However, it also is the cause of wound infections and has the potential to induce certain diseases, leading to infections in any of the major organs of the body. It also is responsible for many serious community- and uneconomically- acquired infections, Ewing the most frequently isolated bacterial pathogen from patients with hospital- acquired infections. Although Antibacterial agents are now available in the market, it cannot be denied that these medical preparations are highly expensive and it has side effects that can sometimes prove to be more difficult to manage than the ailment they are meant to cure.
To other researchers who might be investigating the inhibitory activity of the same plants using another biological effect as determinant or using another plant, the effectiveness of protocols adapted to this study may serve s a blueprint for their own experimentation. This study indemnifies that the five be used in the formulation of new herbal preparations as an alternative medication against infections caused by S. erasure. The value of this research to this country, the Philippines, is high.
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Second, this study solely focused on testing which herbal plant extract shows greater, lesser or no inhibitory effect against S. erasure coagulate production. Only the measurement of the inhibition of plant extracts on the coagulate production in S. erasure were studied here. The data of this in vitro experiment was based on the lumping factor of S. erasure. The activities for this study, specifically in the Pharmacy Dispensing Laboratory and Medical Technology Laboratory, where considered necessary materials and apparatus are available to conduct the study within the premises of SST.
Scholastics College of Tactical from the period of May 2011 to February 2012. REVIEW OF RELATED LITERATURE Botanical Description and Economic Importance of Herbal Plants Plumbers Aruba commonly known as “Caliches”, is a succulent shrub or small tree, sometimes with a fleshy taproot. It’s also known as “Frangipani” or “Temple Flower”. It is a member of the family Apocalypse. It’s stem swollen at the base up to 1-mm in diameter. The bark appears pale grayish-green, grey or brown, smooth, with sticky, clear or white latex, leaves arranged spirally, clustered at the end of branches and flowers usually appear before the leaves. EDEN, 2010). The Spanish regime, through the Acapulco trade, brought additional exotic tree species, mostly agricultural crops and that includes caliches. P. Aruba is a native of Mexico and is widely cultivated in the Philippines (Bake et al. , 2005). P. Aruba, aside from it is nearly planted for ornamental purposes; it has several medicinal applications used as a poultice to boils, swelling, and eruptions of the soles of the feet and are reported to be effective for cure of asthma (Reign et al. , 2001). Certain studies on P. Aruba reveal its possible application in various of fields.
A study was shown that extraction of the crude plant contains iridous that have been reported to have antibacterial, illogical, exotic, and/or plant growth inhibitory activity. (Sarandon et al. , 1990). Pomes aquatic is a semi-aquatic tropical plant that’s a popular vegetable in the Philippines. L. Aquatic is commonly known as “Kayaking”, or “Water Spinach”. It is a member of the family Convalescence and described as herbaceous trailing vine with milky sap. Its’ stems are hollow, rooting at nodes, floating in aquatic situations, with up to 3 m long trailing vine. L. Aquatic leaves appeared to be alternate, simple, with glamorous petioles 3-14 CM (1-6 in) long with blades generally arrowhead shaped but variable, to 17 CM (7 in) long, with tips pointed and it’s blades held above water when stems are floating (Ramey et al. , 2007). Only a very few scientific studies have been conducted on its medicinal aspects. These include the inhibition of effects on liver diseases and constipation (Sparkman et al. , 2008). It was found that l. aquatic exerted a higher magnitude of antimicrobial activity against the tested four types of bacterial species namely Escherichia coli, Pseudonymous organisms, S. Rues and Microcosmic lutes than that of the rest two herb extracts Hagiography and Anhydride due to compositional variation in the active bimolecular of three herbs (Major et al. , 2009). Euphorbia hart belongs to the family Euphorically. It is a small annual herb common to tropical countries (Sparrow, 1982). E. hart, commonly known as “Gates-Gates”, can grow to a height of 40 CM. The stem is slender and often reddish in color, covered with yellowish bristly hairs especially in the younger parts. The leaves are oppositely arranged and are usually greenish or reddish underneath measuring about 5 CM long.
