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INVESTIGATION ON THE INACTIVATION OF EXTENDED SPECTRUM BETA LACTAMASES USING VERNONIA AMYGDALINA (BITTER LEAF)

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Table of contents

 

CHAPTER ONE                                                                                                                     1 

1.1       Introduction                                                                                                                1 

1.2       Aim and Objective of the Study                                                                                 5 

CHAPTER TWO: Literature Review                                                                                    6 

2.1       Vernonia amygdalina Del. (V. amygdalina)                                                              6 

2.2       Extended-spectrum 𝛽-lactamases                                                                              12 

2.3       Beta Lactams                                                                                                              14 

2.4       Classification of Beta-Lactamases                                                                             15 

2.4.1    Group I (Ambler Class C) beta-lactamases (also known as AmpC enzymes)          15 

2.4.2    Group 2 (Ambler Class A) enzymes                                                                            16 

2.4.3    Group 3 (Ambler Class B) enzymes                                                                            16 

2.5       Types of ESBLs                                                                                                          17 

2.6       ESBL Producing Organisms                                                                                      19 

2.6.1    Enterobacteriaceae                                                                                                     19 

2.6.2    E.coli and Klebsiella species                                                                                      23 

2.6.3    Pseudomonas aeruginosa                                                                                           24 

2.6.4    Acinetobacter Species                                                                                                25 

2.7       Prevention and Control Management of ESBL                                                          26 

2.8       Treatment and Management of ESBL Infection                                                       27 

CHAPTER THREE: Materials and Methods                                                                         29 

3.1       Study Area                                                                                                                  29 

3.2       Test Samples and Method of Collection                                                                    29 

3.3       Collection and identification of the plant materials                                                   29 

3.4       Extraction of active components from plant materials                                               30 

3.5       Examination of Isolates for Multi-antibiotic resistance and ESβL- 

production                                                                                                                   30 

3.6       Effects of plant extracts on extended spectrum β-lactamase producing 

bacteria                                                                                                                       31 

3.7       Dilution of plant materials                                                                                          32 

3.8       Test isolates                                                                                                                32 

3.9       Antibacterial susceptibility testing of the plant extracts                                            32 

CHAPTER FOUR: Results                                                                                                    34 

CHAPTER FIVE                                                                                                                    42 

5.1       Discussion                                                                                                                   42 

5.2       Conclusion and Recommendation                                                                             44 

References                                                                                                                              46 

TABLE OF CONTENTS 

Table              Title                                                                                                    Pages 

1          Morphology and characterization of test organisms                                                  36 

2          Biochemical reaction of the test organisms                                                                37 

3          Inhibitory zones of the test organisms on extract at different concentration              38 

4          Minimum Inhibitory Concentration                                                                           39 

5          Minimum Bactericidal Concentration                                                                                    40 

6          Correlation between the Minimum Inhibitory Concentration and 

Minimum Bactericidal Concentration                                                                                    41 

LIST OF FIGURE 

Figure                    Title                                                                           Page 

1        Photos of V. amygdalina. 1a. V. amygdalina shrub. 1b. 1c. Flower 

of V. amygdalina. 1d. Leaves of V. amygdalina. amygdalina. 1d. 

 Leaves of V. amygdalina.                                                                11 

Abstract

 

The antibacterial activities of ethanolic extract of Vernonia amygdalina was performed against ESBL producing organisms obtained from Nigerian Institute of Medical Research, Yaba Lagos state which included Salmonella spp., Pseudomonas spp.  Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Serratia marcescens, Salmonella typhi, Proteus merabilis, Proteus vulgarius and Klebsiella oxytoca. The various inhibitory zone of the organisms at different concentration of the bitter leaf extract (6.2mg, 12.5mg, 25mg, 50mg, 100mg). Escherichia coli Serratia marcescens and Klebsiella pneumoniae had the highest zone of inhibition 21mm at 100mg, followed by Pseudomonas aeruginosa 19mm at 100mg, Salmonella spp. and Pseudomonas spp. 18mm at 100mg. All the isolates showed zone of inhibition at 50mg and 25mg but Pseudomonas spp., Proteus mirabilis and Proteus vugaris had no clear zone at 12.5mg. Only four organisms, Salmonella spp., Escherichia coli, Serratia marcescens and Salmonella typhi had zone of inhibition at 6.2mg. The minimum inhibitory concentration and minimum bactericidal concentration of the organisms at different concentrations of extract (100mg/ml – 0.20mg/ml). The MIC and MBC increase as the concentration increases. However, Escherichia showed highest susceptibility to the extract with 100mg/ml>100mg/ml while the least susceptible was Serratia marcescens with 3.13mg/ml>100mg/ml. Since it has been proved that the extract of V. amygdalina inhibit ESBL producing organisms at difference concentration, it could be used as broad spectrum antibiotics in the treatment of infections since it has antimicrobial activity. 

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