Research Article

Determination of Antimicrobial Potential of Economically Important Persimmon (Diospyros kaki) and Date Plum (Diospyros lotus)

Sundus Malik, Dawood Ahmad*, Irfan Safdar Durrani and Muhammad Sayyar Khan

Institute of Biotechnology and Genetic Engineering (IBGE), The University of Agriculture, Peshawar, Khyber Pakhtunkhwa, Pakistan.

Received | March 26, 2024; Accepted | December 24, 2024; Published | March 11, 2025

*Correspondence | Dawood Ahmad, Institute of Biotechnology and Genetic Engineering (IBGE), The University of Agriculture, Peshawar, Khyber Pakhtunkhwa, Pakistan; Email: [email protected]

Citation | Malik, S., D. Ahmad, I.S. Durrani and M.S. Khan. 2025. Determination of antimicrobial potential of economically important persimmon (Diospyros kaki) and date plum (Diospyros lotus). Sarhad Journal of Agriculture, 41(1): 457-465.

DOI | https://dx.doi.org/10.17582/journal.sja/2025/41.1.457.465

Keywords | Diospyros kaki, Antimicrobial, Diospyros lotus, Extracts, Persimmon, Date plum

Copyright: 2025 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Traditional medicines, particularly those derived from medicinal plants, have been utilized across various cultures to address a multitude of health conditions (Partel et al., 2005). These plants contain specific bioactive compounds that may offer protection against a diverse array of microbial species. Currently, the increasing prevalence of antibiotic resistance poses a significant public health challenge and represents a major obstacle within the pharmaceutical industry. Medicinal plants are crucial in the development of new pharmaceuticals, and to fully harness their antimicrobial potential, it is essential to conduct experimental assessments of plants from various agro-climatic regions for their possible antimicrobial applications. According to Shinwari et al. (2011), approximately 12% of the more than 6,000 flowering plant species identified in Pakistan possess potential medicinal properties. Diospyros kaki (persimmon) and Diospyros lotus (date plum) are two prominent and widely cultivated members of the Ebenaceae family (Bibi et al., 2007). This family encompasses four genera and roughly 500 species. Generally, Diospyros species are deciduous trees that can reach heights of approximately 9 meters and widths of 6 meters, predominantly found in tropical and subtropical regions worldwide (Uddin et al., 2011; Singh and Joshi, 2011; Kim and Lee, 2014). The genus Diospyros is well-regarded for its multiple benefits in traditional medicine, being valued for its therapeutic properties across various cultures. Different parts of Diospyros plants are utilized for their medicinal properties; for instance, the fruits are known to alleviate flatulence and biliousness, while the leaves are employed to relieve back pain. Additionally, the bark exhibits astringent and bitter qualities, with sedative effects attributed to its seeds (Pant and Samant, 2010). Numerous studies have successfully isolated various bioactive compounds from different parts of Diospyros plants (Rice-Evans et al., 1997; Yamada et al., 1999; Prakash et al., 2000; Ahn et al., 2002; An et al., 2005; Jung et al., 2005; Chen et al., 2007; Gu et al., 2008; Igual et al., 2008; Sun et al., 2011; Takashi et al., 2011; Lee et al., 2012; Rashed et al., 2013; Ueda et al., 2013; Abozaid et al., 2014; Butt et al., 2015). The fruits of Diospyros kaki are recognized for their nutritional, laxative, stomachic, and astringent properties. The leaves possess antimicrobial, antithrombotic, and anti-wrinkle effects. The calyx and peduncle are traditionally used to treat hiccups and coughs. Conversely, the fruit of Diospyros lotus is utilized for treating constipation and acts as both a central and peripheral analgesic (Said et al., 2009). Furthermore, it exhibits antidiabetic, antitumor, antioxidant, antiseptic, laxative, sedative, nutritive, and astringent properties (Azadbakht et al., 2011). The fruit of Diospyros lotus may also be beneficial in treating dry coughs, hypertension, and diarrhoea. This study aims to conduct an antimicrobial assay on the leaf extracts from both Diospyros kaki and Diospyros lotus.

