Investigation of Antifungal and Antioxidant Properties of Capparis ovata Methanolic Extracts

Caper plant belonging to the genus Capparis (family Capparaceae) is a plant used in traditional medicine to cure various illnesses since ancient times. Studies have shown significant medicinal properties of various Capparis species. This study was designed to examine in vitro antifungal and antioxidant activity of methanolic extracts of Capparis ovata buds. It was determined that C. ovata inhibited the NO radical in a dose-dependent manner and exhibited reducing power activity. According to the standard pyrocatechol graph, 1 mg of C. ovata contains 19.64 μg of phenolic equivalent of pyrocatechol. In vitro antifungal susceptibility testing of C. ovata methanolic extract against eight different fungal strains was determined by broth macrodilution method. According to MIC values of in vitro antifungal activity testing C. ovata has moderate antifungal activity. In conclusion, C. ovata was found to have a significant antifungal and antioxidant potential. Hence, this plant could have the potential to be used against fungal and oxidative stress related many disease conditions.

properties (Duman and Ozcan, 2014;Belviranlı et al., 2016).This plant belongs to the family of Capparaceae and to the genus Capparis with 250 and 400 species.They are grown generally in tropical and subtropical regions but some also are in temperate regions in the Mediterranean including Turkey (Lansky et al., 2013).Capparis ovata have wide natural distribution, and it is cultivated in large parts of Turkey (Arslan et al., 2010;Hacıseferogulları et al., 2011).Different parts of capper plant have been used since ancient times for cosmetic, nutritional and medicinal purposes.In traditional medicine Capparis species were used to treat some disorders including rheumatic diseases, stomach problems, headache and toothache (Lansky et al., 2013;Tlili et al., 2011).Caper plant has different pharmacological activities and its various parts are used for pharmaceutical purposes (Arslan et al., 2010;Kondawar et al., 2011).The Capparaceae family generally contains glucosinolates, alkaloids and flavonoids, but the phytochemical composition of the contents obtained from different plant parts varies (Arslan et al., 2010;Tlili et al., 2011).A number of studies have shown that capparis species have significant immunostimulant, antitumoral, antidiabetic, antisclerosis, antimicrobial, antioxidative, antiinflammatory, immunomodulatory and antiviral activities.However, additional studies are needed to validate the use of Capparis species in medical treatment (Tlili et al., 2011;Zia-Ul-Hag et al., 2011;Mishra et al., 2007).Studies indicate that the incidence and prevalence of some mycoses remains to be a public health challenge.Increased use of antifungal drugs has led to the resistance problems in today's world.In addition, due to increased number of multidrugresistant fungal strains and limited number of antifungal agents, it is necessary to discover new antifungals from natural products (Aqil et al., 2010).Some medicinal plants possess potentially great antioxidant activity.Antioxidants are capable of preventing or protecting from the oxidative stress in cells.Hence, they have beneficial effects in the treatment of many diseases.Medicinal plants are a good source of natural antioxidants, which are alternative ways to use of synthetic antioxidants.They are also very good candidate for side effect free alternatives in comparison to synthetic antioxidants to be used in food industry and in preventive medicine (Ali et al., 2008;Krishnaiah et al., 2011).Therefore, it is aimed to investigate the antioxidant and antifungal activities of C. ovata species in this study.

Preparation of Plant Material and Extracts
The buds of C. ovata were obtained from a local herbal shop in Kayseri Province (Gül Gıda ve Tarım Ürünleri) Turkey.Methanolic extracts of the buds were prepared according to the method described by Özkan et al. (2013) were stored at -25 °C until used in the experiments.

Test Microorganisms:
The antifungal activities of C. ovata methanolic extracts were evaluated against different types of standard fungal strains including yeasts and filamtous fungi.Candida albicans (ATCC 90028), Candida parapsilosis (ATCC22019), Candida krusei (ATCC 6258), Malassezia pachydermatis yeast strains and Microsporum canis, Microsporum gypseum, Microsporum nanum and Trichophyton mentagrophytes filamentous fungi strains were used as test microorganisms and provided from the culture collection of Istanbul University Faculty of Veterinary Medicine Department of Microbiology.