In the axial appear very small dense round clusters of flowers. The stem and leaves produce white or milky Juice when cut (Lind et al. , 1971). Earlier, objectivity studies describe E. hart L. As a potent medicinal plant and established its sedative and nationally activity, analgesic, antipathetic, anti- inflammatory, antidepressant for blood pressure, intransitives and antioxidants. The ethanol extract of the leaves of E. hart L. Was analyzed for their antimicrobial activity by agar well diffusion method against Gram positive bacteria species including S. erasure (Oar et al. , 2010). Leaves of E. Art were used in traditional medicine for the treatment of boils, wounds and control of diarrhea and dysentery was extracted by maceration in ethanol (Geeks et al. , 2007). Mimosa podia commonly known as “Mikhail”, “humble plant”, or “touch-me-not”, is a member of the family Moonscape. It is a shrub-like plant, growing to a height of about CACM. The stem is upright, much branching and sparsely coated with fine, white hairs; and thus the plant has a prickly feel. The leaves are pinnate and these leaflets are called pinnate, and these are sub-divided into many little leaflets called penniless, thus giving the plant a fern-like appearance.
The pinnate are 2. 5 to CM long and are made up of elliptical, 0. CM long penniless arranged in rows of opposite pairs (Keeper, 2006). M. podia of the family Moonscape was found to exhibit in vitro biostatistics activity (Gents et al. 2008). The seeds and other parts of M. Mammals. However, an unlikely large dose would be necessary to cause such problems in humans. Extracts of the plant have been shown in scientific trials to be a moderate diuretic, depress duodenal contractions similar to atropine colophons, promote regeneration of nerves, and reduce menorahs.
Antidepressant activity has been demonstrated in humans. Root extracts are reported to be a strong emetic, due to the emission (Keeper, 2006). Coleus aromatics is commonly known as “Oregano’ belonging to Lamiae family or Mint family. C. aromatics is used for seasoning meat dishes and in food products, while a detection of its leaves is administered in cases of chronic cough and asthma. It is considered to be an antispasmodic, stimulant and is used for the treatment of headache, fever, epilepsy and dyspepsia.
It is used to treat conditions such as indigestion, diarrhea, nervous tension, insect bites, toothache, earache, rheumatism, whooping cough, and bronchitis. It is also known to be a very powerful painkiller, stimulates flow of bile aiding digestion. Mast cell stability property of C. aromatics leaves were checked in rat peritoneal mast cells. Chinstrap et al. , 2010) A study was previously conducted on the efficacy of oregano oil which contains Carroll and thymus against platonic S. erasure and S. Epidermises, including internationalist strains (Nonstop et al. , 2004).
Plant Extraction Method One of the methods that have been developed for extracting crude plants is detection. Detection is a water-based preparation to extract active compounds of herbs, made by boiling the plant parts for some period. The plant materials are generally into broken into small pieces or grounded into a powder. The smaller they re, the more easily they will be absorbed into the water (Tendon, 2008). General Description of Bacteria of this study S. erasure is a Gram-positive spherical bacteria that occur in microscopic clusters resembling grapes. Taxonomically S. erasure is a member of the bacterial family Staphylococcal.
It colonizes mainly in the nasal passages, but it may be found commonly in most other anatomical locales, including the skin, oral cavity and gastrointestinal tract. S. erasure causes a variety of supportive infections and toxins in humans. S. erasure is unique in its capacity to clot plasma known as coagulate. Coagulate is a traditional marker for identifying S erasure in the clinical microbiology laboratory. Though, there is no overwhelming evidence that it is a virulence factor, although it is reasonable to speculate that the bacteria could protect themselves from phagocyte and immune defenses by causing localized clotting (Attar, 2011).
Abscissas are the laboratory determination of the concentration of a drug or other substance in a specimen by comparing its effect on an organism, an animal, or an isolated tissue with that of a standard preparation (Mossy, 2009). Succulent refers to any of various plants having fleshy leaves or stems that store water (Muffling, 2009). Platonic is a collective name for small animal and plant (phytoplankton) organisms that drift passively in all natural bodies of water (Saunders, 2007).
CHAPTER 2 METHODOLOGY RESEARCH DESIGN This study utilizes experimental research design, specifically two-group design where two comparable groups are employed as experimental and control groups (Calamari and Calamari, 2002). In this experiment, five different herbal plant extracts were compared. All of these of grams of herbal plants and distilled water as solvent. But the concentration of each herbal plant extracts varied namely 50% (1 ml), 75% (1. Ml), 100% (2 ml). Commercially prepared (positive control) was used as the control group known as the positive control set-up of the study.
RESEARCH LOCALE The experiment was conducted at Pharmacy Laboratory 1 (Room 401) of SST. Scholastics College Tactical. For easy access of reagent and laboratory apparatuses, all relevant test and measurements, such as weighing and the extraction of the plants, and performing tube coagulate test were done at Pharmacy Laboratory 1 . However, the inoculation and culturing of S. erasure were done at Medical Technology Laboratory for safe, regulated, and conducive environment. GENERAL PROCEDURES l. Preparation of Herbal Plant Extracts a.
