1. _____________________________________________________________________________________________________
*Corresponding author: Email: kurup_rajini@yahoo.com;
Original Research Article
Journal of Biology and Nature
3(3): 87-93, 2015
International Knowledge Press
www.ikpress.org
ANTIMICROBIAL PROPERTIES OF THE FRUIT PULP OF
THREE LOCAL FRUITS: Morinda citrifolia, Persea americana
AND Musa sapientum IN GUYANA
GINEL NEDD1
, RAJINI KURUP2*
, ABDULLAH ADIL ANSARI1
AND KEISHA HOLDER1
1
Faculty of Natural Sciences, University of Guyana, Guyana.
2
Faculty of Health Sciences, University of Guyana, Guyana.
AUTHORS’ CONTRIBUTIONS
This work was carried out in collaboration between all authors. Authors GN and AAA designed the study, wrote
the protocol and interpreted the data. Authors RK and GN anchored the field study, gathered the initial data and
performed preliminary data analysis. Author KH did the biochemical analysis. While authors RK and AAA
managed the literature searches and produced the initial draft. All authors read and approved the final
manuscript.
Received: 5th
May 2015
Accepted: 22nd
May 2015
Published: 17th
June 2015
__________________________________________________________________________________
ABSTRACT
The present study aim to evaluate the antimicrobial properties of Morinda citrifolia (Noni), Persea americana
(Avocado) and Musa sapientum (Banana) fruits against certain bacterial and fungal strains using disc diffusion
method. All the test extracts exhibited significant antimicrobial activity on certain pathogens. The highest zone
of inhibition occurred on Staphylococcus sp. by the ethanol avocado extract. Overall, Staphylococcus was most
susceptible to the ethanol extracts while Candida albicans was the most resistant. The extracts were proved as
strong inhibitors against bacteria than fungi, however the test extracts of Avocado found to be effective on both
bacteria and fungi. The phytochemical chemical analysis of the fruit extracts revealed the presence of common
phytochemicals such as tannins, flavonoids, alkaloids, reducing sugar, saponins and glycosides. These
phytoconstituents found in the fruit extracts would be viable in treating infectious diseases.
Keywords: Antimicrobial activity; phytochemicals; Morinda citrifolia; Persea americana; Musa sapientum;
disc diffusion.
1. INTRODUCTION
According to World Health Organization, medicinal
plants are the best source to obtain a variety of new
herbal drugs [1]. Considering the vast potentiality of
plants as sources for antimicrobial drugs with
reference to antibacterial and antifungal agents, an
investigation was undertaken to screen three local
fruits of Guyana for their antibacterial and antifungal
properties. Previous studies have shown that
medicinal plants are one of the best resources for the
isolation and development of novel bioactive
molecules [2]. Moreover, plant-derived preparations
have drawn attention of people worldwide because of
their fewer side effects and lesser toxicity in
comparison to synthetic drugs. To date, many
medicinal plants have been screened for their
therapeutic potential and their pharmacological

2. Nedd et al; JOBAN, 3(3): 87-93, 2015
88
properties like antimicrobial and antioxidant effects
[3]. Extensive search for natural antioxidants that
could minimize free radical induced damage to bio
molecules like lipids, proteins and nucleic acids is
going on worldwide. These natural antioxidants might
play an important role in combating oxidative stress
associated degenerative diseases such as cancer,
cardiovascular diseases, diabetes, atherosclerosis,
Alzheimer’s disease and aging [4]. Peels and seeds
are often the waste part of various fruits. Interestingly,
the peel and seed fractions of some fruits were found
to show higher antimicrobial and antioxidant activity
than the pulp fractions [5-8]. Different studies have
proved that the pulp of Banana has a significant
healing effect due to their antioxidant activities and
have potential antifungal and antibiotic properties
[9-11]. The phytochemical components of banana,
tannins, eugenol and tyra-mine have been proven to
have antibacterial effects [12]. Other active
compounds present in banana such as alkaloids,
glycosides, flavonoids, saponins, steroids, serotonin
and dopamine also contribute to pharmacological
effects [13].
Medicinal plants have been used by the indigenous
communities in Guyana for long time in the treatment
of various illnesses, but the science of the
effectiveness has not been explored much to provide
the scientific understanding of the concept of holistic
healing. In this series of experiment, the research was
done to prove the effectiveness of traditional medicine
using fruits with relevant causative factors
systematically explored.
