Heavy Metals Content in Selected Medicinal Plants Produced and Consumed in Serbia and Their Daily Intake in Herbal Infusions.

The heavy metals content (HMs) was investigated in 14 different medicinal plants collected from the three regions in Central Serbia, Zlatar, Sokobanja, and Kopaonik. The concentrations of Cd, Cr, Ni, Hg and Pb were determined: Cd (<0.03-2.72 mg/kg); Cr (<0.08-12.1 mg/kg); Ni (<0.08-12.2 mg/kg); Pb (0.6-49.0 mg/kg); the Hg concentration was below the detection limit of 0.06 mg/kg in all samples. The daily intake of HMs due to ingestion of 200 mL of herbal infusion was in all cases below the recommended limit prescribed by the World Health Organization. The estimated daily intake values were below the values for the oral reference dose regulated by the U.S. Environmental Protection Agency (USA EPA). The target hazard quotient and hazard index for Cd, Cr Ni, and Pb were below 1. Nevertheless, due to the tendency of heavy metals to accumulate in the organism, attention should be paid to the daily intake of herbal infusion during long-term usage. Specifically, it is recommended to consume not more than one cup (200 mL) of infusion per day made from thyme (Mt. Zlatar) and blueberry (Mt. Kopaonik), and not more than two cups per day for other herbs.

An Insight into an Olive Scab on the "Istrska Belica" Variety: Host-Pathogen Interactions and Phyllosphere Mycobiome.

The olive tree is one of the most important agricultural plants, affected by several pests and diseases that cause a severe decline in health status leading to crop losses. Olive leaf spot disease caused by the fungus Venturia oleaginea can result in complete tree defoliation and consequently lower yield. The aim of the study was to obtain new knowledge related to plant-pathogen interaction, reveal mechanisms of plant defense against the pathogen, and characterize fungal phyllosphere communities on infected and symptomless leaves that could contribute to the development of new plant breeding strategies and identification of novel biocontrol agents. The highly susceptible olive variety "Istrska Belica"' was selected for a detailed evaluation. Microscopy analyses led to the observation of raphides in the mesophyll and parenchyma cells of infected leaves and gave new insight into the complex V. oleaginea pathogenesis. Culturable and total phyllosphere mycobiota, obtained via metabarcoding approach, highlighted Didymella, Aureobasidium, Cladosporium, and Alternaria species as overlapping between infected and symptomless leaves. Only Venturia and Erythrobasidium in infected and Cladosporium in symptomless samples with higher abundance showed statistically significant differences. Based on the ecological role of identified taxa, it can be suggested that Cladosporium species might have potential antagonistic effects on V. oleaginea.

New perspectives of purple starthistle (Centaurea calcitrapa) leaf extracts: phytochemical analysis, cytotoxicity and antimicrobial activity.

Ethnobotanical and ethnopharmacological studies of many Centaurea species indicated their potential in folk medicine so far. However, investigations of different Centaurea calcitrapa L. extracts in terms of cytotoxicity and antimicrobial activity against phytopathogens are generally scarce. The phenolic profile and broad antimicrobial activity (especially towards bacterial phytopathogens) of methanol (MeOH), 70% ethanol (EtOH), ethyl-acetate (EtOAc), 50% acetone (Me2CO) and dichloromethane: methanol (DCM: MeOH, 1: 1) extracts of C. calcitrapa leaves and their potential toxicity on MRC-5 cell line were investigated for the first time. A total of 55 phenolic compounds were identified: 30 phenolic acids and their derivatives, 25 flavonoid glycosides and aglycones. This is also the first report of the presence of centaureidin, jaceidin, kaempferide, nepetin, flavonoid glycosides, phenolic acids and their esters in C. calcitrapa extracts. The best results were obtained with EtOAc extract with lowest MIC values expressed in µg/mL ranging from 13 to 25, while methicillin resistant Staphylococcus aureus was the most susceptible strain. The most susceptible phytopathogens were Pseudomonas syringae pv. syringae, Xanthomonas campestris pv. campestris and Agrobacterium tumefaciens. The highest cytotoxicity was recorded for EtOAc and Me2CO extracts with the lowest relative and absolute IC50 values between 88 and 102 µg/mL, while EtOH extract was the least toxic with predicted relative IC50 value of 1578 µg/mL. Our results indicate that all tested extracts at concentration considered as non-toxic can be one of great importance in combat towards phytopathogenic and human pathogenic strains, as well as natural sources of antimicrobials.

Balkans' Asteraceae Species as a Source of Biologically Active Compounds for the Pharmaceutical and Food Industry.

Herbal drugs are a useful source of different bioactive compounds. Asteraceae species, as the most widespread vascular plants, can be used both as food and as medicine due to the great diversity of recorded chemical components - different phenolic compounds, terpenes, carotenoids, vitamins, alkaloids, etc. The Balkan Peninsula is characterized by great diversity of plants from Asteraceae family, including presence of rare and endemic species. In this review, results of the survey of chemical composition and biological activity, mainly focusing on antioxidant, antimicrobial and anticancer effects of selected Balkans' Asteraceae species were provided. In addition, information on edible plants from Asteraceae family is presented, due to growing interest for the so-called 'healthy diet' and possible application of Balkans' Asteraceae species as food of high nutritional value or as a source of functional food ingredients.

