Combination of Chromatographic Analysis and Chemometric Methods with Bioactivity Evaluation of the Antibacterial Properties of Helichrysum italicum Essential Oil.

Helichrysum italicum (immortelle) essential oil is one of the most popular essential oils worldwide and it has many beneficial properties, including antimicrobial. However, in this plant, the chemical diversity of the essential oil is very pronounced. The aim of this work was to process the GC-MS results of four samples of H. italicum essential oil of Serbian origin by chemometric tools, and evaluate the antimicrobial activity in vitro and in silico. Overall, 47 compounds were identified, the most abundant were γ-curcumene, α-pinene, and ar-curcumene, followed by α-ylangene, neryl acetate, trans-caryophyllene, italicene, α-selinene, limonene, and italidiones. Although the four samples of H. italicum essential oil used in this study were obtained from different producers in Serbia, they belong to the type of essential oil rich in sesquiterpenes (γ-curcumene and ar-curcumene chemotype). In vitro antimicrobial potential showed that five were sensitive among ten strains of tested microorganisms: Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Saccharomyces cerevisiae, and Candida albicans. Therefore, these microorganism models were used further for in silico molecular docking through the mechanism of ATP-ase inhibitory activity. Results showed that among all compounds from H. italicum essential oil, neryl acetate has the highest predicted binding energy. Artificial neural network modeling (ANN) showed that two major compounds γ-curcumene and α-pinene, as well as minor compounds such as trans-ß-ocimene, terpinolene, terpinene-4-ol, isoitalicene, italicene, cis-α-bergamotene, trans-α-bergamotene, italidiones, trans-ß-farnesene, γ-selinene, ß-selinene, α-selinene, and guaiol are responsible for the antimicrobial activity of H. italicum essential oil. The results of this study indicate that H. italicum essential oil samples rich in γ-curcumene, α-pinene, and ar-curcumene cultivated in Serbia (Balkan) have antimicrobial potential both in vitro and in silico. In addition, according to ANN modeling, the proportion of neryl acetate and other compounds detected in these samples has the potential to exhibit antimicrobial activity.

Enhancing Phytoextraction Potential of Brassica napus for Contaminated Dredged Sediment Using Nitrogen Fertilizers and Organic Acids.

Dredged sediment contaminated with heavy metals can be remediated through phytoremediation. The main challenge in phytoremediation is the limited availability of heavy metals for plant uptake, particularly in multi-contaminated soil or sediment. This study aimed to assess the effect of the nitrogen fertilizers (ammonium nitrate (AN), ammonium sulfate (AS), and urea (UR)), organic acids (oxalic (OA) and malic (MA) acids), and their combined addition to sediment on enhancing the bioavailability and phytoremediation efficiency of heavy metals. The sediment dredged from Begej Canal (Serbia) had high levels of Cr, Cd, Cu, and Pb and was used in pot experiments to cultivate energy crop rapeseed (Brassica napus), which is known for its tolerance to heavy metals. The highest accumulation and translocation of Cu, Cd, and Pb were observed in the treatment with AN at a dose of 150 mg N/kg (AN150), in which shoot biomass was also the highest. The application of OA and MA increased heavy metal uptake but resulted in the lowest biomass production. A combination of MA with N fertilizers showed high uptake and accumulation of Cr and Cu.

Brassica Species in Phytoextractions: Real Potentials and Challenges.

The genus Brassica is recognized for including species with phytoaccumulation potential and a large amount of research has been carried out in this area under a variety of conditions, from laboratory experiments to field trials, with spiked or naturally contaminated soils, using one- or multi-element contaminated soil, generating various and sometimes contradictory results with limited practical applications. To date, the actual field potential of Brassica species and the feasibility of a complete phytoextraction process have not been fully evaluated. Therefore, the aim of this study was to summarize the results of the experiments that have been performed with a view to analyzing real potentials and limitations. The reduced biomass and low metal mobility in the soil have been addressed by the development of chemically or biologically assisted phytoremediation technologies, the use of soil amendments, and the application of crop management strategies. Certain issues, such as the fate of harvested biomass or the performance of species in multi-metal-contaminated soils, remain to be solved by future research. Potential improvements to current experimental settings include testing species grown to full maturity, using a greater amount of soil in experiments, conducting more trials under real field conditions, developing improved crop management systems, and optimizing solutions for harvested biomass disposal.

Plant uptake and soil retention of radionuclides and metals in vineyard environments.

