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© 2016 Ewemen Resources Limited / EJMR. All rights reserved
2016| Volume 2 | Issue 2 |p. 22 - 29
Ewemen Journal of Microbial Research ISSN: 2488-9148
Available online at http://ewemen.com/category/ejmr/
Full Length Research
EVALUATION OF THE PHYTOCHEMICAL CONSTITUENTS AND ANTIMICROBIAL
ACTIVITY OF THE METHANOL AND HEXANE EXTRACTS OF CARICA PAPAYA
(CARICACEAE) ROOT
*1EMOKPAE L. A., 2EGHAFONA N. O., 1OGEFERE H. O., 3UWUMARONGIE
H. O.
1Department of Medical Laboratory Science, School of Basic
Medical Sciences, University of Benin, Benin City, Edo State,
Nigeria. 2Department of Microbiology, Faculty of Life Science,
University of Benin, Benin City, Edo State, Nigeria.
3Department of Pharmacognosy, Faculty of Pharmacy, University of
Benin, Benin City, Edo State, Nigeria.
ABSTRACT
Received 11 September, 2016 Revised 20 September, 2016 Accepted
26 September, 2016 *Corresponding Author’s Email:
[email protected]
The increase in the development of multi-drug resistance by
infectious agents and the existing knowledge of plants being a good
source of drugs motivated this study. Phytochemical screening was
carried out using standard methods of analysis while the in-vitro
antimicrobial tests were done using the agar diffusion method. The
results of the phytochemical tests revealed the presence of
steroids, flavonoids and alkaloids in both extracts while reducing
sugars, carbohydrates, saponins and tannins were present in the
methanol extract only. The antimicrobial activity result showed
that the hexane extract was more effective against the clinical
bacteria isolates while the methanol extract was more effective
against the fungi isolates used in this study. The extracts of C.
papaya could be used for the treatment of infections caused by the
susceptible microorganisms. Also, the phytoconstituents present in
each of the extract could be isolated and used as ingredients for
the formulation of drugs by pharmaceutical industries against the
susceptible organisms. Keywords: Carica papaya, Root extracts,
Methanol, Hexane, Antimicrobial, Bacteria, Fungi.
INTRODUCTION
Multi-drug resistant infectious agents are a major cause of
deaths occurring daily (Ahmed and Beg, 2001). Thereby posing a
global challenge on researchers to discover newer antimicrobials
that will be effective against them (Latha and Kannabiran, 2006).
This has led to the study of natural plants being a good source of
drugs (Pretorious and Watt, 2001) and are already being used for
the treatment of ailments in Asia and Africa (Bibitha et al.,
2002). Carica papaya is called “Pawpaw” in English and has various
parts such as seeds, root, stem, leaves, fruits and
flowers which are good sources of nutrients. They are also
useful medicinally (Fajimi et al., 2001; Krishna et al., 2008;
Senthilkumaran and Shalini, 2014). It has been reported to contain
phytochemicals, useful minerals, vitamins (Romasi et al., 2011) and
enzymes of great industrial value (Villegas, 1997). The seeds of C.
papaya was found to possess a broad spectrum antibacterial activity
(Orhue and Momoh, 2013; Peter et al., 2014; Al-Judaibi, 2015; Tariq
et al., 2015). The unripe and ripe fruit parts had antibacterial
activity (Akujobi et al., 2010; Orhue and Momoh, 2013). The leaf
extracts had antibacterial activity (Anibijuwon and Udeze,
2009;
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Alabi et al., 2012; Baskaran et al., 2012; Orhue and Momoh,
2013; Nirosha and Mangalanayaki, 2013; Peter et al., 2014; Sumathi
and Gowthami, 2014; Pandy et al., 2015; Vijayakumar et al., 2015)
and antifungal activity (Baskaran et al., 2012; Sherwani et al.,
2013; Vijayakumar et al., 2015). The stem extract had antibacterial
activity (Sumathi and Gowthami, 2014) while the root extracts
(methanol, aqueous, ethanol, ethylacetate and acetone) obtained via
hot method of extraction using Soxhlet apparatus was shown to
possess antibacterial (Doughari et al., 2007; Adejuwon et al.,
2011; Nirosha and Mangalanayaki, 2013) and antifungal activities
(Adejuwon et al., 2011). The ethanol root extract obtained via cold
method of extraction had activity against S. aureus, E. coli, S.
