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Antibacterial and antifungal activities of Cistus incanus and C. monspeliensis leaf extracts

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Abstract

In this study, the antimicrobial activity of leaf extracts obtained from two species of genus Cistus L. was examined in vitro against five strains of bacteria and five strains of fungi. The species studied are Cistus villosus L. = incanus and Cistus monspeliensis L. All extracts showed inhibitory activity against microorganisms. These results encourage us towards further biological investigation.
... Cistus incanus aqueous solutions contain important bioactive compounds such as rutin, gallic acid, flavonoid (quercetin, kaempferol, glycoside), catechin, epicatechin, gallocatechin (Dimcheva and Karsheva, 2017). It is recorded by several studies that Cistus incanus extraxts have antibacterial and antifungal activity (Simeray et al., 1982;Chinou et al., 1994;Bouamama et al., 1999;Hannig et al., 2008;Barrajón-Catalán et al., 2010;Barros et al., 2013;Wittpahl et al., 2015;) and anti-cancer (Chinou et al., 1994;Dimas et al., 1998;Dimas et al., 2006;Hatziantoniou et al., 2006;Barrajón-Catalán et al., 2010;Skoric et al., 2012), also protective effects against DNA cleavage in cell culture (Attaguile et al., 2000). Therefore, anti-influenza virus activities in mice Ehrhardt et al., 2007;Kalusa et al., 2009) and antiviral activity against HIV and Filoviruses (Rebensburg et al., 2016) of Cistus incanus were confirmed. ...
... Apart from the high antioxidant potential phenolic extracts of Cistus originating from various species, in relation to organisms, also show a number of pro-health properties. They are i.e., anti-inflammatory, antiviral, antibacterial, and antifungal [17][18][19][20]. In recent years, were conducted an intensive research on the phenolic composition of individual cistus species and their biological activity. ...
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Long-term high fat-carbohydrates diet (HF-CD) contributes to the formation of irreversible changes in the organism that lead to the emergence of civilization diseases. In this study, the impact of three-month high-fat diet on the physical properties of erythrocytes (RBCs) was studied. Furthermore, the biological activity of Cistus Incanus L.extracts, plant known with high pro-health potential, in relation to normal and HF-CD RBCs, was determined. Obtained results have shown that, applied HF-CD modified shape, membrane potential and osmotic resistance of erythrocytes causing changes in membrane lipid composition and the distribution of lipids. The impact of HF-CD on physical properties of RBCs along with atherosclerotic lesions of the artery was visible, despite the lack of statistically significant changes in blood morphology and plasma lipid profile. This suggests that erythrocytes may be good markers of obesity-related diseases. The studies of biological activity of Cistus Incanus L. extracts have demonstrated that they may ameliorate the effect of HF-CD on erythrocytes through the membrane-modifying and antioxidant activity.
... Among the bioactive compounds to be found in the Cistus species, polyphenols, and flavonoids in particular are widely appreciated for their potential beneficial health effects (Pomponio et al., 2003;Andrade et al., 2009). These constituents exhibit a wide range of antibacterial (Chinou et al., 1994;Boumama et al., 1999), anti-inflammatory (Küpeli and Yesilada, 2007), antiviral , antiulcerogenic (Yeşilada et al., 1999), cytotoxic (Vitali et al., 2011) and antifungal activities (Chinou et al., 1994). ...
Thesis
Cistaceae family is widespread in the Mediterranean regions with several species and istraditionally known as a natural remedy. Cistus genus is present in Sardinia with populations of C.monspeliensis, C. salvifolius, C. albidus and C. creticus subspecies : C. creticus subsp. creticus, C.creticus subsp. corsicus and C. creticus subsp. eriocephalus, but few previous phytochemicalresearches have been reported on Cistus species growing in Sardinia.The aim of this research is to characterize the secondarymetabolites in extracts of differentspecies of Cistus in Sardinia and to evaluate antimicrobial and antioxydant activities. The freshaerial parts of the plants were extracted by using hydro distillation for essential oils and severaltraditional solvent s for the phenolic compounds.The chemical characterization of extracts has beenrealized by means of different chromatographic techniques such as GC/MS, HPLC DAD ESI MSand CL UHP SM/SM. Antimicrobial activity was determined as Minimum Inhibitory Concentra tionby using an agar macrodilution method. Antioxidant activity has been measured by using DPPHassay and it has been verified with EPR.A comparative analysis on the composition of essential oils showed the existence of six differentprofiles. C. cretic us subsp. eriocephalus showed a high amount of manoyl oxide and its isomer. C.salvifolius has pointed out the group of labdans; another consistent percentage is made ofperfumed molecules as ionone and its derivate. Several linear hydrocarbons were produc ed by C.monspeliensis, and the heneicosane was the most represented element. In C. albidus no labdanetype diterpenes were identified. Analysis of C. creticus subsp creticus revealed several oxygenatedsesquiterpenes and labdane type diterpenes, especiall y manoyl oxide. C. creticus subsp. corsicuswas qualitatively very similar to C. creticus subsp. creticus, notably concerning the labdane typecompounds. The analysis of the seven essential oils of Cistus creticus subsp. eriocephalus showinteresting chara cteristics and they would appear divided in two groups with different metabolicprofiles. Among solvent extracts the obtained results allowed the detection of several phenoliccompounds including phenolic acids, monomeric and dimeric flavan 3 ols, flavonolglycosides.They are characterized by a hight percentage of rosmarinic acid and derivatives and ofquercitin and derivatives. C. salvifolius is quantitatively most rich of phenolic compounds. Theextracts exhibited any pronounced differences in their anti microbial activities and revealed thatGram positive bacteria are more sensitive to the Cistus extracts than Gram negative bacteria.None of the extracts showed any noticeable action against Candida species. The extracts showedthat Cistus plants of Sardin ian origin have a greater antioxidant activity.
