy and plant can be grown practically anywhere (ordinary soil, sandy, gravely or rocky soil) and adapts easily to different climates. Jatropha is resistant to droughts. It can stand up to droughts. It can stand up to two years without rainfall. The tree also a short gestation period; it will bear several fruits about six months old and be fully bearing between one to two years (http://www.Philippinesherbalmedicine.org/tuba-tuba.html). Skin disease is a very broad term that describes numerous conditions. Some skin diseases are serious and deadly, while others are just annoying. Some

skin

diseases

are

disfiguring,

while

others

are

barely

visible

(http://dermatology.about.com/oldskindiseases/a/skindisease.htm) Germicidal soap is used to eliminate germs in the body. It also removes skin germs that may cause skin infection and perspiration odor. It prevents infection by inhibiting the growth of bacteria. This research concentrates on testing the capability of the produced soap on bacteria such as Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Salmonella spp.

Objectives of the Study The research study aimed to: 1. make a germicidal soap with the presence of Tuba – tuba leaf stalk extract; 2. determine the germicidal soap’s capability in destroying microorganism (bacteria); and 3. compare treatments to determine the best germicidal soap in terms of the following: a. color; b. odor; c. texture; d. ability to destroy microorganism (bacteria)

Scope and Limitation of the Study This study focused on getting the extract from the leaf stalk of the Tuba – tuba (Jatropha curcas) plant; making a germicidal soap with the presence of the Tuba – tuba leaf stalk extract; comparing the finished products with each treatment against cultured bacteria to identify which is the best germicidal soap among the treatments.

Time and Place of the Study This study was conducted at Southern Leyte State University – Tomas Oppus (SLSU – TO) where the germicidal soaps were made and at Visayas State University (VSU) where the testing of the soap’s effectiveness against bacteria such as: Bacillus subtilis, Staphylococcus aurues, E. coli and Salmonella spp.

REVIEW OF RELATED LITERATURE Related Literature Tuba–tuba (Jatropha curcas) is a non-edible plant that grows mostly in the tropical countries like the Philippines. It is resistant to drought and can easily be planted or propagated through seeds or cuttings. It starts producing seeds within 14 months, but reaches its maximum productivity level after 4-5 years. The plant remains useful for around 30-40 years (jgj.doe.gov). The hardy Jatropha is resistant to pests and produce seeds containing up to 40% oil. When the seeds are crushed and processed, the resulting oil can be used in a standard diesel engine, while the residue can also be processed into biomass to power electricity plants. Estimates of Jatropha seed yield vary wildly, due to lack or research data, the genetic diversity of the crop, the range of environments in which it is grown, and Jatropha’s perennial life cycle. Seeds yields under cultivation can range from 1,500 to 2,000 kilograms per hectare, corresponding to extractable oil yields of 540 to 680 liters per hectare. Jatropha can also be intercropped with other cash crops such as coffee, sugar, fruits and vegetables (http://www.limbopascal07.blogspot.com). Physic nut (Jatropha curcas or tuba-tuba) is a member of the family euphorbiaceous. Physic Nut, large shrub or small tree of the spurge family, native to tropical America. The plant grows to 4.5 m (15 ft), bearing roundish,

lobed leaves up to 15 m (6 in) wide and clusters of yellow flowers. The fruit, small grayish nut, contains a white, edible kernel, which is the source of Curcas oil. This oil is used in cooking, soap making, and as an illuminant. Staphylococci (staph) are Gram-positives spherical bacteria that occur in microscopic clusters resembling grapes. Bacteriological culture of the nose and skin of normal humans invariably yields staphylococci. In 1884, Rosenbach described the two pigmented colony types of staphylococci and proposed the appropriate nomenclature: Staphylococcus aureus (yellow) and Stapylococcus albus (white). The latter species is now named Staphylococcus epidermis. Although more than 20 species of Staphylococcus are described in Bergey’s Manual (2001), only Staphylococcus aureus and Staphylococcus epidermis are significant in their interactions with humans. S. aureus colonizes mainly the nasal passages, but it may be found regularly in most other anatomical locales, including the skin, oral cavity and gastrointestinal tract. Taxonomically, the genus Staphylococcus is in the Bacterial family Staphylococcaceae, which includes three lesser known genera, Gamella, Macrococcus and Salinicoccus. The best-known of its nearby phylogenetic relatives are the members of the genus bacillus in the family Bacillaceae, which is on the same level as the family Staphylococcaceae. The Listeriaceae are also a nearby family Staphylococcus aureus forms a fairly large yellow colony on rich medium; S. aureus is often hemolytic on blood agar. S. aureus an grow at a temperature range of 15 to 45 degrees and at NaCl concentration as high as 15 percent. Theodor Escherich first described E. coli in1885, as Bacterium coli