typhi and P. aeruginosa (Rubaka et al., 2014; Sumathi and Gowthami,
2014); with no activity against C. albicans (Sumathi and Gowthami,
2014). As a result of the quest for useful bioactives and medicinal
plant resources, after an extensive literature review, this study
was therefore designed to determine the phytochemical constituents
of the methanol and hexane extracts (obtained via cold method of
extraction) of the root of C. papaya collected from Benin City, Edo
State; as well as the antimicrobial activity of the extracts
against clinical isolates, so as to determine the nature of the
constituents (polar or non – polar) responsible for such
activities. MATERIALS AND METHODS Materials
All solvents were of the Analar grade. They were obtained from
JHD, Guandgua Chemical Ltd, China. Microbiological media were
obtained from BIOTEC, India and they include MacConkey agar, Blood
agar base, Mannitol salt agar, Mueller Hinton agar, Sabouraud
Dextrose agar, Nutrient broth and Peptone broth. Plant
Collection
The roots of mature cultivated C. papaya were harvested locally
from Ovia North - East Local Government Area of Edo State, Nigeria.
They were authenticated by a plant curator (Mr. Sunny Nweke) in the
Department of Pharmacognosy, Faculty of Pharmacy, University of
Benin, Benin city. The authenticated matured fresh root samples
were washed thoroughly 2 - 3 times with running tap water and
finally with sterile distilled water. The roots were chopped into
tiny bits to expose a large surface area and dried using hot air
oven at 40oC for 1 week. The dried root was then pulverized to fine
powder using Kenwood plant milling machine and stored in air tight
bottles till needed for analysis.
Extraction Process
The crude extract from the root of C. papaya was obtained
according to the method described by Alabi et al (2012). 10 g each
of the pulverized root of C. papaya was suspended in 250 mL of 95%
methanol and 250 mL of 95% hexane respectively. The mixtures were
allowed to stand for 72 hr, with occasional stirring at intervals
(every 24 hr) to ensure thorough extraction. The resulting extracts
were filtered through a Whatman filter paper (No. 1). The filtrates
were concentrated using rotary evaporator at a temperature of 40ºC.
The concentrate was then transferred into evaporating dishes and
placed in an oven maintained at 20oC to achieve complete dryness.
The dried extracts were weighed, kept in amber coloured bottles and
then preserved in a refrigerator prior to use. Phytochemical
tests
Phytochemical tests were carried out on the methanol and hexane
extracts of the root of C. papaya employing standard phytochemical
test procedures, to detect the presence/absence of secondary plant
metabolites such as glycosides, saponins, anthracene derivatives,
cyanogenetic glycosides, steroids, tannins, flavonoids and
alkaloids (Harborne, 1998). Microorganism isolates
The Eschericia coli, Pseudomonas aeruginosa, Staphylococcus
aureus, Candida albicans and Trychophyton tonsurans used in this
study were all clinical isolates obtained from the University of
Benin Teaching Hospital, Benin City, Nigeria. Pure isolates were
obtained by sub-culturing unto various respective selective media.
They were re-identified using standard procedures. Clinical
isolates authentication
The clinical isolates were re-identified using standard
procedures (Cowan and Steel, 1974) based on colonial morphology,
microscopy by wet mount, Gram’s staining technique and biochemical
tests. The S. aureus was inoculated on Mannitol salt agar, E. coli
and Ps. aeruginosa on MacConkey agar, while the C. albicans and T.
tonsurans on Sabouraud Dextrose agar. The cultures on Mannitol salt
agar, MacConkey and Sabouraud Dextrose agar with the C. albicans
were incubated for 24 hr at 37oC while the Sabouraud Dextrose agar
with the T. tonsurans was incubated for 4 days at 30oC. Thereafter,
wet mounts for motility and
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the presence of budding yeast cells were done on the colonies
grown. Also, smears were made and stained by Gram’s technique.
