CHAPTER I

INTRODUCTION

Background of the Study

People are prone to bacteria and its other forms. It appears in different forms and places. There are typically 40 million bacterial cells in a gram of soil and a million bacterial cells in a milliliter of fresh water; in all, there are approximately five nonillion (51030) bacteria on Earth, forming a biomass that exceeds that of all plants and animals. There are approximately ten times as many bacterial cells in the human flora as there are human cells in the body, with large numbers of bacteria on the skin and as gut flora. The vast majority of the bacteria in the body are rendered harmless by the protective effects of the immune system, and a few are beneficial. Bacteria are present in most habitats on Earth, growing in soil, acidic hot springs, radioactive waste, water, and deep in the Earth's crust, as well as in organic matter and the live bodies of plants and animals, providing outstanding examples of mutualism in the digestive tracts of humans, termites and cockroaches. These bacteria were prevented by organisms that have antibacterial property. One of these is the Alternanthera sessilis also known as Bunga-Bunga in Philippines. Bunga-bunga plant occurs in wet places throughout the Philippines. The leaves are used as vegetable. Young shoots and leaves are eaten as a vegetable in Southeast Asia. Occasionally it is cultivated for food or for use in herbal medicines. This study is conducted to find a plant that has Antibacterial property that will kills or slows down the growth of bacteria. This is also conducted to prove that Bunga-bunga has antibacterial property that can lessen bacteria to grow more throughout the earth.

Statement of the Problem

This study is conducted to determine the antibacterial activity of the bunga-bunga leaves extract. This study also aims to answer the following: 1. Does the Bunga-bunga plant contain flavonoids? 2. Is the chemical component of the bunga-bunga leaves has the potential to kill or slow down the growth of bacteria? 3. Does bunga-bunga leaves extract contain significant amount of chemical component which slows down reproducing bacteria?

Significance of the Study

An antibacterial is a compound or substance that kills or slows down the growth of bacteria. Today, however, with increased knowledge of the causative agents of various infectious diseases, antibacterial has come to denote a broader range of antimicrobial compounds, including antifungal and other compounds. This study is conducted to find out if Bunga-bunga leaves can be another source of antibacterial. It will surely help the people to live safer to bacteria that cause different kinds of diseases but not only the people but all the living organisms throughout earth. Somehow, on this kind of plant or cheaper source can improve our health conditions due to bacteria and diseases. This study is also conducted to prove that Bunga-bunga plant have antibacterial property that slows down the reproduction of bacteria.

Scope and Limitation

The study is conducted from January, 2012. It was limited to the use of Alternanthera sessilis leaves extract as to determine its antibacterial activity and properties. The Bunga-bunga (Alternanthera sessilis) leaves were bought from Bureau of Plants Industry (BPI) located in San Andres Manila. Phytochemical screening and antibacterial test were used to determine the antibacterial property of the Bunga-bunga plant.

CHAPTER II
Review of Related Literature and Studies
Family : Amaranthaceae

BUNGA-BUNGA
Alternanthera sessilis L.

Other scientific names

Gomphrena sessilis Linn. Illecebrum sessile L. Achyranthes villosa Blanco Illecebrum triandrum Llanos Alternanthera denticulata F.Vill.

Common names
Abisrana (Ilk.) Angelica (Sp.) Aritana (Ilk.); Bilanamanut (Mag.) Bunga-bunga (Tag.) Gogoat (Bon.) Halangbang (If.) Inginga (Ig.) Kapal-kapal (Sul.) Karitana (Bis.) kokoong (Bon.) Lapak-lapak (Sul.) Putputok (Bon.) Sakit (Bon.) Tagtagtu (Bon.) Lianzi cao (China) Sessile joyweed (Engl.)

Bunga-bunga is a spreading or prostrate, erect, more or less branched, glabrous, succulent herb, 0.4 to 1.4 meters high. The ultimate branches are covered with two lines of hairs on the internodes, and the flowering ones are ascending. Leaves are stalkless, simple or pinnately compound, the leaflets elliptic, crenate or obscurely toothed, usually about 2.5 to 7 centimeters long, thick and succulent.

Flowers are minute, paniculate, pendulous, white, and crowded in very short, axillary heads. Sepals are lanceolate, 2 to 2.5 millimeters long. Fruit is dry and flattened, broadly obcordate utricle, containing one seed. It is a very common weed in open, usually wet places throughout the Philippines, in settled areas at low and medium altitudes. Bunga-bunga may be collected throughout the year.