Materials and Methods

Plant material

Leaves of Diospyros kaki were collected from the Horticulture Research Farm at the University of Agriculture, Peshawar, while leaves of Diospyros lotus were sourced from the Swat district. Further analyses were conducted at the Nanobiotechnology Laboratory of the Institute of Biotechnology and Genetic Engineering.

Extraction and fractionation

The leaves from both plant species were shade-dried and subsequently ground into a homogeneous powder using an Infinigen™ Tissue Mixer Mill. A crude extract was prepared by soaking the powdered leaves in 97% methanol within a flask. This mixture was allowed to stand at room temperature for one week, with regular stirring to facilitate extraction (Chen et al., 2003). The initial methanol was filtered from the crude extract using Whatman filter paper, after which additional methanol was added to enhance the extraction of bioactive compounds. A rotary evaporator was employed to remove methanol from the crude extract at 45ºC (Sasidharan et al., 2011). The resulting crude extract for each plant was divided into two portions: one portion (10 g) was retained as the methanol crude extract, while the other portion (100 g) underwent further fractionation. For fractionation, the larger portion (100 g) of crude extracts from both Diospyros kaki and Diospyros lotus was dissolved in distilled water and transferred into a separatory funnel. The solvent-solvent partitioning was performed sequentially, starting with less polar solvents and progressing to more polar ones, specifically using n-hexane, ethyl acetate, and n-butanol to obtain n-hexane soluble, ethyl acetate soluble, and n-butanol soluble fractions, respectively. The fractions were recovered using a rotary evaporator (Rota vapor R-R 210/R215; BUCHI Labortechnik AG) and dried to yield semi-solid materials. These semi-solid extracts were further dried in a water bath at 45ºC (Kousar et al., 2023).

Culture media and microorganisms

Nutrient agar-modified media supplied by HiMedia Laboratories Pvt. Ltd. was utilized for culturing and growing microorganisms. Broth-modified media were used for incubating selected microbial strains (Supriatin et al., 2021). The bacterial strains Bacillus subtilis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, while the fungal strain Candida albicans were used in this study.

Disc diffusion method

The disc diffusion method (DDM), also known as the agar diffusion method (ADM), was employed to evaluate the sensitivity or resistance of microorganisms to antibiotics, plant extracts, and other tested samples. In this method, the plant extract diffuses over an agar medium inoculated with the test microorganism. A filter paper disc serves as a reservoir for the tested sample and is placed on the agar surface. If the plant extracts or isolated compounds exhibit antimicrobial activity, an inhibition zone will form around the disc following incubation (Horvath et al., 2016; Sharma, 2022). The diameter of this inhibition zone is indicative of the antimicrobial potency of the plant extracts or individual compounds. For testing the solvent extracts of Diospyros kaki and Diospyros lotus leaves, the disc diffusion method was applied to establish zones of inhibition in microbial cultures (Bakht et al., 2014a). Ciprofloxacin at a concentration of 50 µg/6 µl was used as a control for Gram-negative bacteria while erythromycin at 50 µg/6 µl served as a control for Gram-positive bacteria. Fluconazole at 50 µg/6 µl was used as a control for the fungal strain.

