ABSTRACT
The antimicrobial activities of the leaf extracts of Bryophyllum pinnatum on sputum isolates were studied. Organisms obtained from the culture of the infected sputum includes Streptococcus pneumonia, Staphylococcus aureus, Klebsiella pneumonia, Pseudomonas aeruginosa and a fungal yeast (Candida albicans). Bioactive compounds were extracted from the plant leaves using different solvents (Aqueous, methanol, ethanol and acetone). Preliminary phytochemical analysis reveals the presence of Alkaloid, Saponin, Tannin, Cyanogenic Glycoside, flavonoid, Steroid and Phenol. Antioxidant activities obtained were Scavenging at concentration of 54.3%, Chelating at 45.8% and Reducing at concentration of 30.83%. Using disc diffusion method each prepared extract was tested on the different isolated organisms and the extent of sensitivity of the tested organism on the different extracts was measured by their zones of Inhibition. Methanol extract inhibited growth of the test organisms at the highest range of 12.33 to 23.67mm, Ethanol at the range of 11.33 to 23.00mm, Acetone at the range of 10.00 to 20.00mm and Aqueous extract at the range of 12.33 to 17.33mm. A standard broad spectrum antibiotic (Ciprofloxacin) was use as a control. The action of the different extracts shows variation of significant differences (P<0.05). The lowest Minimum Inhibition Concentration (MIC) of the extracts was at concentration of 100mg/ml and the highest Minimum Inhibition Concentration was at 200mg/ml. There is basis for the traditional use of extracts of Bryophyllum pinnatum plant as a local health remedy for respiratory tract infections and the result of this study can be a precursor for the production of drugs for respiratory tract infections.
TABLE OF CONTENT
Title page
Certification i
Dedication ii
Acknowledgement iii
Table of content iv
List of Tables ix
List of Figures
x
Abstract xi
CHAPTER ONE
1.0 INTRODUCTION 1
1.1 Aims and objectives 3
CHAPTER TWO
2.0 LITERATURE REVIEW 4
2.1 Medicinal plant 4
2.2 Bryophyllum
pinnatum plant 5
2.3 Phytochemical Constituent of Bryophyllum pinnatum 8
2.4 Medicinal and pharmacological value of Bryophyllum pinnatum 10
2.5 Antioxidant properties of Bryophyllum
pinnatum 11
2.6 Antimicrobial Agents 13
2.6.1 Characteristics of Antimicrobial Agent 13
2.6.2 Factors Influencing the Effectiveness of
Antimicrobial Agent Activity 13
2.7 Sputum 14
2.7.1 Types of Sputum 14
2.7.2 Common Pathogens Associated with Sputum 15
2.8 Respiratory Tract Infections 15
2.8.1 Upper Respiratory Tract Infection 16
2.8.2 Lower respiratory tract infection 16
CHAPTER THREE
3.0 MATERIALS
AND METHODS 17
3.1 Materials 17
3.2 Collection of plant sample 17
3.2.1 Preparation of sample for extraction 17
3.3 Production of Extracts 18
3.4 Sputum sample Collection 18
3.5 Media preparation 18
3.6 Isolation, characterization and
identification of the isolates 19
3.6.1 Purification of isolates (Subculture) 19
3.6.2 Characterization of isolates 19
3.6.2.1 Characterization of fungi isolate 20
3.6.2.2 Characterization of bacteria isolates 20
3.7 Gram Stain Reaction 21
3.8 Biochemical tests 21
3.8.1 Sugar tube fermentation test 21
3.8.2 Catalase test: 22
3.8.3 Motility test 22
3.8.4 Urease test 22
3.8.5 Indole test 22
3.8.6 Oxidase test 23
3.8.7 Methylene red 23
3.8.8 Coagulase test 23
3.8.9 Citrate utilization test 24
3.9 Phytochemical analysis 24
3.9.1 Qualitative analysis 24
3.9.1.1 Test for Tannins 24
3.9.1.2 Test for Saponin 25
3.9.1.3 Test for Flavonoid 25
3.9.1.4
Test for Alkaloid 25
3.9.1.5
Test for Steroids 26
3.9.1.6
Test for Phenol 26
3.9.2 Quantitative content of the phytochemical 26
3.9.2.1
Determination of Alkaloid 26
3.9.2.2
Determination of Flavonoid 27
3.9.2.3
Determination of Tannin 28
3.9.2.4
Determination of Saponin 29
3.9.2.5
Determination of Phenol 30
3.9.2.6 Determination of
Cyanogenic Glycoside 31
3.10 Determination of Antioxidant activity 32
3.10.1 Determination of scavenging activity 32
3.10.2 Ferrous
ion-chelating ability 33
3.10.3 Ferrous reducing antioxidant power 33
3.11 Antimicrobial assay 33
3.11.1 Determination of Minimum Inhibition
Concentration 34
3.12 Statistical Analysis 34
CHAPTER FOUR
4.0
RESULTS 35
CHAPTER FIVE
5.0 DISCUSSION,
CONCLUSION AND RECOMMENDATION 43
5.1 Discussion 43
5.2 Conclusion 45
5.3 Recommendation 45
REFERENCE
APPENDICES
LIST OF TABLES
Table Title Pages
1 Taxonomial
Classification of Bryophyllum pinnatum 6
2 Identification
of bacteria isolated from sputum 37
3 Isolation
and identification of fungi isolated from sputum 38
4 Qualitative
phytochemical screening of Bryophyllum
pinnatum extracts 39
LIST OF FIGURES
Figure Title Pages
1 Picture of Bryophyllum pinnatum plant 7
2 Antioxidant activities of the extracts
