Mango (Mangifera Indica) Leaves Extract and Coconut Oil as an Antibacterial Ointment

Mango (Mangifera indica) leaves extract and Coconut Oil as an Antibacterial Ointment A Science Investigatory Project of Kristifany C. Mamba Bansud National High School-Regional Science High School for Region IV – MIMAROPA Pag-asa, Bansud, Oriental Mindoro Abstract The purpose of this study is to produce an antibacterial ointment out of Mango leaves extract and coconut oil. Young mango leaves were gathered and was chopped into small pieces. 50 mL of coconut oil was put in a frying pan. Then, the chopped mango leaves was added to the coconut oil. It was mixed for 10 minutes. Next, the small bits of mango leaves were removed from the coconut oil. Lastly, candle wax was added to the mixture. It was stirred again thoroughly. The solution was transferred into an empty container and left to cool down. The mango leaves extract with coconut oil was tested at the Bureau of Plant Industry. It was tested against the bacteria E. coli and S. aureus. The resulting Numerical value was 2. 5 for E. coli and 3. 0 for S. aureus. The bureau used the standard parameter 1-2- slightly inhibited and 3-5 as partially inhibited. Thus, the inhibition of the mango leaves extract and coconut oil in E. oli was slight and it is partial in S. aureus. The researcher concluded that mango leaves extract with coconut oil can kill bacteria like E. coli and S. aureus. It can also be concluded that it can help wounds heal faster because of its antibacterial property. Chapter I Introduction Background of the Study The Philippines have many different herbal plants that can cure different illne ss like body pain, toothache, arthritis, and other diseases. The herbal plants we have contain helpful constituents and properties that can cure different kinds of diseases. We can make useful product made from these plant and other materials. Nowadays, many herbal plants are being discovered with more uses. Many companies used herbal plants to make ointments, tablets, coffee or teas. Ointments are very useful in treating different kind of wounds. The production of ointments from herbal plants found in our country can help us minimize our dependence on the use of high – cost ointments. The herbal plant must have anti-inflammatory, anti-allergenic and antibacterial properties to produce an effective ointment. Most of the wounds are infected by the common bacteria like Escherichia coli. As the wounds go deeper and become more complex they can infect the underlying muscles and bone causing osteomyelitis. Coliforms and anaerobes are associated with osteomyelitis in those people who have infected wounds. You also see the bacteria Staphylococcus aureus in the infected wound. Local factors that increase chances of wound infection are having large wound area, increased wound depth, degree of chronicity, the body, necrotic tissue, and mechanism of injury (bites, perforated viscus). (Neal R. Chamberlain. n. . ) The mango leaves (Mangifera indica) and coconut oil possesses antibacterial activity against different bacteria. (Research Update of Mango and Mango Leaf Extract, n. d. ). Coconut and olive oils are traditionally used to moisturize and treat skin infections. Extensive research done by scientists such as Jon J Kabara, PhD, has shown that the Lauric acid found in Coconut Oil is a potent antim icrobial agent  . Lauric acid is a major component (49%) of Coconut oil. It has also been found to kill the H. Pylori bacteria  in the stomach which are responsible for many stomach problems such as ulcers. The good thing about Lauric acid is that it doesn't kill friendly bacteria in the stomach. Antibiotics kill both good and bad bacteria in the stomach and often need to be followed with probiotics such as acidophilus bacteria to replenish friendly bacteria in the gut. Objectives General Mangifera indica leaves and coconut oil have anti-bacterial contents which can help remove the infection on the wounds. This study aimed to produce an ointment which can kill the bacteria and cure different types of wounds out of Mangifera indica leaves and coconut oil. Specific This research study was conducted to determine if mango leaves extract and coconut oil can be made into an ointment and if it can help wound heal faster. Statement of the Problem Specifically, this study ought to answer the following questions: 1. Can the ointment made from Mangifera indica leaves extracts and coconut oil kill the bacteria in the wounds? 2. Can the Mangifera indica leaves extract and coconut oil be made into an ointment? 3. Can the ointment made from Mangifera indica leaves and coconut oil extract help the wound to heal faster? Hypothesis 1. The ointment made from Mangifera indica leaves extracts and coconut oil can kill bacteria in the wounds. 