RESEARCH OUTPUT 10, 11: ACTUAL DATA ANALYSIS and DATA PRESENTATION

Adviser: Dr. I.A. Ilano Submitted by: GROUP 9-A Leader: Holgado, Anna Victoria

Members: Alcantara, Jan Christopher Balandan, Patricia Buenafe, Jonas Joaquin Constantino, Erwin Delos Santos, Kathrine Aira Flores, Marie Felle Hernandez, Kristeen Khae Lopez, Edison Date: February 15, 2012

Topic: "A comparative study on the antibacterial activity of the peel extracts obtained from Musa acuminata, Musa balbisiana, and Musa paradisiaca against Staphylococcus aureus

I.
A.

INTRODUCTION
Research question Which species of banana peel extract has greater antibacterial activity against Staphylococcus aureus? Research hypotheses i. Working hypothesis The peel extracts obtained from the different species of banana have comparable antibacterial activity against Staphylococcus aureus. ii. Null hypothesis The peel extracts obtained from the different species of banana do not have antibacterial activity against Staphylococcus aureus.

B.

II.
A.

RESEARCH OBJECTIVES and RESEARCH DESIGN

General objective To determine which species of banana peel extract has greater antibacterial activity against Staphylococcus aureus. B. Specific objectives 1. To measure the zones of inhibition of peel extracts obtained from Musa acuminata, Musa balbisiana, and Musa paradisiaca using the disk diffusion method. 2. To establish the respective minimal inhibitory concentrations of peel extracts obtained from Musa acuminata, Musa balbisiana, and Musa paradisiaca using broth dilution test. 3. To establish the respective minimal bacteriocidal concentrations of peel extracts obtained from Musa acuminata, Musa balbisiana, and Musa paradisiaca using broth dilution test. 4. To compare the obtained zones of inhibition, minimal inhibitory concentrations, and minimal bacteriocidal concentrations of the peel extracts against Staphylococcus aureus.

C.

Research design The study will utilize an analytic experimental design, wherein the independent

variable under observation will be assigned particularly to a treatment group and will be compared to a positive and negative control group. Figure 1 illustrates the design of the study.

Figure 1: Schematic diagram of research design

Operational definition of variables 1. Independent variable The antibacterial activity of the different species of banana peel extracts on the growth of Staphylococcus aureus serves as the independent variable for the study. Antibacterial activity refers to the capacity of an agent to kill or suppress the growth of microorganisms, specifically bacteria.(1) This property may be further classified into two mechanisms, bacteriostatic and bacteriocidal. Bacteriostatic activity results into the inhibition of microbial growth within a certain period of time. Microbial growth may be

observed once environmental elements become suitable, or the microorganism has gained resistance to counteract the stimulus presented by the agent.(1) On the other hand, bacteriocidal activity results into the complete eradication of the species. In the study, significant bacteriostatic activity of the different species of bananas will be observed through disk diffusion method and broth dilution test. Musa acuminata (locally known as lakatan), Musa balbisiana (saba), and Musa paradisiaca (latundan) are considered as the most common group of species grown and commonly sold in the province of Cavite. Because of their wide availability and easy accessibility, these species will be chosen as samples for the study. 2. Dependent variable The expected result from the study will be the inhibition of the growth of Staphylococcus aureus on nutrient agar medium. Inhibition denotes a temporary cessation in microbial growth processes. This implies that there are still possible chances for growth, given that the environment becomes favorable once again for microorganism propagation.(18) Therefore, strict compliance with the incubation of disk diffusion plates within the allotted time of 24 hours for the Staphylococci species should be observed so as to achieve reliable results and prevent possible growth of the organism.(2) Inhibition of growth will be determined qualitatively using the disk diffusion method, as represented by zones of inhibition. Broth dilution test will also be employed to quantify the degree of the antibacterial activity by determining the peel extracts minimal inhibitory concentration. The end point tube in the series of test tubes illustrates the absence of microbial growth achieved with the least concentration of the peel extract.
(1)(2)

Aliquots

from the tube with the least amount of drug that showed no growth and the two tubes that immediately precede it will be collected and inoculated in a nutrient agar medium using the streak plating technique in determining the peel extracts respective minimal bacteriocidal concetrations. 3. Confounding variables Both the independent and dependent variables may face possible contamination brought about by 1) their exposure to environmental factors, such as temperature and foreign body contamination; 2) as well as their subjection to certain preparation and storage techniques executed in the duration of the study. Contamination of the variables of interest may significantly influence the accuracy and analysis of the results. (1)(2)(3)

