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Antibacterial activities of plantain (Musa paradisiaca) peel and fruit

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Der Pharmacia Lettre, 2016, 8 (5):5-11

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ISSN 0975-5071
USA CODEN: DPLEB4

Antibacterial Activities of Plantain (Musa paradisiaca) Peel and Fruit

Asoso O. S., Akharaiyi F. C.* and Animba L. S.

Department of Biological Sciences, Afe Babalola University, Ado-Ekiti, Ekiti State, Nigeria
_____________________________________________________________________________________________

ABSTRACT

Plantain (Musa paradisiacae) is used as a medicinal plant employed in traditional system of healing diverse
diseases such as hepatitis, skin infections, problems concerning the digestive organs, respiratory organs,
reproduction, the circulation, anti-inflammatory, antiviral, analgesic, antioxidant, anti-carcinogenic, antitumor,
anti-nociceptive (reducing the sensitivity to painful stimuli), weakly antibiotic, immune modulation, anti-
ulcerogenic, anti-leukemic and antihypertensive effects, and for reducing fever. The antimicrobial activities of
methanol, ethanol and acetone extracts of M. paradisica peel and fruit were tested in-vitro against seven typed
Gram negative and positive pathogenic bacteria (Salmonella typhi 22648 ATCC, Salmonella typhi 23456 ATCC,
Escherichia coli 35218 ATCC, Shigella dysentrariae 24162 ATCC, Klebsiella pneumonia 34089 ATCC,
Staphylococcus aureus 25923 ATCC and Bacillus subtilis 21332 ATCC. The clinical isolates are Staphylococcus
aureus, Escherichia coli and Salnomella typhi. The antibacterial activity was assessed by agar well diffusion
method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values. M.
paradisica peel methanol and ethanol extract showed a higher zone of inhibition of test organisms than M.
paradisica fruit methanol and ethanol extract which could be due to phytochemical constituents. Phytochemical
analysis of the peel and fruit indicated the presence of alkaloid, flavonoids, saponins, tannins, phlobatannins,
glycosidea, and terpenoids. M. paradisica acetone extract from both fruit and peel showed no antibacterial
activities towards the organisms used.

Keywords: Antibacterial, clinical bacteria, Musa parasidiaca, inhibition

_____________________________________________________________________________________________

INTRODUCTION

Medicinal plants, as source of remedies, are widely used as alternative therapeutic tools for the prevention or
treatment of many diseases [1]. Plants are a viable, unlimited source of bioactive molecules, including antimicrobial
agents which protect them from microorganism, insects, and predators [2, 3, 4]. The use of medicinal herbs in
traditional system of medicine is a common practice in many cultures around the world especially in African
societies. This practice has gained widespread acceptance in developing as well as in developed nations. Researchers
are also beginning to appreciate the role of medicinal plants in health care delivery [5]. In recent time, interest with
herbal medicine for antimicrobial activities has been increased significantly. This is as a result of the effectiveness,
low cost and the availability of these herbal medicines, the economic crisis, high cost of industrialized medicines,
inefficient public access to medical and pharmaceutical care, in addition to the side effects caused by synthetic drugs
are some of the factors contributing to the central role of medicinal plants in health care [6, 7].

Plantain (Musa paradisiacae) is a major food crops in the humid and sub-humid parts of Africa where its starchy
fruits are generally cooked or fried before consumption and serves as major sources of energy for millions of people

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in these regions [8]. It belongs to the natural order, plantaginaceae which contains more than 200 species, twenty-
five or thirty of which have been reported. The common plantain has broad, irregular oval leaves, abruptly
contracted at the base into a long broad, channelled foot stalk. The fully grown blade is 1.3–2.4 meters long and
about two- third as broad, usually smooth, with several parallel veins. Plantain grows more than any other plant in
compacted soils, is abundant beside paths, roadside and other areas with frequent soil compaction. It is also common
in grassland and as a weed among crops.

Since unripe plantain flour is used by the traditional medical practitioners in Nigeria for dietary management of
diabetes mellitus and other disease conditions, this study is therefore aimed at investigate the antibacterial actives of
unripe plantain flour derived from fruits and peels.