commune, which he isolated from feces of newborns. It was later renamed Escherichia coli, and for many years the bacterium was simply considered to be, a commensal organism of the large intestine. It was not until 1935 that a strain of E. coli was shown to be the cause of an outbreak of diarrhea among infants. The GI tract of most warm-blooded animals is colonized by E. coli within hours or a few days after birth. The bacterium is ingested in foods or water or obtained directly from other individuals handling the infant. The human bowel is usually colonized within 40 hours of birth. E. coli can adhere to the mucus overlying the large intestine. Once established, an E. coli strain may persist for months or years. Resident strains shift over a long period (weeks to months), and more rapidly after enteric infection or antimicrobial chemotherapy that perturbs the normal flora. The basis for these shifts and the ecology of Escherichia coli in the intestine of humans are poorly understood despite the vast amount of information on almost every other aspect of the organism’s existence. The entire DNA base sequence of the E. coli genome has been known since 1997. E. coli is the head of the large bacterial family, Enterobacteriaceae, the enteric bacteria, which are facultatively anaerobic Gram-negative rods that live in the intestinal tracts of animals in health and disease. A Physiologically, E. coli is versatile and well-adapted to its characteristic habitat. It can grow in media with glucose as the sole organic constituent. Wildtype E. coli has no growth factor requirements, and metabolically it can transform glucose into all of the macromolecular components that make up the cell. The bacterium can grow in the presence of absence of O 2.

E. coli can respond to environment signals such as chemicals, pH, temperature, osmolarity, etc., in a number of very remarkable ways considering it is a unicellular organism. For example, it can sense the presence or absence of chemicals and gases in its environment and swim towards or away from them. Or it can stop swimming and grow fimbriae that will specifically attach it to a cell or surface receptor. In response to change in temperature and osmolarity, it can vary the pore diameter of its outer membrane porins to accommodate larger molecules (nutrients) or to exclude inhibitory substances. With its complex mechanisms for regulation of metabolism the bacterium can survey the chemical contents in its environment in advance of synthesizing any enzymes that metabolize these compounds. It does not wastefully produce enzymes for degradation of carbon sources unless they are available, and it does not produce enzymes for the synthesis of metabolites if they are available as nutrients in the environment. E. coli is a consistent inhabitant of the human intestinal tract, and it is the predominant facultative organism in the human GI tract; however, it makes up a very small proportion of the total bacterial content. The anaerobic Bacteroides species in the bowel outnumber E. coli by at least 20:1. However, the regular presence of E. coli in the human intestine and feces has led to tracking the bacterium in nature as an indicator of fecal pollution and water contamination. As such, it is taken to mean that, wherever E. coli is found, there may be fecal contamination by the intestinal parasites of humans.