III.
A. i.

RESULTS AND ANALYSIS

Format for data analysis Descriptive statistics The mean diameter obtained among the three replicates will be calculated in measuring the central tendency of the zone of inhibition produced by the peel extract. Measures of central tendency are inappropriate for the values obtained in conducting MIC and MBC since these tests are performed only once. Furthermore, the standard deviation will be computed from the value gathered from the zone of inhibition. The final data will then be reported as mean + standard deviation. ii. Inferential statistics - One-way analysis of variance (ANOVA) will be used as the test statistic since the study is dealing with more than two independent study groups. This is based on the premise that each peel extract groups effect will be unique from the other as well as from the established control groups. One-way ANOVA is helpful in determining the probability of observing a difference existing among the means of several study groups at the same time, minimizes the possibility of committing a type I error which may result from conducting multiple t-tests.(4) The confidence interval for this study will be set at 95%, with the value for at 0.05. These values have been derived from previous studies on the antibacterial activity of plant extracts.(5) Table 1 shows the plan for data analysis that will be utilized in the study. The

objective is to compare the results of more than two independent groups. The mean diameter of the zones of inhibition obtained from each species of banana peel that will be compared is categorized under the ratio scale of measurement; therefore, the tool for data analysis that will be used will be a type of parametric test, namely, one-way ANOVA.

Table 1. Plan for data analysis

Plan for Data Analysis Variables Independent variable The antibacterial activity of the different species of banana peel extracts on the growth of Staphylococcus aureus Scale of measurement Ratio Descriptive? Inferential? 

Dependent variable - the inhibition of the growth of Staphylococcus aureus as measured by the zone of inhibition produced Variables are independently related to each other

Ratio

ANOVA

B.

Results i. Descriptive statistics The antibacterial activity of the peel extracts obtained from Musa acuminata, Musa

balbisiana, and Musa paradisiaca were initially determined using the disk diffusion method. Each species were prepared in three concentrations (30 g, 300 g, and 3,000 g) and were observed for their respective antibacterial activity as evidenced by the zones of inhibition produced. However, paper disk diffusion results revealed absence of inhibition in all species. Difficulty in replicating the methods from previous studies, wherein zones of inhibition were seen, brought about the need to conduct trials in determining the antibacterial activity of the peel extract. In each trial, the extract concentration used was set at 3,000 g. Several variables such as blending time, characteristics of the banana peel (e.g. color and storage duration), and the extract storage temperature were considered. Values for mean + standard deviation are then considered to be set at zero, since the values gathered per trial are null.(6) Table 2 shows the measurements of the zones of inhibition among the different banana species.

Table 2. Zone of inhibition (mm) of peel extracts of different banana species using paper disk diffusion in different trials TRIAL 1 Musa sapientum 0 TRIAL 2 0 TRIAL 3 0 TRIAL 4 0 TRIAL 5 0 TRIAL 6 0

Musa paradisiaca

Musa acuminata

ii.

Inferential statistics

Since the values obtained were set at zero, replacing these on the F-ratio (i.e. statistic used in utilizing ANOVA) would reveal that the p-value would progress towards a positive infinity, implying that the value of statistical significance is within the non-rejection area of the normal distribution curve.(7) A conclusion can then be made that there are no sufficient evidences to reject that the peel extracts obtained from the different species of banana do not have comparable antibacterial activity against Staphylococcus aureus, in favor of the alternative hypothesis.

REFERENCES:
(1) Mahon, C.R., Lehman, D.C., and Manuselis, G. (2007). Textbook of Diagnostic Microbiology. Missouri: Saunders. (2) Forbes, B.A., Sahm, D.F., and Weissfeld, A.S. (2007) Bailey and Scotts Diagnostic Microbiology. Missouri: Elsevier. (3) Department of Microbiology and Parasitology, DLS-HIS College of Medicine (2011). Laboratory Manual on Microbiology and Parasitology. (4) Analysis of Variance. Retrieved from http://www.wikipedia.com (5) Mokbel, M.S. and Hashinaga, F. (2005). Antibacterial and antioxidant activities of banana (Musa AAA., cv. Cavendish) fruits peel. American Journal of Biochemistry and Biotechnology, 3, 125-131. (6) MacMillan A., et al. (2007) Basic statistics: mean, median, average, standard deviation, z-scores, and pvalue. Retrieved from https://controls.engin.umich.edu (7) What is the meaning of an F value less than 1 in one-way ANOVA? Retrieved from http://stats.statexchange.com