MATERIALS AND METHODS

Collection of plantain samples

Fresh unripe plantains were obtained from Garki market, Abuja, Nigeria. The peel and fruit (unripe) were removed
by hand and cut into smaller pieces for easy drying. The dried peel and fruit were ground using a milling machine.
The powdery samples were packed into screwed bottles and labelled appropriately.

Collection of test organisms (bacterial strains)

Test bacterial strains used in the study are clinical and typed isolates were kindly provided by the microbiology
department in National Institute of Pharmaceutical Research and Development (NIPRID) Abuja, Nigeria. The test
isolates are Staphylococcus aureus, Escherichia coli, Salmonella typhi, Staphylococcus aureus (25923), Bacillus
subtilis (21332), Escherichia coli (35218), Samonella typhi (22648), Shigella dysentrariae (24162), Salmonella
typhi (23456), and Klebsiella pneumonia (34089).

Extraction of plantain peel and fruit

One hundred and twenty grammes of each powder were extracted with methanol, ethanol and acetone for 72 hours
with intermittent shaken in a shaking water bath. It was filtered through Whatmann No filter paper. The extracts
were evaporated under reduced pressure at 45 oC using a rotary evaporator.

Antibacterial Activities of extracts

One millilitre of each test isolate prepared to McFarland standard was ascetically pour plated with freshly prepared
Mueller-Hinton agar. The seeded plates were stand for 2 hours before wells bored into the agar using a sterile cork
borer. The residual extracts were dissolved in 5% Dimethyl Sulphur Oxide (DMSO4). Thus, 100µl of the different
extract were placed into the wells and the plates were incubated at 37 °C for 24 – 48 hours. Antibacterial activity of
extracts was evaluated by measuring the diameter of circular inhibition zones around the well. Tests were performed
in triplicate.

Antibiotic Susceptibility Test

The antibiotic susceptibility test was done using the agar-disc diffusion method. Molten Mueller Hinton agar cooled
to about 45 °C was poured into sterile petri dishes. And 1 ml of cell suspensions prepared in comparison to 0.5
McFarland standard was added and spread evenly onto the surface of the agar plates using sterile swap sticks. The
antibiotic disc was placed aseptically on the agar surface (the positive disc for the gram positive and the negative
disc for the gram negative) and plates were incubated at 37 °C for 24 hours. The zone of inhibition was then
measured and recorded. The tests were done in duplicate to ensure reliability [9].

Minimum Inhibitory Concentration (MIC) and minimum bactericidal Concentration (MBC)

Minimum inhibitory concentration (MIC) was determined for each extract showing antimicrobial activity against the
test isolates using broth micro dilution method. The MIC values were taken as the lowest concentration of the
extracts in the well of the test tube that showed no turbidity after incubation. Turbidity of the wells was interpreted
as visible growth of organisms. The minimum bactericidal concentration (MBC) was determined by sub culturing
from each well showing no growth. Least concentration of the extract showing no visible growth on sub culturing
was taken as MBC [10].

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RESULTS

All the test organisms were susceptible to the solvent extracts of M. paradisiacal peel and fruit. (Tables1a and 1b).
Among the solvent extraction, methanol extract was the most potent on the test organisms followed by the ethanol
and acetone extracts. However, the most inhibited test isolates include Salmonella typhi, Bacillus subtilis,
Escherichia coli, and Shigella dysentriae, which were inhibited with 35, 37, 40, and 30 mm by the methanol extract
of plantain peel and 27 and 22 mm by ethanol extract of peel while the acetone extract had little or no effect on the
organisms. These organisms were also inhibited by the fruit extracts of methanol with 27, 27, 20 and 27mm while
the ethanol and acetone extracts had little or no effect on test organisms respectively. The inhibition on the test
organisms by the solvent extracts of plantain peel and fruit are comparable to the inhibition by both positive and
negative control antibiotic (Figures 1 and 2). Figure 1 and 2 expresses susceptibility of test organisms to commercial
antibiotic (positive and negative control) among the commercial antibiotics inhibited. It was observed that all the
Gram positive organisms were resistant to ceftazidime, augmentin, cloxacillin, and ofloxacin while Staphylococcus
aureus (clinical isolate) was resistant to erythromycin and Bacillus subtilis (ATCC 21332) was resistant to
cefuroxime. All Gram negative organisms were resistant to cefuroxime, and augmentin, Escherichia coli (ATCC
35218) was resistant to ceftazidime, cefuroxime, gentamicin, cefixime, ofloxacin, augmentin, nitrofuration, and
ciprofloxacin. Shigella dysentrariae (ATCC 24162) was resistant to ceftazidime, cefuroxime, and augmentin. In
comparison of the plant extract with commercial antibiotics, the test isolates in most cases were highly susceptible to
the plant extract than the commercial antibiotics.

