Abstract

This project aimed at analyzing the antimicrobial activity of natural honey and ginger harvested from farms within the Buea community on the normal flora of the skin (Staphylococcus aureus). Actually, there are many microorganisms on the skin surface of bacteria origin acting as normal flora. Staphylococcus aureus occupy 60% to 80% of the total number of microorganism found on the skin of bacterial origin. Secondly, it is one of the most resistant of all none spore forming bacteria. Thirdly, most strains of Staphylococcus aureus are heat stable withstanding temperatures as high as 500c for 30mins and more so can resist a high concentration of salt (7.5% to 9%), sodium chloride, and it causes skin infection, infect sebaceous glands, invade damaged abrasions and cuts causing boils, furuncle. Honey and ginger were bought from the local farmers, selling their produce within the Buea central markets and were transported to the life science laboratory of the University of Buea. Ginger bulbs were cut, washed, and rinsed several times under hygienic conditions and then crushed using a grater to get the ginger extract. Susceptibility discs were prepared from sterile filter papers using a disc borer in which the ginger extract and honey were impregnated. Skin swab specimens were collected from different body parts using nine sterile swab sticks and inoculated on Mannitol salt agar (MSA) and later subcultured on nutrient agar (NA), after which isolated colonies were characterized by gram staining and Biochemical tests. Colonies were subculture on Mueller Hinton agar (MHA) for susceptibility testing. After 24hr of incubation of the skin sample on mannitol salt agar 3/5 of the colonies appeared yellow, with the media appearing yellow for most of the plates, indicating Staphylococcus aureus subculture of the mannitol positive colonies on nutrient agar resulted in cream white colonies and non-uniform growth. Gram stain of the colonies from the nutrient agar plate revealed gram positive cocci. The colonies were both coagulase positive and catalase positive. The isolated Staphylococcus aureus was susceptible to both honey and ginger. Yellow colour on the mannitol salt agar plate results from the breakdown of mannitol by the bacteria to releasing acid which changes the colour of the dye to yellow. The appearance of purple round cells in irregular clusters was a clear indication of gram positive, giving up of active bubbles wen the colonies were mixed with hydrogen peroxide(H2O2), was also an indication that the bacteria isolate was staphylococcus aureus although it is not the only catalase positive staphylococcus species. For the coagulase, clumping occurred few seconds after the colonies came in contact with plasma, thus confirming that the bacteria isolate was Staphylococcus aureus. Zones of inhibition around the susceptibility discs was an indication that honey and ginger had antimicrobial activity on staphylococcus aureus. Susceptibility varied with the dilution of the honey and ginger with a greater inhibition around the disc impregnated with the stock, thus honey and ginger can be used for the treatment of infections caused by staphylococcus aureus.

CHAPTER ONE

INTRODUCTION AND LITERATURE REVIEW

1.1 INTRODUCTION

Antimicrobial agents are substances known to have therapeutic effect on microorganisms either as a control prevention or cure of microbial and non-microbial disease origin. These antimicrobial agents are synthesized chemotherapeutic substances obtained majorly from microorganisms, plants and some animal products. The failure of these antibiotics has caused men to search for more effective sources of natural products from plants and some insects. Though some of these products perform less or higher than synthesized antibiotics, in some cases, they have been found safe and good source of pharmacological effect for man. Medicinal values are derived not only from the already available drugs in man health care system but by invention most especially into plants for the derivation properties which are medicinal.

Products derived from plants have been used for medicinal purposes for centuries; at present about 80% of the world population relies on botanical preparation as medicines to meet their health needs. Many scientists have reported plants extracts. Studies have shown that 60% to 80% of the body’s micro flora is compose of Staphylococcus aureus which is a member of the firmicutes. Staphylococcus aureus is a species from the genus Staphylococci. Species also found include; Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus felis and Staphylococcus arlettae. Of all the species, Staphylococcus aureus is found to be the most populated and is a common cause of skin infection including abscesses, respiratory infection such as sinusitis, infect wound. It is mostly found in high amounts in openings of the body and cracks such as wounds on the skin surface where they easily get into the body and cause infections. Pathogenic strains often promote infections by producing virulent factors such as potent protein toxins. It is one of the most common cause of hospital acquired wound infections following surgery.

1.2 – LITERATURE REVIEW

1.2.1 Antibacterial activities of honey

Honey is a sweet viscous food substance produced by Apismellifera, commonly called bee, and some related insects. Bees produced honey from sugary secretions from plants and other insects through regurgitation, enzymatic activity and water evaporation. It contains some special type of substances that have antimicrobial properties, meaning they kill or inhibit the growth of microorganisms like bacteria, viruses or fungi. Honey has a low pH, which is not favorable for the growth of most bacteria, also, it has a high sugar content, making it hygroscopic which draws water from the environment and dehydrate the bacteria cell,

Furthermore, honey has hydrogen peroxide produced by bee derived enzyme glucose oxidase (activity more stable at 40C than at 250C). These properties give honey a broad spectrum antibacterial effect to treat serious medical conditions caused by bacteria such as Staphylococcus aureus and so on. Infections such as wound infections, pneumonia and other bacterial skin infections.

