Current Trends in Biotechnology and Pharmacy Vol. 13 (4) 401-405, October 2019, ISSN 0973-8916 (Print), 2230-7303 (Online) 401 Abstract The antifungal activity of Albizia lebbeck Benth. (L.) was performed against selected pathogenic fungal strains namely Aspergillus niger, Aspergillus flavus, Penicillium citrinum and Rhizopus oryzae. Crude extracts from leaves of Albizia lebbeck Benth showed significant antimicrobial effect. Among different extracts, a methanolic extract of A. lebbeck showed highest zone of inhibition. The order of antifungal activity, expressed as minimum inhibitory concentration (MIC) of Methanol > Ethyl acetate > Petroleum ether observed for fungal strains tested. Methanol and ethyl acetate extracts exhibited significant antimicrobial activity than petroleum ether extract and may be suggested for use as natural antibiotic administration for the fungal diseases. Key words: Antifungal properties, MIC, Pathogenic fungal strains Introduction Albizia lebbeck Benth. (L.) (AL) commonly known as Shirish has number of therapeutic properties (1). AL is an economically important plant for industrial and medicinal uses. The leaves are good fodder with rich protein content (2). The plant contains saponin, macrocyclic alkaloids, phenolic glycosides and flavonols (3). In ayurvedic medicine, it is considered as an antidote against all types of poisons (4). The ayurvedic formulation of shirish like Panch shirish agada and Mahagandhahasti agad etc. has been indicated in poisoning. However, it has been established that no part of the plant has any antidotal value against either snake or scorpion venoms. In addition, the bark decoction of AL possesses anti- anaphylactic, anti-asthmatic activity and these potentials can be assumed as supportive measures in poisoning treatment (5). Saponin isolated from AL bark and methanolic pod extract of AL possess antispermatogenic effect. AL also has analgesic, anti-inflammatory, anti-diarrheal, anxiolytic and nootropic activity (6). The bark of A. lebbeck has been previously shown to possess antimicrobial activities against E. coli, S. typhi, P. aeruginosa, S. aureus, Bacillus cereus, Klebsiella aerogenes, Proteus vulgaris, Shigella boydii, Aspergillus fumigatus, Aspergillus flavus, A. niger, C. albicans, Salmonella typhimurium, Salmonella enteritidis, Shigella dysenteria, Shigella flexneri, C. albicans, Candida tropicalis and Candida kruse (7-9). Earlier investigations, a number of plants have been reported for antimicrobial properties across the world (10-12). The extract from leaves of Albizia lebbeck Benth was not reported for antifungal activities. Hence, the present study was focused on investigation of antifungal study from the leaves of Albizia lebbeck Benth. Materials and methods Plant material and preparation of crude extract : Aerial parts (leaf) of Albizia lebbeck plants were collected from botanical garden of MCAS, Rasipuram. The collected plant material was shade dried and ground well in a grinder with 2 mm diameter mesh. The dry powered plant materials (50 g) were extracted successively with 200 ml of petroleum ether, ethyl aceate, methanol Screening of Antifungal Potential of Leaf Extracts from Albizia lebbeck (L.) Benth Md. Nazneen Bobby 1 , S. Krupanidhi 1 , K. Abraham Peele1 , M. Indira 1 , E.G.Wesely 2 , A. Ranganadha Reddy 1 , T.C Venkateswarulu 1* 1, 1* Department of Biotechnology, Vignan’s Foundation for Science, Technology & Research, Vadlamudi-522213, Andhra Pradesh, India 2 Department of Botany, A.A. Govt. Arts College, Namakkal-637002, India * Corresponding Author: [email protected]Screening of Antifungal Potential of Leaf Extracts from Albizia lebbeck
Transcript
Current Trends in Biotechnology and PharmacyVol. 13 (4) 401-405, October 2019, ISSN 0973-8916 (Print), 2230-7303 (Online)
401
AbstractThe antifungal activity of Albizia lebbeck
Benth. (L.) was performed against selectedpathogenic fungal strains namely Aspergillusniger, Aspergillus flavus, Penicillium citrinum andRhizopus oryzae. Crude extracts from leaves ofAlbizia lebbeck Benth showed significantantimicrobial effect. Among different extracts, amethanolic extract of A. lebbeck showed highestzone of inhibition. The order of antifungal activity,expressed as minimum inhibitory concentration(MIC) of Methanol > Ethyl acetate > Petroleumether observed for fungal strains tested. Methanoland ethyl acetate extracts exhibited significantantimicrobial activity than petroleum ether extractand may be suggested for use as natural antibioticadministration for the fungal diseases.
