Our Blog

List of recently published project topics and materials

PROJECT TOPIC- ANTIMICROBIAL ACTIVITY OF METHANOL EXTRACT AND FRACTIONS OF MORINGA OLEIFERA LAM. ROOT BARK ON CLINICAL ISOLATES OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS

PROJECT TOPIC- ANTIMICROBIAL ACTIVITY OF METHANOL EXTRACT AND FRACTIONS OF MORINGA OLEIFERA LAM. ROOT BARK ON CLINICAL ISOLATES OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS

 

ABSTRACT

Development of antimicrobial resistance by bacteria is now a world wide health issue, as infection is one of the leading causes of death in the world today. This fact is also as a result of the emergence of multiple antibiotic resistant bacteria known as methicillin resistant Staphylococcus aureus (MRSA) with potential of cross resistance to other antibiotics of choice like vancomycin. MRSA is often referred to as a potential killer and one of the tree top superbugs in hospitals multidrug resistant organisms (MDRO). The aim of this study was to evaluate the phytochemical components and antimicrobial activity of methanol extract and fractions of Moringa oleifera root bark as possible remedy for MRSA infections.
Staphylococcus aureus isolates from 3 different hospitals in South-east geopolitical region of Nigeria were confirmed by coagulase/staphylase test using Oxoid® reagents kits (DR0595A). The characterised S. aureus isolates were further identified as Methicillin resistant staphylococcus aureus by disc diffusion method as recommended by the Clinical Laboratory Standards Institute (CLSI), using standard antibiotic discs containing oxacillin (5 μg/ml), vancomycin (30 μg/ml), cephalexin (30 μg/ml), levofloxacin (5 μg/ml), ciprofloxacin (5 μg/ml), tetracycline (30 μg/ml), cotrimoxazole (25 μg/ml), gentamicin (30 μg/ml), clindamycin (2 μg/ml) and rifampicin (5 μg/ml). Methicillin resistant staphylococcus aureus confirmation was done using Oxoid® DR0900 penicillin binding protein (pbp2ˈ) latex agglutination test kits. Pulverised Moringa oleifera root bark was defatted with n-hexane to
yield hexane fraction (HEF). The dried marc was extracted with methanol using Soxhlet extractor to obtain crude methanol extract (ME). Methanol extract was adsorbed on Silical gel (60-200 mesh) and eluted in succession to obtain dichloromethane fraction (DMF), ethyl acetate fraction (EAF) and methanol fraction (MEF). Qualitative phytochemical analyses of the extracts were carried out using standard procedures. The antimicrobial activities of ME, HEF, DMF, EAF and MEF were evaluated on the MRSA, the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were recorded and compared with the standard disc antimicrobial test results. The extract fractions were analysed using gas chromatographic-mass spectrometry (GC-MS) for their bioactive compounds. The preliminary acute toxicity and sub-acute toxicity of ME and HEF were evaluated. Statistical analysis was done with ANOVA followed by Duncan post Hoc test using SPSS v 17 software. Characterised clinical isolates yielded 58 S. aureus strains. Antibiotic susceptibility tests indicated varied percentages of MRSA that were resistant to various antibiotics thus: oxacillin
(62.1 ± 3.2%), vancomycin (60.4 ± 3.8%), cephalexin (55.2 ± 1.2%), levofloxacin (56.9 ± 2.2%), ciprofloxacin (56.9 ± 0.9%), tetracycline (65.5 ± 2.3%), cotrimoxazole (68.9 ± 0.8%), gentamicin (67.2 ± 1.3%), clindamycin (62.1 ± 3.3%) and rifampicin (62.1 ± 4.1%). Latex agglutination test confirmed 39 strains of the clinical isolates to be MRSA. The S. aureus isolates resistant to all the antibiotics including vancomycin at 30 μg/ml were sensitive to the
extract and all its fractions: ME: MIC (3.0 ± 0.1 to 5.0 ± 0.5 mg/ml) and MBC (3.0 ± 0.1 to 6.0 ± 0.5 mg/ml); EAF: MIC (5.0 ± 1.1 to 8.0 ± 0.5 mg/ml) and MBC (5.0 ± 0.5 to 8.0 ± 0.5 mg/ml); DMF MIC (8.0 ± 1.1 to 10 ± 0.5 mg/ml) and MBC (8.0 ± 0.5 to 10 ± 0.5 mg/ml); HEF: MIC (7.0 ± 0.5 to 8 ± 1.1 mg/ml) and MBC (7.0 ± 0.5 to 9 ± 0.5 mg/ml), MEF: MIC (9.0 ± 1.1 to 10.0 ± 0.5 mg/ml) and MBC (9.0 ± 0.5 to 10.0 ± 0.5 mg/ml). Phytochemical analysis of the extracts showed the presence of alkaloids, glycosides, steroids, terpenoids, flavonoids, saponins, tannins, resins, reducing sugars, proteins, fats and oil and carbohydrates. GC-MS analysis revealed over 100 distinct compounds, some of which are stigmasterol (C29H48O), eugenol (C10H12O2), oxime (C3H7NO ) and ergosta-4, 22-dien-3-one (C28H44O). The oral acute toxicity test showed the LD50 of ME as 3663.96 mg/kg and HEF as 1934.15mg/kg, with no significant change (P > 0.05) in the hematological, serum biochemical parameters and weight of the rats.

