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Apple, Guava and Pineapple Fruit Extracts as Antimicrobial Agents Against Pathogenic Bacteria

Samiha Kabir
Biotechnology Programme, Department of Mathematics and Natural Sciences, BRAC University, Mohakhali 66, Dhaka, Bangladesh

Sheikh Mehbish Jahan
Biotechnology Programme, Department of Mathematics and Natural Sciences, BRAC University, Mohakhali 66, Dhaka, Bangladesh

M. Mahboob Hossain
Biotechnology Programme, Department of Mathematics and Natural Sciences, BRAC University, Mohakhali 66, Dhaka, Bangladesh

Romana Siddique*
Biotechnology Programme, Department of Mathematics and Natural Sciences, BRAC University, Mohakhali 66, Dhaka, Bangladesh

Abstract/ Executive Summary:

This investigation was conducted to compare the antimicrobial properties of alcohol fruit extracts from Apple (Malus pumila), Guava (Psidium guajava), and Pineapple (Ananas comosus) against eight bacteria: Staphylococcus aureus, Enteroaggregative E.coli (EAEC), Enterotoxigenic E.coli (ETEC), Enterobacter cloacae, Shigella flexineri, Enterococcus faecalis, Klebsiella, and Pseudomonas aeruginosa. All the fruits were bought fresh from the market, cut into small cubes, and sun dried to a crisp over 4-5 days. After being blended to a fine powder, 75g of each powder was passed through a Soxhlet apparatus containing 250 ml of 99% ethanol to obtain a crude extract. This process was repeated with 250 ml of methanol to obtain a methanolic crude extract. The antimicrobial properties of the extracts were tested using agar well diffusion, Norfloxacin used as positive control and water as negative control. The inhibition zones from each extract were measured and an activity index was calculated from the mean zone sizes. All fruits showed some degree of antimicrobial properties with the highest activity index (2.6) being from Pineapple ethanolic and methanolic extracts against EAEC. Both Apple ethanolic and methanolic extracts showed activity only against Staph. aureus and EAEC. Guava extracts were effective against all the strains except for ETEC. Pineapple extracts were strongly active against all the bacteria. The methanolic extracts of Apple and Guava showed slightly larger zones compared to their ethanolic extracts. For Pineapple, inhibition zones from the ethanolic extracts were slightly larger. The results of this investigation show great promise for potential antimicrobial drugs.

Keywords: Soxhlet, Activity index, Crude extract, Well diffusion

Location: Biotechnology Programme, Department of Mathematics and Natural Sciences, BRAC University, Mohakhali 66, Dhaka, Bangladesh

Start Date:

End Date:

Program Area: Pest Management

Commodity: Apple, Guava, Pineapple

Objectives:

Bromelain is also a strong antifungal agent, inhibiting the growth of F. verticillioides by 90%, at only 0.3 micromoles per litre. Juice from Pineapple has also exhibited antibacterial action against E. coli, Shigella sonnei and Salmonella para.B. There, this study was undertaken to verify the objectives.

Methodology:

2.1. Strains of Bacteria The following strains of bacteria were used: Staphylococcus aureus, Enteroaggregative E. coli (EAEC), Enterotoxigenic E. coli (ETEC), Enterobacter cloacae, Shigella flexineri, Enterococcus faecalis, Klebsiella, and Pseudomonas aeruginosa. All strains of bacteria were maintained in laboratory fridge through regular subcultures.

2.2. Preparation of Extracts Fruits were bought fresh from local market and cut into small cubes. Seeds were carefully removed. The fruit cubes were kept under direct sunlight for about 4-5 days until they lost all moisture and became dried to a crisp. After that, the cubes were put in a blender and turned to a fine powder. Seventy five grams of finely ground powder was measured, placed in a thimble and run through a Soxhlet apparatus. The flask in the Soxhlet was filled with 250 ml absolute ethanol and 250 ml methanol as solvents, individually, to obtain ethanolic and methanolic crude extracts respectively. The cycles were continued for about 4-5 hours until the cotton inside turned white.

2.3. Agar Well Diffusion Fresh subculture plates, incubated for 24 hours, were placed under the laminar chamber and used to make standard bacterial suspensions in labeled test tubes. Next, an autoclaved cotton swab was used to perform lawn culture for the uniform growth of bacteria. After that, a cork borer was dipped into the agar to make three holes or wells in the media. Each well was labelled and accordingly filled with 30 microlitres of an ethanol fruit extract, methanol fruit extract and water for negative control. Norfloxacin, an antibiotic disc, is also placed in the dish in the form of positive control. The plates are incubated for 24 hours at 37°C, after which clear zones were formed around the control disc and the extracts which gave positive results. These inhibition zones were measured in millimetres using a ruler and recorded. All antimicrobial tests were repeated twice and the average of the inhibition zones is noted. An activity index was calculated from the results to measure the relative efficacy of the fruit extracts. The following formula was used:

Activity Index (AI) = Zone of inhibition of fruit extrac / Zone of inhibition of Norfloxacin

Source of Information: American Journal of Microbiological Research, 2017, Vol. 5, No. 5, 101-106

Article Link (Online Journal): DOI:10.12691/ajmr-5-5-2

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Though the use of fruits in medicine has been employed over thousands of years, it is only recently that scientific research has taken an interest in them. Antibiotic resistance is a growing global issue. Especially in our country, where more than half the population is not educated, this is an even bigger problem. Antibiotics are bought and sold without prescriptions. Often they are prescribed unnecessarily and the courses are not completed. Consequently, many resistant strains of bacteria have also been reported in Bangladesh. Using fruits to treat diseases could greatly decrease the pressure on antibiotics. Also, in the context of Bangladesh, health care is inaccessible to many. Eating fruits that are readily available and relatively cheaper could greatly help the people.

Knowledge Management: Journal