Results and Discussion

The antimicrobial potential of five different solvent extracts from the leaves of Diospyros kaki and Diospyros lotus was evaluated against four bacterial strains and one fungal strain in this study. The ethyl acetate fraction of Diospyros kaki and the methanol crude fraction of Diospyros lotus exhibited the highest zones of inhibition measuring 54% and 57%, respectively against Escherichia coli at a concentration of 3 mg disc-¹ (Figure 1). Notably, the n-hexane fraction at all tested concentrations as well as the aqueous fraction at 1 mg disc-¹ did not demonstrate any zone of inhibition (Figure 1A). The n-butanol and methanol fractions of Diospyros lotus showed significantly higher antimicrobial activity compared to the corresponding fractions of Diospyros kaki (Figure 1B). The aqueous fractions of Diospyros kaki and Diospyros lotus displayed zones of inhibition measuring 43% and 31%, respectively at a concentration of 3 mg disc-¹ (Figure 1). For Bacillus subtilis, the ethyl acetate fraction of Diospyros kaki and the n-butanol fraction of Diospyros lotus revealed the highest zones of inhibition measuring 41% and 47%, respectively at a concentration of 3 mg disc-¹ (Figure 2). The n-hexane fractions of Diospyros kaki exhibited no activity, while Diospyros lotus showed the lowest activity at all the tested fractions (Figure 2A, B). The methanol, aqueous, and n-butanol fractions from Diospyros kaki demonstrated an increasing trend in their antimicrobial activity yielding zones of inhibition measuring 34%, 35%, and 38% respectively (Figure 2A). In contrast for Diospyros lotus, aqueous, ethyl acetate and methanol fractions also exhibited a progressive increase in activity with zones measuring 34%, 35%, and 38% (Figure 2B). The ethyl acetate fraction from Diospyros kaki and n-butanol fraction from Diospyros lotus showed significant zone inhibition against Pseudomonas aeruginosa measuring 52% at a concentration of 3mg disc-¹ (Figure 3). Notably, n-butanol fraction from D. kaki exhibited significant activity at 2mg disc-¹ and 3mg disc-¹ concentrations but showed no activity concentration at 1mg disc-¹. Conversely, the methanol and n-hexane, aqueous fractions did not display any activity at any concentrations (Figure 3A).

 

 

 

 

The methanolic extract demonstrated significant activity at higher concentrations but showed no activity at the lowest concentration, similarly, both n-hexane aqueous fractions exhibited no activity across all concentrations (Figure 3B). The n-butanol fraction of Diospyros kaki leaves recorded the highest zone of inhibition against Staphylococcus aureus followed by ethyl acetate, methanolic crude, and aqueous fractions with zones measuring 43%, 40%, 34%, and 33% respectively at 3mg disc-¹ concentration. In contrast tested strain S. aureus exhibited complete resistance to n-hexane fraction across all three concentrations (Figure 4A). The ethyl acetate extract of Diospyros lotus leaves showed 38% inhibitory activity against this bacterial strain. Both n-butanol and crude samples showed zone inhibition measuring 36% at the highest concentration however methanol extract exhibited zero percent inhibition at the lowest concentration (Figure 4B). The aqueous and n-hexane fractions were comparatively less effective for inhibiting the growth of S. aureus, achieving only 25 % inhibition at 3mg disc-¹ concentration with zero percent inhibition observed at lower concentrations (Figure 4B). The antifungal activity of various extracts from leaves Diospyros kaki and Diospyros lotus against Candida albicans is illustrated in Figure 5. The crude methanol fraction of D. kaki leaves produced the highest zone of inhibition at 42%, closely followed by both ethyl acetate and n-butanol fractions, which showed similar zones of 41% at a concentration of 3 mg disc-¹. The n-hexane fraction was found ineffective against this fungal strain at any tested concentration while the aqueous fraction revealed a noticeable zone of inhibition (Figure 5A). The ethyl acetate fraction of D. lotus leaves again demonstrated significant inhibitory effects followed by methanol with an inhibition zone measuring up to 48%. Both aqueous and n-hexane fractions of D. lotus leaves displayed some level of inhibitory activity across all concentrations except the lowest concentration where no activity was noted (Figure 5B).