CHAPTER ONE
1.0 Introduction

In the last few decades there has been an exponential growth in the field of herbal medicine. It is getting popularized in developing and developed countries owing to its natural origin and lesser side effects [1]. Herbal drugs constitute a major share of all the officially recognized systems of health in India viz. Ayurveda, Yoga, Unani, Siddha, Homeopathy and Naturopathy, except Allopathy. More than 70% of India’s 1.1 billion population still use these
non-allopathic systems of medicine [2]. In many developing countries, a large proportion of the population relies on traditional practitioners and their armamentarium of medicinal plants in order to meet health care needs. Although modern medicines may exist side-by-side with such traditional practice, herbal medicines have often maintained their popularity for historical and cultural reasons.

Such products have become more widely available commercially, especially in developed countries. Use of herbal medicines in developed countries has expanded sharply in the latter half of the twentieth century. In India, herbal drugs are an integral part of the Indian system of medicine (Ayurveda), which is an ancient and mainstream system [3].
The evaluation of various plant products according to their traditional uses and medicinal value based on their therapeutic efficacy leads to the discovery of newer and recent drugs for treatingvarious ailments. This fact forms the basis for the development of new drugs from various plant sources. One of such plants of medicinal value is Moringa olifera, belonging to the family Moringaceae, commonly known as ‘sahajan’ in Hindi, Horse radish in English. It is a small, fast, growing, evergreen, or deciduous tree that usually grows up to 10 or 12 m in height. It is distributed among Sub Himalayan Tracts, Assam, Bengal and Peninsular India [4]. Various properties are attributed to it like antispasmodic, diuretic, expectorant and abortifacient [5].

PROJECT TOPIC- ANTIMICROBIAL ACTIVITY OF METHANOL EXTRACT AND FRACTIONS OF MORINGA OLEIFERA LAM. ROOT BARK ON CLINICAL ISOLATES OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS

 

2 1.1 History of Moringa oleifera

It is a small or medium-sized tree, attractive enough to be a focal point in the tropics and sub-tropics owing to its creamy – white, sweetly scented flowers and light –green, tripinnately compound foliage [1-3]. It is a native to India, occurring wild in the sub- Himalayan regions of Northern India and cultivated throughout the country. It is commonly known as Sajina, sajna (Bengali); horseradish tree, drumstick tree (English); Sahinjan, mungna (Hindi); murinna, muringa, tishnagandha (Malyalam); sevaga, segata (Marathi); Sohanjana (Punjabi); Sobhanjana, sigru, murungi, dvishiguru (Sanskrit) and Sehjan(Urdu) in varied Indian languages and regions [4,5].

It also thrives well in Pakistan, Bangladesh, Sri Lanka, tropical Africa, Arabia, Philippines, Cambodia and Central, North and South America [6-10]. Described as “one of the most amazing trees God has created”, almost every part of drumstick viz. bark, root, fruit, flowers, leaves, seed and gum is a rich repository of proteins, vitamins and minerals including potassium, calcium, phosphorus, iron, folic acid as well as β carotene. Leaves can be eaten fresh, cooked or stored as dry powder for many months without refrigeration, without loss of nutritional value. Almost all the parts of this plant have been used for various ailments in the indigenous medicine of South Asia [11, 12].