Catalase and Coagulase tests were done on the S. aureus.
Biochemical tests like sugar fermentation test, Citrate utilization
test, Urease test, Indole test and Oxidase test were done on all
the Gram negative bacilli. A needle mount was done on the T.
tonsurans. The re-confirmed isolates were grown on agar slants.
Before use, they were confirmed to be pure. Test for Antimicrobial
Activity
The antimicrobial activity of the methanol and hexane extracts
of the root of C. papaya were determined by the cup plate agar
diffusion method (Aida et al., 2001). The broth culture of each of
the microorganism was adjusted to 0.5 McFarland turbidity standard
and inoculated evenly unto the surface of Mueller Hinton agar (for
bacteria) and Sabouraud Dextrose agar (for fungi) using a sterile
swab. Petri-dishes of 9 cm in diameter were used. A sterile cork
borer was used to bore two (2) wells of 9 mm each in diameter, on
each of the agar plates used. The hexane extract was weighed and
reconstituted in varying volumes of dimethylsulphoxide (DMSO) to
obtain different concentrations such as 25 mg/mL, 50 mg/mL, 100
mg/mL, 150 mg/mL and 200 mg/mL. The methanol extract was also
weighed and reconstituted in varying volumes of methanol to obtain
varying concentrations as stated above (25 – 200 mg/mL). Aliquots
of 1 mL of the various dilutions of the extracts were used. In each
agar plate, one of the wells was filled with a concentration of the
plant extract (test) and the other filled with 1 mL of the diluent
(DMSO or Methanol respectively), as control. The cultures were
incubated at 37oC for 24 hr (for bacteria) and 48 hr (for fungi).
The antimicrobial activity of the root extracts were determined by
subtracting the diameter (mm) of the zone of inhibition formed
around the control well from the diameter (mm) of the zone of
inhibition formed around the test well. The antibacterial activity
of the most commonly sensitive broad spectrum antibiotics (Imipenem
10 µg and Amikacin 30 µg) discs in Medical Microbiology Department
of the University of Benin Teaching Hospital, Benin-City, Nigeria
against the clinical bacteria isolates were determined using disc
diffusion technique. For the fungi isolates, the antifungal
activity of the most commonly prescribed antifungal Ketoconazole,
at varying concentrations of 25 mg/mL,50 mg/mL,100 mg/mL, 150 mg/mL
and 200
mg/mL was also determined, using the cup plate agar diffusion
method. The diluent (sterile distilled water) was included as
control. For each plant extract, five replicate trials were
conducted against each microorganism. Determination of Minimum
Inhibitory Concentration (MIC) of the extracts
The MIC’s of the methanol and hexane extracts were determined by
the agar dilution method described by Vinothkumar et al (2010). A
stock concentration of 400 mg/mL of the extracts was prepared, from
which plates containing varying concentrations (25 – 200 mg/mL) of
each extract were made. Bacterial and fungal isolates grown in
overnight broth were diluted to 108 cfu/mL corresponding to 0.5
McFarland density and 0.025 mL volume of the different isolates
were spotted on the surface of the agar plates, at marked segments
containing various concentrations of the test extract. The plates
were appropriately incubated (37oC for bacteria and 30oC for
fungi). In all cases, the lowest concentration at which there was
no observable bacterial or fungal growth was recorded as the MIC.