In most European countries, each adult citizen drinks on average more than 20 L of wine every year. Three popular wine-growing areas (Aleksandrovac, Topola, and Orahovac) in Serbia were studied in order to investigate the abundance and uptake of elements from vineyard soil to plants. The specific activities of radionuclides (226Ra, 232Th, 40K, 137Cs, and 7Be) were measured in soil, leaves, and grape berries. 226Ra and 232Th were positively correlated with silt and clay and negatively correlated with sand content in soil. Specific activities of natural radionuclides were also negatively correlated with soil pH and CaCO3. Significant correlations of 40K and 137Cs with organic matter in soil were found. Concentrations of fifteen metals (As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Zn, Fe, K, Na, Ca, Mg, and Hg) were also measured in soil samples as well as in grapevine leaves. Analyzed soils were rich in Ni, Cu, Co, Cr, and Cd. High concentrations of Cu were probably caused by long-term use of Cu-based fungicides. Cu was correlated with Fe and organic matter content in soil. Soil-to-plant transfer factors (TF) were calculated to estimate the uptake of radionuclides and metals. Correlations obtained via PCA enable distinction between the sites Aleksandrovac and Topola relative to Orahovac. The first principal component (PC1) accounting for 30.70% of the total variance correlated significantly with soil pH (H2O), contents of CaCO3, Na, Ca, 40K, and 226Ra in soil, as well as with 226Ra, Na, Ca in plants and TFCa. The second principal component (PC2), with total variance of 17.21%, was mainly correlated with variables pertaining to Mg, Co, and Cr in the soil and TFK.

Chemical Characterization of Marrubium vulgare Volatiles from Serbia.

Marrubium vulgare is a cosmopolitan medicinal plant from the Lamiaceae family, which produces structurally highly diverse groups of secondary metabolites. A total of 160 compounds were determined in the volatiles from Serbia during two investigated years (2019 and 2020). The main components were E-caryophyllene, followed by germacrene D, α-humulene and α-copaene. All these compounds are from sesquiterpene hydrocarbons class which was dominant in both investigated years. This variation in volatiles composition could be a consequence of weather conditions, as in the case of other aromatic plants. According to the unrooted cluster tree with 37 samples of Marrubium sp. volatiles from literature and average values from this study, it could be said that there are several chemotypes: E-caryophyllene, ß-bisabolene, α-pinene, ß-farnesene, E-caryophyllene + caryophyllene oxide chemotype, and diverse (unclassified) chemotypes. However, occurring polymorphism could be consequence of adaptation to grow in different environment, especially ecological conditions such as humidity, temperature and altitude, as well as hybridization strongly affected the chemotypes. In addition, this paper aimed to obtain validated models for prediction of retention indices (RIs) of compounds isolated from M. vulgare volatiles. A total of 160 experimentally obtained RIs of volatile compounds was used to build the prediction models. The coefficients of determination were 0.956 and 0.964, demonstrating that these models could be used for predicting RIs, due to low prediction error and high r2.

Marrubium vulgare L.: A Phytochemical and Pharmacological Overview.

Marrubium vulgare is a plant with high bioactive potential. It contains marrubiin, a labdane diterpene that is characteristic for this genus, as well as a complex mixture of phenolic compounds. According to numerous studies, M. vulgare acts as a good antioxidant agent, and due to this, it could potentially be useful in treatments of cancer, diabetes mellitus, and liver diseases. In addition, its anti-inflammatory, wound-healing, antihypertensive, hypolipidemic, and sedative potential are discussed. Apart from that, its antimicrobial activity, especially against Gram+ bacteria, fungi, herpes simplex virus, and parasites such as Toxoplasma gondii, Trichomonas vaginalis, and Plasmodium berghei-berghei was recorded. Additionally, it could be used as a chicken lice repellent, herbicide, and natural insecticide against mosquito larvae and natural molluscicide. In veterinary medicine, M. vulgare can be used as an anthelmintic against the eggs and larvae of bovine strongyles parasites, and as an antibiotic against bovine mastitis caused by resistant bacterial strains. Due to the mentioned benefits, there is a tendency for the cultivation of M. vulgare in order to ensure high-quality raw material, but more firm scientific evidence and well-designed clinical trials are necessary for the well-established use of M. vulgare herb and its preparations.

Is Kragujevac city still a "hot spot" area, twenty years after the bombing?

After NATO bombing of Serbia in 1999, UNEP has identified Kragujevac as one of the four heavily polluted environmental "hot spots". Damaging of industrial and military targets caused the release of substantial amounts of hazardous chemical substances into the environment. This study was conducted in order to access the exposure of residents of Kragujevac city to persistent soil pollutants, twenty years after NATO air campaign. The paper reports the results of measuring radionuclides, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and heavy metals (As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Zn, and Hg) in soil samples collected from two depths (0-15 cm and 15-30 cm) at 30 locations along the riverbank of the Lepenica River. The average specific activities of 226Ra, 232Th, 40K and 137Cs were comparable to average worldwide values; excess lifetime cancer risk (ELCR) from natural radionuclides ranged from 1.1·10-4 to 3.3·10-4. The measured concentrations of As, Co, Cr, Cu, and Ni exceeded the limit values in most of the samples. Non-carcinogenic risk (hazard quotient and hazard index) and carcinogenic risk from heavy metals were assessed. Total hazard index was 0.257 and 2.16 for adults and children, respectively. Sum of measured PAHs ranged from 110 to 1026 µg kg-1. Sum of PCBs exceeded the limit value of 20 µg kg-1 in all samples (it ranged from 48.8 to 196.8 µg kg-1), but it was still below the remediation level. The differences between two layers with respect to all measured variables were not statistically significant.