known as Shirish has number of therapeuticproperties (1). AL is an economically importantplant for industrial and medicinal uses. The leavesare good fodder with rich protein content (2). Theplant contains saponin, macrocyclic alkaloids,phenolic glycosides and flavonols (3). In ayurvedicmedicine, it is considered as an antidote againstall types of poisons (4). The ayurvedic formulationof shirish like Panch shirish agada andMahagandhahasti agad etc. has been indicatedin poisoning. However, it has been establishedthat no part of the plant has any antidotal value
against either snake or scorpion venoms. Inaddition, the bark decoction of AL possesses anti-anaphylactic, anti-asthmatic activity and thesepotentials can be assumed as supportivemeasures in poisoning treatment (5). Saponinisolated from AL bark and methanolic pod extractof AL possess antispermatogenic effect. AL alsohas analgesic, anti-inflammatory, anti-diarrheal,anxiolytic and nootropic activity (6). The bark ofA. lebbeck has been previously shown to possessantimicrobial activities against E. coli, S. typhi,P. aeruginosa, S. aureus, Bacillus cereus,Klebsiella aerogenes, Proteus vulgaris, Shigellaboydii, Aspergillus fumigatus, Aspergillus flavus,A. niger, C. albicans, Salmonella typhimurium,Salmonella enteritidis, Shigella dysenteria,Shigella flexneri, C. albicans, Candida tropicalisand Candida kruse (7-9). Earlier investigations, anumber of plants have been reported forantimicrobial properties across the world (10-12).The extract from leaves of Albizia lebbeck Benthwas not reported for antifungal activities. Hence,the present study was focused on investigation ofantifungal study from the leaves of Albizia lebbeckBenth.
Materials and methodsPlant material and preparation of crudeextract : Aerial parts (leaf) of Albizia lebbeckplants were collected from botanical garden ofMCAS, Rasipuram. The collected plant materialwas shade dried and ground well in a grinder with2 mm diameter mesh. The dry powered plantmaterials (50 g) were extracted successively with200 ml of petroleum ether, ethyl aceate, methanol
Screening of Antifungal Potential of Leaf Extracts fromAlbizia lebbeck (L.) Benth
Md. Nazneen Bobby1 , S. Krupanidhi1 , K. Abraham Peele1 , M. Indira1 , E.G.Wesely2 ,A. Ranganadha Reddy1, T.C Venkateswarulu1*
1, 1*Department of Biotechnology, Vignan’s Foundation for Science, Technology & Research,Vadlamudi-522213, Andhra Pradesh, India
2Department of Botany, A.A. Govt. Arts College, Namakkal-637002, India* Corresponding Author: [email protected]
Screening of Antifungal Potential of Leaf Extracts from Albizia lebbeck
Current Trends in Biotechnology and PharmacyVol. 13 (4) 401-405, October 2019, ISSN 0973-8916 (Print), 2230-7303 (Online)
402
by using soxhlet apparatus for 48 hrs at atemperature not exceeding the boiling point of thesolvent. The aqueous extracts were filtered usingwhatman filter paper (No.1) and then concentratedin vacuums at 40 ºC using rotary evaporator. Theresidues obtained were stored in a freezer -80 oCuntil further tests (13, 14).
Agar disc diffusion method : The crude extractswere used for bioassay against the fungal specieswhich includes, Aspergillus niger (IMI no: 500308),Aspergillus flavus (500309), Penicillium citrinum(500310) and Rhizopus oryzae (500312).Inoculums were prepared from the 24 hours oldculture of standard fungal isolates in Potatodextrose broth. Potato dextrose plates wereprepared and the inocula were seeded by spreadplate method. In the prepared Potato dextroseagar plates the well was prepared with equaldistance in the size of 4mm.The prepared wellswere loaded with 250, 500, 1000 and 2000 μg ofplant extracts. The plates were incubated at 37°C for 24 - 48 h (15). Antifungal activity wasevaluated by measuring the inhibition zone inmillimeter in diameter and tabulated. All thesamples were done in triplicate. Both positive andnegative controls were determined, for negativecontrol the three solvents (petroleum ether, ethyl
acetate and methanol) were also used todetermine their effect on test organisms.