Determination of Minimum Bacteriocidal Concentration (MBC) and
Minimum Fungicidal Concentration (MFC) of the extracts
The plates with no visible growth recorded for MIC determination
were swabbed with sterile swab sticks and inoculated on fresh
Nutrient agar and Sabouraud Dextrose agar plates without the
extracts. All plates were appropriately incubated. The MBC was the
lowest concentration of extracts that showed no observable growth
of test organisms while the MFC was defined as the lowest
concentration of the extracts that showed no visible growth of the
test fungus. RESULTS AND DISCUSSION
The phytochemical tests showed the presence of reducing sugars,
carbohydrates, saponins, steroids, tannins, flavonoids and
alkaloids in the methanol extract, while steroids, flavonoids and
alkaloids were present in the hexane extract. Anthracene
derivatives and cyanogenetic glycosides were absent in the methanol
extract while reducing sugars, carbohydrates, saponins, tannins,
anthracene derivatives and cyanogenetic glycosides were absent in
the hexane extract (Table 1). The phytoconstituents found in the
methanol extract in this study are same as that reported by
Doughari et al.,
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2007. The presence of bio-active substances in plants has been
reported to develop resistance against bacteria, fungi and pests on
plants (Srinivassan et al., 2001). This may explain the
antimicrobial activity exhibited by the root extract of C. papaya
used in this study. Table 1: Phytochemical constituents of the
methanol and hexane extracts of C. papaya root
Constituents Methanol extracts Hexane extracts Reducing sugars +
- Carbohydrates + - Saponins + - Anthracene derivatives - -
Cyanogenetic glycosides - - Steroids + + Tannins + - Flavonoids + +
Alkaloids + + Key: + means Present and - means Absent.
Carbohydrates consist mainly of carbon, hydrogen and oxygen with
the last two elements usually present in a ratio of 2:1. They are
the first product of photosynthesis. Sugars unite with a variety of
compounds to form glycosides. Saponins are well known for their
detergent and haemolytic properties. They are highly toxic when
injected into the blood stream but non – toxic when ingested by
mouth. They are glycosides hydrolyzed by acids to give the sugar
portion (glycone) and non – sugar portion (aglycone or sapogenin).
Tannins usually combine with proteins in animal hides, converting
them into leather by preventing their putrefaction. They may be
true tannins (hydrolysable, condensed or complex tannins)
having molecular weights of 1000 – 5000 or pseudotannins which
are of relatively lower weights. Plants containing steroidal
materials (e.g. hecogenin from Agave sisalena) are useful as
starting materials for synthesis of hormones and contraceptives.
Alkaloids are basic in nature due to one or more nitrogen atoms
usually present in a heterocyclic ring and produce a marked
physiological action on both men and animals (Evans, 2002). The
presence of these phytochemicals may be responsible for the
antimicrobial activity observed with the plant extracts, as some of
them e.g. tannins and alkaloids have been reported to have
antimicrobial activities. The clinical isolates used in this study
showed varying degree of sensitivity to the various concentrations
of the methanol and hexane extracts of C. papaya root, with the
antimicrobial activity being concentration dependent. The results
of the antimicrobial activities of the methanol extract revealed
that, at 25 mg/mL concentration, there was no antimicrobial
activity against any of the clinical isolates used. At 50 mg/mL,
there was activity against E. coli and Ps. aeruginosa. At higher
concentrations, antimicrobial activity was observed against all the
clinical isolates used. The activity was highest against E. coli at
a concentration of 200 mg/mL, producing a zone of inhibition of
12.80 mm and least against C. albicans and T. tonsurans at a
concentration of 100 mg/mL, with a zone of inhibition of 0.40 mm
(Table 2).
Table 2: Antimicrobial activity of the methanol extract of C.
papaya root
Microorganisms Concentrations (mg/mL)/Zone of Inhibitions (mm).
25 mg/mL 50 mg/mL 100 mg/mL 150 mg/mL 200 mg/mL
E. coli 0.00 ± 0.00 1.20 ± 0.49 4.80 ± 0.49 7.40 ± 0.40 12.80 ±
0.49 Ps. aeruginosa 0.00 ± 0.00 3.80 ± 0.66 7.80 ± 0.20 9.20 ± 0.49
12.00 ± 0.63 S. aureus 0.00 ± 0.00 0.00 ± 0.00 3.20 ± 0.49 5.00 ±
0.45 7.80 ± 0.20 C. albicans 0.00 ± 0.00 0.00 ± 0.00 0.40 ± 0.40
3.60 ± 1.17 7.60 ± 0.75 T. tonsurans 0.00 ± 0.00 0.00 ± 0.00 0.40 ±
0.40 2.00 ± 0.00 5.80 ± 0.66 Key: Values are Mean ± SEM. n = 5.