Salmonella is a Gram-negative facultative rod-shaped bacterium in the same proteobacterial family as Escherichia coli, the family Enterobacteriaceae, trivially known as “enteric” bacteria. Salmonella is nearly as well-studied as E. coli from a structural, biochemical and molecular point of view, and as poorly understood as E. coli from an ecological point of view. Salmonellae live in the intestinal tracts of warm and clod blooded animals. Some species are ubiquitous. Other species are specifically adapted to a particular host. In humans, Salmonella are the cause of two diseases called salmonellosis: enteric fever (typhoid), resulting from bacterial invasion of the bloodstream, and acute gastroenteritis, resulting from a forborne infection/intoxication. The genus Salmonella is a member of the family Enterobacteriaceae It is composed of bacteria related to each other both phenotypically and genotypically. Salmonella DNA base composition is 50-52 mol% G+C, similar to that of Escherichia, Shigella, and Citrobacter. The bacteria of the genus Salmonella are also related to each other by DNA sequence. The genera with DNA most closely related to Salmonela are Escherichia, Shigella, and Citrobacter. Similar relationships were found by numerical taxonomy and 16S ssRNA analysis. Bacillus subtilis, known as the hay bacillus or grass bacillus, is a Grampositive, catalase-positive bacterium commonly found in soil. A member of the genus Bacillus, B. subtilis is rod-shaped and has the ability to form a tough, protective endospore, allowing the organism to tolerate extreme environmental conditions. Unlike several other well-known species, B. subtilis has historically

been classified as an obligate-aerobe, though recent research has demonstrated that this is not strictly correct. B. subtilis is not considered a human pathogen; it may contaminate food but rarely causes food poisoning. B. subtilis produces the proteolytic enzyme subtilisin. B. subtilis spores can survive the extreme heating that is often used to cook food, and it is responsible for causing ropiness – a sticky, stringy consistency caused by bacterial production of long-chain polysaccharides – in spoiled bread dough. B. subtilis can divide asymmetrically, producing an endospore that is resistant to environmental factors such as heat, acid, and salt, and which can persist in the environment for long periods of time. The endospore is formed at times of nutritional stress, allowing the organism to persist in the environment until conditions become favorable. Prior to the process to produce the spore the bacterium might become motile, through the production of flagella, and also take up DNA from the environment. Soap, cleansing agent or detergent is made from animal and vegetable fats, oils, and greases; chemically sodium or and oil with alkali. Hard soap are made from oils and fats that contain a high percentage of saturated acids, which are

saponified

with

sodium

hydroxide

(http://www.textbookofbacteriology.net/staph.html). Antibacterial soap is any cleaning product to which active antibacterial ingredients have been added. These chemicals kill bacteria and microbes. They do not kill viruses.

Many, or even most, liquid hand and body soaps contain antibacterial chemicals. Triclosan is a common ingredient, as is alcohol. Since there is a great variety of bacteria, effectiveness against any given type of bacterium does not ensure that it is effective against unrelated types. These are generally only contained at preservative level unless the product is marked antibacterial, antiseptic, germicidal, or deodorant. Triclosan, Triclocarban/Trichlorocarbamide and PCMX/Choloroxylenol are commonly used for antibacterial and deodorant effect in consumer products. Some soap contains tetra sodium EDTA which is a chelating agent that sequesters metals that the bacteria require in order to grow. Other microbes also require metals and so it is actually anti-microbial agent that is widely used even as a preservative. It appears to be fairly harmless in the environment.

Related Studies Tuba-tuba (Jatropha curcas) is a viable source of biodiesel in our country. There are benefits in planting tube-tuba (Jatropha curcas) aside from using the seed oil as biodiesel. The extracted oil can be also used in making soap and the leaves can be used as yellow dye while the bark extract as blue dye. The seeds when pounded can be used tanning. Its roots, flowers and later are said to have medicinal properties. Planting Tuba-tuba (Jatropha curcas) reduces oil degradation, erosion and deforestation of the countryside. There is also a possibility that the community could also extract Jatropha fuel without desertification, and directly use the extraction technology however

has yet to be assessed. Further study is also being done on this process and its impacts to health, as the toxicity of its emission has yet to be determine (jgl.doe.gov) Last April 11, 2008 the tuba-tuba or Jatropha was studied and was proven to be effective as a liniment for stomachache. This was made to help poor people that could not afford buying medicines that would fit their budget. It has no harmful

