|Year : 2017 | Volume
| Issue : 3 | Page : 267-272
Antifungal response of or-al-associated candidal reference strains (American Type Culture Collection) by supercritical fluid extract of nutmeg seeds for geriatric denture wearers: An in vitro screening study
Meenakshi Iyer1, Anil Kumar Gujjari2, Vishakante Gowda3, Shridhar Angadi4
1 Department of Prosthodontics, JSS Dental college and Hospital, Mysuru, Karnataka, India
2 Department of Pharmaceutics, JSS College of Pharmacy, Mysuru, Karnataka, India
3 Jagadguru Sri Shivarathreeswara University, Mysuru, Karnataka, India
4 Himalaya Drug Company, Bengaluru, Karnataka, India
|Date of Submission||11-Jan-2017|
|Date of Acceptance||19-May-2017|
|Date of Web Publication||10-Aug-2017|
Department of Pharmaceutics, JSS Dental College and Hospital, Jagadguru Sri Shivarathreeswara University, Mysuru, Karnataka
Source of Support: None, Conflict of Interest: None
Objectives: Since time immemorial, plants have continued to play a predominant role in the maintenance of human health as sources of medicinal compounds. Several effective antifungal agents are available for oral Candida infections; the failure is not uncommon because isolates of Candida albicans may exhibit resistance to the drug during therapy. The present study aimed to identify an alternative, inexpensive, simple, and effective method of preventing and controlling the candidal infection.
Methodology: All the procured and authenticated nutmeg seeds were dried in shade and cleaned by hand sorting. The crushed seeds were passed through mesh no. 40 individually. About 50 g of powdered nutmeg seeds was loaded in the supercritical fluid extractor unit using supercritical CO2 as extracting solvent in accordance with the methods of Nguyen et al. Supercritical fluid (SFE) extraction was done using CO2 gas without any cosolvents.
Results: The nutmeg extract displayed antifungal activity with the effective zone of inhibition ranging from 18.0 to 12.0 mm when compared with nystatin as positive control.
Conclusion: This paper described the in vitro antibacterial activity, and phytochemical analysis of SFE extract of nutmeg (Myristica fragrans) evaluated against C. albicans (American Type Culture Collection 10231) through agar well diffusion method. SFE of nutmeg seeds can be used as an adjunct to conventional therapy for oral candidiasis.
Keywords: Alternative medicine, antimicrobial activity, Candida albicans, complementary therapies, phytotherapy, plant extracts, supercritical fluid extraction
|How to cite this article:|
Iyer M, Gujjari AK, Gowda V, Angadi S. Antifungal response of or-al-associated candidal reference strains (American Type Culture Collection) by supercritical fluid extract of nutmeg seeds for geriatric denture wearers: An in vitro screening study. J Indian Prosthodont Soc 2017;17:267-72
|How to cite this URL:|
Iyer M, Gujjari AK, Gowda V, Angadi S. Antifungal response of or-al-associated candidal reference strains (American Type Culture Collection) by supercritical fluid extract of nutmeg seeds for geriatric denture wearers: An in vitro screening study. J Indian Prosthodont Soc [serial online] 2017 [cited 2022 May 28];17:267-72. Available from: https://www.j-ips.org/text.asp?2017/17/3/267/212730
| Introduction|| |
Fungi, once dormant and considered as nondisease-causing entity, have become active to be called as chief causative agent of various infections in health and disease. Candida albicans is accountable for most fungal infections in humans. Fungi often remain dormant and colonize various surfaces including skin and mucosa.,, Growth on animate and inanimate surfaces is a natural part of the Candida lifestyle, and it can inhabit the denture surface.
Oral candidal infections are considered opportunistic having multifactorial etiology such as drug administration (antibiotic/steroid usage), uncontrolled diabetes and/or compromised immunity, denture wearers, patients under chemotherapy, and transplant recipients.,,, According to the National Oral Health Survey, denture-related stomatitis is characterized by erythematous inflammatory areas covering the denture surface. This clinical feature is multifactorial with C. albicans being the chief etiological agent. Although rational use of synthetic drugs for treatment of candidal infections is justifiable, failure is uncommon due to candidal isolates exhibiting intrinsic (primary) or secondary resistance to the drug during therapy. With the prevalent drawbacks of antifungal drugs, their frequent use has led to drug resistance (altering drug target site, drug modification, or restricted drug penetration). The hazards and negative outcomes of medications in older people have been well documented.