Table 3: Antimicrobial activity of the hexane extract of C.
papaya root
Microorganisms Concentrations (mg/mL)/Zones of Inhibition
(mm)
25 mg/mL 50 mg/mL 100 mg/mL 150 mg/mL 200 mg/mL E. coli
0.00±0.00 2.40±0.75 6.00±0.89 12.40±0.24 16.40±0.40 Ps. aeruginosa
0.00±0.00 1.80±0.49 4.20±0.20 7.40±0.24 12.00±0.00 S. aureus
0.00±0.00 1.60±0.40 4.00±0.63 9.00±0.32 12.60±0.40 C. albicans
0.00±0.00 0.00±0.00 0.00±0.00 1.40±0.24 3.00±0.45 T. tonsurans
0.00±0.00 0.00±0.00 0.00±0.00 2.20±0.20 5.20±0.49 Key: Values are
Mean ± SEM. n = 5.
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The hexane extract had no antimicrobial activity against any of
the clinical isolates used for this study at a concentration of 25
mg/mL. At 50 and 100 mg/mL, it had activity against all the
bacteria isolates but none against any of the fungi isolates used.
However, antimicrobial activity increased with the concentration of
the extract, with activity observed against all the clinical
isolates at concentrations of 150 and 200 mg/mL. The activity was
highest against E. coli at a concentration of 200 mg/mL, which
produced a zone of inhibition of 16.40 mm and least against C.
albicans at a concentration of 150 mg/mL, with zone of inhibition
of 1.40 mm (Table 3). When a comparison of the antimicrobial
activities of the methanol and hexane extracts of the root of
C.
papaya on all the clinical isolates used in this study was done,
it was observed that both extracts had no activity at 25 mg/mL
concentration against any of the clinical isolates. At 50 mg/mL,
methanol extract had activity against E. coli and P. aeruginosa
whereas hexane extract had activity against all the bacteria
isolates. At 100 mg/mL, methanol extract had activity against all
the clinical isolates used unlike hexane extract which had activity
against only the bacteria isolates. At 150 and 200 mg/mL
concentrations, both extracts had activity against all the clinical
isolates used to varying degrees (Figure 1). Hence, against the
bacteria isolates, hexane extract was more effective than the
methanol extract and viz versa for the fungi isolates. Also, at 100
mg/mL concentration, the methanol had a broader spectrum of
activity than the hexane extract.
Figure 1: Chart of antimicrobial activities of the methanol and
hexane extracts of C. papaya root.
Figure 2: Chart of antifungal activities of the methanol and
hexane extracts of C. papaya root with Ketoconazole.
0
2
4
6
8
10
12
14
16
18
50 mg/mL 50 mg/mL 100 mg/mL 100 mg/mL 150 mg/mL 150 mg/mL 200
mg/mL 200 mg/mL
Methanol Hexane Methanol Hexane Methanol Hexane Methanol
Hexane
Mea
n v
alu
e o
f Zo
ne
of
Inh
ibit
ion
(m
m)
Escherichia coli Staphylococcus aureusPseudomonas aeruginosa
Candida albicansTrichophyton tonsurans
0
1
2
3
4
5
6
7
8
9
25 mg/mL 50 mg/mL 100 mg/mL 100 mg/mL 150 mg/mL 150 mg/mL 150
mg/mL 200 mg/mL 200 mg/mL 200 mg/mL
Ketoconazole Ketoconazole Methanol Ketoconazole Methanol Hexane
Ketoconazole Methanol Hexane Ketoconazole
Mea
n v
alu
e o
f Zo
ne
of
Inh
ibit
ion
(m
m)
Candida albicans Trichophyton tonsurans
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The antibacterial activity of the reference antibiotics
(Imipenem and Amikacin) in discs, with very low concentrations of
10 µg and 30 µg respectively, exhibited good antibacterial
activities. Imipenem had greater activity against E. coli and S.
aureus than Amikacin, which had a greater activity against the Ps.
aeruginosa used (Table 4). This good antibacterial activity of the
reference antibiotics can be attributed to the fact that, the
active drugs are in the pure form. This is unlike the methanol and
hexane extracts of the plant material which are in the crude form,
with many ingredients in them, possibly synergizing or antagonizing
the activity of each other. Hence, the reference drugs had better
antibacterial activities against the clinical bacteria isolates
than the extracts used in this study. However, the extracts had a
broader spectrum of activity against the clinical isolates when
compared to the reference antibiotics. This is because the extracts
(methanol and hexane), both have activities against Gram positive
and Gram negative bacteria, as well as the fungi isolates used in
this study.