Diospyros kaki and Diospyros lotus are economically significant fruit crop species cultivated in various regions worldwide. This study employed a disc diffusion susceptibility assay to evaluate the antimicrobial activity of different solvent extracts of D. kaki and D. lotus leaves against four selected bacterial strains and one fungal strain used. E. coli strain exhibited a lower level of resistance in ethyl acetate, and n-butanol extracts of both D. kaki and D. lotus. These findings align with those reported by Nwaozuzu et al. (2016), and Rahman et al. (2015). At elevated concentrations (3mg disc-¹) crude extracts from D. lotus leaves demonstrated significant inhibitory efficacy consistent with previous studies. The maximum inhibitory activity (3 mg disc-¹) observed from crude extracts of D. lotus corresponds to earlier reports by Selvamohan et al. (2012). Notably a significant activity against E. coli was recorded for n-hexane-extracted leaf samples of D. lotus producing substantial zone inhibition (46%). In contrast, n-hexane fraction leaves of D. kaki showed no activity at any concentrations suggesting it lacks specific antimicrobial compounds that are effective against E. coli. These findings are supported by Ahmad et al. (2015). The highest inhibition against Bacillus subtilis growth was recorded for n-butanol extracted samples of D. lotus followed by an ethyl acetate fraction of D. kaki (3mg disc-¹). These results are consistent with earlier studies of Bakht et al. (2014b), Muthu et al. (2014), Ali et al. (2015). Crude extracts of both species exhibited significant antibacterial activity against B. subtilis at all tested concentrations compared to controls and other fractions corroborating previous findings of Chaudhary et al. (2012). Additionally, all three concentrations including aqueous extracts reduced the growth of B. subtilis indicating that except for n-hexane extracted samples, all other solvent extracts of D. kaki demonstrated inhibitory activity against this bacterium. Our results indicate that the aqueous fractions were less effective against Staphylococcus aureus however, the ethyl acetate, crude, and n-butanol

 

extracted fractions of both species exhibited inhibitory effects against this pathogen. These observations supported previous reports (Javed et al., 2015; Ejaz et al., 2014). The n-butanol fraction from D. kaki demonstrated the strongest inhibitory effect against S. aureus, followed by the ethyl acetate extract, which is consistent with the findings of Machado et al. (2003) and Abozaid et al. (2014). Crude extracts of D. lotus leaves inhibited the growth of S. aureus at all tested concentrations. Neither n-hexane extracted fractions of either species exhibited activity against Pseudomonas aeruginosa at any concentration when compared to controls and other fractions. Conversely, ethyl acetate extracted fractions of both species displayed significant inhibitory activity against P. aeruginosa resulting in zones of inhibition measuring 52% and 43%, respectively. The n-butanol fractions of both plants also demonstrated notable activity against this pathogen aligning results reported by Mamta and Monica (2016). Aqueous extracted fractions of both species showed no activity against P. aeruginosa corroborating the findings of Philip et al. (2009). Crude-extracted fractions of both species exhibited significant antifungal activities against Candida albicans, yielding zones of inhibition measuring 42% and 48%, respectively. Similar results have been reported by Toledo et al. (2015). At the highest concentration (3mg disc-¹) n-hexane fraction from D. lotus showed a zone of inhibition measuring 30% while the same fraction of D. kaki leaves did not exhibit any inhibitory effect against the fungal strain at any concentration. The ethyl acetate fractions of both species resulted in zones of inhibition measuring 41% and 50%, respectively against C. albicans. The effectiveness of ethyl acetate extracts against fungal pathogens has been previously documentedby Bakht et al. (2013). The n-butanol fractions of both species showed inhibition zones of 43% and 41% at the highest concentration inhibiting the growth of C. albicans. Similar findings were reported by Oliveira et al. (2014), and More et al. (2008). Additionally, an aqueous extracted fraction from D. kaki yielded zone inhibition measuring 41% against C. albicans while a moderate inhibition zone (28%) was recorded by an aqueous fraction from D. lotus. Overall, all derived extracts demonstrated potential antimicrobial activity, which justifies further exploration into isolating bioactive compounds.

Conclusions and Recommendations

This study has revealed that the leaves of both Diospyros kaki and Diospyros lotus have antimicrobial potential through differential activities as shown by the various extracts of these two plant species. This potential can be exploited in the future for the determination of new plant-based antimicrobial drugs.