To combat with antimicrobial resistance, antimicrobial alternatives can be thought as an alternative to synthetic drugs.,,, Herbal medicines have been into existence since ancient times in treating different ailments. Natural products could be an interesting alternative for the control of fungal diseases due to their lower negative impact, reduced cost, and less adverse reactions as compared to modern synthetic drugs and better patient compliance. Antibacterial, antifungal, and antioxidant activities have been observed in many plants due to the presence of secondary metabolites.
Hence, conventional preventive measures in the form of phytomedicines that have better compliance, availability, and cost need to be promoted. 60%–90% of the population has indulged in phytotherapy as a part of primary health care.
It has been cited that essential oils derived from plants have insecticidal, bactericidal, and fungicidal effects.,,,,, However, the antifungal activity of essential oils more specifically against Candida species is at infancy. This prompted us to investigate the antifungal activity of SFE extract of nutmeg seeds against Candida species.
Need of the study
- To identify essential oils from plant products which can prevent and control the growth of C. albicans
- To evaluate the antimicrobial efficacy of SFE of nutmeg seeds on C. albicans involved in causing oral candidiasis.
| Methodology|| |
Collection and identification of plant material
Nutmeg seeds were collected from a local market in Mysuru and identified by the botanist Dr. M.N. Naganadini in the Department of Pharmacognosy, JSS College of Pharmacy, Jagadguru Sri Shivarathreeswara University, SS Nagara, Mysuru, India [Table 1].
Preparation of test compound
In the present study, supercritical fluid extract (SCFE) of nutmeg seeds was prepared.
Supercritical fluid plant extract preparation
All the procured and authenticated above seeds were dried in the shade and cleaned by hand sorting. The crushed seeds were passed through mesh no. 40 individually [Figure 1]. About 50 g of powdered nutmeg seeds was loaded in the supercritical fluid extractor unit using supercritical CO2 as extracting solvent in accordance with the methods of Nguyen et al. SFE was done using CO2 gas without any cosolvents. SCFE involves the use of dense gas as a solvent (e.g., carbon dioxide [CO2]) for extraction [Figure 2]. The ground plant material was charged into the extractor. Supercritical CO2 was fed to the extractor through a high-pressure pump (300 bar) at 37°C ± 5°C, which was above its critical temperature (31°C ± 5°C) and pressure (74 bar). The extract laden CO2 was sent to a separator (60–120 bar) through a pressure reduction valve. The temperature and pressure were reduced so that the extract precipitates into the separator and gaseous CO2 is released to the atmosphere. Stock solution of the extract was prepared by dissolving 100 mg of the alcoholic extracts in 10 ml 50% of dimethyl sulfoxide (DMSO). Nystatin was used as a positive control and stock solution was prepared by dissolving 50 mg of the drug in 10 ml of DMSO to get 10 mg/ml.
All the chemicals and solvents used were procured from JSS College of Pharmacy.
Evaluation of supercritical fluid extraction of nutmeg seeds
Phytochemical analysis of SFE of nutmeg seeds were evaluated and the findings were in agreement with Trease and Evans and confirmed the presence of tannins, saponins, flavonoids, steroids, and phenols [Table 2].
|Table 2: The phytochemical screening of supercritical fluid extract of nutmeg seeds|
Click here to view
Procurement of microorganism
Freeze dried form of the microorganism C. albicans was obtained from American Type Culture Collection (ATCC 10231), Himalaya Drug Company, Makali, Bengaluru.
Preparation of inoculum
Subculture of ATCC 10231 was performed on yeast peptone dextrose at 25°C ± 2°C for 24 h to form stock culture of the microorganism (inoculum). 30 ml of molten sterile yeast peptone dextrose was poured aseptically in sterile Petri plates and was allowed to solidify at room temperature. Hundred microliters of inoculum was spread with a sterile steel spreader so as to achieve a confluent lawn of fungal growth.