of Bryophyllum pinnatum leaves (%) 40
3 Antimicrobial activities of extracts
of Bryophyllum pinnatum 41
4 Minimum inhibition concentration
(mg/ml) of the extracts 42
CHAPTER ONE
INTRODUCTION
Bryophyllum pinnatum belong
to Crassulaceae family with a pan-tropical distribution. It is a smooth robust
and unbranched herb that grow 30-200cm tall, it is also called “wonder of the
world” in English speaking Caribbean. According to Burkill 1995, Bryophyllum pinnatum has been of upmost
important in the treatment of respiratory disorder. This is as a result of the
fact that the plant leaf contain, potassium, insulin, vitamins as well as other
compounds such as saponin, flavonoid, alkaloids,
triterpenes, glycosides, flavonoids, steroids, Anthraquinoes, Xanthones,
Anti-inflammatory hypoglycaemic, anti-diabetic, anti-oxidant and anti-cancer
properties (Okwu and Josiah. 2006), also the extract of this plant possess
medicinal properties such as; immune-modulator, CNS depressant analgesic,
anti-inflammatory. Antiallergenic, anti-ulcerous, gastro-protective,
insecticidal and sedative (Akinpelu., 2000; Pal et al., 1991). In rural areas, the juice obtained from Bryophyllum pinnatum are used by the
people in the treatment or remedy for chronic cough. They could also be applied
to induce vomiting of blood, cut umbilical cord in new born, ear-ache,
diarrhea, ulcer abscesses, whitlow, insect bite, chronic bronchitis and other
infections associated with Respiratory tract infections (Burkill 1995). The
juices from fresh leaves is used to treat smallpox, headache, convulsion and
general debility. Some biological
activities of Bryophyllum pinnatum have
been reported to have anti-microbial potential in the treatment of bacterial,
fungal or viral isolate from infected respiratory tract sample, the plant possess antimicrobial
potential which are effective in combating of some common isolate from sputum
such as Staphylococcus aureus,
Escherichia coli, Pseudomonas aeruginosa, Klebsiella aerogenes, Streptococcus
spp etc (Ofokani et al., 2005)
Bacterial resistance to antibiotics is becoming a concern to
public health. The increasing resistance of many microorganisms to many
antibiotics has prompted the searches for new antimicrobial and antifungal
agents either by synthesis of new agents or through the
emergence of new pathogenic strains that are multi –resistance to antibiotics
(Aibinu et al., 2003; Aibiau et al., 2004). The non-availability and
high cost of new generation antibiotics with limited effective span have
resulted in need to look at other substances from other sources with proven
antimicrobial activity. Consequently, this has led to the research for more
effective agents among materials of plant origin with the aim of discovering
potential useful active ingredients that serves as a source and template for
the synthesis of a new antimicrobial drug. The Plants selected and empirically
use as drugs in alternative for antibiotics has initially been use as
traditional preparation then as pure active principle, with this, knowledge and
accumulation practice has passed from generation to generation (Taylor et al., 2001). Many of these plants
contain some bioactive principles which have not been fully discovered.
Traditionally, herbs and herbal products have been considered to be non-toxic
and have been used by the general public and traditional medicinal
practitioners worldwide to treat a range of ailment. The active ingredient of
plant extract are chemicals that are similar to those in purified medication,
and they have the same potential to cause serious adverse effects. The
potential for developing antimicrobial agents from higher plants appears
rewarding as it lead to developing of phytomedicine to act against microbes.
Plant base antimicrobial have enormous therapeutic potential as they can serve
the purpose with lesser side effects than those associated with synthetic
antimicrobial agents.
Infected
sputum contains respiratory pathogens which can easily be spread through air.
They cause series of respiratory tract infection with effects such as bronchial
pains, severe dry cough etc.
Phytochemical
substances mostly of this plant based sources have been of upmost important in
combating different forms of respiratory disorder (Mudi and Ibrahim 2008).
1.1
Aims
and objective
·
To identify common
respiratory tract pathogens in sputum.
·
To investigate the
sensitivity and resistivity of Bryophyllum
pinnatum leaf extracts on selected sputum isolate.
·
To compare the
antimicrobial potential of Bryophyllum
pinnatum extracts with a selected broad spectrum antibiotic.
·
To investigate the
phytochemical constitute of Bryophyllum
pinnatum plant.
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