Activity concentrations of 137Cs, 40K, and 210Pb radionuclides in selected medicinal herbs from Central Serbia and their effective dose due to ingestion.

Specific activity of 137Cs, 40K, and 210Pb radionuclides in fifteen selected medicinal herbs from three locations in Central Serbia (two mountains, Kopaonik and Zlatar, and a valley, Sokobanja) was measured using two semiconductor HPGe spectrometer systems. The obtained values are in intervals (<0.3 ÷ 9.7) Bq/kg, (<0.2 ÷ 24.7) Bq/kg, and (<0.2 ÷ 5.7) Bq/kg for 137Cs; in intervals (125 ÷ 1100) Bq/kg, (104 ÷ 872) Bq/kg, and (103 ÷ 954) Bq/kg for 40K, and in intervals (3.6 ÷ 49.0) Bq/kg, (3.9 ÷ 57.9) Bq/kg, and (2.8 ÷ 103) Bq/kg for 210Pb, for herbs from Kopaonik, Sokobanja and Zlatar, respectively. The highest activity measured in individual herbs was: 24.7 Bq/kg for 137Cs (Sokobanja valley), 1100 Bq/kg for 40K (Mt. Kopaonik) and 103 Bq/kg for 210Pb (Mt. Zlatar). The corresponding individual annual effective doses due to ingestion calculated from the measured activity concentrations, of the radionuclides are: in intervals (<1.7 ÷ 82.9) nSv, (<1.4 ÷ 211) nSv, and (<1.1 ÷ 48.7) nSv for 137Cs; in intervals (0.76 ÷ 4.5) µSv, (0.64 ÷ 4.4) µSv, and (0.63 ÷ 4.9) µSv for 40K, and in intervals (1.1 ÷ 18.2) µSv, (1.3 ÷ 21.6) µSv, (0.9 ÷ 38.3) µSv for 210Pb, respectively. The specific activity concentration values of 137Cs and 40K obtained for the investigated herbs are similar to the literature data, while no information was found in literature about specific activity concentration of 210Pb. The all values obtained for individual annual effective dose due to ingestion are less than 100 µSv, which means that the daily use of 200 mL of herbal infusion during a year made from the investigated herbs does not represent a radiological risk for health. However, a high individual annual effective dose of ingestion of 210Pb obtained for some herbs indicate that their consumption in larger amounts 2-3 cups of infusion daily over a year could make the dose to exceed the recommended level, and points to necessity for extensive investigation of 210Pb activity concentration worldwide.

Frankincense and myrrh essential oils and burn incense fume against micro-inhabitants of sacral ambients. Wisdom of the ancients?

ETHNOPHARMACOLOGICAL RELEVANCE: Essential oils obtained from resins of Boswellia carteri Birdw. and Commiphora myrrha (Nees) Engl., commonly known as frankincense and true myrrh respectively, have been used extensively since 2800 BCE for the treatment of skin sores, wounds, teeth, inflammation, and urinary tract diseases in traditional medicine; for preparation of mummification balms and unguents; and also as incense and perfumes. Since ancient times, burning of frankincense and myrrh in places of worship for spiritual purposes and contemplation (a ubiquitous practice across various religions) had hygienic functions, to refine the smell and reduce contagion by purifying the indoor air. AIM OF THE STUDY: The general purpose of the study was to assess the in vitro antimicrobial potential of the liquid and vapour phases of B. carteri and C. myrrha essential oils and burn incense, as well as to test the effectiveness of their in situ application to cleanse microbially-contaminated air within the ambient of an investigated 17th-century church. MATERIALS AND METHODS: The chemical composition of B. carteri and C. myrrha essential oils, obtained by hydrodistillation of frankincense and true myrrh oleo gum resins was determined using GC/MS, and antimicrobial properties of their liquid and vapour phases were assessed by the broth microdilution and microatmosphere diffusion methods. Chemical analysis of burn incense fume obtained using bottle gas washing with dichloromethane as a solvent was performed by GC/MS, while its antimicrobial activity was evaluated using a modified microatmosphere diffusion method to evaluate germination inhibition for fungi and CFU count reduction for bacteria. The in situ antimicrobial activity of B. carteri burn incense and essential oil vapour phase was assessed in the sealed nave and diaconicon of the church, respectively. RESULTS: The dominant compounds of B. carteri EO were α-pinene (38.41%) and myrcene (15.21%), while C. myrrha EO was characterized by high content of furanoeudesma-1,3-diene (17.65%), followed by curzerene (12.97%), ß-elemene (12.70%), and germacrene B (12.15%). Burn incense fume and soot had α-pinene (68.6%) and incensole (28.6%) as the most dominant compounds, respectively. In vitro antimicrobial assays demonstrated high bacterial and fungal sensitivity to the liquid and vapour phases of EOs, and burn incense fume. In situ application of B. carteri EO vapour and incense fume resulted in reduction of air-borne viable microbial counts by up to 45.39 ±â€¯2.83% for fungi and 67.56 ±â€¯3.12% for bacteria (EO); and by up to 80.43 ±â€¯2.07% for fungi and 91.43 ±â€¯1.26% for bacteria (incense fume). CONCLUSIONS: The antimicrobial properties of essential oil derived from frankincense, a compound with well-known traditional use, showed that it possesses a clear potential as a natural antimicrobial agent. Moreover, the results suggest possible application of B. carteri EO vapour and incense fume as occasional air purifiers in sacral ambients, apart from daily church rituals.