Novelty Statement

The antimicrobial potential of economically important persimmon (Diospyros kaki) and date plum (Diospyros lotus) leaves was assessed for the first time, making this a novel research endeavor.

Author’s Contribution

Sundus Malik: Performed the experiment and wrote the article.

Dawood Ahmad and Muhammad Sayyar Khan: Designed the experiments.

Irfan Safdar Durrani: Carried out statistical analysis.

Conflict of interest

The authors have declared no conflict of interest.

References

Abozaid, H., A. Moawad, E. Amin, M. Hetta and M. Shabana. 2014. Phytochemical and biological study of Diospyros Kaki L. growing in Egypt. World J. Pharm. Res., 3(2): 1786-1795.

Ahmad, S., I. Bibi, M. Naser, A. Salam, H. Hussain, M.S. Ishaq, M. Adnan, A. Tariq and R. Ullah. 2015. Antibacterial and antifungal activities of the extract and fractions of aerial parts of Heliotropium bacciferum. Afr. J. Tradit. Complement. Altern. Med., 12(2): 32-35. https://doi.org/10.21010/ajtcam.v12i2.7

Ahn, H.S., T.I. Jeon, J.Y. Lee, S.G. Hwang, Y. Lim and D.K. Park. 2002. Antioxidative activity of persimmon and grape seed extract: In vitro and in vivo. Nutr. Res., 22(11): 1265-1273. https://doi.org/10.1016/S0271-5317(02)00429-3

Ali, N., D. Ahmad and J. Bakht. 2015. Antimicrobial activity of different solvent extracted samples from the flowers of medicinally important Plumeria obstusa. Pak. J. Pharm. Sci., 28(1): 195-200.

An, B.J., J.H. Kwak, J.M. Park, J.Y. Lee, T.S. Park, J.T. Lee, J.H. Son, C. Jo and M.W. Byun. 2005. Inhibition of enzyme activities and the antiwrinkle effect of polyphenol isolated from the persimmon leaf (Diospyros kaki folium) on human skin. Dermatol. Surg., 31(2): 848-854. https://doi.org/10.1111/j.1524-4725.2005.31730

Azadbakht, M., S.J. Hosseinimehr, M. Shokrzadeh, E. Habibi and A. Ahmadi. 2011. Diospyros lotus L. fruit extract protects G6PD-deficient erythrocytes from hemolytic injury in vitro and in vivo: Prevention of favism disorder. Eur. Rev. Med. Pharmacol. Sci., 15: 1270-1281.

Bakht, J., S. Shaheen and M. Shafi. 2014a. Antimicrobial potential of Mentha longifolia by disc diffusion method. Pak. J. Pharmacet. Sci., 27: 939-945.

Bakht, J., K. Shehla and M. Shafi. 2014b. In vitro antimicrobial activity of Allium cepa (dry bulbs) against Gram positive and Gram-negative bacteria and fungi. Pak. J. Pharma. Sci., 27: 139-145.

Bakht, J., S. Khan and M. Shafi. 2013. Anti-microbial potentials of fresh Allium cepa against gram positive and gram negative bacteria and fungi. Pak. J. Bot., 45: 1-6.

Bibi, N., A.B. Khattak and Z. Mehmood. 2007. Quality improvement and shelf life extention of persimmon fruit Diospyros kaki. J. Food. Eng., 79: 1359-1363. https://doi.org/10.1016/j.jfoodeng.2006.04.016

Butt, M.S., M.T. Sultan, M. Aziz, A. Naz, W. Ahmad, N. Kumar and M. Imran. 2015. Persimmon (Diospyros Kaki) fruit hidden phytochemicals and health claims. Exli. J., 14: 542-561.

