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 ﾿