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Aqueous extracts of two Cistus species wild growing in Croatia—Cistus creticus (CC) and Cistus salviifolius (CS)—have been assessed with UPLC-MS/MS, showing 43 different phytochemicals, with flavonol glycosides: myricetin-3-hexoside and myricetin-rhamnoside, predominate ones in CC and myricetin-3-hexoside in CS. Antioxidant potential tested with the FRAP method showed no difference between CS and CC aqueous extracts, while higher phenolic content of CC comparing to CS, determined with a Folin–Cicolateu reagent correlated to its higher antioxidant capacity observed by the DPPH method. Both extracts were assessed for antimicrobial activity, using disc-diffusion and broth microdilution assays, targeting the opportunistic pathogens, associated with food poisoning, urinary, respiratory tract, blood stream and wound infections in humans. Antimicrobial assays revealed that fungi were in general more sensitive to both Cistus aqueous extracts, comparing to the bacteria where two extracts showed very similar activity. The most potent activity was observed against A. baumannii for both extracts. The extracts were tested on human lung cancer (A549) cell line using the MTT assay, showing very similar antiproliferative activity. After 72 h treatment with CC and CS aqueous extracts in concentration of 0.5 g/L, the viability of the cells were 37% and 50% respectively, compared to non-treated cells.
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Background Cistus munbyi Pomel is a well-known medicinal plant in Western Algeria for its several applications in medicine and food. This endemic species to North Africa is poorly studied among the genus of Cistus, which gives the opportunity to carry out original pharmacological studies to value this medicinal plant. Objective To investigate the antioxidant and the antimicrobial activities of Cistus munbyi, an endemic medicinal plant to Algeria. Methods The antioxidant properties of C. munbyi crudes extracts were evaluated using the DPPH radical-scavenging activity tests. While their antimicrobial activity was studied by the agar diffusion method and MICs determination against eighteen bacterial and fungal strains, some of which were clinical. Results The obtained results showed that aqueous and ethanol extracts possess a strong scavenging activity against DPPH radicals with EC50 values estimated at 1.12 and 8.31 µg/mL, respectively. On the other hand, extracts from C. munbyi revealed homogeneous and good antimicrobial activity, in which the inhibition zone diameters observed were ranging from 08 to 19 mm, and the MICs values were included between 0.312 and 5.000 mg/mL. Aqueous extract was the less active except against Proteus mirabilis and Staphylococcus aureus. Conclusion The findings of this study indicate that C. munbyi possess a strong antioxidant activity and an interesting antimicrobial potential, which allow justifying its uses by the local population in several applications. Therefore, C. munbyi could be as a good source of natural antioxidants and antimicrobial agents.
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Ethnopharmacological relevance: The haemorrhagic dengue fever affects up to 500 million patients, annually causing 20.000 deaths, with no chemotherapeutic agent available. The oleoresin labdanum of Cistus creticus L. has been established as an anti-infective agent since antiquity in Mediterranean ethnopharmacology. Materials and methods: We tested several extracts and fractions of labdanum - standardised on labdane-type diterpenes via GC-MS - on their activity against the dengue virus (DENV-2 strain 00st-22A) in in vitro Vero cell cultures (96-well-plates, 5 days). Preliminary experiments with a labdanum diethyl ether raw-extract did not yield measureable results due to cytotoxic effects against Vero cells. In all following experiments, cell viability was constantly checked using the MTT-test. Fractionation of this raw-extract by liquid-liquid-extraction and column-chromatography on silica-gel (gradient elution with hexane, EtOAc, CHCl3, MeOH) succeeded in separating the anti-viral activity of labdanum from its cytotoxic effect. Results: In the most active fraction GS5 at 30 μg/ml, the dengue virus proliferation was 100% suppressed and cell viability was over 90%. Structural elucidation of major constituents of GS5 is currently ongoing, but thin-layer chromatography showed that this fraction is manly dominated by manoyloxides, a class of labdane-type diterpenes with known antimicrobial activity. Claims concerning the antiviral activity of above ground parts of C. creticus have been made previously, but these generally ascribe this activity to hot water soluble polyphenols and propose an unspecific tanning effect of the viral surface proteins as the mechanism of action. However, the water soluble fraction enhanced viral proliferation. Conclusion: We therefore describe a direct, pharmacological, antiviral activity of a dichloromethane extract of labdanum against a virulent haemorrhagic fever like dengue for the first time.