In Vitro Antifungal Susceptibility Testing:
Reference methods were used according to the guidelines of the Clinical and Laboratory
ISSN: 2148-2837/ MAKU J. Health Sci.Inst.susceptibility testing.Broth macrodilution assay was performed for determination of minimum inhibitory concentration (MIC) values of the methanolic extract of C. ovata buds using the guidelines CLSI M27-A3 for yeasts and CLSI M38-A2 for filamentous fungi (CLSI, 2008a;CLSI, 2008b).Broth macrodilution assay was also performed for Amphotericin B as a reference antifungal agent.

NO Radical Scavenging Activity
Nitric oxide (NO) radical scavenging activity of C. ovata extract was measured according to the methods of Badami et al. (2003) and Kumar et al. (2005) with slight modification.The reaction is based on production of NO by sodium nitroprusside at physiological pH.Nitric oxide interacts with oxygen to generate nitrite.Nitrite ions form a colored product by Greiss reaction, which is read at 548 nm to determine the NO level (Green et al., 1982).

Reducing Power Capacity
The reducing power of C. ovata extract was measured by the method of Oyaizu (1986).

Determination of Total Phenolic Compounds
Total soluble phenolic content of C. ovata extract was measured using Folin-Ciocalteu reagent according to the method of Slinkard and Singleton (1977).

Results
The C. ovata methanolic extract at different amounts was compared with Rutin standard for NO radical scavenging activity (Figure 1). Figure 2  Reducing power

Discussion
Previous studies indicate that the antioxidant activity of the C. ovata and other capparis species have high anti-oxidant activity (El-Ghorab et al., 2007;Naziroğlu et al., 2011;Unver et al., 2009;Duman et al., 2013;Naziroğlu et al., 2013).In this study, the results of radical scavenging activity of the C. ovata extract and the reducing power and phenolic compound content indicates that this plant is a potential antioxidant.Our results are similar to previous works studying other Capparis species.Zia-Ul- Haq et al. (2011) reported that C. decidua extracts have prominently high levels of phenolic compounds and showed potent antioxidant activity and reduced different types of radicals.El-Ghorab et al. (2007) showed that the methanolic extract of Capparis spinosa has a strong antioxidant activity.The ethyl acetate and aqueous extract of C. spinosa was reported to have DPPH scavenging activities (Yang et al., 2010).Due to these properties, C. ovata has the potential to be used as a natural antioxidant source in both pharmaceutical and food industry.Several studies indicate that many Capparis species have the antifungal activities against several fungal pathogens (Naziroğlu et al., 2011;Aslam et al. 2010;Sharma et al., 2009;Keymanesh et al., 2009;Malabadi et al., 2007;Buwa et al., 2006;Anywar et al., 2014;Rathee et al., 2013).However, there is lack of studies about antifungal activity of the species C. ovata.There are the reports that other Capparis species have antifungal activity.Capparis spinosa was assayed for antifungal activity toward the phytopathogenic fungi Valsa vali by Lam and Tzi-Bun Ng (2009).According to these results of research C. spinosa inhibited the reproduction of this fungus.Aslam et al. (2010) have tested the Capparis decidua against different fungus strains.In their study, they found that this herb is effective against to Rhizoctonia solani.According to in vitro antifungal susceptibility testing results of this study, C. ovata has moderate antifungal activity.In conclusion, this study indicates that C. ovata has an antioxidant and antifungal effect and is worth considering in therapeutic formulations to cure several diseases.However further studies are recommended to verify the role of this plant in herbal medicine for discovering new natural bioactive pharmaceuticals.

Figure 1 .
Figure 1.Inhibition of nitric oxide (NO) radicals by Capparis ovata and Rutin standard.

Table 1 :
In vitro antifungal susceptibility of Capparis ovata methanolic extract and amphotericin B against fungal test strains.
illustrates the reducing power capacity of C. ovata extract in comparison to butylated hydroxyl toluene (BHT).The reducing power of C. ovata methanol extract increased with the increasing amounts of sample.