Description Alternanthera sessilis has stems that lie flat and are 1-10 dm long. The leaves are sometimes spear-shaped but mostly elliptic and are 0.3-3cm wide. The petioles are 1-5mm long. The bract are shiny white under the flower's sessile spikes. The sepals are 2.5-3mm long and the uticles are 2-2.3mm long. A. sessilis has a somewhat glistening fruit, that is light-beige yellow. It can reach heights of up to 1 m.

Occurs in: estuarine habitats, riparian zones, ruderal/disturbed, wetlands

Alternanthera sessilis is "a plant of damp places, ditches, wet headlands, roadsides, sometimes and a weed of plantations", and is located from sea level to elevations over 2000 m. It is also found in waste areas, gardens and open fields. Alternanthera sessilis is a pest of sugarcane. It is also a weed of rice in tropical areas, and is also associated with bananas and other cereal crops. According to Tomaino (2006), A. sessilis is an "agricultural weed that invades disturbed wet areas in tropical and subtropical areas of the U.S." Overall, A. sessilis has a "low significant ecological impact. A. sessilis is said to have impacts on liver problems by having hepatoprotective effects. Alternanthera sessilis transfers seeds by wind, water, and also rooting at the stem nodes.

Geographical range Native range: Australia, Asia, Northern Mariana Islands, Federated States of Micronesia, Guam, Palau, Philippines, Soloman Islands, Singapore. Known introduced range: United States, Australia: Norfolk and Christmas Island, Ecuador, Fiji, French Polynesia, Marshall Islands, Nauru, New Caledonia.

Introduction pathways to new locations Horticulture: Alternanthera sessilis can be dispersed through horicultural activities (Maki and Galatowitsch, 2004)

Local dispersal methods On animals (local): The wind blows utricles of Alternanthera sessilis to new locations.

Lifecycle stages During August-October, Alternanthera sessilis produces many utricles that spread with wind and water. Not far from where these were blown, seedlings begin to appear in April. Parts utilized Entire plant; use fresh.

Constituents - High in antioxidant carotene. - Phytochemical studies yielded B-carotene, ricinoleic acid, myristic, palmitic, stearic, oleic and linoleic acids, a-spiraterol, uronic acid, and B-sitosterol. - Study of phenolic and flavonoid content revealed: total crude phenolic content, 1.529; total phenolic acid, 1.404; and flavonoids, 0.370 (values in mg/g dry wt.). - Study of phenolic Properties Slightly tart tasting, cooling, antiphlogistic-analgesic , antidermatosis, antiseptic. Considered febrifugal, galactagogue, abortifacient.

Uses/Edibility Some places, leaves and young shoots eaten as vegetables. In Ceylon, largely eaten as vegetable. In Africa, used for relishes, sauces and soups. Considered a famine plant. Folkloric Poultice of pounded fresh material to sprains, burns and eczema. Carbuncle, erysipelas. Used as a wash for eyes.

 In other cuntries, used for treating acne vulgaris, dyspepsia, liver problems. Decoction used for nausea, vomiting, diarrhea, dysentery, cough, bronchitis and diabetes. In West Tropical Africa, used as poultice for boils. In Sri Lanka, used for cystitis, gonorrhea, snake bite. Also, used by mothers to increase flow of milk. In Ghana, salted decoction used for hematemesis. In India, used for gastrointestinal problems; also as cholagogue, abortifacient and febrifuge. Also, used for snake bites. In Nigeria, for headaches and vertigo. In Taiwan, for hepatitis, bronchitis, asthma.

Others Used for animal feed supplement. Alternanthera sessilis is used as a topical treatment for the common skin problem cane vulgaris. The antioxidant carotene is found in large amounts in the plant. In Southeast Asia young shoots and leaves are ingested as vegetables. In folklore, A. sessilis was used for treating sick individuals. Gayathri et al state that A. sessilis is "used for the treatment of biliousness, dyspepsia associated with sluggish liver, chronic congestion of liver, acute and chronic pyelitis, cystitis, gonorrhea, and strangery and snake bite in Sri Lanka." In India and Sri Lanka, it is used for treatment of gastrointestinal problems. A. sessilis is used as a treatment for headaches and vertigo in Nigeria. It is also used to treat hepatitis, bronchitis, and asthma in Taiwan. A. sessilis is a plant that is purchased for a water garden and over the internet for aquariums. Also, it is bought for its low cost and taste by the people of Sri Lanka. A. sessilis is chopped up when it is fed to ducks and is also fed to pigs. It is used in many different foods in Africa, such as soups, relish, and sauces.