Table 1a: Antibacterial activities of the peel of Musa paradisica Zone of inhibition (mm)

Organisms Methanol Ethanol Acetone

Escherichia coli 40 ± 0.0 27 ± 0.0 12 ± 0.0
Escherichia coli ATCC 35218 22 ± 0.0 17 ± 0.0 -
Staphylococcus aureus 22 ± 0.0 17 ± 0.0 12 ± 0.0
Staphylococcus aureus ATCC 25923 22 ± 0.0 17 ± 0.0 -
Salmonella typhi 35 ± 0.0 27 ± 0.0 -
Salmonella typhi ATCC 22648 27 ± 0.0 17 ± 0.0 -
Salmonella typhi ATCC 23456 17 ± 0.0 22 ± 0.0 -
Shigella dysentriae ATCC 24162 30 ± 0.0 22 ± 0.0 12 ± 0.0
Klebsiella pneumonia ATCC 34089 27 ± 0.0 17 ± 0.0 7 ± 0.0
Bacillus subtilis ATCC 21332 37 ± 0.0 27 ± 0.0 10 ± 0.0

Table 1b: Antibacterial activities of the fruit of Musa paradisical Zone of inhibition (mm)

Organisms Methanol Ethanol Acetone

Escherichia coli 20 ± 0.0 - -
Escherichia coli ATCC 35218 17 ± 0.0 22 ± 0.0 -
Staphylococcus aureus 22 ± 0.0 - -
Staphylococcus aureus ATCC 25923 22 ± 0.0 9 ± 0.0 -
Salmonella typhi 27 ± 0.0 10 ± 0.0 -
Salmonella typhi ATCC 22648 27 ± 0.0 17 ± 0.0 -
Salmonella typhi ATCC 23456 22 ± 0.0 17 ± 0.0 -
Shigella dysentriae ATCC 24162 27 ± 0.0 - -
Klebsiella pneumonia ATCC 34089 7 ± 0.0 - -
Bacillus subtilis ATCC 21332 27 ± 0.0 17 ± 0.0 7 ± 0.0

Table 2a: Determination of minimum inhibition concentration (MIC) of Musa paradisica peel extract

Organisms Methanol Ethanol Acetone

Escherichia coli 150 200 _
Escherichia coli 35218 ATCC 150 200 _
Salmonella typhi 150 250 _
Salmonella typhi 22648 ATCC 150 250 _
Salmonella typhi 23456 ATCC 150 200 _
Staphylococcus aureus 200 200 _
Staphylococcus aureus 25923 ATCC 200 150 _
Shigella dysentrariae 24162 ATCC 150 200 _
Klebsiella pneumonia 34089 ATCC 200 250 _
Bacillus subtilis 21332 ATCC 100 200 _

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Table 2b: Determination of minimum inhibition concentration (MIC) of Musa paradisica Fruit extracts (mg/ml)

Organisms Methanol Ethanol Acetone

Escherichia coli 200 250 _
Escherichia coli 35218 ATCC 200 250 _
Salmonella typhi 150 300 _
Salmonella typhi 22648 ATCC 200 300 _
Salmonella typhi 23456 ATCC 200 250 _
Staphylococcus aureus 250 250 _
Staphylococcus aureus 25923 ATCC 250 200 _
Shigella dysentrariae 24162 ATCC 200 250 _
Klebsiella pneumonia 34089 ATCC 250 200 _
Bacillus subtilis 21332 ATCC 150 200 250