Table 4: Antibacterial activity of reference antibiotic discs
against the bacterial isolates
Microorganisms Zones of Inhibition (mm)
Imipenem disc (10µg)
Amikacin disc (30µg)
E. coli 1.92±0.05 1.68±0.04 Ps. aeruginosa 1.20±0.03 1.68±0.04
S. aureus 3.28±0.08 1.78±0.02 Key: Values are Mean ± SEM, n =
5.
The antifungal activity of the reference antifungal Ketoconazole
at varying concentrations similar to that of the extracts, had
antifungal activity against the fungi isolates used (C. albicans
and T. tonsurans). These organisms were more sensitive to
Ketoconazole than the plant extracts, as a concentration of 25
mg/mL produced zones of inhibition of 2.94 and 2.32 mm against C.
albicans and T. tonsurans respectively (Table 5).
Table 5: Antifungal activity of Ketoconazole against the fungi
isolates
Microorganisms Zones of inhibition (mm)
25 mg/mL 50 mg/mL 100 mg/mL 150 mg/mL 200 mg/mL C. albicans
2.94±0.04 3.72±0.05 4.24±0.07 5.36±0.04 6.48±0.05 T. tonsurans
2.32±0.05 3.18±0.05 3.50±0.03 4.02±0.02 4.92±0.05 Key: Values are
Mean ± SEM, n = 5.
Despite these fungi isolates being more sensitive to
Ketoconazole at concentrations of 25 – 150 mg/mL, at 200 mg/mL
concentration, the methanol extract of the plant material was
observed to have better activity than Ketoconazole. This is unlike
the hexane extract which only had a better activity against T.
tonsurans. Hence, for children and adults (>30 Kg) with fungal
infections whose recommended dose is 200 mg once daily or twice
daily, the methanol extract would be more effective against
infections caused by these susceptible organisms (Figure 2). This
is unlike for patients
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account for variations in activities of same plant (or part)
collected from different locations. For example, the MIC of the
methanol extract against Ps. aeruginosa in this study was found to
be 50 mg/mL which contracts that of Doughari et al., (2007) found
to be 100 mg/mL. This difference in activity may have resulted from
the different locations (Benin City, Edo State and Yola, Adamawa
State; respectively) where the plants were collected, as method of
extraction (cold or hot) used in the studies had no significant
effect on the activities of the root extracts against E. coli and
S. aureus. Also, these various phytoconstituents yield themselves
to extraction based on the polarity of the solvents. Polar solvents
such as water and methanol tend to extract more of the polar
phytoconstituents in plants while non-polar solvents such as hexane
and petroleum ether tend to extract the non-polar
phytoconstituents. Hence, from this study, it could be said that,
the non-polar constituents in the hexane extract of C. papaya root
are more effective against the bacterial isolates. This is unlike
results gotten from the studies of Nirosha and Mangalanayaki, 2013
and Rubaka et al., 2014 were the more polar ethanol extract had
better antibacterial activity than the less polar ethyl acetate and
petroleum ether extracts respectively. Also, from this study, the
polar constituents in the methanol extract were found to be more
effective against the fungi isolates. This is similar to results
obtained by Adejuwon et al., 2011 were polar solvents (methanol and
water) had good activity against Trichophyton rubrum,
Epidermophyton floccosum and Microsporum audouinii. However,
results obtained in this study contradicts that obtained by Sumathi
and Gowthami, 2014 were polar solvents (ethanol and water) had no
activity against C. albicans. CONCLUSION
The extracts (methanol and hexane) have been shown to contain
useful phytochemicals which are usually responsible for the
biological activities observed in plants, when used for the
treatment of various ailments. These useful phytochemicals could be
isolated and used as ingredients in the formulation of drugs by
pharmaceutical industries. Also, the extracts showed antimicrobial
activities at the tested concentrations against the selected
clinical isolates used in this study. Hence, they could be used for
the treatment of infections such as bacteremia and urinary tract
infections, sepsis, yeast infections and Tinea capitis caused by E.
coli and Ps. aeruginosa, S. aureus, C. albicans and T. tonsurans
respectively.