1.2.2 Antibacterial activities of ginger

Zingiber officinale is a flowering plant whose bulb (Ginger root) or simply ginger is widely used as a spice or a folk medicine. Ginger like honey also has some antibacterial substances which help to kill or inhibit the growth of microorganisms like bacteria, fungi etc. Ginger has been valued for it antibacterial activities for thousands of years in Asian culture(Weil, 2005). Its higher inhibition is due to its ethanol and methanol content found in its extract. It was noticed that increasing the concentration of the extract leads to more inhibitory effect on the growth of bacteria. The methanolic extract posse phenolics, saponin, tannin, and flavonoids which contribute to its antibacterial activity in one way or the other. On the other hand, cardiac glycoside is found in the aqueous extract of ginger. These substances gives ginger a broad spectrum of activity, to treat respiratory infections caused by bacteria and viruses as well as other skin or wound infections caused by Staphylococcus aureus and other bacteria.

1.2.2.1 Effect of honey and ginger on normal skin flora

Normal micro flora is found on the surface of all human skin (Prescott et al, 2008). The skin is an important organ of the body that serves for protection against infections by germ and shields delicate underlying tissue against injury (Speer and Dawnson, 1965). If an individual loses this protection either by injury or by surgical operations, the person is much more susceptible to infection on the skin. Most of the micro flora found on the human skin are harmless and some are beneficial. It is of great importance to note that every microorganism is a potential pathogen because; even the most harmless organism may cause infection particularly if the skin is broken. Some of the normal micro flora of the skin that are of bacterial origin include; Mycobacterium spp, Candida albicans, Staphylococcus epidermidis, Mallasseiza furfur, Staphylococcus aureus. Resident microbes can cause skin disease and enter the blood stream creating life threatening disease particularly in immunosuppressed people (Alam et al, 1990). Normal flora may serve as defense against colonization by potential pathogens, for instance treatment of the skin of humans with antibacterial agent such as hexachlorophene results in suppression of the normal gram-positive flora and promote colonization and clinical infection by gram negative bacilli and other organisms that cannot normally establish themselves on the skin (Thuma et al, 2013).

Honey has a broad spectrum antibacterial activity. Honey has seen a revival recently in the Western medical field, as it has shown inhibitory activity against a range of detrimental and antibiotic-resistant microbes of infected wounds.

1. Honey may be the first recorded medicine having been documented in the Smith Papyrus of Egypt, which dates to between 2200-2600 BC
2. Since ancient times honey has been renowned for it wound- healing properties.
3. The overwhelming use of antibiotics has resulted in widespread resistance, therefore alternative antimicrobial strategies are necessary
4. Honey has demonstrated potent in vitro activity against antibiotic-resistant bacteria and it has been successfully applied as treatment of chronic wound infections not responding to antibiotic therapy 3. For example, honey has received attention as an important tool against strains such as Methicillin-resistant Staphylococcus aureus.
5. No microbial resistance against honey has been observed, making it attractive as a treatment for wound infections.
6. Honey possesses several antimicrobial properties and can act through various mechanisms of action. The antimicrobial potency and medical applications of honey are tremendous as it has demonstrated inhibitory effects against a number of pathogenic bacteria.

Ginger has a broad-spectrum antibacterial effect. Interest in plants with antibacterial effect has been revived as a result of microbial resistance on prescribed drugs. The resistance could be indiscriminate use of commercial drugs or not taking an antibiotic prescription according to the instruction, for example not taking all the prescription for the treatment of infectious diseases (Aliero and Afolayan, 2006). Ginger has been found effective for the treatment of human ill-health like respiratory tract infections like pneumonia, sore throat caused by bacteria and other systemic diseases such as erectile difficulties, kidney stones, Reynaurd’s disease and viral infections (Peggy 2006) and even fungal infections such as athlete’s foot. Ginger contain several antimicrobial properties and these properties have different mechanisms of action.

Ginger is also effective against toxins deposited in food by bacteria such as Staphylococcus aureus in food poisoning, more so, it has inhibitory effect on a number of pathogenic bacteria.

1 2.3 Staphylococcus aureus

1.4 STATEMENT OF THE PROBLEM

Honey and ginger are used in the Buea community to treat respiratory as well as skin infections. Honey and ginger are commonly used in the area, however, it is not known between the honey and ginger which one of them has a stronger antibacterial effect as those who use them to treat bacterial infections claim that they have antibacterial properties. Therefore, it is of great importance to Annalise the antibacterial strength of honey and ginger as more people increasingly make more use of them. This information will also help the population to know which one of them will be preferable to use in case of any bacterial skin infection.

1.5 RATIONALE

To analyze the antibacterial strength of honey and ginger commonly used within the Buea community to treat respiratory, skin as well as other infections caused by bacteria such as Staphylococcus aureus is of great significance. The availability of the little information on the antimicrobial strength of natural honey and ginger harvested from farms around the Buea community will be good. Therefore, analyzing the antibacterial strength of honey and ginger harvested from the farms within the Buea community will provide some information to the population on which of them can effectively be used to treat skin or respiratory infections caused by bacteria such as Staphylococcus aureus.

1.6 HYPOTHESIS

Natural honey and ginger extracts harvested from the farm have antibacterial effect on Staphylococcus aureus and can be used to treat infections caused by Staphylococcus aureus when used.

1.7 OBJECTIVES

1.7.1 Main objective

To analyze the antibacterial strength of natural honey and ginger harvested from the farm on Staphylococcus aureus.

1.7.2 Specific objective

1. To isolate Staphylococcus aureus from other skin micro flora
2. To analyze antimicrobial activity of the natural honey and ginger using the isolated Staphylococcus aureus.