Screening for antimicrobial activity
On the Petri plates with the subculture of the microorganism, wells of 6 mm were made with sterile cork borer. In one of the three plates, three aseptic wells were prepared by sterile cork borer. A 50, 100, and 150 µl volume of nutmeg extract were introduced directly into the wells (in triplicates) of the inoculated specific media agar plates. The plates were allowed to stand for 10 min for diffusion of the extract to take place and incubated at 37°C for 24 h., Sterile DMSO (50 µl, 100 µl, and 150 µl) served as the negative control in one of the Petri plates and nystatin (50 µg in 50 µl DMSO and 25 µg in 50 µl DMSO) served as the positive control in another Petri plate. The antimicrobial activity, indicated by an inhibition zone surrounding the well containing the extract, was recorded and compared with positive and negative controls. Experiments were performed in triplicates, and the mean values of the diameter of inhibition zones with ± standard deviation were calculated. The inhibition zones were measured using Hi-media zone scale available at JSS College of Pharmacy [Figure 3].
The collected data were analyzed using statistical tests such as mean value and standard deviation on an SPSS (Version 22, SPSS.inc version 16) software database.
| Results|| |
The means of the zones of inhibition of C. albicans by SFE of nutmeg seeds at 24 h were measured [Table 3].
|Table 3: Zones of inhibition in mm for Candida albicans (dimethyl sulfoxide: Nutmeg oil)|
Click here to view
Nutmeg oil with 1:1 (50 µl DMSO in 50 µl nutmeg oil) dilution with DMSO showed growth inhibition of C. albicans with 18 ± 0.13 mm followed by 1:2 concentration (50 µl DMSO in 100 µl nutmeg oil) with 12 ± 0.15 mm and with 1:3 dilution of DMSO and nutmeg oil (50 µl DMSO in 150 µl nutmeg oil) showed no zone of inhibition at 24 h, respectively; positive control nystatin at a concentration of 50 µg in 50 ml of DMSO showed zone of inhibition of 20 ± 0.14 mm and nystatin at a concentration of 25 µg in 50 µl DMSO showed zone of inhibition of 15 ± 0.12 mm [Graph 1] and [Graph 2]. In general, the plant extract showed antimicrobial activities at the critical difference of 5% (P ≤ 0.05).
| Discussion|| |
C. albicans is one among the 200 species and belongs to the genus Candida that accounts for up to 75% of all candidal infections. C. albicans is the most common Candida species residing in various regions including oral cavity and is in the benign form in health and in disease. Conventional therapeutic options for oral candidiasis range from topical polyene antifungals to azole agents. The increase in the occurrence of resistance of Candida spp. to conventional antifungals has been reported in the last few decades. Apart from this, antifungal drugs show relevant limitations such as low spectrum, interaction with other drugs, high cost, and toxic effects; the toxic effects are a result of the similarities between yeast and host cells (both eukaryotic), relevant in the clinical context. In particular, for erythematous candidiasis, the recurrence of the lesion after treatment with conventional antifungals has been reported, especially when associated with poor denture hygiene. Hence, the use of natural products can be opted as an alternative to synthetic medicine.
With various drawbacks of synthetic drugs, there is a paradigm shift from synthetic drugs to plant extracts which exhibit similar antimicrobial activity and are extensively used by the consumers due to low toxicity compared to oral care products containing antimicrobial agents., Plant extracts contain many secondary metabolites such as tannins, terpenoids, alkaloids, and flavonoids which provide new source of antimicrobial agents that could combat against drug-resistant pathogens.,,
Myristica fragrans Houtt. (family: Myristicaceae) locally known as Jaikai in Kannada is commonly in Penang Island, Malaysia, India, and Southeast Asia. Nutmeg is the dried kernel of seed of Myristica fragrans. Its pleasant aroma and warm taste make it ideal for usage as spices. It is also used as flavoring agent in beverages, syrups, and perfumes. The main constituents of M. fragrans have been found to be alkyl benzene derivatives (myristicin, elemicin, safrole, etc.), terpenes, alpha-pinene, beta-pinene, myristic acid, trimyristin,,, neolignan (myrislignan), and macelignan.
The phytochemical constituents have been investigated for antimicrobial, antidepressant, aphrodisiac, memory-enhancing, antioxidant, and hepatoprotective properties. The medicinal properties of nutmeg seeds have been extensively investigated and reported for antifungal, anticarcinogenic, anti-spasmodic, and dyspepsia.,,,, Researchers reported that trimyristin, an active compound obtained from the seed of M. fragrans, also displayed antibacterial properties against Gram-positive and Gram-negative bacteria. The antimicrobial activities of crude extracts of nutmeg seeds against oral pathogens need attention. Hence, this preliminary screening study was aimed at investigating the antimicrobial activities of SFE of nutmeg seeds against C. albicans.