2. The extract of Mangifera indica leaves and coconut oil can be made into an ointment for curing wounds. Significance of the Study This study greatly benefits the people in the community who cannot afford to buy expensive ointment for wounds. It can also benefit the hospitals and in small clinics. The additional medication in curing wounds can help a lot to save a life. It has significance to those who were far from the store or drug store because they can cure our wounds without taking too long from buying ointments from far drugstores. It can be also a source of income for the people in provinces. Scope and Limitation This study was limited only on the production of ointments from mango leaves extracts and coconut oil. The ointment produced from mango leaves extracts and coconut oil focuses on killing the bacteria in the wounds. It was limited to use if there is irritation on the skin after the application of the ointment. For the patients who have sensitive skin should ask permission from a doctor before using the ointment. Chapter II Review of Related Literature Review of Related Literature Antibacterial Pertaining to a substance that kills bacteria or inhibits their growth or replication. Antibiotics synthesized chemically or derived from various microorganisms exert their bactericidal or bacteriostatic effect by interfering with the production of the bacterial plasma wall; by interfering with protein synthesis, nucleic acid synthesis, or plasma membrane integrity; or by inhibiting critical biosynthetic pathways in the bacteria. (2009, Elsevier. ) E. coli E. coli  is a common type of  bacteria  that can get into food, like beef and vegetables. E. oli  is short for the medical termEscherichia coli. E. coli  normally lives inside your intestines, where it helps your body break down and digest the food you eat. Unfortunately, certain types (called strains) ofE. coli  can get from the intestines into the blood. This is a rare illness, but it can cause a very serious infection. (Steven Dowshen, MD, August 2009) S. aureus Staphylococci (staph) are Gram-positive s pherical 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  Staphylococcus albus  (white). The latter species is now named  Staphylococcus epidermidis. Although more than 20 species of  Staphylococcus  are described in Bergey's Manual (2001), only  Staphylococcus aureus  and  Staphylococcus epidermidis  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. S. ureus  is often hemolytic on blood agar;  S. epidermidis  is non hemolytic. The bacteria are catalase-positive and oxidase-negative. S. aureus  can grow at a temperature range of 15 to 45 degrees and at NaCl concentrations as high as 15 percent. Nearly all strains of  S. aureus  produce the enzyme coagulase: nearly all strains of  S. epidermidis  lack this enzyme. S. aureus  should always be considered a potential pathogen; most strains of  S. epidermidis  are nonpathogenic and may even play a protective role in humans as normal flora. Staphylococcus epidermidis  may be a pathogen in the hospital environment. Pathogenesis of  S. aureus  infections Staphylococcus aureus  causes a variety of suppurative (pus-forming) infections and toxinoses in humans. It causes superficial skin lesions such as  boils,  styesand  furuncules; more serious infections such as  pneumonia,   mastitis, phlebitis,  meningitis, and  urinary tract infections; and deep-seated infections, such as  osteomyelitis  and  endocarditis. S. aureus  is a major cause of  hospital acquired (nosocomial) infection  of surgical wounds and infections associated with indwelling medical devices. S. aureus  causes  food poisoning  by releasing enterotoxins into food, and  toxic shock syndrome  by release of superantigens into the blood stream. S. ureus  expresses many potential  virulence factors: (1)  surface proteins that promote colonization of host tissues; (2) invasins that promote bacterial spread in tissues (leukocidin,  kinases,  hyaluronidase); (3) surface factors that inhibit phagocytic engulfment (capsule,  Prote in A); (4) biochemical properties that enhance their survival in phagocytes (carotenoids,  catalase production); (5) immunological disguises (Protein A,  coagulase); (6) membrane-damaging toxins that lyse eucaryotic cell membranes (hemolysins, leukotoxin, leukocidin; (7) exotoxins that damage host tissues or otherwise provoke symptoms of disease (SEA-G,  TSST,  ET); and (8) inherent and acquired resistance to antimicrobial agents. Membrane-damaging toxins alpha toxin (alpha-hemolysin)  The best characterized and most potent membrane-damaging toxin of  S. aureus  is alpha toxin. It is expressed as a monomer that binds to the membrane of susceptible cells. Subunits then oligomerize to form heptameric rings with a central pore through which cellular contents leak. In humans, platelets and monocytes are particularly sensitive to alpha toxin. Susceptible cells have a specific receptor for alpha toxin which allows the toxin to bind causing small pores through which monovalent cations can pass. The mode of action of alpha hemolysin is likely by osmotic lysis. ?