effects

and

can

be

good

substitute

for

the

leading

band

(http://www.limbopascalo7.blogspot.com) Jatropha curcas, while it is no-edible, is known as an oil-yielding perennial shrub with multiple use. Whole seeds are reported as being purgative and have insecticidal and fungicidal properties. The latex is used in healing wounds and in the treatment of various skin problems, decoction of the roots is known to are indigestion and diarrhea, the back as poultice for sprains and dislocations, fresh stems as toothbrushes and to strengthen gums and are bleeding, spongy gums or gum-boils. Jatropha has many other known medicinal uses extracts have been shown to have anti-tumor activity, latex contains alkaloid jatrophine which shows anti cancer properties, seed can be used as remedy for constipation, and leaves are boiled to cure malaria. In the Philippines, jatropha leaves are used to cover swelling portion of the body and the bark is known to cure dislocated bones. Seed composition at 6.2% moisture content, protein is 18%, fat 38%, carbohydrates 17%, fiber 15.5%, ash 5.3%. The oil content is 25-30% seeds, 5060% in kernel: oil contains 21% saturated fat and 79% unsaturated fat. Oil has

high saponification value (195.0). Thus, it is excellent for soap making (Heller, Joachim. 1996) Fat or oil and alkali taken from lyre are the basic ingredients of soap. Fat serves as the foundation of the soap and alkali produces the chemical reactions that makes the mixture hard and gives it cleaning ability.

MATERIALS AND METHODS

Research Design This study utilized RCBD or Randomized Complete Block Design. There were ten replicates in each treatment.

Research Respondents Respondents were selected by random sampling. The selected 30 first year to third year laboratory high school students of SLSU-TO were the respondents of this study. There were ten selected students in each year level to meet all the thirty respondents.

Sampling Technique This study used the simple random sampling in which all the members had a chance to be selected. This was done by simply writing the names of all members of the population in rolled pieces of papers and placing them in a container. The 30 respondents were picked out from the container.

Research Instrument This study utilized a self-made questionnaire that supported the objectives of the study. The Part I of the self-made questionnaire described the respondents profile and the Part II is the evaluation of the germicidal soap in terms of odor, texture and color.

Gathering of the Tuba-tuba Leaf Stalk Tuba-tuba leaves were gathered, washed thoroughly, and air dried.

Extraction of the Tuba-tuba Leaf stalks The leaf stalks were separated from the leaves and were cut into pieces by using a clean knife. After that, the juice from the leaf stalk was extracted through pounding with the use of mortar and pestle.

Preparation of Treatments Tuba-tuba leaf Treatments

% Concentration

Distilled Water stalk extract

T1

70% Tuba-tuba

70 ml

30 ml

T2

50% Tuba-tuba

50 ml

50 ml

T3

30% Tuba-tuba

30 ml

70 ml

Control

Content

T0 +

Antibiotic (Penicillin and Streptomycin)

T0 -

Water

Saponification of Fatty Acids Fifteen ml of treatment 1 with 40 ml distilled water was poured into a 500 ml beaker. While doing this, 9.6 grams of KOH was mixed with 40 ml distilled water placed in a 250 ml beaker. KOH had been dissolved by stirring the solution constantly. Then, a saturated salt-water solution was prepared by putting table salt constantly in 40 ml distilled water. And 40 ml distilled water with 40 ml Sun Valley oil was prepared. All the prepared solutions were mixed.

The solution was heated with constant stirring until sticky. In the process, another 9.6 grams of KOH with 40 ml distilled water and 40 ml saturated salt solution were added (see plate 1, Appendix G). A droplet of the mixture was placed into a beaker with tap water. If the tap water could no longer have droplets of oil on top, this means that the saponification of soap was done. If there were still droplets of oil, heating and stirring were continued.

Purification of Soap The beaker was removed from the steam boiler and the contents were poured into a 250 ml of saturated salt-water solution using tap water. The solution was filtered using chess cloth. The chess cloth was pressed and the solidified soap was placed in a beaker with 20 ml distilled water. The soap was poured into a cupboard mold and let it cool down (see plate 2, Appendix G). The soap was tested by using it to the hands. It was proven that the soap was made because it lathered. It was left in a secured place where no moisture could enter. The next day, it has already hardened. All the procedures were repeated for the other treatments.