Chaudhary, H.J., W. Shahid, A. Bano, F. Ullah, F. Munis, S. Fahad and I. Ahmad. 2012. In vitro analysis of Cupressus sempervirens L. plant extracts antibacterial activity. J. Med. Plants Res., 6(2): 273-276. https://doi.org/10.5897/JMPR11.1246

Chen, G.X.J., S.X. Xu and R.Q. Zhang. 2007. Chemical constituents of the leaves of Diospyros kaki and their cytotoxic effects. J. Asian. Nat. Prod. Res., 9(4): 347-353. https://doi.org/10.1080/10286020600727731

Chen, H.L., C.H. Wang, C.T. Chang and T.C. Wang. 2003. Effects of Taiwanese yam (Dioscorea japonica Thunb var. pseudojaponica Yamamoto) on upper gut function and lipid metabolism in Balb/c mice. Nutr., 19(7-8): 646-651. https://doi.org/10.1016/S0899-9007(03)00058-3

Dharajiya, D., P. Patel, M. Patel and N. Moitra. 2014. In vitro Antimicrobial Activity and Qualitative Phytochemical Analysis of Withania somnifera (L.) Dunal Extracts. Int. J. Pharm. Sci. Rev. Res., 27(2): 349-354.

Ejaz, R., U.A. Ashfaq and S.I. Rahat. 2014. Antimicrobial potential of Pakistani medicinal plants against multi-drug resistant Staphylococcus aureus. J. Coastal Life Med., 2(9): 714-720.

Gu, H., C. Li, Y. Xu, W. Hu, M. Chen and Q. Wan. 2008. Structural features and antioxidant activity of tannin from persimmon pulp. Food. Res. Int., 41: 208-217. https://doi.org/10.1016/j.foodres.2007.11.011

Horvath, G., T. Bencsik, K. Acs and B. Kocsis. 2016. Sensitivity of ESBL-producing gram-negative bacteria to essential oils, plant extracts, and their isolated compounds. Antibiotic Resistance. Academic Press. pp. 239-269. https://doi.org/10.1016/B978-0-12-803642-6.00012-5

Igual, M., M.L. Castello, M.D. Ortola and A. Andres. 2008. Influence of vacuum impregnation on respiration rate mechanical and optical properties of cut persimmon. J. Food. Eng., 86: 315-323. https://doi.org/10.1016/j.jfoodeng.2007.06.002

Javid, T., M. Adnan, A. Tariq, B. Akhtar, R. Ullah and N.M. Elsalam. 2015. Antimicrobial activity of three medicinal plants (artemisia indica, medicago falcata and tecoma stans). Afr. J. Tradit. Complement. Altern. Med., 12(3): 91-96. https://doi.org/10.4314/ajtcam.v12i3.11

Jung, S.T., Y.S. Park, Z. Zachwieja, M. Folta, H. Barton and J. Piotrowicz. 2005. Some essential phytochemicals and the antioxidant potential in fresh and dried persimmon. Int. J. Food. Sci. Nutr., 56: 105-113. https://doi.org/10.1080/09637480500081571

Kim, M.G. and H.S. Lee. 2014. 1, 2-Benzendiol isolated from persimmon roots and its structural analogues show antimicrobial activities against food born bacteria. J. Korean Soc. Appl. Biol. Chem., 57(4): 429-433. https://doi.org/10.1007/s13765-014-4141-x

Kousar, Z., S. Manzoor, S. Naz and M.S. Shahid. 2023. Extraction, fractionation, phytochemical profile, antioxidant, and antimicrobial activities of Cassia absus L. and Citrus medica L. Pharma. Biomed. Res., 9(2): 133-146. https://doi.org/10.32598/PBR.9.2.1121.1

Lee, J.H., Y.B. Lee, W.D. Seo, S.T. Kang, J.W. Lim and K.M. Cho. 2012. Comparative studies of antioxidant activities and nutritional constituents of persimmon juice (Diospyroskaki L. cv. Gapjubaekmok). Prev. Nutr. Food Sci., 17: 141-151. https://doi.org/10.3746/pnf.2012.17.2.141