Spa environments in central Serbia: Geothermal potential, radioactivity, heavy metals and PAHs.

This study aims to estimate geothermal potential, radioactivity levels, and environmental pollution of six most popular spas in Central Serbia (Ovcar, Gornja Trepca, Vrnjacka, Mataruska, Bogutovacka and Sokobanja), as well as to evaluate potential exposure and health risks for living and visiting population. Thermal possibilities of the studied spas showed medium and low geothermal potential with total thermal power of 0.025 MW. Gamma dose rates in air varied from 63 to 178 nSv h-1. Specific activities of natural radionuclides (226Ra, 232Th and 40K) and 137Cs in soil were measured; annual effective doses and excess lifetime cancer risk from radionuclides were calculated. Radon concentration in thermal-mineral waters from the spas ranged between 1.5 and 60.7 Bq L-1 (the highest values were measured in Sokobanja). The annual effective dose from radon due to water ingestion was calculated. The analyzed soils had a clay loam texture. The presence of As, Cr, Cu, Fe, Mn, Ni, Pb, Cd, Zn, and Hg in soil was investigated. The concentrations of As, Cr, Ni, and Hg exceeded the regulatory limits in many samples. Soil samples from Mataruska spa were generally the most contaminated with heavy metals, while the lowest heavy metal concentrations were observed in Sokobanja. Health effects of exposure to heavy metals in soil were estimated by non-carcinogenic risk and carcinogenic risk assessment. Total carcinogenic risk ranged between 6 × 10-4 and 137 × 10-4 for children and between 0.1 × 10-4 and 2.2 × 10-4 for adults. The sum of 16 PAHs analyzed in soil samples varied from 92 to 854 µg kg-1.

Comparison of QuEChERS with Traditional Sample Preparation Methods in the Determination of Multiclass Pesticides in Soil.

Background: The increased use of pesticides leads to permanent pollution of soil, and there is a need for continuous monitoring of these agrochemicals in soil. Objective: Three methods for the simultaneous determination of 12 pesticides belonging to eight chemical groups in soil samples were tested and compared based on analytical parameters. Methods: The quick, easy, cheap, effective, rugged, and safe (QuEChERS); traditional solid-liquid extraction (SLE); and Soxhlet extraction were used for soil sample preparation, while detection and quantification of pesticides were performed using gas chromatography-mass spectrometry (GC-MS). Results: The tested methods featured good sensitivity, and with the exception for carbofuran (Soxhlet method, LOD = 29 µg/kg), for the rest of the pesticides, the studied LODs were less than 12 µg/kg. Except for simazine and carbofuran, LODs obtained by Soxhlet extraction were lower than values obtained by other two methods, whereas QuEChERS gave lower LODs than the traditional SLE method for all compounds except atrazine and acetochlor. The recoveries obtained applying QuEChERS, traditional SLE, and Soxhlet methods for multiple analyses of soil samples fortified at 10, 75, and 200 µg/kg of each pesticide were in the ranges 54-103, 40-91, and 12-92%, respectively. Except for chlorothalonil, the highest recoveries were obtained by the QuEChERS method. Soxhlet was better than traditional SLE method for chlorothalonil, heptachlor, and aldrin; organophosphorus pesticides (fenitrothion and diazinon) and trifluralin, gave similar recoveries for both methods. All three methods were proven to be repeatable, with RSDs lower than 19%. Conclusions: Although all tested methods showed as satisfactory regarding most analytical parameters, QuEChERS method showed much better results in terms of confidence, indicating that traditional SLE and Soxhlet extraction still need improvements for determination of multiclass pesticides in soil samples.

Environmental risk assessment of radioactivity and heavy metals in soil of Toplica region, South Serbia.

Activity levels of natural and artificial radionuclides and content of ten heavy metals (As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Zn and Hg) were investigated in 41 soil samples collected from Toplica region located in the south part of Serbia. Radioactivity was determined by gamma spectrometry using HPGe detector. The obtained mean activity concentrations ± standard deviations of radionuclides 226Ra, 232Th, 40K and 137Cs were 29.9 ± 9.4, 36.6 ± 11.5, 492 ± 181 and 13.4 ± 18.7 Bq kg-1, respectively. According to Shapiro-Wilk normality test, activity concentrations of 226Ra and 232Th were consistent with normal distribution. External exposure from radioactivity was estimated through dose and radiation risk assessments. Concentrations of heavy metals were measured by using ICP-OES, and their health risks were then determined. Enrichment by heavy metals and pollution level in soils were evaluated using the enrichment factor, the geoaccumulation index (Igeo), pollution index and pollution load index. Based on GIS approach, the spatial distribution maps of radionuclides and heavy metal contents were made. Spearman correlation coefficient was used for correlation analysis between radionuclide activity concentrations and heavy metal contents.