Sensitivity test of the Produced Soap A modified Kirby-Bauer method for sensitivity test was employed in this study to assess the bactericidal activity which was indicated by zones of inhibition (Coles, 1980). The zone of inhibition is the area on an agar plate where growth of a control organism is prevented by an antibiotic usually placed or impregnated on the agar surface (see plate 3, Appendix G). Sensitivity testing was done on pure culture of isolated organisms. Sterile cotton swab was soaked in the prepared suspension of the test organisms and was swabbed on the entire surface of the agar. The inoculated plates were then allowed to dry on the bench. After the plates had completely dried, five plastic wells were placed in each Petri plate based on the devised pattern. Four plates were allotted in each test organism. Three plastic wells were treated with three droplets of each treatment and a single plastic wells treated with the three controls (H2O), and along the positive control (+streptomycin and +penicillin) were impregnated in each plate. These were done on all test organisms using the treatments. The preparation was labeled and incubated at 37°C for 24 hours. The zone of inhibition was measured from the edge of the plastic wells to the perimeter, using a metric ruler held near the surface of the medium (see plate 4, Appendix G).

Statistical Design Each of the three experimental groups (treatments 1, 2, 3) was compared to the control group (treatment 0) using t-test to determine which among the different treatments produced the best germicidal soap. The comparison (between a treatment and the control) which produced the least p-value was considered the best germicidal soap.

Extraction of Tubatuba leaf stalks

Preparation of Percent Concentration

Saponification of Fatty Acid

Purification of Soap

Sensitivity Test

Data Gathering

Data Analysis

Figure 1. Flow Chart of the Study

RESULTS AND DISCUSSION

Table 1. Mean of the zone of inhibition of the three treatments against Staphylococcus aureus, Bacillus subtilis, E. coli and Salmonella spp. Treatments Staphylococcu Bacillus E. coli Salmonella s Subtilis (mm) Spp. Aureus (mm) (mm) (mm) T0-distilled NZ NZ NZ NZ Water T0+strep NZ NZ 29.3 24.8 T0+penicillin 18.7 18 NZ NZ T1- 30% 14.5 14.6 18.2 18.6 T2- 50% 15 12 19.45 19.6 T3- 70% 21.5 17.2 21.7 20 Based on the table shown above, the zones of inhibition of the plastic wells with 70% of Tuba-tuba leaf stalk extract were greater than the zones of inhibition of distilled water, streptomycin, penicillin, 30% Tuba-tuba leaf stalk extract and 50% Tuba-tuba leaf stalk extract. This indicates that the treatment three (3) which had 70% Tuba-tuba leaf stalk extract was the best germicidal soap in terms of its ability to destroy microorganism (bacteria).

Table 2. Mean of the responses in terms of color of the three (3) germicidal soaps Treatments Weighted Mean Level of Acceptability Soap 1 2.73 Very Satisfactory (T1 – 30% Tuba-tuba leaf stalk extract) Soap 2 2.80 Very Satisfactory (T2 – 50% Tuba-tuba leaf stalk extract) Soap 3 2.80 Very Satisfactory (T3 – 70% Tuba-tuba leaf stalk extract)

Legend: Mean Rating 2.34-3.00 1.67-2.33 1.00-1.66

Color White Dirty White Dull or Dim

Descriptive Rating Very Satisfactory Satisfactory Needs Improvement

As shown, the three germicidal soaps were identified as very satisfactory which means that the three germicidal soaps did not differ in terms of its color.

Table 3. Mean of the responses in terms of texture of the three (3) germicidal soaps Treatments Weighted Mean Level of Acceptability Soap 1 2.50 Very Satisfactory (T1 – 30% Tuba-tuba leaf stalk extract) Soap 2 2.40 Very Satisfactory (T2 – 50% Tuba-tuba leaf stalk extract) Soap 3 2.37 Very Satisfactory (T3 – 70% Tuba-tuba leaf stalk extract) Legend: Mean Rating 2.34-3.00 1.67-2.33 1.00-1.66

Texture Smooth Less Smooth Rough

Descriptive Rating Very Satisfactory Satisfactory Needs Improvement

It is observed that the three germicidal soaps were found out as “very satisfactory” by the respondents.