Machado, T.B., A.V. Pinto, M.C. Pinto, I.C.R. Leal, M.G. Salvia and A.C.F. Amaral. 2003. In vitro activity of Brazilian medicinal plants, naturally occurring naphthoquinones and their analogues against methiclin-resistant staphylococcus aureus. Int. J. Antimicrb. Agent, 21(3): 279-284. https://doi.org/10.1016/S0924-8579(02)00349-7

Mamta, G. and S. Monica. 2016. Assay of antimicrobial activity of roots extracts of Oroxylum indicum (L.) vent. against seven pathogenic bacteria. Int. J. Boil. Pharma. Res., 7(3): 166-171.

More, G.K., T.E. Tshikalange, N. Lall, F.S. Botha, J.M. Meyer. 2008. Antimicrobial activity of medicinal plants against oral microorganisms. J. Ethnol. Pharmacol., 119(1): 473-477. https://doi.org/10.1016/j.jep.2008.07.001

Muthu, K., M. Hema, S. Nagaraj and R. Rengasamy. 2014. In vitro antibacterial potential, phytochemical characterization of Cyperus rotundus flower extract. Int. J. Natl. Prod. Res., 4(1): 6-8.

Nwaozuzu, E.E. and G.C. Ebi. 2016. Antimicrobial and therapeutic potentials of the ethyl acetate fractions of crude methanolic extract of Monodora myristica seed. Br. J. Pharma. Res., 11(5): 1-8. https://doi.org/10.9734/BJPR/2016/23227

Oliveira, G.T.D., J.M.S. Ferreira, L.H. Rosa, E.P.D. Siqueira, S. Johann and L.A. Lima. 2014. In vitro antifungal activities of leaf extracts of Lippia alba (Verbenaceae) against clinically important yeast species. Rev. Soc. Bras. Med. Trop., 47(2): 247-250. https://doi.org/10.1590/0037-8682-0008-2013

Pant, S. and S.S. Samant. 2010. Ethnobotanical observation in the Momaula reserve forest of Koumoun West Himalaya, India. Ethnobotanical Leaflets. 2010 (2). Available at: https://opensiuc.lib.siu.edu/ebl/vol2010/iss2/8

Partel, M., R. Kalamees, U. Reier, E. Tuvi, E. Roosaluste, A. Vellak and M. Zobel. 2005. Grouping and prioritization of vascular plant species for conservation: Combining natural rarity and management need. Biol. Cons., 123: 271-278. https://doi.org/10.1016/j.biocon.2004.11.014

Philip, K., S.N.A. Malek, W. Sani, S.K. Shin, S. Kumar, H.S. Lai, L.G. Serm and S.N.S.A. Rahman. 2009. Antimicrobial Activity of Some Medicinal Plants from Malaysia. Am. J. Appl. Sci., 6(8): 1613-1617. https://doi.org/10.3844/ajassp.2009.1613.1617

Prakash, P., N.I. Krinsky and R.M. Russell. 2000. Retinoids, carotenoids and human breast cancer cell cultures: A review of differential effects. Nutr. Rev., 58: 170-176. https://doi.org/10.1111/j.1753-4887.2000.tb01856.x

Rashed, K.N., R.C.F. Cheung and T.B. Ng. 2013. Bio-active phytoconstituents from non-polar extracts of Diospyros lotus stems and demonstration of antifungal activity in the extracts. World. J. Pharm. Sci., 1(4): 99-108.