Persistent organic pollutants, heavy metals and radioactivity in the urban soil of Pristina City, Kosovo and Metohija.

Polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), heavy metals content and radioactivity levels were measured in 27 soil samples collected from Pristina, the capital of Kosovo and Metohija. The sixteen PAHs, twelve OCPs and six PCBs congeners were determined by gas chromatography system with mass spectrometry detection. Although the use of PCBs and OCPs was prohibited decades ago residues of those compounds still existed in measurable concentrations in soils of Pristina. PAHs were also present in analyzed samples but their mean concentration was significantly lower than mean concentrations of PAHs previously reported in urban areas in the world. The concentrations of heavy metals (As, Cd, Co, Cr, Cu, Mn, Ni, Pb and Zn) were determined by the EDTA extraction protocols, along with their extractable concentrations. The activity concentrations of natural radionuclides (226Ra, 232Th and 40K) were determined by gamma spectrometry method. The Shapiro-Wilk normality test found that activity concentrations of natural radionuclides were normally distributed. Radiological risk was estimated through the annual effective dose, gonadal dose equivalent, excess lifetime cancer risk, radium equivalent activity, external and internal hazard indexes. Spearman correlation coefficient was used for analysis of correlations between physicochemical properties, heavy metal contents and radionuclide activity concentrations. Strong positive correlation between 226Ra and 232Th was found, as well as among pairs of As-Cd and Co-Mn. Very strong positive correlation (0.838) at the 0.01 significance level was noted for Pb-Zn pair. Strong correlations indicate common occurrence of these elements in the nature, as well as geogenic association.

Transfer factors of natural radionuclides and (137)Cs from soil to plants used in traditional medicine in central Serbia.

Transfer factors of natural radionuclides and (137)Cs from soil to plants used in traditional medicine were determined. The transfer factors (TF) were calculated as Bq kg(-1) of dry plant per Bq kg(-1) of dry soil. Mass activity concentrations of (226)Ra, (232)Th, (40)K and (137)Cs in soil and plant samples were measured with high purity germanium detector (HPGe). The concentrations of As, Co, Cr, Cu, Mn, Ni, Pb and Zn were determined, as well as the cation exchange capacity (CEC) and the content of exchangeable cations (Ca, Mg, K, Na). Wide ranges of values were obtained for all the metals, especially for Cr and Ni. The Absalom model was used for determination of the amount of (137)Cs transferred from soil to plant based on soil characteristics such as pH, exchangeable potassium, humus and clay contents. The estimated transfer factors were in the range from 0.011 to 0.307 with an arithmetic mean of 0.071, median of 0.050, geometric mean of 0.053 and geometric standard deviation (GSD) of 2.08. This value agreed well with that calculated from the measurements of 0.069, geometric mean 0.040 and GSD 3.19. Correlations between radionuclides, metals, physicochemical properties and transfer factors were determined by Spearman correlation coefficient. There was a strong positive correlation between (137)Cs transfer factor and the ratio of transfer factor for K and (137)Cs. Principal Component Analysis (PCA) was performed in order to identify some pattern of data.

Correlation between radioactivity levels and heavy metal content in the soils of the North Kosovska Mitrovica environment.

This paper reports the results of radioactivity concentrations and heavy metal content in soil samples collected from non-agricultural areas in the municipality and vicinity of North Kosovska Mitrovica, formerly the most important mining area in Europe. The estimated average activity concentrations of (226)Ra, (232)Th, (40)K and (137)Cs are 40.6 ± 19, 48 ± 25.4, 743.2 ± 200.5 and 81 ± 119 Bq kg(-1), respectively. The estimated average absorbed dose rate in the air and the annual effective dose are 78.7 nGy h(-1) and 96.6 µSv, respectively. The radium equivalent activity and external hazard index were also calculated. High contents of Pb, Zn, Cu, Cd, As and Ni were found in the analyzed soil samples, thus indicating pollution of the environment. Most metals have a wide range of values spanning 2 to 3 orders of magnitude, which is particularly evident for Pb and Zn. Correlations between the activity concentrations of the radionuclides, the heavy metal content in soil and the basic soil properties were determined by means of the Pearson linear coefficient. Strong positive correlations between the naturally occurring radionuclides, and also among Pb, Zn, Cu and Cd were found.