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Background: Intrigued by testimonies of Saxon borreliosis self-help groups concerning considerabl improvements of their symptoms by ingestion of Cistus creticus L. (Cistaceae) leaf preparations, we recently reported on the growth inhibiting activity of extracts with different polarities and its volatile oil against Borrelia burgdorferi sensu stricto (Bbss) in vitro, determined by a bioassay guided procedure. The most active volatile oil (only about 0.10% in leaves) was found to be dominated by labdane-type manoyloxides as well as carvacrol, determined via GC-MS. Hypothesis: These terpenes are major active constituents of the old pharmaceutical oleoresin labdanum, which is secreted from the leaf surface of C. creticus and traditionally harvested, e.g., on Crete by brushing the shrubs. Methods: In order to elucidate the definite anti-Borrelia active principles of C. creticus, preparative scale separation of the diethyl‑ether soluble fraction of Cretan labdanum was achieved by combined silica gel 60-and RP-18 CC and analysed by novel TLC-Extractor/ES-MS as well as by 1d/2d-1H/13C-NMR data. For the antispirochaetal activity tests against Bbss in vitro, all samples were solubilised in water with addition of polysorbate 80, the effect of which on bacterial growth was examined and found to be negligible. Results: This led to isolation and identification of the monoterpene carvacrol as well as of the four major manoyloxides manoyloxide (A), 3-acetoxy-manoyloxide (B), 3‑hydroxy-manoyloxide (C), and epi‑manoyloxide (D). Additionally, 2-keto-manoyloxide (E) and sclareol (F) were identified via GC/EI-MS. In subsequent microbiological tests of the isolated compounds, epi‑manoyloxide (D) exhibited by far the strongest individual antispirochaetal effect, equal to the positive control amoxicilline. Furthermore, manoyloxide (A), carvacrol, and the diethyl‑ether soluble fraction of labdanum as a whole contribute to the strong antispirochaetal activity, while the other labdanes were less active. Isolated manoyloxides were further used as external standards for a GC-MS screening of labdanum samples from different origins, revealing exceptionally high contents of all analysed manoyloxides in the samples of Cretan labdanum from C. creticus, while their contents in other commercial available labdanum samples were lower by several orders of magnitude. Especially in Spanish labdanum samples, declared as Cistus ladanifer L., mainly simple alkanes and at most traces of epi‑manoyloxide (D) and of manoyloxide (A) could be detected. Conclusion: The application of C. creticus preparations by Lyme disease self-help groups may be considered as a reasonable therapy approach. For the first time, isolated epi‑manoyloxide and carvacrol could be evaluated as most promising candidates for drug development and labdanum based phytomedicine development, respectively. They should serve as vital active markers for quality assessments of C. creticus preparations.
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Some information about Cistus creticus L.
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A screening in vitro of antifungic activity of 24 strains of Basidiomycetes was realized with their culture filtrate. Lycoperdon perlatum Pers. = Pers., Oudemansiella platyphylla (pers. ex Fr.) Mos., Agrocybe dura (Bolt) Singer have shown an activity against Candida albicans, Candida tropicalis and Aspergillus fumigatus; Pholiota spumose (Fr.) Singer towards Botrytis cinerea and Lycoperdon perlatum Pers. = Pers. towards Alternaria solani, Botrytis cinerea and Verticillium dahliae. More extensive studies of the kinetics of antifungic substances production from Lycoperdon perlatum Pers. = Pers. were effectued with 2 different media and 2 different seeding technics on the strains which were sensible to the inhibitory character of this Gasteromycete.
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The antiulcer activity of a short-boiled aqueous extract from aerial parts of Cistus incanus was studied in rats against gastric lesions induced by necrotizing agents (1N HCl and absolute ethanol), indomethacin, serotonin and reserpine. The extract, containing bioflavonoids, was orally administered in the range from 0.25 to 0.50 g kg-1. It was found to have significant dose-related protective effects in all these experimental models, and was more effective against reserpine- and serotonin-induced mucosal congestion and haemorrhagic ulcers. These data suggest that the active constituents of the crude extract could be responsible for its protective effect by maintaining an efficient gastric mucosal microvascular supply.