Extraction of bioactive compounds from plant materials forms the main objective where a specific component is targeted which could potentiate the value of herbal medicines. Essential oil is a natural mixture extracted from several aromatic plants. A wide range of technologies could be advocated for extraction of active components from medicinal plants.
Extraction process includes conventional methods such as maceration, infusion, decoction, digestion, percolation, hot Soxhlet extraction, and distillation techniques which are based on the use of certain solvents, heat and shaking for extraction. Nonconventional methods include ultrasound, pulsed electric field, enzyme digestion, extrusion, microwave heating, and SFE. The nonconventional methods have wider advantages over conventional methods with respect to decrease in the extraction time, less usage of chemicals and solvents, and absence of residues which could cause allergic reactions and lack of thermal degradation of plant material with application of heat.,
Supercritical fluid extraction
SFE is a nonconventional method with the goal of reducing the use organic solvents and increased sample generation. SFE is the process of separating one component (the extractant) from another (the matrix) using supercritical fluids as the extracting solvent. The parameters that govern the extraction process are the temperature and pressure. The word supercritical indicates the temperature and pressure which exceed their critical values (Tc = 31.1°C, Pc = 72.8 atm) at which the substance can exist as a vapor and liquid in equilibrium. In addition, with minor changes in temperature and pressure, the substance could revert to its original form. CO2 and water are most commonly used supercritical fluids.,
SFE has gained popularity over conventional methods due to the use of a nontoxic and volatile solvent, such as CO2 which protects extracts from thermal degradation and solvent contamination.,
| Summary and Conclusion|| |
Antimicrobial resistance, relation to candidal biofilms, is a growing concern for the human population. Through overuse and misuse, antimycotic drugs have become less effective against oral pathogens. In particular, C. albicans, in the form of superficial candidal infections, have become less susceptible to standard antifungal agents making treatment much more difficult and less predictable. With the increasing size of the immunocompromised and geriatric populations, the development of potent and safe antifungal agents with as few side effects as possible is needed. It is postulated that nutmeg extract could fill this need. Even though many plants are used as home remedies for oral diseases, their antimicrobial effects on particular pathogenic microorganisms should be evaluated and implied into practice. Thus, the present screening study confirms that plant-based products interfere with the growth and metabolism of C. albicans which may prevent oral candidal infections.
Phytochemicals exhibit activities such as invulnerability for microbial growth. Hence, phytotherapy could be chosen as an alternative to allopathic medicine. The need of the hour is to promote phytomedicines and aid in the development of novel herbal preparation which is at nascent stage. Using these plant extracts as home remedies or adding to dentifrices, mouthwashes, and varnishes may create an oral environment which is unfavorable for C. albicans.
The research assessing the antimicrobial efficacy of a combination of plant extracts is the need of the hour, and such research will aid the development of a novel, innovative method that can simultaneously inhibit the two most common dental diseases of humanity, apart from lowering the development of drug resistance. Our results show high bioactivity of the SFE of nutmeg seeds against fungal pathogens and can be potential candidate for a potent antifungal molecule. The potential for developing phytomedicine into various health-care products seems to be worthwhile, both from the viewpoint of economy and safety.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Hawksworth DL. The magnitude of fungal diversity: The 1.5 million species estimate revisited. Mycol Res 2001;105:1422-32.
McMullan-Vogel CG, Jüde HD, Ollert MW, Vogel CW. Serotype distribution and secretory acid proteinase activity of Candida albicans
isolated from the oral mucosa of patients with denture stomatitis. Oral Microbiol Immunol 1999;14:183-9.
Samaranayake YH, Samaranayake LP. Experimental oral candidiasis in animal models. Clin Microbiol Rev 2001;14:398-429.
Kumamoto CA, Vinces MD. Alternative Candida albicans
lifestyles: Growth on surfaces. Annu Rev Microbiol 2005;59:113-33.
Vazquez JA, Sobel JD. Candidiasis. In: Dismukes WE, Pappas PG, editors. Clinical Mycology. New York, USA: Oxford University Press; 2003. p. 143-87.
Segal E, Baum GL. Pathogenic Yeasts and Yeast Infections. London, UK: CRC Press; 1994.
Akinyemi KO, Oladapo O, Okwara CE, Ibe CC, Fasure KA. Screening of crude extracts of six medicinal plants used in South-West Nigerian unorthodox medicine for anti-methicillin resistant Staphylococcus aureus
activity. BMC Complement Altern Med 2005;5:6.
