2016| Volume 2 | Issue 2 |p. 22 - 29 Ewemen Journal of ... · aeruginosa (Rubaka et al., 2014; Sumathi and Gowthami, 2014); with no activity against C. albicans (Sumathi and Gowthami,

© 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;

Ewemen Journal of Microbial Research 2016, 2(2): 22 - 29 Emokpae et al.

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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



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.,



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.



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.

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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



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.



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.

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2016| Volume 2 | Issue 2 |p. 22 - 29 Ewemen Journal of ... · aeruginosa (Rubaka et al., 2014; Sumathi and Gowthami, 2014); with no activity against C. albicans (Sumathi and Gowthami,