Rehman, A., A. Rehman and I. Ahmad. 2015. Antibacterial, antifungal, and insecticidal potentials of Oxalis corniculata and its isolated compounds. Int. J. Anal. Chem., 2015: 1-5. https://doi.org/10.1155/2015/842468

Rice-Evans, C.A., N.J. Miller and J. Paganga. 1997. Antioxidant properties of phenolic compounds. Trends Plant. Sci., 2: 152-159. https://doi.org/10.1016/S1360-1385(97)01018-2

Said, A., U.W. Hawas, S.M. Nofal, K. Rashed and A. Huefner. 2009. Pharmaco-chemical studies on the aqueous methanolic extract of Diospyros lotus leaves. Res. J. Phytochem., 3(1): 1–12. https://doi.org/10.3923/rjphyto.2009.1.12

Sasidharan, S., Y. Chen, D. Saravanan, K.M. Sundram and L.Y. Latha. 2011. Extraction, isolation, and characterization of bioactive compounds from plants extracts. Afr. J. Trad. Comp. Altern. Med., 8(1): 1-10. https://doi.org/10.4314/ajtcam.v8i1.60483

Selvamohan, T., V. Ramadas, S. Shibila and S. Kishore. 2012. Antimicrobial activity of selected medicinal plants against some selected human pathogenic bacteria. Adv. Appl. Sci. Res., 3(5): 3374-3381.

Sharma, R., 2022. Kirby Bauer disc diffusion method for antibiotic susceptibility testing. Microbe Notes, https://microbenotes.com/kirby-bauer-disc-diffusion/

Shinwari, S., R. Qureshi and E. Baydoun. 2011. Ethnobotanical study of Kohat Pass (Pakistan). Pak. J. Bot., 43: 135-139.

Singh, S. and H. Joshi. 2011. Diospyros kaki (Ebenaceae): A review. Asian. J. Res. Pharma. Sci., 1(3): 55-58.

Sun, L., J. Zhang, X. Lu, L. Zhang and Y. Zhang. 2011. Evaluation to the antioxidant activity of total flavonoids extract from persimmon (Diospyros kaki L.) leaves. Food Chem. Toxicol., 49: 2689-2696. https://doi.org/10.1016/j.fct.2011.07.042

Supriatin, Y., V.A. Sumirat and M. Herdiani. 2021. Growth analysis of Escherichia coli and Salmonella typhi on MacConkey agar modification. J. Phys. Conf. Ser., 1764: 012207. https://doi.org/10.1088/1742-6596/1764/1/012207

Takashi, A., T. Tomoyuki, I. Ayako and Y. Keizo. 2011. Effects of seasonal temperature changes on DkMyb4 expression involved in pro anthocyanidin regulation in two genotypes of persimmon (Diospyros kaki Thunb.) fruit. Planta, 233(5): 883-894. https://doi.org/10.1007/s00425-010-1346-z

Toledo, C.E.M.D., P.R. Santos, J.C.P.D. Mello, B.P.D. Filho, C.V. Nakamura and T. Ueda-Nakamura 2015. Antifungal properties of crude extracts, fractions, and purified compounds from bark of Curatella Americana L. (Dilleniaceae) against Candida Species. Evid. Based Complement. Altern. Med., 2015: 1-9. https://doi.org/10.1155/2015/673962

Uddin, G., A. Rauf, B.S. Siddiqui, M. Arfan and I.U. Rahman. 2013. Proximate chemical composition and antimicrobial activities of fixed oils from Diospyros lotus L. Med. Chem., 3: 282-285. 

Uddin, G., T.U. Rehman, M. Arfan, W. Liaqat and Waliullah. 2011. Antimicrobial, insecticidal and phytotoxic activities of Indigofera heterantha roots. J. Med. Plants Res., 5: 5835-5839.

Ueda, K., R. Kawabata, T. Irie, Y. Nakai, Y. Tohya and T. Sakaguchi. 2013. Inactivation of viruses by plant-derived tannins: strong effects of extracts from persimmon (Diospyros kaki) on a broad range of viruses. PLoS One J., 8(1): 1371-1381. https://doi.org/10.1371/journal.pone.0055343

Yamada, Y., A. Yamamoto, N. Yoneda and N. Nakatani. 1999. Identification of kaempferol from the leaves of Diospyros kaki and its antimicrobial activity against Streptococcus mutans. Biocontr. Sci., 4(2): 97-100. https://doi.org/10.4265/bio.4.97