Table 4. Mean of the responses in terms of odor of the three (3) germicidal soaps Treatments Weighted Mean Level of Acceptability Soap 1 2.07 Satisfactory (T1 – 30% Tuba-tuba leaf stalk extract) Soap 2 2.10 Satisfactory (T2 – 50% Tuba-tuba leaf stalk extract) Soap 3 1.72 Satisfactory (T3 – 70% Tuba-tuba leaf stalk extract) Legend: Mean Rating 2.34-3.00 1.67-2.33

Odor Fragrant Less Fragrant

Descriptive Rating Very Satisfactory Satisfactory

1.00-1.66

Smelly

Needs Improvement

As shown, Soap 1, Soap 2, and Soap 3 were considered “satisfactory” by the high school students.

Table 3. Mean of the responses in terms of texture of the three (3) germicidal soaps Treatments Weighted Mean Level of Acceptability Soap 1 2.43 Very Satisfactory (T1 – 30% Tuba-tuba leaf stalk extract) Soap 2 2.43 Very Satisfactory (T2 – 50% Tuba-tuba leaf stalk extract) Soap 3 2.29 Satisfactory (T3 – 70% Tuba-tuba leaf stalk extract) Legend: Mean Rating 2.34-3.00 1.67-2.33 1.00-1.66

Descriptive Rating Very Satisfactory Satisfactory Needs Improvement

Based on table 5, Soap 1 and Soap 2 were very satisfactory while Soap 3 was satisfactory in terms of its overall acceptability.

Table 6. Comparison of the acceptability of the germicidal soaps in terms of color Variables Weighted Mean P – Value Decision on Ho S1

S2

S1 vs. S2

2.73

2.80

S1 vs. S3

2.73

S2 vs. S3

2.80

S3 0.42350

Accepted Ho

2.80

0.48884

Accepted Ho

2.80

1.00000

Accepted Ho

a= 0.05 As shown, the probability values of S1 vs. S2, S1 vs. S3 and S2 vs. S3 were equal and greater than the set level of rule of significance of 0.05, accepting the hypothesis which means the germicidal soaps treatments did not differ terms of its color. Table 7. Comparison of the acceptability of the germicidal soaps in terms of textures Variables Weighted Mean P – Value Decision on Ho S1

S2

S1 vs. S2

2.50

2.40

S1 vs. S3

2.50

S2 vs. S3

2.40

S3 0.54070

Accepted Ho

2.37

0.45906

Accepted Ho

2.37

0.82297

Accepted Ho

a= 0.05 As shown in Table 7, the probability values of S1 vs. S2, S1 vs. S3 and S2 vs. S3 were equal or greater than 0.05, accepting the hypothesis which says that the germicidal soaps did not differ in terms of its texture.

Table 8. Comparison of the acceptability of the germicidal soaps in terms of odor Variables Weighted Mean P – Value Decision on Ho S1

S2

S1 vs. S2

2.07

2.10

S1 vs. S3

2.07

S2 vs. S3

2.10

S3 0.82297

Accepted Ho

1.70

0.03173

Reject Ho

1.70

0.00297

Reject Ho

a= 0.05 Furthermore, the probability values for S1 vs. S2 is greater than 0.05, thus accepting the hypothesis which means that odors did not differ among the two germicidal soaps. On the other hand, a significant differed between the two germicidal soaps.

Table 9. Comparison of the acceptability of the germicidal soaps in terms of odor Variables Weighted Mean P – Value Decision on Ho S1

S2

S1 vs. S2

2.43

2.43

S1 vs. S3

2.43

S2 vs. S3

2.43

S3 1.00000

Accepted Ho

2.29

0.02937

Reject Ho

2.29

0.02937

Reject Ho

a= 0.05 Finally, the probability value of S1 vs. S2 was greater than 0.05, accepting the hypothesis which means that the two germicidal soaps did not differ in terms of its overall acceptability. On the other hand, a significant difference was observed among S1 vs. S3 and S2 vs. S3. This shows that the germicidal soaps differed in terms of overall acceptability.

SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS

Summary The research work was conducted at Southern Leyte State UniversityTomas Oppus (SLSU-TO) and Visayas State University (VSU). The objectives of the study were to make a germicidal soap with the presence of the Tuba-tuba (Jatropha curcas) leaf-stalk extract; to determine the germicidal soap’s capability in destroying microorganism (bacteria); compare treatments to the best germicidal soap in terms of the color, odor, texture and ability to destroy microorganism (bacteria). A modified Kirby-Bauer for sensitivity test was employed in this study to assess the germicidal activity of the produced soap as indicated by the diameter of the zones of inhibition. There were three treatments; 30% Tuba-tuba, 50% Tuba-tuba, 70% Tuba-tuba. The controls were the positive control (water), and negative control (streptomycin and penicillin). The mean diameters of the zone of inhibition of the treatments were computed using t-test. The greater was the diameter of inhibition, the more effective was the soap. It means it could kill more bacteria. This research activity was successful in producing a soap which could kill certain bacteria.

Conclusion The extract of the Jatropha curcas Leaf Stalk can be utilized for soap production. Based on the results gathered, it was found out that the germicidal soap from Jatropha curcas leaf stalk extract was effective in killing various bacteria most especially the Bacillus subtilis and the Salmonella. Among the three treatments, treatment 70% Jatropha curcas leaf stalk extract was the best treatment in killing or destroying the microorganism.

Recommendation Based on the findings and the results gathered, it is recommended that a more comprehensive methodology should be observed in getting the extract and in the production of the soap. It is suggested to test the effectiveness of the germicidal soap from the tuba-tuba leaf stalk directly to animals having skin disease since it is a standard operating procedure in research to test the product first on animals before on humans.

BIBLIOGRAPHY A. BOOKS •

Webster Dictionary/Thesaurus

•

Diksyunaryong Filipinos-Inges

B. ELECTRONIC SOURCES •

http://dermatology.about.com/oldskindiseases/a/skindisease.html

•

http://www.limbopascal07.blogspot.com

•

http://www.philippinesherbalmedicine.org/tuba-tuba.html

•

http://textbookofbacteriology.net/staph.html

•

jgj.doe.gov

•

www.altavista.com

•

www.ask.com

•

www.google.com

•

www.soapcrone.com

C. UNPUBLISHED THESIS/BOOKS •

Efficacy of Bitaog (Calophyllum inophyllum) Soap Against Akin DiseaseCausing Bacteria

Sensitivity test of Jatropha spp.plant extract against gram negative and gram positive microorganism Treatments T0-dist. Water 1 2 3 4 5 6 7 8 9 10 T0 +strep. 1 2 3 4 5 6 7 8 9 10 T0 +penicillin 1 2 3 4 5 6 7 8 9 10 T1-30% 1 2 3 4 5 6 7 8 9 10

Staph. Aureus

Bacillus subtilis

E. coli

Salmonella spp

NZ* NZ NZ NZ NZ NZ NZ NZ NZ NZ

NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

31 30 31 30 30 25 30 25 30 31

25 28 30 30 15 25 25 25 25 20

16 20 20 24 13 16 13 16 20 24

20 20 20 14 20 17 25 15 15 20

10 15 12 15 20 25 15 25 25 25

15 15 20 15 15 15 15 25 25 20

13 10 12 17 10 20 20 14 9 20

12 10 11 20 20 15 13 15 15 15

Treatments T2-50% 1 2 3 4 5 6 7 8 9 10 T3-70% 1 2 3 4 5 6 7 8 9 10 Lather soap 1 2 3 4 5 6 7 8 9 10