Pollution by Urticaceae pollen-influence of selected air pollutants and meteorological parameters.

The goal of this study was to analyze the influence of pollutants (concentrations of NO2, SO2, and soot in the air) and meteorological parameters (air temperature, humidity, wind speed, air pressure, cloud index) on Urticaceae pollen type emission measured in the region of Subotica, Serbia. The concentrations of the air pollutants, Urticaceae pollen, and meteorological parameters were measured over a 5-year period (2009-2013), followed by a statistical analysis of the values obtained. For most of the years examined, the concentration of NO2 correlates significantly with the concentration of Urticaceae pollen type. It was also established that air temperature, humidity, wind speed, atmospheric pressure, and cloud index have an influence on Urticaceae pollen type emission, while SO2 and soot do not contribute.

Essential-oil variability of Juniperus deltoides R.P.Adams along the east Adriatic coast - how many chemotypes are there?

The composition of the essential oils isolated from twigs of ten Juniperus deltoides R.P. Adams populations from the east Adriatic coast was determined by GC-FID and GC/MS analyses. Altogether, 169 compounds were identified, representing 95.6-98.4% of the total oil composition. The oils were dominated by monoterpenes (average content of 61.6%), which are characteristic oil components of species of the Juniperus section. Two monoterpenes, α-pinene and limonene, were the dominant constituents, comprising on average 46.78% of the essential oils. Statistical methods were deployed to determine the diversity of the terpene classes and the common terpenes between the investigated populations. These statistical analyses revealed the existence of three chemotypes within all populations, i.e., a α-pinene, limonene, and limonene/α-pinene type.

Variability of the needle essential oils of Juniperus deltoides R.P.ADAMS from different populations in Serbia and Croatia.

The essential-oil compositions of one Croatian and three Serbian populations of Juniperus deltoides R.P.ADAMS have been determined by GC/MS analysis. In total, 147 compounds were identified, representing 97.3-98.3% of the oil composition. The oils were dominated by monoterpenes, which are characteristic components for the species of the section Juniperus. Two monoterpenes, α-pinene and limonene, were the dominant constituents, with a summed-up average content of 49.45%. Statistical methods were used to determine the diversity of the terpene classes and the common terpenes between the newly described J. deltoides populations from Serbia and Croatia. Only reports on several specimens from this species have been reported so far, and there are no studies that treat population diversity. Cluster analysis of the oil contents of 21 terpenes showed high correlation with the geographical distribution of the populations, separating the Croatian from the Serbian populations. The comparison of the essential-oil compositions obtained in the present study with literature data, showed the separation of J. deltoides and J. oxycedrus ssp. oxycedrus from the western Mediterranean region.

Composition and antimicrobial activity of Seseli globiferum essential oil.

The essential oil from aerial parts of Seseli globiferum Vis. obtained by hydrodistillation with Clevenger-type apparatus was analyzed by GC-MS. Twenty-eight compounds were identified, representing 99.4% of the total oil. The main components of the oil were sabinene (38.0%), alpha-pinene (21.2%) and beta-phellandrene (13.5%). The microbial growth inhibitory properties of the isolated essential oil were determined using the broth microdilution method against seven bacterial species: Salmonella typhimurium (ATCC 13311), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Enterobacter cloacae (clinical isolates), Bacillus cereus (clinical isolates), Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 12228), Micrococcus flavus (ATCC 10240) and three fungal species: Aspergillus niger (ATCC 6275), Aspergillus versicolor (ATCC 11730), Trichoderma viride (IAM 5061) and Penicillium funiculosum (ATCC 36839). The essential oil showed activity against bacteria P. aeruginosa, followed by M flavus, L. monocytigenes and E. coli, and all investigated fungal species.

Composition and antifungal activity of the essential oil of Seseli annuum wild-growing in Serbia.

The essential oil from the aerial parts of Seseli annuum, wild-growing in Serbia, was obtained by hydrodistillation and analyzed using GC and GC/MS. A total of 43 components were identified representing 96.5% of S. annuum oil. The most abundant compounds were germacrene D (29.8%), sabinene (10.3%), beta-ocimene Z (9.8%) and limonene (8.6%). The essential oil showed antifungal activity against fifteen fungi with MICs between 12.5 to 50 microl/ml.

A novel cytotoxic lignan from Seseli annuum L.

A new tetrahydrofuranoid lignan named seselinone was isolated from the aerial parts of Seseli annuum, together with three known lignans (eudesmin, magnone A and hernone) and prenylated coumarin umbelliprenin. Seselinone and eudesmin showed cytotoxic activity against C6 rat glioma cell cultures.