PHYTOCHEMICAL SCREENING AND ANTIMICROBIAL ACTIVITIES OF AQUEOUS AND METHANOLIC BUD EXTRACT OF Syzigium aromaticum (CLOVE) ON SOME SELECTED PATHOGENIC MICROORGANISMS

PHYTOCHEMICAL SCREENING AND ANTIMICROBIAL ACTIVITIES OF AQUEOUS AND METHANOLIC BUD EXTRACT OF Syzigium aromaticum (CLOVE) ON SOME SELECTED PATHOGENIC MICROORGANISMS

ABSTRACT

This study is aimed at analyzing the phytochemicals and elucidating the antimicrobial properties of Syzyigium aromaticum (clove) which is one of the most popular and widely used spice that has been prepared to possess high nutritional value, antimicrobial properties. 100% aqueous and methanol (water and alcohol) for Methanolic and aqueous extracts were prepared. About 6 phytochemical compounds were detected qualitatively in all of the two different extracts which include Tannins, Saponins, Alkaloids, Terpenoids, Cardiac glycosides and flavonoid. Agar well diffusion method was used to test the anti bacterial and anti fungi properties of Syzyigium aromaticum (clove) was found that all pathogenic bacteria and fungus were inhibited by the clove. The Phytochemical Screening of Aqueous and Methanolic bud extract of Syzigium aromaticum (clove) was carried out to determine the presence of secondary metabolites in the Aqueous and Methanolic extract of Syzigium aromaticum (clove) bud. The result of phytochemical screening shows that there is presence of Tannis, Saponins, Alkaloid, Terpenoid in both aqueous and Methanolic extract while there is absence of cardiac glycoside in both aqueous and Methanolic extract. There is absence of flavanoid in methanol while in aqueous is present. The Antimicrobial activity of Syzigium aromaticum Aqueous and Methanolic bud extract was determined by agar well diffusion method on some selected pathogenic microorganisms (Streptococcus spp, Klebsialla spp, Aspergillus niger and Candida albicans) at 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml concentrations. The result of the antimicrobial activity of aqueous and Methanolic extract of Syzigium aromaticum (clove) bud shows that Methanolic and aqueous extract is effective on some selected pathogenic microorganism. The zone of inhibition of Methanolic extract against Aspergillus was 15.85mm, Candida 13.33mm, Klebsiella 19.88mm, Streptococcus 20.75mm at 200mg/ml while the zone of inhibition of Aqueous at 200mg/ml was Aspergillus 10.61mm, Candida 10.88mm, Klebisiella 18.02mm, Streptococcus 20.75mm. The Aqueous and Methanolic extract of Syzigium aromaticum (clove) bud showed Minimum Inhibitory Concentration (MIC) by broth dilution method. The lowest concentration of the aqueous and metanolic extract that inhibited the growth of fungi and bacteria showed at concentration of 100mg/ml. The growth of Aspergillus and Candida. At 50mg/ml Methanolic extract inhibited growth of Aspergillus and Candida while at 50mg/ml of the extract concentration of aqueous and methanol, the growth of Streptococcus and Klebsiella species were both inhibited. Where as, the Minimum Microbiocidal Concentration of aqueous and Methanolic extract of Streptococcus showed 200mg/ml of aqueous extract kills the Streptococcus species, while at 100mg/ml of methanolic extract kills Streptococcus species. In same manner at 100mg/ml of aqueous and methanolic extract, Klebsiella species was killed.

CHAPTER ONE

1.0                                                             INTRODUCTION

1.1       Background of the study

Plant products have a long history of usage in the treatment of a wide range of ailments in many regions of the world, including Nigeria. Many different plant items such as leaves, stem, roots, flower, bark etc have been used in traditional medicine for thousands of years for a variety of purposes because of their antibacterial, anti-inflammatory, cytostatic, antioxidant, anti -fungal, and antiviral properties, many herbal medicines have been utilised for a variety of ailments (Salvamani, et al., 2014).

The World Health Organization defined a medicinal plant as any herbal preparation produced by subjecting plant materials to extraction, fractionation, purification, concentration or other physical or biological process which may be produced for immediate consumption or as a basis for herbal products (WHO, 2021). Phytochemicals often referred to as “secondary metabolities” chemical compounds formed during the plant normal metabolic processes, they were first described at the beginning of the 19th century. The most important of these bioactive compounds of plants are Alkaloids, Flavanoids, Quinones, Phenolic Compounds, Saponins, Tannins, Coumarins, Glycosides, Gums, Polysaccharides, terpenes and other chemical compounds (Sen and Chakraborty, 2016).

Phytochemicals are bioactives and function to protect plants against invasion, disease, and infection. Phytochemical screening is very important in identifying new sources of therapeutically and industrially important compounds like alkaloids, flavonoids, phenolic compounds, tannins etc. In India, phytochemicals, as well as medicinal plants, have remained the most abundant source of healthcare and life improvement since very long (Mujeeb et al., 2014). Ayurvedic, Unani and Siddha medico-therapeutics have played a very important role in Indian society since ancient times. Now in the modern era, about 24%-27% drugs are derived from plant sources. India has one of the richest plant biodiversity in the world especially with Western Ghats and Himalayas being regions rich in plant biodiversity within the country (Mujeeb, et al., 2014).

