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Microbiological Analysis and Determination of Antimicrobial Traits of Green Banana (musa Spp.) and Papaya (carica Papaya)

Nusrat Jahan
Department of Microbiology, Stamford University Bangladesh, 51 Siddeswari Road, Dhaka 1217, Bangladesh

Rashed Noor*
School of Life Sciences (SLS), Independent University, Bangladesh (IUB), Plot 16, Block B, Aftabuddin Ahmed Road, Bashundhara, Dhaka 1229, Bangladesh

Saurab Kishore Munshi
Department of Microbiology, Stamford University Bangladesh, 51 Siddeswari Road, Dhaka 1217, Bangladesh

Abstract/ Executive Summary:

Green banana (Musa spp.) and papaya (Carica papaya), which are commonly consumed as vegetables, are fruits with lots of health benefits. Due to their various medicinal applications, both the fruits are likely to contain antimicrobial properties. In this regard, present study was undertaken to determine the microbiological quality as well as the anti-bacterial traits of fresh green banana and papaya. To serve the purpose, a total of 12 samples (6 green bananas and 6 green papaya) were collected from different local and super shops as well from cultivation land of some villages of Bangladesh. The samples contained the microbial contamination with viable bacteria and fungi up to 107 cfu/g and 103 cfu/g. Among the pathogenic bacteria, Staphylococcus spp. Pseudomonas spp. and Vibrio spp. were predominant as found in most of the samples in average of 102 cfu/g. Whereas Escherichia coli, Salmonella spp. and Shigella spp. were totally absent in all the samples. On the other hand, Klebsiella spp. was present in same samples. Additionally, the anti-bacterial poverties of the samples were chalked out. The in-vitro antimicrobial activities of the ethanolic, methanolic and hot water extracts of the samples were noticeably found against all the tested bacteria. Overall, present study revealed that both the green fruit samples were highly contaminated although they have antimicrobial activities. Thus, consumers’ awareness on the risk of consuming pathogen contaminated vegetables needs to be re-awakened and further investigations are required for potential antimicrobial properties.

Keywords: Green banana; Green papaya; Microbiological analysis; Antibacterial activity

Location: Microbiology Laboratory of Stamford University Bangladesh

Start Date: 01-03-2017

End Date: 04-04-2017

Program Area: Resource Development and Management

Commodity: Banana, Papaya

Objectives:

The study was carried out to investigate the microbiological quality and antimicrobial traits of fresh green bananas and papaya. Samples were collected from different areas in Bangladesh and tested for the presence of antibacterial activity through agar well diffusion method following ethanolic, methanolic and hot water extraction.

Methodology:

Study area and sampling. A total of 12 green bananas and papaya samples, 4 from local market in Dhaka city, 4 from village market and 4 from super shops were randomly collected from different types of shops. All the samples were prepared, processed and analyzed during the period of 1st March 2017 to 6th April 2017 in Microbiology Laboratory of Stamford University Bangladesh. Sample processing. The samples were processed following standard methods. 25 gm of green fruit samples weighted and added into 75 ml of normal saline in sterile blender and a homogenized mixture of sample as prepared to obtain 10-1 dilution of the sample shake well by vortex mixture. Then 1 ml from 10-1 dilution was transferred to 9 ml of normal saline to make 10-2 dilution and using the same procedure, sample was diluted to 10-5 dilution (Cappucino and Shermen, 1996, Sharmin et al. 2014, Das et al. 2017, ; Acharjee et al., 2014, Mrjan et al., 2014, Ahmed et al., 2013; Noor et al. 2013). By following serial dilution technique the blend samples were spread on different types of media for enumeration of bacterial load. Inoculation and enumeration of microorganisms. Spread plate technique was used to inoculate the samples on various culture media for enumeration of microorganisms. Melted medium is poured in a petri dish and allow solidifying. For the growth of total viable bacteria and fungi, 0.1 ml of samples from 10-3 and 10-5 dilutions was added to nutrient agar medium (NA) and Sabouraud’s dextrose agar (SDA) plates. From dilution 10-3 , 0.1 ml sample was added to MacConkey agar (MAC), Mannitol salt agar (MSA) and Pseudomonas agar (PA) for the isolation and enumeration of specific bacterial pathogens. All media were incubated at 37 oC for 24 hours and SDA were incubated at 25 oC for 24-48 hours for total viable fungi (23-29). For the enrichment of Salmonella, Shigella, and vibrio spp., 10 ml of sample was transferred into 90 ml of selenite cysteine broth (SCB) and alkaline peptone water (APW), respectively and incubated at 37 oC for 6 hours (30-33). After incubation , the samples were diluted up to 10-6 and then 0.1 ml of samples from the dilution 10-3 and 10-5 of each of the samples was spread onto Salmonella-Shigella (SS) agar and thiosulfate citrate bile salt sucrose (TCBS) o (TCBS) agar for the isolation of Salmonella spp. and Shigella spp., and Vibrio spp., consecutively. Plates were incubated at 37 oC for 24 hours for the detection of typical colonies (23-29). Preparation of the samples for extraction and conduction of antimicrobial assay. The dried powder form of samples were prepared through grinding and added 10 g of powder in 100 ml of ethanol and methanol in Durham’s bottle which were kept in shaking water bath at 130 rpm for 24 h. at 24 °C. Afterward the extract solution was filtered and the pellet of the samples were collected to observed their anti-bacterial properties against different pathogenic strain (34) by agar well diffusion method (24, 30, 35, 36). At first the lawns of bacterial suspensions (105 cfu or 0.5 OD measured by spectrophotometer) including Escherichia coli, Pseudomonas spp., Shigella spp., Vibrio spp., Klebsiella spp., Staphylococcus aureus, and Salmonella spp. were prepared and 100μl of the hot water extract, ethanol- and methanol extracts at a concentration of ~11.1mg/ml each were introduced into the wells. Absolute ethanol and methanol were used as negative controls while the antibiotic discs of gentamicin (10 μl) were used as positive control. Plates were incubated at 37 °C for 12-18 hours and examined for formation of the zone of inhibitions (mm).

Source of Information: Stamford Journal of Microbiology, 2018. Vol. 8, Issue 1, p. 41-45 ISSN: 2074-5346 (Print); 2408-8846 (Online)

Article Link (Online Journal): DOI: https://doi.org/10.3329/sjm.v8i1.42439

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Presence of pathogenic bacteria in extensive amount in fresh fruits or vegetables is not desirable in any condition which may cause food-borne illness in human. Government should take some necessary steps in agriculture sector to reduce the health risk factors by food-borne illness. Collectively, all the samples exhibited significant antibacterial activity as revealed from agar well diffusion method. However, presence of anti-bacterial traits of the samples ensured presence of antibacterial substance in fruits samples. Despite the significant progress made in microbiology and the control of microorganisms, drug resistant microorganisms have become a major threat to public health. The use of medicinal fruits or vegetables with antimicrobial activity requires more attention to combat the situation.

Knowledge Management: Journal