Table 3a: Determination of Minimum Bactericidal Concentration (MBC) of peel extract (mg/ml)

Organisms Methanol Ethanol Acetone

Escherichia coli 200 250 _
Escherichia coli35218 ATCC 200 250 _
Salmonella typhi 250 300 _
Salmonella typhi22648 ATCC 250 300 _
Salmonella typhi23456 ATCC 200 250 _
Staphylococcus aureus 200 250 _
Staphylococcus aureus 25923 ATCC 200 250 _
Shigella dysentrariae24162 ATCC 250 _ _
Klebsiella pneumonia34089 ATCC 300 _ _
Bacillus subtilis21332 ATCC 200 250 _

Table 3b: Determination of minimum bactericidal concentration (MIC) of fruit Extract(mg/ml)

Organisms Methanol Ethanol Acetone

Escherichia coli 300 _ _
Escherichia coli 35218 ATCC 250 250 _
Salmonella typhi 200 250 _
Salmonella typhi 22648 ATCC 200 250 _
Salmonella typhi 23456 ATCC 200 250 _
Staphylococcus aureus 250 _ _
Staphylococcus aureus 25923 ATCC 250 300 _
Shigella dysentrariae 24162 ATCC 300 _ _
Klebsiella pneumonia 34089 ATCC 250 300 _
Bacillus subtilis 21332 ATCC 200 250 _

The highest sensitivity exhibited on the test organisms was 40 mm with methanol extract of the least inhibition was
7 mm with acetone extract. However, the highest inhibition exhibited by commercial antibiotic on test solates was
30 mm by gentamicin and ofloxacin and least inhibition of 7 mm by cefuroxime.

The minimum inhibitory concentration (MIC) of the extracts ranged from 50 – 300 mg/ml as shown in Table 2a and
b. It was observed that at higher concentration there was a stronger activity against test organisms. There was no
activity at lower concentration against the test isolates. The result obtained ascertained 100-300 mg/ml as MIC value
for plantain peel and 200-300 mg/ml for plantain fruit extracts. The MIC value for plantain peel methanol extract on
Escherichia coli, Salmonella typhi, Staphylococcus aureus (clinical and typed isolates) and Bacillus subtilis (ATCC
21332) were between 100 - 200 mg/ml while the ethanol extract was valued at 200 mg/ml on each of this isolates.
The MIC value of fruit extract on Escherichia coli (clinical and typed isolates) and Shigella dysentrariae (ATCC
24162) was 200 and 250 mg/ml for methanol and ethanol extract and Bacillus subtilis (ATCC 21332) with 150 and
200 mg/ml for methanol and ethanol extract respectively.

The minimum bactericidal concentration (MBC) of the extract was evaluated between 50 – 300mg/ml as shown in
table 3a and b. The MBC value for methanol and ethanol peel extract on Escherichia coli 35218 ATCC and clinical
isolate, Staphylococcus aureus 25923 ATCC and S. aureus, Salmonella typhi 22648 ATCC and S. typhi and Bacillus
subtilis 21332 ATCC were 200 and 250mg/ml respectively, MBC value for methanol extract on Shigella
dysentrariae 24162 ATCC was 250mg/ml and Klebsiella pneumonia 34089 ATCC 300mg/ml.

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Figure 1: Antibiotic susceptibility pattern on Gram positive isolates

Figure 2: Antibiotic susceptibility pattern on Gram negative isolates

The MBC value for methanol and ethanol fruit extract on Escherichia coli 35218 ATCC and E. coli, Staphylococcus
aureus 25923 ATCC and S. aureus, Salmonella typhi 22648 ATCC and S. typhi and Bacillus subtilis 21332 ATCC
was 200 and 250mg/ml respectively, MBC value for methanol extract on Shigella dysentrariae 24162 ATCC was
300mg/ml and Klebsiella pneumonia 34089 ATCC 250 and 300mg/ml for methanol and ethanol extracts. The