ACKNOWLEDGEMENT
The technical help of Members of Staff, Department of Medical
Microbiology of the University of Benin Teaching Hospital, Benin
City, Edo State, Nigeria, is appreciated and thankfully
acknowledged. CONFLICT OF INTEREST
None declared. REFERENCES
1. Adejuwon AO, Agbaje EO and Idika N (2011). Antifungal and
antibacterial activities of aqueous and methanolic root extracts of
Carica papaya Linn. (Caricaceae). Int Res J Micro 2 (8): 270 –
277.
2. Ahmad I and Beg AZ (2001). Antimicrobial and Phytochemical
studies on 45 Indian medicinal plants against multi-drug resistant
human pathogens. J Ethnopharmacol 74: 87 - 91.
3. Aida P, Rosa V, Blamea F, Tomas A and Salvador C (2001).
Paraguyan plants used in traditional medicine. Short communicaton.
J Ethnopharmacol 16: 93 - 98.
4. Akujobi CN, Ofodeme CN and Enweani CA (2010). Determination
of antibacterial activity of Carica papaya (Pawpaw) extracts. Nig J
Clin Pract 13 (1): 55 - 57.
5. Alabi OA, Haruna MT, Anokwuru CP, Jegede T, Abia H, Okegbe VU
and Esan BE (2012).Comparative studies on antimicrobial properties
of extracts of fresh and dried leaves of Carica papaya (L) on
clinical bacterial and fungal isolates. Advan Appli Sci Res 3 (5):
3107 - 3114.
6. Al-Judaibi A (2015). Comparative study of plant extracts as
broad-spectrum antibacterial agents. Int J Eng Sci 5 (7): 27 -
33.
7. Anibijuwon II and Udeze AO (2009). Antimicrobial Activity of
Carica Papaya (Pawpaw Leaf) on some pathogenic organisms of
clinical origin from South-Western Nigeria. Ethnobotan Leaflets 13:
850 - 864.
8. Baskaran C, Ratha bai V, Velu S and Kumaran K (2012). The
efficacy of Carica papaya leaf extract on some bacterial and a
fungal strain by well diffusion method. Asian Pac J Trop Disease,
S658 - S662.
9. Bibitha B, Jisha VK, Salitha CV, Mohan S and Valsa AK.
(2002). Antibacterial activity of different plant extracts. Short
Communication. Indian J Microbiol 42: 361 - 363.
10. Cowan ST and Steel KJ (1974). Manual for the identification
of medical bacteria. Cambridge University Press. pp. 945.
11. Doughari JH, Elmahmood AM and Manzara S (2007). Studies on
the antibacterial activity of root extracts of Carica papaya. L.
Afri J Microbiol Res 1: 37 - 41.
12. Evans WC (2002). Trease and Evans Pharmacognosy. 15th
edition. W. B Saunders, Philadelphia. pp. 3 – 350.
13. Fajimi AK, Taiwo AA, Ayodeji IO, Adebowale EA and Ogundola
FI (2001). Therapeutic trials on gastrointestinal helminth parasite
of goats using pawpaw seeds as a drench. Proceedings of the
International conference on sustainable crop. Livestock production
for improved livelihoods and Natural Resource Management, West
Africa. Held at the International Livestock Research Institute
(ILRI) in partnership with International Institute of Tropical
Agriculture between Nov. 9 – 23, 2001.
14. Harbone JB (1998). Phytochemical methods; A guide to modern
techniques of plant analysis. Chapman and Hills, London. pp. 30 –
297.
-
Ewemen Journal of Microbial Research 2016, 2(2): 22 - 29 Emokpae
et al.
www.ewemen.com Page 29
15. Krishna KL, Paridhavi M and Jagruti AP (2008). Review on
nutritional, medicinal and pharmacological properties of Papaya
(Carica papaya Linn.). Nat Prod Rad 7 (4): 364 - 373.