Studies
Antimicrobial / Wound Healing Activities: Study on extract of leaves showed

significant results in different parameters of wound healing. The presence of sterols was also confirmed. Hepatoprotective: A study on the hepatoprotective effects of Taiwanese herb Alternanthera sessilis on liver damage induced by various hepatotoxins showed hepatoprotective effects with a reduction of elevation of SGOT and SGPT . Ionone Derivatives / Antimicrobial: Chloroform extract of dried leaves yielded a mixture of diasteromers of a new ionone derivative with low activities against Pseudomonas aeruginosa and Trichophyton mentagrophytes. Antioxidant: (1) Study showed ethanol extract to have 70% free radical scavenging activity. (2) Study concludes Alternanthera sessilis showed potent radical-scavenging activity and metal ion chelating activity. Antidiabetic: Study of aqueous extract of aerial parts showed significant dosedependent antidiabetic activity with lowering of blood glucose concentration, glycosylated hemoglobin, LDL, total cholesterol. Antibacterial: The antibacterial effect of leaves and internodal-segments derived calli of A. sessilis was evaluated against Proteus vulgaris, Strep pyogenes, B subtilis and Salmonella typhi. the ethanolic extracts of leaves and leaves-derived calli were more effective against the selected bacterial than other solvents.Plants extracts showed antibacterial activity against Gram negative P. aeruginosa and Gram positive Staph epidermis. Hematinic Activity: Study showed that Lupo (A. sessilis) has hematinic activity particularly in iron deficiency anemia. Results suggest AS is a potential drug for augmentation of hemoglobin and serum ferritin in iron deficiency anemia. Antipyretic Activity: Study showed the ethanol extract of aerial parts of Alternanthera sessilis has dose-dependent antipyretic activity. Antibacterial / Antifungal: Plants extracts showed antibacterial activity against Gram negative P. aeruginosa and Gram positive Staph epidermis. Plant also showed antifungal activity against yeasts S. cerevisiae and Candida albicans. In order to suggest methodologies for screening the natural products antibacterial activity, two different qualitative methods were evaluated as follows: agar diffusion test,

employing two different types of reservoirs (filter paper disc impregnated with compound-test and wells in dishes) and bioautographic method (agar diffusion and chromatogram layer). Besides, we discussed the aspects of the microdilution method used for the determination of minimum inhibitory concentration (MIC). Agar diffusion well-variant The bacterial inoculum was uniformly spread using sterile cotton swab on a sterile Petri dish MH agar. Nine serial dilutions yielded concentrations of 100, 80, 60, 40, 20, 10, 5, 2.5, and 1.25 mg/mL for extracts and fractions and four serial dilutions yielded concentrations of 20, 15, 10 e 5 mg/mL for pure substances. 50 L of natural products were added to each of the 5 wells (7 mm diameter holes cut in the agar gel, 20 mm apart from one another). The systems were incubated for 24 h at 36C 1C, under aerobic conditions. After incubation, confluent bacterial growth was observed. Inhibition of the bacterial growth was measured in mm. Reference commercial discs were used (chloramphenicol 30 mg purchased from Cecon and vehicle, 50 mL). Tests were performed in duplicate (18). Agar diffusion disc-variant Natural products were dissolved and diluted with solvents as mentioned previously. Same number of subsequent dilutions was performed as described above. However, natural products serial dilutions were performed out of initial concentrations 2.5 greater than the ones performed for well-variant method (i.e. 250 mg/mL for extracts and fractions and 50 mg/mL for pure substances); 7 mm filter paper discs (Whatman, no. 3) were impregnated with 20 mL of each of the different dilutions. The discs were allowed to remain at room temperature until complete diluent evaporation and kept under refrigeration until ready to be used. Discs loaded with natural products were placed onto the surface of the agar. Commercial chloramphenicol discs(30 mg) and paper discs impregnated with 20 mL of diluents used to dilute natural products were used as control.

Antibacterial Potential of Leaf extract of Bunga-bunga Plant (Alternanthera sessilis)

Rowsette Dorothy D. Llanes IV-Descartes

Submitted in partial fulfillment of the requirements in Research II

Manuel G. Araullo High School Taft Ave. cor. U.N. Ave., Ermita, Manila January, 2012

Ms. Cecile Santiago

(Research Adviser)