Synergistic Antibacterial Activity of Amoxicillin and Bakhaw (Rhizophora mucronata) bark crude methanolic and aqueous extracts against

Staphylococcus aureus and Escherichia coli

Buenafe, Genessa Dalisay, Janine Oto, Demi Monroy, Celestino Jegonia, Jasmine Marie Tagongtong, Vir Nigel Virgo, Angelie MLS 3B

Background and Rationale of the Study

 Infectious

diseases caused by bacteria affect millions of people worldwide. According to the World Health Organization (WHO), one-third of all deaths in the world are caused by infectious diseases Thus, the formulation and production of potent antibiotics against the infectious agents is a major undertaking for pharmaceutical institutions nowadays

 Commonly

used antibiotics have become less and less effective against certain illnesses. This is due to the increase use of broad spectrum antibiotics which allowed the bacteria to develop a mechanism towards its resistance. To address this problem, antibiotics are combined with other enhancing components acting in synergy towards the termination of the bacterial cell.

 One

approach to treat infectious diseases is the use of plant extracts individually or as an alternative approach is the use of a combination of antibiotics with plant extracts. One of the plants in the Philippines that show great potential in its phytochemical activity as an antibacterial agent are mangroves, specifically Rhizophora spp.

 As

such is the Stilt Mangrove or locally known as Bakhaw (Rizophora mucronata lamk.) which has been proven by previous studies that possess a potential antibacterial activity. Hence, this study looked on the possibility of a synergistic antibacterial activity of Amoxicillin and Bakhaw (Rhizophora mucronata) bark crude methanolic and aqueous extracts against Staphylococcus aureus and Escherichia coli.

Objectives

 Generally,

this research aims to identify the synergistic antibacterial activity of Amoxicillin and Bakhaw (Rhizophora mucronata) bark crude methanolic and aqueous extracts against Staphylococcus aureus and Escherichia coli.

Specifically, this research aims to:

1. To determine the zone of inhibition against Staphylococcus aureus and Escherichia coli of the different treatments:

Treatment A Crude Methanolic Plant Extract Treatment B Crude Aqueous Plant Extract Treatment C Amoxicillin Treatment D Crude Methanolic Plant Extract (10 g) + Amoxicillin (25 g) Treatment E Crude Methanolic Plant Extract (100 g) + Amoxicillin (25 g) Treatment F Crude Methanolic Plant Extract (1000 g) + Amoxicillin (25 g) Treatment G Crude Aqueous Plant Extract (10 g) + Amoxicillin (25 g) Treatment H Crude Aqueous Plant Extract (100 g) + Amoxicillin (25 g) Treatment I Crude Aqueous Plant Extract (1000 g) + Amoxicillin (25 g) Positive Control Co- Amoxiclav Negative Control Amoxicillin + distilled water

2. To determine if there is a significant difference in the mean zone of inhibition among different treatment concentrations as well as the positive and the negative control. 3. To determine if there is a significant difference in the mean zone of inhibition among treatment concentrations using two methods of extraction (Methanolic and Aqueous extraction). 4.To determine which among the treatments and mode of extraction shows the highest synergistic activity against Staphylococcus aureus and Escherichia coli.

Hypothesis

1. There is no significant difference between the different treatment concentrations as well as the positive and the negative controls based on the zone of inhibition.

Significance of the Study

 This

study aims to aims to identify the synergistic antibacterial activity of Amoxicillin and Bakhaw (Rhizophora mucronata) bark crude methanolic and aqueous extracts against Staphylococcus aureus and Escherichia coli. In this regard, this study is important to the people with great concerns about the multidrug resistance of E. coli and S. aureus:
benefit

hospitals and medical institutions pharmaceutical communities researchers as well as to students

Conceptual Framework

Independent Variables Different concentrations of the crude methanolic and aqueous plant extracts combined with amoxicillin.

Dependent Variables Diameter of the Zone of Inhibition

Scope and Delimitation

 This

study is limited only to identifying the synergistic antibacterial activity of Amoxicillin and Bakhaw (Rhizophora mucronata) bark crude methanolic and aqueous extracts against Staphylococcus aureus and Escherichia coli. The plant will be taken from Oton, Iloilo and Bakhaw, Jaro, Iloilo City, only. These mangroves will be identified by Dr. Junemie Ramos, mangrove expert, SEAFDEC AQD.

 The

study will be conducted at the University of San Agustin Research laboratory. Likewise, the positive control (AmoxiClav) as well as the negative control (Amoxicillin- distilled water solution) will be bought commercially. The Staphylococcus aureus and Escherichia coli for culture will also be purchased commercially. The statistical tool used in analyzing the data will be the Mean, Two Way Analysis of Variance and will be followed by the Post HoC analysis with a confidence level of 0.05.

Methodology

Plant collection

Plant identification

Processing of plant samples

Methanolic Extraction

Aqueous Extraction

Preparation of Treatment

Antibacterial Susceptibility Testing

Plan for Data Analysis

Proper Waste Disposal

Methodology