Plant Collection P. Aruba (CALICHES), l. Aquatic (KAYAKING), M. putted (MIKHAIL), E. hart (GATES-GATES), and C. aromatics (OREGANO) samples were collected from the Happy Homes Edit, Tactical City, Letter, Philippines. The individual plant was randomly collected between 8:00 am to 10:00 am by uprooting method. Collected samples were wrapped in clean plastic bags and transported directly to the Pharmacy Laboratory for the preliminary procedures. The samples were thoroughly washed with running water to remove debris.
The plant materials were rinsed with stilled water afterwards then, air dried for two days. Each sample was weighed (500 g). Only the healthy looking matured leaves in every herbal plant were picked and randomly selected to be used for detection. B. Extraction and Purification of Plant Extract Detection of each herbal plant was prepared by boiling egg of the leaves in mall distilled water in a flask for 20 minutes. The flask containing the leaves and detection was removed from the heat and allowed to cool. The content of flask was filtered through filter paper to obtain clear detection.
The aqueous infusion was prepared by aging egg of the leaves in mall distilled water, left for eh at room temperature and filtered to obtain clear Infusion. Millipede filter was used to purify the plant extracts. C. Test for Contaminants in Plant Extract Triplicate Soy Broth (TTS) was used to check the presence of organisms in plant extract. II. Preparation of Bacterial Cultures a. Preparation of Gars Imitation Salt Agar (MS) b. Collection of S. erasure Strains Strains of S. erasure were obtained from an individual with an infected wound associated with pus by using sterile cotton swab.
The sample was immediately treated in Imitation Salt Agar and incubated for 24 hours at 370 C. After 24 hours, growth of creamy, white “butter looking” S. erasure colonies were seen. The Imitation Salt Agar turned pink to yellow due to the ability of S. erasure to ferment imitation in the medium. C. Altercation of Bacteria characteristic classification of Gram positive bacteria, S. erasure. All equipment and supplies needed (Gram stain, microscope slides, heat source, sterile swab, specimen and microscope) are gathered. The slide was labeled with specimen number.
An isolated colony of bacteria of the agar plate was obtained with a sterile swab. The swab was placed on the microscope slide and the colonies were spread in a circular motion. The film was allowed to air dry. The dried film was briefly passed through the Bunsen flame two or three times but the dried film was not exposed directly to the flame. The slide was placed on staining tray above the sink. The surface of the slide was flooded with crystal violet stain and let sit for one minute and washed off briefly with distilled water (not over 5 seconds).
The slide was flooded with gram’s iodine, time for 30 seconds and washed off with distilled water. The slide was flooded with enter stain, seafaring, and let sit for one minute. Then, the slide was rinsed with distilled water. The slide was drained and allowed to air dry. A dark purple crystal violet stain, retained by thick layer of pedagogical which forms the outer layer of the cell, specifies the Gram-positive bacteria. The slide was blotted with a drop of oil and read under oil immersion lens of the microscope and a singly, in pairs, and in clusters of Staphylococci was seen under the microscope.
McFarland Standards was stored in standing position at ICC to ICC and protected from light during 12 weeks. The suspension was whirled again for at least 1 minute to obtain a homogeneous suspension and to break the clumps. Tube Coagulate Test Three tubes were taken and labeled as test (plant extract), positive control, and negative control. Each tube was filled with 0. 5 ml of 1 in 10 diluted human plasma. Readily prepared human plasma was bought at Red Cross to have standard source of plasma. 0. 1 ml of overnight broth culture of test bacteria S. erasure was added each of every three tubes.
All the tubes were incubated at CHIC and observed up to four hours. Positive result was indicated by gelling of the plasma, which remains in place even after inverting the tube. The tests were scored as (4 +) the fibrin clot fills the omelet volume occupied by the broth; (3 +) the clot fills more than half but less than the total volume occupied by the broth; (2 +) the clot fills less than half the total volume occupied by the broth; (1 +) there is a little disorganized clot formation; (negative) no clot observed but a little amorphous deposit might be seen.
After 24 hours some clot structure was lost, and this was recorded as a shrinkage. This may have been due to disturbance of the tubes in taking earlier readings or to the activity of bacterial proteases. All of the tests were carried out in duplicate. Al. Antibacterial Assay of Herbal Plant Extract