Akpan A, Morgan R. Oral candidiasis. Postgrad Med J 2002;78:455-9.
al-Bagieh NH, Idowu A, Salako NO. Effect of aqueous extract of miswak on the in vitro
growth of Candida albicans
. Microbios 1994;80:107-13.
Al-hebshi N, Al-haroni M, Skaug N. In vitro
antimicrobial and resistance-modifying activities of aqueous crude khat extracts against oral microorganisms. Arch Oral Biol 2006;51:183-8.
Aboaba OD, Smith SI, Olude FO. Antimicrobial effect of edible plant extract on Escherichia coli
. Pak J Nutr 2006;5:325-7.
Almas K. The antimicrobial effects of extracts of Azadirachta indica
(Neem) and Salvadora persica
(Arak) chewing sticks. Indian J Dent Res 1999;10:23-6.
Almas K. The antimicrobial effects of seven different types of Asian chewing sticks. Odontostomatol Trop 2001;24:17-20.
World Health Organization. Traditional medicine growing needs and potential: WHO Policy Perspectives on Medicines, No. 002. Geneva, Switzerland: World Health Organization; 2002. p. 1-6.
Werdin González JO, Gutiérrez MM, Murray AP, Ferrero AA. Composition and biological activity of essential oils from Labiatae against Nezara viridula
) soybean pest. Pest Manag Sci 2011;67:948-55.
Patil SD, Kambli VA. Antibacterial activity of some essential oils against foodborne pathogen and food spoilage bacteria. Int J Pharm Biosci 2011;2:143-50.
Kamble VA, Patil SD. Spice-derived essential oils: Effective antifungal and possible therapeutic agents. J Herbs Spices Med Plants 2008;14:129-43.
Zabka M, Pavela R, Slezakova L. Antifungal effect of Pimenta dioica
Essential oil against dangerous pathogenic and toxigenic fungi. Ind Crops Prod 2009;30:250-3.
Leela NK, Ramana KV. Nematicidal activity of the essential oil of all spice (Pimenta dioica
L. Merr). J Plant Biol 2000;27:75-6.
Burt S. Essential oils: Their antibacterial properties and potential applications in foods – A review. Int J Food Microbiol 2004;94:223-53.
Nguyen HN, Gaspillo PD, Maridable JB, Malaluan RM, Hinode H, Salim C, et al
. Extraction of oil from Moringa oleifera kernels using supercritical carbon dioxide with ethanol for pretreatment: Optimization of the extraction process. Chem Eng Process 2011;50:1207-13.
Trease GE, Evans WC. Pharmacognosy. 13th
ed. London: Bailere Traiadal; 1989. p. 69.
Khokra SL, Prakash O, Jain S, Aneja KR, Dhingra Y. Essential oil composition and antibacterial studies of Vitex negundo
Linn. extracts. Indian J Pharm Sci 2008;70:522-6.
] [Full text]
Rios JL, Recio MC, Villar A. Screening methods for natural products with antimicrobial activity: A review of the literature. J Ethnopharmacol 1988;23:127-49.
Hammer KA, Carson CF, Riley TV. Antimicrobial activity of essential oils and other plant extracts. J Appl Microbiol 1999;86:985-90.
Aneja KR, Joshi R, Sharma C. Antimicrobial activity of Dalchini (Cinnamomum zeylanicum
bark) extracts on some dental caries pathogens. J Pharm Res 2009;2:1387-90.
Ellepola AN, Samaranayake LP. Oral candidal infections and antimycotics. Crit Rev Oral Biol Med 2000;11:172-98.
Iyer M, Gujjari AK, Rao RN, Gowda DV, Srivastava A. Biomedical applications of phytomedicines: Dental perspective. Dent Hypotheses 2016;7:34-41. [Full text]
Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12:564-82.
Ahmad I, Beg AZ. Antimicrobial and phytochemical studies on 45 Indian medicinal plants against multi-drug resistant human pathogens. J Ethnopharmacol 2001;74:113-23.
Agarwala M, Yadav RN. Phytochemical analysis of some medicinal plants. J Phytol 2011;3:10-4.
Jaiswal P, Kumar P, Singh VK, Singh DK. Biological effects of Myristica fragrans
. Ann Rev Biomed Sci 2009;11:21-9.