Results and discussionPlants and their preparations have been
used as medicines against infectious diseases.The plants are rich in secondary metabolites whichinclude alkaloids, glycosides, flavonoids,insecticides, steroids, related active metaboliteswhich are of great medicinal value and have beenextensively used in the drug and pharmaceuticalindustry (16). The antifungal study results revealedthat methanolic extracts of A. lebbeck conferredwidest spectrum activities that inhibited the growthof all studied pathogens with zone of inhibition.The methanolic extracts of A. lebbeck illustratedthe highest zone of inhibition against the plantpathogens Aspergillus niger (17 mm), Aspergillusflavus (18 mm), Penicillium citrinum (16 mm), andRhizopus oryzae (18 mm). The ethyl acetateextracts demonstrated maximum zone of inhibitionagainst Aspergillus niger (18 mm), Aspergillusflavus (16 mm), Penicillium citrinum (18 mm) andRhizopus oryzae (18 mm). The petroleum etherextracts demonstrated maximum zone of inhibitionagainst Aspergillus niger (19 mm), Aspergillusflavus (17 mm), Penicillium citrinum (15 mm) andRhizopus oryzae (16 mm). (Table 1).
Table 1 Anti-fungal activity of leaf extracts of Albizia lebbeck Benth against the fungal pathogens
The results of the present study confirm the antifungal activity of A. lebbeck and the antifungaleffects of leaf extracts were shown in the Figure 1 & 2.
Md. Nazneen Bobby et al
Current Trends in Biotechnology and PharmacyVol. 13 (4) 401-405, October 2019, ISSN 0973-8916 (Print), 2230-7303 (Online)
403
In previous studies, Shahid and Firdous in2012 tested the order of antifungal activity,observed in A. lebbeck benth. was seed> pod>flower > roots against six fungal strains such asAspergilus parasiticus, Aspergilus Niger, Candidaalbicans, Aspergillus effusus, Fusarium solaniand Saccharomyces cerevisiae and compared withItraconazole and AmphoteracinB. Similarly,Mohammed Nazneen Bobby et al (2012) reportedantibacterial effect of Albizia lebbeck against thebacterial pathogens and stated that methanol andethyl acetate extracts exhibited slightly higherefficacy than petroleum ether extracts. However,the antifungal activity from leaf extracts of A.lebbeck was not reported and hence, the presentstudy provides desirable data which is used informulation of therapeutic products from plantorigin. Johnson et al (2010) have reported thatmethanol was the most effective solvent for plantextraction than hexane and water (17). Uzama
Danlami and Envuladu Patience Elisha (2017)reported that the Albizia lebbeck and its mistletoeleaves extracts have good potency forantimicrobial activity and found greater zone ofinhibition against the fungal strains such asAspergillus fumigatus (16 mm), Aspergillus niger(14 mm), Fusarium oxysforum (9 mm) from theethanol extract of mistletoe leaves of Albizialebbeck (18). Similarly, Ali et al (2018) reportedthat the petroleum ether, ethyl acetate andmethanol bark extract of A. Lebbeck have shownthe significant antimicrobial activity against thetest organisms namely, Candidaarrizae, Aspergillus fumigatus, Aspergillus
Figure 2 Antifungal activity of Albizia lebbeckBenth against A.niger and A.flavus
Figure 1 Antifungal activity of Albizia lebbeckBenth against P.citrinum and R.oryzae
Screening of Antifungal Potential of Leaf Extracts from Albizia lebbeck
Current Trends in Biotechnology and PharmacyVol. 13 (4) 401-405, October 2019, ISSN 0973-8916 (Print), 2230-7303 (Online)
404
niger, Rhizopus oryzae, Candida albicans, Saccharomyces cerevisiae, Candida krusei andthey found that among all the ethyl acetate extracts of A. Lebbeck have the mostpotential antimicrobial (19). However, the findingsof the present study proved that the leaf extract ofA. lebbeck can be used as potential sourceantifungal studies.
ConclusionA. lebbeck is the potential source of
bioactive compounds that could be used toformulate potent antimicrobial drugs of naturalorigin. The findings of this study proved that theextracts of A. lebbeck has antifungal activity andwhich might be helpful in preventing the diseasescaused by pathogenic organisms and can be usedas alterative system of medicine.
AcknowledgementThe authors gratefully acknowledge the
Arignar anna government arts college, Namakkal-637002, Tamil Nadu, India and VFSTR (Deemedto be University), Guntur-522213, India for providingthe facilities to carry out this work.
Conflict of interest None declaredReferences1. Priyanka Jaswas and Padma kumar
Evaluation of Antimicrobial efficacy of etractsof a semi arid plant Albizia lebbeck (L.)Benth. Indian journal of Natural products andResources. 2016; 7(4): 287-292.