-toxin  is a sphingomyelinase which damages membranes rich in this lipid. The classical test for ? -toxin is lysis of sheep erythrocytes. The majority of human isolates of  S. aureus  do not express ? -toxin. A lysogenic bacteriophage is known to encode the toxin. (2008 Kenneth Todar, PhD  ) delta toxin  is a very small peptide toxin produced by most strains of  S. aureus. It is also produced by  S. epidermidis. The role of delta toxin in disease is unknown. Leukocidin  is a multicomponent protein toxin produced as separate components which act together to damage membranes. Leukocidin forms a hetero-oligomeric transmembrane pore composed of four LukF and four LukS subunits, thereby forming an octameric pore in the affected membrane. Leukocidin is hemolytic, but less so than alpha hemolysin. Only 2% of all of  S. aureus  isolates express leukocidin, but nearly 90% of the strains isolated from severe dermonecrotic lesions express this toxin, which suggests that it is an important factor in necrotizing skin infections. (2008 Kenneth Todar, PhD  ) Wound Healing Wound healing is a complex process with many potential factors that can delay healing. There is increasing interest in the effects of bacteria on the processes of wound healing. All chronic wounds are colonized by bacteria, with low levels of bacteria being beneficial to the wound healing process. Wound infection is detrimental to wound healing, but the diagnosis and management of wound infection is controversial, and varies between clinicians. There is increasing recognition of the concept of critical colonization or local infection, when wound healing may be delayed in the absence of the typical clinical features of infection. The progression from ound colonization to infection depends not only on the bacterial count or the species present, but also on the host immune response, the number of different species present, the virulence of the organisms and synergistic interactions between the different species. There is increasing evidence that bacteria within chronic wounds live withi n biofilm communities, in which the bacteria are protected from host defences and develop resistance to antibiotic treatment. (Edwards R,  Harding KG Apr. 17, 2004) Bacteria and Wounds Bacteria are ubiquitous in the geography of the human body. In the skin, the average human being harbors at least 200 species of bacteria, totaling more than 1012 organisms. Therefore, when the skin is broken by trauma or disease, bacteria are also ubiquitous in wounds. When discussing the presence of bacteria in an open wound of a human host, three conditions are noted with respect to their presence on or in the tissue, their impact on the healing of the wound, and the associated immune response from the host. The first condition is bacterial contamination or the simple existence of bacteria on the surface of the wound. Contamination is specifically defined as the presence of non-proliferating organisms on the superficial tissues. Contaminating bacteria do not elicit an immune response from the hos t and do not impact the healing process. The second condition, bacterial colonization, is differentiated from contamination in that it refers to proliferating organisms on the wound surface – bacteria that have adhered to the superficial tissues and have begun to form colonies. Colonization is also characterized by a lack of immune response from the host and generally is not believed to impact or interfere with the healing process. 2 Wounds that contain nonviable tissue (ie, slough and/or eschar) offer a particularly hospitable environment for colonization because the dead tissues provide a ready source of nutrients for the growing bacterial colonies. In the third condition, bacterial infection, proliferating bacteria are not only present on the surface of the wound or in nonviable tissue, but have also invaded healthy, viable tissue to such a depth and extent that they elicit an immune response from the host. Local clinical signs of tissue redness, pain, heat, and swelling generally characterize this immune response, along with an increase in exudate production or purulence. Bacterial infection delays and may even halt the healing process. The mechanism of this healing delay involves competition between host cells and bacterial cells for oxygen and nutrients and increased host cell production of inflammatory cytokines and proteases in response to the bacteria and their associated toxins. (Liza Ovington, PhD, CWS, n. d) Related studies In the research update of mango and mango leaf extract, effects of a natural extract from Mangifera indica L, and its active compound, mangiferin, on energy state and lipid peroxidation of red blood cells. Following oxidative stress, modifications of several biologically important macromolecules have been demonstrated. In this study they investigated the effect of a natural extract from Mangifera indica L (Vimang), its main ingredient mangiferin and epigallocatechin gallate (EGCG) on energy metabolism, energy state and malondialdehyde (MDA) production in a red blood cell system. Analysis of MDA, high energy phosphates and ascorbate was carried out by high performance liquid chromatography (HPLC). Under the experimental conditions, concentrations of MDA and ATP catabolites were affected in a dose-dependent way by H(2)O(2). Incubation with Vimang (0. , 1, 10, 50 and 100 mug/mL), mangiferin (1, 10, 100 mug/mL) and EGCG (0. 