Qiu Q, Zhang G, Sun X, Liu X. Study on chemical constituents of the essential oil from Myristica fragrans
Houtt. by supercritical fluid extraction and steam distillation. J Chin Med Mater 2004;27:823-6.
Wang Y, Yang XW, Tao HY, Liu HX. GC-MS analysis of essential oils from seeds of Myristica fragrans
in Chinese market. China J Chin Materia Medica 2004;29:339-42.
Yang XW, Huang X, Ahmat M. New neolignan from seed of Myristica fragrans
. Zhongguo Zhong Yao Za Zhi 2008;33:397-402.
Chung JY, Choo JH, Lee MH, Hwang JK. Anticariogenic activity of macelignan isolated from Myristica fragrans
(nutmeg) against Streptococcus mutans
. Phytomedicine 2006;13:261-6.
Sonavane GS, Sarveiya VP, Kasture VS, Kasture SB. Anxiogenic activity of Myristica fragrans
seeds. Pharmacol Biochem Behav 2002;71:239-44.
Zaidi SF, Yamada K, Kadowaki M, Usmanghani K, Sugiyama T. Bactericidal activity of medicinal plants, employed for the treatment of gastrointestinal ailments, against Helicobacter pylori
. J Ethnopharmacol 2009;121:286-91.
Hussain SP, Rao AR. Chemopreventive action of mace (Myristica fragrans
, Houtt) on methylcholanthrene-induced carcinogenesis in the uterine cervix in mice. Cancer Lett 1991;56:231-4.
Ozaki Y, Soedigdo S, Wattimena YR, Suganda AG. Antiinflammatory effect of mace, aril of Myristica fragrans
Houtt. and its active principles. Jpn J Pharmacol 1989;49:155-63.
Narasimhan B, Dhake AS. Antibacterial principles from Myristica fragrans
seeds. J Med Food 2006;9:395-9.
Handa SS, Khanuja SP, Longo G, Rakesh DD. Extraction technologies for medicinal and aromatic plants. International Centre for Science and High Technology; 2008.
Sapkale GN, Patil SM, Surwase US, Bhatbhage PK. Supercritical fluid extraction. Int J Chem Sci 2010;8:729-43.
Marrone C, Poletto M, Reverchon E, Stassi A. Super critical fluids. Chem Eng Sci 1998;53:135-7.
Scalia S, Giuffreda L, Pallado P. Methods of analysis for functional foods a nutraceuticals. J Pharm Biomed Anal 1999;21:549-61.
Haslam E. Natural polyphenols (vegetable tannins) as drugs: Possible modes of action. J Nat Prod 1996;59:205-15.
[Figure 1], [Figure 2], [Figure 3]
[Table 1], [Table 2], [Table 3]
|This article has been cited by|
||In Vitro Activity of Caffeic Acid Phenethyl Ester against Different Oral Microorganisms
| ||Rasha AlSheikh, Hamad N. Albagieh, Ismail Abdouh, Hattan Zaki, Amal M. Alzahrani, Hassan S. Halawany, Khalifa S. Al-Khalifa |
| ||Applied Sciences. 2022; 12(8): 3959 |
|[Pubmed] | [DOI]|
||Evaluation of the Antimicrobial Effect of Thymoquinone against Different Dental Pathogens: An In Vitro Study
| ||Khalifa S. Al-Khalifa, Rasha AlSheikh, Moahmmed T. Al-Hariri, Hosam El-Sayyad, Maher S. Alqurashi, Saqib Ali, Amr S. Bugshan |
| ||Molecules. 2021; 26(21): 6451 |
|[Pubmed] | [DOI]|
||Chemical Composition, Antioxidant and Antimicrobial Activity of Nutmeg (Myristica fragrans Houtt.) Seed Essential Oil
| ||Vesna Nikolic, Ljubisa Nikolic, Ana Dinic, Ivana Gajic, Maja Urosevic, Ljiljana Stanojevic, Jelena Stanojevic, Bojana Danilovic |
| ||Journal of Essential Oil Bearing Plants. 2021; 24(2): 218 |
|[Pubmed] | [DOI]|
||Anti-melanogenic activity of Myristica fragrans extract against Aspergillus fumigatus using phenotypic based screening
| ||Shanu Hoda, Maansi Vermani, Rajesh K. Joshi, Jata Shankar, Pooja Vijayaraghavan |
| ||BMC Complementary Medicine and Therapies. 2020; 20(1) |
|[Pubmed] | [DOI]|