2. METHODS
For the purpose of this experiment, triangulation
methodology was used. In the socio sciences,
triangulation is often used to indicate that more than
two methods are used in the study.
2.1 Fruit Selection and Collection
Banana, Avocado and Noni fruits with healthy
appearance were selected, washed with distilled
water, peeled, sliced into small pieces and shade dried
for about four weeks on stainless steel zinc with the
support of concrete blocks. Subsequently, it was
grinded into fine powder form using an electric mill
and was then stored in zip lock bags [14].
2.2 Solvent Extraction
Approximately, 10 g each of the fruit powder was
soaked in 100 ml of distilled water, hexane and
ethanol. The powder was weighed and placed into the
different solvents. Pure ethanol was used to extract
both the polar and non-polar components of the fruits;
it was poured in the conical flask with the grinded
fruits and was soaked for 48 hours. After 48 hours of
soaking the fruit powder, the solvent was decanted.
The powder was then soaked in hexane to extract non-
polar components for 48 hours was then decanted. It
was finally soaked in distilled water for 24 hours to
extract the polar components. After extraction, the
solutions obtained from the soaked fruit powder were
filtered using filter papers and subsequently stored in
air tight glass containers a room temperature. The
filtrates were vacuum dried using rotary evaporator
and concentrates were stored at 4°C [15].
2.3 Identification Test for Phytochemicals
(Secondary Metabolites)
Tests were done to detect the presence of Flavonoids,
Tannins, Reducing sugars, Alkaloids, Glycosides,
Terpenoid and Steroid. These test for the various
phytochemicals were done for both hexane and
ethanolic extracts of all the three fruits.
2.4 Flavonoid
Four millilitres (4 ml) of extract solution was treated
with 1.5 ml of 50% methanol solution. The solution
was warmed after which metal magnesium was added.
To this solution, 5 - 6 drops of concentrated
hydrochloric acid was then added, a red colour was
observed which indicated the solution was positive for
flavonoids and while an orange colour indicated the
presence of flavones [16].
2.5 Tannins
To 0.5 ml of extract solution 1 ml of water and 1 - 2
drops of ferric chloride solution was added. The
presence of gallic tannins was indicated through the
appearance of a blue colour and green black for
catecholic tannins [17].
2.6 Reducing Sugar
To 0.5 ml of extract solution, 1 ml of water and 5 - 8
drops of Fehling’s solution was added hot and
observed for brick red precipitate [18].
2.7 Alkaloid
Alkaloid solution produces white yellowish
precipitate when a few drops of Mayer’s reagents
were added [16]. The residue of the alcoholic extract
was heated on a boiling water bath with 2%
hydrochloric acid. It was allowed to cool after which
the mixture was filtered and treated with a few drops
of Mayer's reagent [19].

3. Nedd et al; JOBAN, 3(3): 87-93, 2015
89
2.8 Glycoside
To the solution of the extract, few drops of ferric
chloride and concentrated sulphuric acid were added,
and observed reddish brown coloration at the junction
of two layers and the bluish green colour in the upper
layer which was an indication of the presence of
glycoside [16].
2.9 Terpenoid and Steroid
Four milligrams of extract were treated with 0.5 ml of
acetic anhydride and 0.5 ml of chloroform,
concentrated solution of sulphuric acid were then
added slowly and red violet colour indicated a
positive test for terpenoid and green bluish colour for
steroids [16].
2.10 Test Microorganisms
The microbial strains investigated were pathogenic
organisms; Staphylococcus aureus (ATCC 25923),
Escherichia coli (ATCC 25922), Aspergillus flavus
and Candida albicans Ampicillin and Nystatin was
used as a positive control. The pathogens were
collected from Georgetown Public Hospital
Cooperation from different clinical samples. The
bacterial and fungal stock cultures were incubated for
24 hours at 37°C on nutrient agar and potato dextrose
agar (PDA) medium, respectively, following
refrigeration storage at 4°C. The bacterial strains were
grown in Mueller-Hinton agar (MHA) plates at 37°C
(the bacteria were grown in the nutrient broth at 37°C
and maintained on nutrient agar slants at 4°C),
whereas the yeasts and moulds were grown in
Sabouraud dextrose agar (SDA) and PDA media,
respectively, at 28°C. The stock cultures were
maintained at 4°C.