01, 0. 1, 1, 10 muM) significantly enhances erythrocyte resistance to H(2)O(2)-induced reactive oxygen species production. In particular, they demonstrate the protective activity of these compounds on ATP, GTP and total nucleotides (NT) depletion after H(2)O(2)-induced damage and a reduction of NAD and ADP, which both increase because of the energy consumption following H(2)O(2) addition. Energy charge potential, decreased in H(2)O(2)-treated erythrocytes, was also restored in a dose-dependent way by these substances. Their protective effects might be related to the strong free radical scavenging ability described for polyphenols. Mango and Mango Leaf Extract, n. d. ) Mangifera indica L. extract consists of a defined mixture of components (polyphenols, terpenoids, steroids, fatty acids and microelements). It contains a variety of polyphenols, phenolic esters, flavan-3-ols and a xanthone (mangiferin), as main component. This extract has antioxidant action, antitumor and immunemodulatory effects proved in experimental models in both in vitro and in vivo assays. The present study was performed to investigate the genotoxicity potential activity of Vimang assessed through different tests: Ames, Comet and micronucleus assays. Positive and negative contr ols were included in each experimental series. Histidine requiring mutants of Salmonella typhimurium TA1535, TA1537, TA1538, TA98, TA100 and TA102 strains for point-mutation tests and in vitro micronucleus assay in primary human lymphocytes with and without metabolic activation were performed. Results of Comet assay show that the extract did not induce single strand breaks or alkali-labile sites on blood peripheral lymphocytes of treated animals compared with controls. On the other hand, the results of the micronucleus studies (in vitro and in vivo) show Vimang induces cytotoxic activity, determined as cell viability or PCE/NCE ratio, but neither increased the frequency of micronucleated binucleate cells in culture of human lymphocytes nor in mice bone marrow cells under their experimental conditions. The positive control chemicals included in each experiment induced the expected changes. The present results indicate that M. indica L. extract show evidences of light cytotoxic activity but did not induce a mutagenic or genotoxic effects in the battery of assays used. (Mango and Mango Leaf Extract, n. d. ) Anti-allergic properties of Mangifera indica L. extract (Vimang) and contribution of its glucosylxanthone mangiferin. : Vimang is the brand name of formulations containing an extract of Mangifera indica L. , ethnopharmacologically used in Cuba for the treatment of some immunopathological disorders, including bronchial asthma, atopic dermatitis and other allergic diseases. However, the effects of Vimang on allergic response have not been reported until now. In this study, the effects of Vimang and mangiferin, a C-glucosylxanthone isolated from the extract, on different parameters of allergic response are reported. Vimang and mangiferin show a significant dose-dependent inhibition of IgE production in mice and anaphylaxis reaction in rats, histamine-induced vascular permeability and the histamine release induced by compound 48/80 from rat mast cells, and of lymphocyte proliferative response as evidence of the reduction of the amount of B and T lymphocytes able to contribute to allergic response. In these experiments, ketotifen, promethazine and isodium cromoglicate were used as reference drugs. Furthermore, they demonstrated that Vimang had an effect on an in-vivo model of inflammatory allergy mediated by mast cells. These results constitute the first report of the anti-allergic properties of Vimang on allergic models, as well as suggesting that this na tural extract could be successfully used in the treatment of allergic disorders. Mangiferin, the major compound of Vimang, contributes to the anti-allergic effects of the extract. (Mango and Mango Leaf Extract, n. d. ) Anti-inflammatory, analgesic and hypoglycemic effects of Mangifera indica Linn. (Anacardiaceae) stem-bark aqueous extract.   Previous studies in their laboratories and elsewhere have shown that some members of Anacardiaceae family possess antiinflammatory, analgesic and hypoglycemic effects in man and mammalian experimental animals. The present study was, therefore, undertaken to examine the antiinflammatory, analgesic and antidiabetic properties of the stem-bark aqueous extract of Mangifera indica Linn. , M. indica a member of the Anacardiaceae family, in rats and mice. The stem-bark powder of M. indica was Soxhlet extracted with distilled water and used. M. indica stem-bark aqueous extract (MIE, 50-800 mg/kg i. p. ) produced dose-dependent and significant (p