ANTIMICROBIAL ACTIVITIES OF BRYOPHYLLUM PINNATUM LEAF EXTRACTS ON SPUTUM ISOLATES

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.

Buyers has the right to create dispute within seven (7) days of purchase for 100% refund request when you experience issue with the file received. 

Dispute can only be created when you receive a corrupt file, a wrong file or irregularities in the table of contents and content of the file you received. 

ProjectShelve.com shall either provide the appropriate file within 48hrs or send refund excluding your bank transaction charges. Term and Conditions are applied.

Buyers are expected to confirm that the material you are paying for is available on our website ProjectShelve.com and you have selected the right material, you have also gone through the preliminary pages and it interests you before payment. DO NOT MAKE BANK PAYMENT IF YOUR TOPIC IS NOT ON THE WEBSITE.

In case of payment for a material not available on ProjectShelve.com, the management of ProjectShelve.com has the right to keep your money until you send a topic that is available on our website within 48 hours.

You cannot change topic after receiving material of the topic you ordered and paid for.