Staph. Aureus

Bacillus subtilis

E. coli

Salmonella spp

16 12 11 15 11 15 15 20 20 15

10 16 15 12 10 8 14 10 15 10

14 16 15 25 25 17 20 20 25 17.5

20 25 19 15 20 15 25 17 20 20

20 20 17 20 25 18 25 25 25 20

20 17 15 20 15 15 20 20 15 15

14 19 24 25 23 22 25 25 23 17

20 20 25 25 20 20 25 15 15 15

15 20 25 10 25 15 15 18 17 17

25 25 20 19 15 20 20 20 20 20

31 30 30 31 30 30 31 30 25 25

21 15 20 20 15 15 15 21 20 20

*NZ= No Zone of inhibition or 0

MA. DELIA AMIHAN-PAGENTE Sci. Res. Asst. EUGENE B. LANADA, DVM., MPhil Dean, College of Veterinary Medicine

Zones of inhibition of the negative control (water0 against Staphylococcus aureus, Bacillus subtilis, E.coli and Salmonella spp. TO- dist. Staph. Bacillus E. coli Salmonella water 1 2 3 4 5 6 7 8 9 10

aureus NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

Subtilis NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

Spp. NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

Zones of inhibition of the positive control (streptomycin) against Staphylococcus aureus, Bacillus subtilis, E.coli and Salmonella spp. Staph. Bacillus E. coli Salmonella TO+strep. aureus Subtilis (mm) Spp. 1 2 3

NZ NZ NZ

NZ NZ NZ

31 30 31

(mm) 25 28 30

4 5 6 7 8 9 10 Mean

NZ NZ NZ NZ NZ NZ NZ

NZ NZ NZ NZ NZ NZ NZ

30 30 25 30 25 30 31 29.3

30 15 25 25 25 25 20 24.8

Zones of inhibition of the other control (penicillin) against Staphylococcus aureus, Bacillus subtilis, E.coli and Salmonella spp. TO+ Staph. Bacillus E. coli Salmonella penicillin

aureus

Subtilis

1 2 3 4 5 6 7 8 9 10 Mean

(mm) 10 15 12 15 20 25 15 25 25 25 18.7

(mm) 15 15 20 15 15 15 15 25 25 20 18

Spp. NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

NZ NZ NZ NZ NZ NZ NZ NZ NZ NZ

Zones of inhibition of the treatment using 30% of Tuba-tuba leaf-stalk extract against Staphylococcus aureus, Bacillus subtilis, E.coli and Salmonella spp. Staph. Bacillus E. coli Salmonella T1-30% aureus Subtilis (mm) Spp. 1 2 3 4 5 6 7 8 9 10 Mean

(mm) 13 10 12 17 10 20 20 14 9 20 14.5

(mm) 12 10 11 20 20 15 13 15 15 15 14.6

16 20 20 24 13 16 13 16 20 24 18.2

(mm) 20 20 20 14 20 17 25 15 15 20 18.6

Zones of inhibition of the treatment using 50% of Tuba-tuba leaf-stalk extract against Staphylococcus aureus, Bacillus subtilis, E.coli and Salmonella spp. Staph. Bacillus E. coli Salmonella T1-50% aureus Subtilis (mm) Spp. 1 2 3 4 5 6 7 8 9 10 Mean

(mm) 16 12 11 15 11 15 15 20 20 15 15

(mm) 10 16 15 12 10 8 14 10 15 10 12

14 16 15 25 25 17 20 20 25 17.5 19.45

(mm) 20 25 19 15 20 15 25 17 20 20 19.6

Zones of inhibition of the treatment using 70% of Tuba-tuba leaf-stalk extract against Staphylococcus aureus, Bacillus subtilis, E.coli and Salmonella spp. Staph. Bacillus E. coli Salmonella T1-70% aureus Subtilis (mm) Spp. 1 2 3 4 5 6 7 8 9 10 Mean

(mm) 20 20 17 20 25 18 25 25 25 20 21.5

(mm) 20 17 15 20 15 15 20 20 15 15 17.2

14 19 24 25 23 22 25 25 23 17 21.7

(mm) 20 20 25 25 20 20 25 15 15 15 20