The named varieties of moringa include Jaffna or Yazhpanam, grown in various parts of South India, (producing 60-90 cm long pods), Chavakacheri murungai, (producing pods 90-120 cm long), Chemmurungai (with red tipped fruits), Kadumurungai, Palmurungai, Puna murungai (with thick pulp and bitter taste), Kodikkal Murungai etc. [13,14]. The Horticultural College & Research Institute of Tamil Nadu Agricultural University has released two improved annual moringa varieties (PKM1, PKM2) within a span of 10 years, for commercial cultivation [15, 16].

The folklore claims and ancient literature report moringa to be an abortifacient antidote, antirheumatic, bactericide, diuretic, ecbolic, emetic, expectorant, purgative, rubefacient, stimulant, tonic, vermifuge and vesicant [17-20]. (Pharma Products Pvt Ltd, Thayavur, India) and Livospin (Herbals APS Pvt. Ltd., Patna, India), which are available for a variety of ailments [21]. Ayurvedic preparations include Ratnagiri Rasa, Sarasvata Ghrta, Sudarsana churna, Sarsapadi Pralepa, Visatimduka Taila etc
[4, 5]. Leaves of moringa are applied as poultice to sores and in treatment of anemia and menstrual irregularities. Young leaf paste with curd, is used internally for stomachache while externally for sprains. Leaf juice or bark paste is used as a drink for constipation and piles [22, 23]. The root juice is applied externally as rubefacient or counter irritant, in hiccups, lumbago, enlarged spleen or liver. Bark, leaves and roots are acrid and pungent, taken to
promote digestion. A reddish gum exuded from the bark possess anti diarrhoeal, emmenagogue, antiscorbic and abortifacient properties. According to Materia Medica, a compound spirit made from equal parts of roots of Moringa and orange peel acts as carminative and stimulant in nervous debility, paralytic afflictions, epilepsy and hysteria [24- 26]. Not only this, moringa is glorified as a ‘traditional mother care plant’, for the leaves are
highly nutritious for pregnant women [27].
Until now, only a very few attempts have been made to compile the myriad of potential uses of this “miracle tree”. In view of a number of recent findings of ethnopharmacological importance, an updated appraisal was much needed. So, the present research is an attempt to explore the claims so far and prepare the ground for development of effective novel herbal formulations of M. oleifera. in the treatment of infections caused by much dreaded Methicilin resistant Staphylococcus aureus [25-27].

1.2 Morphology

Moringa oleifera is a small, fast-growing evergreen or deciduous tree that usually grows as high as 9 m, with a soft and white wood and corky and gummy bark. Roots have the taste of horseradish. Leaves are longitudinally cracked leaves, 30-75 cm long main axis and Fig. 1 Compound leaf; paler lower surfaces of leaflets of Moringa oleifera Fig. 2 Flower panicle Moringa oleifera its branch jointed, glandular at joints, leaflets are glabrous and entire. The leaflets are finely hairy, green and almost hairless on the upper surface, paler and hairless beneath, with redtinged mid-veins, with entire (not toothed) margins, and are rounded or blunt-pointed at the apex and short-pointed at the base. The twigs are finely hairy and green. Flowers are white,
scented in large axillary down panicles, pods are pendulous, ribbed, seeds are 3-angled [4, 6].

1.3 Taxonomic classification

PROJECT TOPIC- ANTIMICROBIAL ACTIVITY OF METHANOL EXTRACT AND FRACTIONS OF MORINGA OLEIFERA LAM. ROOT BARK ON CLINICAL ISOLATES OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS

 

TO GET THE FULL PROJECT TOPIC AND MATERIAL DELIVERED TO YOUR INBOX OR DOWNLOAD INSTANTLY, PAY N5000 Via: BANK
BANKACCOUNT NAME
ACCOUNT NUMBER
DIAMOND BANK
FREEMANBIZ COMMUNICATION
007 031 2905
FIDELITY BANK
FREEMANBIZ COMMUNICATION
560 028 4107
GTBFREEMANBIZ COMMUNICATION013 772 5121
ZENITH BANK
FREEMANBIZ COMMUNICATION
101 326 3297
OR Pay Online with ATMAfter Payment, you can use the chat app at the right-hand side of your browser to download the material immediately or Text Name, Title of the project paid for, your email address to 08060755653.Do you prefer paying with Bitcoin, Ethereum, Bitcoin Cash or LiteCoin?

Hits: 19

Was the material helpful? Comment below. Need the material? Call 08060755653.

This site uses Akismet to reduce spam. Learn how your comment data is processed.