16. Latha SP and Kannabiran K (2006). Antimicrobial activity and
phytochemicals of Solanum trinobatum Linn. Afri J Biotech 5 (23):
2402 - 2404.
17. Nirosha N and Mangalanayaki R (2013). Antibacterial activity
of leaves and stem extract of Carica papaya L. Int J Advan Pharm
Biol Chem 2 (3): 473 – 476.
18. Orhue PO and Momoh ARM (2013). Antibacterial activities of
different solvent extracts of Carica papaya fruit parts on some
gram positive and gram negative organisms. Int J Herbs Pharmacol
Res 2 (4): 42 – 47.
19. Pandey B, Chandrakar V and Deshpande B (2015). Study of
antibacterial activity of crude extract of leaves of Carica papaya.
Indian J L Sci 5 (1): 033 – 036.
20. Peter JK, Kumar Y, Pandey P and Masih H (2014).
Antibacterial activity of seed and leaf extract of Carica papaya
var. Pusa dwarf Linn. J Pharm Biol Sci 9 (2): 29 – 37.
21. Pretorius CJ and Watt E (2001). Purification and
identification of active components of Carpobrotus edulis L. J
Ethnopharmacol 76: 87 - 91.
22. Romasi EF, Karina J and Parhusip AJN (2011). Antibacterial
activity of papaya leaf extracts against pathogenic bacteria.
Makara J Tech 15 (2): 173 - 177.
23. Rubaka C, Ndakidemi P, Malebo HM and Shahada F (2014).
Individual and combined antibacterial activity of crude extracts
from medicinal plants Carissa spinarum Linn and Carica papaya Linn.
European J Med Plants 4 (12): 1513 - 1523.
24. Senthilkumaran JJ and Shalini N (2014). An overview of
Carica papaya and its medicinal uses. Res J Pharm Bio Chem Sci 5
(2): 641 - 649.
25. Sherwani SK, Bokhari TZ, Nazim K, Gilani SA and Kazmi SU
(2013). Qualitative phytochemical screening and antifungi
activity of Carica papaya leaf extract against human and plant
pathogenic fungi. Int Res J Pharm 4 (7): 83 – 86.
26. Srinivasan D, Perumalsamy LP, Nathan A and Sures T (2001).
Antimicrobial activity of certain Indian medicinal plants used in
folkloric medicine. J Ethnopharmacol 94: 217 - 222
27. Sumathi R and Gowthami M (2014). Phytochemical analysis and
in- vitro antimicrobial activity of aqueous and solvent extracts of
Carica papaya against clinical pathogens. Int J Adv Res Biol Sci 1
(1): 73 - 77.
28. Tariq MH, Ghaffar B, Ahmed T, Sultan A, Irfan M and Farrukh
MJ (2015). Phytochemical and microbiological evaluation of
different chemical extracts of papaya seeds on clinical isolates of
(Fgsh Hospital) Islamabad. Int J Pharm 5 (1): 122 - 126,
29. Vijayakumar M, Bharathidasan R and Prince L (2015).
Antimicrobial activity of Carica papaya L. Int J Arts Sci Res 2
(2): 37 – 43.
30. Villegas VN (1997). Edible fruits and nuts - Carica papaya
L. EWM Verheij, volume 2. Coronel R.E. Edition, Wageningen
University, The Netherlands. pp 198.
31. Vinothkumar P, Siveraj A, Ahmed KSZ, Sivamani P, Devi K and
Senthikumar B (2010). Evaluation of antibacterial activities of
Andrographis paniculata leaf extract against Gram negative and Gram
positive species by in – vitro methods. J Pharmacol Res 3: 1513 –
1515.
Article’s Citation:
Emokpae LA, Eghafona NO, Ogefere HO and Uwumarongie HO (2016).
Evaluation of the phytochemical constituents and antimicrobial
activity of the methanol and hexane extracts of Carica papaya
(caricaceae) root. Ew J Microb Res 2(2): 22 - 29.