Antimicrobial activity is the ability of a substance to inhibit or kill bacterial cells (Reiner, 2014). The effectiveness of currently available antibiotics is decreasing due to the increasing number of resistant strains causing infections. The multidrug resistant strains of many microorganisms have revealed exploration of alternative antimicrobial agent (Sharmeen, et al., 2012). The antimicrobial properties of plants have been investigated by a number of researchers worldwide, especially in Latin America. In Argentina, a research tested 122 known plant species used for therapeutic treatments. It was documented that among the compounds extracted from these plants, twelve inhibited the growth of Staphylococus aureue, ten inhibited Escherichia coli, and four inhibited Aspergillus niger and also reported that the most potent compound was extracted from Tabebuia impetiginosa (Sharmeen, et al., 2012)

Clove has been used extensively in traditional medicine for more than 2,000 years in both Indian and Chinese cultures. Because of their capacity to preserve foods and hide the scent of poorly- kept foods, cloves were extremely popular as a therapeutic flower in Europe during the fourth and eighth centuries A.D. Arabic traders introduced the buds to the continent. Clove belongs to the family of Myrtaceae and the genus Syzigium of and species name Syzigium aromaticum. Internationally the name is varied for example Laung, Laumg or Lavang in Hindi, Kabsh qarunfil or kabsh qaranful in Arabic, Ding xiang in Chinese, Clavos de olor in Spanish, Clous de girofle and Kaninfari in Nigeria (Hausa) (Nascimento, et al., 2020).

 Clove has a variety of chemical components, the most important of which are sesquiterpenes volatile oil (eugenol), caryophyllene, tannins, and gum. It has been demonstrated to be a powerful chemo-preventive agent, and ithas been utilised by traditional Ayurvedic physicians in India to treat respiratory and intestinal problems since ancient times. Among the essential oils found in clove flower buds are acetate, eugenol, caryophyllene, alpha-terpinyl methyl eugenol, aipha-humulene, eugenyl, actyl eugenol, .chavicol, naphthalene, methyl salicylate pinene, heptanone, vanillin, sesquiterpenes, and naphthal (Nascimento et al, 2020). Because of its various applications, this spice must be widely planted in most of the places where climatic conditions are conducive to its optimal growth and development. In this way, a maximum yield of its many useable pieces might be reached, allowing for the production of the greatest possible number of commodities of varying natures for the benefit of mankind (Dashti and Morshedi, 2019). Therefore, taking into consideration all of the information, it is necessary to establish its phytochemical constituents, antimicrobial activities and other diverse qualities that are beneficial to humans. The current research will sought to examine the photochemical screening and antimicrobial activities of Syzigium aromaticurn (Clove) aqueous and methanolic extract on some selected pathogenic microorganisms.

1.2       Statement of the problems

Even though pharmacological industries have produced a number of new antibiotics in the last three decades, resistance to these drugs by microorganisms has increased. In general, bacteria have the genetic ability to transmit and acquire resistance to drugs, which are utilized as therapeutic agents. Such a fact is cause for concern, because of the number of patients in hospitals who have suppressed immunity, and due to new bacterial strains, which are multi- resistant. Consequently, new infections can occur in hospitals resulting in high mortality.
The problem of microbial resistance is growing and the outlook for the use of antimicrobial drugs in the future is still uncertain, Therefore, actions must be taken to reduce this problem. for example, to control the use of antibiotic, develop research to better understand the genetic mechanisms of resistance, and to continue studies to develop new drugs, either synthetic or natural. The ultimate goal is to offer appropriate and efficient antimicrobial drugs to the patient.  For a long period of time, plants have been a valuable source of natural products for maintaining human health, especially in the last decade, with more intensive studies for natural therapies.

1.3       Justification of the Problem

Syzygium aromaticurn has attracted the attention due to the potent anti-oxidant and antimicrobial activities standing out among the other spices of plant source. It is one of the most valuable plant that have been used as food preservative and for many medicinal purposes. Syzygium aromaticum is an important medicinal plant due to the wide range of pharmacological effects consolidated from traditional use for centuries. The antimicrobial activities of clove have been proved against several bacterial and fungal strains. Sofia et al, (2007) tested the antimicrobial activity of different Indian spice plants as mint, cinnamon, mustard, ginger, garlic and clove, the only sampled that showed complete bactericidal effect against all the food-borne pathogens tested Escherichia coli, Staphylococcus aureus and Bacillus cereus was the aqueous extract of Syzygium aromaticum at 3% (Sofia, et al., 2007). Going by the enormous importance of Syzygium aromaticum, it has become imperative to carry out a phytochemical screening and antimicrobial activities of Syzigium aromaticurn bud (clove) aqueous and methanolic extract on some selected pathogenic microorganisms.

1.4       Aim of the study

The aim of this study is to examine the phytochemicals properties and antimicrobial activities of Syzigium aromaticum bud (clove) aqueous and methanolic extract on some selected pathogenic microorganism

1.5       Objectives of the Study

i.To carry out phytochemical screening on clove aqueous and methanolic extract

 ii.To determine the antimicrobial effects of aqueous and methanolic extract of clove bud on some selected pathogenic microorganism

 iii. To determine the Minimum Inhibitory Concentration (MIC) and Minimum Microbicidal Concentration (MMC).