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results of the antimicrobial screening of the methanol, ethanol and acetone extracts of the peels and fruits against ten
human pathogenic microbes, (bacteria) showed that methanol extract of the peel was more effective as compared to
the methanol extract of the fruit and others. Staphylococcus aureus, Escherichia coli ATCC 35218, Bacilus subtilis
ATCC 21332, Staphylococcus aureus ATCC 25923, Samonella typhi ATCC 22648, Klebsiella pneumoniae ATCC
34089, and Shigella dysentrariae ATCC 24162 among other microbes were comparatively more inhibited by both
peel and fruit methanolic extracts (Tables 1a &1b). This result shows that the extract of M. paradisiacal peels has
more antibacterial properties than the fruits. The ethanol extract of both fruit and peel exhibited higher antimicrobial
activities at all concentrations compared to the acetone extract (Tables 1a &1b). The microbes against which the
extracts were effective are pathogens already implicated in the etiologic and severity of human diseases. Thus, these
plant extract may be useful in Pharmaceutical and medical formulations.

DISCUSSION

This study was designed to evaluate the antimicrobial activity of plantain (Musa paradisiaca) peel and fruit. Both
the plantain peel and fruit extracts exhibited antibacterial potentials on Gram positive and Gram negative bacteria
most especially with methanol extract. However, the bacterial species were more susceptible to plantain peel
extracts than fruit extracts. Similar result was reported by [11]. Effects proving this might be the higher percentage
of hydrocarbon, monoterpene and oxygenated monoterpene appreciated for their antibacterial potentials in the peel
than fruit. It could also be noted that hence methanol extract exhibited higher antibacterial activity; it then signified
that methanol has the potential of extracting the antibacterial substances from the plantain samples than other
solvents. The antibacterial results obtained is similar to that reported by [12, 13]. Some literatures have reported
information on the presence of bioactive molecules in many plants, which have served as food and medicine in
health care man. Since the event of this scientific research on such discovery has been till date. The ideal about such
research is to find lasting solutions to replacing synthetic antibiotics with naturally available phytochemicals present
in plants for their low toxicity, low cost and readily available for human employment in disease treatment. [14] has
reported ethanolic and aqueous extract of unripe M. sapientum fruit. In this study similar result was obtained with M.
parasidiaca peel and fruit extracts. [15] has reported on the antibacterial activity of M. sapientum on some
pathogenic bacteria.

Higher antibacterial effects than known synthetic antibiotics were exhibited on test bacterial species. The methanol
extract of both peel and fruit had higher inhibition value on test bacteria than ethanol and acetone extracts. The
microbes against which the extracts were effective are pathogens already implicated in the etiologic and severity of
human diseases. Thus, the plant extract may be useful in antibacterial application. As a natural health product, M.
parasidiaca preparations as food may be accepted more readily than prescription drugs for some patient groups,
particularly in some communities afflicted with varying incidence of bacterial diseases and a paucity of culturally
acceptable treatment options.

This result showed that M. parasidiaca though taken as food for carbohydrate source could serve as agent of
bacterial inhibition.

REFERENCES

[1] Kaur Sarabjot; Monda Poonam. Journal of Microbiology and Experimentation, 2014, 1(1), 1 – 6
[2] BC. Aridoˇgan H. Baydar; S. Kaya; M. Demirci; A. Ozbas¸ar; Mumcu E. Archives of Pharmacal Research;
2002, 25( 6), 860–864
[3] R. Stojanovic; A. Belscak-Cvitanovic; V. Manojlovic; D. Komes; V. Nedovic; B. Bugarski. Journal of the
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[4] J. Gou; Y. Zou; J. Ahn. Food Science and Biotechnology, 2011, 20(1), 283–287.
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[12] I Ahmad; AZ. Beg.. J. Ethnopharmacol; 2001, 74, 113–123.
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[14] JF. Fagbemi; E. Ugoji; T. Adenipekun; O. Adelowotan. Afr J Biotechnol, 2009, 8(7), 1176- 1182.
[15] KS, Repon; A. Srijan; HS. Syed Sohidu;l; R. Priyanka. Asian Pacific Journal of Tropical Biomedicine; 2013,
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