2.11 Antimicrobial Susceptibility Test (Disc
Diffusion Method)
Disc diffusion method was used to screen the
antimicrobial activity using NA, SDA and PDA media
plates. Concentration of the bacterial and fungal
suspensions was adjusted to 0.5 McFarland standards
(optical density) by using a spectrophotometer.
Isolates were spread on MHA plates, SDA and PDA
respectively by using sterilized cotton swabs [20-23].
The previously prepared discs of hexane and ethanolic
extract (10 µL of the extract) of Banana, Avocado and
Noni fruits were arranged radially on the agar plate.
Antibiotic discs were used as positive control and
sterilized distilled water disk was used negative
control. The zones of growth inhibition around the
disks were measured after 18 to 24 hours of in
incubation at 37°C for bacteria and 48 to 96 hours for
fungi at 28°C. The sensitivities of the microorganism
species to the extracts were determined by measuring
the sizes of inhibitory zones (including the diameter
of disk) on the agar surface around the disks, and
values <8 mm were considered as not active against
microorganisms.
2.12 Statistical Analysis
The collected data was analyzed using the following
statistical test (SPSS 20.0)
1. Mean value and standard deviation
2. One-way analysis of variance
3. RESULTS
Major Phytochemicals that were present in the fruit
extracts with different solvents are shown in Table 1.
The results indicated that only the water and noni
extract contained flavonoids while it was absent in the
others, whereas only the water and avocado extracts
contained flavones. From the results, none of solvent
extracts contained gallic tannins, alkaloids and
terpenoids.
All the assays were conducted in triplicates and
repeated thrice for consistency of results and
statistical purpose. Statistical analysis (descriptive and
comparison of means) were performed using ANOVA
for antimicrobial data to compare means of zone of
inhibition among the fruit extracts, solvents used and
the positive controls. The results from antimicrobial
assay indicated that the fruit extracts containing
ethanol, with highest antimicrobial activity Table 2.
The solvents hexane, ethanol and water alone did not
produce any zone of inhibition.
The Two-Factor without replication analysis at a P ≤
0.05 showed that the F value is less than critical F
value for hexane and water extracts; indicating there
was no significant difference on colonies of C.
albicans, A. flavus, E. coli and Staphylococcus. With
ethane extract there was significant difference on
colonies of microbes with F value > F Table 3. On the
other hand different extracts showed significant
difference on different type of microbe, which
indicates the extracts are statistically and clinically
effective in inhibiting theses microbes. Low residual
mean square demonstrates that most variation in the
data is accounted for by the separate effects of fruit
extract and type of microbe.

4. Nedd et al; JOBAN, 3(3): 87-93, 2015
90
Table 1. Major phytochemicals present in the fruit extracts containing different solvents
Solvent
extracts
Phytochemicals
Flavonoid Flavones Gallic
tannins
Catecholic
tannins
Reducing sugars Alkaloids Glycosides Steroids Terpenoid
Water &
Noni
+ - - + + - + - -
Water &
Avocado
- + - + + - + - -
Water &
Banana
- - - - + - - - -
Hexane &
Banana
- - - + - - - - -
Hexane &
Noni
- - - + - - - - -
Hexane &
Avocado
- - - - - - - + -
Ethanol &
Banana
- - - + + - + + -
Ethanol &
Noni
- - - + + - + + -
Ethanol &
Avocado
- - - + + - + + -
Table 2. The diameter of inhibition zones of three fruit extracts (Avocado, Noni and Banana) with
ethanol, methanol and aqueous solution
E. coli S. aureus C. albicans A. flavus
Ethanol & Avocado 8.75 14.75 0.33 10.25
Ethanol & Noni 11.25 13.25 0.56 11.5
Ethanol & Banana 3.25 11.75 0.53 8.5
Hexane & Avocado 0 12 0 9.25
Hexane & Noni 0 6.25 0.5 0
Hexane & Banana 0 0 0 0
Water & Avocado 0 0 0 11
Water& Banana 0 6.25 0 0
Water & Noni 0 4.75 0 13
Positive control 0 35 28.5 28.5
Table 3. The statistical analysis (ANOVA) of the
zone of inhibition seen on C. albicans, A. flavus,
E. coli, Staphylococcus when treated with solvent
extracts of Avocado, Noni and Banana
4. DISCUSSION
A number of studies have reported the potential
antibacterial activities of medicinal plant including
fruits such as banana [24]. Although some of the
antibacterial activities of medicinal plant is well-
documented, their antibacterial capacity in-vitro may
have a wide degree of variation depending on several
factors such as test medium, different methods, tested
organisms and the different in nature of the plant [25].