2. Sharma GK and Dubey N. Review of Shirish(Albizia lebbeck) therapeutic properties.International journal of ayurvedic and herbalmedicine 2015; 5(1): 1683–1688.
4. Gajendra kumar Sharma and Nishtha Dubey.Review of Shirish (Albizia lebbeck)therapeutic properties, International Journalof Ayurvedic and Herbal Medicine. 2015;5(1): 1683–1688.
5. Mishra SS, Gothecha VK and Sharma.Albizia lebbeck: A short review. Journal ofherbal medicine and toxicology 2010; 4 (2):9-15.
6. Misra LN, Dixit AK, Wagner H. N-demethylbudmunchiamines from Albizia lebbeckseeds. Phytochemistry 1995; 39(1): 247-249.
7. Dabur R, Gupta A, Mandal TK, Singh DD,Bajpai V, Gurav AM, Lavekar GS.Antimicrobial activity of some indianmedicinal plants. African Journal ofTraditional, Complementary and Alternativemedicines 2005; 4(3): 313-318.
8. Uma B, Prabhakar K, Rajendran S, SarayuYL. Antimicrobial activity of Albizzialebbeckbenth against infectious diarrhoea.Internet J. Micro. 2009; 7(2009): 1.
9. Mohammed Nazneen Bobby, EdwardGnanaraj Wesely, Marimuthu AntonisamyJohnson. In vitro anti- bacterial activity ofleaves extracts of Albizia lebbeck Benthagainst some selected pathogens. AsianPacific Journal of Tropical Biomedicine.2012; S859-S862.
10. Haripriya D, Selvan N, Jeyakumar N,Periasamy R, Johnson M, Irudayaraj V. Theeffect of extracts of Selaginella in volvensand Selaginella inaequalifolia leaves onpoultry pathogens. Asian Pac. J. Trop. Med.2010; 3(9): 678-681.
11. Johnson M, Babu A, Irudayaraj V.Antibacterial studies on in vitro derived calliof Ocimum Basilicum L. J. Chem. Pharm.Res. 2011; 3(1): 715-720.
12. Paul Raj K, Irudayaraj V, Johnson M, PatricRaja D. Phytochemical and anti-bacterialactivity of epidermal glands extract ofChristella parasitica (L.) H. Lev. Asian. PacJ Trop Biomed 2011; 1(1): 8-11.
13. Lin J, Opoku AR, Geheeb- Keller M,Hutchigs AD, Terblanche SE, Jager AK, VanStaden J. Preliminary screening of sometraditional Zulu medicinal plants for anti-
Md. Nazneen Bobby et al
Current Trends in Biotechnology and PharmacyVol. 13 (4) 401-405, October 2019, ISSN 0973-8916 (Print), 2230-7303 (Online)
405
inflammatory and antimicrobial activities. J.Ethanopharmacol. 1999; 68: 267-274.
14. Ahmet Adiguzel, Medine Gulluce, MeryemSengul, Hatice Ogutcu, Fikrettin Sahin, IsaKaraman. Antimicrobial effects of Ocimumbasilicum (Labiatae) extract. Turk. J. Biol.2005; 29(1): 155-160.
15. Shahid SA and Firdous N. Antimicrobialscreening of Albizia lebbeck (l.) Benth. andAcacia leucophloea (Roxb.), African journalof pharmacy and pharmacology 2012; 6(46):3180-3183.
16. Balakumar S, Rajan S, Thirunalasundari T,Jeeva S. Antifungal activity of Aeglemarmelos (L.) Correa (Rutaceae) leaf extracton dermatophytes. Asian Pac J Trop Med.(2011); 1(4): 309-312.
17. Johnson M, Wesely EG, Zahir Hussain MI,Selvan N. In vivo and in vitro phytochemicaland anti-bacterial efficacy of Balisopermummontanum Muell. Arg. Asian Pac J TropMed. 2010; 3(11): 894-897.
18. Uzama Danlami, Envuladu Patience Elisha,A comparative study on the phytochemicalsand antimicrobial activities of the ethanol andpetroleum ether extracts of the leaves ofAlbizia Lebbeck and Its Mistletoe,International journal of pharmacy andchemistry 2017; 3(2): 13-18.
19. Ali MT, Haque ST, Kabir ML, Rana S, HaqueME, A comparative study of in vitroantimicrobial, antioxidant and cytotoxicactivity of Albizia lebbeck and Acacianilotica stem bark. Bulletin of faculty ofpharmacy, cairo university. 2018; 56: 34-38.
Screening of Antifungal Potential of Leaf Extracts from Albizia lebbeck