The phytochemical activity of Noni, Avocado and
Banana demonstrated the presences of common
phytoconstituents such as glycosides, tannins,
flavonoids, steroids, terpenoids and alkaloids, the
results are supported by Kumar and co-workers [26]
who reported that these fruits are likely to contain
these phytochemicals among others. However, not all
the phytochemicals were present when the test was
carried out. The extracts contained phytochemicals
except gallic, tannins, alkaloids and terpenoids.
In vitro studies have showed that these three fruits
Noni, Avocado and Banana showed inhibitory activity
effective against bacteria unlike fungi. Avocado,
however showed highest antibacterial effect. These
fruits are not only used for their antimicrobial effect
ANOVA: Two factor without replication
Hexane extract of 3 samples F P value F crit
Rows 1 0.5 2.8
Columns 9.9 0.0 4.8
Ethane extract of 3 samples
Rows 3.3 0.0 2.8
Columns 5.6 0.0 4.8
Water extracts of 3 samples
Rows 0.3 0.3 2.8
Columns 3.9 0.1 4.8

5. Nedd et al; JOBAN, 3(3): 87-93, 2015
91
but also for dietary purposes, inflammation, cold,
dysentery among several other disorders caused by
microbes. The results found for fruit extract was most
effective against bacteria than fungi, especially
Staphylococcus aureus. From the data it is evident
that the ethanol extracts is active against both bacteria
and fungi but more active against former. The highest
antibacterial activity was found towards S. aureus,
while it was less active against C. albicans. S. aureus,
a gram positive pyrogenic bacterium is a causative
agent towards skin diseases including superficial and
deep follicular lesion also food poisoning along with
other infections [18]. The phytochemical activity of
Noni, Avocado and Banana demonstrated the
presences of common phyto-constituents such as
glycosides, tannins, flavonoids, steroids, terpenoids
and alkaloids, the results are supported by Kumar and
co-workers [26] who reported that these fruits are
likely to contain these Phytochemicals among others.
The most critical problem related with health and
causes of mortality rates in society is infectious
disease. Since last decade, there has been an
increasing evidence of bacterial and fungal infections
due to population explosion, pollution, changed
environmental conditions, wastes from different
sources, which may affect food with perfect nutrition
value. Yadav and Agarwal investigated the qualitative
and quantitative phytochemical analysis of seven
plants of North-eastern region of India and reported
evidence of medicinally important bioactive
compounds which could be used as medicines [27]. A
similar study showed antimicrobial activity of Noni
with an increasing effect on microbial growth
inhibition with increasing concentration of the extract
[28]. Moreover, surprisingly ethanolic extract
Moreover, surprisingly ethanolic extract of Noni fruit
induce cell growth inhibition induce cell growth
inhibition on Staphylococcus aureus and Escherichia
coli and cell growth inhibition on B16-F10 cells. This
could lead to a much promising improvement in
therapeutic approach to infectious diseases and
melanoma treatment [29].
Similar to the same study, antimicrobial activity of a
defensin – an antimicrobial peptides produced by
avocado could be useful in the control of pathogens
[30]. Along with antimicrobial effect, avocado also
shares vasorelaxant effect, analgesic and anti-
inflammatory activity, hypotensive activity,
anticonvulsant effect, antiviral activity, wound
healing activity, antiulcer effect, antihepatotoxic
activity, antioxidant activity and hypoglycemic
activity [31]. Similar to other studies methanol extract
of banana extracts showed an excellent antimicrobial
property than aqueous extract [32,33]. No
antibacterial activity was shown for fruit extract with
aqueous solution in this study, which could be due to
low phenolic content extracted by the water [34].
Similar to other studies that has found water more
likely to have low selective properties as it unable
to dissolve enough bioactive compounds for
antibacterial activity [35,36].
5. CONCLUSION
The results of the present study supports folkloric
usage of the studied fruit extracts of Noni, Avocado
and Banana, all three fruit extracts possess
compounds with antimicrobial properties that need to
be explored for other viable components. It can be
safely concluded that as a result of the essential
properties that are found in the fruit extracts of Noni,
Avocado and Noni, Avocado and Banana, they are as
effective are as effective as synthetic products to
remedy infectious diseases.
COMPETING INTERESTS
Authors have declared that no competing interests
exist.
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